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Sample records for hahkhallituse botrytis cinerea

  1. Electron Microscopy of Botrytis cinerea Conidia

    PubMed Central

    Buckley, Patricia M.; Sjaholm, Virginia E.; Sommer, N. F.

    1966-01-01

    Buckley, Patricia M. (University of California, Davis), Virginia E. Sjaholm, and N. F. Sommer. Electron microscopy of Botrytis cinerea conidia. J. Bacteriol. 91:2037–2044. 1966.—Sections of germinating and nongerminating Botrytis cinerea conidia were examined with an electron microscope. Uranyl acetate or lead citrate provided contrast between membranes and cytoplasm. Membrane-bounded, dense inclusions previously unreported in dormant spores were termed “storage bodies.” Whorled structures, spherules, granules, and membrane loops were seen within these inclusions. The various forms assumed by the enclosed materials closely resemble phospholipid inclusions described for other cells. It is suggested that the inclusions provide material for the assembly of membranous organelles during germination. Utilization of the stored material apparently results in extensive vacuolization in advanced germinants. Images PMID:5949251

  2. Three new metabolites from Botrytis cinerea.

    PubMed

    Wang, Tian-Shan; Zhou, Jin-Yan; Tan, Hong

    2008-01-01

    Three new metabolites, gamma-abscisolactone (1), botrytisic acids A (3) and B (4) were isolated from the fermentation broth of Botrytis cinerea TB-3-H8. Their structures were elucidated on the basis of MS, IR, UV, and NMR spectroscopic data. Compound 2 was isolated from natural resource for the first time. The structure of 1 was further confirmed by single-crystal X-ray diffraction (CCDC-265897). PMID:19003608

  3. Adhesion of germlings of Botrytis cinerea.

    PubMed Central

    Doss, R P; Potter, S W; Soeldner, A H; Christian, J K; Fukunaga, L E

    1995-01-01

    Adhesion of conidia and germlings of the facultative plant parasite Botrytis cinerea occurs in two distinct stages. The first stage, which occurs immediately upon hydration of conidia and is characterized by relatively weak adhesive forces, appears to involve hydrophobic interactions (R. P. Doss, S. W. Potter, G. A. Chastagner, and J. K. Christian, Appl. Environ. Microbiol. 59:1786-1791, 1993). The second stage of adhesion, delayed adhesion, occurs after viable conidia have been incubated for several hours under conditions that promote germination. At this time, the germlings attach strongly to either hydrophobic or hydrophilic substrata. Delayed adhesion involves secretion of an ensheating film that remains attached to the substratum upon physical removal of the germlings. This fungal sheath, which can be visualized by using interference-contrast light microscopy, scanning electron microscopy, or atomic force microscopy, is 25 to 60 nm thick in the region immediately adjacent to the germ tubes. Germlings are resistant to removal by boiling or by treatment with a number of hydrolytic enzymes, 2.0 M periodic acid, or 1.0 M sulfuric acid. They are readily removed by brief exposure to 1.25 N NaOH. A base-soluble material that adheres to culture flask walls in short-term liquid cultures of B. cinerea is composed of glucose (about 30%), galactosamine (about 3%), and protein (30 to 44%). PMID:7887606

  4. Fungicide resistance profiles for 13 Botrytis cinerea isolated from strawberry in southeastern Louisiana

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BACKGROUND: Fungicidal sprays have been widely used for disease control of gray mold caused by Botrytis cinerea. In recent years strawberry growers in southeastern Louisiana reported a failure of their fungicide spray programs to control Botrytis fruit rot. Botrytis cinerea has become resistant ...

  5. Isolate Dependency of Brassica rapa Resistance QTLs to Botrytis cinerea.

    PubMed

    Zhang, Wei; Kwon, Soon-Tae; Chen, Fang; Kliebenstein, Daniel J

    2016-01-01

    Generalist necrotrophic pathogens including Botrytis cinerea cause significant yield and financial losses on Brassica crops. However, there is little knowledge about the mechanisms underlying the complex interactions encoded by both host and pathogen genomes in this interaction. This potentially includes multiple layers of plant defense and pathogen virulence mechanisms that could complicate in breeding broad spectrum resistance within Brassica species. Glucosinolates (GSLs) are a diverse group of defense metabolites that play a key role in interaction between Brassica and biotic attackers. In this study, we utilized a collection of diverse B. cinerea isolates to investigate resistance within the Brassica rapa R500 × IMB211 recombinant inbred line population. We tested variation on lesion development and glucosinolate accumulation in parental lines and all population lines. We then mapped quantitative trait loci (QTL) for both resistances to B. cinerea and defense metabolites in this population. Phenotypic analysis and QTL mapping demonstrate that the genetic basis of resistance to B. cinerea in B. rapa is isolate specific and polygenic with transgressive segregation that both parents contribute resistance alleles. QTLs controlling defensive GSLs are highly dependent on pathogen infection. An overlap of two QTLs identified between resistance to B. cinerea and defense metabolites also showed isolate specific effects. This work suggests that directly searching for resistance loci may not be the best approach at improving resistance in B. rapa to necrotrophic pathogen. PMID:26925079

  6. Isolate Dependency of Brassica rapa Resistance QTLs to Botrytis cinerea

    PubMed Central

    Zhang, Wei; Kwon, Soon-Tae; Chen, Fang; Kliebenstein, Daniel J.

    2016-01-01

    Generalist necrotrophic pathogens including Botrytis cinerea cause significant yield and financial losses on Brassica crops. However, there is little knowledge about the mechanisms underlying the complex interactions encoded by both host and pathogen genomes in this interaction. This potentially includes multiple layers of plant defense and pathogen virulence mechanisms that could complicate in breeding broad spectrum resistance within Brassica species. Glucosinolates (GSLs) are a diverse group of defense metabolites that play a key role in interaction between Brassica and biotic attackers. In this study, we utilized a collection of diverse B. cinerea isolates to investigate resistance within the Brassica rapa R500 × IMB211 recombinant inbred line population. We tested variation on lesion development and glucosinolate accumulation in parental lines and all population lines. We then mapped quantitative trait loci (QTL) for both resistances to B. cinerea and defense metabolites in this population. Phenotypic analysis and QTL mapping demonstrate that the genetic basis of resistance to B. cinerea in B. rapa is isolate specific and polygenic with transgressive segregation that both parents contribute resistance alleles. QTLs controlling defensive GSLs are highly dependent on pathogen infection. An overlap of two QTLs identified between resistance to B. cinerea and defense metabolites also showed isolate specific effects. This work suggests that directly searching for resistance loci may not be the best approach at improving resistance in B. rapa to necrotrophic pathogen. PMID:26925079

  7. Grape Berry Colonization and Biological Control of Botrytis cinerea by Indigenous Vineyard Yeasts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis bunch rot, caused by Botrytis cinerea, is the most important disease of grape berries, especially during transportation and storage. Biological control is a potential means of postharvest management of Botrytis bunch rot. The study was aimed at testing the hypothesis that antagonistic yeast...

  8. Adhesion of Nongerminated Botrytis cinerea Conidia to Several Substrata †

    PubMed Central

    Doss, Robert P.; Potter, Sandra W.; Chastagner, Gary A; Christian, James K.

    1993-01-01

    Conidia of the plant pathogenic fungus Botrytis cinerea adhered to tomato cuticle and to certain other substrata immediately upon hydration. This immediate adhesion occurred with both living and nonliving conidia. Adhesion was not consistently influenced by several lectins, sugars, or salts or by protease treatment, but it was strongly inhibited by ionic or nonionic detergents. With glass and oxidized polyethylene, substrata whose surface hydrophobicities could be conveniently varied, there was a direct relationship between water contact angle and percent adhesion. Immediate adhesion did not involve specific conidial attachment structures, although the surfaces of attached conidia were altered by contact with a substratum. Freshly harvested conidia were very hydrophobic, with more than 97% partitioning into the organic layer when subjected to a phase distribution test. Percent adhesion of germinated conidia was larger than that of nongerminated conidia. Evidence suggests that immediate adhesion of conidia of B. cinerea depends, at least in part, on hydrophobic interactions between the conidia and substratum. Images PMID:16348954

  9. Action Spectrum for Photosporogenesis in Botrytis cinerea Pers. ex Fr

    PubMed Central

    Honda, Yuichi; Yunoki, Toshifumi

    1978-01-01

    An action spectrum for photoinduced conidium formation in the fungus Botrytis cinerea Pers. ex Fr. was determined by exposing colonies to monochromatic radiation obtained from a diffraction grating monochromator. Wavelengths longer than 355 nm were ineffective even after exposures of 8 hours at intensities of 4,068 to 8,276 erg per cm2 per second. Colonies were exposed at 22.5 C from 226 to 355 nm at about 5 nm intervals. Three prominent peaks of effectiveness occurred at about 231, 268, and 283 nm, and there was also a minor peak at about 303 nm. The most effective wavelength for inducing sporulation was 231 nm; it was 25% more effective than the second most effective wavelength (283 nm). PMID:16660370

  10. Role of temperature and free moisture in onion flower blight. [Botrytis squamosa; Botrytis cinerea; and Botrytis allii

    SciTech Connect

    Ramsey, G.R.; Lorbeer, J.W.

    1986-06-01

    The cardinal temperatures at which onion umbels were blighted (after inoculation when two-thirds of the florets were open) with Botrytis squamosa, B. cinerea, and B. allii (isolated from blighted onion florets) were near 9, 21, and 27 C for B. squamosa, near 12, 21, and 30 C for B. cinerea, and near 9, 24, and 30 C for B. allii. The cardinal temperatures for mycelial growth (potato-dextrose agar) of B. squamosa, B. cinerea, and B. allii were near 5, 22, and 30 C for each fungus. The cardinal temperatures for conidial germination (on purified water agar) were near 6, 15, and 30 C for B. squamosa; 3, 18, and 33 C for B. cinerea; and 6, 24, and 33 C for B. allii. When the duration of free moisture on umbels after inoculation with the three pathogens was increased from 0 to 96 hr. the percentages of unopened florets, open florets, and immature seed capsules blighted at 21 C were increased significantly. Free moisture durations of 12-24, 6-12, and 6-12 hr were necessary for blighting of unopen florets, open florets, and immature seed capsules, respectively, by each pathogen at 21 C. A positive correlation between the amount of July rainfall and the natural incidence of onion flower blight was observed in Orange County, New York, from 1976 to 1981. 10 references, 2 figures, 1 table.

  11. First Report of Botrytis cinerea as a Postharvest Pathogen of Blueberry in Korea

    PubMed Central

    Cheon, Mi-Geon; Choi, Okhee; Kim, Jinwoo

    2011-01-01

    Gray mold of blueberry caused by Botrytis sp. is reported for the first time in Korea. A detailed description of the fungus is given, along with its rDNA internal transcribed spacer sequence. The fungus was identified as Botrytis cinerea based on mycological characteristics and molecular data. PMID:22783073

  12. Heat-induced oxidative injury contributes to inhibition of Botrytis cinerea spore germination and growth

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The inhibitory effect of a heat treatment (HT) on Botrytis cinerea, a major postharvest fungal pathogen, and the possible mode of action were investigated. Spore germination and germ tube elongation of B. cinerea were both increasingly and significantly inhibited by a HT (43 degrees C) for 10, 20 o...

  13. A double-stranded RNA mycovirus confers hypovirulence-associated traits to Botrytis cinerea.

    PubMed

    Castro, Miguel; Kramer, Kurt; Valdivia, Lorena; Ortiz, Sylvia; Castillo, Antonio

    2003-11-01

    Botrytis cinerea CCg425 contains a 33-nm isometric mycovirus whose genome is a 6.8-kb double-stranded RNA (dsRNA) molecule. Virulence bioassays, performed by direct plug mycelial inoculation on bean plant leaves, showed that B. cinerea CCg425 displays less fungal aggressivity than B. cinerea CKg54, a virulent fungal strain that is not infected by dsRNA mycoviruses. B. cinerea CCg425 also showed lower laccase activity and conidiation rate than B. cinerea CKg54. Furthermore, infection of B. cinerea CKg54 with viral particles purified from B. cinerea CCg425 resulted in diminished virulence of the infected fungus. Collectively, our results indicate that mycovirus infection confers hypovirulence to the fungal host. PMID:14612241

  14. Novel hypovirulence-associated RNA mycovirus in the plant pathogenic fungus botrytis cinerea: molecular and biological characterization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis cinerea is a pathogenic fungus causing gray mold disease on numerous economically important crops and ornamental plants. This study was conducted to characterize the biological and molecular features of a novel RNA mycovirus, Botrytis cinerea RNA virus 1 (BcRV1), in the hypovirulent strain ...

  15. Effect of calcium on cell-wall degrading enzymes of Botrytis cinerea.

    PubMed

    Sasanuma, Izumi; Suzuki, Takuya

    2016-09-01

    Effective anti-Botrytis strategies leading to reduce pesticides on strawberries are examined to provide the protection that is harmless to humans, higher animals and plants. Calcium treatments significantly inhibited the spore germination and mycelial growth of B. cinerea. The intracellular polygalacturonase and CMCase showed low activities in B. cinerea cultivated by medium containing calcium. On the other hand, calcium-stimulated β-glucosidases production occurred. Our findings suggest that the calcium treatments keep CMCase activity low and cause low activities of cell-wall degrading enzymes of B. cinerea in the late stage of growth. PMID:26998660

  16. Development of a selective medium for the determination of the spore concentrations of Botrytis cinerea in the air.

    PubMed

    Gielen, S; Aerts, R; Seels, B

    2003-01-01

    Gray mold, caused by Botrytis cinerea, is an important disease that causes world-wide extensive damage to a wide range of economically important crops. When it is necessary to determine the spore concentration of Botrytis cinerea in a certain area, it is important to develop a method that can capture the spores of Botrytis cinerea and that can identify them. For the identification and enumeration of the plant pathogen Botrytis cinerea in the environment the easiest method available for the moment is the use of a selective medium. Several selective media for the isolation of Botrytis spp. have been developed by other research groups. All these media contain fungicides that are usually non-toxic towards Botrytis species and tannic acid, which is oxidized to produce a brown pigment that visualises the growth of Botrytis cinerea on the selective media. It seemed that different isolates of Botrytis cinerea that are found in nature have different sensitivities towards the different fungicide concentrations that are used in the selective media. Making the "optimal" selective media for Botrytis cinerea, we have to take in consideration that so many as possible Botrytis cinerea isolates must be able to germinate and grow on this selective medium and that the contamination of other micro-organisms on the selective medium must be minimized. Before the final composition of our selective medium for Botrytis cinerea, different combinations of fungicide concentrations were tried out of the following three fungicides: Rubigan, maneb and PCNB (pentachloronitrobenzene). All these selective media with different fungicides concentrations were tested out for spore germination and mycelium growth of Botrytis cinerea. Because it was obvious that the percentage Botrytis cinerea that germinated on PDA (Potato Dextrose Agar) was higher than on the selective medium a few experiments were executed in which the percentage of spore germination on PDA was compared with the percentage of spore

  17. First report of pyrimethanil resistance in Botrytis cinerea from stored apples in Pennsylvania

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis cinerea Pers.: Fr. (teleomorph Botryotinia fuckeliana [de Bary] Whetzel) causes gray mold on apple fruit which is an economically significant disease. A survey of commercial packinghouses in Washington State revealed that gray mold accounted for 28% of the decayed apples in storage. Fungici...

  18. Practical resistance to fenhexamid Botrytis cinerea isolates from grapevines in New York

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fenhexamid is a fungicide used to control Botrytis cinerea on grapes worldwide. Resistance appears to be of a quantitative rather than qualitative nature, with minimum EC50 values that define a resistant phenotype proposed as exceeding 0.1 mg/L by some workers and 0.4 mg/L by others. However, little...

  19. Sodium hypochlorite: A promising agent for reducing Botrytis cinerea infection on rose flowers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis cinerea is a fungal pathogen that greatly reduces the postharvest quality of rose flowers. We determined the potential of sodium hypochlorite (NaOCl), an oxidizer with antimicrobial activity, to reduce the incidence of disease on flowers. A postharvest dip in 200 µL L-1 NaOCl for 10 s at ...

  20. Comparative analysis of techniques for detection of quiescent Botrytis cinerea in grapes by quantitative PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Quantitative PCR (qPCR) can be used to detect and monitor pathogen colonization, but early attempts to apply the technology to quiescent Botrytis cinerea infections of grape berries identified some specific limitations. In this study, four DNA extraction methods, two tissue-grinding methods, two gra...

  1. Decay resistance to Botrytis cinerea and quality characteristics during storage of raspberry genotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Raspberries are a delicate, high value crop with an extremely short shelf life exacerbated by postharvest decay caused by Botrytis cinerea Pers. European red raspberry (Rubus idaeus L.) is the most widely grown. Yellow (R. idaeus L.), black (R. occidentalis L.) and purple raspberries (R. ×neglectus ...

  2. Selection, fitness, and control of grape isolates of Botrytis cinerea variably sensitive to fenhexamid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Of 683 Botrytis cinerea isolates collected from a fungicide-trial vineyard, 31 were classified as putatively resistant to fenhexamid (EC50 =0.1 ug/ml). For the resistant isolates that survived and sporulated in culture, colony expansion and conidial germination frequency was significantly reduced re...

  3. Botrytis californica, a new cryptic species in the B. cinerea species complex causing gray mold in blueberries and table grapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis cinerea consists of two cryptic species, referred to as Group I and Group II based on Bc-hch gene RFLP haplotyping, and Group I has been described as a new cryptic species B. pseudocinerea. During a survey for Botrytis spp. causing gray mold in blueberries and table grapes in the Central Va...

  4. Effect of cucurbitacins on mRNA coding for laccase in Botrytis cinerea.

    PubMed

    Gonen, L; Viterbo, A; Cantone, F; Staples, R C; Mayer, A M

    1996-05-01

    The effect of cucurbitacin and of Ecballium extract on the formation of mRNA coding for laccase was examined in cultures of Botrytis cinerea grown with inducers of laccase formation, in the presence or absence of the inhibitory compounds. RNA was isolated from the cultures and probed with specific DNA probes for laccase. As an internal control, the RNA was probed for Botrytis beta-tubulin mRNA. From an analysis of the results it is clear that cucurbitacin I and Ecballium extract specifically repress the amount of mRNA coding for laccase. This could account for the previously observed repression of laccase formation by cucurbitacins. PMID:8688171

  5. Interkingdom Gene Transfer May Contribute to the Evolution of Phytopathogenicity in Botrytis Cinerea

    PubMed Central

    Zhu, Bo; Zhou, Qing; Xie, Guanlin; Zhang, Guoqing; Zhang, Xiaowei; Wang, Yanli; Sun, Gunchang; Li, Bin; Jin, Gulei

    2012-01-01

    The ascomycete Botrytis cinerea is a phytopathogenic fungus infecting and causing significant yield losses in a number of crops. The genome of B. cinerea has been fully sequenced while the importance of horizontal gene transfer (HGT) to extend the host range in plant pathogenic fungi has been recently appreciated. However, recent data confirm that the B. cinerea fungus shares conserved virulence factors with other fungal plant pathogens with narrow host range. Therefore, interkingdom HGT may contribute to the evolution of phytopathogenicity in B. cinerea. In this study, a stringent genome comparison pipeline was used to identify potential genes that have been obtained by B. cinerea but not by other fungi through interkingdom HGT. This search led to the identification of four genes: a UDP-glucosyltransferase (UGT), a lipoprotein and two alpha/beta hydrolase fold proteins. Phylogenetic analysis of the four genes suggests that B. cinerea acquired UGT from plants and the other 3 genes from bacteria. Based on the known gene functions and literature searching, a correlation between gene acquision and the evolution of pathogenicity in B. cinerea can be postulated. PMID:22346340

  6. The effect of harpin on shelf life of peppers inoculated with Botrytis cinerea.

    PubMed

    Tezcan, Himmet; Akbudak, Nuray; Akbudak, Bulent

    2013-12-01

    The preservation methods as an alternative to chemical control to prevent postharvest quality losses of peppers were examined. The efficacy of harpin treatments on peppers (Capsicum annuum L. cvs. 'Demre', 'Yalova Charleston' and 'Sari Sivri') was tested in the same conditions in two different years. Peppers grown in greenhouse were applied with four treatments consisting of harpin, Botrytis cinerea, harpin+B. cinerea and control. The harpin in B. cinerea treatments reduced the percentage of rotten fruit in cv. 'Demre' from 42.68% to 22.85%, in cv. 'Yalova Charleston' from 60.87% to 26.59% and in cv. 'Sari Sivri' from 32.83% to 12.82%. The harpin and harpin+B. cinerea peppers had a better overall appearance at the end of shelf-life. Changes in percentage of red fruit and fruit color at the end of shelf life proceeded more slowly in the harpin treated fruit. The treatments of harpin gave the best results in all three cultivars. Moreover, the values obtained from fruits subjected to harpin+B. cinerea were better than those of the fruits picked from the plants only subjected to B. cinerea. In the trials, harpin slowed down the changes leading to quality loss in fruits, in all cultivars. Thus, the positive effect of harpin was revealed more clearly especially in the fruits picked from the inoculated plants. PMID:24426019

  7. Interactions involving ozone, Botrytis cinerea, and B. squamosa on onion leaves

    SciTech Connect

    Rist, D.L.

    1983-01-01

    Interactions involving Botrytis cinerea Pers., B. squamosa Walker, and ozone on onion (alium cepae L.) were investigated. Initially, threshold dosages of ozone required to predispose onion leaves to greater infection by B. cinerea and B. squamosa were determined under controlled conditions in an ozone-exposure chamber. Subsequent experiments supported the hypothesis that nutrients leaking out of ozone-injured cells stimulated lesion production by B. cinerea. The electrical conductivity of, and carbohydrate concentration in, dew collected from leaves of onion plants which had been exposed to ozone were greater than the electrical conductivity of, and carbohydrate concentration in, dew collected from leaves of other, non-exposed onion plants. When conidia of B. cinerea were suspended in dew collected from leaves of plants which had been exposed to ozone and the resulting suspension atomized onto leaves of non-exposed plants, more lesions were induced than on leaves of other non-exposed plants inoculated with conidia suspended in dew collected from plants which had not been exposed to ozone. EDU protected onion leaves from ozone-induced predisposition to these fungi under controlled conditions. Experiments designed to detect interaction between B. cinerea and B. squamosa in onion leaf blighting indicated that such interaction did not occur. Leaves were blighted rapidly when inoculated with B. squamosa whether B. cinerea was present or absent.

  8. Transformation of Botrytis cinerea by direct hyphal blasting or by wound-mediated transformation of sclerotia

    PubMed Central

    2011-01-01

    Background Botrytis cinerea is a haploid necrotrophic ascomycete which is responsible for 'grey mold' disease in more than 200 plant species. Broad molecular research has been conducted on this pathogen in recent years, resulting in the sequencing of two strains, which has generated a wealth of information toward developing additional tools for molecular transcriptome, proteome and secretome investigations. Nonetheless, transformation protocols have remained a significant bottleneck for this pathogen, hindering functional analysis research in many labs. Results In this study, we tested three different transformation methods for B. cinerea: electroporation, air-pressure-mediated and sclerotium-mediated transformation. We demonstrate successful transformation with three different DNA constructs using both air-pressure- and sclerotium-mediated transformation. Conclusions These transformation methods, which are fast, simple and reproducible, can expedite functional gene analysis of B. cinerea. PMID:22188865

  9. Metabolic activities of five botryticides against Botrytis cinerea examined using the Biolog FF MicroPlate.

    PubMed

    Wang, Hancheng; Wang, Jin; Li, Licui; Hsiang, Tom; Wang, Maosheng; Shang, Shenghua; Yu, Zhihe

    2016-01-01

    Tobacco grey mold caused by Botrytis cinerea is an important fungal disease worldwide. Boscalid, carbendazim, iprodione, pyrimethanil and propiconazole are representative botryticides for grey mold management. This research investigated the sensitivities of B. cinerea from tobacco to these chemicals using the Biolog FF Microplate. All five chemicals showed inhibitory activity, with average EC50 values of 0.94, 0.05, 0.50, 0.61 and 0.31 μg ml(-1), respectively. B. cinerea metabolized 96.8% of tested carbon sources, including 29 effectively and 33 moderately, but the metabolic fingerprints differed under pressures imposed by these botryticides. For boscalid, B. cinerea was unable to metabolize many substrates related to tricarboxylic acid cycle. For carbendazim, carbon sources related to glycolysis were not metabolized. For iprodione, use of most carbon substrates was weakly inhibited, and the metabolic profile was similar to that of the control. For propiconazole, no carbon substrates were metabolized and the physiological and biochemical functions of the pathogen were totally inhibited. These findings provide useful information on metabolic activities of these botryticides, and may lead to future applications of the Biolog FF Microplate for examining metabolic effects of other fungicides on other fungi, as well as providing a metabolic fingerprint of B. cinerea that could be useful for identification. PMID:27491536

  10. Large-Scale Transcriptome Analysis of Cucumber and Botrytis cinerea during Infection

    PubMed Central

    Liu, Tingting; Zhu, Jing; Wang, Jingqi; He, Xiaoqing; Jin, Yi

    2015-01-01

    Cucumber gray mold caused by Botrytis cinerea is considered one of the most serious cucumber diseases. With the advent of Hi-seq technology, it is possible to study the plant–pathogen interaction at the transcriptome level. To the best of our knowledge, this is the first application of RNA-seq to identify cucumber and B. cinerea differentially expressed genes (DEGs) before and after the plant–pathogen interaction. In total, 248,908,688 raw reads were generated; after removing low-quality reads and those containing adapter and poly-N, 238,341,648 clean reads remained to map the reference genome. There were 3,512 cucumber DEGs and 1,735 B. cinerea DEGs. GO enrichment and KEGG enrichment analysis were performed on these DEGs to study the interaction between cucumber and B. cinerea. To verify the reliability and accuracy of our transcriptome data, 5 cucumber DEGs and 5 B. cinerea DEGs were chosen for RT-PCR verification. This is the first systematic transcriptome analysis of components related to the B. cinerea–cucumber interaction. Functional genes and putative pathways identified herein will increase our understanding of the mechanism of the pathogen–host interaction. PMID:26536465

  11. Metabolic activities of five botryticides against Botrytis cinerea examined using the Biolog FF MicroPlate

    PubMed Central

    Wang, Hancheng; Wang, Jin; Li, Licui; Hsiang, Tom; Wang, Maosheng; Shang, Shenghua; Yu, Zhihe

    2016-01-01

    Tobacco grey mold caused by Botrytis cinerea is an important fungal disease worldwide. Boscalid, carbendazim, iprodione, pyrimethanil and propiconazole are representative botryticides for grey mold management. This research investigated the sensitivities of B. cinerea from tobacco to these chemicals using the Biolog FF Microplate. All five chemicals showed inhibitory activity, with average EC50 values of 0.94, 0.05, 0.50, 0.61 and 0.31 μg ml−1, respectively. B. cinerea metabolized 96.8% of tested carbon sources, including 29 effectively and 33 moderately, but the metabolic fingerprints differed under pressures imposed by these botryticides. For boscalid, B. cinerea was unable to metabolize many substrates related to tricarboxylic acid cycle. For carbendazim, carbon sources related to glycolysis were not metabolized. For iprodione, use of most carbon substrates was weakly inhibited, and the metabolic profile was similar to that of the control. For propiconazole, no carbon substrates were metabolized and the physiological and biochemical functions of the pathogen were totally inhibited. These findings provide useful information on metabolic activities of these botryticides, and may lead to future applications of the Biolog FF Microplate for examining metabolic effects of other fungicides on other fungi, as well as providing a metabolic fingerprint of B. cinerea that could be useful for identification. PMID:27491536

  12. Attachment Capability of Antagonistic Yeast Rhodotorula glutinis to Botrytis cinerea Contributes to Biocontrol Efficacy

    PubMed Central

    Li, Boqiang; Peng, Huaimin; Tian, Shiping

    2016-01-01

    Rhodotorula glutinis as an antagonism show good biocontrol performance against various post-harvest diseases in fruits. In the present study, strong attachment capability of R. glutinis to spores and hyphae of Botrytis cinerea was observed. Further analysis showed that certain protein components on the yeast cell surface played critical role during the interaction between R. glutinis and B. cinerea. The components mainly distributed at the poles of yeast cells and might contain glycosylation modification, as tunicamycin treated yeast cells lost attachment capability to B. cinerea. To investigate contributions of attachment capability of R. glutinis to its biocontrol efficacy, yeast cells were mutagenized with 3% methane-sulfonic acid ethyl ester (EMS), and a mutant CE4 with stable non-attaching phenotype was obtained. No significant difference was found on colony, cell morphology, reproductive ability, and capsule formation between the mutant and wild-type. However, there was a distinct difference in India ink positive staining patterns between the two strains. Moreover, wild-type strain of R. glutinis showed better performance on inhibiting spore germination and mycelial growth of B. cinerea than CE4 strain when yeast cells and B. cinerea were co-cultured in vitro. In biocontrol assay, both wild-type and CE4 strains showed significant biocontrol efficacy against gray mold caused by B. cinerea in apple fruit, whereas, control effect of CE4 strain was lower than that of wild-type. Our findings provided new evidences that attachment capability of R. glutinis to B. cinerea contributed to its biocontrol efficacy. PMID:27199931

  13. Attachment Capability of Antagonistic Yeast Rhodotorula glutinis to Botrytis cinerea Contributes to Biocontrol Efficacy.

    PubMed

    Li, Boqiang; Peng, Huaimin; Tian, Shiping

    2016-01-01

    Rhodotorula glutinis as an antagonism show good biocontrol performance against various post-harvest diseases in fruits. In the present study, strong attachment capability of R. glutinis to spores and hyphae of Botrytis cinerea was observed. Further analysis showed that certain protein components on the yeast cell surface played critical role during the interaction between R. glutinis and B. cinerea. The components mainly distributed at the poles of yeast cells and might contain glycosylation modification, as tunicamycin treated yeast cells lost attachment capability to B. cinerea. To investigate contributions of attachment capability of R. glutinis to its biocontrol efficacy, yeast cells were mutagenized with 3% methane-sulfonic acid ethyl ester (EMS), and a mutant CE4 with stable non-attaching phenotype was obtained. No significant difference was found on colony, cell morphology, reproductive ability, and capsule formation between the mutant and wild-type. However, there was a distinct difference in India ink positive staining patterns between the two strains. Moreover, wild-type strain of R. glutinis showed better performance on inhibiting spore germination and mycelial growth of B. cinerea than CE4 strain when yeast cells and B. cinerea were co-cultured in vitro. In biocontrol assay, both wild-type and CE4 strains showed significant biocontrol efficacy against gray mold caused by B. cinerea in apple fruit, whereas, control effect of CE4 strain was lower than that of wild-type. Our findings provided new evidences that attachment capability of R. glutinis to B. cinerea contributed to its biocontrol efficacy. PMID:27199931

  14. Effects of ozone treatment on Botrytis cinerea and Sclerotinia sclerotiorum in relation to horticultural product quality.

    PubMed

    Sharpe, Deana; Fan, Lihua; McRae, Ken; Walker, Brad; MacKay, Ron; Doucette, Craig

    2009-08-01

    Botrytis cinerea and Sclerotinia sclerotiorum are fungal pathogens that cause the decay of many fruits and vegetables. Ozone may be used as an antimicrobial agent to control the decay. The effect of gaseous ozone on spore viability of B. cinerea and mycelial growth of B. cinerea and S. sclerotiorum were investigated. Spore viability of B. cinerea was reduced by over 99.5% (P < 0.01) and height of the aerial mycelium was reduced from 4.7 mm in the control to less than 1 mm after exposure to 450 or 600 ppb ozone for 48 h at 20 degrees C. Sporulation of B. cinerea was also substantially inhibited by ozone treatments. However, ozone had no significant effect on mycelial growth of S. sclerotiorum in vitro. Decay and quality parameters including color, chlorophyll fluorescence (CF), and ozone injury were further assessed for various horticultural commodities (apple, grape, highbush blueberry, and carrot) treated with 450 ppb of ozone for 48 h at 20 degrees C over a period of 12 d. Lesion size and height of the aerial mycelium were significantly reduced by the ozone treatment on carrots inoculated with mycelial agar plugs of B. cinerea or S. sclerotiorum. Lesion size was also reduced on treated apples inoculated with 5 x 10(6) spores/mL of B. cinerea, and decay incidence of treated grapes was reduced. The 450 ppb ozone for 48 h treatment had no significant effect on color of carrots and apples or on CF of apples and grapes. Ozone, an environmentally sound antimicrobial agent, inactivates microorganisms through oxidization and residual ozone spontaneously decomposes to nontoxic products. It may be applied to fruits and vegetables to reduce decay and extend shelf life. PMID:19723209

  15. Botrytis californica, a new cryptic species in the B. cinerea species complex causing gray mold in blueberries and table grapes.

    PubMed

    Saito, S; Margosan, D; Michailides, T J; Xiao, C L

    2016-01-01

    The Botrytis cinerea species complex comprises two cryptic species, originally referred to Group I and Group II based on Bc-hch gene RFLP haplotyping. Group I was described as a new cryptic species B. pseudocinerea During a survey of Botrytis spp. causing gray mold in blueberries and table grapes in the Central Valley of California, six isolates, three from blueberries and three from table grapes, were placed in Group I but had a distinct morphological character with conidiophores significantly longer than those of B. cinerea and B. pseudocinerea We compared these with B. cinerea and B. pseudocinerea by examining morphological and physiological characters, sensitivity to fenhexamid and phylogenetic analysis inferred from sequences of three nuclear genes. Phylogenetic analysis with the three partial gene sequences encoding glyceraldehyde-3-phosate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60) and DNA-dependent RNA polymerase subunit II (RPB2) supported the proposal of a new Botrytis species, B. californica, which is closely related genetically to B. cinerea, B. pseudocinerea and B. sinoviticola, all known as causal agents of gray mold of grapes. Botrytis californica caused decay on blueberry and table grape fruit inoculated with the fungus. This study suggests that B. californica is a cryptic species sympatric with B. cinerea on blueberries and table grapes in California. PMID:26740541

  16. Effects of indole-3-acetic acid on Botrytis cinerea isolates obtained from potted plants.

    PubMed

    Martínez, J A; Valdés, R; Gómez-Bellot, M J; Bañón, S

    2011-01-01

    We study the growth of different isolates of Botrytis cinerea collected from potted plants which were affected by Botrytis blight in southern Spain during recent years. These isolates, which show widely phenotypic differences when grown in vitro, are differentially affected by growth temperature, gibberellic acid applications and paclobutrazol, an efficient plant growth retardant and fungicide at the same time. In this work, we have evaluated the effect of the auxin indole-3-acetic acid (IAA) dose (0, 1, 10, and 100 mg/plate) on the growth of the collection of B. cinerea isolates obtained from the following potted plants: Cyclamen persicum, Hydrangea macrophylla, Lantona camara, and Lonicera japonica. B. cinerea produces indolacetic acid, but so far the precise biosynthetic pathway and some effects on this fungal species are still unclear, although recent studies have revealed an antifungal activity of IAA on several fungi, including B. cinerea isolated from harvested fruits. Mycelial growth curves and growth rates assessed from difference in colony areas during the both linear and deceleration phase, conidiation (measured as time of appearance), conidia length (microm), and sclerotia production (number/plate) were evaluated in the isolates, which were grown at 26 degrees C on Petri dishes containing potato dextrose agar for up to 35 days. Mycelial growth curves fitted a typical kinetic equation of fungi grown on solid media. B. cinerea isolates showed a high degree of variability in their growth kinetics, depending on the isolate and auxin dose. This plant growth substance delayed mycelial growth during the linear phase in an isolate-dependent manner, thus isolates from C. persicum, H. macrophylla and L. camara were more affected by IAA than L. japonica. On the other hand, 100 mg of IAA was the critical dose to significantly reduce the growth rate in all isolates and to promote brown-striped hyphae development, especially in isolate from C. persicum. 10 and 100 mg

  17. Thermal inactivation of Botrytis cinerea conidia in synthetic medium and strawberry puree.

    PubMed

    Villa-Rojas, R; Sosa-Morales, M E; López-Malo, A; Tang, J

    2012-04-16

    Botrytis cinerea is one of the most important post-harvest molds that cause quality deterioration of strawberries and other fruits even during refrigeration storage. This research studied the effects of thermal inactivation of B. cinerea in synthetic medium and strawberry puree using hot water baths at different temperatures. These media were studied in order to determine if results obtained in a solution with the major components of the fruit (synthetic media), are comparable to the ones obtained in fruit purees. The results demonstrated that B. cinerea spores can be inactivated by heat treatments using relatively low temperatures (42-46 °C). Inactivation curves were well described by first order kinetics (R² 0.91-0.99). B. cinerea conidia inoculated in synthetic medium required less time to achieve one log reduction in population than those inoculated in the fruit puree. D values were 22, 8.5, 4 and 1.4 min at 42, 44, 46 and 48 °C, respectively, in synthetic medium; while D values in strawberry puree were 44.9, 13.8, 4.7 and 1.4 min at 42, 44, 46 and 48 °C, respectively. The z values obtained were 4.15 and 5.08 °C for the strawberry puree and synthetic medium respectively, showing higher sensitivity of B. cinerea in fruit purees than in the synthetic medium. Thus, a change in the medium composition had a marked difference in the heat inactivation of B. cinerea conidia, and the results obtained in synthetic medium are not accurate to describe the behavior of the microorganism in the fruit. PMID:22445202

  18. Genomic analysis of the necrotrophic fungal pathogens Sclerotinia sclerotiorum and Botrytis cinerea.

    PubMed

    Amselem, Joelle; Cuomo, Christina A; van Kan, Jan A L; Viaud, Muriel; Benito, Ernesto P; Couloux, Arnaud; Coutinho, Pedro M; de Vries, Ronald P; Dyer, Paul S; Fillinger, Sabine; Fournier, Elisabeth; Gout, Lilian; Hahn, Matthias; Kohn, Linda; Lapalu, Nicolas; Plummer, Kim M; Pradier, Jean-Marc; Quévillon, Emmanuel; Sharon, Amir; Simon, Adeline; ten Have, Arjen; Tudzynski, Bettina; Tudzynski, Paul; Wincker, Patrick; Andrew, Marion; Anthouard, Véronique; Beever, Ross E; Beffa, Rolland; Benoit, Isabelle; Bouzid, Ourdia; Brault, Baptiste; Chen, Zehua; Choquer, Mathias; Collémare, Jérome; Cotton, Pascale; Danchin, Etienne G; Da Silva, Corinne; Gautier, Angélique; Giraud, Corinne; Giraud, Tatiana; Gonzalez, Celedonio; Grossetete, Sandrine; Güldener, Ulrich; Henrissat, Bernard; Howlett, Barbara J; Kodira, Chinnappa; Kretschmer, Matthias; Lappartient, Anne; Leroch, Michaela; Levis, Caroline; Mauceli, Evan; Neuvéglise, Cécile; Oeser, Birgitt; Pearson, Matthew; Poulain, Julie; Poussereau, Nathalie; Quesneville, Hadi; Rascle, Christine; Schumacher, Julia; Ségurens, Béatrice; Sexton, Adrienne; Silva, Evelyn; Sirven, Catherine; Soanes, Darren M; Talbot, Nicholas J; Templeton, Matt; Yandava, Chandri; Yarden, Oded; Zeng, Qiandong; Rollins, Jeffrey A; Lebrun, Marc-Henri; Dickman, Marty

    2011-08-01

    Sclerotinia sclerotiorum and Botrytis cinerea are closely related necrotrophic plant pathogenic fungi notable for their wide host ranges and environmental persistence. These attributes have made these species models for understanding the complexity of necrotrophic, broad host-range pathogenicity. Despite their similarities, the two species differ in mating behaviour and the ability to produce asexual spores. We have sequenced the genomes of one strain of S. sclerotiorum and two strains of B. cinerea. The comparative analysis of these genomes relative to one another and to other sequenced fungal genomes is provided here. Their 38-39 Mb genomes include 11,860-14,270 predicted genes, which share 83% amino acid identity on average between the two species. We have mapped the S. sclerotiorum assembly to 16 chromosomes and found large-scale co-linearity with the B. cinerea genomes. Seven percent of the S. sclerotiorum genome comprises transposable elements compared to <1% of B. cinerea. The arsenal of genes associated with necrotrophic processes is similar between the species, including genes involved in plant cell wall degradation and oxalic acid production. Analysis of secondary metabolism gene clusters revealed an expansion in number and diversity of B. cinerea-specific secondary metabolites relative to S. sclerotiorum. The potential diversity in secondary metabolism might be involved in adaptation to specific ecological niches. Comparative genome analysis revealed the basis of differing sexual mating compatibility systems between S. sclerotiorum and B. cinerea. The organization of the mating-type loci differs, and their structures provide evidence for the evolution of heterothallism from homothallism. These data shed light on the evolutionary and mechanistic bases of the genetically complex traits of necrotrophic pathogenicity and sexual mating. This resource should facilitate the functional studies designed to better understand what makes these fungi such successful

  19. Homobotcinolide: a biologically active natural homolog of botcinolide from Botrytis cinerea.

    PubMed

    Cutler, H G; Parker, S R; Ross, S A; Crumley, F G; Schreiner, P R

    1996-04-01

    A novel natural product exhibiting biological activity was isolated from a strain of Botrytis cinerea that had infected raspberry fruit (Rubus ideaus). Liquid fermentation and bioassay-directed fractionation of the organism yielded a compound with molecular formula C22H38O8 that is trivially named homobotcinolide. It significantly inhibited etiolated wheat coleoptile growth. Greenhouse-grown bean, corn, and tobacco plants were also affected by exogenous application of homobotcinolide, severe chlorosis and necrosis being exhibited in corn. The compound is a polyhydroxylated nonalactone esterified with 4-hydroxy-2-decenoic acid. PMID:8829534

  20. Hexanoic acid protects tomato plants against Botrytis cinerea by priming defence responses and reducing oxidative stress.

    PubMed

    Finiti, Ivan; de la O Leyva, María; Vicedo, Begonya; Gómez-Pastor, Rocío; López-Cruz, Jaime; García-Agustín, Pilar; Real, Maria Dolores; González-Bosch, Carmen

    2014-08-01

    Treatment with the resistance priming inducer hexanoic acid (Hx) protects tomato plants from Botrytis cinerea by activating defence responses. To investigate the molecular mechanisms underlying hexanoic acid-induced resistance (Hx-IR), we compared the expression profiles of three different conditions: Botrytis-infected plants (Inf), Hx-treated plants (Hx) and Hx-treated + infected plants (Hx+Inf). The microarray analysis at 24 h post-inoculation showed that Hx and Hx+Inf plants exhibited the differential expression and priming of many Botrytis-induced genes. Interestingly, we found that the activation by Hx of other genes was not altered by the fungus at this time point. These genes may be considered to be specific targets of the Hx priming effect and may help to elucidate its mechanisms of action. It is noteworthy that, in Hx and Hx+Inf plants, there was up-regulation of proteinase inhibitor genes, DNA-binding factors, enzymes involved in plant hormone signalling and synthesis, and, remarkably, the genes involved in oxidative stress. Given the relevance of the oxidative burst occurring in plant-pathogen interactions, the effect of Hx on this process was studied in depth. We showed by specific staining that reactive oxygen species (ROS) accumulation in Hx+Inf plants was reduced and more restricted around infection sites. In addition, these plants showed higher ratios of reduced to oxidized glutathione and ascorbate, and normal levels of antioxidant activities. The results obtained indicate that Hx protects tomato plants from B. cinerea by regulating and priming Botrytis-specific and non-specific genes, preventing the harmful effects of oxidative stress produced by infection. PMID:24320938

  1. The toolbox of Trichoderma spp. in the biocontrol of Botrytis cinerea disease.

    PubMed

    Vos, Christine M F; De Cremer, Kaat; Cammue, Bruno P A; De Coninck, Barbara

    2015-05-01

    Botrytis cinerea is a necrotrophic fungal pathogen causing disease in many plant species, leading to economically important crop losses. So far, fungicides have been widely used to control this pathogen. However, in addition to their detrimental effects on the environment and potential risks for human health, increasing fungicide resistance has been observed in the B. cinerea population. Biological control, that is the application of microbial organisms to reduce disease, has gained importance as an alternative or complementary approach to fungicides. In this respect, the genus Trichoderma constitutes a promising pool of organisms with potential for B. cinerea control. In the first part of this article, we review the specific mechanisms involved in the direct interaction between the two fungi, including mycoparasitism, the production of antimicrobial compounds and enzymes (collectively called antagonism), and competition for nutrients and space. In addition, biocontrol has also been observed when Trichoderma is physically separated from the pathogen, thus implying an indirect systemic plant defence response. Therefore, in the second part, we describe the consecutive steps leading to induced systemic resistance (ISR), starting with the initial Trichoderma-plant interaction and followed by the activation of downstream signal transduction pathways and, ultimately, the defence response resulting in ISR (ISR-prime phase). Finally, we discuss the ISR-boost phase, representing the effect of ISR priming by Trichoderma spp. on plant responses after additional challenge with B. cinerea. PMID:25171761

  2. Proteome-wide analysis of lysine acetylation in the plant pathogen Botrytis cinerea.

    PubMed

    Lv, Binna; Yang, Qianqian; Li, Delong; Liang, Wenxing; Song, Limin

    2016-01-01

    Lysine acetylation is a dynamic and reversible post-translational modification that plays an important role in diverse cellular processes. Botrytis cinerea is the most thoroughly studied necrotrophic species due to its broad host range and huge economic impact. However, to date, little is known about the functions of lysine acetylation in this plant pathogen. In this study, we determined the lysine acetylome of B. cinerea through the combination of affinity enrichment and high-resolution LC-MS/MS analysis. Overall, 1582 lysine acetylation sites in 954 proteins were identified. Bioinformatics analysis shows that the acetylated proteins are involved in diverse biological functions and show multiple cellular localizations. Several particular amino acids preferred near acetylation sites, including K(ac)Y, K(ac)H, K(ac)***R, K(ac)F, FK(ac) and K(ac)***K, were identified in this organism. Protein interaction network analysis demonstrates that a variety of interactions are modulated by protein acetylation. Interestingly, 6 proteins involved in virulence of B. cinerea, including 3 key components of the high-osmolarity glycerol pathway, were found to be acetylated, suggesting that lysine acetylation plays regulatory roles in pathogenesis. These data provides the first comprehensive view of the acetylome of B. cinerea and serves as a rich resource for functional analysis of lysine acetylation in this plant pathogen. PMID:27381557

  3. Proteome-wide analysis of lysine acetylation in the plant pathogen Botrytis cinerea

    PubMed Central

    Lv, Binna; Yang, Qianqian; Li, Delong; Liang, Wenxing; Song, Limin

    2016-01-01

    Lysine acetylation is a dynamic and reversible post-translational modification that plays an important role in diverse cellular processes. Botrytis cinerea is the most thoroughly studied necrotrophic species due to its broad host range and huge economic impact. However, to date, little is known about the functions of lysine acetylation in this plant pathogen. In this study, we determined the lysine acetylome of B. cinerea through the combination of affinity enrichment and high-resolution LC-MS/MS analysis. Overall, 1582 lysine acetylation sites in 954 proteins were identified. Bioinformatics analysis shows that the acetylated proteins are involved in diverse biological functions and show multiple cellular localizations. Several particular amino acids preferred near acetylation sites, including KacY, KacH, Kac***R, KacF, FKac and Kac***K, were identified in this organism. Protein interaction network analysis demonstrates that a variety of interactions are modulated by protein acetylation. Interestingly, 6 proteins involved in virulence of B. cinerea, including 3 key components of the high-osmolarity glycerol pathway, were found to be acetylated, suggesting that lysine acetylation plays regulatory roles in pathogenesis. These data provides the first comprehensive view of the acetylome of B. cinerea and serves as a rich resource for functional analysis of lysine acetylation in this plant pathogen. PMID:27381557

  4. Postharvest Control of Botrytis cinerea and Monilinia fructigena in Apples by Gamma Irradiation Combined with Fumigation.

    PubMed

    Cheon, Wonsu; Kim, Young Soo; Balaraju, Kotnala; Kim, Bong-Su; Lee, Byeong-Ho; Jeon, Yongho

    2016-08-01

    To extend the shelf life of apples in South Korea, we evaluated the effect of gamma irradiation alone or gamma irradiation combined with fumigation on the control of postharvest decay caused by Botrytis cinerea and Monilinia fructigena. An irradiation dose of 1.0 kGy caused the maximal inhibition of B. cinerea and M. fructigena spore germination. The gamma irradiation dose required to reduce the spore germination by 90% was 0.76 and 0.78 kGy for B. cinerea and M. fructigena, respectively. Inhibition of conidial germination of both fungal pathogens occurred at a greater level at the doses of 0.2 to 1.0 kGy compared with the nontreated control; 0.2 kGy caused 90.5 and 73.9% inhibition of B. cinerea and M. fructigena, respectively. Treatment in vitro with the ecofriendly fumigant ethanedinitrile had a greater effect compared with the nontreated control. The in vitro antifungal effects of the gamma irradiation and fumigation treatments allowed us to further study the effects of the combined treatments. Interestingly, when irradiation was combined with fumigation, the percentage of disease inhibition increased more at lower (<0.4 kGy) than at higher doses of irradiation, suggesting that the combined treatments reduced the necessary irradiation dose in phytosanitary irradiation processing under storage conditions. PMID:27497129

  5. [Research of chemotaxis response of Botrytis cinerea and Alternaria panax on total ginsenosides].

    PubMed

    Chi, Kun; Xu, Yong-hua; Lei, Feng-jie; Yin, Min-jing; Wang, Zhuang; Zhang, Ai-hua; Zhang, Lian-xue

    2015-10-01

    In this paper, three kinds of chemotactic parameters (concentration, temperature and pH) were determined by plate assay and spore germination method to research the chemotactic response of Botrytis cinerea and Alternaria panax, and their spores on total ginsenosides. The results showed that Botrytis cinerea had strong chemotactic response at the mid-concentration of total ginsenosides (cultivation temperature was 20 degrees C and pH value was 6), and the data of chemotactic migration index (CMI) was 1.293 0, chemotactic growth rate (CGR) was 0.476 0, spore germination rate (SGR) was 53%, and dry weight of mycelial (DWM) was 0.452 6 g x L(-1); however, Alternaria panax had strong chemotactic response at the low-concentration of total ginsenosides (cultivation temperature was 25 degrees C and pH value was 6), and the data of chemotactic migration index (CMI) was 1.235 4, chemotactic growth rate (CGR) was 0.537 0, spore germination rate (SGR) was 67%, and dry weight of mycelial (DWM) was 0.494 8 g x L(-1). The results indicated that the low and middle concentration (2, 20 mg x L(-1)) of total ginsenosides had significant promoting effect on chemotactic response of these two pathogens, and the spore germination, mycelial growth rate, dry weight of mycelial of them were also significantly improved by this chemotactic response, whereas it decreased as the increase of total ginsenosides concentration. PMID:26975095

  6. Apoplastic Nucleoside Accumulation in Arabidopsis Leads to Reduced Photosynthetic Performance and Increased Susceptibility Against Botrytis cinerea

    PubMed Central

    Daumann, Manuel; Fischer, Marietta; Niopek-Witz, Sandra; Girke, Christopher; Möhlmann, Torsten

    2015-01-01

    Interactions between plant and pathogen often occur in the extracellular space and especially nucleotides like ATP and NAD have been identified as key players in this scenario. Arabidopsis mutants accumulating nucleosides in the extracellular space were generated and studied with respect to susceptibility against Botrytis cinerea infection and general plant fitness determined as photosynthetic performance. The mutants used are deficient in the main nucleoside uptake system ENT3 and the extracellular nucleoside hydrolase NSH3. When grown on soil but not in hydroponic culture, these plants markedly accumulate adenosine and uridine in leaves. This nucleoside accumulation was accompanied by reduced photosystem II efficiency and altered expression of photosynthesis related genes. Moreover, a higher susceptibility toward Botrytis cinerea infection and a reduced induction of pathogen related genes PR1 and WRKY33 was observed. All these effects did not occur in hydroponically grown plants substantiating a contribution of extracellular nucleosides to these effects. Whether reduced general plant fitness, altered pathogen response capability or more direct interactions with the pathogen are responsible for these observations is discussed. PMID:26779190

  7. Genomic Analysis of the Necrotrophic Fungal Pathogens Sclerotinia sclerotiorum and Botrytis cinerea

    PubMed Central

    Benito, Ernesto P.; Couloux, Arnaud; Coutinho, Pedro M.; de Vries, Ronald P.; Dyer, Paul S.; Fillinger, Sabine; Fournier, Elisabeth; Gout, Lilian; Hahn, Matthias; Kohn, Linda; Lapalu, Nicolas; Plummer, Kim M.; Pradier, Jean-Marc; Quévillon, Emmanuel; Sharon, Amir; Simon, Adeline; ten Have, Arjen; Tudzynski, Bettina; Tudzynski, Paul; Wincker, Patrick; Andrew, Marion; Anthouard, Véronique; Beffa, Rolland; Benoit, Isabelle; Bouzid, Ourdia; Brault, Baptiste; Chen, Zehua; Choquer, Mathias; Collémare, Jérome; Cotton, Pascale; Danchin, Etienne G.; Da Silva, Corinne; Gautier, Angélique; Giraud, Corinne; Giraud, Tatiana; Gonzalez, Celedonio; Grossetete, Sandrine; Güldener, Ulrich; Henrissat, Bernard; Howlett, Barbara J.; Kodira, Chinnappa; Kretschmer, Matthias; Lappartient, Anne; Leroch, Michaela; Levis, Caroline; Mauceli, Evan; Neuvéglise, Cécile; Oeser, Birgitt; Pearson, Matthew; Poulain, Julie; Poussereau, Nathalie; Quesneville, Hadi; Rascle, Christine; Schumacher, Julia; Ségurens, Béatrice; Sexton, Adrienne; Silva, Evelyn; Sirven, Catherine; Soanes, Darren M.; Talbot, Nicholas J.; Templeton, Matt; Yandava, Chandri; Yarden, Oded; Zeng, Qiandong; Rollins, Jeffrey A.; Lebrun, Marc-Henri; Dickman, Marty

    2011-01-01

    Sclerotinia sclerotiorum and Botrytis cinerea are closely related necrotrophic plant pathogenic fungi notable for their wide host ranges and environmental persistence. These attributes have made these species models for understanding the complexity of necrotrophic, broad host-range pathogenicity. Despite their similarities, the two species differ in mating behaviour and the ability to produce asexual spores. We have sequenced the genomes of one strain of S. sclerotiorum and two strains of B. cinerea. The comparative analysis of these genomes relative to one another and to other sequenced fungal genomes is provided here. Their 38–39 Mb genomes include 11,860–14,270 predicted genes, which share 83% amino acid identity on average between the two species. We have mapped the S. sclerotiorum assembly to 16 chromosomes and found large-scale co-linearity with the B. cinerea genomes. Seven percent of the S. sclerotiorum genome comprises transposable elements compared to <1% of B. cinerea. The arsenal of genes associated with necrotrophic processes is similar between the species, including genes involved in plant cell wall degradation and oxalic acid production. Analysis of secondary metabolism gene clusters revealed an expansion in number and diversity of B. cinerea–specific secondary metabolites relative to S. sclerotiorum. The potential diversity in secondary metabolism might be involved in adaptation to specific ecological niches. Comparative genome analysis revealed the basis of differing sexual mating compatibility systems between S. sclerotiorum and B. cinerea. The organization of the mating-type loci differs, and their structures provide evidence for the evolution of heterothallism from homothallism. These data shed light on the evolutionary and mechanistic bases of the genetically complex traits of necrotrophic pathogenicity and sexual mating. This resource should facilitate the functional studies designed to better understand what makes these fungi such

  8. Whole genome resequencing of Botrytis cinerea isolates identifies high levels of standing diversity.

    PubMed

    Atwell, Susanna; Corwin, Jason A; Soltis, Nicole E; Subedy, Anushryia; Denby, Katherine J; Kliebenstein, Daniel J

    2015-01-01

    How standing genetic variation within a pathogen contributes to diversity in host/pathogen interactions is poorly understood, partly because most studied pathogens are host-specific, clonally reproducing organisms which complicates genetic analysis. In contrast, Botrytis cinerea is a sexually reproducing, true haploid ascomycete that can infect a wide range of diverse plant hosts. While previous work had shown significant genomic variation between two isolates, we proceeded to assess the level and frequency of standing variation in a population of B. cinerea. To begin measuring standing genetic variation in B. cinerea, we re-sequenced the genomes of 13 different isolates and aligned them to the previously sequenced T4 reference genome. In addition one of these isolates was resequenced from four independently repeated cultures. A high level of genetic diversity was found within the 13 isolates. Within this variation, we could identify clusters of genes with major effect polymorphisms, i.e., polymorphisms that lead to a predicted functional knockout, that surrounded genes involved in controlling vegetative incompatibility. The genotype at these loci was able to partially predict the interaction of these isolates in vegetative fusion assays showing that these loci control vegetative incompatibility. This suggests that the vegetative incompatibility loci within B. cinerea are associated with regions of increased genetic diversity. The genome re-sequencing of four clones from the one isolate (Grape) that had been independently propagated over 10 years showed no detectable spontaneous mutation. This suggests that B. cinerea does not display an elevated spontaneous mutation rate. Future work will allow us to test if, and how, this diversity may be contributing to the pathogen's broad host range. PMID:26441923

  9. Whole genome resequencing of Botrytis cinerea isolates identifies high levels of standing diversity

    PubMed Central

    Atwell, Susanna; Corwin, Jason A.; Soltis, Nicole E.; Subedy, Anushryia; Denby, Katherine J.; Kliebenstein, Daniel J.

    2015-01-01

    How standing genetic variation within a pathogen contributes to diversity in host/pathogen interactions is poorly understood, partly because most studied pathogens are host-specific, clonally reproducing organisms which complicates genetic analysis. In contrast, Botrytis cinerea is a sexually reproducing, true haploid ascomycete that can infect a wide range of diverse plant hosts. While previous work had shown significant genomic variation between two isolates, we proceeded to assess the level and frequency of standing variation in a population of B. cinerea. To begin measuring standing genetic variation in B. cinerea, we re-sequenced the genomes of 13 different isolates and aligned them to the previously sequenced T4 reference genome. In addition one of these isolates was resequenced from four independently repeated cultures. A high level of genetic diversity was found within the 13 isolates. Within this variation, we could identify clusters of genes with major effect polymorphisms, i.e., polymorphisms that lead to a predicted functional knockout, that surrounded genes involved in controlling vegetative incompatibility. The genotype at these loci was able to partially predict the interaction of these isolates in vegetative fusion assays showing that these loci control vegetative incompatibility. This suggests that the vegetative incompatibility loci within B. cinerea are associated with regions of increased genetic diversity. The genome re-sequencing of four clones from the one isolate (Grape) that had been independently propagated over 10 years showed no detectable spontaneous mutation. This suggests that B. cinerea does not display an elevated spontaneous mutation rate. Future work will allow us to test if, and how, this diversity may be contributing to the pathogen's broad host range. PMID:26441923

  10. Optimisation of techniques for quantification of Botrytis cinerea in grape berries and receptacles by quantitative polymerase chain reaction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Quantitative PCR (qPCR) can be used to detect and monitor pathogen colonization, but early attempts to apply the technology to Botrytis cinerea infection of grape berries have identified limitations to current techniques. In this study, four DNA extraction methods, two grinding methods, two grape or...

  11. Botrydial and botcinins produced by Botrytis cinerea regulate expression of Trichoderma arundinaceum genes involved in trichothecene biosynthesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Trichoderma arundinaceum (Ta37) and Botrytis cinerea produce the sesquiterpenes harzianum A (HA) and botrydial (BOT), respectively, and also the polyketides aspinolides (Asp) and botcinines (Botc), respectively. In the present work, we analyzed the role of BOT and Botcs in the T. arundinaceum-B. cin...

  12. Botrydial and Botcinins produced by Botrytis cinerea regulate expression of Trichoderma arundinaceum genes involved in trichothecene biosynthesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Trichoderma arundinaceum (Ta37) and Botrytis cinerea produce the sesquiterpenes harzianum A (HA) and botrydial (BOT), respectively, and also the polyketides aspinolides (Asp) and botcinines (Botc), respectively. In the present work, we analyzed the role of BOT and Botcs in the T. arundinaceum-B. cin...

  13. [Synergistion mechanism of exogenous Ca2+ to SA-induced resistance to Botrytis cinerea in tomato].

    PubMed

    Li, Lin-lin; Li, Tian-lai; Jiang, Guo-bin; Jin, Hua; Zou, Ji-xiang

    2015-11-01

    In this study, we investigated the effect of exogenous calcium and salicylic acid (SA) on Botrytis cinerea resistance in tomato seedlings. We treated a tomato strain susceptible to Botrytis cinerea with foliar spraying of water, SA, SA+CaCl2 and SA+EGTA (Ca2+ chelating agent) for one to five days. During the treatment, leaves were collected to analyze the reactive oxygen species (ROS) content, phenylalanine ammonia lyase (PAL) activity, chintase and β-1,3-glucanase levels, and the expression of pathogenesis related protein 1, 2, 3 (PR1, PR2, PR3). Three days after infection, the disease index was 74.8 in control plants, and 46.9, 38.5 and 70.3 in SA, SA+Ca and SA+ EGTA treated plants, respectively. SA treatment significantly increased ROS leaf accumulation, and activities of PAL, chintase and β-1,3-glucanase. These values were further enhanced in SA+Ca treated plants, but decreased in SA+EGTA treated plants. Application of SA significantly increased the expression levels of PR1, PR2a and PR3b, which were further elevated by the combination treatment with Ca2+. These effects were counteracted by the combination treatment of SA and EGTA. The transcription levels of PR2b and PR3a were up-regulated by 1-2 folds, and PR1, 2a and 3b by 2-5 folds in SA- and SA+Ca-treated plants relative to control. These data suggested that application of Ca2+ could synergistically increase SA-induced resistance to B. cinerea. The resistance was associated with ROS accumulation, therefore the increase in resistance might be through ROS ability to increase the activity of defense-related enzymes and expression levels of PR1, PR2a and PR3b. PMID:26915208

  14. A circadian oscillator in the fungus Botrytis cinerea regulates virulence when infecting Arabidopsis thaliana

    PubMed Central

    Hevia, Montserrat A.; Canessa, Paulo; Müller-Esparza, Hanna; Larrondo, Luis F.

    2015-01-01

    The circadian clock of the plant model Arabidopsis thaliana modulates defense mechanisms impacting plant–pathogen interactions. Nevertheless, the effect of clock regulation on pathogenic traits has not been explored in detail. Moreover, molecular description of clocks in pathogenic fungi—or fungi in general other than the model ascomycete Neurospora crassa—has been neglected, leaving this type of question largely unaddressed. We sought to characterize, therefore, the circadian system of the plant pathogen Botrytis cinerea to assess if such oscillatory machinery can modulate its virulence potential. Herein, we show the existence of a functional clock in B. cinerea, which shares similar components and circuitry with the Neurospora circadian system, although we found that its core negative clock element FREQUENCY (BcFRQ1) serves additional roles, suggesting extracircadian functions for this protein. We observe that the lesions produced by this necrotrophic fungus on Arabidopsis leaves are smaller when the interaction between these two organisms occurs at dawn. Remarkably, this effect does not depend solely on the plant clock, but instead largely relies on the pathogen circadian system. Genetic disruption of the B. cinerea oscillator by mutation, overexpression of BcFRQ1, or by suppression of its rhythmicity by constant light, abrogates circadian regulation of fungal virulence. By conducting experiments with out-of-phase light:dark cycles, we confirm that indeed, it is the fungal clock that plays the main role in defining the outcome of the Arabidopsis–Botrytis interaction, providing to our knowledge the first evidence of a microbial clock modulating pathogenic traits at specific times of the day. PMID:26124115

  15. Combinations of Fungicides with Phylloplane Yeasts for Improved Control of Botrytis cinerea on Geranium Seedlings.

    PubMed

    Buck, J W

    2004-02-01

    ABSTRACT Control of Botrytis cinerea on geranium seedlings was evaluated in treatments with phylloplane yeasts in combination with 10 fungicides used to manage Botrytis blight of ornamental plants. Rhodotorula glutinis PM4 significantly reduced the development of lesions caused by B. cinerea on geranium cotyledons; however, yeast biocontrol efficacy was highly variable between trials. Treatment with the yeast in combination with azoxystrobin or trifloxystrobin at one tenth the labeled rate (7.5 mug a.i. ml(-1)) or the full labeled rate (7.5 mug a.i. ml(-1)) reduced lesion development, compared to treatment with the yeast or the fungicide alone. Vinclozolin at half the labeled rate or the full labeled rate (250 or 500 mug a.i. ml(-1)), in combination with R. glutinis PM4, significantly reduced the development of lesions caused by an isolate of B. cinerea resistant to vinclozolin. Copper hydroxide and iprodione at one-tenth the labeled rates, with or without yeast, were highly effective in limiting lesion development. Mancozeb did not increase the biocontrol efficacy of the yeast, and thiophanate-methyl negatively affected the yeast efficacy. Improved disease control was observed in treatments with vinclozolin at the labeled rate and R. glutinis PM4 at cell densities of 5 x 10(5) and 1 x 10(6) cells ml(-1), but not 1 x 10(5) cells ml(-1), on seedlings co-inoculated with B. cinerea in a suspension containing 1 x 10(5) conidia ml(-1). Disease control improved in treatments with combinations of vinclozolin and eight other isolates of R. glutinis, two isolates of R. graminis, and two isolates of R. mucilaginosa. Biocontrol was not observed in treatments with two isolates of R. minuta. The combination of yeast and vinclozolin significantly reduced the germination of conidia of B. cinerea and the growth of R. glutinis PM4 in vitro. All combinations of R. glutinis PM4 with azoxystrobin, trifloxystrobin, or vinclozolin provided highly effective and consistent disease control

  16. An aspartic proteinase gene family in the filamentous fungus Botrytis cinerea contains members with novel features.

    PubMed

    ten Have, Arjen; Dekkers, Ester; Kay, John; Phylip, Lowri H; van Kan, Jan A L

    2004-07-01

    Botrytis cinerea, an important fungal plant pathogen, secretes aspartic proteinase (AP) activity in axenic cultures. No cysteine, serine or metalloproteinase activity could be detected. Proteinase activity was higher in culture medium containing BSA or wheat germ extract, as compared to minimal medium. A proportion of the enzyme activity remained in the extracellular glucan sheath. AP was also the only type of proteinase activity in fluid obtained from B. cinerea-infected tissue of apple, pepper, tomato and zucchini. Five B. cinerea genes encoding an AP were cloned and denoted Bcap1-5. Features of the encoded proteins are discussed. BcAP1, especially, has novel characteristics. A phylogenetic analysis was performed comprising sequences originating from different kingdoms. BcAP1 and BcAP5 did not cluster in a bootstrap-supported clade. BcAP2 clusters with vacuolar APs. BcAP3 and BcAP4 cluster with secreted APs in a clade that also contains glycosylphosphatidylinositol-anchored proteinases from Saccharomyces cerevisiae and Candida albicans. All five Bcap genes are expressed in liquid cultures. Transcript levels of Bcap1, Bcap2, Bcap3 and Bcap4 are subject to glucose and peptone repression. Transcripts from all five Bcap genes were detected in infected plant tissue, indicating that at least part of the AP activity in planta originates from the pathogen. PMID:15256589

  17. Streptomyces araujoniae Produces a Multiantibiotic Complex with Ionophoric Properties to Control Botrytis cinerea.

    PubMed

    Silva, Leonardo José; Crevelin, Eduardo José; Souza, Wallace Rafael; Moraes, Luiz Alberto Beraldo; Melo, Itamar Soares; Zucchi, Tiago Domingues

    2014-12-01

    A recently described actinomycete species (Streptomyces araujoniae ASBV-1(T)) is effective against many phytopathogenic fungi. In this study, we evaluated the capacity of this species to inhibit Botrytis cinerea development in strawberry pseudofruit, and we identified the chemical structures of its bioactive compounds. An ethyl acetate crude extract (0.1 mg ml(-1)) of ASBV-1(T) fermentation broth completely inhibited fungus growth in strawberry pseudofruit under storage conditions. The crude extract was fractionated by preparative high-performance liquid chromatography; the active fraction was further evaluated by tandem mass spectrometry. ASBV-1(T) produced a multiantibiotic complex with ionophoric properties. This complex contained members of the macrotetralides class (including monactin, dinactin, trinactin, and tetranactin) and the cyclodepsipeptide valinomycin, all of which were active against B. cinerea. Furthermore, the addition of 2 mM MgSO4 and 1 mM ZnSO4 enhanced macrotetralide and valinomycin production, respectively, in the culture broth. These compounds are considered to be the main active molecules that S. araujoniae produces to control B. cinerea. Their low to moderate toxicity to humans and the environment justifies the application of ASBV-1(T) in biological control programs that aim to mitigate the damage caused by this phytopathogen. PMID:24983843

  18. Botrytis cinerea Protein O-Mannosyltransferases Play Critical Roles in Morphogenesis, Growth, and Virulence

    PubMed Central

    González, Mario; Brito, Nélida; Frías, Marcos; González, Celedonio

    2013-01-01

    Protein O-glycosylation is crucial in determining the structure and function of numerous secreted and membrane-bound proteins. In fungi, this process begins with the addition of a mannose residue by protein O-mannosyltransferases (PMTs) in the lumen side of the ER membrane. We have generated mutants of the three Botrytis cinerea pmt genes to study their role in the virulence of this wide-range plant pathogen. B. cinerea PMTs, especially PMT2, are critical for the stability of the cell wall and are necessary for sporulation and for the generation of the extracellular matrix. PMTs are also individually required for full virulence in a variety of hosts, with a special role in the penetration of intact plant leaves. The most significant case is that of grapevine leaves, whose penetration requires the three functional PMTs. Furthermore, PMT2 also contributes significantly to fungal adherence on grapevine and tobacco leaves. Analysis of extracellular and membrane proteins showed significant changes in the pattern of protein secretion and glycosylation by the pmt mutants, and allowed the identification of new protein substrates putatively glycosylated by specific PMTs. Since plants do no possess these enzymes, PMTs constitute a promising target in the development of novel control strategies against B. cinerea. PMID:23762450

  19. Biological characteristics and resistance analysis of the novel fungicide SYP-1620 against Botrytis cinerea.

    PubMed

    Zhang, Xiaoke; Wu, Dongxia; Duan, Yabing; Ge, Changyan; Wang, Jianxin; Zhou, Mingguo; Chen, Changjun

    2014-09-01

    SYP-1620, a quinone-outside-inhibitor (QoI), is a novel broad-spectrum fungicide. In this study, 108 isolates of Botrytis cinerea from different geographical regions in Jiangsu Province of China were characterized for baseline sensitivity to SYP-1620. The curves of baseline sensitivity were unimodal with a mean EC50 value of 0.0130±0.0109 μg/mL for mycelial growth, 0.01147±0.0062 μg/mL for spore germination, respectively. The biological characterization of SYP-1620 against B. cinerea was determined in vitro. The results indicated that SYP-1620 has a strong inhibiting effect on spore germination, mycelial growth, and respiration. The protective and curative test of SYP-1620 suggested that protective effect was better than curative either on strawberry leaves or on cucumber leaves in vivo. In addition, the biological characterization of SYP-1620-resistant mutants of B. cinerea was investigated. SYP-1620 has no cross-resistance with other types of fungicide. Compared to the sensitive isolates, the resistant mutants had lower mycelial growth and virulence but not differ in mycelial dry weight. Sequencing indicated that SYP-1620 resistance was associated with a single point mutation (G143A) in the cytochrome b gene. PMID:25175653

  20. BcIEB1, a Botrytis cinerea secreted protein, elicits a defense response in plants.

    PubMed

    Frías, Marcos; González, Mario; González, Celedonio; Brito, Nélida

    2016-09-01

    BcIEB1 is a very abundant protein in the secretome of Botrytis cinerea but it has no known function and no similarity to any characterized protein family. Previous results suggested that this protein is an elicitor of the plant defense system. In this work we have generated loss-of-function B. cinerea mutants lacking BcIEB1 and we have expressed the protein in yeast to assay its activity on plants. Analysis of the Δbcieb1 mutants did not result in any observable phenotype, including no difference in the virulence on a variety of hosts. However, when BcIEB1 was applied to plant tissues it produced necrosis as well as a whole range of symptoms: inhibition of seedling growth in Arabidopsis and tobacco, ion leakage from tobacco leaf disks, a ROS burst, cell death and autofluorescence in onion epidermis, as well as the expression of defense genes in tobacco. Moreover, tobacco plants treated with BcIEB1 showed an increased systemic resistance to B. cinerea. A small 35-amino acids peptide derived from a conserved region of BcIEB1 is almost as active on plants as the whole protein. These results clearly indicate that BcIEB1 elicits plant defenses, probably as a consequence of its recognition as a pathogen associated molecular pattern. PMID:27457989

  1. Antifungal Activity of Resveratrol against Botrytis cinerea Is Improved Using 2-Furyl Derivatives

    PubMed Central

    Caruso, Francesco; Mendoza, Leonora; Castro, Paulo; Cotoras, Milena; Aguirre, Maria; Matsuhiro, Betty; Isaacs, Mauricio; Rossi, Miriam; Viglianti, Angela; Antonioletti, Roberto

    2011-01-01

    The antifungal effect of three furyl compounds closely related to resveratrol, (E)-3,4,5-trimethoxy-β-(2-furyl)-styrene (1), (E)-4-methoxy-β-(2-furyl)-styrene (2) and (E)-3,5-dimethoxy-β-(2-furyl)-styrene (3) against Botrytis cinerea was analyzed. The inhibitory effect, at 100 µg ml−1 of compounds 1, 2, 3 and resveratrol on conidia germination, was determined to be about 70%, while at the same concentration pterostilbene (a dimethoxyl derivative of resveratrol) produced complete inhibition. The title compounds were more fungitoxic towards in vitro mycelial growth than resveratrol and pterostilbene. Compound 3 was the most active and a potential explanation of this feature is given using density functional theory (DFT) calculations on the demethoxylation/demethylation process. Compound 3 was further evaluated for its effects on laccase production, oxygen consumption and membrane integrity of B. cinerea. An increase of the laccase activity was observed in the presence of compound 3 and, using Sytox Green nucleic acid stain, it was demonstrated that this compound altered B. cinerea membrane. Finally, compound 3 partially affected conidia respiration. PMID:22022392

  2. Genome-wide transcriptional profiling of Botrytis cinerea genes targeting plant cell walls during infections of different hosts

    PubMed Central

    Blanco-Ulate, Barbara; Morales-Cruz, Abraham; Amrine, Katherine C. H.; Labavitch, John M.; Powell, Ann L. T.; Cantu, Dario

    2014-01-01

    Cell walls are barriers that impair colonization of host tissues, but also are important reservoirs of energy-rich sugars. Growing hyphae of necrotrophic fungal pathogens, such as Botrytis cinerea (Botrytis, henceforth), secrete enzymes that disassemble cell wall polysaccharides. In this work we describe the annotation of 275 putative secreted Carbohydrate-Active enZymes (CAZymes) identified in the Botrytis B05.10 genome. Using RNAseq we determined which Botrytis CAZymes were expressed during infections of lettuce leaves, ripe tomato fruit, and grape berries. On the three hosts, Botrytis expressed a common group of 229 potentially secreted CAZymes, including 28 pectin backbone-modifying enzymes, 21 hemicellulose-modifying proteins, 18 enzymes that might target pectin and hemicellulose side-branches, and 16 enzymes predicted to degrade cellulose. The diversity of the Botrytis CAZymes may be partly responsible for its wide host range. Thirty-six candidate CAZymes with secretion signals were found exclusively when Botrytis interacted with ripe tomato fruit and grape berries. Pectin polysaccharides are notably abundant in grape and tomato cell walls, but lettuce leaf walls have less pectin and are richer in hemicelluloses and cellulose. The results of this study not only suggest that Botrytis targets similar wall polysaccharide networks on fruit and leaves, but also that it may selectively attack host wall polysaccharide substrates depending on the host tissue. PMID:25232357

  3. Fungicide resistance profiling in Botrytis cinerea populations from blueberries in California and Washington and their impact on control of gray mold

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gray mold caused by Botrytis cinerea is a major postharvest disease of blueberries grown in the Central Valley of California (CA) and western Washington State (WA). Sensitivities to boscalid, cyprodinil, fenhexamid, fludioxonil, and pyraclostrobin, representing five different fungicide classes, were...

  4. Identification and Characterization of Botrytis Blossom Blight of Japanese Plums Caused by Botrytis cinerea and B. prunorum sp. nov. in Chile.

    PubMed

    Ferrada, Enrique E; Latorre, Bernardo A; Zoffoli, Juan P; Castillo, Antonio

    2016-02-01

    Blossom blight is a destructive disease of plums (Prunus salicina) when humid and temperate weather conditions occur in Chile. Disease incidence ranging from 4 to 53% has been observed. Symptoms include light brown petal necrosis, starting as light brown mottles or V-shaped necrosis at the margins of the petals, progressing to the stamen and pistils. In this study, the etiology of blossom blight of plums was determined. High- and low-sporulating isolates of Botrytis were obtained consistently from blighted blossoms and apparently healthy flowers of plums. Based on colony morphology, conidial production and molecular phylogenetic analysis, these high- and low-sporulating isolates were identified as B. cinerea and B. prunorum sp. nov., respectively. Phylogenetic analysis of the genes glyceraldehyde 3-phosphate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60), and DNA-dependent RNA polymerase subunit II (RPB2) grouped B. prunorum isolates in a single cluster, distantly from B. cinerea and other Botrytis species. The phylogenetic analysis of necrosis and ethylene-inducing protein (NEP1 and NEP2) genes corroborated these results. Analysis of the internal transcribed spacer region and large-subunit (26S) ribosomal DNA and detection of Boty and Flipper transposable elements, were not useful to differentiate between these Botrytis species. Both species were pathogenic on plum flowers and the fruit of plums, apples, and kiwifruits. However, B. prunorum was less virulent than B. cinerea. These pathogens were re-isolated from inoculated and diseased tissues; thus, Koch's postulates were fulfilled, confirming its role in blossom blight of plums. B. cinerea was predominant, suggesting that B. prunorum may play a secondary role in the epidemiology of blossom blight in plums in Chile. This study clearly demonstrated that the etiology of blossom blight of plums is caused by B. cinerea and B. prunorum, which constitute a species complex living in sympatry on plums and possibly

  5. Growth Simulation and Discrimination of Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum Using Hyperspectral Reflectance Imaging

    PubMed Central

    Gu, Xinzhe; Wang, Zhenjie; Huang, Yangmin; Wei, Yingying; Zhang, Miaomiao; Tu, Kang

    2015-01-01

    This research aimed to develop a rapid and nondestructive method to model the growth and discrimination of spoilage fungi, like Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum, based on hyperspectral imaging system (HIS). A hyperspectral imaging system was used to measure the spectral response of fungi inoculated on potato dextrose agar plates and stored at 28°C and 85% RH. The fungi were analyzed every 12 h over two days during growth, and optimal simulation models were built based on HIS parameters. The results showed that the coefficients of determination (R2) of simulation models for testing datasets were 0.7223 to 0.9914, and the sum square error (SSE) and root mean square error (RMSE) were in a range of 2.03–53.40×10−4 and 0.011–0.756, respectively. The correlation coefficients between the HIS parameters and colony forming units of fungi were high from 0.887 to 0.957. In addition, fungi species was discriminated by partial least squares discrimination analysis (PLSDA), with the classification accuracy of 97.5% for the test dataset at 36 h. The application of this method in real food has been addressed through the analysis of Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum inoculated in peaches, demonstrating that the HIS technique was effective for simulation of fungal infection in real food. This paper supplied a new technique and useful information for further study into modeling the growth of fungi and detecting fruit spoilage caused by fungi based on HIS. PMID:26642054

  6. Physcomitrella patens activates reinforcement of the cell wall, programmed cell death and accumulation of evolutionary conserved defense signals...upon Botrytis cinerea infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The moss Physcomitrella patens is an evolutionarily basal model system suitable to analyze plant defense responses activated after pathogen assault. Upon infection with the necrotroph Botrytis cinerea (B. cinerea), several defense mechanisms are induced in P. patens, including the fortification of t...

  7. Defense responses regulated by jasmonate and delayed senescence caused by ethylene receptor mutation contribute to tolerance of petunia to Botrytis cinerea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The death of cells can be a programmed event that occurs when plants are attacked by pathogens. Botrytis cinerea (B. cinerea), a model necrotrophic pathogen, triggers the host cell death response because it produces toxins. A hypersensitive reaction (HR) occurs at the site of contact. In Arabidopsis...

  8. ABA Suppresses Botrytis cinerea Elicited NO Production in Tomato to Influence H2O2 Generation and Increase Host Susceptibility

    PubMed Central

    Sivakumaran, Anushen; Akinyemi, Aderemi; Mandon, Julian; Cristescu, Simona M.; Hall, Michael A.; Harren, Frans J. M.; Mur, Luis A. J.

    2016-01-01

    Abscisic acid (ABA) production has emerged a susceptibility factor in plant-pathogen interactions. This work examined the interaction of ABA with nitric oxide (NO) in tomato following challenge with the ABA-synthesizing pathogen, Botrytis cinerea. Trace gas detection using a quantum cascade laser detected NO production within minutes of challenge with B. cinerea whilst photoacoustic laser detection detected ethylene production – an established mediator of defense against this pathogen – occurring after 6 h. Application of the NO generation inhibitor N-Nitro-L-arginine methyl ester (L-NAME) suppressed both NO and ethylene production and resistance against B. cinerea. The tomato mutant sitiens fails to accumulate ABA, shows increased resistance to B. cinerea and we noted exhibited elevated NO and ethylene production. Exogenous application of L-NAME or ABA reduced NO production in sitiens and reduced resistance to B. cinerea. Increased resistance to B. cinerea in sitiens have previously been linked to increased reactive oxygen species (ROS) generation but this was reduced in both L-NAME and ABA-treated sitiens. Taken together, our data suggests that ABA can decreases resistance to B. cinerea via reduction of NO production which also suppresses both ROS and ethylene production. PMID:27252724

  9. Abscisic Acid Determines Basal Susceptibility of Tomato to Botrytis cinerea and Suppresses Salicylic Acid-Dependent Signaling Mechanisms1

    PubMed Central

    Audenaert, Kris; De Meyer, Geert B.; Höfte, Monica M.

    2002-01-01

    Abscisic acid (ABA) is one of the plant hormones involved in the interaction between plants and pathogens. In this work, we show that tomato (Lycopersicon esculentum Mill. cv Moneymaker) mutants with reduced ABA levels (sitiens plants) are much more resistant to the necrotrophic fungus Botrytis cinerea than wild-type (WT) plants. Exogenous application of ABA restored susceptibility to B. cinerea in sitiens plants and increased susceptibility in WT plants. These results indicate that ABA plays a major role in the susceptibility of tomato to B. cinerea. ABA appeared to interact with a functional plant defense response against B. cinerea. Experiments with transgenic NahG tomato plants and benzo(1,2,3)thiadiazole-7-carbothioic acid demonstrated the importance of salicylic acid in the tomato-B. cinerea interaction. In addition, upon infection with B. cinerea, sitiens plants showed a clear increase in phenylalanine ammonia lyase activity, which was not observed in infected WT plants, indicating that the ABA levels in healthy WT tomato plants partly repress phenylalanine ammonia lyase activity. In addition, sitiens plants became more sensitive to benzo(1,2,3)thiadiazole-7-carbothioic acid root treatment. The threshold values for PR1a gene expression declined with a factor 10 to 100 in sitiens compared with WT plants. Thus, ABA appears to negatively modulate the salicylic acid-dependent defense pathway in tomato, which may be one of the mechanisms by which ABA levels determine susceptibility to B. cinerea. PMID:11842153

  10. ABA Suppresses Botrytis cinerea Elicited NO Production in Tomato to Influence H2O2 Generation and Increase Host Susceptibility.

    PubMed

    Sivakumaran, Anushen; Akinyemi, Aderemi; Mandon, Julian; Cristescu, Simona M; Hall, Michael A; Harren, Frans J M; Mur, Luis A J

    2016-01-01

    Abscisic acid (ABA) production has emerged a susceptibility factor in plant-pathogen interactions. This work examined the interaction of ABA with nitric oxide (NO) in tomato following challenge with the ABA-synthesizing pathogen, Botrytis cinerea. Trace gas detection using a quantum cascade laser detected NO production within minutes of challenge with B. cinerea whilst photoacoustic laser detection detected ethylene production - an established mediator of defense against this pathogen - occurring after 6 h. Application of the NO generation inhibitor N-Nitro-L-arginine methyl ester (L-NAME) suppressed both NO and ethylene production and resistance against B. cinerea. The tomato mutant sitiens fails to accumulate ABA, shows increased resistance to B. cinerea and we noted exhibited elevated NO and ethylene production. Exogenous application of L-NAME or ABA reduced NO production in sitiens and reduced resistance to B. cinerea. Increased resistance to B. cinerea in sitiens have previously been linked to increased reactive oxygen species (ROS) generation but this was reduced in both L-NAME and ABA-treated sitiens. Taken together, our data suggests that ABA can decreases resistance to B. cinerea via reduction of NO production which also suppresses both ROS and ethylene production. PMID:27252724

  11. Synthesis of New Hydrated Geranylphenols and in Vitro Antifungal Activity against Botrytis cinerea

    PubMed Central

    Soto, Mauricio; Espinoza, Luis; Chávez, María I.; Díaz, Katy; Olea, Andrés F.; Taborga, Lautaro

    2016-01-01

    Geranylated hydroquinones and other geranylated compounds isolated from Aplydium species have shown interesting biological activities. This fact has prompted a number of studies where geranylated phenol derivatives have been synthesized in order to assay their bioactivities. In this work, we report the synthesis of a series of new hydrated geranylphenols using two different synthetic approaches and their inhibitory effects on the mycelial growth of Botrytis cinerea. Five new hydrated geranylphenols were obtained by direct coupling reaction between geraniol and phenol in dioxane/water and using BF3·Et2O as the catalyst or by the reaction of a geranylated phenol with BF3·Et2O. Two new geranylated quinones were also obtained. The synthesis and structural elucidation of all new compounds is presented. All hydrated geranylphenols efficiently inhibit the mycelial growth of B. cinerea. Their activity is higher than that observed for non-hydrated compounds. These results indicate that structural modification on the geranyl chain brings about an enhancement of the inhibition effect of geranylated phenol derivatives. PMID:27271604

  12. Synthesis of New Hydrated Geranylphenols and in Vitro Antifungal Activity against Botrytis cinerea.

    PubMed

    Soto, Mauricio; Espinoza, Luis; Chávez, María I; Díaz, Katy; Olea, Andrés F; Taborga, Lautaro

    2016-01-01

    Geranylated hydroquinones and other geranylated compounds isolated from Aplydium species have shown interesting biological activities. This fact has prompted a number of studies where geranylated phenol derivatives have been synthesized in order to assay their bioactivities. In this work, we report the synthesis of a series of new hydrated geranylphenols using two different synthetic approaches and their inhibitory effects on the mycelial growth of Botrytis cinerea. Five new hydrated geranylphenols were obtained by direct coupling reaction between geraniol and phenol in dioxane/water and using BF₃·Et₂O as the catalyst or by the reaction of a geranylated phenol with BF₃·Et₂O. Two new geranylated quinones were also obtained. The synthesis and structural elucidation of all new compounds is presented. All hydrated geranylphenols efficiently inhibit the mycelial growth of B. cinerea. Their activity is higher than that observed for non-hydrated compounds. These results indicate that structural modification on the geranyl chain brings about an enhancement of the inhibition effect of geranylated phenol derivatives. PMID:27271604

  13. Biological control of strawberry gray mold caused by Botrytis cinerea using Bacillus licheniformis N1 formulation.

    PubMed

    Kim, Ju Hyun; Lee, Soo Hee; Kim, Choul Sung; Lim, Eun Kyung; Choi, Ki Hyuck; Kong, Hyun Gi; Kim, Dae Wook; Lee, Seon-Woo; Moon, Byung Ju

    2007-03-01

    Bacillus licheniformis N1 is a biological control agent to control gray mold diseases caused by Botrytis cinerea. Various formulations of B. licheniformis N1 were generated and evaluated for the activity to control strawberry gray mold. The wettable powder type formulation N1E was selected in pot experiments with remarkable disease control activity on both strawberry leaves and flowers. The N1E formulation contained 400 g of corn starch, 50 ml of olive oil, and 50 g of sucrose per a liter of bacterial fermentation culture. Optimum dilution of N1E to appropriately control the strawberry gray mold appeared to be 100-fold dilution in plastic house artificial infection experiments. The significant reduction of symptom development in the senescent leaves was apparent by the treatment of N1E at 100-fold dilution when N1E was applied before Bo. cinerea inoculation, but not after the inoculation. Both artificial infection experiments in a plastic house and natural infection experiments in the farm plastic house under production conditions revealed that the disease severity of gray mold on strawberry leaves and flowers was significantly reduced by N1E treatment. The disease control value of N1E on strawberry leaves was 81% under production conditions, as compared with the 61.5% conferred by a chemical fungicide, iprodione. This study suggests that our previously generated formulation of B. licheniformis N1 will be effective to control strawberry gray mold by its preventive activity. PMID:18050947

  14. Bcmimp1, a Botrytis cinerea Gene Transiently Expressed in planta, Encodes a Mitochondrial Protein

    PubMed Central

    Benito-Pescador, David; Santander, Daniela; Arranz, M.; Díaz-Mínguez, José M.; Eslava, Arturo P.; van Kan, Jan A. L.; Benito, Ernesto P.

    2016-01-01

    Botrytis cinerea is a widespread necrotrophic fungus which infects more than 200 plant species. In an attempt to characterize the physiological status of the fungus in planta and to identify genetic factors contributing to its ability to infect the host cells, a differential gene expression analysis during the interaction B. cinerea-tomato was carried out. Gene Bcmimp1 codes for a mRNA detected by differential display in the course of this analysis. During the interaction with the host, it shows a transient expression pattern with maximal expression levels during the colonization and maceration of the infected tissues. Bioinformatic analysis suggested that BCMIMP1 is an integral membrane protein located in the mitochondrial inner membrane. Co-localization experiments with a BCMIMP1-GFP fusion protein confirmed that the protein is targeted to the mitochondria. ΔBcmimp1 mutants do not show obvious phenotypic differences during saprophytic growth and their infection ability was unaltered as compared to the wild-type. Interestingly, the mutants produced increased levels of reactive oxygen species, likely as a consequence of disturbed mitochondrial function. Although Bcmimp1 expression is enhanced in planta it cannot be considered a pathogenicity factor. PMID:26952144

  15. Degradation of Extracellular β-(1,3)(1,6)-d-Glucan by Botrytis cinerea

    PubMed Central

    Stahmann, K.-Peter; Pielken, Petra; Schimz, Karl-Ludwig; Sahm, Hermann

    1992-01-01

    During growth on glucose, Botrytis cinerea produced extracellular β-(1,3)(1,6)-d-glucan (cinerean), which formed an adhering capsule and slime. After glucose was exhausted from the medium, cinereanase activity increased from <0.4 to 30 U/liter, effecting a striking loss in the viscosity of the culture. Cinerean was cleaved into glucose and gentiobiose. Gentiobiose was then hydrolyzed to glucose. While cinereanase activity was strongest in the culture supernatant, gentiobiase activity was located mainly in the cell wall fraction. The addition of extra glucose or cycloheximide prevented the cinerean degradation caused by an effect on cinereanase formation. Cinerean degradation was accompanied by microconidiation and sclerotium formation. B. cinerea was found to grow on cinerean with the latter as its single carbon and energy source. In this case, cinerean degradation occurred during hyphal growth, and no microconidiation or sclerotium formation was observed. Growth experiments with various carbon sources indicated that cinerean had a positive effect on the formation of cinerean-degrading enzymes. Images PMID:16348789

  16. The Sesquiterpene Synthase from the Botrydial Biosynthetic Gene Cluster of the Phytopathogen Botrytis cinerea

    PubMed Central

    Pinedo, Cristina; Wang, Chieh-Mei; Pradier, Jean-Marc; Dalmais, Bérengère; Choquer, Mathias; Pêcheur, Pascal Le; Morgant, Guillaume; Collado, Isidro G.; Cane, David E.; Viaud, Muriel

    2009-01-01

    The fungus Botrytis cinerea is the causal agent of the economically important gray mold disease that affects more than 200 ornamental and agriculturally important plant species. B. cinerea is a necrotrophic plant pathogen that secretes nonspecific phytotoxins, including the sesquiterpene botrydial and the polyketide botcinic acid. The region surrounding the previously characterized BcBOT1 gene has now been identified as the botrydial biosynthetic gene cluster. Five genes including BcBOT1 and BcBOT2 were shown by quantitative Reverse Transcription-PCR to be co-regulated through the calcineurin signaling pathway. Inactivation of the BcBOT2 gene, encoding a putative sesquiterpene cyclase, abolished botrydial biosynthesis, which could be restored by in trans complementation. Inactivation of BcBOT2 also resulted in over-production of botcinic acid that was observed to be strain-dependent. Recombinant BcBOT2 protein converted farnesyl diphosphate to the parent sesquiterpene of the botrydial biosynthetic pathway, the tricyclic alcohol presilphiperfolan-8β-ol. PMID:19035644

  17. Application of combined treatment for control of Botrytis cinerea in phytosanitary irradiation processing

    NASA Astrophysics Data System (ADS)

    Jung, Koo; Yoon, Minchul; Park, Hae-Jun; Youll Lee, Kwang; Jeong, Rae-Dong; Song, Beom-Seok; Lee, Ju-Woon

    2014-06-01

    Phytosanitary treatments are required to disinfest quarantine pests and pathogens in agricultural commodities. Gray mold in fruit is caused by Botrytis cinerea, which is one of the major postharvest pathogen of apple and pear. Irradiation treatment is a viable alternative for phytosanitary purposes and a useful nonchemical method for controlling pests and postharvest pathogens. An irradiation dose of over 0.4 kGy is used for the control of insects and fungal disease in fresh fruit, but a loss of firmness occurs. Combined treatments are needed to reduce the irradiation dose in phytosanitary irradiation processing. This study focuses on the application of combined treatments to reduce the loss of fruit quality when fresh fruit is irradiated for phytosanitary purposes. Comparing the antifungal activity against B. cinerea, while gamma irradiation showed no antifungal activity at a dose of 1.0 kGy, combined treatments (nano Ag particle, nano-sized silica silver) at a dose of 1.0 kGy showed the strongest antifungal activity. This study demonstrates the synergistic impacts of combined treatments in phytosanitary irradiation processing. Taken together, the combined treatments may affect reduction of fruit injury that occurred with irradiation only, meaning that the use of combined treatments with gamma irradiation is significantly effective for the preservation of fruit quality.

  18. Synthesis of N-substituted phthalimides and their antifungal activity against Alternaria solani and Botrytis cinerea.

    PubMed

    Pan, Le; Li, Xiuzhuang; Gong, Chengwen; Jin, Hui; Qin, Bo

    2016-06-01

    As organosulfur and organophosphorus agents, phaltane and phosmet are facing great challenges for the environmental contamination, mammalian toxicity and increasing resistance with long term use. It is efficient and meaningful to develop phthalimide-based alternatives with non-sulfur and non-phosphorus groups. A series of N-substituted phthalimides were synthesized and their antifungal activity against two disastrous phytopathogenic fungi, Alternaria solani and Botrytis cinerea was evaluated in vitro. Most of them showed significant antifungal activity against both of fungi, or either of them selectively. N-vinylphthalimide (4) and 8-[4-(phthalimide-2-yl) butyloxy] quinoline (13) were identified as the most promising candidates against B. cinerea and A. solani with the IC50 values of 7.92 μg/mL and 10.85 μg/mL respectively. The brief structure-activity relationships have revealed that vinyl, quinolyl, bromide alkyl and benzyl substitutions were appropriate substituents and coupling functional moieties indirectly with optimum alkyl chain was efficient to prepare phthalimides related fungicides. PMID:27079471

  19. Effect of Polymer Micelles on Antifungal Activity of Geranylorcinol Compounds against Botrytis cinerea.

    PubMed

    Taborga, Lautaro; Díaz, Katy; Olea, Andrés F; Reyes-Bravo, Paula; Flores, Mario E; Peña-Cortés, Hugo; Espinoza, Luis

    2015-08-12

    Herein, we explore the potential use of two micelle-forming block copolymers, i.e., Pluronic F-127 and poly(ethylene oxide)-b-poly(caprolactone), for application of fungicide agents. The polymer effect on the in vitro fungicide activity of a series of geranyl orcinol derivatives against Botrytis cinerea has been assessed. The results show that, for all test compounds, the incorporation into micelles, formed by Pluronic F-127, produces a great enhancement of the inhibitory effect on the growth of B. cinerea. For some compounds, at the lowest tested concentration (50 ppm), the percentage of inhibition increases significantly (from 0-10 to 80-90%) when the application is made using a polymer solution instead of an ethanol/water mixture. The synthesis and structural determination of a series of eight geranylphenols/diacetates, which were used as fungicide agents, are also discussed. These results suggest that polymer micelles are promising systems for application of crop-protecting agents. PMID:26196664

  20. Effects of Botrytis cinerea and Erysiphe necator fungi on the aroma character of grape must: A comparative approach.

    PubMed

    Lopez Pinar, Angela; Rauhut, Doris; Ruehl, Ernst; Buettner, Andrea

    2016-09-15

    Botrytis cinerea and Erysiphe necator are among the most relevant fungi in viticulture. In order to deepen our knowledge about their potential impact on wine quality, their effects on the aroma character of the initial stage of wine production, i.e. the must were studied. The main aroma compounds were determined by gas chromatography-olfactometry (GC-O) and ranked according to their relative intensities by means of aroma extract dilution analysis (AEDA). Clear differences between healthy and infected samples were observed. Botrytis cinerea had a positive impact on fruity and floral notes while several earthy smelling compounds were developed as result of the infection. Unlike in previous studies, however, we did not observe any clear differences in the quantities of earthy-mushroom-like smelling substances as result of the infection process with Erysiphe necator. PMID:27080903

  1. Control Effect and Possible Mechanism of the Natural Compound Phenazine-1-Carboxamide against Botrytis cinerea.

    PubMed

    Zhang, Ya; Wang, Chong; Su, Pin; Liao, Xiaolan

    2015-01-01

    To develop new agents against strawberry grey mould and to aid in the development of biological pesticides, we investigated the inhibitory effect of a natural compound, phenazine-1-carboxamide (PCN), against Botrytis cinerea using a growth rate assay. Additionally, indoor toxicity and the in vitro control effect of PCN were further studied to determine its potential mechanisms of action on B. cinerea. PCN was inhibitory against B. cinerea with a 50% effective concentration (EC50) of 108.12 μg/mL; the toxicity of PCN was equivalent to that of carbendazim (CBM). The best in vitro control effect of PCN against grey mould in strawberry (fruit) reached 75.32%, which was slightly higher than that of CBM. The field control effect of PCN against grey mould reached a maximum of 72.31% at a PCN concentration of 700 μg/mL, which was 1.02 times higher than that of CBM. Fungistatic activity was observed at low concentrations of PCN, while high concentrations of PCN resulted in fungicidal activity against B. cinerea. This natural compound strongly inhibited both spore and sclerotium germination of B. cinerea, with the best relative inhibition rates of 77.03% and 82.11%, respectively. The inhibitory effect of PCN on mycelial growth of B. cinerea was significant and reached levels of 87.32%. Scanning electron microscopy observations revealed that after 48 h of PCN treatment, the mycelia appeared loose, locally twisted, and folded, with exudation of contents; the mycelia was withered and twisted, with edge burrs, deformations, ruptures and a sheet-like structure. Transmission electron microscopy observations revealed that after 48 h of PCN treatment, the structure of the cell nucleus was unclear and the vacuoles had ruptured; additionally, various organelles exhibited disordered structures, there were substantial non-membrane transparent inclusions, the cells were plasmolysed, the cell walls were collapsed in some cases, and the hyphal tissue was essentially necrotic. A PCN

  2. Control Effect and Possible Mechanism of the Natural Compound Phenazine-1-Carboxamide against Botrytis cinerea

    PubMed Central

    Su, Pin; Liao, Xiaolan

    2015-01-01

    To develop new agents against strawberry grey mould and to aid in the development of biological pesticides, we investigated the inhibitory effect of a natural compound, phenazine-1-carboxamide (PCN), against Botrytis cinerea using a growth rate assay. Additionally, indoor toxicity and the in vitro control effect of PCN were further studied to determine its potential mechanisms of action on B. cinerea. PCN was inhibitory against B. cinerea with a 50% effective concentration (EC50) of 108.12 μg/mL; the toxicity of PCN was equivalent to that of carbendazim (CBM). The best in vitro control effect of PCN against grey mould in strawberry (fruit) reached 75.32%, which was slightly higher than that of CBM. The field control effect of PCN against grey mould reached a maximum of 72.31% at a PCN concentration of 700 μg/mL, which was 1.02 times higher than that of CBM. Fungistatic activity was observed at low concentrations of PCN, while high concentrations of PCN resulted in fungicidal activity against B. cinerea. This natural compound strongly inhibited both spore and sclerotium germination of B. cinerea, with the best relative inhibition rates of 77.03% and 82.11%, respectively. The inhibitory effect of PCN on mycelial growth of B. cinerea was significant and reached levels of 87.32%. Scanning electron microscopy observations revealed that after 48 h of PCN treatment, the mycelia appeared loose, locally twisted, and folded, with exudation of contents; the mycelia was withered and twisted, with edge burrs, deformations, ruptures and a sheet-like structure. Transmission electron microscopy observations revealed that after 48 h of PCN treatment, the structure of the cell nucleus was unclear and the vacuoles had ruptured; additionally, various organelles exhibited disordered structures, there were substantial non-membrane transparent inclusions, the cells were plasmolysed, the cell walls were collapsed in some cases, and the hyphal tissue was essentially necrotic. A PCN

  3. Development and evaluation of a novel and rapid detection assay for Botrytis cinerea based on loop-mediated isothermal amplification.

    PubMed

    Duan, Ya-Bing; Ge, Chang-Yan; Zhang, Xiao-Ke; Wang, Jian-Xin; Zhou, Ming-Guo

    2014-01-01

    Botrytis cinerea is a devastating plant pathogen that causes grey mould disease. In this study, we developed a visual detection method of B. cinerea based on the Bcos5 sequence using loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue dye (HNB). The LAMP reaction was optimal at 63 °C for 45 min. When HNB was added prior to amplification, samples with B. cinerea DNA developed a characteristic sky blue color after the reaction but those without DNA or with DNA of other plant pathogenic fungi did not. Results of HNB staining method were reconfirmed when LAMP products were subjected to gel electrophoresis. The detection limit of this LAMP assay for B. cinerea was 10(-3) ng µL(-1) of genomic DNA per reaction, which was 10-fold more sensitive than conventional PCR (10(-2) ng µL(-1)). Detection of the LAMP assay for inoculum of B. cinerea was possible in the inoculated tomato and strawberry petals. In the 191 diseased samples, 180 (94.2%) were confirmed as positive by LAMP, 172 (90.1%) positive by the tissue separation, while 147 (77.0%) positive by PCR. Because the LAMP assay performed well in aspects of sensitivity, specificity, repeatability, reliability, and visibility, it is suitable for rapid detection of B. cinerea in infected plant materials prior to storage and during transportation, such as cut flowers, fruits and vegetables. PMID:25329402

  4. Development and Evaluation of a Novel and Rapid Detection Assay for Botrytis cinerea Based on Loop-Mediated Isothermal Amplification

    PubMed Central

    Duan, Ya-Bing; Ge, Chang-Yan; Zhang, Xiao-Ke; Wang, Jian-Xin; Zhou, Ming-Guo

    2014-01-01

    Botrytis cinerea is a devastating plant pathogen that causes grey mould disease. In this study, we developed a visual detection method of B. cinerea based on the Bcos5 sequence using loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue dye (HNB). The LAMP reaction was optimal at 63°C for 45 min. When HNB was added prior to amplification, samples with B. cinerea DNA developed a characteristic sky blue color after the reaction but those without DNA or with DNA of other plant pathogenic fungi did not. Results of HNB staining method were reconfirmed when LAMP products were subjected to gel electrophoresis. The detection limit of this LAMP assay for B. cinerea was 10−3 ng µL−1 of genomic DNA per reaction, which was 10-fold more sensitive than conventional PCR (10−2 ng µL−1). Detection of the LAMP assay for inoculum of B. cinerea was possible in the inoculated tomato and strawberry petals. In the 191 diseased samples, 180 (94.2%) were confirmed as positive by LAMP, 172 (90.1%) positive by the tissue separation, while 147 (77.0%) positive by PCR. Because the LAMP assay performed well in aspects of sensitivity, specificity, repeatability, reliability, and visibility, it is suitable for rapid detection of B. cinerea in infected plant materials prior to storage and during transportation, such as cut flowers, fruits and vegetables. PMID:25329402

  5. Isolation and characteristics of protocatechuic acid from Paenibacillus elgii HOA73 against Botrytis cinerea on strawberry fruits.

    PubMed

    Nguyen, Xuan Hoa; Naing, Kyaw Wai; Lee, Young Seong; Moon, Jae Hak; Lee, Jeong Hyun; Kim, Kil Yong

    2015-05-01

    This study was undertaken to describe purification, identification, and characteristics of protocatechuic acid (PCA) isolated for the first time from Paenibacillus elgii HOA73 against Botrytis cinerea (the cause of gray mold disease on strawberry fruit). PCA was purified by different chromatographic techniques and identified as PCA (3,4-dihydroxybenzoic acid) by nuclear magnetic resonance and liquid chromatography-mass spectrometry analyses. PCA displayed potent antifungal activity against B. cinerea and Rhizoctonia solani. However, the antifungal activities were not sufficient to inhibit mycelial growth of Phytophthora capsici and Fusarium oxysporum. The minimum inhibitory concentration of PCA to inhibit any visible mycelial growth of both B. cinerea and R. solani was 64 µg ml(-1) . Most B. cinerea conidia displayed altered shape and absence of germination, or were degraded after treatment with 50 and 100 µg ml(-1) PCA, respectively. Moreover, gray mold formation on strawberry fruit was almost or completely inhibited by these PCA concentrations 7 days following infection with B. cinerea conidia, respectively. PCA may be a promising alternative to chemical fungicides as a potential biofungicide to prevent growth of B. cinerea in strawberry fruit disease management. PMID:25081931

  6. Gene Overexpression and RNA Silencing Tools for the Genetic Manipulation of the S-(+)-Abscisic Acid Producing Ascomycete Botrytis cinerea

    PubMed Central

    Ding, Zhong-Tao; Zhang, Zhi; Luo, Di; Zhou, Jin-Yan; Zhong, Juan; Yang, Jie; Xiao, Liang; Shu, Dan; Tan, Hong

    2015-01-01

    The phytopathogenic ascomycete Botrytis cinerea produces several secondary metabolites that have biotechnical significance and has been particularly used for S-(+)-abscisic acid production at the industrial scale. To manipulate the expression levels of specific secondary metabolite biosynthetic genes of B. cinerea with Agrobacterium tumefaciens-mediated transformation system, two expression vectors (pCBh1 and pCBg1 with different selection markers) and one RNA silencing vector, pCBSilent1, were developed with the In-Fusion assembly method. Both expression vectors were highly effective in constitutively expressing eGFP, and pCBSilent1 effectively silenced the eGFP gene in B. cinerea. Bcaba4, a gene suggested to participate in ABA biosynthesis in B. cinerea, was then targeted for gene overexpression and RNA silencing with these reverse genetic tools. The overexpression of bcaba4 dramatically induced ABA formation in the B. cinerea wild type strain Bc-6, and the gene silencing of bcaba4 significantly reduced ABA-production in an ABA-producing B. cinerea strain. PMID:25955649

  7. Gene Overexpression and RNA Silencing Tools for the Genetic Manipulation of the S-(+)-Abscisic Acid Producing Ascomycete Botrytis cinerea.

    PubMed

    Ding, Zhong-Tao; Zhang, Zhi; Luo, Di; Zhou, Jin-Yan; Zhong, Juan; Yang, Jie; Xiao, Liang; Shu, Dan; Tan, Hong

    2015-01-01

    The phytopathogenic ascomycete Botrytis cinerea produces several secondary metabolites that have biotechnical significance and has been particularly used for S-(+)-abscisic acid production at the industrial scale. To manipulate the expression levels of specific secondary metabolite biosynthetic genes of B. cinerea with Agrobacterium tumefaciens-mediated transformation system, two expression vectors (pCBh1 and pCBg1 with different selection markers) and one RNA silencing vector, pCBSilent1, were developed with the In-Fusion assembly method. Both expression vectors were highly effective in constitutively expressing eGFP, and pCBSilent1 effectively silenced the eGFP gene in B. cinerea. Bcaba4, a gene suggested to participate in ABA biosynthesis in B. cinerea, was then targeted for gene overexpression and RNA silencing with these reverse genetic tools. The overexpression of bcaba4 dramatically induced ABA formation in the B. cinerea wild type strain Bc-6, and the gene silencing of bcaba4 significantly reduced ABA-production in an ABA-producing B. cinerea strain. PMID:25955649

  8. Transcriptome and metabolome reprogramming in Vitis vinifera cv. Trincadeira berries upon infection with Botrytis cinerea.

    PubMed

    Agudelo-Romero, Patricia; Erban, Alexander; Rego, Cecília; Carbonell-Bejerano, Pablo; Nascimento, Teresa; Sousa, Lisete; Martínez-Zapater, José M; Kopka, Joachim; Fortes, Ana Margarida

    2015-04-01

    Vitis vinifera berries are sensitive towards infection by the necrotrophic pathogen Botrytis cinerea, leading to important economic losses worldwide. The combined analysis of the transcriptome and metabolome associated with fungal infection has not been performed previously in grapes or in another fleshy fruit. In an attempt to identify the molecular and metabolic mechanisms associated with the infection, peppercorn-sized fruits were infected in the field. Green and veraison berries were collected following infection for microarray analysis complemented with metabolic profiling of primary and other soluble metabolites and of volatile emissions. The results provided evidence of a reprogramming of carbohydrate and lipid metabolisms towards increased synthesis of secondary metabolites involved in plant defence, such as trans-resveratrol and gallic acid. This response was already activated in infected green berries with the putative involvement of jasmonic acid, ethylene, polyamines, and auxins, whereas salicylic acid did not seem to be involved. Genes encoding WRKY transcription factors, pathogenesis-related proteins, glutathione S-transferase, stilbene synthase, and phenylalanine ammonia-lyase were upregulated in infected berries. However, salicylic acid signalling was activated in healthy ripening berries along with the expression of proteins of the NBS-LRR superfamily and protein kinases, suggesting that the pathogen is able to shut down defences existing in healthy ripening berries. Furthermore, this study provided metabolic biomarkers of infection such as azelaic acid, a substance known to prime plant defence responses, arabitol, ribitol, 4-amino butanoic acid, 1-O-methyl- glucopyranoside, and several fatty acids that alone or in combination can be used to monitor Botrytis infection early in the vineyard. PMID:25675955

  9. Chemical Characterization of Different Sumac and Pomegranate Extracts Effective against Botrytis cinerea Rots.

    PubMed

    Romeo, Flora V; Ballistreri, Gabriele; Fabroni, Simona; Pangallo, Sonia; Nicosia, Maria Giulia Li Destri; Schena, Leonardo; Rapisarda, Paolo

    2015-01-01

    Pomegranate (Punica granatum L.) peel and sumac (Rhus coriaria L.) fruit and leaf extracts were chemically characterized and their ability to inhibit table grape (cv. Italia) rots caused by Botrytis cinerea was evaluated on artificially inoculated berries. Different extraction methods were applied and extracts were characterized through Ultra Fast High Performance Liquid Chromatography coupled to Photodiode array detector and Electrospray ionization Mass spectrometer (UPLC-PDA-ESI/MSn) for their phenol and anthocyanin contents. The concentrated pomegranate peel extract (PGE-C) was the richest in phenols (66.97 g gallic acid equivalents/kg) while the concentrated sumac extract from fruits (SUF-C) showed the highest anthocyanin amount (171.96 mg cyanidin 3-glucoside equivalents/kg). Both phenolic and anthocyanin profile of pomegranate and sumac extracts were quite different: pomegranate extract was rich in cyanidin 3-glucoside, pelargonidin 3-glucoside and ellagic acid derivatives, while sumac extract was characterized by 7-methyl-cyanidin 3-galactoside and gallic acid derivatives. The concentrated extracts from both pomegranate peel and sumac leaves significantly reduced the development of Botrytis rots. In particular, the extract from pomegranate peel completely inhibited the pathogen at different intervals of time (0, 12, and 24 h) between treatment and pathogen inoculation on fruits maintained at 22-24 °C and high relative humidity (RH). This extract may represent a valuable alternative to control postharvest fungal rots in view of its high efficacy because of the low cost of pomegranate peel, which is a waste product of processing factories. PMID:26133760

  10. Resistance to fludioxonil in Botrytis cinerea isolates from blackberry and strawberry.

    PubMed

    Li, Xingpeng; Fernández-Ortuño, Dolores; Grabke, Anja; Schnabel, Guido

    2014-07-01

    Site-specific fungicides, including the phenylpyrrole fludioxonil, are frequently used for gray mold control but are at risk for the development of resistance. In this study, field isolates that were low-resistant (LR) and moderately resistant (MR) to fludioxonil from blackberry and strawberry fields of North Carolina, South Carolina, and Virginia were characterized. Genes involved in osmoregulation, including bcsak1, BcOS4, bos5, and BRRG-1, were cloned and sequenced to detect potential target gene alterations; however, none were found. A previously described mutation (R632I) in transcription factor Mrr1, which is known to increase the expression of ATP-binding cassette transporter AtrB, was found in MR but not in sensitive (S) or LR isolates. Expression of atrB in MR isolates was ≈200-fold increased compared with an S isolate; however, 30- to 100-fold overexpression was also detected in LR isolates. Both MR isolates exhibited increased sensitivity to salt stress in the form of mycelial growth inhibition at 4% NaCl, indicating a disruption of osmoregulatory processes in those strains. However, the glycerol content was indistinguishable between S, LR, and MR isolates with and without exposure to fludioxonil, suggesting that the glycerol synthesis pathway may not be a part of the resistance mechanism in LR or MR strains. An investigation into the origin of LR and MR isolates from blackberry revealed two insertions in the mrr1 gene consistent with those found in the Botrytis clade group S. The emergence of strains overexpressing atrB in European and now in North American strawberry fields underscores the importance of this resistance mechanism for development of resistance to fludioxonil in Botrytis cinerea. PMID:24423402

  11. Transcriptome and metabolome reprogramming in Vitis vinifera cv. Trincadeira berries upon infection with Botrytis cinerea

    PubMed Central

    Agudelo-Romero, Patricia; Erban, Alexander; Rego, Cecília; Carbonell-Bejerano, Pablo; Nascimento, Teresa; Sousa, Lisete; Martínez-Zapater, José M.; Kopka, Joachim; Fortes, Ana Margarida

    2015-01-01

    Vitis vinifera berries are sensitive towards infection by the necrotrophic pathogen Botrytis cinerea, leading to important economic losses worldwide. The combined analysis of the transcriptome and metabolome associated with fungal infection has not been performed previously in grapes or in another fleshy fruit. In an attempt to identify the molecular and metabolic mechanisms associated with the infection, peppercorn-sized fruits were infected in the field. Green and veraison berries were collected following infection for microarray analysis complemented with metabolic profiling of primary and other soluble metabolites and of volatile emissions. The results provided evidence of a reprogramming of carbohydrate and lipid metabolisms towards increased synthesis of secondary metabolites involved in plant defence, such as trans-resveratrol and gallic acid. This response was already activated in infected green berries with the putative involvement of jasmonic acid, ethylene, polyamines, and auxins, whereas salicylic acid did not seem to be involved. Genes encoding WRKY transcription factors, pathogenesis-related proteins, glutathione S-transferase, stilbene synthase, and phenylalanine ammonia-lyase were upregulated in infected berries. However, salicylic acid signalling was activated in healthy ripening berries along with the expression of proteins of the NBS-LRR superfamily and protein kinases, suggesting that the pathogen is able to shut down defences existing in healthy ripening berries. Furthermore, this study provided metabolic biomarkers of infection such as azelaic acid, a substance known to prime plant defence responses, arabitol, ribitol, 4-amino butanoic acid, 1-O-methyl- glucopyranoside, and several fatty acids that alone or in combination can be used to monitor Botrytis infection early in the vineyard. PMID:25675955

  12. M233I Mutation in the β-Tubulin of Botrytis cinerea Confers Resistance to Zoxamide

    PubMed Central

    Cai, Meng; Lin, Dong; Chen, Lei; Bi, Yang; Xiao, Lu; Liu, Xi-li

    2015-01-01

    Three phenotypes were detected in 161 Botrytis cinerea field isolates, including ZoxSCarS (sensitive to zoxamide and carbendazim), ZoxSCarR (sensitive to zoxamide and resistant to carbendazim), and ZoxRCarR (resistant to zoxamide and carbendazim), but not ZoxRCarS (resistant to zoxamide and sensitive to carbendazim). The baseline sensitivity to zoxamide was determined with a mean EC50 of 0.76 μg/ml. Two stable ZoxRCarS isolates were obtained with a resistance factor of 13.28 and 20.43; there was a fitness penalty in mycelial growth rate, sporulation, virulence and sclerotium production. The results suggest that the resistance risk of B. cinerea to zoxamide is low where benzimidazoles have not been used. E198V, E198K and M233I, were detected in the β-tubulin of ZoxSCarR, ZoxRCarR, ZoxRCarS, respectively. Molecular docking indicated that position 198 in β-tubulin were targets for both zoxamide and carbendazim. The mutations at 198 prevented formation of hydrogen bonds between β-tubulin and carbendazim (E198V/K), and changed the conformation of the binding pocket of zoxamide (E198K). M233I had no effect on the binding of carbendazim but resulted in loss of a hydrogen bond between zoxamide and F200. M233 is suggested to be a unique target site for zoxamide and be very important in the function of β tubulin. PMID:26596626

  13. Synthesis, Fungicidal Activity and Mode of Action of 4-Phenyl-6-trifluoromethyl-2-aminopyrimidines against Botrytis cinerea.

    PubMed

    Liu, Chunhui; Cui, Zining; Yan, Xiaojing; Qi, Zhiqiu; Ji, Mingshan; Li, Xinghai

    2016-01-01

    Anilinopyrimidines are the main chemical agents for management of Botrytis cinerea. However, the drug resistance in fungi against this kind of compounds is very serious. To explore new potential fungicides against B. cinerea, a series of 4-phenyl-6-trifluoromethyl-2-amino-pyrimidine compounds (compounds III-1 to III-22) were synthesized, and their structures were confirmed by ¹H-NMR, IR and MS. Most of these compounds possessed excellent fungicidal activity. The compounds III-3 and III-13 showed higher fungicidal activity than the positive control pyrimethanil on fructose gelatin agar (FGA), and compound III-3 on potato dextrose agar (PDA) indicated high activity compared to the positive control cyprodinil. In vivo greenhouse results indicated that the activity of compounds III-3, III-8, and III-11 was significantly higher than that of the fungicide pyrimethanil. Scanning electron micrography (SEM) and transmission electron micrography (TEM) were applied to illustrate the mechanism of title compounds against B. cinerea. The title compounds, especially those containing a fluorine atom at the ortho-position on the benzene ring, could maintain the antifungal activity against B. cinerea, but their mechanism of action is different from that of cyprodinil. The present study lays a good foundation for us to find more efficient reagents against B. cinerea. PMID:27347910

  14. Polyamines Attenuate Ethylene-Mediated Defense Responses to Abrogate Resistance to Botrytis cinerea in Tomato1[C][W][OA

    PubMed Central

    Nambeesan, Savithri; AbuQamar, Synan; Laluk, Kristin; Mattoo, Autar K.; Mickelbart, Michael V.; Ferruzzi, Mario G.; Mengiste, Tesfaye; Handa, Avtar K.

    2012-01-01

    Transgenic tomato (Solanum lycopersicum) lines overexpressing yeast spermidine synthase (ySpdSyn), an enzyme involved in polyamine (PA) biosynthesis, were developed. These transgenic lines accumulate higher levels of spermidine (Spd) than the wild-type plants and were examined for responses to the fungal necrotrophs Botrytis cinerea and Alternaria solani, bacterial pathogen Pseudomonas syringae pv tomato DC3000, and larvae of the chewing insect tobacco hornworm (Manduca sexta). The Spd-accumulating transgenic tomato lines were more susceptible to B. cinerea than the wild-type plants; however, responses to A. solani, P. syringae, or M. sexta were similar to the wild-type plants. Exogenous application of ethylene precursors, S-adenosyl-Met and 1-aminocyclopropane-1-carboxylic acid, or PA biosynthesis inhibitors reversed the response of the transgenic plants to B. cinerea. The increased susceptibility of the ySpdSyn transgenic tomato to B. cinerea was associated with down-regulation of gene transcripts involved in ethylene biosynthesis and signaling. These data suggest that PA-mediated susceptibility to B. cinerea is linked to interference with the functions of ethylene in plant defense. PMID:22128140

  15. Impedance of the grape berry cuticle as a novel phenotypic trait to estimate resistance to Botrytis cinerea.

    PubMed

    Herzog, Katja; Wind, Rolf; Töpfer, Reinhard

    2015-01-01

    Warm and moist weather conditions during berry ripening provoke Botrytis cinerea (B. cinerea) causing notable bunch rot on susceptible grapevines with the effect of reduced yield and wine quality. Resistance donors of genetic loci to increase B. cinerea resistance are widely unknown. Promising traits of resistance are represented by physical features like the thickness and permeability of the grape berry cuticle. Sensor-based phenotyping methods or genetic markers are rare for such traits. In the present study, the simple-to-handle I-sensor was developed. The sensor enables the fast and reliable measurement of electrical impedance of the grape berry cuticles and its epicuticular waxes (CW). Statistical experiments revealed highly significant correlations between relative impedance of CW and the resistance of grapevines to B. cinerea. Thus, the relative impedance Zrel of CW was identified as the most important phenotypic factor with regard to the prediction of grapevine resistance to B. cinerea. An ordinal logistic regression analysis revealed a R2McFadden of 0.37 and confirmed the application of Zrel of CW for the prediction of bunch infection and in this way as novel phenotyping trait. Applying the I-sensor, a preliminary QTL region was identified indicating that the novel phenotypic trait is as well a valuable tool for genetic analyses. PMID:26024417

  16. Impedance of the Grape Berry Cuticle as a Novel Phenotypic Trait to Estimate Resistance to Botrytis Cinerea

    PubMed Central

    Herzog, Katja; Wind, Rolf; Töpfer, Reinhard

    2015-01-01

    Warm and moist weather conditions during berry ripening provoke Botrytis cinerea (B. cinerea) causing notable bunch rot on susceptible grapevines with the effect of reduced yield and wine quality. Resistance donors of genetic loci to increase B. cinerea resistance are widely unknown. Promising traits of resistance are represented by physical features like the thickness and permeability of the grape berry cuticle. Sensor-based phenotyping methods or genetic markers are rare for such traits. In the present study, the simple-to-handle I-sensor was developed. The sensor enables the fast and reliable measurement of electrical impedance of the grape berry cuticles and its epicuticular waxes (CW). Statistical experiments revealed highly significant correlations between relative impedance of CW and the resistance of grapevines to B. cinerea. Thus, the relative impedance Zrel of CW was identified as the most important phenotypic factor with regard to the prediction of grapevine resistance to B. cinerea. An ordinal logistic regression analysis revealed a R2McFadden of 0.37 and confirmed the application of Zrel of CW for the prediction of bunch infection and in this way as novel phenotyping trait. Applying the I-sensor, a preliminary QTL region was identified indicating that the novel phenotypic trait is as well a valuable tool for genetic analyses. PMID:26024417

  17. Effects of the origins of Botrytis cinerea on earthy aromas from grape broth media further inoculated with Penicillium expansum.

    PubMed

    Morales-Valle, H; Silva, L C; Paterson, R R M; Venâncio, A; Lima, N

    2011-08-01

    Earthy "off" aromas from wine and grape juice are highly detrimental to the production of quality grape products. These volatile compounds are produced on grapes by Botrytis cinerea, Penicillium expansum and/or a combination of P. expansum and B. cinerea strains. B. cinerea strains were isolated from different (a) vineyards in Spain and Portugal, (b) grape varieties (c) bunches (i.e., sound and botrytized) and (d) positions in the botrytized bunch (i.e., interior or exterior). A novel Headspace-Phase Microextraction (SPME) followed by Gas Chromatrography/Mass Spectrometry (GC-MS) dedicated to analyze geosmin, methylisoborneol (MIB), 1-octen-3-ol, fenchone and fenchol in grape broth medium was used. Approximately 50% of the B. cinerea strains induced detectable geosmin. One strain accumulated significant amounts of anisoles, demonstrating that this contamination might already occur in the vineyard. Strains from the interior of Cainho grape bunches induced more geosmin and hence it may be possible to reduce this volatile in wine by avoiding using these grapes in case of B. cinerea attack. PMID:21569951

  18. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules.

    PubMed

    Martínez-Hidalgo, Pilar; García, Juan M; Pozo, María J

    2015-01-01

    Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation. In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity. PMID:26388861

  19. Jasmonic acid involves in grape fruit ripening and resistant against Botrytis cinerea.

    PubMed

    Jia, Haifeng; Zhang, Cheng; Pervaiz, Tariq; Zhao, Pengcheng; Liu, Zhongjie; Wang, Baoju; Wang, Chen; Zhang, Lin; Fang, Jinggui; Qian, Jianpu

    2016-01-01

    Fruit ripening is a complex process that is regulated by a signal network. Whereas the regulatory mechanism of abscisic acid has been studied extensively in non-climacteric fruit, little is know about other signaling pathways involved in this process. In this study, we performed that plant hormone jasmonic acid plays an important role in grape fruit coloring and softening by increasing the transcription levels of several ripening-related genes, such as the color-related genes PAL1, DFR, CHI, F3H, GST, CHS, and UFGT; softening-related genes PG, PL, PE, Cell, EG1, and XTH1; and aroma-related genes Ecar, QR, and EGS. Lastly, the fruit anthocyanin, phenol, aroma, and cell wall materials were changed. Jasmonic acid positively regulated its biosynthesis pathway genes LOS, AOS, and 12-oxophytodienoate reductase (OPR) and signal pathway genes COI1 and JMT. RNA interference of grape jasmonic acid pathway gene VvAOS in strawberry fruit appeared fruit un-coloring phenotypes; exogenous jasmonic acid rescued this phenotypes. On the contrary, overexpression of grape jasmonic acid receptor VvCOI1 in the strawberry fruit accelerated the fruit-ripening process and induced some plant defense-related gene expression level. Furthermore, jasmonic acid treatment or strong jasmonic acid signal pathway in strawberry fruit make the fruit resistance against Botrytis cinerea. PMID:26498957

  20. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules

    PubMed Central

    Martínez-Hidalgo, Pilar; García, Juan M.; Pozo, María J.

    2015-01-01

    Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation. In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity. PMID:26388861

  1. Production and Characterization of Laccase from Botrytis cinerea 61-34

    PubMed Central

    Slomczynski, D.; Nakas, J. P.; Tanenbaum, S. W.

    1995-01-01

    An isolate of Botrytis cinerea (strain 61-34) constitutively expresses substantial amounts of extracellular laccase on a defined growth medium. The enzyme has been purified to homogeneity by a facile operational sequence, the last stage of which involves hydrophobic interaction chromatography. By these means, over 80 mg of laccase liter(sup-1) can be obtained from aerated fermentor reaction broths. The enzyme, with an estimated M(infr) of 74,000 and pI of 4.0, is a monomeric glycoprotein containing 49% carbohydrate predominantly as hexose. With 2,6-dimethoxyphenol, it exhibits a pH optimum of 3.5 and a temperature optimum of 60(deg)C, and its K(infm) is 100 (mu)M. The purified enzyme with this substrate has a specific activity of 9.1 mkat mg of protein(sup-1). Taken together with a broad substrate range and its stability in 4% sodium dodecyl sulfate or 2 M urea solutions, several biotechnology transfers are suggested. PMID:16534974

  2. Rapid isolation of mycoviral double-stranded RNA from Botrytis cinerea and Saccharomyces cerevisiae

    PubMed Central

    2011-01-01

    Background In most of the infected fungi, the mycoviruses are latent or cryptic, the infected fungus does not show disease symptoms, and it is phenotypically identical to a non-infected strain of the same species. Because of these properties, the initial stage in the search for fungi infected with mycoviruses is the detection of their viral genome, which in most of the described cases corresponds to double-stranded RNA (dsRNA). So to analyze a large number of fungal isolates it is necessary to have a simple and rapid method to detect dsRNA. Results A rapid method to isolate dsRNA from a virus-infected filamentous fungus, Botrytis cinerea, and from a killer strain of Saccharomyces cerevisiae using commercial minicolumns packed with CF11 cellulose was developed. In addition to being a rapid method, it allows to use small quantities of yeasts or mycelium as starting material, being obtained sufficient dsRNA quantity that can later be analyzed by agarose gel electrophoresis, treated with enzymes for its partial characterization, amplified by RT-PCR and cloned in appropriate vectors for further sequencing. Conclusions The method yields high quality dsRNA, free from DNA and ssRNA. The use of nucleases to degrade the DNA or the ssRNA is not required, and it can be used to isolate dsRNA from any type of fungi or any biological sample that contains dsRNA. PMID:21262001

  3. Novel hypovirulence-associated RNA mycovirus in the plant-pathogenic fungus Botrytis cinerea: molecular and biological characterization.

    PubMed

    Yu, Lin; Sang, Wen; Wu, Ming-De; Zhang, Jing; Yang, Long; Zhou, Ying-Jun; Chen, Wei-Dong; Li, Guo-Qing

    2015-04-01

    Botrytis cinerea is a pathogenic fungus causing gray mold on numerous economically important crops and ornamental plants. This study was conducted to characterize the biological and molecular features of a novel RNA mycovirus, Botrytis cinerea RNA virus 1 (BcRV1), in the hypovirulent strain BerBc-1 of B. cinerea. The genome of BcRV1 is 8,952 bp long with two putative overlapped open reading frames (ORFs), ORF1 and ORF2, coding for a hypothetical polypeptide (P1) and RNA-dependent RNA polymerase (RdRp), respectively. A -1 frameshifting region (designated the KNOT element) containing a shifty heptamer, a heptanucleotide spacer, and an H-type pseudoknot was predicted in the junction region of ORF1 and ORF2. The -1 frameshifting role of the KNOT element was experimentally confirmed through determination of the production of the fusion protein red fluorescent protein (RFP)-green fluorescent protein (GFP) by the plasmid containing the construct dsRed-KNOT-eGFP in Escherichia coli. BcRV1 belongs to a taxonomically unassigned double-stranded RNA (dsRNA) mycovirus group. It is closely related to grapevine-associated totivirus 2 and Sclerotinia sclerotiorum nonsegmented virus L. BcRV1 in strain BerBc-1 was found capable of being transmitted vertically through macroconidia and horizontally to other B. cinerea strains through hyphal contact. The presence of BcRV1 was found to be positively correlated with hypovirulence in B. cinerea, with the attenuation effects of BcRV1 on mycelial growth and pathogenicity being greatly affected by the accumulation level of BcRV1. PMID:25595766

  4. Novel Hypovirulence-Associated RNA Mycovirus in the Plant-Pathogenic Fungus Botrytis cinerea: Molecular and Biological Characterization

    PubMed Central

    Yu, Lin; Sang, Wen; Wu, Ming-De; Zhang, Jing; Yang, Long; Zhou, Ying-Jun; Chen, Wei-Dong

    2015-01-01

    Botrytis cinerea is a pathogenic fungus causing gray mold on numerous economically important crops and ornamental plants. This study was conducted to characterize the biological and molecular features of a novel RNA mycovirus, Botrytis cinerea RNA virus 1 (BcRV1), in the hypovirulent strain BerBc-1 of B. cinerea. The genome of BcRV1 is 8,952 bp long with two putative overlapped open reading frames (ORFs), ORF1 and ORF2, coding for a hypothetical polypeptide (P1) and RNA-dependent RNA polymerase (RdRp), respectively. A −1 frameshifting region (designated the KNOT element) containing a shifty heptamer, a heptanucleotide spacer, and an H-type pseudoknot was predicted in the junction region of ORF1 and ORF2. The −1 frameshifting role of the KNOT element was experimentally confirmed through determination of the production of the fusion protein red fluorescent protein (RFP)-green fluorescent protein (GFP) by the plasmid containing the construct dsRed-KNOT-eGFP in Escherichia coli. BcRV1 belongs to a taxonomically unassigned double-stranded RNA (dsRNA) mycovirus group. It is closely related to grapevine-associated totivirus 2 and Sclerotinia sclerotiorum nonsegmented virus L. BcRV1 in strain BerBc-1 was found capable of being transmitted vertically through macroconidia and horizontally to other B. cinerea strains through hyphal contact. The presence of BcRV1 was found to be positively correlated with hypovirulence in B. cinerea, with the attenuation effects of BcRV1 on mycelial growth and pathogenicity being greatly affected by the accumulation level of BcRV1. PMID:25595766

  5. Botrydial and botcinins produced by Botrytis cinerea regulate the expression of Trichoderma arundinaceum genes involved in trichothecene biosynthesis.

    PubMed

    Malmierca, Mónica G; Izquierdo-Bueno, Inmaculada; Mccormick, Susan P; Cardoza, Rosa E; Alexander, Nancy J; Moraga, Javier; Gomes, Eriston V; Proctor, Robert H; Collado, Isidro G; Monte, Enrique; Gutiérrez, Santiago

    2016-09-01

    Trichoderma arundinaceum IBT 40837 (Ta37) and Botrytis cinerea produce the sesquiterpenes harzianum A (HA) and botrydial (BOT), respectively, and also the polyketides aspinolides and botcinins (Botcs), respectively. We analysed the role of BOT and Botcs in the Ta37-B. cinerea interaction, including the transcriptomic changes in the genes involved in HA (tri) and ergosterol biosynthesis, as well as changes in the level of HA and squalene-ergosterol. We found that, when confronted with B. cinerea, the tri biosynthetic genes were up-regulated in all dual cultures analysed, but at higher levels when Ta37 was confronted with the BOT non-producer mutant bcbot2Δ. The production of HA was also higher in the interaction area with this mutant. In Ta37-bcbot2Δ confrontation experiments, the expression of the hmgR gene, encoding the 3-hydroxy-3-methylglutaryl coenzyme A reductase, which is the first enzyme of the terpene biosynthetic pathway, was also up-regulated, resulting in an increase in squalene production compared with the confrontation with B. cinerea B05.10. Botcs had an up-regulatory effect on the tri biosynthetic genes, with BotcA having a stronger effect than BotcB. The results indicate that the interaction between Ta37 and B. cinerea exerts a stimulatory effect on the expression of the tri biosynthetic genes, which, in the interaction zone, can be attenuated by BOT produced by B. cinerea B05.10. The present work provides evidence for a metabolic dialogue between T. arundinaceum and B. cinerea that is mediated by sesquiterpenes and polyketides, and that affects the outcome of the interaction of these fungi with each other and their environment. PMID:26575202

  6. Tomato SlMKK2 and SlMKK4 contribute to disease resistance against Botrytis cinerea

    PubMed Central

    2014-01-01

    Background Mitogen-activated protein kinase (MAPK) cascades are highly conserved signaling modules that mediate the transduction of extracellular stimuli via receptors/sensors into intracellular responses and play key roles in plant immunity against pathogen attack. However, the function of tomato MAPK kinases, SlMKKs, in resistance against Botrytis cinerea remains unclear yet. Results A total of five SlMKK genes with one new member, SlMKK5, were identified in tomato. qRT-PCR analyses revealed that expression of SlMKK2 and SlMKK4 was strongly induced by B. cinerea and by jasmonic acid and ethylene precursor 1-amino cyclopropane-1-carboxylic acid. Virus-induced gene silencing (VIGS)-based knockdown of individual SlMKKs and disease assays identified that SlMKK2 and SlMKK4 but not other three SlMKKs (SlMKK1, SlMKK3 and SlMKK5) are involved in resistance against B. cinerea. Silencing of SlMKK2 or SlMKK4 resulted in reduced resistance to B. cinerea, increased accumulation of reactive oxygen species and attenuated expression of defense genes after infection of B. cinerea in tomato plants. Furthermore, transient expression of constitutively active phosphomimicking forms SlMKK2DD and SlMKK4DD in leaves of Nicotiana benthamiana plants led to enhanced resistance to B. cinerea and elevated expression of defense genes. Conclusions VIGS-based knockdown of SlMKK2 and SlMKK4 expression in tomato and gain-of-function transient expression of constitutively active phosphomimicking forms SlMKK2DD and SlMKK2DD in N. benthamiana demonstrate that both SlMKK2 and SlMKK4 function as positive regulators of defense response against B. cinerea. PMID:24930014

  7. Novel Fungitoxicity Assays for Inhibition of Germination-Associated Adhesion of Botrytis cinerea and Puccinia recondita Spores

    PubMed Central

    Slawecki, Richard A.; Ryan, Eileen P.; Young, David H.

    2002-01-01

    Botrytis cinerea and Puccinia recondita spores adhere strongly to polystyrene microtiter plates coincident with germination. We developed assays for inhibition of spore adhesion in 96-well microtiter plates by using sulforhodamine B staining to quantify the adherent spores. In both organisms, fungicides that inhibited germination strongly inhibited spore adhesion, with 50% effective concentrations (EC50s) comparable to those for inhibition of germination. In contrast, fungicides that acted after germination in B. cinerea inhibited spore adhesion to microtiter plates only at concentrations much higher than their EC50s for inhibition of mycelial growth. Similarly, in P. recondita the ergosterol biosynthesis inhibitors myclobutanil and fenbuconazole acted after germination and did not inhibit spore adhesion. The assays provide a rapid, high-throughput alternative to traditional spore germination assays and may be applicable to other fungi. PMID:11823196

  8. Vitis vinifera canes, a new source of antifungal compounds against Plasmopara viticola, Erysiphe necator, and Botrytis cinerea.

    PubMed

    Schnee, Sylvain; Queiroz, Emerson F; Voinesco, Francine; Marcourt, Laurence; Dubuis, Pierre-Henri; Wolfender, Jean-Luc; Gindro, Katia

    2013-06-12

    Methanolic and ethanolic crude extracts of Vitis vinifera canes exhibited significant antifungal activity against the three major fungal pathogens affecting grapevines, Plasmopara viticola, Erysiphe necator and Botrytis cinerea. The active extracts were analyzed by LC-PDA-ESI-MS, and selected compounds were identified. Efficient targeted isolation using medium-pressure liquid chromatography afforded six pure constituents in one step. The structures of the isolated compounds were elucidated by NMR and HRMS. Six identified compounds (ampelopsin A, hopeaphenol, trans-resveratrol, ampelopsin H, ε-viniferin, and E-vitisin B) presented antifungal activities against P. viticola. ε-Viniferin also exhibited a low antifungal activity against B. cinerea. None of the identified compounds inhibited the germination of E. necator. The potential to develop a novel natural fungicide against the three major fungal pathogens affecting V. vinifera from viticulture waste material is discussed. PMID:23730921

  9. Sympatric genetic differentiation of a generalist pathogenic fungus, Botrytis cinerea, on two different host plants, grapevine and bramble.

    PubMed

    Fournier, E; Giraud, T

    2008-01-01

    Prime candidates for sympatric ecological divergence include parasites that differentiate via host shifts, because different host species exert strong disruptive selection and because both hosts and parasites are continually co-evolving. Sympatric divergence may be fostered even more strongly in phytopathogenic fungi, in particular those where sex must occur on the host, which allows adaptation alone to restrict gene flow between populations developing on different hosts. We sampled populations of Botrytis cinerea, a generalist ascomycete fungus, on sympatric grapes and brambles in six regions in France. Microsatellite data were analyzed using standard population genetics, a population graph analysis and a Bayesian approach. In addition to confirming that B. cinerea reproduces sexually, our results showed that the fungal populations on the two hosts were significantly differentiated, indicating restricted gene flow, even in sympatry. In contrast, only weak geographical differentiation could be detected. These results support the possibility of sympatric divergence associated with host use in generalist parasites. PMID:18028352

  10. An Interspecies Comparative Analysis of the Predicted Secretomes of the Necrotrophic Plant Pathogens Sclerotinia sclerotiorum and Botrytis cinerea

    PubMed Central

    2015-01-01

    Phytopathogenic fungi form intimate associations with host plant species and cause disease. To be successful, fungal pathogens communicate with a susceptible host through the secretion of proteinaceous effectors, hydrolytic enzymes and metabolites. Sclerotinia sclerotiorum and Botrytis cinerea are economically important necrotrophic fungal pathogens that cause disease on numerous crop species. Here, a powerful bioinformatics pipeline was used to predict the refined S. sclerotiorum and B. cinerea secretomes, identifying 432 and 499 proteins respectively. Analyses focusing on S. sclerotiorum revealed that 16% of the secretome encoding genes resided in small, sequence heterogeneous, gene clusters that were distributed over 13 of the 16 predicted chromosomes. Functional analyses highlighted the importance of plant cell hydrolysis, oxidation-reduction processes and the redox state to the S. sclerotiorum and B. cinerea secretomes and potentially host infection. Only 8% of the predicted proteins were distinct between the two secretomes. In contrast to S. sclerotiorum, the B. cinerea secretome lacked CFEM- or LysM-containing proteins. The 115 fungal and oomycete genome comparison identified 30 proteins specific to S. sclerotiorum and B. cinerea, plus 11 proteins specific to S. sclerotiorum and 32 proteins specific to B. cinerea. Expressed sequence tag (EST) and proteomic analyses showed that 246 S. sclerotiorum secretome encoding genes had EST support, including 101 which were only expressed in vitro and 49 which were only expressed in planta, whilst 42 predicted proteins were experimentally proven to be secreted. These detailed in silico analyses of two important necrotrophic pathogens will permit informed choices to be made when candidate effector proteins are selected for function analyses in planta. PMID:26107498

  11. Identification of miRNAs Responsive to Botrytis cinerea in Herbaceous Peony (Paeonia lactiflora Pall.) by High-Throughput Sequencing

    PubMed Central

    Zhao, Daqiu; Gong, Saijie; Hao, Zhaojun; Tao, Jun

    2015-01-01

    Herbaceous peony (Paeonia lactiflora Pall.), one of the world’s most important ornamental plants, is highly susceptible to Botrytis cinerea, and improving resistance to this pathogenic fungus is a problem yet to be solved. MicroRNAs (miRNAs) play an essential role in resistance to B. cinerea, but until now, no studies have been reported concerning miRNAs induction in P. lactiflora. Here, we constructed and sequenced two small RNA (sRNA) libraries from two B. cinerea-infected P. lactiflora cultivars (“Zifengyu” and “Dafugui”) with significantly different levels of resistance to B. cinerea, using the Illumina HiSeq 2000 platform. From the raw reads generated, 4,592,881 and 5,809,796 sRNAs were obtained, and 280 and 306 miRNAs were identified from “Zifengyu” and “Dafugui”, respectively. A total of 237 conserved and 7 novel sequences of miRNAs were differentially expressed between the two cultivars, and we predicted and annotated their potential target genes. Subsequently, 7 differentially expressed candidate miRNAs were screened according to their target genes annotated in KEGG pathways, and the expression patterns of miRNAs and corresponding target genes were elucidated. We found that miR5254, miR165a-3p, miR3897-3p and miR6450a might be involved in the P. lactiflora response to B. cinerea infection. These results provide insight into the molecular mechanisms responsible for resistance to B. cinerea in P. lactiflora. PMID:26393656

  12. Independent Emergence of Resistance to Seven Chemical Classes of Fungicides in Botrytis cinerea.

    PubMed

    Fernández-Ortuño, Dolores; Grabke, Anja; Li, Xingpeng; Schnabel, Guido

    2015-04-01

    Gray mold, caused by the fungal pathogen Botrytis cinerea, is one of the most destructive diseases of small fruit crops and control is largely dependent on the application of fungicides. As part of a region-wide resistance-monitoring program that investigated 1,890 B. cinerea isolates from 189 fields in 10 states of the United States, we identified seven isolates (0.4%) from five locations in four different states with unprecedented resistance to all seven Fungicide Resistance Action Committee (FRAC) codes with single-site modes of action including FRAC 1, 2, 7, 9, 11, 12, and 17 registered in the United States for gray mold control. Resistance to thiophanate-methyl, iprodione, boscalid, pyraclostrobin, and fenhexamid was based on target gene mutations that conferred E198A and F200Y in β-tubulin, I365N/S in Bos1, H272R/Y in SdhB, G143A in Cytb, and T63I and F412S in Erg27. Isolates were grouped into MDR1 and MDR1h phenotypes based on sensitivity to fludioxonil and variations in transcription factor mrr1. MDR1h isolates had a previously described 3-bp deletion at position 497 in mrr1. Expression of ABC transporter atrB was increased in MDR1 isolates but highest in MDR1h isolates. None of the isolates with seven single resistances (SR) had identical nucleotide variations in target genes, indicating that they emerged independently. Multifungicide resistance phenotypes did not exhibit significant fitness penalties for the parameters used in this study, but MDR1h isolates produced more sclerotia at low temperatures and exhibited increased sensitivity to salt stress. In this study we show that current resistance management strategies have not been able to prevent the geographically independent development of resistance to all seven site-specific fungicides currently registered for gray mold control in the United States and document the presence of MDR1h in North America. PMID:25317841

  13. Botrytis cinerea Manipulates the Antagonistic Effects between Immune Pathways to Promote Disease Development in Tomato[C][W][OA

    PubMed Central

    El Oirdi, Mohamed; El Rahman, Taha Abd; Rigano, Luciano; El Hadrami, Abdelbasset; Rodriguez, María Cecilia; Daayf, Fouad; Vojnov, Adrian; Bouarab, Kamal

    2011-01-01

    Plants have evolved sophisticated mechanisms to sense and respond to pathogen attacks. Resistance against necrotrophic pathogens generally requires the activation of the jasmonic acid (JA) signaling pathway, whereas the salicylic acid (SA) signaling pathway is mainly activated against biotrophic pathogens. SA can antagonize JA signaling and vice versa. Here, we report that the necrotrophic pathogen Botrytis cinerea exploits this antagonism as a strategy to cause disease development. We show that B. cinerea produces an exopolysaccharide, which acts as an elicitor of the SA pathway. In turn, the SA pathway antagonizes the JA signaling pathway, thereby allowing the fungus to develop its disease in tomato (Solanum lycopersicum). SA-promoted disease development occurs through Nonexpressed Pathogen Related1. We also show that the JA signaling pathway required for tomato resistance against B. cinerea is mediated by the systemin elicitor. These data highlight a new strategy used by B. cinerea to overcome the plant’s defense system and to spread within the host. PMID:21665999

  14. Emerging Trends in Molecular Interactions between Plants and the Broad Host Range Fungal Pathogens Botrytis cinerea and Sclerotinia sclerotiorum.

    PubMed

    Mbengue, Malick; Navaud, Olivier; Peyraud, Rémi; Barascud, Marielle; Badet, Thomas; Vincent, Rémy; Barbacci, Adelin; Raffaele, Sylvain

    2016-01-01

    Fungal plant pathogens are major threats to food security worldwide. Sclerotinia sclerotiorum and Botrytis cinerea are closely related Ascomycete plant pathogens causing mold diseases on hundreds of plant species. There is no genetic source of complete plant resistance to these broad host range pathogens known to date. Instead, natural plant populations show a continuum of resistance levels controlled by multiple genes, a phenotype designated as quantitative disease resistance. Little is known about the molecular mechanisms controlling the interaction between plants and S. sclerotiorum and B. cinerea but significant advances were made on this topic in the last years. This minireview highlights a selection of nine themes that emerged in recent research reports on the molecular bases of plant-S. sclerotiorum and plant-B. cinerea interactions. On the fungal side, this includes progress on understanding the role of oxalic acid, on the study of fungal small secreted proteins. Next, we discuss the exchanges of small RNA between organisms and the control of cell death in plant and fungi during pathogenic interactions. Finally on the plant side, we highlight defense priming by mechanical signals, the characterization of plant Receptor-like proteins and the hormone abscisic acid in the response to B. cinerea and S. sclerotiorum, the role of plant general transcription machinery and plant small bioactive peptides. These represent nine trends we selected as remarkable in our understanding of fungal molecules causing disease and plant mechanisms associated with disease resistance to two devastating broad host range fungi. PMID:27066056

  15. Unraveling the in vitro secretome of the phytopathogen Botrytis cinerea to understand the interaction with its hosts

    PubMed Central

    González-Fernández, Raquel; Valero-Galván, José; Gómez-Gálvez, Francisco J.; Jorrín-Novo, Jesús V.

    2015-01-01

    Botrytis cinerea is a necrotrophic fungus with high adaptability to different environments and hosts. It secretes a large number of extracellular proteins, which favor plant tissue penetration and colonization, thus contributing to virulence. Secretomics is a proteomics sub-discipline which study the secreted proteins and their secretion mechanisms, so-called secretome. By using proteomics as experimental approach, many secreted proteins by B. cinerea have been identified from in vitro experiments, and belonging to different functional categories: (i) cell wall-degrading enzymes such as pectinesterases and endo-polygalacturonases; (ii) proteases involved in host protein degradation such as an aspartic protease; (iii) proteins related to the oxidative burst such as glyoxal oxidase; (iv) proteins which may induce the plant hypersensitive response such as a cerato-platanin domain-containing protein; and (v) proteins related to production and secretion of toxins such as malate dehydrogenase. In this mini-review, we made an overview of the proteomics contribution to the study and knowledge of the B. cinerea extracellular secreted proteins based on our current work carried out from in vitro experiments, and recent published papers both in vitro and in planta studies on this fungi. We hypothesize on the putative functions of these secreted proteins, and their connection to the biology of the B. cinerea interaction with its hosts. PMID:26500673

  16. Emerging Trends in Molecular Interactions between Plants and the Broad Host Range Fungal Pathogens Botrytis cinerea and Sclerotinia sclerotiorum

    PubMed Central

    Mbengue, Malick; Navaud, Olivier; Peyraud, Rémi; Barascud, Marielle; Badet, Thomas; Vincent, Rémy; Barbacci, Adelin; Raffaele, Sylvain

    2016-01-01

    Fungal plant pathogens are major threats to food security worldwide. Sclerotinia sclerotiorum and Botrytis cinerea are closely related Ascomycete plant pathogens causing mold diseases on hundreds of plant species. There is no genetic source of complete plant resistance to these broad host range pathogens known to date. Instead, natural plant populations show a continuum of resistance levels controlled by multiple genes, a phenotype designated as quantitative disease resistance. Little is known about the molecular mechanisms controlling the interaction between plants and S. sclerotiorum and B. cinerea but significant advances were made on this topic in the last years. This minireview highlights a selection of nine themes that emerged in recent research reports on the molecular bases of plant-S. sclerotiorum and plant-B. cinerea interactions. On the fungal side, this includes progress on understanding the role of oxalic acid, on the study of fungal small secreted proteins. Next, we discuss the exchanges of small RNA between organisms and the control of cell death in plant and fungi during pathogenic interactions. Finally on the plant side, we highlight defense priming by mechanical signals, the characterization of plant Receptor-like proteins and the hormone abscisic acid in the response to B. cinerea and S. sclerotiorum, the role of plant general transcription machinery and plant small bioactive peptides. These represent nine trends we selected as remarkable in our understanding of fungal molecules causing disease and plant mechanisms associated with disease resistance to two devastating broad host range fungi. PMID:27066056

  17. Fitness and Competitive Ability of Botrytis cinerea Isolates with Resistance to Multiple Chemical Classes of Fungicides.

    PubMed

    Chen, S N; Luo, C X; Hu, M J; Schnabel, G

    2016-09-01

    Resistance to multiple chemical classes of fungicides in Botrytis cinerea isolates from eastern United States strawberry fields is common and strategies to control them are needed. In this study, we compared fitness and competitive ability of eight sensitive isolates (S), eight isolates resistant to five or six chemical classes of fungicides but not to phenylpyrroles (5CCR), and eight isolates resistant to six or seven chemical classes including phenylpyrroles (6CCR/MDR1h). The latter included the MDR1h phenotype due to overexpression of atrB based on Δ497V/L in mrr1. The 6CCR/MDR1h isolates grew more slowly at 4°C on potato dextrose agar, and both 5CCR and 6CCR/MDR1h isolates were hypersensitive to osmotic stress compared with S isolates. In contrast, no differences were found in oxidative sensitivity, aggressiveness, and spore production in vivo, and sclerotia production and viability in vitro. In competition experiments, the 5CCR and 6CCR/MDR1h isolates were both outcompeted by S isolates and 6CCR/MDR1h isolates were outcompeted by 5CCR isolates in the absence of fungicide pressure. Under selective pressure of a fludioxonil/pyraclostrobin rotation, the 6CCR/MDR1h isolates dominated after coinoculation with 5CCR and S isolates. The competitive disadvantage of 5CCR and especially 6CCR/MDR1h isolates suggest that, in the absence of fungicide selection pressure, S isolates may reduce inoculum potential of multifungicide-resistant isolates under field conditions. PMID:27161219

  18. Microfluidic immunosensor with micromagnetic beads coupled to carbon-based screen-printed electrodes (SPCEs) for determination of Botrytis cinerea in tissue of fruits.

    PubMed

    Fernández-Baldo, Martín A; Messina, Germán A; Sanz, Maria I; Raba, Julio

    2010-11-10

    A wide range of plant species, including economically important crops such as vegetables, ornamentals, bulbs, and fundamentally fruits, can be affected by gray mold caused by the fungal pathogen Botrytis cinerea . This paper describes the development of a microfluidic immunosensor with micromagnetic beads (MMBs) coupled to carbon-based screen-printed electrodes (SPCEs) for the rapid and sensitive quantification of B. cinerea in apple (Red Delicious), table grape (pink Moscatel), and pear (William's) tissues. The detection of B. cinerea was carried out using a competitive immunoassay method based on the use of purified B. cinerea antigens immobilized on 3-aminopropyl-modified MMBs. The total assay time was 40 min, and the calculated detection limit was 0.008 μg mL(-1). Moreover, the intra- and interassay coefficients of variation were below 7%. The developed method allowed detects B. cinerea even in asymptomatic fruits and promises to be particularly useful for application in the agricultural industry. PMID:20931959

  19. A Mechanistic Model of Botrytis cinerea on Grapevines That Includes Weather, Vine Growth Stage, and the Main Infection Pathways.

    PubMed

    González-Domínguez, Elisa; Caffi, Tito; Ciliberti, Nicola; Rossi, Vittorio

    2015-01-01

    A mechanistic model for Botrytis cinerea on grapevine was developed. The model, which accounts for conidia production on various inoculum sources and for multiple infection pathways, considers two infection periods. During the first period ("inflorescences clearly visible" to "berries groat-sized"), the model calculates: i) infection severity on inflorescences and young clusters caused by conidia (SEV1). During the second period ("majority of berries touching" to "berries ripe for harvest"), the model calculates: ii) infection severity of ripening berries by conidia (SEV2); and iii) severity of berry-to-berry infection caused by mycelium (SEV3). The model was validated in 21 epidemics (vineyard × year combinations) between 2009 and 2014 in Italy and France. A discriminant function analysis (DFA) was used to: i) evaluate the ability of the model to predict mild, intermediate, and severe epidemics; and ii) assess how SEV1, SEV2, and SEV3 contribute to epidemics. The model correctly classified the severity of 17 of 21 epidemics. Results from DFA were also used to calculate the daily probabilities that an ongoing epidemic would be mild, intermediate, or severe. SEV1 was the most influential variable in discriminating between mild and intermediate epidemics, whereas SEV2 and SEV3 were relevant for discriminating between intermediate and severe epidemics. The model represents an improvement of previous B. cinerea models in viticulture and could be useful for making decisions about Botrytis bunch rot control. PMID:26457808

  20. BcGs1, a glycoprotein from Botrytis cinerea, elicits defence response and improves disease resistance in host plants.

    PubMed

    Zhang, Yi; Zhang, Yunhua; Qiu, Dewen; Zeng, Hongmei; Guo, Lihua; Yang, Xiufen

    2015-02-20

    In this study, a necrosis-inducing protein was purified from the culture filtrate of the necrotrophic fungus Botrytis cinerea BC-98 strain. Secreted proteins were collected and fractionated by liquid chromatography. The fraction with the highest necrosis-inducing activity was further purified. A glycoprotein named BcGs1 was identified by 2D electrophoresis and mass spectrometry. The BcGs1 protein consisted of 672 amino acids with a theoretical molecular weight of 70.487 kDa. Functional domain analysis indicated that BcGs1 was a glucan 1,4-alpha-glucosidase, a cell wall-degrading enzyme, with a Glyco_hydro_15 domain and a CBM20_glucoamylase domain. The BcGs1 protein caused necrotic lesions that mimicked a typical hypersensitive response and H2O2 production in tomato and tobacco leaves. BcGs1-treated plants exhibited resistance to B. cinerea, Pseudomonas syringae pv. tomato DC3000 and tobacco mosaic virus in systemic leaves. In addition, BcGs1 triggered elevation of the transcript levels of the defence-related genes PR-1a, TPK1b and Prosystemin. This is the first report of a Botrytis glucan 1,4-alpha-glucosidase triggering host plant immunity as an elicitor. These results lay a foundation for further study of the comprehensive interaction between plants and necrotrophic fungi. PMID:25613865

  1. Negative regulation of ABA signaling by WRKY33 is critical for Arabidopsis immunity towards Botrytis cinerea 2100

    PubMed Central

    Liu, Shouan; Kracher, Barbara; Ziegler, Jörg; Birkenbihl, Rainer P; Somssich, Imre E

    2015-01-01

    The Arabidopsis mutant wrky33 is highly susceptible to Botrytis cinerea. We identified >1680 Botrytis-induced WRKY33 binding sites associated with 1576 Arabidopsis genes. Transcriptional profiling defined 318 functional direct target genes at 14 hr post inoculation. Comparative analyses revealed that WRKY33 possesses dual functionality acting either as a repressor or as an activator in a promoter-context dependent manner. We confirmed known WRKY33 targets involved in hormone signaling and phytoalexin biosynthesis, but also uncovered a novel negative role of abscisic acid (ABA) in resistance towards B. cinerea 2100. The ABA biosynthesis genes NCED3 and NCED5 were identified as direct targets required for WRKY33-mediated resistance. Loss-of-WRKY33 function resulted in elevated ABA levels and genetic studies confirmed that WRKY33 acts upstream of NCED3/NCED5 to negatively regulate ABA biosynthesis. This study provides the first detailed view of the genome-wide contribution of a specific plant transcription factor in modulating the transcriptional network associated with plant immunity. DOI: http://dx.doi.org/10.7554/eLife.07295.001 PMID:26076231

  2. Characterization of endophytic Bacillus strains from tomato plants (Lycopersicon esculentum) displaying antifungal activity against Botrytis cinerea Pers.

    PubMed

    Kefi, Asma; Ben Slimene, Imen; Karkouch, Ines; Rihouey, Christophe; Azaeiz, Sana; Bejaoui, Marwa; Belaid, Rania; Cosette, Pascal; Jouenne, Thierry; Limam, Ferid

    2015-12-01

    Eighty endophytic bacteria were isolated from healthy tissues of roots, stems, leaves and fruits of tomato plants (Lycopersicon esculentum). Four strains, named BL1, BT5, BR8 and BF11 were selected for their antagonism against Botrytis cinerea, a phytopathogenic fungus responsible of gray mold in several important crops, with growth inhibitory activity ranging from 27 to 53%. Morphological, biochemical, and molecular parameters as 16S rDNA sequencing demonstrated that the selected bacterial strains were related to Bacillus species which are known to produce and secrete a lot of lipopeptides with strong inhibitory effect against pathogen mycelial growth. Electrospray mass spectrometry analysis showed that these strains produced heterogeneous mixture of antibiotics belonging to fengycin and surfactin for BL1 and BT5, to iturin and surfactin for BR8, to bacillomycin D, fengycin and surfactin for BF11. Furthermore, these bacteria exhibited biocontrol potential by reducing the disease severity when tested on detached leaflets. Based on their antifungal activity against Botrytis cinerea, these strains could be used for biological control of plant diseases. PMID:26347324

  3. A Mechanistic Model of Botrytis cinerea on Grapevines That Includes Weather, Vine Growth Stage, and the Main Infection Pathways

    PubMed Central

    González-Domínguez, Elisa; Caffi, Tito; Ciliberti, Nicola; Rossi, Vittorio

    2015-01-01

    A mechanistic model for Botrytis cinerea on grapevine was developed. The model, which accounts for conidia production on various inoculum sources and for multiple infection pathways, considers two infection periods. During the first period (“inflorescences clearly visible” to “berries groat-sized”), the model calculates: i) infection severity on inflorescences and young clusters caused by conidia (SEV1). During the second period (“majority of berries touching” to “berries ripe for harvest”), the model calculates: ii) infection severity of ripening berries by conidia (SEV2); and iii) severity of berry-to-berry infection caused by mycelium (SEV3). The model was validated in 21 epidemics (vineyard × year combinations) between 2009 and 2014 in Italy and France. A discriminant function analysis (DFA) was used to: i) evaluate the ability of the model to predict mild, intermediate, and severe epidemics; and ii) assess how SEV1, SEV2, and SEV3 contribute to epidemics. The model correctly classified the severity of 17 of 21 epidemics. Results from DFA were also used to calculate the daily probabilities that an ongoing epidemic would be mild, intermediate, or severe. SEV1 was the most influential variable in discriminating between mild and intermediate epidemics, whereas SEV2 and SEV3 were relevant for discriminating between intermediate and severe epidemics. The model represents an improvement of previous B. cinerea models in viticulture and could be useful for making decisions about Botrytis bunch rot control. PMID:26457808

  4. Transcriptome analysis reveals genes commonly induced by Botrytis cinerea infection, cold, drought and oxidative stresses in Arabidopsis.

    PubMed

    Sham, Arjun; Al-Azzawi, Ahmed; Al-Ameri, Salma; Al-Mahmoud, Bassam; Awwad, Falah; Al-Rawashdeh, Ahmed; Iratni, Rabah; AbuQamar, Synan

    2014-01-01

    Signaling pathways controlling biotic and abiotic stress responses may interact synergistically or antagonistically. To identify the similarities and differences among responses to diverse stresses, we analyzed previously published microarray data on the transcriptomic responses of Arabidopsis to infection with Botrytis cinerea (a biotic stress), and to cold, drought, and oxidative stresses (abiotic stresses). Our analyses showed that at early stages after B. cinerea inoculation, 1498 genes were up-regulated (B. cinerea up-regulated genes; BUGs) and 1138 genes were down-regulated (B. cinerea down-regulated genes; BDGs). We showed a unique program of gene expression was activated in response each biotic and abiotic stress, but that some genes were similarly induced or repressed by all of the tested stresses. Of the identified BUGs, 25%, 6% and 12% were also induced by cold, drought and oxidative stress, respectively; whereas 33%, 7% and 5.5% of the BDGs were also down-regulated by the same abiotic stresses. Coexpression and protein-protein interaction network analyses revealed a dynamic range in the expression levels of genes encoding regulatory proteins. Analysis of gene expression in response to electrophilic oxylipins suggested that these compounds are involved in mediating responses to B. cinerea infection and abiotic stress through TGA transcription factors. Our results suggest an overlap among genes involved in the responses to biotic and abiotic stresses in Arabidopsis. Changes in the transcript levels of genes encoding components of the cyclopentenone signaling pathway in response to biotic and abiotic stresses suggest that the oxylipin signal transduction pathway plays a role in plant defense. Identifying genes that are commonly expressed in response to environmental stresses, and further analyzing the functions of their encoded products, will increase our understanding of the plant stress response. This information could identify targets for genetic

  5. Transcriptome Analysis Reveals Genes Commonly Induced by Botrytis cinerea Infection, Cold, Drought and Oxidative Stresses in Arabidopsis

    PubMed Central

    Al-Ameri, Salma; Al-Mahmoud, Bassam; Awwad, Falah; Al-Rawashdeh, Ahmed; Iratni, Rabah; AbuQamar, Synan

    2014-01-01

    Signaling pathways controlling biotic and abiotic stress responses may interact synergistically or antagonistically. To identify the similarities and differences among responses to diverse stresses, we analyzed previously published microarray data on the transcriptomic responses of Arabidopsis to infection with Botrytis cinerea (a biotic stress), and to cold, drought, and oxidative stresses (abiotic stresses). Our analyses showed that at early stages after B. cinerea inoculation, 1498 genes were up-regulated (B. cinerea up-regulated genes; BUGs) and 1138 genes were down-regulated (B. cinerea down-regulated genes; BDGs). We showed a unique program of gene expression was activated in response each biotic and abiotic stress, but that some genes were similarly induced or repressed by all of the tested stresses. Of the identified BUGs, 25%, 6% and 12% were also induced by cold, drought and oxidative stress, respectively; whereas 33%, 7% and 5.5% of the BDGs were also down-regulated by the same abiotic stresses. Coexpression and protein-protein interaction network analyses revealed a dynamic range in the expression levels of genes encoding regulatory proteins. Analysis of gene expression in response to electrophilic oxylipins suggested that these compounds are involved in mediating responses to B. cinerea infection and abiotic stress through TGA transcription factors. Our results suggest an overlap among genes involved in the responses to biotic and abiotic stresses in Arabidopsis. Changes in the transcript levels of genes encoding components of the cyclopentenone signaling pathway in response to biotic and abiotic stresses suggest that the oxylipin signal transduction pathway plays a role in plant defense. Identifying genes that are commonly expressed in response to environmental stresses, and further analyzing the functions of their encoded products, will increase our understanding of the plant stress response. This information could identify targets for genetic

  6. Effect of cuticular waxes compounds from table grapes on growth, germination and gene expression in Botrytis cinerea.

    PubMed

    Silva-Moreno, Evelyn; Brito-Echeverría, Jocelyn; López, Miguel; Ríos, Juan; Balic, Iván; Campos-Vargas, Reinaldo; Polanco, Rubén

    2016-05-01

    Botrytis cinerea attacks a broad range of host causing significant economic losses in the worldwide fruit export industry. Hitherto, many studies have focused on the penetration mechanisms used by this phytopathogen, but little is known about the early stages of infection, especially those such as adhesion and germination. The aim of this work was to evaluate the effect of cuticular waxes compounds from table grapes on growth, germination and gene expression of B. cinerea. To accomplish this, growth was analyzed using as substrate n-alkanes extracted from waxes of fresh fruit (table grapes, blueberries and apricots). Subsequently, the main compounds of table grape waxes, oleanolic acid (OA) and n-fatty alcohols, were mixed to generate a matrix on which conidia of B. cinerea were added to assess their effect on germination and expression of bctub, bchtr and bchex genes. B. cinerea B05.10, isolated from grapes, increased its growth on a matrix composed by table grapes n-alkanes in comparison to a matrix made with n-alkanes from apricot or blueberries. Moreover, at 2.5 h, B05.10 germination increased 17 and 33 % in presence of n-alkanes from table grape, in comparison to conditions without alkanes or with blueberries alkanes, respectively. Finally, expression of bchtr and bchex showed a significant increase during the first hour after contact with n-fatty alcohols and OA. In conclusion, B. cinerea displays selectivity towards certain compounds found in host waxes, mainly n-fatty alcohols, which could be a good candidate to control this phytopathogen in early stages of infection. PMID:27038944

  7. Reactive oxygen and nitrogen (ROS and RNS) species generation and cell death in tomato suspension cultures--Botrytis cinerea interaction.

    PubMed

    Pietrowska, E; Różalska, S; Kaźmierczak, A; Nawrocka, J; Małolepsza, U

    2015-01-01

    This article reports events connected to cell survival and Botrytis cinerea infection development in cell suspension cultures of two tomato cultivars which show different levels of susceptibility to the pathogen: cv. Corindo (more susceptible) and cv. Perkoz (less susceptible). In parallel changes in reactive oxygen (ROS) and nitrogen (RNS) species generation and in S-nitrosoglutathione reductase (GSNOR) activity were studied. In vivo staining methods with acridine orange (AO) and ethidium bromide (EB) as well as fluorescent microscopy were used to assess tomato and B. cinerea cells death. The biochemical studies of ROS and RNS concentrations in plant cell extract were complemented by in vivo ROS and nitric oxide (NO) imaging using nitro blue tetrazolium (NBT), diaminobenzidine (DAB) and diaminofluorescein diacetate (DAF-DA) staining methods, and confocal microscope technique. B. cinerea infection proceeded slower in Perkoz cell cultures. It was evidenced by measuring the pathogen conidia germination and germination tube development in which nuclei revealing cell death dominated. Two different types of tomato cell death were observed: cells with necrotic nuclei dominated in Corindo whereas in Perkoz cells with characteristic of vacuolar death type prevailed. In Perkoz cells, constitutive levels of NO and S-nitrosothiols (SNO) were significantly higher and hydrogen peroxide (H₂O₂) and superoxide anion (O₂(-)) concentrations were slightly higher as compared with Corindo cells. Moreover, increases in these molecule concentrations as a result of B. cinerea inoculation were observed in both, Perkoz and Corindo cell cultures. The enzymatic GSNOR activity seems to be an important player in controlling the SNO level in tomato cells. Involvements of the studied compounds in molecular mechanisms of tomato resistance to B. cinerea are discussed in the paper. PMID:25064634

  8. Arabidopsis AtERF15 positively regulates immunity against Pseudomonas syringae pv. tomato DC3000 and Botrytis cinerea

    PubMed Central

    Zhang, Huijuan; Huang, Lei; Dai, Yi; Liu, Shixia; Hong, Yongbo; Tian, Limei; Huang, Lihong; Cao, Zhongye; Li, Dayong; Song, Fengming

    2015-01-01

    Upon pathogen infection, activation of immune response requires effective transcriptional reprogramming that regulates inducible expression of a large set of defense genes. A number of ethylene-responsive factor transcription factors have been shown to play critical roles in regulating immune responses in plants. In the present study, we explored the functions of Arabidopsis AtERF15 in immune responses against Pseudomonas syringae pv. tomato (Pst) DC3000, a (hemi)biotrophic bacterial pathogen, and Botrytis cinerea, a necrotrophic fungal pathogen. Expression of AtERF15 was induced by infection of Pst DC3000 and B. cinerea and by treatments with salicylic acid (SA) and methyl jasmonate. Biochemical assays demonstrated that AtERF15 is a nucleus-localized transcription activator. The AtERF15-overexpressing (AtERF15-OE) plants displayed enhanced resistance while the AtERF15-RNAi plants exhibited decreased resistance against Pst DC3000 and B. cinerea. Meanwhile, Pst DC3000- or B. cinerea-induced expression of defense genes was upregulated in AtERF15-OE plants but downregulated in AtERF15-RNAi plants, as compared to the expression in wild type plants. In response to infection with B. cinerea, the AtERF15-OE plants accumulated less reactive oxygen species (ROS) while the AtERF15-RNAi plants accumulated more ROS. The flg22- and chitin-induced oxidative burst was abolished and expression levels of the pattern-triggered immunity-responsive genes AtFRK1 and AtWRKY53 were suppressed in AtER15-RNAi plants upon treatment with flg22 or chitin. Furthermore, SA-induced defense response was also partially impaired in the AtERF15-RNAi plants. These data demonstrate that AtERF15 is a positive regulator of multiple layers of the immune responses in Arabidopsis. PMID:26388886

  9. Effect of gamma irradiation and its convergent treatment for control of postharvest Botrytis cinerea of cut roses

    NASA Astrophysics Data System (ADS)

    Chu, Eun-Hee; Shin, Eun-Jung; Park, Hae-Jun; Jeong, Rae-Dong

    2015-10-01

    Postharvest diseases cause considerable losses to harvested crops. Among them, gray mold (Botrytis cinerea) is a major problem of exporting to cut rose flowers into Korea. Irradiation treatment is an alternative to phytosanitary purposes and a useful nonchemical approach to the control of postharvest diseases. Gamma irradiation was evaluated for its in vitro and in vivo antifungal activity against B. cinerea on cut rose varieties, 'Shooting Star' and 'Babe'. The irradiating dose required to reduce the population by 90%, D10, was 0.99 kGy. Gamma irradiation showed complete inhibition of spore germination and mycelial growth of B. cinerea, especially 4.0 kGy in vitro. Antifungal activity of gamma irradiation on rose B. cinerea is a dose-dependent manner. A significant phytotoxicity such as bent neck in cut rose quality was shown from gamma irradiation at over 0.4 kGy (p<0.05) in both varieties. Although there is no significant difference in both varieties for fresh weight, in the case of flower rate, 'Babe' shows more sensitivity than 'Shooting Star'. In vivo assays demonstrated that established doses in in vitro, over 4 kGy, could completely inactive fungal pathogens, but such high doses can cause severe flowers damage. Thus, to eliminate negative impact on their quality, gamma irradiation was evaluated at lower doses in combination with an eco-friendly chemical, sodium dichloroisocyanurate (NaDCC) to examine the inhibition of B. cinerea. Intriguingly, only the combined treatment with 0.2 kGy of gamma irradiation and 70 ppm of NaDCC exhibited significant synergistic antifungal activity against blue mold decay in both varieties. Together, these results suggest that a synergistic effect of the combined treatment with gamma irradiation and NaDCC can be efficiently used to control the postharvest diseases in cut rose flowers, and will provide a promising technology for horticulture products for exportation.

  10. Toxicity of ozone gas to conidia of Penicillium digitatum, P. italicum, and Botrytis cinerea and control of gray mold on table grapes.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium digitatum, P. italicum, and Botrytis cinerea attack fresh fruit and cause significant postharvest decay losses and the toxicity of ozone (O3) gas at different relative humidities to control their conidia was determined. Conidia were exposed to an atmosphere containing 200 to 350 µL L-1 o...

  11. Metabolomic approaches reveal that cell wall modifications play a major role in ethylene-mediated resistance against Botrytis cinerea.

    PubMed

    Lloyd, Amanda J; William Allwood, J; Winder, Catherine L; Dunn, Warwick B; Heald, James K; Cristescu, Simona M; Sivakumaran, Anushen; Harren, Frans J M; Mulema, Joseph; Denby, Katherine; Goodacre, Royston; Smith, Aileen R; Mur, Luis A J

    2011-09-01

    In Arabidopsis, resistance to the necrotrophic fungus Botrytis cinerea is conferred by ethylene via poorly understood mechanisms. Metabolomic approaches compared the responses of the wild-type, the ethylene-insensitive mutant etr1-1, which showed increased susceptibility, and the constitutively active ethylene mutants ctr1-1 and eto2 both exhibited decreased susceptibility to B. cinerea. Fourier transform-infrared (FT-IR) spectroscopy demonstrated reproducible biochemical differences between treatments and genotypes. To identify discriminatory mass-to-charge ratios (m/z) associated with resistance, discriminant function analysis was employed on spectra derived from direct injection electrospray ionisation-mass spectrometry on the derived principal components of these data. Ethylene-modulated m/z were mapped onto Arabidopsis biochemical pathways and many were associated with hydroxycinnamate and monolignol biosynthesis, both linked to cell wall modification. A high-resolution linear triple quadrupole-Orbitrap hybrid system confirmed the identity of key metabolites in these pathways. The contribution of these pathways to defence against B. cinerea was validated through the use of multiple Arabidopsis mutants. The FT-IR microspectroscopy indicated that spatial accumulation of hydroxycinnamates and monolignols at the cell wall to confine disease was linked ot ethylene. These data demonstrate the power of metabolomic approaches in elucidating novel biological phenomena, especially when coupled to validation steps exploiting relevant mutant genotypes. PMID:21575089

  12. Transcriptome analysis reveals regulatory networks underlying differential susceptibility to Botrytis cinerea in response to nitrogen availability in Solanum lycopersicum

    PubMed Central

    Vega, Andrea; Canessa, Paulo; Hoppe, Gustavo; Retamal, Ignacio; Moyano, Tomas C.; Canales, Javier; Gutiérrez, Rodrigo A.; Rubilar, Joselyn

    2015-01-01

    Nitrogen (N) is one of the main limiting nutrients for plant growth and crop yield. It is well documented that changes in nitrate availability, the main N source found in agricultural soils, influences a myriad of developmental programs and processes including the plant defense response. Indeed, many agronomical reports indicate that the plant N nutritional status influences their ability to respond effectively when challenged by different pathogens. However, the molecular mechanisms involved in N-modulation of plant susceptibility to pathogens are poorly characterized. In this work, we show that Solanum lycopersicum defense response to the necrotrophic fungus Botrytis cinerea is affected by plant N availability, with higher susceptibility in nitrate-limiting conditions. Global gene expression responses of tomato against B. cinerea under contrasting nitrate conditions reveals that plant primary metabolism is affected by the fungal infection regardless of N regimes. This result suggests that differential susceptibility to pathogen attack under contrasting N conditions is not only explained by a metabolic alteration. We used a systems biology approach to identify the transcriptional regulatory network implicated in plant response to the fungus infection under contrasting nitrate conditions. Interestingly, hub genes in this network are known key transcription factors involved in ethylene and jasmonic acid signaling. This result positions these hormones as key integrators of nitrate and defense against B. cinerea in tomato plants. Our results provide insights into potential crosstalk mechanisms between necrotrophic defense response and N status in plants. PMID:26583019

  13. The ‘Dr Jekyll and Mr Hyde fungus’: noble rot versus gray mold symptoms of Botrytis cinerea on grapes

    PubMed Central

    Fournier, Elisabeth; Gladieux, Pierre; Giraud, Tatiana

    2013-01-01

    Many cryptic species have recently been discovered in fungi, especially in fungal plant pathogens. Cryptic fungal species co-occurring in sympatry may occupy slightly different ecological niches, for example infecting the same crop plant but specialized on different organs or having different phenologies. Identifying cryptic species in fungal pathogens of crops and determining their ecological specialization are therefore crucial for disease management. Here, we addressed this question in the ascomycete Botrytis cinerea, the agent of gray mold on a wide range of plants. On grape, B. cinerea causes severe damage but is also responsible for noble rot used for processing sweet wines. We used microsatellite genotyping and clustering methods to elucidate whether isolates sampled on gray mold versus noble rot symptoms in three French regions belong to genetically differentiated populations. The inferred population structure matched geography rather than the type of symptom. Noble rot symptoms therefore do not seem to be caused by a specific B. cinerea population but instead seem to depend essentially on microclimatic conditions, which has applied consequences for the production of sweet wines. PMID:24062804

  14. Arabidopsis Defense against Botrytis cinerea: Chronology and Regulation Deciphered by High-Resolution Temporal Transcriptomic Analysis[C][W

    PubMed Central

    Windram, Oliver; Madhou, Priyadharshini; McHattie, Stuart; Hill, Claire; Hickman, Richard; Cooke, Emma; Jenkins, Dafyd J.; Penfold, Christopher A.; Baxter, Laura; Breeze, Emily; Kiddle, Steven J.; Rhodes, Johanna; Atwell, Susanna; Kliebenstein, Daniel J.; Kim, Youn-sung; Stegle, Oliver; Borgwardt, Karsten; Zhang, Cunjin; Tabrett, Alex; Legaie, Roxane; Moore, Jonathan; Finkenstadt, Bärbel; Wild, David L.; Mead, Andrew; Rand, David; Beynon, Jim; Ott, Sascha; Buchanan-Wollaston, Vicky; Denby, Katherine J.

    2012-01-01

    Transcriptional reprogramming forms a major part of a plant’s response to pathogen infection. Many individual components and pathways operating during plant defense have been identified, but our knowledge of how these different components interact is still rudimentary. We generated a high-resolution time series of gene expression profiles from a single Arabidopsis thaliana leaf during infection by the necrotrophic fungal pathogen Botrytis cinerea. Approximately one-third of the Arabidopsis genome is differentially expressed during the first 48 h after infection, with the majority of changes in gene expression occurring before significant lesion development. We used computational tools to obtain a detailed chronology of the defense response against B. cinerea, highlighting the times at which signaling and metabolic processes change, and identify transcription factor families operating at different times after infection. Motif enrichment and network inference predicted regulatory interactions, and testing of one such prediction identified a role for TGA3 in defense against necrotrophic pathogens. These data provide an unprecedented level of detail about transcriptional changes during a defense response and are suited to systems biology analyses to generate predictive models of the gene regulatory networks mediating the Arabidopsis response to B. cinerea. PMID:23023172

  15. Histone H3 Lysine 9 Methyltransferase DIM5 Is Required for the Development and Virulence of Botrytis cinerea

    PubMed Central

    Zhang, Xiaoli; Liu, Xinqiang; Zhao, Yanli; Cheng, Jiasen; Xie, Jiatao; Fu, Yanping; Jiang, Daohong; Chen, Tao

    2016-01-01

    Histone methylation is widely present in animals, plants and fungi, and the methylation modification of histone H3 has important biological functions. Methylation of Lys9 of histone H3 (H3K9) has been proven to regulate chromatin structure, gene silencing, transcriptional activation, plant metabolism, and other processes. In this work, we investigated the functions of a H3K9 methyltransferase gene BcDIM5 in Botrytis cinerea, which contains a PreSET domain, a SET domain and a PostSET domain. Characterization of BcDIM5 knockout transformants showed that the hyphal growth rate and production of conidiophores and sclerotia were significantly reduced, while complementary transformation of BcDIM5 could restore the phenotypes to the levels of wild type. Pathogenicity assays revealed that BcDIM5 was essential for full virulence of B. cinerea. BcDIM5 knockout transformants exhibited decreased virulence, down-regulated expression of some pathogenic genes and drastically decreased H3K9 trimethylation level. However, knockout transformants of other two genes heterochromatin protein 1 (HP1) BcHP1 and DNA methyltransferase (DIM2) BcDIM2 did not exhibit significant change in the growth phenotype and virulence compared with the wild type. Our results indicate that H3K9 methyltransferase BcDIM5 is required for H3K9 trimethylation to regulate the development and virulence of B. cinerea. PMID:27597848

  16. Vitamin B6 contributes to disease resistance against Pseudomonas syringae pv. tomato DC3000 and Botrytis cinerea in Arabidopsis thaliana.

    PubMed

    Zhang, Yafen; Jin, Xiaoyi; Ouyang, Zhigang; Li, Xiaohui; Liu, Bo; Huang, Lei; Hong, Yongbo; Zhang, Huijuan; Song, Fengming; Li, Dayong

    2015-03-01

    Vitamin B6 (VB6) is an important cofactor for numerous enzymatic reactions and plays an important role in abiotic stress tolerance. However, direct molecular evidence supporting a role for VB6 in plant disease resistance remains lacking. In this study, we explored the possible function of VB6 in disease resistance by analyzing disease phenotypes of Arabidopsis mutants with defects in de novo biosynthetic pathway and salvage pathway of VB6 biosynthesis against Pseudomonas syringae pv. tomato (Pst) DC3000 and Botrytis cinerea. Mutations in AtPDX1.2 and AtPDX1.3 genes involved in the de novo pathway, and in AtSOS4 gene involved in the salvage pathway led to increased levels of diseases caused by Pst DC3000 and B. cinerea. The pdx1.2 and pdx1.3 plants had reduced VB6 contents and showed a further reduction in VB6 contents after infection by Pst DC3000 or B. cinerea. Our preliminary results suggest an important role for VB6 in plant disease resistance against different types of pathogens. PMID:25460872

  17. Transcriptome analysis reveals regulatory networks underlying differential susceptibility to Botrytis cinerea in response to nitrogen availability in Solanum lycopersicum.

    PubMed

    Vega, Andrea; Canessa, Paulo; Hoppe, Gustavo; Retamal, Ignacio; Moyano, Tomas C; Canales, Javier; Gutiérrez, Rodrigo A; Rubilar, Joselyn

    2015-01-01

    Nitrogen (N) is one of the main limiting nutrients for plant growth and crop yield. It is well documented that changes in nitrate availability, the main N source found in agricultural soils, influences a myriad of developmental programs and processes including the plant defense response. Indeed, many agronomical reports indicate that the plant N nutritional status influences their ability to respond effectively when challenged by different pathogens. However, the molecular mechanisms involved in N-modulation of plant susceptibility to pathogens are poorly characterized. In this work, we show that Solanum lycopersicum defense response to the necrotrophic fungus Botrytis cinerea is affected by plant N availability, with higher susceptibility in nitrate-limiting conditions. Global gene expression responses of tomato against B. cinerea under contrasting nitrate conditions reveals that plant primary metabolism is affected by the fungal infection regardless of N regimes. This result suggests that differential susceptibility to pathogen attack under contrasting N conditions is not only explained by a metabolic alteration. We used a systems biology approach to identify the transcriptional regulatory network implicated in plant response to the fungus infection under contrasting nitrate conditions. Interestingly, hub genes in this network are known key transcription factors involved in ethylene and jasmonic acid signaling. This result positions these hormones as key integrators of nitrate and defense against B. cinerea in tomato plants. Our results provide insights into potential crosstalk mechanisms between necrotrophic defense response and N status in plants. PMID:26583019

  18. Features of air masses associated with the deposition of Pseudomonas syringae and Botrytis cinerea by rain and snowfall

    PubMed Central

    Monteil, Caroline L; Bardin, Marc; Morris, Cindy E

    2014-01-01

    Clarifying the role of precipitation in microbial dissemination is essential for elucidating the processes involved in disease emergence and spread. The ecology of Pseudomonas syringae and its presence throughout the water cycle makes it an excellent model to address this issue. In this study, 90 samples of freshly fallen rain and snow collected from 2005–2011 in France were analyzed for microbiological composition. The conditions favorable for dissemination of P. syringae by this precipitation were investigated by (i) estimating the physical properties and backward trajectories of the air masses associated with each precipitation event and by (ii) characterizing precipitation chemistry, and genetic and phenotypic structures of populations. A parallel study with the fungus Botrytis cinerea was also performed for comparison. Results showed that (i) the relationship of P. syringae to precipitation as a dissemination vector is not the same for snowfall and rainfall, whereas it is the same for B. cinerea and (ii) the occurrence of P. syringae in precipitation can be linked to electrical conductivity and pH of water, the trajectory of the air mass associated with the precipitation and certain physical conditions of the air mass (i.e. temperature, solar radiation exposure, distance traveled), whereas these predictions are different for B. cinerea. These results are pertinent to understanding microbial survival, emission sources and atmospheric processes and how they influence microbial dissemination. PMID:24722630

  19. Histone H3 Lysine 9 Methyltransferase DIM5 Is Required for the Development and Virulence of Botrytis cinerea.

    PubMed

    Zhang, Xiaoli; Liu, Xinqiang; Zhao, Yanli; Cheng, Jiasen; Xie, Jiatao; Fu, Yanping; Jiang, Daohong; Chen, Tao

    2016-01-01

    Histone methylation is widely present in animals, plants and fungi, and the methylation modification of histone H3 has important biological functions. Methylation of Lys9 of histone H3 (H3K9) has been proven to regulate chromatin structure, gene silencing, transcriptional activation, plant metabolism, and other processes. In this work, we investigated the functions of a H3K9 methyltransferase gene BcDIM5 in Botrytis cinerea, which contains a PreSET domain, a SET domain and a PostSET domain. Characterization of BcDIM5 knockout transformants showed that the hyphal growth rate and production of conidiophores and sclerotia were significantly reduced, while complementary transformation of BcDIM5 could restore the phenotypes to the levels of wild type. Pathogenicity assays revealed that BcDIM5 was essential for full virulence of B. cinerea. BcDIM5 knockout transformants exhibited decreased virulence, down-regulated expression of some pathogenic genes and drastically decreased H3K9 trimethylation level. However, knockout transformants of other two genes heterochromatin protein 1 (HP1) BcHP1 and DNA methyltransferase (DIM2) BcDIM2 did not exhibit significant change in the growth phenotype and virulence compared with the wild type. Our results indicate that H3K9 methyltransferase BcDIM5 is required for H3K9 trimethylation to regulate the development and virulence of B. cinerea. PMID:27597848

  20. Botrytis pseudocinerea Is a Significant Pathogen of Several Crop Plants but Susceptible to Displacement by Fungicide-Resistant B. cinerea Strains.

    PubMed

    Plesken, Cecilia; Weber, Roland W S; Rupp, Sabrina; Leroch, Michaela; Hahn, Matthias

    2015-10-01

    Botrytis cinerea is one of the most important pathogens worldwide, causing gray mold on a large variety of crops. Botrytis pseudocinerea has been found previously to occur together with B. cinerea in low abundance in vineyards and strawberry fields. Here, we report B. pseudocinerea to be common and sometimes dominant over B. cinerea on several fruit and vegetable crops in Germany. On apples with calyx end rot and on oilseed rape, it was the major gray mold species. Abundance of B. pseudocinerea was often negatively correlated with fungicide treatments. On cultivated strawberries, it was frequently found in spring but was largely displaced by B. cinerea following fungicide applications. Whereas B. cinerea strains with multiple-fungicide resistance were common in these fields, B. pseudocinerea almost never developed resistance to any fungicide even though resistance mutations occurred at similar frequencies in both species under laboratory conditions. The absence of resistance to quinone outside inhibitors in B. pseudocinerea was correlated with an intron in cytB preventing the major G143A resistance mutation. Our work indicates that B. pseudocinerea has a wide host range similar to that of B. cinerea and that it can become an important gray mold pathogen on cultivated plants. PMID:26231644

  1. Botrytis pseudocinerea Is a Significant Pathogen of Several Crop Plants but Susceptible to Displacement by Fungicide-Resistant B. cinerea Strains

    PubMed Central

    Plesken, Cecilia; Weber, Roland W. S.; Rupp, Sabrina; Leroch, Michaela

    2015-01-01

    Botrytis cinerea is one of the most important pathogens worldwide, causing gray mold on a large variety of crops. Botrytis pseudocinerea has been found previously to occur together with B. cinerea in low abundance in vineyards and strawberry fields. Here, we report B. pseudocinerea to be common and sometimes dominant over B. cinerea on several fruit and vegetable crops in Germany. On apples with calyx end rot and on oilseed rape, it was the major gray mold species. Abundance of B. pseudocinerea was often negatively correlated with fungicide treatments. On cultivated strawberries, it was frequently found in spring but was largely displaced by B. cinerea following fungicide applications. Whereas B. cinerea strains with multiple-fungicide resistance were common in these fields, B. pseudocinerea almost never developed resistance to any fungicide even though resistance mutations occurred at similar frequencies in both species under laboratory conditions. The absence of resistance to quinone outside inhibitors in B. pseudocinerea was correlated with an intron in cytB preventing the major G143A resistance mutation. Our work indicates that B. pseudocinerea has a wide host range similar to that of B. cinerea and that it can become an important gray mold pathogen on cultivated plants. PMID:26231644

  2. Effects of Botrytis cinerea and Pseudomonas syringae infection on the antioxidant profile of Mesembryanthemum crystallinum C3/CAM intermediate plant.

    PubMed

    Libik-Konieczny, Marta; Surówka, Ewa; Kuźniak, Elżbieta; Nosek, Michał; Miszalski, Zbigniew

    2011-07-01

    Mesembryathemum crystallinum plants performing C(3) or CAM (crassulacean acid metabolism) appear to be highly resistant to Botrytis cinerea as well as to Pseudomonas syringae. Fungal hyphae growth was restricted to 48h post-inoculation (hpi) in both metabolic types and morphology of hyphae differed between those growing in C(3) and CAM plants. Growth of bacteria was inhibited significantly 24 hpi in both C(3) and CAM plants. B. cinerea and P. syringae infection led to an increase in the concentration of H(2)O(2) in C(3) plants 3 hpi, while a decrease in H(2)O(2) content was observed in CAM performing plants. The concentration of H(2)O(2) returned to the control level 24 and 48 hpi. Changes in H(2)O(2) content corresponded with the activity of guaiacol peroxidase (POD), mostly 3 hpi. We noted that its activity decreased significantly in C(3) plants and increased in CAM plants in response to inoculation with both pathogens. On the contrary, changes in the activity of CAT did not correlate with H(2)O(2) level. It increased significantly after interaction of C(3) plants with B. cinerea or P. syringae, but in CAM performing plants, the activity of this enzyme was unchanged. Inoculation with B. cinerea or P. syringae led to an increase in the total SOD activity in C(3) plants while CAM plants did not exhibit changes in the total SOD activity after interaction with both pathogens. In conclusion, the pathogen-induced changes in H(2)O(2) content and in SOD, POD and CAT activities in M. crystallinum leaves, were related to the photosynthetic metabolism type of the stressed plants rather than to the lifestyle of the invading pathogen. PMID:21342714

  3. Effects of Airflow and Changing Humidity on the Aerosolization of Respirable Fungal Fragments and Conidia of Botrytis cinerea

    PubMed Central

    2012-01-01

    The purpose of this study was to investigate the aerosolization of particles (micro- and macroconidia and fragments) from Botrytis cinerea cultures in relation to potential human inhalation in indoor environments. The influence of the following factors on the aerosolization of B. cinerea particles was studied: exposure to airflow, relative humidity (rh), changing rh, and plant or building materials. The aerodynamic diameter (da) and the respirable fraction of the aerosolized particles were determined. Conidia and fragments of B. cinerea were not aerosolized as a response to a decrease in the rh. In contrast, both micro- and macroconidia and fungal fragments were aerosolized when exposed to an airflow of 1.5 m s−1 or 0.5 m s−1. Significantly more particles of microconidial size and fragment size were aerosolized at a low rh (18 to 40% rh) than at a higher rh (60 to 80% rh) when cultures were exposed to airflow. The size of the respirable fraction of the aerosolized particles was dependent on the rh but not on the growth material. At high rh, about 30% of the aerosolized particles were of respirable size, while at low rh, about 70% were of respirable size. During low rh, more fungal (1→3)-β-d-glucan and chitinase were aerosolized than during high rh. In conclusion, exposure to external physical forces such as airflow is necessary for the aerosolization of particles from B. cinerea. The amount and size distribution are highly affected by the rh, and more particles of respirable sizes were aerosolized at low rh than at high rh. PMID:22447608

  4. Unraveling the Function of the Response Regulator BcSkn7 in the Stress Signaling Network of Botrytis cinerea

    PubMed Central

    Viefhues, Anne; Schlathoelter, Ina; Simon, Adeline; Viaud, Muriel

    2015-01-01

    Important for the lifestyle and survival of every organism is the ability to respond to changing environmental conditions. The necrotrophic plant pathogen Botrytis cinerea triggers an oxidative burst in the course of plant infection and therefore needs efficient signal transduction to cope with this stress. The factors involved in this process and their precise roles are still not well known. Here, we show that the transcription factor Bap1 and the response regulator (RR) B. cinerea Skn7 (BcSkn7) are two key players in the oxidative stress response (OSR) of B. cinerea; both have a major influence on the regulation of classical OSR genes. A yeast-one-hybrid (Y1H) approach proved direct binding to the promoters of gsh1 and grx1 by Bap1 and of glr1 by BcSkn7. While the function of Bap1 is restricted to the regulation of oxidative stress, analyses of Δbcskn7 mutants revealed functions beyond the OSR. Involvement of BcSkn7 in development and virulence could be demonstrated, indicated by reduced vegetative growth, impaired formation of reproductive structures, and reduced infection cushion-mediated penetration of the host by the mutants. Furthermore, Δbcskn7 mutants were highly sensitive to oxidative, osmotic, and cell wall stress. Analyses of Δbap1 bcskn7 double mutants indicated that loss of BcSkn7 uncovers an underlying phenotype of Bap1. In contrast to Saccharomyces cerevisiae, the ortholog of the glutathione peroxidase Gpx3p is not required for nuclear translocation of Bap1. The presented results contribute to the understanding of the OSR in B. cinerea and prove that it differs substantially from that of yeast, demonstrating the complexity and versatility of components involved in signaling pathways. PMID:25934690

  5. Unraveling the Function of the Response Regulator BcSkn7 in the Stress Signaling Network of Botrytis cinerea.

    PubMed

    Viefhues, Anne; Schlathoelter, Ina; Simon, Adeline; Viaud, Muriel; Tudzynski, Paul

    2015-07-01

    Important for the lifestyle and survival of every organism is the ability to respond to changing environmental conditions. The necrotrophic plant pathogen Botrytis cinerea triggers an oxidative burst in the course of plant infection and therefore needs efficient signal transduction to cope with this stress. The factors involved in this process and their precise roles are still not well known. Here, we show that the transcription factor Bap1 and the response regulator (RR) B. cinerea Skn7 (BcSkn7) are two key players in the oxidative stress response (OSR) of B. cinerea; both have a major influence on the regulation of classical OSR genes. A yeast-one-hybrid (Y1H) approach proved direct binding to the promoters of gsh1 and grx1 by Bap1 and of glr1 by BcSkn7. While the function of Bap1 is restricted to the regulation of oxidative stress, analyses of Δbcskn7 mutants revealed functions beyond the OSR. Involvement of BcSkn7 in development and virulence could be demonstrated, indicated by reduced vegetative growth, impaired formation of reproductive structures, and reduced infection cushion-mediated penetration of the host by the mutants. Furthermore, Δbcskn7 mutants were highly sensitive to oxidative, osmotic, and cell wall stress. Analyses of Δbap1 bcskn7 double mutants indicated that loss of BcSkn7 uncovers an underlying phenotype of Bap1. In contrast to Saccharomyces cerevisiae, the ortholog of the glutathione peroxidase Gpx3p is not required for nuclear translocation of Bap1. The presented results contribute to the understanding of the OSR in B. cinerea and prove that it differs substantially from that of yeast, demonstrating the complexity and versatility of components involved in signaling pathways. PMID:25934690

  6. Requirement of Functional Ethylene-Insensitive 2 Gene for Efficient Resistance of Arabidopsis to Infection by Botrytis cinerea1

    PubMed Central

    Thomma, Bart P.H.J.; Eggermont, Kristel; Tierens, Koenraad F.M.-J.; Broekaert, Willem F.

    1999-01-01

    Inoculation of wild-type Arabidopsis plants with the fungus Alternaria brassicicola results in systemic induction of genes encoding a plant defensin (PDF1.2), a basic chitinase (PR-3), and an acidic hevein-like protein (PR-4). Pathogen-induced induction of these three genes is almost completely abolished in the ethylene-insensitive Arabidopsis mutant ein2-1. This indicates that a functional ethylene signal transduction component (EIN2) is required in this response. The ein2-1 mutants were found to be markedly more susceptible than wild-type plants to infection by two different strains of the gray mold fungus Botrytis cinerea. In contrast, no increased fungal colonization of ein2-1 mutants was observed after challenge with avirulent strains of either Peronospora parasitica or A. brassicicola. Our data support the conclusion that ethylene-controlled responses play a role in resistance of Arabidopsis to some but not all types of pathogens. PMID:10594097

  7. Beyond plant defense: insights on the potential of salicylic and methylsalicylic acid to contain growth of the phytopathogen Botrytis cinerea.

    PubMed

    Dieryckx, Cindy; Gaudin, Vanessa; Dupuy, Jean-William; Bonneu, Marc; Girard, Vincent; Job, Dominique

    2015-01-01

    Using Botrytis cinerea we confirmed in the present work several previous studies showing that salicylic acid, a main plant hormone, inhibits fungal growth in vitro. Such an inhibitory effect was also observed for the two salicylic acid derivatives, methylsalicylic and acetylsalicylic acid. In marked contrast, 5-sulfosalicylic acid was totally inactive. Comparative proteomics from treated vs. control mycelia showed that both the intracellular and extracellular proteomes were affected in the presence of salicylic acid or methylsalicylic acid. These data suggest several mechanisms that could potentially account for the observed fungal growth inhibition, notably pH regulation, metal homeostasis, mitochondrial respiration, ROS accumulation and cell wall remodeling. The present observations support a role played by the phytohormone SA and derivatives in directly containing the pathogen. Data are available via ProteomeXchange with identifier PXD002873. PMID:26528317

  8. Inhibitory effect of Xenorhabdus nematophila TB on plant pathogens Phytophthora capsici and Botrytis cinerea in vitro and in planta

    PubMed Central

    Fang, Xiangling; Zhang, Manrang; Tang, Qian; Wang, Yonghong; Zhang, Xing

    2014-01-01

    Entomopathogenic bacteria Xenorhabdus spp. produce secondary metabolites with potential antimicrobial activity for use in agricultural productions. This study evaluated the inhibitory effect of X. nematophila TB culture on plant pathogens Botrytis cinerea and Phytophthora capsici. The cell-free filtrate of TB culture showed strong inhibitory effects (>90%) on mycelial growth of both pathogens. The methanol-extracted bioactive compounds (methanol extract) of TB culture also had strong inhibitory effects on mycelial growth and spore germinations of both pathogens. The methanol extract (1000 μg/mL) and cell-free filtrate both showed strong therapeutic and protective effects (>70%) on grey mold both in detached tomato fruits and plants, and leaf scorch in pepper plants. This study demonstrates X. nematophila TB produces antimicrobial metabolites of strong activity on plant pathogens, with great potential for controlling tomato grey mold and pepper leaf scorch and being used in integrated disease control to reduce chemical application. PMID:24599183

  9. Inhibitory effect of Xenorhabdus nematophila TB on plant pathogens Phytophthora capsici and Botrytis cinerea in vitro and in planta.

    PubMed

    Fang, Xiangling; Zhang, Manrang; Tang, Qian; Wang, Yonghong; Zhang, Xing

    2014-01-01

    Entomopathogenic bacteria Xenorhabdus spp. produce secondary metabolites with potential antimicrobial activity for use in agricultural productions. This study evaluated the inhibitory effect of X. nematophila TB culture on plant pathogens Botrytis cinerea and Phytophthora capsici. The cell-free filtrate of TB culture showed strong inhibitory effects (>90%) on mycelial growth of both pathogens. The methanol-extracted bioactive compounds (methanol extract) of TB culture also had strong inhibitory effects on mycelial growth and spore germinations of both pathogens. The methanol extract (1000 μg/mL) and cell-free filtrate both showed strong therapeutic and protective effects (>70%) on grey mold both in detached tomato fruits and plants, and leaf scorch in pepper plants. This study demonstrates X. nematophila TB produces antimicrobial metabolites of strong activity on plant pathogens, with great potential for controlling tomato grey mold and pepper leaf scorch and being used in integrated disease control to reduce chemical application. PMID:24599183

  10. Beyond plant defense: insights on the potential of salicylic and methylsalicylic acid to contain growth of the phytopathogen Botrytis cinerea

    PubMed Central

    Dieryckx, Cindy; Gaudin, Vanessa; Dupuy, Jean-William; Bonneu, Marc; Girard, Vincent; Job, Dominique

    2015-01-01

    Using Botrytis cinerea we confirmed in the present work several previous studies showing that salicylic acid, a main plant hormone, inhibits fungal growth in vitro. Such an inhibitory effect was also observed for the two salicylic acid derivatives, methylsalicylic and acetylsalicylic acid. In marked contrast, 5-sulfosalicylic acid was totally inactive. Comparative proteomics from treated vs. control mycelia showed that both the intracellular and extracellular proteomes were affected in the presence of salicylic acid or methylsalicylic acid. These data suggest several mechanisms that could potentially account for the observed fungal growth inhibition, notably pH regulation, metal homeostasis, mitochondrial respiration, ROS accumulation and cell wall remodeling. The present observations support a role played by the phytohormone SA and derivatives in directly containing the pathogen. Data are available via ProteomeXchange with identifier PXD002873. PMID:26528317

  11. Pseudomonas spp.-induced systemic resistance to Botrytis cinerea is associated with induction and priming of defence responses in grapevine.

    PubMed

    Verhagen, Bas W M; Trotel-Aziz, Patricia; Couderchet, Michel; Höfte, Monica; Aziz, Aziz

    2010-01-01

    Non-pathogenic rhizobacteria Pseudomonas spp. can reduce disease in plant tissues through induction of a defence state known as induced systemic resistance (ISR). This resistance is based on multiple bacterial determinants, but nothing is known about the mechanisms underlying rhizobacteria-induced resistance in grapevine. In this study, the ability of Pseudomonas fluorescens CHA0 and Pseudomonas aeruginosa 7NSK2 to induce resistance in grapevine against Botrytis cinerea is demonstrated. Both strains also triggered an oxidative burst and phytoalexin (i.e. resveratrol and viniferin) accumulation in grape cells and primed leaves for accelerated phytoalexin production upon challenge with B. cinerea. Treatment of cell cultures with crude cell extracts of bacteria strongly enhanced oxidative burst, but resulted in comparable amounts of phytoalexins and resistance to B. cinerea to those induced by living bacteria. This suggests the production of bacterial compounds serving as inducers of disease resistance. Using other strains with different characteristics, it is shown that P. fluorescens WCS417 (Pch-deficient), P. putida WCS358 (Pch- and SA-deficient) and P. fluorescens Q2-87 (a DAPG producer) were all capable of inducing resistance to an extent similar to that induced by CHA0. However, in response to WCS417 (Pch-negative) the amount of H2O2 induced is less than for the CHA0. WCS417 induced low phytoalexin levels in cells and lost the capacity to prime for phytoalexins in the leaves. This suggests that, depending on the strain, SA, pyochelin, and DAPG are potentially effective in inducing or priming defence responses. The 7NSK2 mutants, KMPCH (Pch- and Pvd-negative) and KMPCH-567 (Pch-, Pvd-, and SA-negative) induced only partial resistance to B. cinerea. However, the amount of H2O2 triggered by KMPCH and KMPCH-567 was similar to that induced by 7NSK2. Both mutants also led to a low level of phytoalexins in grapevine cells, while KMPCH slightly primed grapevine leaves

  12. Involvement of BcStr2 in methionine biosynthesis, vegetative differentiation, multiple stress tolerance and virulence in Botrytis cinerea.

    PubMed

    Shao, Wenyong; Yang, Yalan; Zhang, Yu; Lv, Chiyuan; Ren, Weichao; Chen, Changjun

    2016-04-01

    The Str2 gene encodes a cystathionine γ-synthase that is a key enzyme in methionine (Met) biosynthesis in Saccharomyces cerevisiae. Met plays a critical role in protein synthesis and diverse cellular processes in both eukaryotes and prokaryotes. In this study, we characterized the Str2 orthologue gene BcStr2 in Botrytis cinerea. The BcStr2 mutant was unable to grow on minimal medium (MM). In addition, conidia of the mutant were unable to germinate in water-agar medium within 15 h of incubation. Supplementation with 1 mm Met or 0.5 mg/mL homocysteine, but not 1 mm cysteine or 0.5 mg/mL glutathione, rescued the defect in mycelial growth of the BcStr2 deletion mutant. These results indicate that the enzyme encoded by BcStr2 is involved in the conversion of cysteine into homocysteine. The mutant exhibited decreased conidiation and impaired sclerotium development. In addition, the BcStr2 mutant exhibited increased sensitivity to osmotic and oxidative stresses, cell wall-damaging agents and thermal stress. The mutant demonstrated dramatically decreased virulence on host plant tissues. All of the defects were restored by genetic complementation of the mutant with wild-type BcStr2. Taken together, the results of this study indicate that BcStr2 plays a critical role in the regulation of various cellular processes in B. cinerea. PMID:26176995

  13. Tomato transcriptome and mutant analyses suggest a role for plant stress hormones in the interaction between fruit and Botrytis cinerea

    PubMed Central

    Blanco-Ulate, Barbara; Vincenti, Estefania; Powell, Ann L. T.; Cantu, Dario

    2013-01-01

    Fruit–pathogen interactions are a valuable biological system to study the role of plant development in the transition from resistance to susceptibility. In general, unripe fruit are resistant to pathogen infection but become increasingly more susceptible as they ripen. During ripening, fruit undergo significant physiological and biochemical changes that are coordinated by complex regulatory and hormonal signaling networks. The interplay between multiple plant stress hormones in the interaction between plant vegetative tissues and microbial pathogens has been documented extensively, but the relevance of these hormones during infections of fruit is unclear. In this work, we analyzed a transcriptome study of tomato fruit infected with Botrytis cinerea in order to profile the expression of genes for the biosynthesis, modification and signal transduction of ethylene (ET), salicylic acid (SA), jasmonic acid (JA), and abscisic acid (ABA), hormones that may be not only involved in ripening, but also in fruit interactions with pathogens. The changes in relative expression of key genes during infection and assays of susceptibility of fruit with impaired synthesis or perception of these hormones were used to formulate hypotheses regarding the involvement of these regulators in the outcome of the tomato fruit–B. cinerea interaction. PMID:23717322

  14. Phosphoproteome profiles of the phytopathogenic fungi Alternaria brassicicola and Botrytis cinerea during exponential growth in axenic cultures.

    PubMed

    Davanture, Marlène; Dumur, Jérôme; Bataillé-Simoneau, Nelly; Campion, Claire; Valot, Benoît; Zivy, Michel; Simoneau, Philippe; Fillinger, Sabine

    2014-07-01

    This study describes the gel-free phosphoproteomic analysis of the phytopathogenic fungi Alternaria brassicicola and Botrytis cinerea grown in vitro under nonlimiting conditions. Using a combination of strong cation exchange and IMAC prior to LC-MS, we identified over 1350 phosphopeptides per fungus representing over 800 phosphoproteins. The preferred phosphorylation sites were found on serine (>80%) and threonine (>15%), whereas phosphorylated tyrosine residues were found at less than 1% in A. brassicicola and at a slightly higher ratio in B. cinerea (1.5%). Biological processes represented principally among the phoshoproteins were those involved in response and transduction of stimuli as well as in regulation of cellular and metabolic processes. Most known elements of signal transduction were found in the datasets of both fungi. This study also revealed unexpected phosphorylation sites in histidine kinases, a category overrepresented in filamentous ascomycetes compared to yeast. The data have been deposited to the ProteomeXchange database with identifier PXD000817 (http://proteomecentral.proteomexchange.org/dataset/PXD000817). PMID:24825570

  15. Suppression of the homeobox gene HDTF1 enhances resistance to Verticillium dahliae and Botrytis cinerea in cotton.

    PubMed

    Gao, Wei; Long, Lu; Xu, Li; Lindsey, Keith; Zhang, Xianlong; Zhu, Longfu

    2016-05-01

    Development of pathogen-resistant crops, such as fungus-resistant cotton, has significantly reduced chemical application and improved crop yield and quality. However, the mechanism of resistance to cotton pathogens such as Verticillium dahliae is still poorly understood. In this study, we characterized a cotton gene (HDTF1) that was isolated following transcriptome profiling during the resistance response of cotton to V. dahliae. HDTF1 putatively encodes a homeodomain transcription factor, and its expression was found to be down-regulated in cotton upon inoculation with V. dahliae and Botrytis cinerea. To characterise the involvement of HDTF1 in the response to these pathogens, we used virus-induced gene silencing (VIGS) to generate HDTF1-silenced cotton. VIGS reduction in HDTF1 expression significantly enhanced cotton plant resistance to both pathogens. HDTF1 silencing resulted in activation of jasmonic acid (JA)-mediated signaling and JA accumulation. However, the silenced plants were not altered in the accumulation of salicylic acid (SA) or the expression of marker genes associated with SA signaling. These results suggest that HDTF1 is a negative regulator of the JA pathway, and resistance to V. dahliae and B. cinerea can be engineered by activation of JA signaling. PMID:26407676

  16. The SWEET family of sugar transporters in grapevine: VvSWEET4 is involved in the interaction with Botrytis cinerea.

    PubMed

    Chong, Julie; Piron, Marie-Christine; Meyer, Sophie; Merdinoglu, Didier; Bertsch, Christophe; Mestre, Pere

    2014-12-01

    During plant development, sugar export is determinant in multiple processes such as nectar production, pollen development and long-distance sucrose transport. The plant SWEET family of sugar transporters is a recently identified protein family of sugar uniporters. In rice, SWEET transporters are the target of extracellular bacteria, which have evolved sophisticated mechanisms to modify their expression and acquire sugars to sustain their growth. Here we report the characterization of the SWEET family of sugar transporters in Vitis vinifera. We identified 17 SWEET genes in the V. vinifera 40024 genome and show that they are differentially expressed in vegetative and reproductive organs. Inoculation with the biotrophic pathogens Erysiphe necator and Plasmopara viticola did not result in significant induction of VvSWEET gene expression. However, infection with the necrotroph Botrytis cinerea triggered a strong up-regulation of VvSWEET4 expression. Further characterization of VvSWEET4 revealed that it is a glucose transporter localized in the plasma membrane that is up-regulated by inducers of reactive oxygen species and virulence factors from necrotizing pathogens. Finally, Arabidopsis knockout mutants in the orthologous AtSWEET4 were found to be less susceptible to B. cinerea. We propose that stimulation of expression of a developmentally regulated glucose uniporter by reactive oxygen species production and extensive cell death after necrotrophic fungal infection could facilitate sugar acquisition from plant cells by the pathogen. PMID:25246444

  17. Overexpression of Three Glucosinolate Biosynthesis Genes in Brassica napus Identifies Enhanced Resistance to Sclerotinia sclerotiorum and Botrytis cinerea.

    PubMed

    Zhang, Yuanyuan; Huai, Dongxin; Yang, Qingyong; Cheng, Yan; Ma, Ming; Kliebenstein, Daniel J; Zhou, Yongming

    2015-01-01

    Sclerotinia sclerotiorum and Botrytis cinerea are notorious plant pathogenic fungi with an extensive host range including Brassica crops. Glucosinolates (GSLs) are an important group of secondary metabolites characteristic of the Brassicales order, whose degradation products are proving to be increasingly important in plant protection. Enhancing the defense effect of GSL and their associated degradation products is an attractive strategy to strengthen the resistance of plants by transgenic approaches. We generated the lines of Brassica napus with three biosynthesis genes involved in GSL metabolic pathway (BnMAM1, BnCYP83A1 and BnUGT74B1), respectively. We then measured the foliar GSLs of each transgenic lines and inoculated them with S. sclerotiorum and B. cinerea. Compared with the wild type control, over-expressing BnUGT74B1 in B. napus increased the aliphatic and indolic GSL levels by 1.7 and 1.5 folds in leaves respectively; while over-expressing BnMAM1 or BnCYP83A1 resulted in an approximate 1.5-fold higher only in the aliphatic GSL level in leaves. The results of plant inoculation demonstrated that BnUGT74B1-overexpressing lines showed less severe disease symptoms and tissue damage compared with the wild type control, but BnMAM1 or BnCYP83A1-overexpressing lines showed no significant difference in comparison to the controls. These results suggest that the resistance to S. sclerotiorum and B. cinerea in B. napus could be enhanced through tailoring the GSL profiles by transgenic approaches or molecular breeding, which provides useful information to assist plant breeders to design improved breeding strategies. PMID:26465156

  18. Influence of Fungal Strain, Temperature, and Wetness Duration on Infection of Grapevine Inflorescences and Young Berry Clusters by Botrytis cinerea.

    PubMed

    Ciliberti, Nicola; Fermaud, Marc; Languasco, Luca; Rossi, Vittorio

    2015-03-01

    The effect of temperature and wetness duration on infection of Vitis vinifera inflorescences (from "inflorescence clearly visible" to "end of flowering" stages) and young berry clusters (at "fruit swelling" and "berries groat-sized" stages) by Botrytis cinerea was investigated. Artificial inoculations were carried out using conidial suspensions of eight B. cinerea strains belonging to the transposon genotypes transposa and vacuma. Infection incidence was significantly affected by strain but not by transposon genotype (transposon genotype accounted for only 6.5% of the variance). Infection incidence was also affected by the interaction between strain and growth stage of the inflorescence or berry cluster (overall accounting for approximately 57% of the experimental variance). Thus, under our experimental conditions, the ability to cause infection was a strain rather than a transposon genotype attribute. Across all strains, infection incidence was lowest when inflorescences were clearly visible or fully developed, highest at flowering (from beginning to end of flowering), and intermediate at the postflowering fruit stages (fruit swelling and berries groat-sized). One transposa strain, however, was highly virulent on all grapevine growth stages tested. The effects of temperature and wetness duration on infection incidence were similar for all fungal strains and grapevine growth stages; infection incidence was highest at 20°C and lowest at 30°C, and was also low at 5°C. Similar results were obtained for mycelial growth and conidial germination. Based on the pooled data for all strains and grapevine growth stages, an equation was developed that accounted for the combined effects of temperature and wetness duration on relative infection incidence. This equation should be useful for developing decision-making systems concerning B. cinerea control at early grapevine growth stages. PMID:25354016

  19. Overexpression of Three Glucosinolate Biosynthesis Genes in Brassica napus Identifies Enhanced Resistance to Sclerotinia sclerotiorum and Botrytis cinerea

    PubMed Central

    Yang, Qingyong; Cheng, Yan; Ma, Ming; Kliebenstein, Daniel J.; Zhou, Yongming

    2015-01-01

    Sclerotinia sclerotiorum and Botrytis cinerea are notorious plant pathogenic fungi with an extensive host range including Brassica crops. Glucosinolates (GSLs) are an important group of secondary metabolites characteristic of the Brassicales order, whose degradation products are proving to be increasingly important in plant protection. Enhancing the defense effect of GSL and their associated degradation products is an attractive strategy to strengthen the resistance of plants by transgenic approaches. We generated the lines of Brassica napus with three biosynthesis genes involved in GSL metabolic pathway (BnMAM1, BnCYP83A1 and BnUGT74B1), respectively. We then measured the foliar GSLs of each transgenic lines and inoculated them with S. sclerotiorum and B. cinerea. Compared with the wild type control, over-expressing BnUGT74B1 in B. napus increased the aliphatic and indolic GSL levels by 1.7 and 1.5 folds in leaves respectively; while over-expressing BnMAM1 or BnCYP83A1 resulted in an approximate 1.5-fold higher only in the aliphatic GSL level in leaves. The results of plant inoculation demonstrated that BnUGT74B1-overexpressing lines showed less severe disease symptoms and tissue damage compared with the wild type control, but BnMAM1 or BnCYP83A1-overexpressing lines showed no significant difference in comparison to the controls. These results suggest that the resistance to S. sclerotiorum and B. cinerea in B. napus could be enhanced through tailoring the GSL profiles by transgenic approaches or molecular breeding, which provides useful information to assist plant breeders to design improved breeding strategies. PMID:26465156

  20. Dark Period Following UV-C Treatment Enhances Killing of Botrytis cinerea Conidia and Controls Gray Mold of Strawberries.

    PubMed

    Janisiewicz, Wojciech J; Takeda, Fumiomi; Glenn, D Michael; Camp, Mary J; Jurick, Wayne M

    2016-04-01

    Strawberries are available throughout the year either from production in the field or from high and low tunnel culture. Diversity of production conditions results in new challenges in controlling diseases before and after harvest. Fungicides have traditionally been used to control these diseases; however, their limitations necessitate a search for new approaches. We found that UV-C irradiation of Botrytis cinerea, a major pathogen of strawberry, can effectively kill this fungus if a dark period follows the treatment. The inclusion of a 4-h dark period resulted in almost complete kill of B. cinerea conidia on agar media at a dose of 12.36 J/m2. The UV-C dose did not cause a reduction in photosynthesis in strawberry leaves or discoloration of sepals, even after exposing plants repeatedly (twice a week) for 7 weeks. Although irradiation of dry conidia of B. cinerea with this dose resulted in some survival, the conidia were not infective and not able to cause decay even when inoculated onto a highly susceptible mature apple fruit. Irradiation of strawberry pollen at 12.36 J/m2 did not affect pollen germination, tube growth and length in vitro, or germination and tube growth in the style of hand-pollinated emasculated strawberry flowers. No negative effect of the UV-C treatment was observed on fruit yield and quality in high tunnel culture. In the fruit and flower petal inoculation tests, the UV-C treatment was highly effective in reducing fruit decay and petal infection. This UV-C treatment with an exposure time of 60 s may be useful in controlling gray mold in tunnel production of strawberries and may also have the potential for use in intensive field and indoor production of other fruits and vegetables providing that a 4-h dark period follows the irradiation. PMID:26714103

  1. Synergistic effect of the combined treatment with gamma irradiation and sodium dichloroisocyanurate to control gray mold (Botrytis cinerea) on paprika

    NASA Astrophysics Data System (ADS)

    Yoon, Minchul; Jung, Koo; Lee, Kwang-Youll; Jeong, Je-Yong; Lee, Ju-Woon; Park, Hae-Jun

    2014-05-01

    Gray mold (Botrytis cinerea) is one of the most major fungal pathogens in paprika. Generally, gamma irradiation over 1 kGy is effective for the control of fungal pathogens; however, a significant change in fruit quality (physical properties) on paprika was shown from gamma irradiation at over 0.6 kGy (p<0.05). Therefore, in this study, the synergistic disinfection effect of the combined treatment with gamma irradiation and sodium dichloroisocyanurate (NaDCC) was investigated to reduce the gamma irradiation dose. In an artificial inoculation experiment of B. cinerea isolated from naturally-infected postharvest paprika, fungal symptoms were observed in the stem and exocarp of paprika after conidial inoculation. From the sensitivity of gamma irradiation and NaDCC, B. cinerea conidia were fully inactivated by 4 kGy of gamma irradiation (D10 value 0.99 kGy), and were fully inactivated by 50 ppm NaDCC treatment. The fungal symptoms were not detected by the dose-dependent gamma irradiation (>4 kGy) and NaDCC (>50 ppm). As a result of the combined treatment of gamma irradiation and NaDCC, the D10 value was significantly reduced by 1.06, 0.88, 0.77, and 0.58 kGy (p<0.05). Moreover, fungal symptoms were more significantly reduced in combined treatment groups (gamma irradiation and NaDCC) than single treatment groups (gamma irradiation or NaDCC). These results suggest that combined treatment with irradiation and NaDCC treatment can be applied to preserve quality of postharvest paprika or other fruits.

  2. Tomato SlRbohB, a member of the NADPH oxidase family, is required for disease resistance against Botrytis cinerea and tolerance to drought stress.

    PubMed

    Li, Xiaohui; Zhang, Huijuan; Tian, Limei; Huang, Lei; Liu, Shixia; Li, Dayong; Song, Fengming

    2015-01-01

    NADPH oxidases (also known as respiratory burst oxidase homologs, Rbohs) are key enzymes that catalyze the generation of reactive oxygen species (ROS) in plants. In the present study, eight SlRboh genes were identified in tomato and their possible involvement in resistance to Botrytis cinerea and drought tolerance was examined. Expression of SlRbohs was induced by B. cinerea and Pseudomonas syringae pv. tomato but displayed distinct patterns. Virus-induced gene silencing based silencing of SlRbohB resulted in reduced resistance to B. cinerea but silencing of other SlRbohs did not affect the resistance. Compared to non-silenced plants, the SlRbohB-silenced plants accumulated more ROS and displayed attenuated expression of defense genes after infection with B. cinerea. Silencing of SlRbohB also suppressed flg22-induced ROS burst and the expression of SlLrr22, a marker gene related to PAMP-triggered immunity (PTI). Transient expression of SlRbohB in Nicotiana benthamiana led to enhanced resistance to B. cinerea. Furthermore, silencing of SlRbohB resulted in decreased drought tolerance, accelerated water loss in leaves and the altered expression of drought-responsive genes. Our data demonstrate that SlRbohB positively regulates the resistance to B. cinerea, flg22-induced PTI, and drought tolerance in tomato. PMID:26157450

  3. Living Colors in the Gray Mold Pathogen Botrytis cinerea: Codon-Optimized Genes Encoding Green Fluorescent Protein and mCherry, Which Exhibit Bright Fluorescence▿

    PubMed Central

    Leroch, Michaela; Mernke, Dennis; Koppenhoefer, Dieter; Schneider, Prisca; Mosbach, Andreas; Doehlemann, Gunther; Hahn, Matthias

    2011-01-01

    The green fluorescent protein (GFP) and its variants have been widely used in modern biology as reporters that allow a variety of live-cell imaging techniques. So far, GFP has rarely been used in the gray mold fungus Botrytis cinerea because of low fluorescence intensity. The codon usage of B. cinerea genes strongly deviates from that of commonly used GFP-encoding genes and reveals a lower GC content than other fungi. In this study, we report the development and use of a codon-optimized version of the B. cinerea enhanced GFP (eGFP)-encoding gene (Bcgfp) for improved expression in B. cinerea. Both the codon optimization and, to a smaller extent, the insertion of an intron resulted in higher mRNA levels and increased fluorescence. Bcgfp was used for localization of nuclei in germinating spores and for visualizing host penetration. We further demonstrate the use of promoter-Bcgfp fusions for quantitative evaluation of various toxic compounds as inducers of the atrB gene encoding an ABC-type drug efflux transporter of B. cinerea. In addition, a codon-optimized mCherry-encoding gene was constructed which yielded bright red fluorescence in B. cinerea. PMID:21378036

  4. Gray Mold Populations in German Strawberry Fields Are Resistant to Multiple Fungicides and Dominated by a Novel Clade Closely Related to Botrytis cinerea

    PubMed Central

    Leroch, Michaela; Plesken, Cecilia; Weber, Roland W. S.; Kauff, Frank; Scalliet, Gabriel

    2013-01-01

    The gray mold fungus Botrytis cinerea is a major threat to fruit and vegetable production. Strawberry fields usually receive several fungicide treatments against Botrytis per season. Gray mold isolates from several German strawberry-growing regions were analyzed to determine their sensitivity against botryticides. Fungicide resistance was commonly observed, with many isolates possessing resistance to multiple (up to six) fungicides. A stronger variant of the previously described multidrug resistance (MDR) phenotype MDR1, called MDR1h, was found to be widely distributed, conferring increased partial resistance to two important botryticides, cyprodinil and fludioxonil. A 3-bp deletion mutation in a transcription factor-encoding gene, mrr1, was found to be correlated with MDR1h. All MDR1h isolates and the majority of isolates with resistance to multiple fungicides were found to be genetically distinct. Multiple-gene sequencing confirmed that they belong to a novel clade, called Botrytis group S, which is closely related to B. cinerea and the host-specific species B. fabae. Isolates of Botrytis group S genotypes were found to be widespread in all German strawberry-growing regions but almost absent from vineyards. Our data indicate a clear subdivision of gray mold populations, which are differentially distributed according to their host preference and adaptation to chemical treatments. PMID:23087030

  5. Construction of a Streptomyces lydicus A01 transformant with a chit42 gene from Trichoderma harzianum P1 and evaluation of its biocontrol activity against Botrytis cinerea.

    PubMed

    Wu, Qiong; Bai, Linquan; Liu, Weicheng; Li, Yingying; Lu, Caige; Li, Yaqian; Fu, Kehe; Yu, Chuanjin; Chen, Jie

    2013-04-01

    Streptomyces lydicus A01 and Trichoderma harzianum P1 are potential biocontrol agents of fungal diseases in plants. S. lydicus A01 produces natamycin to bind the ergosterol of the fungal cell membrane and inhibits the growth of Botrytis cinerea. T. harzianum P1, on the other hand, features high chitinase activity and decomposes the chitin in the cell wall of B. cinerea. To obtain the synergistic biocontrol effects of chitinase and natamycin on Botrytis cinerea, this study transformed the chit42 gene from T. harzianum P1 to S. lydicus A01. The conjugal transformant (CT) of S. lydicus A01 with the chit42 gene was detected using polymerase chain reaction (PCR). Associated chitinase activity and natamycin production were examined using the 3, 5-dinitrosalicylic acid (DNS) method and ultraviolet spectrophotometry, respectively. The S. lydicus A01-chit42 CT showed substantially higher chitinase activity and natamycin production than its wild type strain (WT). Consequently, the biocontrol effects of S. lydicus A01-chit42 CT on B. cinerea, including inhibition to spore germination and mycelial growth, were highly improved compared with those of the WT. Our research indicates that the biocontrol effect of Streptomyces can be highly improved by transforming the exogenous resistance gene, i.e. chit42 from Trichoderma, which not only enhances the production of antibiotics, but also provides a supplementary function by degrading the cell walls of the pathogens. PMID:23625216

  6. Oviposition preference and larval performance of Epiphyas postvittana (Lepidoptera: Tortricidae) on Botrytis cinerea (Helotiales: Sclerotiniaceae) infected berries of Vitis vinifera (Vitales: Vitaceae).

    PubMed

    Rizvi, Syed Z M; Raman, Anantanarayanan; Wheatley, Warwick M; Cook, Geoffrey

    2016-04-01

    In this paper we tested the behavior of gravid Epiphyas postvittana in selecting the most-appropriate site for oviposition thus benefitting offspring performance. Our hypothesis was built on Jaenike's preference-performance hypothesis (also referred to as the "mother-knows-the-best" hypothesis). To test this, we used the interacting Epiphyas postvittana, its host Vitis vinifera, and the pathogenic microbe Botrytis cinerea system. Populations of E. postvittana and B. cinerea often exist concurrently on V. vinifera in Australasia and their interaction and mutual influence are currently being explored, although the suggestion presently is that the relationship between E. postvittana and B. cinerea is mutualistic. We tested the effect of volatiles from B. cinerea-infected berries and uninfected (control) berries of V. vinifera on the oviposition behavior of E. postvittana. We also characterized the effects of B. cinerea infection on the berries of V. vinifera on the growth and development of E. postvittana. Contrary to the preference-performance hypothesis, oviposition choices made by gravid E. postvittana did not result in the best offspring survival, development, and performance. The preference for oviposition by E. postvittana was strongly influenced by the olfactory and tactile cues. She laid fewer eggs on B. cinerea-infected berries compared to uninfected berries of V. vinifera. The larvae of E. postvittana showed no preference to uninfected berries of V. vinifera. The larvae fed on B. cinerea-infected berries of V. vinifera showing greater survival rate, shorter time to pupation, greater pupal mass, and on becoming adults they laid more numbers of eggs than the larvae that were enabled to feed on uninfected berries. The larvae of E. postvittana transport the conidia of B. cinerea and transmit grey-mould disease to uninfected berries of V. vinifera. PMID:25420720

  7. Resistance evaluation of Chinese wild Vitis genotypes against Botrytis cinerea and different responses of resistant and susceptible hosts to the infection

    PubMed Central

    Wan, Ran; Hou, Xiaoqing; Wang, Xianhang; Qu, Jingwu; Singer, Stacy D.; Wang, Yuejin; Wang, Xiping

    2015-01-01

    The necrotrophic fungus Botrytis cinerea is a major threat to grapevine cultivation worldwide. A screen of 41 Vitis genotypes for leaf resistance to B. cinerea suggested species independent variation and revealed 18 resistant Chinese wild Vitis genotypes, while most investigated V. vinifera, or its hybrids, were susceptible. A particularly resistant Chinese wild Vitis, “Pingli-5” (V. sp. [Qinling grape]) and a very susceptible V. vinifera cultivar, “Red Globe” were selected for further study. Microscopic analysis demonstrated that B. cinerea growth was limited during early infection on “Pingli-5” before 24 h post-inoculation (hpi) but not on Red Globe. It was found that reactive oxygen species (ROS) and antioxidative system were associated with fungal growth. O2- accumulated similarly in B. cinerea 4 hpi on both Vitis genotypes. Lower levels of O2- (not H2O2) were detected 4 hpi and ROS (H2O2 and O2-) accumulation from 8 hpi onwards was also lower in “Pingli-5” leaves than in “Red Globe” leaves. B. cinerea triggered sustained ROS production in “Red Globe” but not in “Pingli-5” with subsequent infection progresses. Red Globe displayed little change in antioxidative activities in response to B. cinerea infection, instead, antioxidative activities were highly and timely elevated in resistant “Pingli-5” which correlated with its minimal ROS increases and its high resistance. These findings not only enhance our understanding of the resistance of Chinese wild Vitis species to B. cinerea, but also lay the foundation for breeding B. cinerea resistant grapes in the future. PMID:26579134

  8. Environmental Conditions Affect Botrytis cinerea Infection of Mature Grape Berries More Than the Strain or Transposon Genotype.

    PubMed

    Ciliberti, Nicola; Fermaud, Marc; Roudet, Jean; Rossi, Vittorio

    2015-08-01

    Effects of environment, Botrytis cinerea strain, and their interaction on the infection of mature grape berries were investigated. The combined effect of temperature (T) of 15, 20, 25, and 30°C and relative humidity (RH) of 65, 80, 90, and 100% was studied by inoculating berries with mycelium plugs. Regardless of the T, no disease occurred at 65% RH, and both disease incidence and severity increased with increasing RH. The combined effect of T (5 to 30°C) and wetness duration (WD) of 3, 6, 12, 24, and 36 h was studied by inoculating berries with conidia. At WD of 36 h, disease incidence was approximately 75% of affected berries at 20 or 25°C, 50% at 15°C, and 30 to 20% at 30 and 10°C; no infection occurred at 5°C. Under favorable conditions (100% RH or 36 h of WD) and unfavorable conditions (65% RH or 3 h of WD), berry wounding did not significantly affect disease incidence; under moderately favorable conditions (80% RH or 6 to 12 h of WD), disease incidence was approximately 1.5 to 5 times higher in wounded than in intact berries. Our data collectively showed that (i) T and RH or WD were more important than strain for mature berry infection by either mycelium or conidia and (ii) the effect of the environment on the different strains was similar. Two equations were developed describing the combined effect of T and RH, or T and WD, on disease incidence following inoculation by mycelium (R2=0.99) or conidia (R2=0.96), respectively. These equations may be useful in the development of models used to predict and control Botrytis bunch rot during berry ripening. PMID:26218433

  9. Developmental and Metabolic Plasticity of White-Skinned Grape Berries in Response to Botrytis cinerea during Noble Rot1[OPEN

    PubMed Central

    Collins, Thomas S.; Vicente, Ariel R.; Doyle, Carolyn L.; Ye, Zirou; Allen, Greg; Heymann, Hildegarde

    2015-01-01

    Noble rot results from exceptional infections of ripe grape (Vitis vinifera) berries by Botrytis cinerea. Unlike bunch rot, noble rot promotes favorable changes in grape berries and the accumulation of secondary metabolites that enhance wine grape composition. Noble rot-infected berries of cv Sémillon, a white-skinned variety, were collected over 3 years from a commercial vineyard at the same time that fruit were harvested for botrytized wine production. Using an integrated transcriptomics and metabolomics approach, we demonstrate that noble rot alters the metabolism of cv Sémillon berries by inducing biotic and abiotic stress responses as well as ripening processes. During noble rot, B. cinerea induced the expression of key regulators of ripening-associated pathways, some of which are distinctive to the normal ripening of red-skinned cultivars. Enhancement of phenylpropanoid metabolism, characterized by a restricted flux in white-skinned berries, was a common outcome of noble rot and red-skinned berry ripening. Transcript and metabolite analyses together with enzymatic assays determined that the biosynthesis of anthocyanins is a consistent hallmark of noble rot in cv Sémillon berries. The biosynthesis of terpenes and fatty acid aroma precursors also increased during noble rot. We finally characterized the impact of noble rot in botrytized wines. Altogether, the results of this work demonstrated that noble rot causes a major reprogramming of berry development and metabolism. This desirable interaction between a fruit and a fungus stimulates pathways otherwise inactive in white-skinned berries, leading to a greater accumulation of compounds involved in the unique flavor and aroma of botrytized wines. PMID:26450706

  10. Natural Variation in the VELVET Gene bcvel1 Affects Virulence and Light-Dependent Differentiation in Botrytis cinerea

    PubMed Central

    Schumacher, Julia; Pradier, Jean-Marc; Simon, Adeline; Traeger, Stefanie; Moraga, Javier; Collado, Isidro González; Viaud, Muriel; Tudzynski, Bettina

    2012-01-01

    Botrytis cinerea is an aggressive plant pathogen causing gray mold disease on various plant species. In this study, we identified the genetic origin for significantly differing phenotypes of the two sequenced B. cinerea isolates, B05.10 and T4, with regard to light-dependent differentiation, oxalic acid (OA) formation and virulence. By conducting a map-based cloning approach we identified a single nucleotide polymorphism (SNP) in an open reading frame encoding a VELVET gene (bcvel1). The SNP in isolate T4 results in a truncated protein that is predominantly found in the cytosol in contrast to the full-length protein of isolate B05.10 that accumulates in the nuclei. Deletion of the full-length gene in B05.10 resulted in the T4 phenotype, namely light-independent conidiation, loss of sclerotial development and oxalic acid production, and reduced virulence on several host plants. These findings indicate that the identified SNP represents a loss-of-function mutation of bcvel1. In accordance, the expression of the B05.10 copy in T4 rescued the wild-type/B05.10 phenotype. BcVEL1 is crucial for full virulence as deletion mutants are significantly hampered in killing and decomposing plant tissues. However, the production of the two best known secondary metabolites, the phytotoxins botcinic acid and botrydial, are not affected by the deletion of bcvel1 indicating that other factors are responsible for reduced virulence. Genome-wide expression analyses of B05.10- and Δbcvel1-infected plant material revealed a number of genes differentially expressed in the mutant: while several protease- encoding genes are under-expressed in Δbcvel1 compared to the wild type, the group of over-expressed genes is enriched for genes encoding sugar, amino acid and ammonium transporters and glycoside hydrolases reflecting the response of Δbcvel1 mutants to nutrient starvation conditions. PMID:23118899

  11. A functional bikaverin biosynthesis gene cluster in rare strains of Botrytis cinerea is positively controlled by VELVET.

    PubMed

    Schumacher, Julia; Gautier, Angélique; Morgant, Guillaume; Studt, Lena; Ducrot, Paul-Henri; Le Pêcheur, Pascal; Azeddine, Saad; Fillinger, Sabine; Leroux, Pierre; Tudzynski, Bettina; Viaud, Muriel

    2013-01-01

    The gene cluster responsible for the biosynthesis of the red polyketidic pigment bikaverin has only been characterized in Fusarium ssp. so far. Recently, a highly homologous but incomplete and nonfunctional bikaverin cluster has been found in the genome of the unrelated phytopathogenic fungus Botrytis cinerea. In this study, we provided evidence that rare B. cinerea strains such as 1750 have a complete and functional cluster comprising the six genes orthologous to Fusarium fujikuroi ffbik1-ffbik6 and do produce bikaverin. Phylogenetic analysis confirmed that the whole cluster was acquired from Fusarium through a horizontal gene transfer (HGT). In the bikaverin-nonproducing strain B05.10, the genes encoding bikaverin biosynthesis enzymes are nonfunctional due to deleterious mutations (bcbik2-3) or missing (bcbik1) but interestingly, the genes encoding the regulatory proteins BcBIK4 and BcBIK5 do not harbor deleterious mutations which suggests that they may still be functional. Heterologous complementation of the F. fujikuroi Δffbik4 mutant confirmed that bcbik4 of strain B05.10 is indeed fully functional. Deletion of bcvel1 in the pink strain 1750 resulted in loss of bikaverin and overproduction of melanin indicating that the VELVET protein BcVEL1 regulates the biosynthesis of the two pigments in an opposite manner. Although strain 1750 itself expresses a truncated BcVEL1 protein (100 instead of 575 aa) that is nonfunctional with regard to sclerotia formation, virulence and oxalic acid formation, it is sufficient to regulate pigment biosynthesis (bikaverin and melanin) and fenhexamid HydR2 type of resistance. Finally, a genetic cross between strain 1750 and a bikaverin-nonproducing strain sensitive to fenhexamid revealed that the functional bikaverin cluster is genetically linked to the HydR2 locus. PMID:23308280

  12. Fungicide-Driven Evolution and Molecular Basis of Multidrug Resistance in Field Populations of the Grey Mould Fungus Botrytis cinerea

    PubMed Central

    Mosbach, Andreas; Walker, Anne-Sophie; Fillinger, Sabine; Mernke, Dennis; Schoonbeek, Henk-Jan; Pradier, Jean-Marc; Leroux, Pierre; De Waard, Maarten A.; Hahn, Matthias

    2009-01-01

    The grey mould fungus Botrytis cinerea causes losses of commercially important fruits, vegetables and ornamentals worldwide. Fungicide treatments are effective for disease control, but bear the risk of resistance development. The major resistance mechanism in fungi is target protein modification resulting in reduced drug binding. Multiple drug resistance (MDR) caused by increased efflux activity is common in human pathogenic microbes, but rarely described for plant pathogens. Annual monitoring for fungicide resistance in field isolates from fungicide-treated vineyards in France and Germany revealed a rapidly increasing appearance of B. cinerea field populations with three distinct MDR phenotypes. All MDR strains showed increased fungicide efflux activity and overexpression of efflux transporter genes. Similar to clinical MDR isolates of Candida yeasts that are due to transcription factor mutations, all MDR1 strains were shown to harbor activating mutations in a transcription factor (Mrr1) that controls the gene encoding ABC transporter AtrB. MDR2 strains had undergone a unique rearrangement in the promoter region of the major facilitator superfamily transporter gene mfsM2, induced by insertion of a retrotransposon-derived sequence. MDR2 strains carrying the same rearranged mfsM2 allele have probably migrated from French to German wine-growing regions. The roles of atrB, mrr1 and mfsM2 were proven by the phenotypes of knock-out and overexpression mutants. As confirmed by sexual crosses, combinations of mrr1 and mfsM2 mutations lead to MDR3 strains with higher broad-spectrum resistance. An MDR3 strain was shown in field experiments to be selected against sensitive strains by fungicide treatments. Our data document for the first time the rising prevalence, spread and molecular basis of MDR populations in a major plant pathogen in agricultural environments. These populations will increase the risk of grey mould rot and hamper the effectiveness of current strategies for

  13. A Functional Bikaverin Biosynthesis Gene Cluster in Rare Strains of Botrytis cinerea Is Positively Controlled by VELVET

    PubMed Central

    Schumacher, Julia; Gautier, Angélique; Morgant, Guillaume; Studt, Lena; Ducrot, Paul-Henri; Le Pêcheur, Pascal; Azeddine, Saad; Fillinger, Sabine; Leroux, Pierre; Tudzynski, Bettina; Viaud, Muriel

    2013-01-01

    The gene cluster responsible for the biosynthesis of the red polyketidic pigment bikaverin has only been characterized in Fusarium ssp. so far. Recently, a highly homologous but incomplete and nonfunctional bikaverin cluster has been found in the genome of the unrelated phytopathogenic fungus Botrytis cinerea. In this study, we provided evidence that rare B. cinerea strains such as 1750 have a complete and functional cluster comprising the six genes orthologous to Fusarium fujikuroi ffbik1-ffbik6 and do produce bikaverin. Phylogenetic analysis confirmed that the whole cluster was acquired from Fusarium through a horizontal gene transfer (HGT). In the bikaverin-nonproducing strain B05.10, the genes encoding bikaverin biosynthesis enzymes are nonfunctional due to deleterious mutations (bcbik2-3) or missing (bcbik1) but interestingly, the genes encoding the regulatory proteins BcBIK4 and BcBIK5 do not harbor deleterious mutations which suggests that they may still be functional. Heterologous complementation of the F. fujikuroi Δffbik4 mutant confirmed that bcbik4 of strain B05.10 is indeed fully functional. Deletion of bcvel1 in the pink strain 1750 resulted in loss of bikaverin and overproduction of melanin indicating that the VELVET protein BcVEL1 regulates the biosynthesis of the two pigments in an opposite manner. Although strain 1750 itself expresses a truncated BcVEL1 protein (100 instead of 575 aa) that is nonfunctional with regard to sclerotia formation, virulence and oxalic acid formation, it is sufficient to regulate pigment biosynthesis (bikaverin and melanin) and fenhexamid HydR2 type of resistance. Finally, a genetic cross between strain 1750 and a bikaverin-nonproducing strain sensitive to fenhexamid revealed that the functional bikaverin cluster is genetically linked to the HydR2 locus. PMID:23308280

  14. Chemically Induced Cryptic Sesquiterpenoids and Expression of Sesquiterpene Cyclases in Botrytis cinerea Revealed New Sporogenic (+)-4-Epieremophil-9-en-11-ols.

    PubMed

    Pinedo, Cristina; Moraga, Javier; Barua, Javier; González-Rodríguez, Victoria E; Aleu, Josefina; Durán-Patrón, Rosa; Macías-Sánchez, Antonio J; Hanson, James R; Viaud, Muriel; Hernández-Galán, Rosario; Garrido, Carlos; Collado, Isidro G

    2016-05-20

    The sequencing of the genomes of the B05.10 and T4 strains of the fungus Botrytis cinerea revealed an abundance of novel biosynthetic gene clusters, the majority of which were unexpected on the basis of the previous analyses of the fermentation of these and closely related species. By systematic alteration of easy accessible cultivation parameters, using chemical induction with copper sulfate, we have found a cryptic sesquiterpenoid family with new structures related to eremophil-9-ene, which had the basic structure of the sesquiterpene (+)-5-epiaristolochene ((+)-4-epieremophil-9-ene). An expression study of the sesquiterpene cyclase genes present in the Botrytis cinerea genome, under culture conditions, is reported. In general, a 3 day delay and a higher BcSTC genes expression were observed when copper (5 ppm) was fed to the fermentation broth. In addition, to the observed effect on the BcBOT2 (BcSTC1) gene, involved in the biosynthesis of the botrydial toxin, a higher expression level for BcSTC3 and BcSTC4 was observed with respect to the control in the strain B05.10. Interestingly, under copper conditions, the BcSTC4 gene was the most expressed gene in the Botrytis cinerea UCA992 strain. In vitro evaluation of the biological role of these metabolites indicates that they contributed to the conidial development in B. cinerea and appear to be involved in self-regulation of the production of asexual spores. Furthermore, they promoted the formation of complex appressoria or infection cushions. PMID:26900713

  15. Pseudomonas fluorescens PTA-CT2 Triggers Local and Systemic Immune Response Against Botrytis cinerea in Grapevine.

    PubMed

    Gruau, Charlotte; Trotel-Aziz, Patricia; Villaume, Sandra; Rabenoelina, Fanja; Clément, Christophe; Baillieul, Fabienne; Aziz, Aziz

    2015-10-01

    Although induced systemic resistance (ISR) is well-documented in the context of plant-beneficial bacteria interactions, knowledge about the local and systemic molecular and biochemical defense responses before or upon pathogen infection in grapevine is very scarce. In this study, we first investigated the capacity of grapevine plants to express immune responses at both above- and below-ground levels upon interaction with a beneficial bacterium, Pseudomonas fluorescens PTA-CT2. We then explored whether the extent of priming state could contribute to the PTA-CT2-induced ISR in Botrytis cinerea-infected leaves. Our data provide evidence that this bacterium colonized grapevine roots but not the above-ground plant parts and altered the plant phenotype that displayed multiple defense responses both locally and systemically. The grapevine roots and leaves exhibited distinct patterns of defense-related gene expression during root colonization by PTA-CT2. Roots responded faster than leaves and some responses were more strongly upregulated in roots than in leaves and vice versa for other genes. These responses appear to be associated with some induction of cell death in roots and a transient expression of HSR, a hypersensitive response-related gene in both local (roots) and systemic (leaves) tissues. However, stilbenic phytoalexin patterns followed opposite trends in roots compared with leaves but no phytoalexin was exuded during plant-bacterium interaction, suggesting that roots could play an important role in the transfer of metabolites contributing to immune response at the systemic level. Unexpectedly, in B. cinerea-infected leaves PTA-CT2-mediated ISR was accompanied in large part by a downregulation of different defense-related genes, including HSR. Only phytoalexins and glutathion-S-transferase 1 transcripts were upregulated, while the expression of anthocyanin biosynthetic genes was maintained at a higher level than the control. This suggests that decreased

  16. Bis(2,3-dibromo-4,5-dihydroxybenzyl) ether, a marine algae derived bromophenol, inhibits the growth of Botrytis cinerea and interacts with DNA molecules.

    PubMed

    Liu, Ming; Wang, Genzhu; Xiao, Lin; Xu, Xuanli; Liu, Xiaohui; Xu, Pingxiang; Lin, Xiukun

    2014-07-01

    Bis(2,3-dibromo-4,5-dihydroxybenzyl) ether (BDDE) is a bromophenol isolated from marine algae. Previous reports have shown that BDDE possesses cytotoxic and antibacterial activity. In the present study, we demonstrate that BDDE displays broad-spectrum antifungal activities, especially on Botrytis cinerea. BDDE inhibits the growth of B. cinerea cultured on a solid medium of potato dextrose agar (PDA) as well as on the potato dextrose broth (PDB) medium. Moreover, BDDE decreases the incidence of fruit decay and severity of strawberries infected with B. cinerea. Further studies have revealed that BDDE decreases the germination rate and inhibits the mycelial growth of B. cinerea. The inhibition mechanisms are related to the disruption of the cell membrane integrity in B. cinerea spores and newly formed germ tubes. This study also suggests that BDDE possibly interacts with DNA via intercalation and minor groove binding. The studies provide evidence that BDDE has potential application in the control of gray mold after fruit harvest and the compound could serve as a candidate or lead template for rational drug design and for the development of antifungal agents. PMID:24979270

  17. A wild-type Botrytis cinerea strain co-infected by double-stranded RNA mycoviruses presents hypovirulence-associated traits

    PubMed Central

    2013-01-01

    Background Botrytis cinerea CCg378 is a wild-type strain infected with two types of double-stranded RNA (dsRNA) mycoviruses and which presents hypovirulence-associated traits. The objectives of the present study were to characterize the mycoviruses and investigate their relationship with the low virulence degree of the fungal host. Results B. cinerea CCg378 contains five dsRNA molecules that are associated with two different types of isometric viral particles of 32 and 23 nm in diameter, formed by structural polypeptides of 70-kDa and 48-kDa, respectively. The transfection of spheroplasts of a virus-free strain, B. cinerea CKg54, with viral particles purified from the CCg378 strain revealed that the 2.2-kbp dsRNAs have no dependency on the smaller molecules for its stable maintenance in the fungal cytoplasm, because a fungal clone that only contains the 2.2-kbp dsRNAs associated with the 32-nm particles was obtained, which we named B. cinerea CKg54vi378. One of the 2.2 kbpdsRNA segments (2219 bp) was sequenced and corresponds to the gene encoding the capsid protein of B. cinerea CCg378 virus 1 (Bc378V1), a putative new member of the Partitiviridae family. Furthermore, physiological parameters related to the degree of virulence of the fungus, such as the sporulation rate and laccase activity, were lower in B. cinerea CCg378 and B. cinerea CKg54vi378 than in B. cinerea CKg54. Additionally, bioassays performed on grapevine leaves showed that the CCg378 and CKg54vi378 strains presented a lower degree of invasiveness on the plant tissue than the CKg54 strain. Conclusions The results show that B. cinerea CCg378 is coinfected by two mycoviruses and that the 2.2-kbp dsRNAs correspond to the 32-nm mycovirus genome, which would be a new member of the Partitiviridae family as it has the typical pattern of partitiviruses. On the other hand, the results suggest that the hypovirulence of B. cinerea CCg378 could be conferred by both mycoviruses, since the fungal clone B

  18. Fast detection of peroxidase (POD) activity in tomato leaves which infected with Botrytis cinerea using hyperspectral imaging

    NASA Astrophysics Data System (ADS)

    Kong, Wenwen; Liu, Fei; Zhang, Chu; Bao, Yidan; Yu, Jiajia; He, Yong

    2014-01-01

    Tomatoes are cultivated around the world and gray mold is one of its most prominent and destructive diseases. An early disease detection method can decrease losses caused by plant diseases and prevent the spread of diseases. The activity of peroxidase (POD) is very important indicator of disease stress for plants. The objective of this study is to examine the possibility of fast detection of POD activity in tomato leaves which infected with Botrytis cinerea using hyperspectral imaging data. Five pre-treatment methods were investigated. Genetic algorithm-partial least squares (GA-PLS) was applied to select optimal wavelengths. A new fast learning neural algorithm named extreme learning machine (ELM) was employed as multivariate analytical tool in this study. 21 optimal wavelengths were selected by GA-PLS and used as inputs of three calibration models. The optimal prediction result was achieved by ELM model with selected wavelengths, and the r and RMSEP in validation were 0.8647 and 465.9880 respectively. The results indicated that hyperspectral imaging could be considered as a valuable tool for POD activity prediction. The selected wavelengths could be potential resources for instrument development.

  19. Population structure and temporal maintenance of the multihost fungal pathogen Botrytis cinerea: causes and implications for disease management.

    PubMed

    Walker, Anne-Sophie; Gladieux, Pierre; Decognet, Véronique; Fermaud, Marc; Confais, Johann; Roudet, Jean; Bardin, Marc; Bout, Alexandre; Nicot, Philippe C; Poncet, Christine; Fournier, Elisabeth

    2015-04-01

    Understanding the causes of population subdivision is of fundamental importance, as studying barriers to gene flow between populations may reveal key aspects of the process of adaptive divergence and, for pathogens, may help forecasting disease emergence and implementing sound management strategies. Here, we investigated population subdivision in the multihost fungus Botrytis cinerea based on comprehensive multiyear sampling on different hosts in three French regions. Analyses revealed a weak association between population structure and geography, but a clear differentiation according to the host plant of origin. This was consistent with adaptation to hosts, but the distribution of inferred genetic clusters and the frequency of admixed individuals indicated a lack of strict host specificity. Differentiation between individuals collected in the greenhouse (on Solanum) and outdoor (on Vitis and Rubus) was stronger than that observed between individuals from the two outdoor hosts, probably reflecting an additional isolating effect associated with the cropping system. Three genetic clusters coexisted on Vitis but did not persist over time. Linkage disequilibrium analysis indicated that outdoor populations were regularly recombining, whereas clonality was predominant in the greenhouse. Our findings open up new perspectives for disease control by managing plant debris in outdoor conditions and reinforcing prophylactic measures indoor. PMID:25040694

  20. BcNoxD, a putative ER protein, is a new component of the NADPH oxidase complex in Botrytis cinerea.

    PubMed

    Siegmund, Ulrike; Marschall, Robert; Tudzynski, Paul

    2015-03-01

    NADPH oxidases (Nox) are major enzymatic producer of reactive oxygen species (ROS). In fungi these multi-enzyme complexes are involved in sexual differentiation and pathogenicity. However, in contrast to mammalian systems, the composition and recruitment of the fungal Nox complexes are unresolved. Here we introduce a new Nox component, the membrane protein NoxD in the grey mold fungus Botrytis cinerea. It has high homology to the ER protein Pro41 from Sordaria macrospora, similar functions to the catalytic Nox subunit BcNoxA in differentiation and pathogenicity, and shows similarities to phagocytic p22phox. BcNoxA and BcNoxD interact with each other. Both proteins are involved in pathogenicity, fusion of conidial anastomosis tubes (CAT) and formation of sclerotia and conidia. These data support our earlier view based on localization studies, for an ER-related function of the Nox complex. We present the first evidence that some functions of the BcNoxA complex are indeed linked to the ER, while others clearly require export from the ER. PMID:25402961

  1. Alkamides Activate Jasmonic Acid Biosynthesis and Signaling Pathways and Confer Resistance to Botrytis cinerea in Arabidopsis thaliana

    PubMed Central

    Méndez-Bravo, Alfonso; Calderón-Vázquez, Carlos; Ibarra-Laclette, Enrique; Raya-González, Javier; Ramírez-Chávez, Enrique; Molina-Torres, Jorge; Guevara-García, Angel A.; López-Bucio, José; Herrera-Estrella, Luis

    2011-01-01

    Alkamides are fatty acid amides of wide distribution in plants, structurally related to N-acyl-L-homoserine lactones (AHLs) from Gram-negative bacteria and to N- acylethanolamines (NAEs) from plants and mammals. Global analysis of gene expression changes in Arabidopsis thaliana in response to N-isobutyl decanamide, the most highly active alkamide identified to date, revealed an overrepresentation of defense-responsive transcriptional networks. In particular, genes encoding enzymes for jasmonic acid (JA) biosynthesis increased their expression, which occurred in parallel with JA, nitric oxide (NO) and H2O2 accumulation. The activity of the alkamide to confer resistance against the necrotizing fungus Botrytis cinerea was tested by inoculating Arabidopsis detached leaves with conidiospores and evaluating disease symptoms and fungal proliferation. N-isobutyl decanamide application significantly reduced necrosis caused by the pathogen and inhibited fungal proliferation. Arabidopsis mutants jar1 and coi1 altered in JA signaling and a MAP kinase mutant (mpk6), unlike salicylic acid- (SA) related mutant eds16/sid2-1, were unable to defend from fungal attack even when N-isobutyl decanamide was supplied, indicating that alkamides could modulate some necrotrophic-associated defense responses through JA-dependent and MPK6-regulated signaling pathways. Our results suggest a role of alkamides in plant immunity induction. PMID:22076141

  2. The hrpN gene of Erwinia amylovora stimulates tobacco growth and enhances resistance to Botrytis cinerea.

    PubMed

    Jang, Young-Sun; Sohn, Soo-In; Wang, Myeong-Hyeon

    2006-02-01

    Erwinia amylovora is a member of the harpin proteins that induces pathogen resistance and hypersensitive cell death in plants. To obtain tobacco plants displaying a hypersensitive response, the hrpN gene from Erwinia amylovora was cloned into vector pMJC-GB under the control of the rice cytochrome promoter and transfected into tobacco. Southern hybridization with a hrpN probe revealed that the gene was present in one copy in the transgenic plants. In addition, hrpN transcripts could be detected in transgenic plants but not in wild-type tobacco. The wild type gave 75 products in RAPD analysis with 12 primers while the transgenic plants gave 73, suggesting that hrpN gene had been integrated into the transgenic plant genomic DNA. The distribution of cell cycle phases in the wild type and transgenic plants was G0-G1: 71.25%, G2-M: 20.41%, S: 8.33%, while in transgenic plant was G0-G1: 54.95%, G2-M: 43.82%, S: 10.23%. The sizes of stomata and guard cells on transgenic leaves were similar to those of the wild type, but the epidermal cells were clearly smaller. The transgenic plants showed accelerated growth and development as well as enhanced resistance to Botrytis cinerea. PMID:16136336

  3. Solidago canadensis L. Essential Oil Vapor Effectively Inhibits Botrytis cinerea Growth and Preserves Postharvest Quality of Strawberry as a Food Model System

    PubMed Central

    Liu, Shumin; Shao, Xingfeng; Wei, Yanzhen; Li, Yonghua; Xu, Feng; Wang, Hongfei

    2016-01-01

    This study investigated the anti-fungal properties of Solidago canadensis L. essential oil (SCLEO) against Botrytis cinerea in vitro, and its ability to control gray mold and maintain quality in strawberry fruits. SCLEO exhibited dose-dependent antifungal activity against B. cinerea and profoundly altered mycelial morphology, cellular ultrastructure, and membrane permeability as evaluated by scanning electron microscopy, transmission electron microscopy, and fluorescence microscopy. SCLEO vapor at 0.1 mL/L maintained higher sensory acceptance and reduced decay of fresh strawberry fruit, and also reduced gray mold in artificially inoculated fruit. SCLEO treatment did not, however, stimulate phenylalanin ammonia-lyase, polyphenol oxidase, or chitinase, enzymes related to disease resistance. This suggests that SCLEO reduces gray mold by direct inhibition of pathogen growth. SCLEO vapor may provide a new and effective strategy for controlling postharvest disease and maintaining quality in strawberries. PMID:27531994

  4. Solidago canadensis L. Essential Oil Vapor Effectively Inhibits Botrytis cinerea Growth and Preserves Postharvest Quality of Strawberry as a Food Model System.

    PubMed

    Liu, Shumin; Shao, Xingfeng; Wei, Yanzhen; Li, Yonghua; Xu, Feng; Wang, Hongfei

    2016-01-01

    This study investigated the anti-fungal properties of Solidago canadensis L. essential oil (SCLEO) against Botrytis cinerea in vitro, and its ability to control gray mold and maintain quality in strawberry fruits. SCLEO exhibited dose-dependent antifungal activity against B. cinerea and profoundly altered mycelial morphology, cellular ultrastructure, and membrane permeability as evaluated by scanning electron microscopy, transmission electron microscopy, and fluorescence microscopy. SCLEO vapor at 0.1 mL/L maintained higher sensory acceptance and reduced decay of fresh strawberry fruit, and also reduced gray mold in artificially inoculated fruit. SCLEO treatment did not, however, stimulate phenylalanin ammonia-lyase, polyphenol oxidase, or chitinase, enzymes related to disease resistance. This suggests that SCLEO reduces gray mold by direct inhibition of pathogen growth. SCLEO vapor may provide a new and effective strategy for controlling postharvest disease and maintaining quality in strawberries. PMID:27531994

  5. Genome-wide analysis of pectate-induced gene expression in Botrytis cinerea: identification and functional analysis of putative d-galacturonate transporters.

    PubMed

    Zhang, Lisha; Hua, Chenlei; Stassen, Joost H M; Chatterjee, Sayantani; Cornelissen, Maxim; van Kan, Jan A L

    2014-11-01

    The fungal plant pathogen Botrytis cinerea produces a spectrum of cell wall degrading enzymes for the decomposition of host cell wall polysaccharides and the consumption of the monosaccharides that are released. Especially pectin is an abundant cell wall component, and the decomposition of pectin by B. cinerea has been extensively studied. An effective concerted action of the appropriate pectin depolymerising enzymes, monosaccharide transporters and catabolic enzymes is important for complete d-galacturonic acid utilization by B. cinerea. In this study, we performed RNA sequencing to compare genome-wide transcriptional profiles between B. cinerea cultures grown in media containing pectate or glucose as sole carbon source. Transcript levels of 32 genes that are induced by pectate were further examined in cultures grown on six different monosaccharides, by means of quantitative RT-PCR, leading to the identification of 8 genes that are exclusively induced by d-galacturonic acid. Among these, the hexose transporter encoding genes Bchxt15 and Bchxt19 were functionally characterised. The subcellular location was studied of BcHXT15-GFP and BcHXT19-GFP fusion proteins expressed under control of their native promoter, in a B. cinerea wild-type strain. Both genes are expressed during growth on d-galacturonic acid and the fusion proteins are localized in plasma membranes and intracellular vesicles. Target gene knockout analysis revealed that BcHXT15 contributes to d-galacturonic acid uptake at pH 5∼5.6. The virulence of all B. cinerea hexose transporter mutants tested was unaltered on tomato and Nicotiana benthamiana leaves. PMID:24140151

  6. Inadvertent gene silencing of argininosuccinate synthase (bcass1) in Botrytis cinerea by the pLOB1 vector system.

    PubMed

    Patel, Risha M; Van Kan, Jan A L; Bailey, Andy M; Foster, Gary D

    2010-09-01

    For several years, researchers working on the plant pathogen Botrytis cinerea and a number of other related fungi have routinely used the pLOB1 vector system, based on hygromycin resistance, under the control of the Aspergillus nidulans oliC promoter and what was reported to be the beta-tubulin (tubA) terminator. Recently, it has been demonstrated that this vector contains a 446-bp portion of the B. cinerea argininosuccinate synthase gene (bcass1) rather than the tubA terminator. As argininosuccinate synthase is essential for the production of L-arginine, inadvertent gene silencing of bcass1 may result in partial L-arginine auxotrophy and, indeed, may lead to altered phenotypes in planta. In this article, we report our findings relating to possible problems arising from this incorrect plasmid construction. As an absolute baseline, gene disruption of bcass1 was carried out and generated a strict auxotroph, unable to grow without exogenous arginine supplementation. The knockout displayed an alteration in host range in planta, showing a reduction in pathogenicity on strawberries, French bean leaves and tomatoes, but maintained wild-type growth on grape, which is in accordance with the reported arginine availability in such tissues. Deliberate gene silencing of bcass1 mirrored these effects, with strongly silenced lines showing reduced virulence. The degree of silencing as seen by partial auxotrophy was correlated with an observed reduction in virulence. We also showed that inadvertent silencing of bcass1 is possible when using the pLOB1 vector or derivatives thereof. Partial arginine auxotrophy and concomitant reductions in virulence were triggered in approximately 6% of transformants obtained when expressing enhanced green fluorescent protein, luciferase, monomeric red fluorescent protein or beta-glucuronidase using the pLOB1-based expression system, which inadvertently contains 446 bp of the bcass1 coding sequence. We recommend the testing of transformants obtained

  7. Analysis of the Molecular Dialogue Between Gray Mold (Botrytis cinerea) and Grapevine (Vitis vinifera) Reveals a Clear Shift in Defense Mechanisms During Berry Ripening.

    PubMed

    Kelloniemi, Jani; Trouvelot, Sophie; Héloir, Marie-Claire; Simon, Adeline; Dalmais, Bérengère; Frettinger, Patrick; Cimerman, Agnès; Fermaud, Marc; Roudet, Jean; Baulande, Sylvain; Bruel, Christophe; Choquer, Mathias; Couvelard, Linhdavanh; Duthieuw, Mathilde; Ferrarini, Alberto; Flors, Victor; Le Pêcheur, Pascal; Loisel, Elise; Morgant, Guillaume; Poussereau, Nathalie; Pradier, Jean-Marc; Rascle, Christine; Trdá, Lucie; Poinssot, Benoit; Viaud, Muriel

    2015-11-01

    Mature grapevine berries at the harvesting stage (MB) are very susceptible to the gray mold fungus Botrytis cinerea, while veraison berries (VB) are not. We conducted simultaneous microscopic and transcriptomic analyses of the pathogen and the host to investigate the infection process developed by B. cinerea on MB versus VB, and the plant defense mechanisms deployed to stop the fungus spreading. On the pathogen side, our genome-wide transcriptomic data revealed that B. cinerea genes upregulated during infection of MB are enriched in functional categories related to necrotrophy, such as degradation of the plant cell wall, proteolysis, membrane transport, reactive oxygen species (ROS) generation, and detoxification. Quantitative-polymerase chain reaction on a set of representative genes related to virulence and microscopic observations further demonstrated that the infection is also initiated on VB but is stopped at the penetration stage. On the plant side, genome-wide transcriptomic analysis and metabolic data revealed a defense pathway switch during berry ripening. In response to B. cinerea inoculation, VB activated a burst of ROS, the salicylate-dependent defense pathway, the synthesis of the resveratrol phytoalexin, and cell-wall strengthening. On the contrary, in infected MB, the jasmonate-dependent pathway was activated, which did not stop the fungal necrotrophic process. PMID:26267356

  8. The Transcription Factor BcLTF1 Regulates Virulence and Light Responses in the Necrotrophic Plant Pathogen Botrytis cinerea

    PubMed Central

    Schumacher, Julia; Simon, Adeline; Cohrs, Kim Christopher; Viaud, Muriel; Tudzynski, Paul

    2014-01-01

    Botrytis cinerea is the causal agent of gray mold diseases in a range of dicotyledonous plant species. The fungus can reproduce asexually by forming macroconidia for dispersal and sclerotia for survival; the latter also participate in sexual reproduction by bearing the apothecia after fertilization by microconidia. Light induces the differentiation of conidia and apothecia, while sclerotia are exclusively formed in the absence of light. The relevance of light for virulence of the fungus is not obvious, but infections are observed under natural illumination as well as in constant darkness. By a random mutagenesis approach, we identified a novel virulence-related gene encoding a GATA transcription factor (BcLTF1 for light-responsive TF1) with characterized homologues in Aspergillus nidulans (NsdD) and Neurospora crassa (SUB-1). By deletion and over-expression of bcltf1, we confirmed the predicted role of the transcription factor in virulence, and discovered furthermore its functions in regulation of light-dependent differentiation, the equilibrium between production and scavenging of reactive oxygen species (ROS), and secondary metabolism. Microarray analyses revealed 293 light-responsive genes, and that the expression levels of the majority of these genes (66%) are modulated by BcLTF1. In addition, the deletion of bcltf1 affects the expression of 1,539 genes irrespective of the light conditions, including the overexpression of known and so far uncharacterized secondary metabolism-related genes. Increased expression of genes encoding alternative respiration enzymes, such as the alternative oxidase (AOX), suggest a mitochondrial dysfunction in the absence of bcltf1. The hypersensitivity of Δbctlf1 mutants to exogenously applied oxidative stress - even in the absence of light - and the restoration of virulence and growth rates in continuous light by antioxidants, indicate that BcLTF1 is required to cope with oxidative stress that is caused either by exposure to light

  9. UVR8 mediates UV-B-induced Arabidopsis defense responses against Botrytis cinerea by controlling sinapate accumulation.

    PubMed

    Demkura, Patricia V; Ballaré, Carlos L

    2012-05-01

    Light is emerging as a central regulator of plant immune responses against herbivores and pathogens. Solar UV-B radiation plays an important role as a positive modulator of plant defense. However, since UV-B photons can interact with a wide spectrum of molecular targets in plant tissues, the mechanisms that mediate their effects on plant defense have remained elusive. Here, we show that ecologically meaningful doses of UV-B radiation increase Arabidopsis resistance to the necrotrophic fungus Botrytis cinerea and that this effect is mediated by the photoreceptor UVR8. The UV-B effect on plant resistance was conserved in mutants impaired in jasmonate (JA) signaling (jar1-1 and P35S:JAZ10.4) or metabolism of tryptophan-derived defense compounds (pen2-1, pad3-1, pen2 pad3), suggesting that neither regulation of the JA pathway nor changes in levels of indolic glucosinolates (iGS) or camalexin are involved in this response. UV-B radiation, acting through UVR8, increased the levels of flavonoids and sinapates in leaf tissue. The UV-B effect on pathogen resistance was still detectable in tt4-1, a mutant deficient in chalcone synthase and therefore impaired in the synthesis of flavonoids, but was absent in fah1-7, a mutant deficient in ferulic acid 5-hydroxylase, which is essential for sinapate biosynthesis. Collectively, these results indicate that UVR8 plays an important role in mediating the effects of UV-B radiation on pathogen resistance by controlling the expression of the sinapate biosynthetic pathway. PMID:22447155

  10. Effects of Two Salts Compounds on Mycelial Growth, Sporulation, and Spore Germination of Six Isolates of Botrytis cinerea in the Western North of Algeria.

    PubMed

    Boumaaza, Boualem; Benkhelifa, Mohamed; Belkhoudja, Moulay

    2015-01-01

    Six isolates of Botrytis cinerea were isolated from leaves and stems of different tomato varieties taken from four areas in the northwest of Algeria where tomato is mostly grown in greenhouses and high tunnels. The purpose of this research was to determine the effect of two salts, NaCl and CaCl2, on three stages of Botrytis cinerea's life cycle. All isolates tested were stimulated in 50 to 150 ppm; NaCl was the most effective treatment to increase mycelial growth at two tested concentrations. However, at 300 ppm concentration, CaCl2 completely inhibited the growth of mycelium; they reach 34.78% for the isolate TR46 and 26.72% for isolate F27. The sodium and calcium salts stimulated conidia production in liquid culture. We noticed that the effect of calcium chloride on sporulation was average while sodium chloride. In the medium containing 50 ppm, calcium chloride and sodium chloride increased the germination capacity of most isolates compared with the control. Other calcium salts, at 100 or 300 ppm, decreased the germination percentage of the conidia. With the exception of sodium salts, the inhibitions of germination reduce at 150 or 300 compared with the control. Conidial germination was slightly inhibited by sodium chloride only when the concentration was over 300 ppm. PMID:25883657

  11. Effects of Two Salts Compounds on Mycelial Growth, Sporulation, and Spore Germination of Six Isolates of Botrytis cinerea in the Western North of Algeria

    PubMed Central

    Boumaaza, Boualem; Benkhelifa, Mohamed; Belkhoudja, Moulay

    2015-01-01

    Six isolates of Botrytis cinerea were isolated from leaves and stems of different tomato varieties taken from four areas in the northwest of Algeria where tomato is mostly grown in greenhouses and high tunnels. The purpose of this research was to determine the effect of two salts, NaCl and CaCl2, on three stages of Botrytis cinerea's life cycle. All isolates tested were stimulated in 50 to 150 ppm; NaCl was the most effective treatment to increase mycelial growth at two tested concentrations. However, at 300 ppm concentration, CaCl2 completely inhibited the growth of mycelium; they reach 34.78% for the isolate TR46 and 26.72% for isolate F27. The sodium and calcium salts stimulated conidia production in liquid culture. We noticed that the effect of calcium chloride on sporulation was average while sodium chloride. In the medium containing 50 ppm, calcium chloride and sodium chloride increased the germination capacity of most isolates compared with the control. Other calcium salts, at 100 or 300 ppm, decreased the germination percentage of the conidia. With the exception of sodium salts, the inhibitions of germination reduce at 150 or 300 compared with the control. Conidial germination was slightly inhibited by sodium chloride only when the concentration was over 300 ppm. PMID:25883657

  12. Involvement of protein tyrosine phosphatases BcPtpA and BcPtpB in regulation of vegetative development, virulence and multi-stress tolerance in Botrytis cinerea.

    PubMed

    Yang, Qianqian; Yu, Fangwei; Yin, Yanni; Ma, Zhonghua

    2013-01-01

    Tyrosine phosphorylation and dephosphorylation have emerged as fundamentally important mechanisms of signal transduction and regulation in eukaryotic cells, governing many processes, but little has been known about their functions in filamentous fungi. In this study, we deleted two putative protein tyrosine phosphatase (PTP) genes (BcPTPA and BcPTPB) in Botrytis cinerea, encoding the orthologs of Saccharomyces cerevisiae Ptp2 and Ptp3, respectively. Although BcPtpA and BcPtpB have opposite functions in conidiation, they are essential for sclerotial formation in B. cinerea. BcPTPA and BcPTPB deletion mutants ΔBcPtpA-10 and ΔBcPtpB-4 showed significantly increased sensitivity to osmotic and oxidative stresses, and to cell wall damaging agents. Inoculation tests showed that both mutants exhibited dramatically decreased virulence on tomato leaves, apples and grapes. In S. cerevisiae, it has been shown that Ptp2 and Ptp3 negatively regulate the high-osmolarity glycerol (HOG) pathway and the cell wall integrity (CWI) pathway. Although both BcPtpA and BcPtpB were able to inactive Hog1 and Mpk1 in S. cerevisiae, in contrast to S. cerevisiae, they positively regulate phosphorylation of BcSak1 (the homologue of Hog1) and BcBmp3 (the homologue of Mpk1) in B. cinerea under stress conditions. These results demonstrated that functions of PTPs in B. cinerea are different from those in S. cerevisiae, and BcPtpA and BcPtpB play important roles in regulation of vegetative development, virulence and in adaptation to oxidative, osmotic and cell-wall damage stresses in B. cinerea. PMID:23585890

  13. Exploring Mechanisms of Resistance to Respiratory Inhibitors in Field Strains of Botrytis cinerea, the Causal Agent of Gray Mold ▿ †

    PubMed Central

    Leroux, Pierre; Gredt, Michel; Leroch, Michaela; Walker, Anne-Sophie

    2010-01-01

    Respiratory inhibitors are among the fungicides most widely used for disease control on crops. Most are strobilurins and carboxamides, inhibiting the cytochrome b of mitochondrial complex III and the succinate dehydrogenase of mitochondrial complex II, respectively. A few years after the approval of these inhibitors for use on grapevines, field isolates of Botrytis cinerea, the causal agent of gray mold, resistant to one or both of these classes of fungicide were recovered in France and Germany. However, little was known about the mechanisms underlying this resistance in field populations of this fungus. Such knowledge could facilitate resistance risk assessment. The aim of this study was to investigate the mechanisms of resistance occurring in B. cinerea populations. Highly specific resistance to strobilurins was correlated with a single mutation of the cytb target gene. Changes in its intronic structure may also have occurred due to an evolutionary process controlling selection for resistance. Specific resistance to carboxamides was identified for six phenotypes, with various patterns of resistance levels and cross-resistance. Several mutations specific to B. cinerea were identified within the sdhB and sdhD genes encoding the iron-sulfur protein and an anchor protein of the succinate dehydrogenase complex. Another as-yet-uncharacterized mechanism of resistance was also recorded. In addition to target site resistance mechanisms, multidrug resistance, linked to the overexpression of membrane transporters, was identified in strains with low to moderate resistance to several respiratory inhibitors. This diversity of resistance mechanisms makes resistance management difficult and must be taken into account when developing strategies for Botrytis control. PMID:20693447

  14. Ectopic expression of a grape aspartic protease gene, AP13, in Arabidopsis thaliana improves resistance to powdery mildew but increases susceptibility to Botrytis cinerea.

    PubMed

    Guo, Rongrong; Tu, Mingxing; Wang, Xianhang; Zhao, Jiao; Wan, Ran; Li, Zhi; Wang, Yuejin; Wang, Xiping

    2016-07-01

    The grape aspartic protease gene, AP13 was previously reported to be responsive, in Chinese wild Vitis quinquangularis cv. 'Shang-24', to infection by Erysiphe necator, the causal agent of powdery mildew disease, as well as to treatment with salicylic acid in V. labrusca×V. vinifera cv. 'Kyoho'. In the current study, we evaluated the expression levels of AP13 in 'Shang-24' in response to salicylic acid (SA), methyl jasmonate (MeJA) and ethylene (ET) treatments, as well as to infection by the necrotrophic fungus, Botrytis cinerea, and the transcript levels of VqAP13 decreased after B. cinerea infection and MeJA treatment, but increased following ET and SA treatments. Transgenic Arabidopsis thaliana lines over-expressing VqAP13 under the control of a constitutive promoter showed enhanced resistance to powdery mildew and to the bacterium Pseudomonas syringae pv. tomato DC3000, and accumulated more callose than wild type plants, while the resistance of transgenic A. thaliana lines to B. cinerea inoculation was reduced. In addition, the expression profiles of various disease resistance- related genes in the transgenic A. thaliana lines following infection by different pathogens were compared to the equivalent profiles in the wild type plants. The results suggest that VqAP13 action promotes the SA dependent signal transduction pathway, but suppresses the JA signal transduction pathway. PMID:27181943

  15. DHN melanin biosynthesis in the plant pathogenic fungus Botrytis cinerea is based on two developmentally regulated key enzyme (PKS)-encoding genes.

    PubMed

    Schumacher, Julia

    2016-02-01

    Botrytis cinerea is the causal agent of gray mold disease in various plant species and produces grayish macroconidia and/or black sclerotia at the end of the infection cycle. It has been suggested that the pigmentation is due to the accumulation of 1,8-dihydroxynaphthalene (DHN) melanin. To unravel its basis and regulation, the putative melanogenic and regulatory genes were identified and functionally characterized. Unlike other DHN melanin-producing fungi, B. cinerea and other Leotiomycetes contain two key enzyme (PKS)-encoding enzymes. Bcpks12 and bcpks13 are developmentally regulated and are required for melanogenesis in sclerotia and conidia respectively. BcYGH1 converts the BcPKS13 product and contributes thereby to conidial melanogenesis. In contrast, enzymes acting downstream in conversion of the PKS products (BcBRN2, BcSCD1 and BcBRN1) are required for both, sclerotial and conidial melanogenesis, suggesting that DHN melanogenesis in B. cinerea follows a non-linear pathway that is rather unusual for secondary metabolic pathways. Regulation of the melanogenic genes involves three pathway-specific transcription factors (TFs) that are clustered with bcpks12 or bcpks13 and other developmental regulators such as light-responsive TFs. Melanogenic genes are dispensable in vegetative mycelia for proper growth and virulence. However, DHN melanin is considered to contribute to the longevity of the reproduction structures. PMID:26514268

  16. Fungistatic activity of Zanthoxylum rhoifolium Lam. bark extracts against fungal plant pathogens and investigation on mechanism of action in Botrytis cinerea.

    PubMed

    Carotenuto, Gennaro; Carrieri, Raffaele; Tarantino, Paola; Alfieri, Mariaevelina; Leone, Antonella; De Tommasi, Nunziatina; Lahoz, Ernesto

    2015-01-01

    Plant-derived compounds are emerging as an alternative choice to synthetic fungicides. Chloroform-methanol extract, obtained from the bark of Zanthoxylum rhoifolium, a member of Rutaceae, showed a fungistatic effect on Botrytis cinerea, Sclerotinia sclerotiorum, Alternaria alternata, Colletotrichum gloeosporioides and Clonostachys rosea, when added to the growth medium at different concentrations. A fraction obtained by gel separation and containing the alkaloid O-Methylcapaurine showed significant fungistatic effect against B. cinerea and S. sclerotiorum, two of the most destructive phytopathogenic fungi. The underlying mechanism of such an inhibition was further investigated in B. cinerea, a fungus highly prone to develop fungicide resistance, by analysing the expression levels of a set of genes (BcatrB, P450, CYP51 and TOR). O-Methylcapaurine inhibited the expression of all the analysed genes. In particular, the expression of BcatrB gene, encoding a membrane drug transporter involved in the resistance to a wide range of xenobiotic compounds, was strongly inhibited (91%). PMID:25589008

  17. CYP94-mediated jasmonoyl-isoleucine hormone oxidation shapes jasmonate profiles and attenuates defence responses to Botrytis cinerea infection

    PubMed Central

    Aubert, Yann; Widemann, Emilie; Miesch, Laurence; Pinot, Franck; Heitz, Thierry

    2015-01-01

    Induced resistance to the necrotrophic pathogen Botrytis cinerea depends on jasmonate metabolism and signalling in Arabidopsis. We have presented here extensive jasmonate profiling in this pathosystem and investigated the impact of the recently reported jasmonoyl-isoleucine (JA-Ile) catabolic pathway mediated by cytochrome P450 (CYP94) enzymes. Using a series of mutant and overexpressing (OE) plant lines, we showed that CYP94B3 and CYP94C1 are integral components of the fungus-induced jasmonate metabolic pathway and control the abundance of oxidized conjugated but also some unconjugated derivatives, such as sulfated 12-HSO4-JA. Despite causing JA-Ile overaccumulation due to impaired oxidation, CYP94 deficiency had negligible impacts on resistance, associated with enhanced JAZ repressor transcript levels. In contrast, plants overexpressing (OE) CYP94B3 or CYP94C1 were enriched in 12-OH-JA-Ile or 12-COOH-JA-Ile respectively. This shift towards oxidized JA-Ile derivatives was concomitant with strongly impaired defence gene induction and reduced disease resistance. CYP94B3-OE, but unexpectedly not CYP94C1-OE, plants displayed reduced JA-Ile levels compared with the wild type, suggesting that increased susceptibility in CYP94C1-OE plants may result from changes in the hormone oxidation ratio rather than absolute changes in JA-Ile levels. Consistently, while feeding JA-Ile to seedlings triggered strong induction of JA pathway genes, induction was largely reduced or abolished after feeding with the CYP94 products 12-OH-JA-Ile and 12-COOH-JA-Ile, respectively. This trend paralleled in vitro pull-down assays where 12-COOH-JA-Ile was unable to promote COI1–JAZ9 co-receptor assembly. Our results highlight the dual function of CYP94B3/C1 in antimicrobial defence: by controlling hormone oxidation status for signal attenuation, these enzymes also define JA-Ile as a metabolic hub directing jasmonate profile complexity. PMID:25903915

  18. CYP94-mediated jasmonoyl-isoleucine hormone oxidation shapes jasmonate profiles and attenuates defence responses to Botrytis cinerea infection.

    PubMed

    Aubert, Yann; Widemann, Emilie; Miesch, Laurence; Pinot, Franck; Heitz, Thierry

    2015-07-01

    Induced resistance to the necrotrophic pathogen Botrytis cinerea depends on jasmonate metabolism and signalling in Arabidopsis. We have presented here extensive jasmonate profiling in this pathosystem and investigated the impact of the recently reported jasmonoyl-isoleucine (JA-Ile) catabolic pathway mediated by cytochrome P450 (CYP94) enzymes. Using a series of mutant and overexpressing (OE) plant lines, we showed that CYP94B3 and CYP94C1 are integral components of the fungus-induced jasmonate metabolic pathway and control the abundance of oxidized conjugated but also some unconjugated derivatives, such as sulfated 12-HSO4-JA. Despite causing JA-Ile overaccumulation due to impaired oxidation, CYP94 deficiency had negligible impacts on resistance, associated with enhanced JAZ repressor transcript levels. In contrast, plants overexpressing (OE) CYP94B3 or CYP94C1 were enriched in 12-OH-JA-Ile or 12-COOH-JA-Ile respectively. This shift towards oxidized JA-Ile derivatives was concomitant with strongly impaired defence gene induction and reduced disease resistance. CYP94B3-OE, but unexpectedly not CYP94C1-OE, plants displayed reduced JA-Ile levels compared with the wild type, suggesting that increased susceptibility in CYP94C1-OE plants may result from changes in the hormone oxidation ratio rather than absolute changes in JA-Ile levels. Consistently, while feeding JA-Ile to seedlings triggered strong induction of JA pathway genes, induction was largely reduced or abolished after feeding with the CYP94 products 12-OH-JA-Ile and 12-COOH-JA-Ile, respectively. This trend paralleled in vitro pull-down assays where 12-COOH-JA-Ile was unable to promote COI1-JAZ9 co-receptor assembly. Our results highlight the dual function of CYP94B3/C1 in antimicrobial defence: by controlling hormone oxidation status for signal attenuation, these enzymes also define JA-Ile as a metabolic hub directing jasmonate profile complexity. PMID:25903915

  19. Fungicide resistance of Botrytis cinerea in tomato greenhouses in the Canary Islands and effectiveness of non-chemical treatments against gray mold.

    PubMed

    Rodríguez, A; Acosta, A; Rodríguez, C

    2014-09-01

    Tomato greenhouses in the Canary Islands, Spain, were surveyed to estimate frequencies of resistance to benzimidazoles, dicarboximides, anilinopyrimidines and N-phenylcarbamates in Botrytis cinerea. Resistance to carbendazim, iprodione, pyrimethanil and diethofencarb was found in 74.2, 86.4, 28.8 and 31.8% of isolates, respectively. Benzimidazole- and anilinopyrimide-resistant isolates were highly resistant, showing EC50 values above 500 µg/ml carbendazim and a mean EC50 value of 28.42 µg/ml pyrimethanil, respectively. By contrast, a low level of resistance was observed among dicarboximide-resistant isolates (mean EC50 value of 1.81 µg/ml iprodione). Phenotypes with double resistance to carbendazim and iprodione, and triple resistance to carbendazim, iprodione and pyrimethanil were the most common, occurring in 36.4 and 28.8% of isolates. The surveyed greenhouses had never been treated with fenhexamid and Signum™ (pre-packed mixture of boscalid and pyraclostrobin), and baseline sensitivities of B. cinerea isolates to these fungicides were determined. The EC50 values were within the range of 0.009-0.795 µg/ml fenhexamid and of 0.014-0.48 µg/ml Signum. In addition, available formulations based on elicitors of plant defense response and biocontrol agents were evaluated against B. cinerea in tomato plants under semi-controlled greenhouse conditions, the yeast Candida sake CPA-1 being able to reduce gray mold significantly when it was applied on petiole wounds and the plants were inoculated 24 h later. Likewise, C. sake was effective against B. cinerea in harvested tomato fruits, yeast-treated tomatoes showed a 70.66 and 30.31% reduction in the diameters of decay lesions compared with controls after 10 days of storage at 20 and 9 °C, respectively. PMID:24817605

  20. Induction of Direct or Priming Resistance against Botrytis cinerea in Strawberries by β-Aminobutyric Acid and Their Effects on Sucrose Metabolism.

    PubMed

    Wang, Kaituo; Liao, Yunxia; Xiong, Qi; Kan, Jianquan; Cao, Shifeng; Zheng, Yonghua

    2016-07-27

    The specific forms of disease resistance induced by β-aminobutyric acid (BABA) and their impacts on sucrose metabolism of postharvest strawberries were determined in the present research. Treatment with 10-500 mmol L(-1) BABA inhibited the Botrytis cinerea infection, possibly directly by suppressing the fungus growth and indirectly by triggering disease resistance. Moreover, BABA-induced resistance against B. cinerea infection in strawberries was associated with either one of two mechanisms, depending upon the concentration used: BABA at concentrations higher than 100 mmol L(-1) directly induced the defense response, including a H2O2 burst, modulation of the expression of PR genes, including FaPR1, FaChi3, Faβglu, and FaPAL, and increased activities of chitinase, β-1,3-glucanase, and PAL, whereas BABA at 10 mmol L(-1) activated a priming response because the BABA-treated fruits exhibited an increased capacity to express molecular defense only when the fruits were inoculated with B. cinerea. Activation of the priming defense appeared almost as effective against B. cinerea as inducing direct defense. However, the primed strawberries maintained higher activities of SS synthesis and SPS and SPP enzymes) and lower level of SS cleavage during the incubation; these activities contributed to higher sucrose, fructose, and glucose contents, sweetness index, and sensory scores compared to fruits exhibiting the direct defense. Thus, it is plausible that the priming defense, which can be activated by BABA at relatively low concentrations, represents an optimal strategy for combining the advantages of enhanced disease protection and soluble sugar accumulation. PMID:27368357

  1. Absence of the endo-beta-1,4-glucanases Cel1 and Cel2 reduces susceptibility to Botrytis cinerea in tomato.

    PubMed

    Flors, Víctor; Leyva, María de la O; Vicedo, Begonya; Finiti, Ivan; Real, María Dolores; García-Agustín, Pilar; Bennett, Alan B; González-Bosch, Carmen

    2007-12-01

    Cel1 and Cel2 are members of the tomato (Solanum lycopersicum Mill) endo-beta-1,4-glucanase (EGase) family that may play a role in fruit ripening and organ abscission. This work demonstrates that Cel1 protein is present in other vegetative tissues and accumulates during leaf development. We recently reported the downregulation of both the Cel1 mRNA and protein upon fungal infection, suggesting the involvement of EGases in plant-pathogen interactions. This hypothesis was confirmed by assessing the resistance to Botrytis cinerea infection of transgenic plants expressing both genes in an antisense orientation (Anti-Cel1, Anti-Cel2 and Anti-Cel1-Cel2). The Anti-Cel1-Cel2 plants showed enhanced resistance to this fungal necrotroph. Microscopical analysis of infected leaves revealed that tomato plants accumulated pathogen-inducible callose within the expanding lesion. Anti-Cel1-Cel2 plants presented a faster and enhanced callose accumulation against B. cinerea than wild-type plants. The inhibitor 2-deoxy-d-glucose, a callose synthesis inhibitor, showed a direct relationship between faster callose accumulation and enhanced resistance to B. cinerea. EGase activity appears to negatively modulate callose deposition. The absence of both EGase genes was associated with changes in the expression of the pathogen-related genes PR1 and LoxD. Interestingly, Anti-Cel1-Cel2 plants were more susceptible to Pseudomonas syringae, displaying severe disease symptoms and enhanced bacterial growth relative to wild-type plants. Analysis of the involvement of Cel1 and Cel2 in the susceptibility to B. cinerea in fruits was done with the ripening-impaired mutants Never ripe (Nr) and Ripening inhibitor (rin). The data reported in this work support the idea that enzymes involved in cell wall metabolism play a role in susceptibility to pathogens. PMID:17916112

  2. Reduction of nectarine decay caused by Rhizopus stolonifer, Botrytis cinerea and Penicillium digitatum with Aloe vera gel alone or with the addition of thymol.

    PubMed

    Navarro, Diana; Díaz-Mula, Huertas M; Guillén, Fabián; Zapata, Pedro J; Castillo, Salvador; Serrano, María; Valero, Daniel; Martínez-Romero, Domingo

    2011-12-01

    Two nectarine cultivars ('Flavela' and 'Flanoba') were treated with Aloe vera gel alone, or with the addition of thymol, and then inoculated with Rhizopus stolonifer, Botrytis cinerea and Penicillium digitatum. Both treatments were effective in reducing the decay incidence caused by the 3 fungi species, although the addition of thymol did not generally improve the efficacy of Aloe vera gel on reducing the infection damage. The coatings were clearly effective in reducing the postharvest ripening process of both nectarine cultivars manifested by a delay in ethylene production and respiration rate, weight loss and softening. Interestingly, these coatings showed effectiveness on reducing decay development in inoculated fruits and thus Aloe vera could be considered as natural antifungal compound and might serve as alternative of synthetic fungicides. PMID:21974979

  3. Burkholderia phytofirmans PsJN Confers Grapevine Resistance against Botrytis cinerea via a Direct Antimicrobial Effect Combined with a Better Resource Mobilization

    PubMed Central

    Miotto-Vilanova, Lidiane; Jacquard, Cédric; Courteaux, Barbara; Wortham, Laurence; Michel, Jean; Clément, Christophe; Barka, Essaïd A.; Sanchez, Lisa

    2016-01-01

    Plant innate immunity serves as a surveillance system by providing the first line of powerful weapons to fight against pathogen attacks. Beneficial microorganisms and Microbial-Associated Molecular Patterns might act as signals to trigger this immunity. Burkholderia phytofirmans PsJN, a highly efficient plant beneficial endophytic bacterium, promotes growth in a wide variety of plants including grapevine. Further, the bacterium induces plant resistance against abiotic and biotic stresses. However, no study has deciphered triggered-mechanisms during the tripartite interaction between grapevine, B. phytofirmans PsJN and Botrytis cinerea. Herein, we showed that in contrast with classical rhizobacteria, which are restricted in the root system and act through ISR, B. phytofirmans PsJN is able to migrate until aerial part and forms at leaves surface a biofilm around B. cinerea mycelium to restrict the pathogen. Nevertheless, considering the endophytic level of PsJN in leaves, the plant protection efficacy of B. phytofirmans PsJN could not be explained solely by its direct antifungal effect. Deeper investigations showed a callose deposition, H2O2 production and primed expression of PR1, PR2, PR5, and JAZ only in bacterized-plantlets after pathogen challenge. The presence of PsJN modulated changes in leaf carbohydrate metabolism including gene expression, sugar levels, and chlorophyll fluorescence imaging after Botrytis challenge. Our findings indicated that protection induced by B. phytofirmans PsJN was multifaceted and relied on a direct antifungal effect, priming of defense mechanisms as well as the mobilization of carbon sources in grapevine leaf tissues. PMID:27602036

  4. Burkholderia phytofirmans PsJN Confers Grapevine Resistance against Botrytis cinerea via a Direct Antimicrobial Effect Combined with a Better Resource Mobilization.

    PubMed

    Miotto-Vilanova, Lidiane; Jacquard, Cédric; Courteaux, Barbara; Wortham, Laurence; Michel, Jean; Clément, Christophe; Barka, Essaïd A; Sanchez, Lisa

    2016-01-01

    Plant innate immunity serves as a surveillance system by providing the first line of powerful weapons to fight against pathogen attacks. Beneficial microorganisms and Microbial-Associated Molecular Patterns might act as signals to trigger this immunity. Burkholderia phytofirmans PsJN, a highly efficient plant beneficial endophytic bacterium, promotes growth in a wide variety of plants including grapevine. Further, the bacterium induces plant resistance against abiotic and biotic stresses. However, no study has deciphered triggered-mechanisms during the tripartite interaction between grapevine, B. phytofirmans PsJN and Botrytis cinerea. Herein, we showed that in contrast with classical rhizobacteria, which are restricted in the root system and act through ISR, B. phytofirmans PsJN is able to migrate until aerial part and forms at leaves surface a biofilm around B. cinerea mycelium to restrict the pathogen. Nevertheless, considering the endophytic level of PsJN in leaves, the plant protection efficacy of B. phytofirmans PsJN could not be explained solely by its direct antifungal effect. Deeper investigations showed a callose deposition, H2O2 production and primed expression of PR1, PR2, PR5, and JAZ only in bacterized-plantlets after pathogen challenge. The presence of PsJN modulated changes in leaf carbohydrate metabolism including gene expression, sugar levels, and chlorophyll fluorescence imaging after Botrytis challenge. Our findings indicated that protection induced by B. phytofirmans PsJN was multifaceted and relied on a direct antifungal effect, priming of defense mechanisms as well as the mobilization of carbon sources in grapevine leaf tissues. PMID:27602036

  5. nor-Mevaldic acid surrogates as selective antifungal agent leads against Botrytis cinerea. Enantioselective preparation of 4-hydroxy-6-(1-phenylethoxy)tetrahydro-2H-pyran-2-one.

    PubMed

    Botubol-Ares, José Manuel; Durán-Peña, María Jesús; Hernández-Galán, Rosario; Collado, Isidro G; Harwood, Laurence M; Macías-Sánchez, Antonio J

    2015-07-01

    Solvent-free desymmetrisation of meso-dialdehyde 1 with chiral 1-phenylethan-1-ol, led to preparation of 4-silyloxy-6-alkyloxytetrahydro-2H-pyran-2-one (+)-3a with a 96:4 dr Deprotected lactone (+)-19a and the related racemic lactones 16a-18a present a lactone moiety resembling the natural substrate of HMG-CoA reductase and their antifungal properties have been evaluated against the phytopathogenic fungi Botrytis cinerea and Colletotrichum gloeosporioides. These compounds were selectively active against B. cinerea, while inactive against C. gloeosporioides. PMID:25971873

  6. Biocontrol ability and action mechanism of food-isolated yeast strains against Botrytis cinerea causing post-harvest bunch rot of table grape.

    PubMed

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2015-05-01

    Strains belonging to the species Saccharomyces cerevisiae, Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans, isolated from different food sources, were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic mold Botrytis cinerea. All yeast strains demonstrated antifungal activity at different levels depending on species and medium. Killer strains of W. anomalus and S. cerevisiae showed the highest biocontrol in vitro activity, as demonstrated by largest inhibition halos. The competition for iron and the ability to form biofilm and to colonize fruit wounds were hypothesized as the main action mechanisms for M. pulcherrima. The production of hydrolytic enzymes and the ability to colonize the wounds were the most important mechanisms for biocontrol activity in A. pullulans and W. anomalus, which also showed high ability to form biofilm. The production of volatile organic compounds (VOCs) with in vitro and in vivo inhibitory effect on pathogen growth was observed for the species W. anomalus, S. cerevisiae and M. pulcherrima. Our study clearly indicates that multiple modes of action may explain as M. pulcherrima provide excellent control of postharvest botrytis bunch rot of grape. PMID:25583341

  7. Epiphyas postvittana (Lepidoptera: Tortricidae)-Botrytis cinerea (Helotiales: Sclerotiniaceae)-Vitis vinifera (Vitales: Vitaceae) Interaction: The Role of B. cinerea on the Development of E. postvittana in Synthetic Nutritional Media.

    PubMed

    Rizvi, S Z M; Raman, A

    2015-08-01

    Epiphyas postvittana (Walker) (light-brown apple moth) is a polyphagous herbivore of economic significance, which also feeds on Vitis vinifera L. The E. postvittana-V. vinifera interacting system also involves the participation of the fungus Botrytis cinerea Persoon ex Fries. We have been exploring the relationship among E. postvittana-V. vinifera-B. cinerea over the past two years. In this article, we report the preference and performance of the larvae of E. postvittana raised solely on a synthetic diet incorporated with the mycelial material of B. cinerea (Diet B). To characterize the effect of fungus on the development of E. postvittana, another synthetic diet was prepared that included the lyophilized leaf material of V. vinifera (Diet C). When raised on Diets B and C, a decrease in the duration of larval development and an increase in the survival and fecundity rate of E. postvittana occurred. Diet B influenced the pupal mass, but a significant increase occurred when the larvae were fed on Diet C. The larval emergence rate was the greatest in E. postvittana raised on Diet B, followed by those on Diet C. The F(2) generation of the larvae reared on Diet B showed similar effects as F(1) on the life-history performance of the larvae. Diet B enhanced the life-history performance of E. postvittana, although the larvae of E. postvittana showed little preference to Diet B. The greater fertility rate of E. postvittana reared on Diet B suggests the importance of sterols as shown in Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae) and in a few Myrmicinae (Hymenoptera: Formicidae), which serve as precursors to different ecdysteroids that regulate many critical processes through embryonic development. PMID:26470305

  8. Involvement of salicylic acid, glutathione and protein S-thiolation in plant cell death-mediated defence response of Mesembryanthemum crystallinum against Botrytis cinerea.

    PubMed

    Kuźniak, Elżbieta; Kaźmierczak, Andrzej; Wielanek, Marzena; Głowacki, Rafał; Kornas, Andrzej

    2013-02-01

    The response of Mesembryanthemum crystallinum plants performing C3 photosynthesis and crassulacean acid metabolism (CAM) to the non-host necrotrophic pathogen Botrytis cinerea was analyzed at the local and systemic levels. The induction of programmed cell death, lignin and callose deposition, changes in salicylic acid, glutathione and cysteinylglycine pools as well as the content of thiolated proteins were studied. The infected C3 and CAM plants exhibited hypersensitive-like defence response, however fluorescence staining with acridine orange and ethidium bromide revealed programmed cell death events in C3 plants only. The local immune response was not related to callose and lignin deposition. In the infected plants, salicylic acid, glutathione and cysteinylglycine, the first product of glutathione catabolism, as well as protein S-thiolation, predominantly S-glutathionylation, contributed to local defence at sites of inoculation. They (except protein thiolation) were also active in the establishment of systemic acclimation response monitored in the non-treated upper leaves. The extent to which they were involved in the local and systemic responses induced by B. cinerea differed in C3 and CAM plants. The accumulation of free salicylic acid, both in treated and upper leaves of the infected plants, was much more pronounced in CAM plants. The results have been discussed with respect to redox regulations in defence against necrotrophic pathogens and to stress acclimation. PMID:23228550

  9. Abscisic Acid Deficiency Causes Changes in Cuticle Permeability and Pectin Composition That Influence Tomato Resistance to Botrytis cinerea1[C][W][OA

    PubMed Central

    Curvers, Katrien; Seifi, Hamed; Mouille, Grégory; de Rycke, Riet; Asselbergh, Bob; Van Hecke, Annelies; Vanderschaeghe, Dieter; Höfte, Herman; Callewaert, Nico; Van Breusegem, Frank; Höfte, Monica

    2010-01-01

    A mutant of tomato (Solanum lycopersicum) with reduced abscisic acid (ABA) production (sitiens) exhibits increased resistance to the necrotrophic fungus Botrytis cinerea. This resistance is correlated with a rapid and strong hydrogen peroxide-driven cell wall fortification response in epidermis cells that is absent in tomato with normal ABA production. Moreover, basal expression of defense genes is higher in the mutant compared with the wild-type tomato. Given the importance of this fast response in sitiens resistance, we investigated cell wall and cuticle properties of the mutant at the chemical, histological, and ultrastructural levels. We demonstrate that ABA deficiency in the mutant leads to increased cuticle permeability, which is positively correlated with disease resistance. Furthermore, perturbation of ABA levels affects pectin composition. sitiens plants have a relatively higher degree of pectin methylesterification and release different oligosaccharides upon inoculation with B. cinerea. These results show that endogenous plant ABA levels affect the composition of the tomato cuticle and cell wall and demonstrate the importance of cuticle and cell wall chemistry in shaping the outcome of this plant-fungus interaction. PMID:20709830

  10. Bcmfs1, a Novel Major Facilitator Superfamily Transporter from Botrytis cinerea, Provides Tolerance towards the Natural Toxic Compounds Camptothecin and Cercosporin and towards Fungicides

    PubMed Central

    Hayashi, Keisuke; Schoonbeek, Henk-jan; De Waard, Maarten A.

    2002-01-01

    Bcmfs1, a novel major facilitator superfamily gene from Botrytis cinerea, was cloned, and replacement and overexpression mutants were constructed to study its function. Replacement mutants showed increased sensitivity to the natural toxic compounds camptothecin and cercosporin, produced by the plant Camptotheca acuminata and the plant pathogenic fungus Cercospora kikuchii, respectively. Overexpression mutants displayed decreased sensitivity to these compounds and to structurally unrelated fungicides, such as sterol demethylation inhibitors (DMIs). A double-replacement mutant of Bcmfs1 and the ATP-binding cassette (ABC) transporter gene BcatrD was more sensitive to DMI fungicides than a single-replacement mutant of BcatrD, known to encode an important ABC transporter of DMIs. The sensitivity of the wild-type strain and mutants to DMI fungicides correlated with Bcmfs1 expression levels and with the initial accumulation of oxpoconazole by germlings of these isolates. The results indicate that Bcmfs1 is a major facilitator superfamily multidrug transporter involved in protection against natural toxins and fungicides and has a substrate specificity that overlaps with the ABC transporter BcatrD. Bcmfs1 may be involved in protection of B. cinerea against plant defense compounds during the pathogenic phase of growth on host plants and against fungitoxic antimicrobial metabolites during its saprophytic phase of growth. PMID:12324349

  11. Nitric oxide and hydrogen peroxide in tomato resistance. Nitric oxide modulates hydrogen peroxide level in o-hydroxyethylorutin-induced resistance to Botrytis cinerea in tomato.

    PubMed

    Małolepsza, Urszula; Rózalska, Sylwia

    2005-06-01

    Nitric oxide (NO) has been postulated to be required, together with reactive oxygen species (ROS), for activation of disease resistance reactions of plants to infection with a pathogen or elicitor treatment. However, biochemical mechanisms by which ROS and NO participate in these reactions are still under intensive study and controversial debate. We previously demonstrated that o-hydroxyethylorutin when applied on tomato leaves (Lycopersicon esculentum Mill. cv. "Perkoz") restricted Botrytis cinerea infection development. In this research we investigated ROS and NO generation in tomato plants treated with o-hydroxyethylorutin, non-treated and infected ones. The NO content was enhanced or decreased in the studied plants by supplying them with NO generator-SNP or scavenger-cPTIO. NO detection was carried out using diaminofluorescein diacetate (DAF-DA) in conjunction with confocal laser scanning microscopy. The influence of elevated and decreased levels of NO on B. cinerea infection development and ROS generation was studied. The elevated NO concentration in tomato leaves strongly decreased hydrogen peroxide concentration without affecting other studied ROS (superoxide anion and hydroxyl radical) levels. H2O2 concentrations in NO-supplied leaves were low regardless of further treatment of tomato leaves with o-hydroxyethylorutin or inoculation with B. cinerea. The low H2O2 concentration coincided with quick and severe infection development in NO-supplied leaves. As activities of enzymes generating (SOD EC 1.15.1.1)) and removing (APX EC 1.11.1.11, CAT EC 1.11.1.6) H2O2 were unchanged in the studied plants, the decrease in H2O2 concentration was probably due to a direct NO-H2O2 interaction. PMID:15922611

  12. Are Bacterial Volatile Compounds Poisonous Odors to a Fungal Pathogen Botrytis cinerea, Alarm Signals to Arabidopsis Seedlings for Eliciting Induced Resistance, or Both?

    PubMed Central

    Sharifi, Rouhallah; Ryu, Choong-Min

    2016-01-01

    Biological control (biocontrol) agents act on plants via numerous mechanisms, and can be used to protect plants from pathogens. Biocontrol agents can act directly as pathogen antagonists or competitors or indirectly to promote plant induced systemic resistance (ISR). Whether a biocontrol agent acts directly or indirectly depends on the specific strain and the pathosystem type. We reported previously that bacterial volatile organic compounds (VOCs) are determinants for eliciting plant ISR. Emerging data suggest that bacterial VOCs also can directly inhibit fungal and plant growth. The aim of the current study was to differentiate direct and indirect mechanisms of bacterial VOC effects against Botrytis cinerea infection of Arabidopsis. Volatile emissions from Bacillus subtilis GB03 successfully protected Arabidopsis seedlings against B. cinerea. First, we investigated the direct effects of bacterial VOCs on symptom development and different phenological stages of B. cinerea including spore germination, mycelial attachment to the leaf surface, mycelial growth, and sporulation in vitro and in planta. Volatile emissions inhibited hyphal growth in a dose-dependent manner in vitro, and interfered with fungal attachment on the hydrophobic leaf surface. Second, the optimized bacterial concentration that did not directly inhibit fungal growth successfully protected Arabidopsis from fungal infection, which indicates that bacterial VOC-elicited plant ISR has a more important role in biocontrol than direct inhibition of fungal growth on Arabidopsis. We performed qRT-PCR to investigate the priming of the defense-related genes PR1, PDF1.2, and ChiB at 0, 12, 24, and 36 h post-infection and 14 days after the start of plant exposure to bacterial VOCs. The results indicate that bacterial VOCs potentiate expression of PR1 and PDF1.2 but not ChiB, which stimulates SA- and JA-dependent signaling pathways in plant ISR and protects plants against pathogen colonization. This study

  13. Digital Gene Expression Analysis to Screen Disease Resistance-Relevant Genes from Leaves of Herbaceous Peony (Paeonia lactiflora Pall.) Infected by Botrytis cinerea

    PubMed Central

    Gong, Saijie; Hao, Zhaojun; Meng, Jiasong; Liu, Ding; Wei, Mengran; Tao, Jun

    2015-01-01

    Herbaceous peony (Paeonia lactiflora Pall.) is a well-known traditional flower in China and is widely used for landscaping and garden greening due to its high ornamental value. However, disease spots usually appear after the flowering of the plant and may result in the withering of the plant in severe cases. This study examined the disease incidence in an herbaceous peony field in the Yangzhou region, Jiangsu Province. Based on morphological characteristics and molecular data, the disease in this area was identified as a gray mold caused by Botrytis cinerea. Based on previously obtained transcriptome data, eight libraries generated from two herbaceous peony cultivars ‘Zifengyu’ and ‘Dafugui’ with different susceptibilities to the disease were then analyzed using digital gene expression profiling (DGE). Thousands of differentially expressed genes (DEGs) were screened by comparing the eight samples, and these genes were annotated using the Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) database. The pathways related to plant-pathogen interaction, secondary metabolism synthesis and antioxidant system were concentrated, and 51, 76, and 13 disease resistance-relevant candidate genes were identified, respectively. The expression patterns of these candidate genes differed between the two cultivars: their expression of the disease-resistant cultivar ‘Zifengyu’ sharply increased during the early stages of infection, while it was relatively subdued in the disease-sensitive cultivar ‘Dafugui’. A selection of ten candidate genes was evaluated by quantitative real-time PCR (qRT-PCR) to validate the DGE data. These results revealed the transcriptional changes that took place during the interaction of herbaceous peony with B. cinerea, providing insight into the molecular mechanisms of host resistance to gray mold. PMID:26208357

  14. Enhanced Botrytis cinerea Resistance of Arabidopsis Plants Grown in Compost May Be Explained by Increased Expression of Defense-Related Genes, as Revealed by Microarray Analysis

    PubMed Central

    Segarra, Guillem; Santpere, Gabriel; Elena, Georgina; Trillas, Isabel

    2013-01-01

    Composts are the products obtained after the aerobic degradation of different types of organic matter waste and can be used as substrates or substrate/soil amendments for plant cultivation. There is a small but increasing number of reports that suggest that foliar diseases may be reduced when using compost, rather than standard substrates, as growing medium. The purpose of this study was to examine the gene expression alteration produced by the compost to gain knowledge of the mechanisms involved in compost-induced systemic resistance. A compost from olive marc and olive tree leaves was able to induce resistance against Botrytis cinerea in Arabidopsis, unlike the standard substrate, perlite. Microarray analyses revealed that 178 genes were differently expressed, with a fold change cut-off of 1, of which 155 were up-regulated and 23 were down-regulated in compost-grown, as against perlite-grown plants. A functional enrichment study of up-regulated genes revealed that 38 Gene Ontology terms were significantly enriched. Response to stress, biotic stimulus, other organism, bacterium, fungus, chemical and abiotic stimulus, SA and ABA stimulus, oxidative stress, water, temperature and cold were significantly enriched, as were immune and defense responses, systemic acquired resistance, secondary metabolic process and oxireductase activity. Interestingly, PR1 expression, which was equally enhanced by growing the plants in compost and by B. cinerea inoculation, was further boosted in compost-grown pathogen-inoculated plants. Compost triggered a plant response that shares similarities with both systemic acquired resistance and ABA-dependent/independent abiotic stress responses. PMID:23405252

  15. Enhanced Botrytis cinerea resistance of Arabidopsis plants grown in compost may be explained by increased expression of defense-related genes, as revealed by microarray analysis.

    PubMed

    Segarra, Guillem; Santpere, Gabriel; Elena, Georgina; Trillas, Isabel

    2013-01-01

    Composts are the products obtained after the aerobic degradation of different types of organic matter waste and can be used as substrates or substrate/soil amendments for plant cultivation. There is a small but increasing number of reports that suggest that foliar diseases may be reduced when using compost, rather than standard substrates, as growing medium. The purpose of this study was to examine the gene expression alteration produced by the compost to gain knowledge of the mechanisms involved in compost-induced systemic resistance. A compost from olive marc and olive tree leaves was able to induce resistance against Botrytis cinerea in Arabidopsis, unlike the standard substrate, perlite. Microarray analyses revealed that 178 genes were differently expressed, with a fold change cut-off of 1, of which 155 were up-regulated and 23 were down-regulated in compost-grown, as against perlite-grown plants. A functional enrichment study of up-regulated genes revealed that 38 Gene Ontology terms were significantly enriched. Response to stress, biotic stimulus, other organism, bacterium, fungus, chemical and abiotic stimulus, SA and ABA stimulus, oxidative stress, water, temperature and cold were significantly enriched, as were immune and defense responses, systemic acquired resistance, secondary metabolic process and oxireductase activity. Interestingly, PR1 expression, which was equally enhanced by growing the plants in compost and by B. cinerea inoculation, was further boosted in compost-grown pathogen-inoculated plants. Compost triggered a plant response that shares similarities with both systemic acquired resistance and ABA-dependent/independent abiotic stress responses. PMID:23405252

  16. Digital Gene Expression Analysis to Screen Disease Resistance-Relevant Genes from Leaves of Herbaceous Peony (Paeonia lactiflora Pall.) Infected by Botrytis cinerea.

    PubMed

    Gong, Saijie; Hao, Zhaojun; Meng, Jiasong; Liu, Ding; Wei, Mengran; Tao, Jun

    2015-01-01

    Herbaceous peony (Paeonia lactiflora Pall.) is a well-known traditional flower in China and is widely used for landscaping and garden greening due to its high ornamental value. However, disease spots usually appear after the flowering of the plant and may result in the withering of the plant in severe cases. This study examined the disease incidence in an herbaceous peony field in the Yangzhou region, Jiangsu Province. Based on morphological characteristics and molecular data, the disease in this area was identified as a gray mold caused by Botrytis cinerea. Based on previously obtained transcriptome data, eight libraries generated from two herbaceous peony cultivars 'Zifengyu' and 'Dafugui' with different susceptibilities to the disease were then analyzed using digital gene expression profiling (DGE). Thousands of differentially expressed genes (DEGs) were screened by comparing the eight samples, and these genes were annotated using the Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) database. The pathways related to plant-pathogen interaction, secondary metabolism synthesis and antioxidant system were concentrated, and 51, 76, and 13 disease resistance-relevant candidate genes were identified, respectively. The expression patterns of these candidate genes differed between the two cultivars: their expression of the disease-resistant cultivar 'Zifengyu' sharply increased during the early stages of infection, while it was relatively subdued in the disease-sensitive cultivar 'Dafugui'. A selection of ten candidate genes was evaluated by quantitative real-time PCR (qRT-PCR) to validate the DGE data. These results revealed the transcriptional changes that took place during the interaction of herbaceous peony with B. cinerea, providing insight into the molecular mechanisms of host resistance to gray mold. PMID:26208357

  17. Inhibition of SlMPK1, SlMPK2, and SlMPK3 Disrupts Defense Signaling Pathways and Enhances Tomato Fruit Susceptibility to Botrytis cinerea.

    PubMed

    Zheng, Yanyan; Yang, Yang; Liu, Can; Chen, Lin; Sheng, Jiping; Shen, Lin

    2015-06-10

    Mitogen-activated protein kinases (MAPKs) are major components of defense signaling pathways that transduce extracellular stimuli into intracellular responses in plants. Our previous study indicated that SlMPK1/2/3 were associated with nitric oxide-induced defense response in tomato fruit. In this study, we determine whether SlMPK1/2/3 influence the tomato fruit's innate immunity and whether plant hormones and reactive oxygen species (ROS) are involved in SlMPK1/2/3 defense signaling pathways. Treatment with 10 μM U0126 significantly inhibited the relative expression of SlMPK1, SlMPK2, and SlMPK3 (P < 0.05). U0126-treated fruit showed higher concentrations of auxin indole acetic acid (IAA), abscisic acid (ABA), and gibberellic acid (GA), but a lower concentration of methyl jasmonate (MeJA). The activities of defense enzymes, including β-1,3-glucanases (GLU), chitinase (CHI), phenylalanine ammonia lyase (PAL), and polyphenol oxidase (PPO), decreased after U0126 treatment. Meanwhile, H2O2 content increased, and catalase (CAT), ascorbate peroxidase (APX), and peroxidase (POD) activities decreased after U0126 treatment. U0126 treatment enhanced the susceptibility of tomato fruit to Botrytis cinerea and resulted in more severe gray mold rot. These results demonstrate that inhibition of SlMPK1/2/3 disrupts tomato fruit defense signaling pathways and enhances the susceptibility to B. cinerea and also that plant hormones and ROS are associated with SlMPK1/2/3 defense signaling pathways. PMID:25910076

  18. AIR12, a b-type cytochrome of the plasma membrane of Arabidopsis thaliana is a negative regulator of resistance against Botrytis cinerea.

    PubMed

    Costa, Alex; Barbaro, Maria Raffaella; Sicilia, Francesca; Preger, Valeria; Krieger-Liszkay, Anja; Sparla, Francesca; De Lorenzo, Giulia; Trost, Paolo

    2015-04-01

    AIR12 (Auxin Induced in Root culture) is a single gene of Arabidopsis that codes for a mono-heme cytochrome b. Recombinant AIR12 from Arabidopsis accepted electrons from ascorbate or superoxide, and donated electrons to either monodehydroascorbate or oxygen. AIR12 was found associated in vivo to the plasma membrane. Though linked to the membrane by a glycophosphatidylinositol anchor, AIR12 is a hydrophilic and glycosylated protein predicted to be fully exposed to the apoplast. The expression pattern of AIR12 in Arabidopsis is developmentally regulated and correlated to sites of controlled cell separation (e.g. micropilar endosperm during germination, epidermal cells surrounding the emerging lateral root) and cells around wounds. Arabidopsis (Landsberg erecta-0) mutants with altered levels of AIR12 did not show any obvious phenotype. However, AIR12-overexpressing plants accumulated ROS (superoxide, hydrogen peroxide) and lipid peroxides in leaves, indicating that AIR12 may alter the redox state of the apoplast under particular conditions. On the other hand, AIR12-knock out plants displayed a strongly decreased susceptibility to Botrytis cinerea infection, which in turn induced AIR12 expression in susceptible wild type plants. Altogether, the results suggest that AIR12 plays a role in the regulation of the apoplastic redox state and in the response to necrotrophic pathogens. Possible relationships between these functions are discussed. PMID:25711811

  19. Functional analysis of endo-1,4-β-glucanases in response to Botrytis cinerea and Pseudomonas syringae reveals their involvement in plant-pathogen interactions.

    PubMed

    Finiti, I; Leyva, M O; López-Cruz, J; Calderan Rodrigues, B; Vicedo, B; Angulo, C; Bennett, A B; Grant, M; García-Agustín, P; González-Bosch, C

    2013-09-01

    Plant cell wall modification is a critical component in stress responses. Endo-1,4-β-glucanases (EGs) take part in cell wall editing processes, e.g. elongation, ripening and abscission. Here we studied the infection response of Solanum lycopersicum and Arabidopsis thaliana with impaired EGs. Transgenic TomCel1 and TomCel2 tomato antisense plants challenged with Pseudomonas syringae showed higher susceptibility, callose priming and increased jasmonic acid pathway marker gene expression. These two EGs could be resistance factors and may act as negative regulators of callose deposition, probably by interfering with the defence-signalling network. A study of a set of Arabidopsis EG T-DNA insertion mutants challenged with P. syringae and Botrytis cinerea revealed that the lack of other EGs interferes with infection phenotype, callose deposition, expression of signalling pathway marker genes and hormonal balance. We conclude that a lack of EGs could alter plant response to pathogens by modifying the properties of the cell wall and/or interfering with signalling pathways, contributing to generate the appropriate signalling outcomes. Analysis of microarray data demonstrates that EGs are differentially expressed upon many different plant-pathogen challenges, hormone treatments and many abiotic stresses. We found some Arabidopsis EG mutants with increased tolerance to osmotic and salt stress. Our results show that impairing EGs can alter plant-pathogen interactions and may contribute to appropriate signalling outcomes in many different biotic and abiotic plant stress responses. PMID:23528138

  20. Butyl acetate and yeasts interact in adhesion and germination of Botrytis cinerea conidia in vitro and in fungal decay of golden delicious apple.

    PubMed

    Filonow, A B

    2001-04-01

    Butyl acetate is a volatile aroma and flavor compound in apple. Conidia of three strains of Botrytis cinerea, a fungus that causes decay of apple fruit in postharvest storage, had greater adhesion to and greater germination on polycarbonate membrane filters on water inside sealed 500 cc glass jars that were injected with 4 microliters butyl acetate than conidia not so exposed. Conidial germination was highly correlated with conidial adhesion. The yeasts Sporobolomyces roseus and Cryptococcus laurentii, but not Saccharomyces cerevisiae, reduced the adhesion and germination promoting effect of butyl acetate. Conidia did not readily utilize butyl acetate as a food source, as shown by lack of tetrazolium violet reduction, whereas S. roseus and C. laurentii, but not S. cerevisiae did. Butyl acetate added to suspensions of conidia increased the electrical conductivity of the suspensions and increased the loss of 14C from 14C-labeled conidia compared to conidia unexposed to butyl acetate. Uptake of [14C]glucose by conidia was not increased by butyl acetate. Wounds of Golden Delicious apples inoculated with conidia (strain F-J-4) in a dilute solution of butyl acetate had greater decay than unexposed wounds. S. roseus and C. laurentii, but not S. cerevisiae, added with the conidia decreased the incidence or size of decay. Results indicated that butyl acetate increased conidial adhesion, stimulating conidial germination, and some yeasts can reduce this effect. PMID:11446303

  1. BcIqg1, a fungal IQGAP homolog, interacts with NADPH oxidase, MAP kinase and calcium signaling proteins and regulates virulence and development in Botrytis cinerea.

    PubMed

    Marschall, Robert; Tudzynski, Paul

    2016-07-01

    NADPH oxidases (Nox) produce reactive oxygen species (ROS) in multicellular eukaryotic organisms. They trigger defense reactions ('oxidative burst') - in phagocytes and plant cells -, and are involved in a broad range of differentiation processes. Fungal Nox-complexes play a central role in vegetative, sexual and pathogenic processes. In contrast to mammalian systems, knowledge is limited about composition, localisation and connection to major signaling cascades in fungi. Here, we characterize a fungal homolog of the RasGAP scaffold protein IQGAP, which links several major signaling processes, including Nox in mammalian cell lines. We show that BcIqg1 interacts directly with a cytosolic, regulatory component (BcRac) and a membrane-associated subunit (BcNoxD) of a Nox-complex in the pathogen Botrytis cinerea. Thus, this protein may be a scaffold that mediates interaction of the catalytic subunits with the regulator BcNoxR. The protein interacts with modules of the MAP kinase- and calcium-dependent signaling pathways. Functional analysis of BcIqg1 substantiated its involvement in different signaling pathways. It mediates the Ca(2+) -triggered nuclear translocation of - BcCRZ1 and the MAP kinase BcBmp1. BcIqg1 is involved in resistance against oxidative and membrane stress and is required for several developmental processes including formation of sclerotia, conidial anastomosis tubes and infection cushions as well as for virulence. PMID:27062300

  2. [Subcellular localization and resistance to Botrytis cinerea of a new type lipid transfer protein AtDhyPRP1 from Arabidopsis thaliana].

    PubMed

    Zhang, Chen; Li, Lan; Xu, Ziqin

    2012-05-01

    Genetic transformation was adopted to analyze the subcellular localization and the resistance to fungal pathogens of Arabidopsis lipid transfer protein AtDHyPRP1. The coding sequence of AtDHyPRP1 amplified by PCR from Ws ecotype was used to construct the plant binary expression vector pRI101-AN-AtDHyPRP1 and the fusion expression vector pCAMBIA1302-AtDHyPRP1-GFP. Transgenic tobacco and Arabidopsis plants were produced by leaf disc and floral dip protocols, respectively. AtDHyPRP1 could improve the resistance of tobacco to Botrytis cinerea remarkably and the infection sites on transgenic tobacco leaves accumulated large amounts of H2O2. Observation under laser scanning confocal microscope showed that AtDHyPRP1 was localized to cell surface. It suggested that AtDHyPRP1 might play special function after secretion to outside of the cell and was involved in plant defense system against pathogens. PMID:22916498

  3. BcMtg2 is required for multiple stress tolerance, vegetative development and virulence in Botrytis cinerea.

    PubMed

    Shao, Wenyong; Zhang, Yu; Wang, Jin; Lv, Chiyuan; Chen, Changjun

    2016-01-01

    In Saccharomyces cerevisiae, the Mtg2 gene encodes the Obg protein, which has an important function in assembling ribosomal subunits. However, little is known about the role of the Obg GTPase in filamentous fungi. In this study, we identified an Mtg2 ortholog, BcMtg2, in B. cinerea. The BcMtg2 deletion mutant showed a defect in spore production, conidial germination and sclerotial formation. Additionally, the mutant increased sensitivity to various environmental stresses. The BcMtg2 mutant exhibited dramatically decreased virulence on host plant tissues. BcMtg2 mutant showed increased sensitivity to osmotic and oxidative stresses, and to Congo red (cell wall stress agent). In the yeast complement assay, growth defects of yeast BY4741ΔMTG2 mutant were partly restored by genetic complementation of BcMtg2 under these environmental stresses. Additionally, compared with the parental strain and complement strain, the BcMtg2 deletion mutant displayed a minor glycerol response to osmosis stress. These defective phenotypes were recovered in the complement strain ΔBcMtg2C, which was created by adding the wild-type BcMtg2 gene to the ΔBcMtg2 mutant. The results of this study indicate that BcMtg2 has a necessary role in asexual development, environmental stress response and pathogenicity in B. cinerea. PMID:27346661

  4. BcMtg2 is required for multiple stress tolerance, vegetative development and virulence in Botrytis cinerea

    PubMed Central

    Shao, Wenyong; Zhang, Yu; Wang, Jin; Lv, Chiyuan; Chen, Changjun

    2016-01-01

    In Saccharomyces cerevisiae, the Mtg2 gene encodes the Obg protein, which has an important function in assembling ribosomal subunits. However, little is known about the role of the Obg GTPase in filamentous fungi. In this study, we identified an Mtg2 ortholog, BcMtg2, in B. cinerea. The BcMtg2 deletion mutant showed a defect in spore production, conidial germination and sclerotial formation. Additionally, the mutant increased sensitivity to various environmental stresses. The BcMtg2 mutant exhibited dramatically decreased virulence on host plant tissues. BcMtg2 mutant showed increased sensitivity to osmotic and oxidative stresses, and to Congo red (cell wall stress agent). In the yeast complement assay, growth defects of yeast BY4741ΔMTG2 mutant were partly restored by genetic complementation of BcMtg2 under these environmental stresses. Additionally, compared with the parental strain and complement strain, the BcMtg2 deletion mutant displayed a minor glycerol response to osmosis stress. These defective phenotypes were recovered in the complement strain ΔBcMtg2C, which was created by adding the wild-type BcMtg2 gene to the ΔBcMtg2 mutant. The results of this study indicate that BcMtg2 has a necessary role in asexual development, environmental stress response and pathogenicity in B. cinerea. PMID:27346661

  5. Synthesis and in Vitro Antifungal Activity against Botrytis cinerea of Geranylated Phenols and Their Phenyl Acetate Derivatives.

    PubMed

    Chávez, María I; Soto, Mauricio; Taborga, Lautaro; Díaz, Katy; Olea, Andrés F; Bay, Camila; Peña-Cortés, Hugo; Espinoza, Luis

    2015-01-01

    The inhibitory effects on the mycelial growth of plant pathogen Botritys cinerea have been evaluated for a series of geranylphenols substituted with one, two and three methoxy groups in the aromatic ring. The results show that the antifungal activity depends on the structure of the geranylphenols, increasing from 40% to 90% by increasing the number of methoxy groups. On the other hand, the acetylation of the -OH group induces a change of activity that depends on the number of methoxy groups. The biological activity of digeranyl derivatives is lower than that exhibited by the respective monogeranyl compound. All tested geranylphenols have been synthesized by direct coupling of geraniol and the respective phenol. The effect of solvent on yields and product distribution is discussed. For monomethoxyphenols the reaction gives better yields when acetonitrile is used as a solvent and AgNO3 is used as a secondary catalyst. However, for di- and trimethoxyphenols the reaction proceeds only in dioxane. PMID:26287171

  6. Synthesis and in Vitro Antifungal Activity against Botrytis cinerea of Geranylated Phenols and Their Phenyl Acetate Derivatives

    PubMed Central

    Chávez, María I.; Soto, Mauricio; Taborga, Lautaro; Díaz, Katy; Olea, Andrés F.; Bay, Camila; Peña-Cortés, Hugo; Espinoza, Luis

    2015-01-01

    The inhibitory effects on the mycelial growth of plant pathogen Botritys cinerea have been evaluated for a series of geranylphenols substituted with one, two and three methoxy groups in the aromatic ring. The results show that the antifungal activity depends on the structure of the geranylphenols, increasing from 40% to 90% by increasing the number of methoxy groups. On the other hand, the acetylation of the –OH group induces a change of activity that depends on the number of methoxy groups. The biological activity of digeranyl derivatives is lower than that exhibited by the respective monogeranyl compound. All tested geranylphenols have been synthesized by direct coupling of geraniol and the respective phenol. The effect of solvent on yields and product distribution is discussed. For monomethoxyphenols the reaction gives better yields when acetonitrile is used as a solvent and AgNO3 is used as a secondary catalyst. However, for di- and trimethoxyphenols the reaction proceeds only in dioxane. PMID:26287171

  7. Arabidopsis AtERF014 acts as a dual regulator that differentially modulates immunity against Pseudomonas syringae pv. tomato and Botrytis cinerea.

    PubMed

    Zhang, Huijuan; Hong, Yongbo; Huang, Lei; Li, Dayong; Song, Fengming

    2016-01-01

    ERF transcription factors play critical roles in plant immune responses. Here, we report the function of AtERF014, a nucleus-localized transcriptional activator, in Arabidopsis immunity. Expression of AtERF014 was induced by Pseudomonas syringae pv. tomato (Pst) and Botrytis cinerea (Bc). AtERF014-overexpressing (OE) plants displayed increased Pst resistance but decreased Bc resistance, whereas AtERF014-RNAi plants exhibited decreased Pst resistance but increased Bc resistance. After Pst infection, expression of salicylic acid (SA)-responsive genes AtPR1 and AtPR5 in AtERF014-OE plants and of a jasmonic acid/ethylene-responsive gene AtPDF1.2 in AtERF014-RNAi plants was intensified but expression of AtPDF1.2 in AtERF014-OE plants and of AtPR1 and AtPR5 in AtERF014-RNAi plants was weakened. After Bc infection, expression of AtPR1 and AtPR5 in AtERF014-OE plants was attenuated but expression of AtPR1, AtPR5 and AtPDF1.2 in AtERF014-RNAi plants was strengthened. Pathogen- and flg22-induced ROS burst, expression of PTI genes and SA-induced defense were partially suppressed in AtERF014-RNAi plants, whereas pathogen-induced ROS and flg22-induced immune response were strengthened in AtER014-OE plants. Altered expression of AtERR014 affected expression of pectin biosynthetic genes and pectin content in AtERF014-RNAi plants was decreased. These data demonstrate that AtERF014 acts as a dual regulator that differentially modulates immunity against Pst and Bc in Arabidopsis. PMID:27445230

  8. Arabidopsis AtERF014 acts as a dual regulator that differentially modulates immunity against Pseudomonas syringae pv. tomato and Botrytis cinerea

    PubMed Central

    Zhang, Huijuan; Hong, Yongbo; Huang, Lei; Li, Dayong; Song, Fengming

    2016-01-01

    ERF transcription factors play critical roles in plant immune responses. Here, we report the function of AtERF014, a nucleus-localized transcriptional activator, in Arabidopsis immunity. Expression of AtERF014 was induced by Pseudomonas syringae pv. tomato (Pst) and Botrytis cinerea (Bc). AtERF014-overexpressing (OE) plants displayed increased Pst resistance but decreased Bc resistance, whereas AtERF014-RNAi plants exhibited decreased Pst resistance but increased Bc resistance. After Pst infection, expression of salicylic acid (SA)-responsive genes AtPR1 and AtPR5 in AtERF014-OE plants and of a jasmonic acid/ethylene-responsive gene AtPDF1.2 in AtERF014-RNAi plants was intensified but expression of AtPDF1.2 in AtERF014-OE plants and of AtPR1 and AtPR5 in AtERF014-RNAi plants was weakened. After Bc infection, expression of AtPR1 and AtPR5 in AtERF014-OE plants was attenuated but expression of AtPR1, AtPR5 and AtPDF1.2 in AtERF014-RNAi plants was strengthened. Pathogen- and flg22-induced ROS burst, expression of PTI genes and SA-induced defense were partially suppressed in AtERF014-RNAi plants, whereas pathogen-induced ROS and flg22-induced immune response were strengthened in AtER014-OE plants. Altered expression of AtERR014 affected expression of pectin biosynthetic genes and pectin content in AtERF014-RNAi plants was decreased. These data demonstrate that AtERF014 acts as a dual regulator that differentially modulates immunity against Pst and Bc in Arabidopsis. PMID:27445230

  9. Chasing stress signals - Exposure to extracellular stimuli differentially affects the redox state of cell compartments in the wild type and signaling mutants of Botrytis cinerea.

    PubMed

    Marschall, Robert; Schumacher, Julia; Siegmund, Ulrike; Tudzynski, Paul

    2016-05-01

    Reactive oxygen species (ROS) are important molecules influencing intracellular developmental processes as well as plant pathogen interactions. They are produced at the infection site and affect the intracellular redox homeostasis. However, knowledge of ROS signaling pathways, their connection to other signaling cascades, and tools for the visualization of intra- and extracellular ROS levels and their impact on the redox state are scarce. By using the genetically encoded biosensor roGFP2 we studied for the first time the differences between the redox states of the cytosol, the intermembrane space of mitochondria and the ER in the filamentous fungus Botrytis cinerea. We showed that the ratio of oxidized to reduced glutathione inside of the cellular compartments differ and that the addition of hydrogen peroxide (H2O2), calcium chloride (CaCl2) and the fluorescent dye calcofluor white (CFW) have a direct impact on the cellular redox states. Dependent on the type of stress agents applied, the redox states were affected in the different cellular compartments in a temporally shifted manner. By integrating the biosensor in deletion mutants of bcnoxA, bcnoxB, bctrx1 and bcltf1 we further elucidated the putative roles of the different proteins in distinct stress-response pathways. We showed that the redox states of ΔbcnoxA and ΔbcnoxB display a wild-type pattern upon exposure to H2O2, but appear to be strongly affected by CaCl2 and CFW. Moreover, we demonstrated the involvement of the light-responsive transcription factor BcLtf1 in the maintenance of the redox state in the intermembrane space of the mitochondria. Finally, we report that CaCl2 as well as cell wall stress-inducing agents stimulate ROS production and that ΔbcnoxB produces significantly less ROS than the wild type and ΔbcnoxA. PMID:26988904

  10. Resistance to Botrytis cinerea in sitiens, an Abscisic Acid-Deficient Tomato Mutant, Involves Timely Production of Hydrogen Peroxide and Cell Wall Modifications in the Epidermis1[C][W][OA

    PubMed Central

    Asselbergh, Bob; Curvers, Katrien; França, Soraya C.; Audenaert, Kris; Vuylsteke, Marnik; Van Breusegem, Frank; Höfte, Monica

    2007-01-01

    Plant defense mechanisms against necrotrophic pathogens, such as Botrytis cinerea, are considered to be complex and to differ from those that are effective against biotrophs. In the abscisic acid-deficient sitiens tomato (Solanum lycopersicum) mutant, which is highly resistant to B. cinerea, accumulation of hydrogen peroxide (H2O2) was earlier and stronger than in the susceptible wild type at the site of infection. In sitiens, H2O2 accumulation was observed from 4 h postinoculation (hpi), specifically in the leaf epidermal cell walls, where it caused modification by protein cross-linking and incorporation of phenolic compounds. In wild-type tomato plants, H2O2 started to accumulate 24 hpi in the mesophyll layer and was associated with spreading cell death. Transcript-profiling analysis using TOM1 microarrays revealed that defense-related transcript accumulation prior to infection was higher in sitiens than in wild type. Moreover, further elevation of sitiens defense gene expression was stronger than in wild type 8 hpi both in number of genes and in their expression levels and confirmed a role for cell wall modification in the resistant reaction. Although, in general, plant defense-related reactive oxygen species formation facilitates necrotrophic colonization, these data indicate that timely hyperinduction of H2O2-dependent defenses in the epidermal cell wall can effectively block early development of B. cinerea. PMID:17573540

  11. Virus-Induced Gene Silencing-Based Functional Analyses Revealed the Involvement of Several Putative Trehalose-6-Phosphate Synthase/Phosphatase Genes in Disease Resistance against Botrytis cinerea and Pseudomonas syringae pv. tomato DC3000 in Tomato

    PubMed Central

    Zhang, Huijuan; Hong, Yongbo; Huang, Lei; Liu, Shixia; Tian, Limei; Dai, Yi; Cao, Zhongye; Huang, Lihong; Li, Dayong; Song, Fengming

    2016-01-01

    Trehalose and its metabolism have been demonstrated to play important roles in control of plant growth, development, and stress responses. However, direct genetic evidence supporting the functions of trehalose and its metabolism in defense response against pathogens is lacking. In the present study, genome-wide characterization of putative trehalose-related genes identified 11 SlTPSs for trehalose-6-phosphate synthase, 8 SlTPPs for trehalose-6-phosphate phosphatase and one SlTRE1 for trehalase in tomato genome. Nine SlTPSs, 4 SlTPPs, and SlTRE1 were selected for functional analyses to explore their involvement in tomato disease resistance. Some selected SlTPSs, SlTPPs, and SlTRE1 responded with distinct expression induction patterns to Botrytis cinerea and Pseudomonas syringae pv. tomato (Pst) DC3000 as well as to defense signaling hormones (e.g., salicylic acid, jasmonic acid, and a precursor of ethylene). Virus-induced gene silencing-mediated silencing of SlTPS3, SlTPS4, or SlTPS7 led to deregulation of ROS accumulation and attenuated the expression of defense-related genes upon pathogen infection and thus deteriorated the resistance against B. cinerea or Pst DC3000. By contrast, silencing of SlTPS5 or SlTPP2 led to an increased expression of the defense-related genes upon pathogen infection and conferred an increased resistance against Pst DC3000. Silencing of SlTPS3, SlTPS4, SlTPS5, SlTPS7, or SlTPP2 affected trehalose level in tomato plants with or without infection of B. cinerea or Pst DC3000. These results demonstrate that SlTPS3, SlTPS4, SlTPS5, SlTPS7, and SlTPP2 play roles in resistance against B. cinerea and Pst DC3000, implying the importance of trehalose and tis metabolism in regulation of defense response against pathogens in tomato. PMID:27540389

  12. Virus-Induced Gene Silencing-Based Functional Analyses Revealed the Involvement of Several Putative Trehalose-6-Phosphate Synthase/Phosphatase Genes in Disease Resistance against Botrytis cinerea and Pseudomonas syringae pv. tomato DC3000 in Tomato.

    PubMed

    Zhang, Huijuan; Hong, Yongbo; Huang, Lei; Liu, Shixia; Tian, Limei; Dai, Yi; Cao, Zhongye; Huang, Lihong; Li, Dayong; Song, Fengming

    2016-01-01

    Trehalose and its metabolism have been demonstrated to play important roles in control of plant growth, development, and stress responses. However, direct genetic evidence supporting the functions of trehalose and its metabolism in defense response against pathogens is lacking. In the present study, genome-wide characterization of putative trehalose-related genes identified 11 SlTPSs for trehalose-6-phosphate synthase, 8 SlTPPs for trehalose-6-phosphate phosphatase and one SlTRE1 for trehalase in tomato genome. Nine SlTPSs, 4 SlTPPs, and SlTRE1 were selected for functional analyses to explore their involvement in tomato disease resistance. Some selected SlTPSs, SlTPPs, and SlTRE1 responded with distinct expression induction patterns to Botrytis cinerea and Pseudomonas syringae pv. tomato (Pst) DC3000 as well as to defense signaling hormones (e.g., salicylic acid, jasmonic acid, and a precursor of ethylene). Virus-induced gene silencing-mediated silencing of SlTPS3, SlTPS4, or SlTPS7 led to deregulation of ROS accumulation and attenuated the expression of defense-related genes upon pathogen infection and thus deteriorated the resistance against B. cinerea or Pst DC3000. By contrast, silencing of SlTPS5 or SlTPP2 led to an increased expression of the defense-related genes upon pathogen infection and conferred an increased resistance against Pst DC3000. Silencing of SlTPS3, SlTPS4, SlTPS5, SlTPS7, or SlTPP2 affected trehalose level in tomato plants with or without infection of B. cinerea or Pst DC3000. These results demonstrate that SlTPS3, SlTPS4, SlTPS5, SlTPS7, and SlTPP2 play roles in resistance against B. cinerea and Pst DC3000, implying the importance of trehalose and tis metabolism in regulation of defense response against pathogens in tomato. PMID:27540389

  13. Gray Mold or Botrytis Blight

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gray mold, caused by Botrytis cinerea can cause an intermittent decay of leaves or seed production organs of beet. The disease is not usually economically important on sugar beet, but can be severe enough to need control on swiss chard. This chapter describes the disease and pathogen and mentions ...

  14. Overexpression of a Novel NAC Domain-Containing Transcription Factor Gene (AaNAC1) Enhances the Content of Artemisinin and Increases Tolerance to Drought and Botrytis cinerea in Artemisia annua.

    PubMed

    Lv, Zongyou; Wang, Shu; Zhang, Fangyuan; Chen, Lingxian; Hao, Xiaolong; Pan, Qifang; Fu, Xueqing; Li, Ling; Sun, Xiaofen; Tang, Kexuan

    2016-09-01

    The NAC (NAM, ATAF and CUC) superfamily is one of the largest plant-specific transcription factor families. NAC transcription factors always play important roles in response to various abiotic stresses. A NAC transcription factor gene AaNAC1 containing a complete open reading frame (ORF) of 864 bp was cloned from Artemisia annua. The expression of AaNAC1 could be induced by dehydration, cold, salicylic acid (SA) and methyl jasmonate (MJ), suggesting that it might be a key regulator of stress signaling pathways in A. annua. AaNAC1 was shown to be localized to the nuclei by transforming tobacco leaf epidermal cells. When AaNAC1 was overexpressed in A. annua, the content of artemisinin and dihydroartemisinic acid was increased by 79% and 150%, respectively. The expression levels of artemisinin biosynthetic pathway genes, i.e. amorpha-4,11-diene synthase (ADS), artemisinic aldehyde Δ11(13) reductase (DBR2) and aldehyde dehydrogenase 1 (ALDH1), were increased. Dual luciferase (dual-LUC) assays showed that AaNAC1 could activate the transcription of ADS in vivo. The transgenic A. annua exhibited increased tolerance to drought and resistance to Botrytis cinerea. When AaNAC1 was overexpressed in Arabidopsis, the transgenic Arabidopsis were markedly more tolerant to drought. The transgenic Arabidopsis showed increased resistance to B. cinerea. These results indicate that AaNAC1 can potentially be used in transgenic breeding for improving the content of artemisinin and drought tolerance in A. annua. PMID:27388340

  15. Biocontrol Ability and Action Mechanism of Starmerella bacillaris (Synonym Candida zemplinina) Isolated from Wine Musts against Gray Mold Disease Agent Botrytis cinerea on Grape and Their Effects on Alcoholic Fermentation

    PubMed Central

    Lemos, Wilson J.; Bovo, Barbara; Nadai, Chiara; Crosato, Giulia; Carlot, Milena; Favaron, Francesco; Giacomini, Alessio; Corich, Viviana

    2016-01-01

    Gray mold is one of the most important diseases of grapevine in temperate climates. This plant pathogen affects plant growth and reduces wine quality. The use of yeasts as biocontrol agents to apply in the vineyard have been investigated in recent years as an alternative to agrochemicals. In this work, fermenting musts obtained from overripe grape berries, therefore more susceptible to infection by fungal pathogens such as Botrytis cinerea, were considered for the selection of yeasts carrying antifungal activity. Thirty-six isolates were identified as Starmerella bacillaris, a species recently proven to be of enological interest. Among them 14 different strains were studied and antifungal activity against B. cinerea was demonstrated, for the first time, to be present in S. bacillaris species. The production of volatile organic compounds (VOCs), tested in vitro, was found to be the main responsible of S. bacillaris antifungal effects. All the strains were able to reduce B. cinerea decay on wounded grape berries artificially inoculated with gray mold. The colonization level of wound was very high reaching, after 5 days, a concentration of 106 cells per ml of grape juice obtained after berry crushing. At this cell concentration S. bacillaris strains were used to ferment synthetic and natural musts. The sequential yeast inoculation, performed by adding S. cerevisiae 48 h after S. bacillaris, was needed to complete sugar consumption and determined a significant increase in glicerol content and a reduction of ethanol and acetic acid concentrations. The high wound colonization ability, found in this work, together with the propensity to colonize grape berry and the interesting enological traits possessed by the selected S. bacillaris strains allow the use of this yeast as biocontrol agent on vine and grape berries with possible positive effects on must fermentation, although the presence of S. cerevisiae is needed to complete the fermentation process. This work introduces

  16. Fitness and competitive ability of Botrytis cinerea field isolates with dual resistance to SDHI and QoI fungicides, associated with several sdhB and the cytb G143A mutations.

    PubMed

    Veloukas, T; Kalogeropoulou, P; Markoglou, A N; Karaoglanidis, G S

    2014-04-01

    Respiration inhibitors such as the succinate dehydrogenase inhibitors (SDHIs) and the quinone outside inhibitors (QoIs) are fungicide classes with increasing relevance in gray mold control. However, recent studies have shown that dual resistance to both fungicide classes is a common trait in Botrytis cinerea populations from several hosts throughout the world. Resistance of B. cinerea to SDHIs is associated with several mutations in the sdhB, sdhC, and sdhD genes, while resistance to QoIs, in most cases, is associated with the G143A mutation in the cytb gene. The objective of the current study was to investigate the fitness and the competitive ability of B. cinerea field strains possessing one of the H272Y/R/L, N230I, or P225F sdhB substitutions and the G143A mutation of cytb. Fitness parameters measured were (i) mycelial growth and conidia germination in vitro, (ii) aggressiveness and sporulation capacity in vivo, (iii) sclerotia production in vitro and sclerotia viability under different storage conditions, and (iv) sensitivity to oxidative stress imposed by diquat treatments. The competitive ability of the resistant isolates was measured in the absence and presence of the SDHI fungicides boscalid and fluopyram selection pressure. The measurements of individual fitness components showed that the H272R/G143A isolates had the lower differences compared with the sensitive isolates. In contrast, the groups of H272Y/L/G143A, N230I/G143A, and P225F/G143A isolates showed reduced fitness values compared with the sensitive isolates. Isolates possessing only the cytb G143A substitution did not show any fitness cost. The competition experiments showed that, in the absence of fungicide selection pressure, after four disease cycles on apple fruit, the sensitive isolates dominated in the population in all the mixtures tested. In contrast, when the competition experiment was conducted under the selection pressure of boscalid, a gradual decrease in the frequency of sensitive

  17. Biocontrol Ability and Action Mechanism of Starmerella bacillaris (Synonym Candida zemplinina) Isolated from Wine Musts against Gray Mold Disease Agent Botrytis cinerea on Grape and Their Effects on Alcoholic Fermentation.

    PubMed

    Lemos, Wilson J; Bovo, Barbara; Nadai, Chiara; Crosato, Giulia; Carlot, Milena; Favaron, Francesco; Giacomini, Alessio; Corich, Viviana

    2016-01-01

    Gray mold is one of the most important diseases of grapevine in temperate climates. This plant pathogen affects plant growth and reduces wine quality. The use of yeasts as biocontrol agents to apply in the vineyard have been investigated in recent years as an alternative to agrochemicals. In this work, fermenting musts obtained from overripe grape berries, therefore more susceptible to infection by fungal pathogens such as Botrytis cinerea, were considered for the selection of yeasts carrying antifungal activity. Thirty-six isolates were identified as Starmerella bacillaris, a species recently proven to be of enological interest. Among them 14 different strains were studied and antifungal activity against B. cinerea was demonstrated, for the first time, to be present in S. bacillaris species. The production of volatile organic compounds (VOCs), tested in vitro, was found to be the main responsible of S. bacillaris antifungal effects. All the strains were able to reduce B. cinerea decay on wounded grape berries artificially inoculated with gray mold. The colonization level of wound was very high reaching, after 5 days, a concentration of 10(6) cells per ml of grape juice obtained after berry crushing. At this cell concentration S. bacillaris strains were used to ferment synthetic and natural musts. The sequential yeast inoculation, performed by adding S. cerevisiae 48 h after S. bacillaris, was needed to complete sugar consumption and determined a significant increase in glicerol content and a reduction of ethanol and acetic acid concentrations. The high wound colonization ability, found in this work, together with the propensity to colonize grape berry and the interesting enological traits possessed by the selected S. bacillaris strains allow the use of this yeast as biocontrol agent on vine and grape berries with possible positive effects on must fermentation, although the presence of S. cerevisiae is needed to complete the fermentation process. This work

  18. An untargeted global metabolomic analysis reveals the biochemical changes underlying basal resistance and priming in Solanum lycopersicum, and identifies 1-methyltryptophan as a metabolite involved in plant responses to Botrytis cinerea and Pseudomonas syringae.

    PubMed

    Camañes, Gemma; Scalschi, Loredana; Vicedo, Begonya; González-Bosch, Carmen; García-Agustín, Pilar

    2015-10-01

    In this study, we have used untargeted global metabolomic analysis to determine and compare the chemical nature of the metabolites altered during the infection of tomato plants (cv. Ailsa Craig) with Botrytis cinerea (Bot) or Pseudomonas syringae pv. tomato DC3000 (Pst), pathogens that have different invasion mechanisms and lifestyles. We also obtained the metabolome of tomato plants primed using the natural resistance inducer hexanoic acid and then infected with these pathogens. By contrasting the metabolomic profiles of infected, primed, and primed + infected plants, we determined not only the processes or components related directly to plant defense responses, but also inferred the metabolic mechanisms by which pathogen resistance is primed. The data show that basal resistance and hexanoic acid-induced resistance to Bot and Pst are associated with a marked metabolic reprogramming. This includes significant changes in amino acids, sugars and free fatty acids, and in primary and secondary metabolism. Comparison of the metabolic profiles of the infections indicated clear differences, reflecting the fact that the plant's chemical responses are highly adapted to specific attackers. The data also indicate involvement of signaling molecules, including pipecolic and azelaic acids, in response to Pst and, interestingly, to Bot. The compound 1-methyltryptophan was shown to be associated with the tomato-Pst and tomato-Bot interactions as well as with hexanoic acid-induced resistance. Root application of this Trp-derived metabolite also demonstrated its ability to protect tomato plants against both pathogens. PMID:26270176

  19. Elicitin-like proteins Oli-D1 and Oli-D2 from Pythium oligandrum trigger hypersensitive response in Nicotiana benthamiana and induce resistance against Botrytis cinerea in tomato.

    PubMed

    Ouyang, Zhigang; Li, Xiaohui; Huang, Lei; Hong, Yongbo; Zhang, Yafen; Zhang, Huijuan; Li, Dayong; Song, Fengming

    2015-04-01

    The biocontrol agent Pythium oligandrum and its elicitin-like proteins oligandrins have been shown to induce disease resistance in a range of plants. In the present study, the ability of two oligandrins, Oli-D1 and Oli-D2, to induce an immune response and the possible molecular mechanism regulating the defence responses in Nicotiana benthamiana and tomato were investigated. Infiltration of recombinant Oli-D1 and Oli-D2 proteins induced a typical immune response in N. benthamiana including the induction of a hypersensitive response (HR), accumulation of reactive oxygen species and production of autofluorescence. Agrobacterium-mediated transient expression assays revealed that full-length Oli-D1 and Oli-D2 were required for full HR-inducing activity in N. benthamiana, and virus-induced gene silencing-mediated knockdown of some of the signalling regulatory genes demonstrated that NbSGT1 and NbNPR1 were required for Oli-D1 and Oli-D2 to induce HR in N. benthamiana. Subcellular localization analyses indicated that both Oli-D1 and Oli-D2 were targeted to the plasma membrane of N. benthamiana. When infiltrated or transiently expressed in leaves, Oli-D1 and Oli-D2 induced resistance against Botrytis cinerea in tomato and activated the expression of a set of genes involved in the jasmonic acid/ethylene (JA/ET)-mediated signalling pathway. Our results demonstrate that Oli-D1 and Oli-D2 are effective elicitors capable of inducing immune responses in plants, probably through the JA/ET-mediated signalling pathway, and that both Oli-D1 and Oli-D2 have potential for the development of bioactive formulae for crop disease control in practice. PMID:25047132

  20. Efficiacy of bumble bee disseminated biological control agents for control of Botrytis Blossom blight of Rabbiteye Blueberry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis blossom blight caused by Botrytis cinerea may cause severe crop loss in rabbiteye blueberry, necessitating applications of expensive fungicides. Commercial bumble bees, Bombus impatiens, were tested as vectors of the fungicidal biological control agents (BCAs), Prestop® Gliocladium catenula...

  1. Development of PCR-based assays for detecting and differentiating three species of botrytis infecting broad bean

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis cinerea, B. fabae and B. fabiopsis are known to cause chocolate spot on broad bean. This study was conducted to develop PCR-based assays to detect and differentiate this three species. Two sets of primers, Bc-f/Bc-r for B. cinerea and Bfab-f/Bfab-r for B. fabiopsis, were designed based on t...

  2. Analysis of Cryptic, Systemic Botrytis Infections in Symptomless Hosts

    PubMed Central

    Shaw, Michael W.; Emmanuel, Christy J.; Emilda, Deni; Terhem, Razak B.; Shafia, Aminath; Tsamaidi, Dimitra; Emblow, Mark; van Kan, Jan A. L.

    2016-01-01

    Botrytis species are generally considered to be aggressive, necrotrophic plant pathogens. By contrast to this general perception, however, Botrytis species could frequently be isolated from the interior of multiple tissues in apparently healthy hosts of many species. Infection frequencies reached 50% of samples or more, but were commonly less, and cryptic infections were rare or absent in some plant species. Prevalence varied substantially from year to year and from tissue to tissue, but some host species routinely had high prevalence. The same genotype was found to occur throughout a host, representing mycelial spread. Botrytis cinerea and Botrytis pseudocinerea are the species that most commonly occur as cryptic infections, but phylogenetically distant isolates of Botrytis were also detected, one of which does not correspond to previously described species. Sporulation and visible damage occurred only when infected tissues were stressed, or became mature or senescent. There was no evidence of cryptic infection having a deleterious effect on growth of the host, and prevalence was probably greater in plants grown in high light conditions. Isolates from cryptic infections were often capable of causing disease (to varying extents) when spore suspensions were inoculated onto their own host as well as on distinct host species, arguing against co-adaptation between cryptic isolates and their hosts. These data collectively suggest that several Botrytis species, including the most notorious pathogenic species, exist frequently in cryptic form to an extent that has thus far largely been neglected, and do not need to cause disease on healthy hosts in order to complete their life-cycles. PMID:27242829

  3. Antagonistic Effect of Streptomyces sp. BS062 against Botrytis Diseases

    PubMed Central

    Kim, Young-Sook; Lee, In-Kyoung

    2015-01-01

    The use of microorganisms and their secreted molecules to prevent plant diseases is considered an attractive alternative and way to supplement synthetic fungicides for the management of plant diseases. Strain BS062 was selected based on its ability to inhibit the mycelial growth of Botrytis cinerea, a major causal fungus of postharvest root rot of ginseng and strawberry gray mold disease. Strain BS062 was found to be closely related to Streptomyces hygroscopicus (99% similarity) on the basis of 16S ribosomal DNA sequence analysis. Postharvest root rot of ginseng and strawberry gray mold disease caused by B. cinerea were controlled up to 73.9% and 58%, respectively, upon treatment with culture broth of Streptomyces sp. BS062. These results suggest that strain BS062 may be a potential agent for controlling ginseng postharvest root rot and strawberry gray mold disease. PMID:26539052

  4. Phenotypical differences among B. cinerea isolates from ornamental plants.

    PubMed

    Martínez, J A; Valdés, R; Vicente, M J; Bañón, S

    2008-01-01

    B. cinerea is a common pathogenic fungus which causes Botrytis blight (Grey mould) in most ornamental plants. It may be responsible for serious preharvest diseases and postharvest losses in fruits, vegetables and flowers. In this work, several B. cinerea isolates from ornamental plants (Chamelaucium uncinatum, Pelargonium x hortorum, Euphorbia pulcherrima, Lantana camara, Lonicera japonica, Hydrangea macrophylla, and Cyclamen persicum) affected by Botrytis blight in the south of Spain were studied. All the isolates were confirmed as B. cinerea by PCR using a specific 750-bp molecular marker, which is present in all strains of this species but absent from other species of Botrytis. The isolates were evaluated by reference to mature conidia length, sclerotia production, and growth rate. Conidia, conidiophores and hyphae were described by light microscopy and some by cryogenic scanning electron microscopy (Cryo-SEM). Conidium length was measured by using an eyepiece micrometer at 400x power, whereas the growth rate was assessed from differences in colony diameter between the third and fourth day of growth in potato-dextrose agar culture medium at 26 degrees C. B. cinerea showed a high degree of phenotypical variability among isolates, not only as regards visual aspects of the colonies but also in some morphological structures such as conidium length, conidiophores, sclerotia production, and hyphae. Differences were also observed in the growth rates. Conidiation was insignificant in the isolates from H. macrophylla, and P. x hortorum, where the overall appearance was white in all the growing stages, whereas isolates from L. camara, C. persicum and C. uncinatum were mainly grey or brown in mature stages. The longest conidia were obtained in isolates from H. macrophylla and C. persicum (17-18 microm) and the lowest in C. uncinatum (9 microm). All the isolates, except L. camara, developed mature sclerotia after approximately 16 days in the conditions used. H. macrophylla

  5. Inflorescences vs leaves: a distinct modulation of carbon metabolism process during Botrytis infection.

    PubMed

    Vatsa-Portugal, Parul; Walker, Anne-Sophie; Jacquens, Lucile; Clément, Christophe; Barka, Essaid Ait; Vaillant-Gaveau, Nathalie

    2015-05-01

    Plant growth and survival depends critically on photo assimilates. Pathogen infection leads to changes in carbohydrate metabolism of plants. In this study, we monitored changes in the carbohydrate metabolism in the grapevine inflorescence and leaves using Botrytis cinerea and Botrytis pseudo cinerea. Fluctuations in gas exchange were correlated with variations in chlorophyll a fluorescence. During infection, the inflorescences showed an increase in net photosynthesis (Pn) with a stomatal limitation. In leaves, photosynthesis decreased, with a non-stomatal limitation. A decrease in the effective photosystem II (PSII) quantum yield (ΦPSII) was accompanied by an increase in photochemical quenching (qP) and non-photochemical quenching (qN). The enhancement of qP and ΦPSII could explain the observed increase in Pn. In leaves, the significant decline in ΦPSII and qP, and increase in qN suggest that energy was mostly oriented toward heat dissipation instead of CO2 fixation. The accumulation of glucose and sucrose in inflorescences and glucose and fructose in the leaves during infection indicate that the plant's carbon metabolism is differently regulated in these two organs. While a strong accumulation of starch was observed at 24 and 48 hours post-inoculation (hpi) with both species of Botrytis in the inflorescences, a significant decrease with B. cinerea at 24 hpi and a significant increase with B. pseudo cinerea at 48 hpi were observed in the leaves. On the basis of these results, it can be said that during pathogen attack, the metabolism of grapevine inflorescence and leaf is modified suggesting distinct mechanisms modifying gas exchange, PSII activity and sugar contents in these two organs. PMID:25251162

  6. The rising threat of fungicide resistance in plant pathogenic fungi: Botrytis as a case study.

    PubMed

    Hahn, Matthias

    2014-10-01

    The introduction of site-specific fungicides almost 50 years ago has revolutionized chemical plant protection, providing highly efficient, low toxicity compounds for control of fungal diseases. However, it was soon discovered that plant pathogenic fungi can adapt to fungicide treatments by mutations leading to resistance and loss of fungicide efficacy. The grey mould fungus Botrytis cinerea, a major cause of pre- and post-harvest losses in fruit and vegetable production, is notorious as a 'high risk' organism for rapid resistance development. In this review, the mechanisms and the history of fungicide resistance in Botrytis are outlined. The introduction of new fungicide classes for grey mould control was always followed by the appearance of resistance in field populations. In addition to target site resistance, B. cinerea has also developed a resistance mechanism based on drug efflux transport. Excessive spraying programmes have resulted in the selection of multiresistant strains in several countries, in particular in strawberry fields. The rapid erosion of fungicide activity against these strains represents a major challenge for the future of fungicides against Botrytis. To maintain adequate protection of intensive cultures against grey mould, strict implementation of resistance management measures are required as well as alternative strategies with non-chemical products. PMID:25320647

  7. Fungal endopolygalacturonases are recognized as microbe-associated molecular patterns by the arabidopsis receptor-like protein RESPONSIVENESS TO BOTRYTIS POLYGALACTURONASES1.

    PubMed

    Zhang, Lisha; Kars, Ilona; Essenstam, Bert; Liebrand, Thomas W H; Wagemakers, Lia; Elberse, Joyce; Tagkalaki, Panagiota; Tjoitang, Devlin; van den Ackerveken, Guido; van Kan, Jan A L

    2014-01-01

    Plants perceive microbial invaders using pattern recognition receptors that recognize microbe-associated molecular patterns. In this study, we identified RESPONSIVENESS TO BOTRYTIS POLYGALACTURONASES1 (RBPG1), an Arabidopsis (Arabidopsis thaliana) leucine-rich repeat receptor-like protein, AtRLP42, that recognizes fungal endopolygalacturonases (PGs) and acts as a novel microbe-associated molecular pattern receptor. RBPG1 recognizes several PGs from the plant pathogen Botrytis cinerea as well as one from the saprotroph Aspergillus niger. Infiltration of B. cinerea PGs into Arabidopsis accession Columbia induced a necrotic response, whereas accession Brno (Br-0) showed no symptoms. A map-based cloning strategy, combined with comparative and functional genomics, led to the identification of the Columbia RBPG1 gene and showed that this gene is essential for the responsiveness of Arabidopsis to the PGs. Transformation of RBPG1 into accession Br-0 resulted in a gain of PG responsiveness. Transgenic Br-0 plants expressing RBPG1 were equally susceptible as the recipient Br-0 to the necrotroph B. cinerea and to the biotroph Hyaloperonospora arabidopsidis. Pretreating leaves of the transgenic plants with a PG resulted in increased resistance to H. arabidopsidis. Coimmunoprecipitation experiments demonstrated that RBPG1 and PG form a complex in Nicotiana benthamiana, which also involves the Arabidopsis leucine-rich repeat receptor-like protein SOBIR1 (for SUPPRESSOR OF BIR1). sobir1 mutant plants did not induce necrosis in response to PGs and were compromised in PG-induced resistance to H. arabidopsidis. PMID:24259685

  8. Fungal Endopolygalacturonases Are Recognized as Microbe-Associated Molecular Patterns by the Arabidopsis Receptor-Like Protein RESPONSIVENESS TO BOTRYTIS POLYGALACTURONASES11[W

    PubMed Central

    Zhang, Lisha; Kars, Ilona; Essenstam, Bert; Liebrand, Thomas W.H.; Wagemakers, Lia; Elberse, Joyce; Tagkalaki, Panagiota; Tjoitang, Devlin; van den Ackerveken, Guido; van Kan, Jan A.L.

    2014-01-01

    Plants perceive microbial invaders using pattern recognition receptors that recognize microbe-associated molecular patterns. In this study, we identified RESPONSIVENESS TO BOTRYTIS POLYGALACTURONASES1 (RBPG1), an Arabidopsis (Arabidopsis thaliana) leucine-rich repeat receptor-like protein, AtRLP42, that recognizes fungal endopolygalacturonases (PGs) and acts as a novel microbe-associated molecular pattern receptor. RBPG1 recognizes several PGs from the plant pathogen Botrytis cinerea as well as one from the saprotroph Aspergillus niger. Infiltration of B. cinerea PGs into Arabidopsis accession Columbia induced a necrotic response, whereas accession Brno (Br-0) showed no symptoms. A map-based cloning strategy, combined with comparative and functional genomics, led to the identification of the Columbia RBPG1 gene and showed that this gene is essential for the responsiveness of Arabidopsis to the PGs. Transformation of RBPG1 into accession Br-0 resulted in a gain of PG responsiveness. Transgenic Br-0 plants expressing RBPG1 were equally susceptible as the recipient Br-0 to the necrotroph B. cinerea and to the biotroph Hyaloperonospora arabidopsidis. Pretreating leaves of the transgenic plants with a PG resulted in increased resistance to H. arabidopsidis. Coimmunoprecipitation experiments demonstrated that RBPG1 and PG form a complex in Nicotiana benthamiana, which also involves the Arabidopsis leucine-rich repeat receptor-like protein SOBIR1 (for SUPPRESSOR OF BIR1). sobir1 mutant plants did not induce necrosis in response to PGs and were compromised in PG-induced resistance to H. arabidopsidis. PMID:24259685

  9. Calmodulin gene expression in response to mechanical wounding and Botrytis cinerea infection in tomato fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Calmodulin, a ubiquitous calcium sensor, plays an important role in decoding the stress-triggered intracellular calcium changes and regulates the functions of numerous target proteins involved in various physiological responses in plants. To determine the functional significance of calmodulin in fl...

  10. Polyamines attenuate ethylene-mediated defense responses to abrogate resistance to Botrytis cinerea in tomato

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Transgenic tomato (Solanum lycopersicum) lines over-expressing yeast spermidine synthase (ySpdSyn), an enzyme involved in polyamine (PA) biosynthesis, were developed. These transgenic lines accumulate higher levels of spermidine (Spd) than the wild type plants and were examined for responses to the...

  11. Botrytis pyriformis sp. nov., a novel and likely saprophytic species of Botrytis.

    PubMed

    Zhang, J; Yang, H; Yu, Q Y; Wu, M D; Yang, L; Zhuang, W Y; Chen, W D; Li, G Q

    2016-01-01

    A novel species of Botrytis from Sedum sarmentosum was described based on morphology and analyses of DNA sequences of nuc rDNA ITS regions and three nuclear genes (G3PDH, HSP60, RPB2). Meanwhile pathogenicity in 32 plant species, response to temperature for growth and conidial germination for the species were determined. The Botrytis species was named Botrytis pyriformis sp. nov. It was characterized by formation of grayish mycelia, brownish conidia and melanized sclerotia on PDA. The conidia are pear-shaped, melanized and covered with abundant villiform appendages on the conidial surface. Comparison of the ITS sequences confirmed its placement in the genus Botrytis Phylogenetic analysis based on DNA sequences of G3PDH, HSP60 and RPB2 genes indicated that B. pyriformis and other 30 Botrytis species form a monophyletic clade, which was further divided into three subclades. Subclade I comprised B. pyriformis alone, whereas subclades II and III comprised six and 24 Botrytis species, respectively. Botrytis pyriformis could not infect 32 plant species including S. sarmentosum, possibly due to deficiency in formation of infection cushions. This study presents a formal description and illustrations for B. pyriformis and provides experimental evidence, indicating that B. pyriformis might be a saprophytic species. PMID:27153884

  12. Cryopreservation of Juglans cinerea (butternut) dormant buds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Juglans cinerea (butternut) is a deciduous tree native to the United States and Canada with oblong-lemon shaped nuts with oily texture and pleasant flavor. Butternut wood is softer than wood of the black walnut making it a favorite wood for woodcarvers. In North America butternut is seriously thre...

  13. The Arabidopsis Botrytis Susceptible1 Interactor Defines a Subclass of RING E3 Ligases That Regulate Pathogen and Stress Responses1[C][W

    PubMed Central

    Luo, Hongli; Laluk, Kristin; Lai, Zhibing; Veronese, Paola; Song, Fengming; Mengiste, Tesfaye

    2010-01-01

    We studied the function of Arabidopsis (Arabidopsis thaliana) Botrytis Susceptible1 Interactor (BOI) in plant responses to pathogen infection and abiotic stress. BOI physically interacts with and ubiquitinates Arabidopsis BOS1, an R2R3MYB transcription factor previously implicated in stress and pathogen responses. In transgenic plants expressing the BOS1-β-glucuronidase transgene, β-glucuronidase activity could be detected only after inhibition of the proteosome, suggesting that BOS1 is a target of ubiquitin-mediated degradation by the proteosome. Plants with reduced BOI transcript levels generated through RNA interference (BOI RNAi) were more susceptible to the necrotrophic fungus Botrytis cinerea and less tolerant to salt stress. In addition, BOI RNAi plants exhibited increased cell death induced by the phytotoxin α-picolinic acid and by a virulent strain of the bacterial pathogen Pseudomonas syringae, coincident with peak disease symptoms. However, the hypersensitive cell death associated with different race-specific resistance genes was unaffected by changes in the level of BOI transcript. BOI expression was enhanced by B. cinerea and salt stress but repressed by the plant hormone gibberellin, indicating a complex regulation of BOI gene expression. Interestingly, BOI RNAi plants exhibit reduced growth responsiveness to gibberellin. We also present data revealing the function of three Arabidopsis BOI-RELATED GENES (BRGs), which contribute to B. cinerea resistance and the suppression of disease-associated cell death. In sum, BOI and BRGs represent a subclass of RING E3 ligases that contribute to plant disease resistance and abiotic stress tolerance through the suppression of pathogen-induced as well as stress-induced cell death. PMID:20921156

  14. Efficacy of Combined Formulations of Fungicides with Different Modes of Action in Controlling Botrytis Gray Mold Disease in Chickpea

    PubMed Central

    Rashid, M. H.; Hossain, M. Ashraf; Kashem, M. A.; Kumar, Shiv; Rafii, M. Y.; Latif, M. A.

    2014-01-01

    Botrytis gray mold (BGM) caused by Botrytis cinerea Pers. Ex. Fr. is an extremely devastating disease of chickpea (Cicer arietinum L.) and has a regional as well as an international perspective. Unfortunately, nonchemical methods for its control are weak and ineffective. In order to identify an effective control measure, six fungicides with different modes of action were evaluated on a BGM susceptible chickpea variety BARIchhola-1 at a high BGM incidence location (Madaripur) in Bangladesh for three years (2008, 2009, and 2010). Among the six fungicides tested, one was protectant [Vondozeb 42SC, a.i. mancozeb (0.2%)], two systemic [Bavistin 50 WP, a.i. carbendazim (0.2%), and Protaf 250EC, propiconazole (0.05%)], and three combination formulations [Acrobat MZ690, dimethomorph 9% + mancozeb 60%, (0.2%); Secure 600 WG, phenomadone + mancozeb (0.2%); and Companion, mancozeb 63% + carbendazim 12% (0.2%)]. The results showed superiority of combination formulations involving both protectant and systemic fungicides over the sole application of either fungicide separately. Among the combination fungicides, Companion was most effective, resulting in the lowest disease severity (3.33 score on 1–9 scale) and the highest increase (38%) of grain yield in chickpea. Therefore, this product could be preferred over the sole application of either solo protectant or systemic fungicides to reduce yield losses and avoid fungicide resistance. PMID:24723819

  15. The Membrane-Anchored BOTRYTIS-INDUCED KINASE1 Plays Distinct Roles in Arabidopsis Resistance to Necrotrophic and Biotrophic PathogensW⃞

    PubMed Central

    Veronese, Paola; Nakagami, Hirofumi; Bluhm, Burton; AbuQamar, Synan; Chen, Xi; Salmeron, John; Dietrich, Robert A.; Hirt, Heribert; Mengiste, Tesfaye

    2006-01-01

    Plant resistance to disease is controlled by the combination of defense response pathways that are activated depending on the nature of the pathogen. We identified the Arabidopsis thaliana BOTRYTIS-INDUCED KINASE1 (BIK1) gene that is transcriptionally regulated by Botrytis cinerea infection. Inactivation of BIK1 causes severe susceptibility to necrotrophic fungal pathogens but enhances resistance to a virulent strain of the bacterial pathogen Pseudomonas syringae pv tomato. The response to an avirulent bacterial strain is unchanged, limiting the role of BIK1 to basal defense rather than race-specific resistance. The jasmonate- and ethylene-regulated defense response, generally associated with resistance to necrotrophic fungi, is attenuated in the bik1 mutant based on the expression of the plant defensin PDF1.2 gene. bik1 mutants show altered root growth, producing more and longer root hairs, demonstrating that BIK1 is also required for normal plant growth and development. Whereas the pathogen responses of bik1 are mostly dependent on salicylic acid (SA) levels, the nondefense responses are independent of SA. BIK1 is membrane-localized, suggesting possible involvement in early stages of the recognition or transduction of pathogen response. Our data suggest that BIK1 modulates the signaling of cellular factors required for defense responses to pathogen infection and normal root hair growth, linking defense response regulation with that of growth and development. PMID:16339855

  16. The complete mitochondrial genome of Motacilla cinerea (Passeriformes: Motacillidae).

    PubMed

    Zhang, Zhen; Qian, Lifu; Wang, Yupeng; Zhang, Baowei

    2016-09-01

    Motacilla cinerea is a species of small- and medium-sized songbird in the Family Motacillidae, which is widely distributed. In this study, we determined the complete mitochondrial genome of M. cinerea. The result showed that the total length of the mitogenome was 16 825 bp and contained two ribosomal RNA genes, 22 transfer RNA genes, 13 protein-coding genes, and one control region. All the genes in M. cinerea were distributed on the H-strand, except for the ND6 subunit gene and eight tRNA genes which are encoded on the L-strand. The phylogenetic tree was reconstructed using Bayesian analysis methods, and containing two clades: Motacilla and Anthus. The first lineage is Motacilla including M. cinerea and other nine species. The genus Anthus makes up the second group, which containing 17 species. PMID:26328907

  17. Regulation of Botrytis cinerea virulence genes in interaction with Trichoderma arundinaceum is mediated by the sesquiterpene harzianum A

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Trichoderma includes a great diversity of species, some of them with the ability to control the growth of fungal phytopathogens. Many of these strains produce secondary metabolites that are able to inhibit the growth of their fungal preys. However, pathogens can also produce metabolites that in some...

  18. A new and reliable method for live imaging and quantification of reactive oxygen species in Botrytis cinerea: technological advancement.

    PubMed

    Marschall, Robert; Tudzynski, Paul

    2014-10-01

    Reactive oxygen species (ROS) are produced in conserved cellular processes either as by-products of the cellular respiration in mitochondria, or purposefully for defense mechanisms, signaling cascades or cell homeostasis. ROS have two diametrically opposed attributes due to their highly damaging potential for DNA, lipids and other molecules and due to their indispensability for signaling and developmental processes. In filamentous fungi, the role of ROS in growth and development has been studied in detail, but these analyses were often hampered by the lack of reliable and specific techniques to monitor different activities of ROS in living cells. Here, we present a new method for live cell imaging of ROS in filamentous fungi. We demonstrate that by use of a mixture of two fluorescent dyes it is possible to monitor H2O2 and superoxide specifically and simultaneously in distinct cellular structures during various hyphal differentiation processes. In addition, the method allows for reliable fluorometric quantification of ROS. We demonstrate that this can be used to characterize different mutants with respect to their ROS production/scavenging potential. PMID:25220147

  19. Polyamine spermidine is an upstream negator of ethylene-regulated pathogenesis of botrytis cinerea in tomato leaf

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polyamines are biogenic polycationic compounds implicated in a number of processes including plant cell division, cell elongation, flowering, fruit set and development, fruit ripening, senescence and responses to abiotic stresses. Comparatively, little is known about their role in plant-microbe int...

  20. Novel aspinolide production by Trichoderma arundinaceum with a potential role in Botrytis cinerea antagonistic activity and plant defense priming

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Harzianum A (HA), a trichothecene produced by Trichoderma arundinaceum, has recently been described to have antagonistic activity against fungal plant pathogens and to induce plant defence genes. In the present work, we have shown that a tri5 genedisrupted mutant that lacks HA production overproduce...

  1. HPLC analysis and cytotoxic activity of Vernonia cinerea.

    PubMed

    Khay, Mom; Toeng, Phirom; Mahiou-Leddet, Valérie; Mabrouki, Fathi; Sothea, Kim; Ollivier, Evelyne; Elias, Riad; Bun, Sok-Siya

    2012-10-01

    The extracts of five Cambodian medicinal plants (Aganosma marginata, Dracaena cambodiana, Harrisonia perforata, Hymenodictyon excelsum and Vernonia cinerea) were evaluated in vitro for their cytotoxic activity against HT29 colon adenocarcinoma cells and HepG2 hepatoma cells, using the MTT assay. Among these five plants, Vernonia cinerea displayed potent cytotoxicity. One main sesquiterpene lactone, 8alpha-tigloyloxy-hirsutinolide-13-O-acetate was isolated from the whole plant of V. cinerea. This compound was active against both cancer cell lines (IC50 = 3.50 microM for HT29 and IC50 = 4.27 microM for HepG2). To quantify this compound in the plant, an analytical high-performance liquid chromatography (HPLC) method was developed and validated. PMID:23156983

  2. Suppression of Botrytis Gray Mold in Hiemalis Begonia with Potting Mix Amendments

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis gray mold, arguably the most prevalent disease of hiemalis begonia in the greenhouse, causes lesions on flowers, leaves and stems throughout the cropping cycle. Traditional control recommendations rely heavily on cultural practices and the regular application of fungicides. Since Botrytis...

  3. Meningitis and Bacteremia Due to Neisseria cinerea following a Percutaneous Rhizotomy of the Trigeminal Ganglion

    PubMed Central

    Richter, H.; Bruderer, T.; Goldenberger, D.; Emonet, S.; Strahm, C.

    2015-01-01

    Neisseria cinerea is a human commensal. The first known case of meningitis and bacteremia due to Neisseria cinerea following percutaneous glycerol instillation of the trigeminal ganglion is reported. Conventional phenotypic methods and complete 16S RNA gene sequencing accurately identified the pathogen. Difficulties in differentiation from pathogenic neisseriae are discussed. PMID:26511743

  4. Fatal bacteremia by neisseria cinerea in a woman with myelodysplastic syndrome: a case report

    PubMed Central

    Zhu, Xiaofei; Li, Min; Cao, Huiling; Yang, Xuewen

    2015-01-01

    Neisseria cinerea has been rarely found in blood cultures. In this study, we are reporting a case of a Myelodysplastic Syndrome (MDS) patient in whose blood Neisseria cinerea was found and led a fatal consequence. This case will call our attentions to the uncommon pathogens in the pathogenicity of end-stage patients. PMID:26131259

  5. Multiple endosymbionts in populations of the ant Formica cinerea

    PubMed Central

    2010-01-01

    Background Many insects, including ants, are infected by maternally inherited Wolbachia endosymbiotic bacteria though other secondary endosymbionts have not been reported in ants. It has been suggested that the ability of Wolbachia to invade and remain in an ant population depends on the number of coexisting queens in a colony. We study the genetic and social structure of populations in the ant Formica cinerea which is known to have populations with either monogynous or polygynous colonies. We screen populations for several endosymbiotic bacteria to evaluate the presence of different endosymbionts, possible association between their prevalence and the social structure, and the association between endosymbiont prevalence and genetic differentiation of ant populations. Results We found three endosymbiotic bacteria; 19% of the nests were infected by Wolbachia, 3.8% by Cardinium and 33% by Serratia. There was significant variation among the populations regarding the proportion of nests infected by Serratia, Wolbachia and the pooled set of all the endosymbionts. Some individuals and colonies carried two of the bacteria, the frequency of double infections agreeing with the random expectation. The proportion of infected ants (individuals or colonies) did not correlate significantly with the population level relatedness values. The difference in the prevalence of Wolbachia between population pairs correlated significantly with the genetic distance (microsatellites) of the populations. Conclusions The discovery of several endosymbionts and co-infections by Wolbachia and Cardinium demonstrate the importance of screening several endosymbionts when evaluating their possible effects on social life and queen-worker conflicts over sex allocation. The low prevalence of Wolbachia in F. cinerea departs from the pattern observed in many other Formica ants in which all workers have been infected. It is likely that the strain of Wolbachia in F. cinerea differs from those in other

  6. Thirteen nuclear microsatellite loci for butternut (Juglans cinerea L.).

    PubMed

    Hoban, Sean; Anderson, Robert; McCleary, Tim; Schlarbaum, Scott; Romero-Severson, Jeanne

    2008-05-01

    Butternut (Juglans cinerea L.) is an eastern North American forest tree severely threatened by an exotic fungal pathogen, Sirococcus clavigignenti-juglandacearum. We report here 13 nuclear microsatellites for genetic evaluation of the remaining natural populations. Summary statistics are reported for individuals from a population of butternuts in central Kentucky (N = 63). All markers were polymorphic, with an average of 13.7 alleles per locus observed. Four loci exhibited significantly fewer heterozygotes than expected under Hardy-Weinberg equilibrium (P < 0.05). PMID:21585858

  7. A review on ethno-medicinal uses and pharmacology of Vernonia cinerea Less.

    PubMed

    Dogra, Nittya K; Kumar, Suresh

    2015-01-01

    Vernonia cinerea Less. (ash-coloured fleabane; Asteraceae) is a widely distributed plant throughout India. The plant has reputation as folklore medicine in various traditional systems of medicine. The plant has been evaluated for varied pharmacological activities to validate its traditional claims, and has been scientifically reported to possess anti-inflammatory, antidiabetic, renoprotective, anticancer, antiviral, antimicrobial activities, etc. This review emphasises on ethnopharmacology and pharmacology of V. cinerea. PMID:25426778

  8. The potential role of PR-8 gene of apple fruit in the mode of action of yeast antagonist, Candida oleophila, in postharvest biocontrol of Botrytis cinerea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several pathogenesis-related (PR) genes of apple have been cloned and identified in response to specific pathogens. However, different PR genes in certain organs of specific host may be involved in specific interaction with different microbes. Current research is aimed at characterizing specific P...

  9. Impact of Applied Nitrogen Concentration on Growth of Elatior Begonia and New Guinea Impatiens, and Susceptibility of Begonia to Botrytis cinerea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant performance and appearance in deficient and toxic levels of nutrients are well characterized. However, less is known about the potential subtleties of plant growth, form, development, nutrient uptake, and biotic stress tolerance in the broad tolerable range. Begonia (Beg, Begonia × tuberhybr...

  10. Laboratory evaluation of molluscicidal activity of extracts from Cotula cinerea (L) and Quercus lusitania var. infectoria galls (Oliv.).

    PubMed

    Redwane, A; Markouk, M; Lazrek, H B; Amarouch, H; Jana, M

    1998-01-01

    In this work, we have studied the molluscicidal activity of different extracts obtained from Cotula cinerea and Quercus lusitania var. infectoria galls. The hydroalcoholic extract of Cotula cinerea, acetonic extract and gallotanin of Quercus infectoria galls have presented high activity against Bulinus truncatus. The hydroalcoholic extract of Cotula cinerea was fractionated by chromatography on silica gel column. We have isolated two very active fractions at concentrations respectively of 52.5 and 27.5 ppm. PMID:9872015

  11. Ectopic expression of ubiquitin-conjugating enzyme gene from wild rice, OgUBC1, confers resistance against UV-B radiation and Botrytis infection in Arabidopsis thaliana

    SciTech Connect

    Jeon, En Hee; Pak, Jung Hun; Kim, Mi Jin; Kim, Hye Jeong; Shin, Sang Hyun; Lee, Jai Heon; Kim, Doh Hoon; Oh, Ju Sung; Oh, Boung-Jun; Jung, Ho Won; Chung, Young Soo

    2012-10-19

    Highlights: Black-Right-Pointing-Pointer We isolated a novel E2 ubiquitin-conjugating enzyme from leaves of wild rice plants. Black-Right-Pointing-Pointer The OgUBC1 was highly expressed in leaves treated with SA and UV-B radiation. Black-Right-Pointing-Pointer The recombinant OgUBC1 has an enzymatic activity of E2 in vitro. Black-Right-Pointing-Pointer The OgUBC1 could protect disruption of plant cells by UV-B radiation. Black-Right-Pointing-Pointer OgUBC1 confers disease resistance and UV-B tolerance in transgenic Arabidopsis plants. -- Abstract: A previously unidentified gene encoding ubiquitin-conjugating enzyme was isolated from leaves of wild rice plant treated with wounding and microbe-associated molecular patterns. The OgUBC1 gene was composed of 148 amino acids and contained a typical active site and 21 ubiquitin thioester intermediate interaction residues and 4 E3 interaction residues. Both exogenous application of salicylic acid and UV-B irradiation triggered expression of OgUBC1 in leaves of wild rice. Recombinant OgUBC1 proteins bound to ubiquitins in vitro, proposing that the protein might act as E2 enzyme in planta. Heterologous expression of the OgUBC1 in Arabidopsis thaliana protected plants from cellular damage caused by an excess of UV-B radiation. A stable expression of chalcone synthase gene was detected in leaves of OgUBC1-expressing Arabidopsis, resulting in producing higher amounts of anthocyanin than those in wild-type Col-0 plants. Additionally, both pathogenesis-related gene1 and 5 were transcribed in the transgenic Arabidopsis in the absence of pathogen infection. The OgUBC1-expressing plants were resistant to the infection of Botrytis cinerea. Taken together, we suggested that the OgUBC1 is involved in ubiquitination process important for cellular response against biotic and abiotic stresses in plants.

  12. Constitutive expression of Botrytis aclada laccase in Pichia pastoris.

    PubMed

    Kittl, Roman; Gonaus, Christoph; Pillei, Christian; Haltrich, Dietmar; Ludwig, Roland

    2012-01-01

    The heterologous expression of laccases is important for their large-scale production and genetic engineering--a prerequisite for industrial application. Pichia pastoris is the preferred expression host for fungal laccases. The recently cloned laccase from the ascomycete Botrytis aclada (BaLac) has been efficiently expressed in P. pastoris under the control of the inducible alcohol oxidase (AOX1) promoter. In this study, we compare these results to the constitutive expression in the same organism using the glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter. The results show that the amounts of BaLac produced with the GAP system (517 mgL(-1)) and the AOX1 system (495 mgL(-1)) are comparable. The constitutive expression is, however, faster, and the specific activity of BaLac in the culture supernatant is higher (41.3 Umg(-1) GAP, 14.2 Umg(-1) AOX1). In microtiter plates, the constitutive expression provides a clear advantage due to easy manipulation (simple medium, no methanol feeding) and fast enzyme production (high-throughput screening assays can already be performed after 48 h). PMID:22705842

  13. Constitutive expression of Botrytis aclada laccase in Pichia pastoris

    PubMed Central

    Kittl, Roman; Gonaus, Christoph; Pillei, Christian; Haltrich, Dietmar; Ludwig, Roland

    2012-01-01

    The heterologous expression of laccases is important for their large-scale production and genetic engineering—a prerequisite for industrial application. Pichia pastoris is the preferred expression host for fungal laccases. The recently cloned laccase from the ascomycete Botrytis aclada (BaLac) has been efficiently expressed in P. pastoris under the control of the inducible alcohol oxidase (AOX1) promoter. In this study, we compare these results to the constitutive expression in the same organism using the glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter. The results show that the amounts of BaLac produced with the GAP system (517 mgL-1) and the AOX1 system (495 mgL-1) are comparable. The constitutive expression is, however, faster, and the specific activity of BaLac in the culture supernatant is higher (41.3 Umg-1 GAP, 14.2 Umg-1 AOX1). In microtiter plates, the constitutive expression provides a clear advantage due to easy manipulation (simple medium, no methanol feeding) and fast enzyme production (high-throughput screening assays can already be performed after 48 h). PMID:22705842

  14. Novel Coprinopsis cinerea polyesterase that hydrolyzes cutin and suberin.

    PubMed

    Kontkanen, Hanna; Westerholm-Parvinen, Ann; Saloheimo, Markku; Bailey, Michael; Rättö, Marjaana; Mattila, Ismo; Mohsina, Marzia; Kalkkinen, Nisse; Nakari-Setälä, Tiina; Buchert, Johanna

    2009-04-01

    Three cutinase gene-like genes from the basidiomycete Coprinopsis cinerea (Coprinus cinereus) found with a similarity search were cloned and expressed in Trichoderma reesei under the control of an inducible cbh1 promoter. The selected transformants of all three polyesterase constructs showed activity with p-nitrophenylbutyrate, used as a model substrate. The most promising transformant of the cutinase CC1G_09668.1 gene construct was cultivated in a laboratory fermentor, with a production yield of 1.4 g liter(-l) purified protein. The expressed cutinase (CcCUT1) was purified to homogeneity by immobilized metal affinity chromatography exploiting a C-terminal His tag. The N terminus of the enzyme was found to be blocked. The molecular mass of the purified enzyme was determined to be around 18.8 kDa by mass spectrometry. CcCUT1 had higher activity on shorter (C(2) to C(10)) fatty acid esters of p-nitrophenol than on longer ones, and it also exhibited lipase activity. CcCUT1 had optimal activity between pH 7 and 8 but retained activity over a wide pH range. The enzyme retained 80% of its activity after 20 h of incubation at 50 degrees C, but residual activity decreased sharply at 60 degrees C. Microscopic analyses and determination of released hydrolysis products showed that the enzyme was able to depolymerize apple cutin and birch outer bark suberin. PMID:19201950

  15. Reproductive ecology of the green treefrog (Hyla cinerea) in Northwestern Florida

    USGS Publications Warehouse

    Gunzburger, M.S.

    2006-01-01

    Information on life-history traits is critical to understanding population dynamics of anurans. The objective of this study was to examine aspects of the reproductive ecology of Hyla cinerea in northwestern Florida. Four breeding localities in Leon County, Florida, were sampled over three seasons (2001-2003), amplexed pairs were found as early as 12 April and as late as 12 August. Egg clutches were counted from 51 amplexed pairs and adult size was measured in 43 pairs. Average clutch size was 1214 ?? 528 eggs (x- ?? SE, range = 359-2658). Female H. cinerea were slightly larger than males (x- ?? SE tibiofibula length = 24.9 ?? 2.2 and 24 ?? 1.7 mm, respectively). Female size was significantly positively correlated with clutch size and weakly correlated with size of the paired male. Comparison of the results of this study with previous research indicates that the reproductive ecology of H. cinerea varies across its geographic range.

  16. April showers bring May flowers…and May rains bring botrytis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As the old saying goes “April showers bring May flowers” and to the greenhouse production industry May brings “color” to the greenhouse in the form of flowers which are good for both spring sales and Botrytis. This should not come as anything new to the seasoned grower, but hopefully will serve as ...

  17. Studies on long-term preservation of dormant buds of Juglans cinerea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Juglans cinerea (butternut) is a deciduous tree native to the United States and Canada with oblong shaped nuts with an oily texture and a pleasant flavour. The species is threatened by a canker disease caused by the introduced fungus (Sirococcus clavigignenti-juglandacearum) which already eradicated...

  18. Differential predation on tadpoles influences the potential effects of hybridization between Hyla cinerea and Hyla gratiosa

    USGS Publications Warehouse

    Gunzburger, M.S.

    2005-01-01

    Long-term effects of hybridization and introgression are influenced by performance of hybrids in habitats of parental species. The treefrogs Hyla cinerea and Hyla gratiosa, which typically breed in permanent and temporary habitats, respectively, have occasionally hybridized throughout the Southeastern United States. To predict in which of the parental habitats effects of hybridization might be strongest, I performed experiments to evaluate predation on tadpoles of H. cinerea, H. gratiosa, and F1 hybrids with predators typical of the breeding habitats of the parental species. Hybrid tadpoles had lower survival with sunfish than odonate naiad (dragonfly) predators and tended to increase hiding behavior in response to sunfish predation. Tadpoles of H. gratiosa also had higher survival with odonates than sunfish, but H. cinerea had similar survival with both predator types. These results suggest that hybrids are most likely to survive and return to breed in temporary habitats used by H. gratiosa. Thus, hybridization and introgression might be more likely to have adverse effects on populations of H. gratiosa than H. cinerea. Copyright 2005 Society for the Study of Amphibians and Reptiles.

  19. Diversity of MHC DQB and DRB Genes in the Endangered Australian Sea Lion (Neophoca cinerea).

    PubMed

    Lau, Quintin; Chow, Natalie; Gray, Rachael; Gongora, Jaime; Higgins, Damien P

    2015-01-01

    Major histocompatibility complex (MHC) class II molecules have an important role in vertebrate adaptive immunity, being responsible for recognizing, binding, and presenting specific antigenic peptides to T lymphocytes. Here, we study the MHC class II DQB and DRB exon 2 genes of the Australian sea lion (Neophoca cinerea), an endangered pinniped species that experiences high pup mortality. Following characterization of N. cinerea DQB and DRB by molecular cloning, and evaluation of diversity in pups across 2 colonies using variant screening (n = 47), 3 DQB alleles and 10 DRB variants (including 1 pseudogene allele) were identified. The higher diversity at DRB relative to DQB is consistent with other studies in marine mammals. Despite overall lower MHC class II allelic diversity relative to some other pinniped species, we observed similar levels of nucleotide diversity and selection in N. cinerea. In addition, we provide support for recent divergence of MHC class II alleles. The characterization of MHC class II diversity in the Australian sea lion establishes a baseline for further investigation of associations with disease, including endemic hookworm infection, and contributes to the conservation management of this species. PMID:25908666

  20. Spatiotemporal relationships between disease development and airborne inoculum in unmanaged and managed Botrytis leaf blight epidemics.

    PubMed

    Carisse, O; Savary, S; Willocquet, L

    2008-01-01

    Comparatively little quantitative information is available on both the spatial and temporal relationships that develop between airborne inoculum and disease intensity during the course of aerially spread epidemics. Botrytis leaf blight and Botrytis squamosa airborne inoculum were analyzed over space and time during 2 years (2002 and 2004) in a nonprotected experimental field, using a 6 x 8 lattice of quadrats of 10 x 10 m each. A similar experiment was conducted in 2004 and 2006 in a commercial field managed for Botrytis leaf blight using a 5 x 5 lattice of quadrats of 25 x 25 m each. Each quadrat was monitored weekly for lesion density (LD) and aerial conidium concentration (ACC). The adjustment of the Taylor's power law showed that heterogeneity in both LD and ACC generally increased with increasing mean. Unmanaged epidemics were characterized in either year, with aggregation indices derived from SADIE (Spatial Analysis by Distance Indices). For LD, the aggregation indices suggested a random pattern of disease early in the season, followed by an aggregated pattern in the second part of the epidemic. The index of aggregation for ACC in 2002 was significantly greater than 1 at only one date, while it was significantly greater than 1 at most sampling dates in 2004. In both years and for both variables, positive trends in partial autocorrelation were observed mainly for a spatial lag of 1. In 2002, the overall pattern of partial autocorrelations over sampling dates was similar for LD and ACC with no significant partial autocorrelation during the first part of the epidemic, followed by a period with significant positive autocorrelation, and again no autocorrelation on the last three sampling dates. In 2004, there was no significant positive autocorrelation for LD at most sampling dates while for ACC, there was a fluctuation between significant and non-significant positive correlation over sampling dates. There was a significant spatial correlation between ACC at given

  1. GC/MS Analysis of the Essential Oil of Vernonia cinerea.

    PubMed

    Joshi, Rajesh K

    2015-07-01

    The hydro-distilled essential oil obtained from the roots of V. cinerea Less. (Asteraceae) was investigated by gas chromatography equipped with flame ionization detector (GC-FID) and gas chromatography coupled with mass spectrometry (GC/MS). Twenty-five constituents were identified, which represented 97.4% of the total oil. The major compounds were α-muurolene (30.7%), β-caryophyllene (9.6%), α-selinene (8.7%), cyperene (6.7%) and α-gurjunene (6.5%). The essential oil was dominated by sesquiterpene hydrocarbons (87.8%). PMID:26411040

  2. Population estimates of Hyla cinerea (Schneider) (Green Tree frog) in an urban environment

    USGS Publications Warehouse

    Pham, L.; Boudreaux, S.; Karhbet, S.; Price, B.; Ackleh, A.S.; Carter, J.; Pal, N.

    2007-01-01

    Hyla cinerea (Green Treefrog) is a common wetlands species in the southeastern US. To better understand its population dynamics, we followed a relatively isolated population of Green Treefrogs from June 2004 through October 2004 at a federal office complex in Lafayette, LA. Weekly, Green Treefrogs were caught, measured, marked with VIE tags, and released. The data were used to estimate population size. The time frame was split into two periods: before and after August 17, 2004. Before August 17, 2004, the average estimated population size was 143, and after August 24, 2005, this value jumped to 446, an increase possibly due to tadpoles metamorphosing into adults.

  3. A novel Botrytis species is associated with a newly emergent foliar disease in cultivated Hemerocallis.

    PubMed

    Grant-Downton, Robert T; Terhem, Razak B; Kapralov, Maxim V; Mehdi, Saher; Rodriguez-Enriquez, M Josefina; Gurr, Sarah J; van Kan, Jan A L; Dewey, Frances M

    2014-01-01

    Foliar tissue samples of cultivated daylilies (Hemerocallis hybrids) showing the symptoms of a newly emergent foliar disease known as 'spring sickness' were investigated for associated fungi. The cause(s) of this disease remain obscure. We isolated repeatedly a fungal species which proved to be member of the genus Botrytis, based on immunological tests. DNA sequence analysis of these isolates, using several different phyogenetically informative genes, indicated that they represent a new Botrytis species, most closely related to B. elliptica (lily blight, fire blight) which is a major pathogen of cultivated Lilium. The distinction of the isolates was confirmed by morphological analysis of asexual sporulating cultures. Pathogenicity tests on Hemerocallis tissues in vitro demonstrated that this new species was able to induce lesions and rapid tissue necrosis. Based on this data, we infer that this new species, described here as B. deweyae, is likely to be an important contributor to the development of 'spring sickness' symptoms. Pathogenesis may be promoted by developmental and environmental factors that favour assault by this necrotrophic pathogen. The emergence of this disease is suggested to have been triggered by breeding-related changes in cultivated hybrids, particularly the erosion of genetic diversity. Our investigation confirms that emergent plant diseases are important and deserve close monitoring, especially in intensively in-bred plants. PMID:24887415

  4. A Novel Botrytis Species Is Associated with a Newly Emergent Foliar Disease in Cultivated Hemerocallis

    PubMed Central

    Grant-Downton, Robert T.; Terhem, Razak B.; Kapralov, Maxim V.; Mehdi, Saher; Rodriguez-Enriquez, M. Josefina; Gurr, Sarah J.; van Kan, Jan A. L.; Dewey, Frances M.

    2014-01-01

    Foliar tissue samples of cultivated daylilies (Hemerocallis hybrids) showing the symptoms of a newly emergent foliar disease known as ‘spring sickness’ were investigated for associated fungi. The cause(s) of this disease remain obscure. We isolated repeatedly a fungal species which proved to be member of the genus Botrytis, based on immunological tests. DNA sequence analysis of these isolates, using several different phyogenetically informative genes, indicated that they represent a new Botrytis species, most closely related to B. elliptica (lily blight, fire blight) which is a major pathogen of cultivated Lilium. The distinction of the isolates was confirmed by morphological analysis of asexual sporulating cultures. Pathogenicity tests on Hemerocallis tissues in vitro demonstrated that this new species was able to induce lesions and rapid tissue necrosis. Based on this data, we infer that this new species, described here as B. deweyae, is likely to be an important contributor to the development of ‘spring sickness’ symptoms. Pathogenesis may be promoted by developmental and environmental factors that favour assault by this necrotrophic pathogen. The emergence of this disease is suggested to have been triggered by breeding-related changes in cultivated hybrids, particularly the erosion of genetic diversity. Our investigation confirms that emergent plant diseases are important and deserve close monitoring, especially in intensively in-bred plants. PMID:24887415

  5. FIRST REPORT OF BACTERIAL BLIGHT OF CAULIFLOWER (BRASSICA OLERACEA VAR. BOTRYTIS) CAUSED BY PSEUDOMONAS SYRINGAE PV. ALISALENSIS IN CALIFORNIA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A novel bacterial blight commercial romanesca (green) cauliflower (Brassica oleracea var. botrytis) was seen for the first time in coastal California in 2005. Koch’s postulates were completed and the etiology of the pathogen was determined. Characterization showed that pathogen was Pseudomonas syrin...

  6. Cryptic diversity within the choanoflagellate morphospecies complex Codosiga botrytis - phylogeny and morphology of ancient and modern isolates.

    PubMed

    Stoupin, Daniel; Kiss, Aron K; Arndt, Hartmut; Shatilovich, Anastasia V; Gilichinsky, David A; Nitsche, Frank

    2012-11-01

    Choanoflagellates are closely related to metazoans and fungi according to recent phylogenetic studies; therefore the systematics of these organisms is of particular interest. The choanoflagellate morphospecies Codosiga botrytis is the first described choanoflagellate, and is one of the most frequently reported choanoflagellate species. In this study we present phylogenetic and morphological data on eight different strains of Codosiga botrytis. Among these there are five ancient strains; these cultures have been established from up to 43,000 years old cysts from Siberian permafrost. We found that based on the variable V4 region of the small subunit (SSU) of the rDNA, all the investigated freshwater isolates of Codosiga botrytis, together with Sphaeroeca volvox, form a cluster at the base of all other choanoflagellate species. Moreover, the morphospecies described classically as Codosiga botrytis contains at least four different genotypes separated by considerably high genetic distance. All these 'cryptic species' have identical general morphology and cell structure. Strains have a similar life cycle with several different life forms and large morphological plasticity. For the first time we were able to establish cultures from cryo-conserved cysts of choanoflagellates. The ancient strains did not differ significantly in partial SSU rDNA from the modern ones. Besides, no biogeographically pattern could be established. This fact and the low genetic distances of some strains from remote locations support the distribution of dormant stages via air. PMID:22342133

  7. Strand-Specific RNA-Seq Analyses of Fruiting Body Development in Coprinopsis cinerea

    PubMed Central

    Muraguchi, Hajime; Umezawa, Kiwamu; Niikura, Mai; Yoshida, Makoto; Kozaki, Toshinori; Ishii, Kazuo; Sakai, Kiyota; Shimizu, Motoyuki; Nakahori, Kiyoshi; Sakamoto, Yuichi; Choi, Cindy; Ngan, Chew Yee; Lindquist, Eika; Lipzen, Anna; Tritt, Andrew; Haridas, Sajeet; Barry, Kerrie; Grigoriev, Igor V.; Pukkila, Patricia J.

    2015-01-01

    The basidiomycete fungus Coprinopsis cinerea is an important model system for multicellular development. Fruiting bodies of C. cinerea are typical mushrooms, which can be produced synchronously on defined media in the laboratory. To investigate the transcriptome in detail during fruiting body development, high-throughput sequencing (RNA-seq) was performed using cDNA libraries strand-specifically constructed from 13 points (stages/tissues) with two biological replicates. The reads were aligned to 14,245 predicted transcripts, and counted for forward and reverse transcripts. Differentially expressed genes (DEGs) between two adjacent points and between vegetative mycelium and each point were detected by Tag Count Comparison (TCC). To validate RNA-seq data, expression levels of selected genes were compared using RPKM values in RNA-seq data and qRT-PCR data, and DEGs detected in microarray data were examined in MA plots of RNA-seq data by TCC. We discuss events deduced from GO analysis of DEGs. In addition, we uncovered both transcription factor candidates and antisense transcripts that are likely to be involved in developmental regulation for fruiting. PMID:26510163

  8. Effects of multiple predator species on green treefrog (Hyla cinerea) tadpoles

    USGS Publications Warehouse

    Gunzburger, M.S.; Travis, J.

    2005-01-01

    Prey species that occur across a range of habitats may be exposed to variable communities of multiple predator species across habitats. Predicting the combined effects of multiple predators can be complex. Many experiments evaluating the effects of multiple predators on prey confound either variation in predator density with predator identity or variation in relative predator frequency with overall predation rates. We develop a new experimental design of factorial predator combinations that maintains a constant expected predation rate, under the null hypothesis of additive predator effects. We implement this design to evaluate the combined effects of three predator species (bass, aeshnid and libellulid odonate naiads) on mortality rate of a prey species, Hyla cinerea (Schneider, 1799) tadpoles, that occurs across a range of aquatic habitats. Two predator treatments (libellulid and aeshnid + libellulid) resulted in lower tadpole mortality than any of the other predator treatments. Variation in tadpole mortality across treatments was not related to coarse variation in microhabitat use, but was likely due to intraguild predation, which occurred in all predator treatments. Hyla cinerea tadpoles have constant, low survival values when exposed to many different combinations of predator species, and predation rate probably increases linearly with predator density.

  9. Strand-Specific RNA-Seq Analyses of Fruiting Body Development in Coprinopsis cinerea

    SciTech Connect

    Muraguchi, Hajime; Umezawa, Kiwamu; Niikura, Mai; Yoshida, Makoto; Kozaki, Toshinori; Ishii, Kazuo; Sakai, Kiyota; Shimizu, Motoyuki; Nakahori, Kiyoshi; Sakamoto, Yuichi; Choi, Cindy; Ngan, Chew Yee; Lindquist, Eika; Lipzen, Anna; Tritt, Andrew; Haridas, Sajeet; Barry, Kerrie; Grigoriev, Igor V.; Pukkila, Patricia J.

    2015-10-28

    We report that the basidiomycete fungus Coprinopsis cinerea is an important model system for multicellular development. Fruiting bodies of C. cinerea are typical mushrooms, which can be produced synchronously on defined media in the laboratory. To investigate the transcriptome in detail during fruiting body development, high-throughput sequencing (RNA-seq) was performed using cDNA libraries strand-specifically constructed from 13 points (stages/tissues) with two biological replicates. The reads were aligned to 14,245 predicted transcripts, and counted for forward and reverse transcripts. Differentially expressed genes (DEGs) between two adjacent points and between vegetative mycelium and each point were detected by Tag Count Comparison (TCC). To validate RNA-seq data, expression levels of selected genes were compared using RPKM values in RNA-seq data and qRT-PCR data, and DEGs detected in microarray data were examined in MA plots of RNA-seq data by TCC. We discuss events deduced from GO analysis of DEGs. In addition, we uncovered both transcription factor candidates and antisense transcripts that are likely to be involved in developmental regulation for fruiting.

  10. Strand-Specific RNA-Seq Analyses of Fruiting Body Development in Coprinopsis cinerea

    DOE PAGESBeta

    Muraguchi, Hajime; Umezawa, Kiwamu; Niikura, Mai; Yoshida, Makoto; Kozaki, Toshinori; Ishii, Kazuo; Sakai, Kiyota; Shimizu, Motoyuki; Nakahori, Kiyoshi; Sakamoto, Yuichi; et al

    2015-10-28

    We report that the basidiomycete fungus Coprinopsis cinerea is an important model system for multicellular development. Fruiting bodies of C. cinerea are typical mushrooms, which can be produced synchronously on defined media in the laboratory. To investigate the transcriptome in detail during fruiting body development, high-throughput sequencing (RNA-seq) was performed using cDNA libraries strand-specifically constructed from 13 points (stages/tissues) with two biological replicates. The reads were aligned to 14,245 predicted transcripts, and counted for forward and reverse transcripts. Differentially expressed genes (DEGs) between two adjacent points and between vegetative mycelium and each point were detected by Tag Count Comparison (TCC).more » To validate RNA-seq data, expression levels of selected genes were compared using RPKM values in RNA-seq data and qRT-PCR data, and DEGs detected in microarray data were examined in MA plots of RNA-seq data by TCC. We discuss events deduced from GO analysis of DEGs. In addition, we uncovered both transcription factor candidates and antisense transcripts that are likely to be involved in developmental regulation for fruiting.« less

  11. Neuroprotection of luteolin against methylmercury-induced toxicity in lobster cockroach Nauphoeta cinerea.

    PubMed

    Adedara, Isaac A; Rosemberg, Denis B; Souza, Diogo O; Farombi, Ebenezer O; Aschner, Michael; Rocha, Joao B T

    2016-03-01

    Luteolin (3', 4', 5, 7-tetrahydroxyflavone) is a polyphenolic compound found in foods of plant origin and has been reported to possess antioxidant and neuroprotective properties. However, there is dearth of information on the beneficial effects of luteolin on methylmercury (MeHg), a long-established neurotoxic compound in animals and humans. This study evaluated the effect of luteolin on MeHg-induced behavioral and biochemical deficits, using lobster cockroach Nauphoeta cinerea as an alternative and complementary animal model. The insects were exposed for 35 consecutive days to either MeHg alone (0.05 mg/g feed) or in combination with luteolin at 0.25, 0.5 and 1.0 mg/g feed. Locomotor behavior was assessed using video-tracking software during a 10-min trial in a novel arena and subsequently, biochemical analyses were carried out using the cockroaches' heads. Luteolin supplementation dose-dependently reversed the MeHg-induced locomotor deficits and enhanced the exploratory profiles of MeHg-exposed cockroaches as confirmed by track and occupancy plot analyses. Luteolin reversed the MeHg-induced acetylcholinesterase activity inhibition, decreased dichlorofluorescein oxidation and lipid peroxidation levels, but increased total thiol level and catalase and glutathione S-transferase activities in the treated cockroaches. In conclusion, luteolin prevented oxidative stress indices and neurobehavioral deficits in a Nauphoeta cinerea model of MeHg toxicity. PMID:26905302

  12. Neurobehavioral and biochemical changes in Nauphoeta cinerea following dietary exposure to chlorpyrifos.

    PubMed

    Adedara, Isaac A; Rosemberg, Denis B; de Souza, Diego; Farombi, Ebenezer O; Aschner, Michael; Souza, Diogo O; Rocha, Joao B T

    2016-06-01

    The present study aimed to increase our understanding about the mode of toxic action of organophosphate pesticides in insects by evaluating the biochemical and neurobehavioral characteristics in Nauphoeta cinerea exposed to chlorpyrifos (CPF)-contaminated diet. The insects were exposed for 35 consecutive days to CPF at 0.078, 0.15625, 0.3125 and 0.625μg/g feed. Locomotor behavior was assessed for a 10-min trial in a novel arena and subsequently, biochemical analyses were carried out using the cockroaches' heads. In comparison to control, CPF-exposed cockroaches showed significant decreases in the total distance traveled, body rotation, turn angle and meandering, along with significant increase in the number of falls, time and episodes of immobility. The marked decrease in the exploratory profiles of CPF-exposed cockroaches was confirmed by track plots, whereas occupancy plot analyses showed a progressive dispersion at 0.15625μg/g feed group. Moreover, the heads of CPF-exposed cockroaches showed marked decrease in acetylcholinesterase activity and antioxidant status with concomitant significant elevation in dichlorofluorescein oxidation and lipid peroxidation levels in CPF-treated cockroaches. Gas Chromatography-Mass Spectrometry analyses revealed bioaccumulation of CPF in cockroaches exposed to concentrations above 0.078μg/g feed. The findings from this investigation showed N. cinerea as a value model organism for the risk assessment of environmental organophosphate contamination in insects. PMID:27155480

  13. Identification of a novel nematotoxic protein by challenging the model mushroom Coprinopsis cinerea with a fungivorous nematode

    DOE PAGESBeta

    Plaza, David Fernando; Schmieder, Stefanie Sofia; Lipzen, Anna; Lindquist, Erika; Künzler, Markus

    2015-11-19

    The dung of herbivores, the natural habitat of the model mushroom Coprinopsis cinerea, is a nutrient-rich but also very competitive environment for a saprophytic fungus. Previously we showed that C. cinerea expresses constitutive, tissue-specific armories against antagonists such as animal predators and bacterial competitors. In order to dissect the inducible armories against such antagonists, we sequenced the poly(A)-positive transcriptome of C. cinerea vegetative mycelium upon challenge with fungivorous and bacterivorous nematodes, Gram-negative and Gram-positive bacteria and mechanical damage. As a response to the fungivorous nematode Aphelenchus avenae, C. cinerea was found to specifically induce the transcription of several genes encodingmore » previously characterized nematotoxic lectins. In addition, a previously not characterized gene encoding a cytoplasmic protein with several predicted Ricin B-fold domains, was found to be strongly up-regulated under this condition. Functional analysis of the recombinant protein revealed a high toxicity towards the bacterivorous nematode Caenorhabditis elegans. Challenge of the mycelium with A. avenae also lead to the induction of several genes encoding putative antibacterial proteins. Some of these genes were also induced upon challenge of the mycelium with the bacteria Escherichia coli and Bacillus subtilis. Lastly, these results suggest that fungi have the ability to induce specific innate defense responses similar to plants and animals.« less

  14. Identification of a novel nematotoxic protein by challenging the model mushroom Coprinopsis cinerea with a fungivorous nematode

    SciTech Connect

    Plaza, David Fernando; Schmieder, Stefanie Sofia; Lipzen, Anna; Lindquist, Erika; Künzler, Markus

    2015-11-19

    The dung of herbivores, the natural habitat of the model mushroom Coprinopsis cinerea, is a nutrient-rich but also very competitive environment for a saprophytic fungus. Previously we showed that C. cinerea expresses constitutive, tissue-specific armories against antagonists such as animal predators and bacterial competitors. In order to dissect the inducible armories against such antagonists, we sequenced the poly(A)-positive transcriptome of C. cinerea vegetative mycelium upon challenge with fungivorous and bacterivorous nematodes, Gram-negative and Gram-positive bacteria and mechanical damage. As a response to the fungivorous nematode Aphelenchus avenae, C. cinerea was found to specifically induce the transcription of several genes encoding previously characterized nematotoxic lectins. In addition, a previously not characterized gene encoding a cytoplasmic protein with several predicted Ricin B-fold domains, was found to be strongly up-regulated under this condition. Functional analysis of the recombinant protein revealed a high toxicity towards the bacterivorous nematode Caenorhabditis elegans. Challenge of the mycelium with A. avenae also lead to the induction of several genes encoding putative antibacterial proteins. Some of these genes were also induced upon challenge of the mycelium with the bacteria Escherichia coli and Bacillus subtilis. Lastly, these results suggest that fungi have the ability to induce specific innate defense responses similar to plants and animals.

  15. Identification of a Novel Nematotoxic Protein by Challenging the Model Mushroom Coprinopsis cinerea with a Fungivorous Nematode

    PubMed Central

    Plaza, David Fernando; Schmieder, Stefanie Sofia; Lipzen, Anna; Lindquist, Erika; Künzler, Markus

    2015-01-01

    The dung of herbivores, the natural habitat of the model mushroom Coprinopsis cinerea, is a nutrient-rich but also very competitive environment for a saprophytic fungus. We showed previously that C. cinerea expresses constitutive, tissue-specific armories against antagonists such as animal predators and bacterial competitors. In order to dissect the inducible armories against such antagonists, we sequenced the poly(A)-positive transcriptome of C. cinerea vegetative mycelium upon challenge with fungivorous and bacterivorous nematodes, Gram-negative and Gram-positive bacteria and mechanical damage. As a response to the fungivorous nematode Aphelenchus avenae, C. cinerea was found to specifically induce the transcription of several genes encoding previously characterized nematotoxic lectins. In addition, a previously not characterized gene encoding a cytoplasmic protein with several predicted Ricin B-fold domains, was found to be strongly upregulated under this condition. Functional analysis of the recombinant protein revealed a high toxicity toward the bacterivorous nematode Caenorhabditis elegans. Challenge of the mycelium with A. avenae also lead to the induction of several genes encoding putative antibacterial proteins. Some of these genes were also induced upon challenge of the mycelium with the bacteria Escherichia coli and Bacillus subtilis. These results suggest that fungi have the ability to induce specific innate defense responses similar to plants and animals. PMID:26585824

  16. Identification of a Novel Nematotoxic Protein by Challenging the Model Mushroom Coprinopsis cinerea with a Fungivorous Nematode.

    PubMed

    Plaza, David Fernando; Schmieder, Stefanie Sofia; Lipzen, Anna; Lindquist, Erika; Künzler, Markus

    2016-01-01

    The dung of herbivores, the natural habitat of the model mushroom Coprinopsis cinerea, is a nutrient-rich but also very competitive environment for a saprophytic fungus. We showed previously that C. cinerea expresses constitutive, tissue-specific armories against antagonists such as animal predators and bacterial competitors. In order to dissect the inducible armories against such antagonists, we sequenced the poly(A)-positive transcriptome of C. cinerea vegetative mycelium upon challenge with fungivorous and bacterivorous nematodes, Gram-negative and Gram-positive bacteria and mechanical damage. As a response to the fungivorous nematode Aphelenchus avenae, C. cinerea was found to specifically induce the transcription of several genes encoding previously characterized nematotoxic lectins. In addition, a previously not characterized gene encoding a cytoplasmic protein with several predicted Ricin B-fold domains, was found to be strongly upregulated under this condition. Functional analysis of the recombinant protein revealed a high toxicity toward the bacterivorous nematode Caenorhabditis elegans. Challenge of the mycelium with A. avenae also lead to the induction of several genes encoding putative antibacterial proteins. Some of these genes were also induced upon challenge of the mycelium with the bacteria Escherichia coli and Bacillus subtilis. These results suggest that fungi have the ability to induce specific innate defense responses similar to plants and animals. PMID:26585824

  17. Incorporation of copper ions into crystals of T2 copper-depleted laccase from Botrytis aclada

    PubMed Central

    Osipov, E. M.; Polyakov, K. M.; Tikhonova, T. V.; Kittl, R.; Dorovatovskii, P.V.; Shleev, S. V.; Popov, V. O.; Ludwig, R.

    2015-01-01

    Laccases belong to the class of multicopper oxidases catalyzing the oxidation of phenols accompanied by the reduction of molecular oxygen to water without the formation of hydrogen peroxide. The activity of laccases depends on the number of Cu atoms per enzyme molecule. The structure of type 2 copper-depleted laccase from Botrytis aclada has been solved previously. With the aim of obtaining the structure of the native form of the enzyme, crystals of the depleted laccase were soaked in Cu+- and Cu2+-containing solutions. Copper ions were found to be incorporated into the active site only when Cu+ was used. A comparative analysis of the native and depleted forms of the enzymes was performed. PMID:26625287

  18. Incorporation of copper ions into crystals of T2 copper-depleted laccase from Botrytis aclada.

    PubMed

    Osipov, E M; Polyakov, K M; Tikhonova, T V; Kittl, R; Dorovatovskii, P V; Shleev, S V; Popov, V O; Ludwig, R

    2015-12-01

    Laccases belong to the class of multicopper oxidases catalyzing the oxidation of phenols accompanied by the reduction of molecular oxygen to water without the formation of hydrogen peroxide. The activity of laccases depends on the number of Cu atoms per enzyme molecule. The structure of type 2 copper-depleted laccase from Botrytis aclada has been solved previously. With the aim of obtaining the structure of the native form of the enzyme, crystals of the depleted laccase were soaked in Cu(+)- and Cu(2+)-containing solutions. Copper ions were found to be incorporated into the active site only when Cu(+) was used. A comparative analysis of the native and depleted forms of the enzymes was performed. PMID:26625287

  19. Diseases and Their Management

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Important diseases and their management practices of lentil were reviewed. The diseases reveiwed include Ascochyta blight (Ascochyta lentis), Anthracnose (Colletotrichum truncatum), White mold (Sclerotinia sclerotiorum), rust (Uromyces viciae-fabae), Botrytis gray mold (Botrytis cinerea and B. faba...

  20. Foreword: Special issue on fungal grapevine diseases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An impressively large proportion of fungicides applied in European, North American and Australian agriculture has been used to manage grapevine powdery mildew (Erysiphe necator), grapevine downy mildew (Plasmopara viticola), and botrytis bunch rot (Botrytis cinerea). These fungal and oomycetous plan...

  1. Discontinuous gas exchange exhibition is a heritable trait in speckled cockroaches Nauphoeta cinerea.

    PubMed

    Schimpf, N G; Matthews, P G D; White, C R

    2013-07-01

    The regulation of insect respiratory gas exchange has long been an area of interest. In particular, the reason why insects from at least five orders exhibit patterns of gas exchange that include regular periods of spiracular closure has been the source of much controversy. Three adaptive hypotheses propose that these discontinuous gas-exchange cycles (DGCs) evolved to either limit water loss across respiratory surfaces, facilitate gas exchange in underground environments or to limit oxidative damage. It is possible that DGCs evolved independently multiple times and for different reasons, but for DGCs to be a plausible target for natural selection, they must be heritable and confer a fitness benefit. In a previous study of cockroaches Nauphoeta cinerea, we demonstrated that DGCs are repeatable and extend survival under food and water restriction. Here, we show for the first time that DGCs are heritable, suggesting that they are a plausible target for natural selection. PMID:23662792

  2. Bovine tuberculosis in an Asian small-clawed otter (Aonyx cinerea) in the Republic of Korea.

    PubMed

    Lee, Hyunkyoung; Kim, Jae-Myung; Jang, Yunho; Lee, Kyunghyun; Baek, Kanghyun; Lee, Boram; Kim, Ha-Young; Lee, Myoung-Heon; Ryoo, Soyoon; Bae, You-Chan; Choi, Eun-Jin; So, ByungJae

    2015-09-01

    Bovine tuberculosis caused by Mycobacterium bovis has a wide range of hosts including cattle and humans, but its incidence in otters is very rare. Our report describes a case of bovine tuberculosis in an Asian small-clawed otter (Aonyx cinerea). A deceased female otter ~2-3 years of age that was raised in an aquarium was submitted to the Animal and Plant Quarantine Agency (Anyang, Republic of Korea) for autopsy in June 2013. Following gross pathological examination, many white nodules were observed in the lungs and mesentery. The nodules showed central necrosis infiltrated with lymphocytes and macrophages and surrounded by fibrous tissue. Acid-fast bacteria were detected in the necrotic foci, but no fungi were observed. Molecular analysis led to the detection of M. bovis, which is identified in otters in some European countries such as Spain and France. PMID:26289719

  3. Sexually Dimorphic Effects of Melatonin on Brain Arginine Vasotocin Immunoreactivity in Green Treefrogs (Hyla cinerea)

    PubMed Central

    Lutterschmidt, Deborah I.; Wilczynski, Walter

    2012-01-01

    Arginine vasotocin (AVT) and its mammalian homologue, arginine vasopressin (AVP), regulate a variety of social and reproductive behaviors, often with complex species-, sex-, and context-dependent effects. Despite extensive evidence documenting seasonal variation in brain AVT/AVP, relatively few studies have investigated the environmental and/or hormonal factors mediating these seasonal changes. In the present study, we investigated whether the pineal hormone melatonin alters brain AVT immunoreactivity in green treefrogs (Hyla cinerea). Reproductively active male and female frogs were collected during the summer breeding season and a melatonin-filled or blank silastic capsule was surgically implanted subcutaneously. The duration of hormone treatment was 4 weeks, at which time frogs were euthanized and the brains and blood collected and processed for AVT immunohistochemistry and steroid hormone assay. We quantified AVT-immunoreactive (AVT-ir) cell bodies in the nucleus accumbens (NAcc), caudal striatum and amygdala (AMG), anterior preoptic area (POA), suprachiasmatic nucleus (SCN), and infundibular region of the ventral hypothalamus (VH). Sex differences in AVT-ir cell number were observed in all brain regions except the anterior POA and VH, with males having more AVT-ir cells than females in the NAcc, AMG, and SCN. Brain AVT was sensitive to melatonin signaling during the breeding season, and the effects of melatonin varied significantly with both region and sex. Treatment with melatonin decreased AVT immunoreactivity in both the NAcc and SCN in male H. cinerea. In contrast, brain AVT was relatively insensitive to melatonin signaling in females, indicating that the regulation of the AVT/AVP neuropeptide system by melatonin may be sexually dimorphic. Finally, melatonin did not significantly influence testosterone or estradiol concentrations of male or female frogs, respectively, suggesting that the effects of melatonin on AVT immunoreactivity are independent of

  4. Purification, characterization and physiological significance of a chitinase from the pilei of Coprinopsis cinerea fruiting bodies.

    PubMed

    Zhou, Yajun; Kang, Liqin; Niu, Xin; Wang, Jun; Liu, Zhonghua; Yuan, Sheng

    2016-06-01

    We purified a chitinase from pilei extractions of Coprinopsis cinerea fruiting bodies by ammonium sulfate precipitation and CM Sepharose cation exchange chromatography. MALDI-TOF/TOF MS analysis characterized this purified chitinase as a putative class V chitinase, ChiB1. ChiB1 hydrolyzed colloidal chitin and chitosan, whereas it did not hydrolyze chitin powder. ChiB1 cleaved only pNP-(GlcNAc)2, rather than pNP-GlcNAc or pNP-(Glc-NAc)3, to release nitrophenol. ChiB1 preferably and progressively released (GlcNAc)2 from (GlcNAc)6 and digested (GlcNAc)6 to two molecules of (GlcNAc)3 in a small proportion, but did not split (GlcNAc)2, so it is an exochitinase. ChiB1 has an optimum temperature range of 35°C to 40°C and an optimum pH of 5.0. ChiB1 exhibited Km and Vmax values of 2.63 mg ml(-1) and 2.31 μmol min(-1) mg protein(-1) for colloidal chitin, respectively. The ChiB1 gene, along with another putative endochitinase (class III chitinase gene), was expressed dominantly among eight predicted chitinase genes in the genome, and its expression level increased with the maturation of fruiting bodies. ChiB1 incubation released a large amount of soluble β-glucan fractions from alkali-insoluble cell wall fractions of C. cinerea fruiting bodies, thereby it may promote the degradation of cell walls in synergy with the β-1,3-glucanases during pileus autolysis. PMID:27190145

  5. Is the lobster cockroach Nauphoeta cinerea a valuable model for evaluating mercury induced oxidative stress?

    PubMed

    Rodrigues, N R; Nunes, M E M; Silva, D G C; Zemolin, A P P; Meinerz, D F; Cruz, L C; Pereira, A B; Rocha, J B T; Posser, T; Franco, J L

    2013-08-01

    Organic and inorganic forms of mercury are highly neurotoxic environmental contaminants. The exact mechanisms involved in mercury neurotoxicity are still unclear. Oxidative stress appears to play central role in this process. In this study, we aimed to validate an insect-based model for the investigation of oxidative stress during mercury poisoning of lobster cockroach Nauphoeta cinerea. The advantages of using insects in basic toxicological studies include the easier handling, rapid proliferation/growing and absence of ethical issues, comparing to rodent-based models. Insects received solutions of HgCl2 (10, 20 and 40mgL(-1) in drinking water) for 7d. 24h after mercury exposure, animals were euthanized and head tissue samples were prepared for oxidative stress related biochemical determinations. Mercury exposure caused a concentration dependent decrease in survival rate. Cholinesterase activity was unchanged. Catalase activity was substantially impaired after mercury treatment 40mgL(-1). Likewise, GST had a significant decrease, comparing to control. Peroxidase and thioredoxin reductase activity was inhibited at concentrations of 20mgL(-1) and 40mgL(-1) comparing to control. These results were accompanied by decreased GSH levels and increased hydroperoxide and TBARS formation. In conclusion, our results show that mercuric compounds are able to induce oxidative stress signs in insect by modulating survival rate as well as inducing impairments on important antioxidant systems. In addition, our data demonstrates for the first time that Nauphoeta cinerea represents an interesting animal model to investigate mercury toxicity and indicates that the GSH and thioredoxin antioxidant systems plays central role in Hg induced toxicity in insects. PMID:23466093

  6. Insights into evolution of multicellular fungi from the assembled chromosomes of the mushroom Coprinopsis cinerea (Coprinus cinereus)

    PubMed Central

    Stajich, Jason E.; Wilke, Sarah K.; Ahrén, Dag; Au, Chun Hang; Birren, Bruce W.; Borodovsky, Mark; Burns, Claire; Canbäck, Björn; Casselton, Lorna A.; Cheng, C.K.; Dietrich, Fred S.; Fargo, David C.; Farman, Mark L.; Gathman, Allen C.; Goldberg, Jonathan; Guigó, Roderic; Hoegger, Patrick J.; Hooker, James B.; Huggins, Ashleigh; James, Timothy Y.; Kamada, Takashi; Kilaru, Sreedhar; Kodira, Chinnapa; Kües, Ursula; Kupfer, Doris; Kwan, H.S.; Lomsadze, Alexandre; Li, Weixi; Lilly, Walt W.; Ma, Li-Jun; Mackey, Aaron J.; Manning, Gerard; Martin, Francis; Muraguchi, Hajime; Natvig, Donald O.; Palmerini, Heather; Ramesh, Marilee A.; Rehmeyer, Cathy J.; Roe, Bruce A.; Shenoy, Narmada; Stanke, Mario; Ter-Hovhannisyan, Vardges; Tunlid, Anders; Velagapudi, Rajesh; Vision, Todd J.; Zeng, Qiandong; Zolan, Miriam E.; Pukkila, Patricia J.

    2010-01-01

    The mushroom Coprinopsis cinerea is a classic experimental model for multicellular development in fungi because it grows on defined media, completes its life cycle in 2 weeks, produces some 108 synchronized meiocytes, and can be manipulated at all stages in development by mutation and transformation. The 37-megabase genome of C. cinerea was sequenced and assembled into 13 chromosomes. Meiotic recombination rates vary greatly along the chromosomes, and retrotransposons are absent in large regions of the genome with low levels of meiotic recombination. Single-copy genes with identifiable orthologs in other basidiomycetes are predominant in low-recombination regions of the chromosome. In contrast, paralogous multicopy genes are found in the highly recombining regions, including a large family of protein kinases (FunK1) unique to multicellular fungi. Analyses of P450 and hydrophobin gene families confirmed that local gene duplications drive the expansions of paralogous copies and the expansions occur in independent lineages of Agaricomycotina fungi. Gene-expression patterns from microarrays were used to dissect the transcriptional program of dikaryon formation (mating). Several members of the FunK1 kinase family are differentially regulated during sexual morphogenesis, and coordinate regulation of adjacent duplications is rare. The genomes of C. cinerea and Laccaria bicolor, a symbiotic basidiomycete, share extensive regions of synteny. The largest syntenic blocks occur in regions with low meiotic recombination rates, no transposable elements, and tight gene spacing, where orthologous single-copy genes are overrepresented. The chromosome assembly of C. cinerea is an essential resource in understanding the evolution of multicellularity in the fungi. PMID:20547848

  7. Incorporation of copper ions into crystals of T2 copper-depleted laccase from Botrytis aclada

    SciTech Connect

    Osipov, E. M.; Polyakov, K. M.; Tikhonova, T. V.; Kittl, R.; Dorovatovskii, P.V.; Shleev, S. V.; Popov, V. O.; Ludwig, R.

    2015-11-18

    The restoration of the native form of laccase from B. aclada from the type 2 copper-depleted form of the enzyme was investigated. Copper ions were found to be incorporated into the active site after soaking the depleted enzyme in a Cu{sup +}-containing solution. Laccases belong to the class of multicopper oxidases catalyzing the oxidation of phenols accompanied by the reduction of molecular oxygen to water without the formation of hydrogen peroxide. The activity of laccases depends on the number of Cu atoms per enzyme molecule. The structure of type 2 copper-depleted laccase from Botrytis aclada has been solved previously. With the aim of obtaining the structure of the native form of the enzyme, crystals of the depleted laccase were soaked in Cu{sup +}- and Cu{sup 2+}-containing solutions. Copper ions were found to be incorporated into the active site only when Cu{sup +} was used. A comparative analysis of the native and depleted forms of the enzymes was performed.

  8. Antenna contact and agonism in the male lobster cockroach, Nauphoeta cinerea.

    PubMed

    Chou, Szu-Ying; Huang, Zachary Y; Chen, Shu-Chun; Yang, Rou-Ling; Kou, Rong

    2007-08-01

    On any given day, about 35% of 80- to 85-day-old socially naïve male (SNM) lobster cockroaches (Nauphoeta cinerea) spontaneously adopted an aggressive posture (AP) without encountering another male [spontaneous AP (SAP)]. Although SAP SNMs showed significantly higher release of the pheromone 3-hydroxy-2-butanone (3H-2B) than non-SAP SNMs, there was no significant difference in hemolymph juvenile hormone (JH) III titer. When different body parts were tested for induction of the attack behavior, the antenna was found to be the most effective. After 1 min of contact with an antenna from another SAP SNM, attack behavior was induced in 100% of SAP and 76.2% of non-SAP SNMs, and the JH III titer was significantly increased in all responders. Among the non-SAP SNMs, the JH III titer before antenna contact was significantly lower in the non-responders than in the responders, and, although the JH III increase induced by 1 min antenna contact was similar between responders and non-responders, the final JH III titer of the non-responders was significantly lower. A similar attack response, JH III titer change, and 3H-2B release were seen when the individual's own antenna was used. After 5 min of contact with an antenna from another SAP SNM, attack behavior was induced in 100% of SAP and 82% of non-SAP SNMs; in the former, 3H-2B release was similar before and after antenna contact, but the JH III titer was significantly increased after antenna contact, while, in the latter, both 3H-2B release and JH III titer were significantly increased after antenna contact. Among the non-SAP SNMs, JH III titer in the non-responders was not elevated after 5 min antenna contact, and was significantly lower than that in the responders. A pentane-washed antenna did not induce attack behavior or increase the hemolymph JH III titer, and a pentane-washed antenna coated with 3H-2B also failed to induce attack behavior. These results indicate that N. cinerea male-male agonistic interactions, to

  9. Oxygen-induced plasticity in tracheal morphology and discontinuous gas exchange cycles in cockroaches Nauphoeta cinerea.

    PubMed

    Bartrim, Hamish; Matthews, Philip G D; Lemon, Sussan; White, Craig R

    2014-12-01

    The function and mechanism underlying discontinuous gas exchange in terrestrial arthropods continues to be debated. Three adaptive hypotheses have been proposed to explain the evolutionary origin or maintenance of discontinuous gas exchange cycles (DGCs), which may have evolved to reduce respiratory water loss, facilitate gas exchange in high CO2 and low O2 micro-environments, or to ameliorate potential damage as a result of oversupply of O2. None of these hypotheses have unequivocal support, and several non-adaptive hypotheses have also been proposed. In the present study, we reared cockroaches Nauphoeta cinerea in selected levels of O2 throughout development, and examined how this affected growth rate, tracheal morphology and patterns of gas exchange. O2 level in the rearing environment caused significant changes in tracheal morphology and the exhibition of DGCs, but the direction of these effects was inconsistent with all three adaptive hypotheses: water loss was not associated with DGC length, cockroaches grew fastest in hyperoxia, and DGCs exhibited by cockroaches reared in normoxia were shorter than those exhibited by cockroaches reared in hypoxia or hyperoxia. PMID:25378216

  10. Simultaneous sequencing of oxidized methylcytosines produced by TET/JBP dioxygenases in Coprinopsis cinerea

    PubMed Central

    Chavez, Lukas; Huang, Yun; Luong, Khai; Agarwal, Suneet; Iyer, Lakshminarayan M.; Pastor, William A.; Hench, Virginia K.; Frazier-Bowers, Sylvia A.; Korol, Evgenia; Liu, Shuo; Tahiliani, Mamta; Wang, Yinsheng; Clark, Tyson A.; Korlach, Jonas; Pukkila, Patricia J.; Aravind, L.; Rao, Anjana

    2014-01-01

    TET/JBP enzymes oxidize 5-methylpyrimidines in DNA. In mammals, the oxidized methylcytosines (oxi-mCs) function as epigenetic marks and likely intermediates in DNA demethylation. Here we present a method based on diglucosylation of 5-hydroxymethylcytosine (5hmC) to simultaneously map 5hmC, 5-formylcytosine, and 5-carboxylcytosine at near–base-pair resolution. We have used the method to map the distribution of oxi-mC across the genome of Coprinopsis cinerea, a basidiomycete that encodes 47 TET/JBP paralogs in a previously unidentified class of DNA transposons. Like 5-methylcytosine residues from which they are derived, oxi-mC modifications are enriched at centromeres, TET/JBP transposons, and multicopy paralogous genes that are not expressed, but rarely mark genes whose expression changes between two developmental stages. Our study provides evidence for the emergence of an epigenetic regulatory system through recruitment of selfish elements in a eukaryotic lineage, and describes a method to map all three different species of oxi-mCs simultaneously. PMID:25406324

  11. Dip listening or modulation masking? Call recognition by green treefrogs (Hyla cinerea) in temporally fluctuating noise.

    PubMed

    Vélez, Alejandro; Höbel, Gerlinde; Gordon, Noah M; Bee, Mark A

    2012-12-01

    Despite the importance of perceptually separating signals from background noise, we still know little about how nonhuman animals solve this problem. Dip listening, an ability to catch meaningful 'acoustic glimpses' of a target signal when fluctuating background noise levels momentarily drop, constitutes one possible solution. Amplitude-modulated noises, however, can sometimes impair signal recognition through a process known as modulation masking. We asked whether fluctuating noise simulating a breeding chorus affects the ability of female green treefrogs (Hyla cinerea) to recognize male advertisement calls. Our analysis of recordings of the sounds of green treefrog choruses reveal that their levels fluctuate primarily at rates below 10 Hz. In laboratory phonotaxis tests, we found no evidence for dip listening or modulation masking. Mean signal recognition thresholds in the presence of fluctuating chorus-like noises were never statistically different from those in the presence of a non-fluctuating control. An analysis of statistical effects sizes indicates that masker fluctuation rates, and the presence versus absence of fluctuations, had negligible effects on subject behavior. Together, our results suggest that females listening in natural settings should receive no benefits, nor experience any additional constraints, as a result of level fluctuations in the soundscape of green treefrog choruses. PMID:23069882

  12. Morphological and molecular methods to identify butternut (Juglans cinerea) and butternut hybrids: relevance to butternut conservation.

    PubMed

    Ross-Davis, Amy; Huang, Zhonglian; McKenna, James; Ostry, Michael; Woeste, Keith

    2008-07-01

    Butternut (Juglans cinerea L.) is a native, cold-tolerant, hard-mast species formerly valued for its nuts and wood, which is now endangered. The most immediate threat to butternut restoration is the spread of butternut canker disease, caused by the exotic fungus Sirococcus clavigignenti-juglandacearum Nair, Kostichka & Kuntz. Other threats include the hybridization of butternut with the exotic Japanese walnut (Juglans ailantifolia Carr.) and poor regeneration. The hybrids, known as buartnuts, are vegetatively vigorous, highly fecund, more resistant than butternut to butternut canker disease and difficult to identify. We review the vegetative and reproductive morphological traits that distinguish butternut from hybrids and identify those that can be used by field biologists to separate the taxa. No single trait was sufficient to separate butternut from hybrids, but pith color, lenticel size, shape and abundance, and the presence or absence of a notch in the upper margin of leaf scars, can be used in combination with other traits to identify butternuts and exclude most hybrids. In at least one butternut population, reduced symptoms of butternut canker disease were significantly associated with a dark barked phenotype. We also describe two randomly amplified polymorphic DNA (RAPD) markers that differentiate butternuts from hybrids based on DNA polymorphism. Together, these results should assist in the identification and testing of non-hybrid butternut for breeding and reintroduction of the species to its former habitats. PMID:18450577

  13. Uncinaria sanguinis sp. n. (Nematoda: Ancylostomatidae) from the endangered Australian sea lion, Neophoca cinerea (Carnivora: Otariidae).

    PubMed

    Marcus, Alan D; Higgins, Damien P; Slapeta, Jan; Gray, Rachael

    2014-06-01

    This study investigates the identity of hookworms parasitising the Australian sea lion, Neophoca cinerea (Péron), from three colonies in South Australia, Australia. The Australian sea lion is at risk of extinction because its population is small and genetically fragmented. Using morphological and molecular techniques, we describe a single novel species, Uncinaria sanguinis sp. n. (Nematoda: Ancylostomatidae). The new species is most similar to hookworms also parasitic in otariid hosts, Uncinaria lucasi Stiles, 1901 and Uncinaria hamiltoni Baylis, 1933. Comparative morphometrics offered limited utility for distinguishing between species within this genus whilst morphological features and differences in nuclear ribosomal DNA sequences delineated U. sanguinis sp. n. from named congeners. Male specimens of U. sanguinis sp. n. differ from U. lucasi and U. hamiltoni by relatively shorter anterolateral and externodorsal rays, respectively, and from other congeners by the relative lengths and angulations of bursal rays, and in the shape of the spicules. Female specimens of U. sanguinis sp. n. are differentiated from Uncinaria spp. parasitic in terrestrial mammals by differences in vulval anatomy and the larger size of their eggs, although are morphologically indistinguishable from U. lucasi and U. hamiltoni. Molecular techniques clearly delimited U. sanguinis sp. n. as a distinct novel species. Obtaining baseline data on the parasites of wildlife hosts is important for the investigation of disease and the effective implementation and monitoring of conservation management. PMID:25065131

  14. Purification and characterization of a laccase from Coprinopsis cinerea in Pichia pastoris.

    PubMed

    Wang, Bo; Wang, Lijuan; Lin, Yaqiu; Han, Qing; Han, Jing; Gao, Jianjie; Tian, Yongsheng; Zhao, Wei; Peng, Rihe; Yao, Quanhong

    2014-04-01

    A modified laccase gene, CcLCC6, from Coprinopsis cinerea was chemically synthesized according to the yeast codon bias and expressed in Pichia pastoris. The main properties of laccase, effects of ions and inhibitors, and optimal condition for decolouring malachite green (MG) were investigated in this study. The optimal pH level and temperature of laccase are 3.0 and 40 °C, respectively. The metal ions Mn²⁺, Zn²⁺, Fe³⁺ and Al³⁺ could inhibit laccase activity, as well as 1 mM of sodium dodecyl sulphate and sodium thiosulphate. 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), as a mediator, was necessary in decolorizing MG. The optimal pH and temperature for MG decolorization were 3.0 and 50 °C, respectively. Approximately 0.02 μM recombinant laccase could effectively decolour 0.05 mM of MG in 1 h. CcLCC6I could inhibit the toxicity of MG to P. pastoris. This is the first report on the successful expression in P. pastoris of CcLCC6I and its enzymatic property. Laccase can also be considered as a candidate for treating industrial effluent containing MG. PMID:24178808

  15. Disseminated avian mycobacteriosis in a free-living grey heron (Ardea cinerea).

    PubMed

    Quesada-Canales, Oscar; Díaz-Delgado, Josué; Paz, Yania; Domínguez, Lucas; Bezos, Javier; Calabuig, Pascual; Suárez-Bonnet, Alejandro; Fernández, Antonio; Andrada, Marisa

    2013-09-01

    Wild birds share with humans the capacity for moving fast over large distances. During migratory movements, birds carry pathogens that can be transmitted between species. One of these concerning pathogens is Mycobacterium spp. A necropsy was performed in a grey heron (Ardea cinerea) that had been medically treated for a polyarthritic process. Grossly, firm white-yellowish nodules of various size, resembling granulomas, were observed in right carpal joint, both patellar joints, neck musculature, palate, pharynx, larynx, nasal sinuses, pericardial sac, air sacs, proventriculus and intestinal serosa, pancreas, kidneys, adrenal glands, and oviduct. Microscopically, these lesions were composed of multinucleated giant cells, large macrophages, and lymphocytes, with a central zone of necrosis and in some cases with peripheral fibrosis. Acid-fast bacilli were detected within these lesions. Lesions were cultured and Mycobacterium avium subsp. avium was identified. To the best of our knowledge this is the first description of mycobacteriosis in grey herons. Although the grey heron is currently considered a least-concern species, avian mycobacteriosis remains a threat on the conservation management of endangered avian species, and constitutes a public health concern as well. PMID:24283143

  16. Preliminary evidence for color stimuli discrimination in the Asian small-clawed otter (Aonyx cinerea).

    PubMed

    Svoke, Joseph T; Snyder, Rebecca J; Elgart, Jenny Brink

    2014-06-01

    Color discrimination ability can be determined through anatomy or perceptual ability. In this study we tested perceptual ability. Three Asian small-clawed otters (Aonyx cinerea), one male and two females, were trained via operant conditioning to discriminate stimuli within a training task. If they passed criteria for this task, they were tested on as many as six delayed matching-to-sample experimental tasks. These experimental tasks involved comparing varying saturations of the colors blue, green, and red against varying shades of gray, as well as against each other. The male reached criterion on five of the experimental tasks, indicating an ability to discriminate the stimuli. One female participated in only two tasks and did not achieve the criteria as set. The second female did not pass the training task, and thus was not experimentally tested. This study overall showed some early evidence that Asian small-clawed otters may have the ability to learn to discriminate different stimuli on the basis of color cues. Sensory studies conducted on two other otter species and the results of this study indicate that color vision may be a common trait across Lutrinae species. PMID:24788089

  17. Four cleaved amplified polymorphic sequence (CAPS) markers for the detection of the Juglans ailantifolia chloroplast in putatively native J. cinerea populations.

    PubMed

    McCleary, Tim S; Robichaud, Rodney L; Nuanes, Steve; Anagnostakis, Sandra L; Schlarbaum, Scott E; Romero-Severson, Jeanne

    2009-03-01

    Hybridization between butternut (Juglans cinerea), a forest tree native to eastern North America, and Japanese walnut (J. ailantifolia), a tree tolerant to the lethal fungal disease butternut canker, casts doubt on the genetic identity of the remaining butternuts. We report a diagnostic test to distinguish the J. cinerea chloroplast from the J. ailantifolia chloroplast using cleaved amplified polymorphic sequences resolvable in 1.5% agarose gels. J. ailantifolia maternal ancestry in naturally regenerated stands provides a site selection criterion for studies of introgression dynamics when the non-native parent and the hybrids tolerate a disease to which the native species is susceptible. PMID:21564682

  18. Whisker spot patterns: a noninvasive method of individual identification of Australian sea lions (Neophoca cinerea)

    PubMed Central

    Osterrieder, Sylvia K.; Salgado Kent, Chandra; Anderson, Carlos J. R.; Parnum, Iain M.; Robinson, Randall W.

    2015-01-01

    Reliable methods for identification of individual animals are advantageous for ecological studies of population demographics and movement patterns. Photographic identification, based on distinguishable patterns, unique shapes, or scars, is an effective technique already used for many species. We tested whether photographs of whisker spot patterns could be used to discriminate among individual Australian sea lion (Neophoca cinerea). Based on images of 53 sea lions, we simulated 5,000 patterns before calculating the probability of duplication in a study population. A total of 99% (± 1.5 SD) of patterns were considered reliable for a population of 50, 98% (± 1.7 SD) for 100, 92% (± 4.7 SD) for 500, and 88% (± 5.7 SD) for 1,000. We tested a semiautomatic approach by matching 16 known individuals at 3 different angles (70°, 90°, and 110°), 2 distances (1 and 2 m), and 6 separate times over a 1-year period. A point-pattern matching algorithm for pairwise comparisons produced 90% correct matches of photographs taken on the same day at 90°. Images of individuals at 1 and 2 m resulted in 89% correct matches, those photographed at different angles and different times (at 90°) resulted in 48% and 73% correct matches, respectively. Our results show that the Chamfer distance transform can effectively be used for individual identification, but only if there is very little variation in photograph angle. This point-pattern recognition application may also work for other otariid species. PMID:26937048

  19. 3-Hydroxy-2-butanone and the first encounter fight in the male lobster cockroach, Nauphoeta cinerea

    NASA Astrophysics Data System (ADS)

    Kou, Rong; Chen, Shu-Chun; Chen, Ying-Ru; Ho, Hsiao-Yung

    2006-06-01

    Although agonistic behavior in the male lobster cockroach, Nauphoeta cinerea, has been known for more than 40 years, this is the first study to directly collect and quantify the emitted pheromones. In the present study, emitted volatile pheromones were collected from each male pair for 60 min during the first encounter fight and identified and quantified by gas chromatography mass spectrometry. The major compound collected was 3-hydroxy-2-butanone (3H-2B). The strength of the attack by the dominant male was classified into three categories from weakest (C) to strongest (A). Of the 92 pairs, eight (8.7%) showed no agonistic interactions, and the amount of 3H-2B collected was 121.8±37.7 ng/male pair. In the five pairs (5.4%) displaying attack strength C, the amount of 3H-2B collected was 689.6±273.7 ng/male pair, and the attack duration was 9.6±2.4 min. In the 53 pairs (57.6%) showing attack strength B, the corresponding values were 5396.2±449.0 ng/male pair and 22.7±1.6 min, while those for the 26 pairs (28.3%) showing attack strength A were 7910.4±1120.6 ng/male pair and 24.9±2.9 min. For both attack strengths A and B, a linear relationship was found between the amount of 3H-2B collected and attack duration, suggesting that the longer the duration of the attack, the more 3H-2B was emitted. In addition, the rate of 3H-2B emission for attack strength A was significantly higher than that for attack strength B. Using Vaseline-coating, we demonstrated that, in the first encounter fight, the 3H-2B was emitted by the dominant male.

  20. Epidemiology of hookworm (Uncinaria sanguinis) infection in free-ranging Australian sea lion (Neophoca cinerea) pups.

    PubMed

    Marcus, Alan D; Higgins, Damien P; Gray, Rachael

    2014-09-01

    Understanding the fundamental factors influencing the epidemiology of wildlife disease is essential to determining the impact of disease on individual health and population dynamics. The host-pathogen-environment relationship of the endangered Australian sea lion (Neophoca cinerea) and the parasitic hookworm, Uncinaria sanguinis, was investigated in neonatal pups during summer and winter breeding seasons at two biogeographically disparate colonies in South Australia. The endemic occurrence of hookworm infection in Australian sea lion pups at these sites was 100% and post-parturient transmammary transmission is likely the predominant route of hookworm infection for pups. The prepatent period for U. sanguinis in Australian sea lion pups was determined to be 11-14 days and the duration of infection approximately 2-3 months. The mean hookworm infection intensity in pups found dead was 2138 ± 552 (n = 86), but a significant relationship between infection intensity and faecal egg count was not identified; infection intensity in live pups could not be estimated from faecal samples. Fluctuations in infection intensity corresponded to oscillations in the magnitude of colony pup mortality, that is, higher infection intensity was significantly associated with higher colony pup mortality and reduced pup body condition. The dynamic interaction between colony, season, and host behaviour is hypothesised to modulate hookworm infection intensity in this species. This study provides a new perspective to understanding the dynamics of otariid hookworm infection and provides evidence that U. sanguinis is a significant agent of disease in Australian sea lion pups and could play a role in population regulation in this species. PMID:25056940

  1. Asian small-clawed otters (Amblonyx cinerea): resting and swimming metabolic rates.

    PubMed

    Borgwardt, N; Culik, B M

    1999-03-01

    Open-flow oxygen and carbon dioxide respirometry was used in Neumünster Zoo (Germany) to examine the energy requirements of six Asian small-clawed otters (Amblonyx cinerea) at rest and swimming voluntarily under water. Our aim was to compare their energy requirements with those of other warm-blooded species to elucidate scale effects and to test whether the least aquatic of the three otter species differs markedly from these and its larger relatives. While at rest on land (16 degrees C, n = 26), otters (n = 6, mean body mass 3.1 +/- 0.4 kg) had a respiratory quotient of 0.77 and a resting metabolic rate of 5.0 +/- 0.8 Wkg-1(SD). This increased to 9.1 +/- 0.8 Wkg-1 during rest in water (11-15 degrees C, n = 4) and to 17.6 +/- 1.4 Wkg-1 during foraging and feeding activities in a channel (12 degrees C, n = 5). While swimming under water (n = 620 measurements) in an 11-m long channel, otters preferred a speed range between 0.7 ms-1 and 1.2 ms-1. Transport costs were minimal at 1 ms-1 and amounted to 1.47 +/- 0.24 JN-1 m-1 (n = 213). Metabolic rates of small-clawed otters in air were similar to those of larger otter species, and about double those of terrestrial mammals of comparable size. In water, metabolic rates during rest and swimming were larger than those extrapolated from larger otter species and submerged swimming homeotherms. This is attributed to high thermoregulatory costs, and high body drag at low Reynolds numbers. PMID:10227184

  2. Regulation of gas exchange and haemolymph pH in the cockroach Nauphoeta cinerea.

    PubMed

    Matthews, Philip G D; White, Craig R

    2011-09-15

    Ventilatory control of internal CO(2) plays an important role in regulating extracellular acid-base balance in terrestrial animals. While this phenomenon is well understood among vertebrates, the role that respiration plays in the acid-base balance of insects is in need of much further study. To measure changes in insect haemolymph pH, we implanted micro pH optodes into the haemocoel of cockroaches (Nauphoeta cinerea). They were then exposed to normoxic, hypoxic, hyperoxic and hypercapnic atmospheres while their haemolymph pH, VCO(2) and abdominal ventilation frequency were measured simultaneously. Intratracheal O(2) levels were also measured in separate experiments. It was found that cockroaches breathing continuously control their ventilation to defend a haemolymph pH of 7.3, except under conditions where hypoxia (<10% O(2)) induces hyperventilation, or where ambient hypercapnia is in excess of haemolymph (>1% CO(2)). In contrast, intratracheal O(2) levels fluctuated widely, but on average remained above 15% in normoxic (21% O(2)) atmospheres. Decapitation caused the cockroaches to display discontinuous gas exchange cycles (DGCs). The alternating periods of ventilation and apnoea during DGCs caused haemolymph pH to fluctuate by 0.11 units. Exposure to hypoxia caused haemolymph pH to increase and initiated brief bouts of spiracular opening prior to the active ventilation phase. The spontaneous occurrence of DGCs in decapitated cockroaches indicates that central pattern generators in the thoracic and abdominal ganglia generate the periodic gas exchange pattern in the absence of control from the cephalic ganglion. This pattern continues to maintain gas exchange, but with less precision. PMID:21865519

  3. A DNA Damage Checkpoint Pathway Coordinates the Division of Dikaryotic Cells in the Ink Cap Mushroom Coprinopsis cinerea

    PubMed Central

    de Sena-Tomás, Carmen; Navarro-González, Mónica; Kües, Ursula; Pérez-Martín, José

    2013-01-01

    The fungal fruiting body or mushroom is a multicellular structure essential for sexual reproduction. It is composed of dikaryotic cells that contain one haploid nucleus from each mating partner sharing the same cytoplasm without undergoing nuclear fusion. In the mushroom, the pileus bears the hymenium, a layer of cells that includes the specialized basidia in which nuclear fusion, meiosis, and sporulation occur. Coprinopsis cinerea is a well-known model fungus used to study developmental processes associated with the formation of the fruiting body. Here we describe that knocking down the expression of Atr1 and Chk1, two kinases shown to be involved in the response to DNA damage in a number of eukaryotic organisms, dramatically impairs the ability to develop fruiting bodies in C. cinerea, as well as other developmental decisions such as sclerotia formation. These developmental defects correlated with the impairment in silenced strains to sustain an appropriated dikaryotic cell cycle. Dikaryotic cells in which chk1 or atr1 genes were silenced displayed a higher level of asynchronous mitosis and as a consequence aberrant cells carrying an unbalanced dose of nuclei. Since fruiting body initiation is dependent on the balanced mating-type regulator doses present in the dikaryon, we believe that the observed developmental defects were a consequence of the impaired cell cycle in the dikaryon. Our results suggest a connection between the DNA damage response cascade, cell cycle regulation, and developmental processes in this fungus. PMID:23792951

  4. Absence of a gene encoding cytosine deaminase in the genome of the agaricomycete Coprinopsis cinerea enables simple marker recycling through 5-fluorocytosine counterselection.

    PubMed

    Nakazawa, Takehito; Honda, Yoichi

    2015-08-01

    Coprinopsis cinerea is a model species for molecular genetics studies of sexual development in agaricomycetes or homobasidiomycetes. Recently, efficient gene targeting was established in this fungus by generating Cc.ku70 or Cc.lig4 disruptants. To determine the molecular mechanisms underlying sexual development, which involves many genes, generating multiple gene disruptants is required. However, the number of transformation markers available for C. cinerea is limited. This problem would be solved by establishing marker recycling. In this study, we found that C. cinerea lacks a gene encoding a homolog of Saccharomyces cerevisiae cytosine deaminase (Fcy1p) in its genome, which is present in many other fungi. We also observed that C. cinerea is resistant to 5-fluorocytosine. Based on these findings, we established a simple marker recycling method in this fungus using 5-fluorocytosine counterselection after heterologous expression of FCY1 derived from Pleurotus ostreatus, together with the hygromycin resistance gene. This study proposes a simple genetic manipulation system that can be performed using wild-type strains of several fungi that lack a gene homologous to S. cerevisiae FCY1 in their genomes. PMID:26223587

  5. Transglycosylation Activity of Glycosynthase Mutants of Endo-β-N-Acetylglucosaminidase from Coprinopsis cinerea

    PubMed Central

    Eshima, Yasunari; Higuchi, Yujiro; Kinoshita, Takashi; Nakakita, Shin-ichi; Takegawa, Kaoru

    2015-01-01

    Endo-β-N-acetylglucosaminidase (ENGase), which catalyzes hydrolysis of N-linked oligosaccharides, is a useful tool for analyzing oligosaccharide contents of glycoproteins. However, there are only a few known ENGases that can catalyze the hydrolysis of human complex type oligosaccharides, and although commercially available, they are expensive. Here, we report the cloning of two ENGase encoding cDNAs from the basidiomycete fungus Coprinopsis cinerea, Endo-CC1 and Endo-CC2. We successfully expressed recombinant His6-tagged Endo-CC1 and Endo-CC2 in Escherichia coli and purified them for enzymatic characterization. Both Endo-CC1 and Endo-CC2 showed hydrolytic activity on high-mannose and complex type oligosaccharides. Since Endo-CC1 could be prepared more easily than Endo-CC2 from E. coli cultures, we examined the enzymatic properties of Endo-CC1 in detail. Our results showed that Endo-CC1 acted on both N-linked high-mannose type and sialobiantennary type complex oligosaccharides of glycoproteins RNase B and human transferrin, respectively, but not on the sialotriantennary type complex oligosaccharide of glycoprotein fetuin. Examination of the transglycosylation activity of Endo-CC1 revealed that the wild-type Endo-CC1 could not transfer the sialobiantennary type complex oligosaccharide onto the deglycosylated RNase B. To obtain an Endo-CC1 mutant with desired transglycosylation activity, we performed mutation analysis of the active site residue Asn 180 (N180), known to be important for catalysis, by individually replacing it with the remaining 19 amino acid residues. Transglycosylation analyses of these mutants led us to identify one mutant, namely Endo-CC1N180H, which exhibited the desired transglycosylation activity. Taken together, we suggest that Endo-CC1 would potentially be a valuable tool for analyzing oligosaccharides on glycoproteins, as large quantities of it could be made available more easily and less expensively than the currently used enzyme, Endo

  6. Strategic 3-hydroxy-2-butanone release in the dominant male lobster cockroach, Nauphoeta cinerea

    NASA Astrophysics Data System (ADS)

    Chen, Shu-Chun; Yang, Rou-Ling; Ho, Hsiao-Yung; Chou, Szu-Ying; Kou, Rong

    2007-11-01

    In the lobster cockroach Nauphoete cinerea, the dominant subordinate hierarchy formed via the agonistic interactions is unstable, and changes in rank order are common. Our previous results showed that in the first encounter fight during initial rank formation, microgram levels of 3H-2B are released by the aggressive posture (AP)-adopting dominant male. In the present study, the pattern of daily pheromone (3H-2B) release during the domination period and on the day of rank switch, rank duration, and rank switch frequency were investigated in three-male groups and six-male groups to examine the effect of higher frequency of agonistic encounters. The results showed that, in the three-male groups (50-day observation period), daily 3H-2B release rate was not constant, but fluctuated, the average duration of dominant rank was 16.6 ± 2.0 days, rank switch occurred in 58.8% of groups, and the frequency of rank switching (average number of rank switches/group/50 days) was 1.4 ± 0.2. For the six-male groups (30-day observation period), the daily 3H-2B release rate also fluctuated, but the duration of dominant rank was significantly shorter at 4.2 ± 0.6 days, rank switch occurred in 100% of groups, and the frequency of rank switching (average number of rank switches/group/30 days) was significantly higher at 6.9 ± 0.6. The results for both sets of male groups showed that as a new rank formed (either on the first encounter day or on the day of rank switching), the dominant status was significantly associated with a higher 3H-2B release rate. In the animal kingdom, fighting usually involves communication or the exchange of signals, and the results of this study indicated that the fluctuating daily 3H-2B release rate adopted by the dominants is a kind of strategic release and the 3H-2B release rate is a signal used to determine dominance.

  7. Characterization of a Novel Bipartite Double-Stranded RNA Mycovirus Conferring Hypovirulence in the Phytopathogenic Fungus Botrytis porri

    PubMed Central

    Wu, Mingde; Jin, Fengyin; Zhang, Jing; Yang, Long; Jiang, Daohong

    2012-01-01

    The ascomycete Botrytis porri causes clove rot and leaf blight of garlic worldwide. We report here the biological and molecular features of a novel bipartite double-stranded RNA (dsRNA) mycovirus named Botrytis porri RNA virus 1 (BpRV1) from the hypovirulent strain GarlicBc-72 of B. porri. The BpRV1 genome comprises two dsRNAs, dsRNA-1 (6,215 bp) and dsRNA-2 (5,879 bp), which share sequence identities of 62 and 95% at the 3′- and 5′-terminal regions, respectively. Two open reading frames (ORFs), ORF I (dsRNA-1) and ORF II (dsRNA-2), were detected. The protein encoded by the 3′-proximal coding region of ORF I shows sequence identities of 19 to 23% with RNA-dependent RNA polymerases encoded by viruses in the families Totiviridae, Chrysoviridae, and Megabirnaviridae. However, the proteins encoded by the 5′-proximal coding region of ORF I and by the entire ORF II lack sequence similarities to any reported virus proteins. Phylogenetic analysis showed that BpRV1 belongs to a separate clade distinct from those of other known RNA mycoviruses. Purified virions of ∼35 nm in diameter encompass dsRNA-1 and dsRNA-2, and three structural proteins (SPs) of 70, 80, and 85 kDa, respectively. Peptide mass fingerprinting analysis revealed that the 80- and 85-kDa SPs are encoded by ORF I, while the 70-kDa SP is encoded by ORF II. Introducing BpRV1 purified virions into the virulent strain GarlicBc-38 of B. porri caused derivative 38T reduced mycelial growth and hypovirulence. These combined results suggest that BpRV1 is a novel bipartite dsRNA virus that possibly belongs to a new virus family. PMID:22496220

  8. Trophic ecology of the Grey Wagtail Motacilla cinerea before and during the breeding season in the region of Bejaia (Algeria).

    PubMed

    Bougaham, Abdelazize Franck; Moulaï, Riadh; O'Halloran, John

    2014-01-01

    The diet composition of the Grey Wagtail Motacilla cinerea of the Babor Range is followed by analysis of faecal samples (90 faeces) before and during the breeding season 2010. The Grey Wagtail's diet varies depending on the stage of the breeding cycle at the southern edge of their breeding area in North Africa (Bejaia). The diet consists predominantly of aquatic preys (51.79%), with Coleoptera being the most frequent constituent (n=331, 45.5%). During the pre-laying period (February-March), the diet was variable (91 prey-taxa and H'=3,36 bits) and preys of medium size (5 to 8mm) were most common. During the incubation period (April-May), preys were mainly aquatic (60%) and larger (20 to 32mm). At the end of the breeding season (June-July), there was a greater occurrence of terrestrial preys (31 aquatic versus 30 terrestrial taxa). PMID:25103832

  9. Clinical canine parvovirus type 2C infection in a group of Asian small-clawed otters (Aonyx cinerea).

    PubMed

    Gjeltema, Jenessa; Murphy, Hayley; Rivera, Sam

    2015-03-01

    Despite the occurrence of clinical disease in a wide range of carnivore hosts, only vague accounts of clinical canine parvovirus type 2 (CPV-2) in any otter species have been reported in the literature. Over the course of 25 days, nine Asian small-clawed otters (Aonyx cinerea) presented for evaluation of inappetence, lethargy, vomiting, and diarrhea. A diagnosis of canine parvovirus type 2c was made based on electron microscopy, polymerase chain reaction, and DNA sequencing of group fecal samples. Supportive care was provided based on individual clinical assessment and included subcutaneous crystalline fluid therapy, antiemetics, antibiotics, appetite stimulants, and a neuraminidase inhibitor. Five of the nine otters exhibited moderate to severe disease requiring treatment, and one case was fatal despite supportive efforts. In light of this case report, CPV-2 should be recognized as a potential cause of gastrointestinal disease in Asian small-clawed otters. PMID:25831584

  10. EXTRACORPOREAL SHOCK WAVE LITHOTRIPSY AND ENDOSCOPIC URETERAL STENT PLACEMENT IN AN ASIAN SMALL-CLAWED OTTER (AONYX CINEREA) WITH NEPHROLITHIASIS.

    PubMed

    Wojick, Kimberlee B; Berent, Allyson C; Weisse, Chick W; Gamble, Kathryn C

    2015-06-01

    Urolithiasis is a significant disease concern in Asian small-clawed otters (Aonyx cinerea), with over 60% of captive animals affected. Bilateral ureteral stent placement, using endoscopic and fluoroscopic guidance, and extracorporeal shock wave lithotripsy (ESWL) were performed as salvage procedures in a 13-yr-old intact female Asian small-clawed otter following a 7-yr history of nephrolithiasis and progressive renal insufficiency. Following the procedure, radiographs revealed a slight shifting of urolith position, although a decrease in urolith mass was not observed. As a result of declining quality of life related to severe osteoarthritis, the otter was euthanized 5 wk after the procedure. While this treatment approach was unsuccessful in this case, the technique was clinically feasible, so ESWL and ureteral stent placement may remain a consideration for other individuals of this species presented earlier in the course of this disease. PMID:26056891

  11. Lupeol, a pentacyclic triterpenoid isolated from Vernonia cinerea attenuate selenite induced cataract formation in Sprague Dawley rat pups.

    PubMed

    Asha, Radha; Gayathri Devi, V; Abraham, Annie

    2016-02-01

    This study investigated the inhibitory effects of active component isolated from flavonoid fraction of Vernonia cinerea (FVC), lupeol on selenite induced cataract formation. Previous reports suggest that phytochemicals or natural plant products retard the process of cataractogenesis by scavenging free oxygen radicals. Hence, the present study sought to assess the potential of lupeol on in vivo selenite induced cataract models. Lupeol, a pentacyclic triterpenoid, was isolated from the ethyl acetate fraction of methanolic extract of Vernonia cinerea, follows standard chromatographic techniques. Structural elucidation of the compound was carried out using (1)H NMR, (13)C NMR, Mass spectrometry together with other complementary techniques (UV and IR). From these, the isolated compound was identified as Lupeol (3'-hydroxylup-20(29)-ene). The antioxidant activity was comparatively studied using DPPH radical scavenging and FRAP assay. Lupeol exhibited higher DPPH radical scavenging activity as well as reducing power assay. In this study, cataract was induced by a single subcutaneous injection of sodium selenite (4 μg/g body weight) on rat pups. Lupeol was administered orally from 8th day upto 21st day at a concentration 25 μg/g body weight. Cataract was visualized on 16th day with the help of an ophthalmoscope and later on with the naked eye. On the 30th day, rats were euthanized by sodium pentothal injection, lenses were excised and the biochemical parameters such as activity of superoxide dismutase (SOD), catalase (CAT), Glutathione peroxidase (GPx), Glutathione reductase (GR), Glutathione-S-transferase (GST), Ca(2+) ATPase, glutathione content (GSH), reactive oxygen species (ROS), lipid peroxidation products (malondialdehyde) were estimated and found effective in the treatment of cataract by lupeol. PMID:26697995

  12. Direct proof by 13C-nuclear magnetic resonance of semi-purified extract and isolation of ent-Catechin from leaves of Eucalyptus cinerea

    PubMed Central

    Silva, Sayonara Mendes; Abe, Simone Yae; Bueno, Fernanda Giacomini; Lopes, Norberto Peporine; de Mello, João Carlos Palazzo; Nakashima, Tomoe

    2014-01-01

    Background: Eucalyptus cinerea F. Muell. ex Benth. is native to Australia and acclimatized to Southern Brazil. Its aromatic leaves are used for ornamental purposes and have great potential for essential oil production, although reports of its use in folk medicine are few. Objective: This study evaluated the composition of E. cinerea leaves using the solid state 13C-nuclear magnetic resonance (NMR) and isolation of the compound from the semipurified extract (SE). Materials and Methods: The SE of E. cinerea leaves was evaluated in the solid state by 13C-NMR spectrum, and the SE was chromatographed on a Sephadex LH-20 column, followed by high-speed counter-current chromatography to isolate the compound. The SE was analyzed by 13C-NMR and matrix-assisted laser desorption/ionization-time-of-flight spectra. Results: Flavan-3-ol units were present, suggesting the presence of proanthocyanidins as well as a gallic acid unit. The uncommon ent-catechin was isolated. Conclusion: The presence of ent-catechin is reported for the first time in this genus and species. PMID:25210302

  13. Problem of behavioral plasticity in slave-making Amazon-ant Polyergus rufescens Latr. and in its slave-ants Formica fusca L. and Formica cinerea Mayr.

    PubMed

    Dobrzańska, J

    1978-01-01

    Changes in the natural behavior, expressing adaptation to cohabitation in a community, were observed in the slave-making ants P. rufescens and the slave-species F. fusca and F. cinerea. After a few raids, the initially undirected arousal evoked in the slaves by the amazons' raids, begins to acquire attributes appropriate to the situation, but completely different in both studied slave-species. F. cinerea picks up the pupae abandoned on the nest by the slave-making ants and eventually begins to wrench them away from the amazons arriving with prey. F. fusca, whose nests have openings so narrow that it prevents the mass entry of amazons with prey, begin to enlarge those openings shortly after the amazons return. After a certain number of raids, F. fusca begin to enlarge the openings immediately after the departure of the amazons for the slave-raid. The amazons, on their side, adjust soon to the specific behavior of the given slave-species; in nests with F. fusca, they make use of enlarged openings, carrying their prey through them into the nest; when F. cinerea are the slaves, most of the amazons begin to drop the pupae on the nest, and later even surrender the prey to the slaves who meet them. It is supposed that in all three species, under the influence of specific conditions there occurs the process of learning of new forms of behavior. PMID:749551

  14. Valorization of cauliflower (Brassica oleracea L. var. botrytis) by-products as a source of antioxidant phenolics.

    PubMed

    Llorach, Rafael; Espín, Juan Carlos; Tomás-Barberán, Francisco A; Ferreres, Federico

    2003-04-01

    The present study reports the development of two extraction protocols, with potential industrial applicability, to valorize cauliflower (Brassica oleracea L. var. botrytis) byproducts as a source of antioxidant phenolics. In addition, the nonionic polystyrene resin Amberlite XAD-2 was used to obtain purified extracts. The extract yield, phenolic content, phenolic yield, and correlation between the antioxidant activity and the phenolic content were studied. The water and ethanol protocols yield a phenolic content of 33.8 mg/g freeze-dried extract and 62.1 mg/g freeze-dried extract, respectively. This percentage increased considerably when the extracts were purified using Amberlite XAD-2 yielding a phenolic content of 186 mg/g freeze-dried extract (water extract) and 311.1 mg/g freeze-dried extract (ethanol extract). Cauliflower byproduct extracts showed significant free radical scavenging activity (vs both DPPH(*) and ABTS(*)(+) radicals), ferric reducing ability (FRAP assay), and capacity to inhibit lipid peroxidation (ferric thiocyanate assay). In addition, the antioxidant activity was linearly correlated with the phenolics content. The results obtained indicate that the cauliflower byproducts are a cheap source of antioxidant phenolics very interesting from both the industrial point of view and the possible usefulness as ingredients to functionalize foodstuffs. PMID:12670153

  15. In-depth analysis of internal control genes for quantitative real-time PCR in Brassica oleracea var. botrytis.

    PubMed

    Sheng, X G; Zhao, Z Q; Yu, H F; Wang, J S; Zheng, C F; Gu, H H

    2016-01-01

    Quantitative reverse-transcription PCR (qRT-PCR) is a versatile technique for the analysis of gene expression. The selection of stable reference genes is essential for the application of this technique. Cauliflower (Brassica oleracea L. var. botrytis) is a commonly consumed vegetable that is rich in vitamin, calcium, and iron. Thus far, to our knowledge, there have been no reports on the validation of suitable reference genes for the data normalization of qRT-PCR in cauliflower. In the present study, we analyzed 12 candidate housekeeping genes in cauliflower subjected to different abiotic stresses, hormone treatment conditions, and accessions. geNorm and NormFinder algorithms were used to assess the expression stability of these genes. ACT2 and TIP41 were selected as suitable reference genes across all experimental samples in this study. When different accessions were compared, ACT2 and UNK3 were found to be the most suitable reference genes. In the hormone and abiotic stress treatments, ACT2, TIP41, and UNK2 were the most stably expressed. Our study also provided guidelines for selecting the best reference genes under various experimental conditions. PMID:27525844

  16. Transformation ability of fungi isolated from cork and grape to produce 2,4,6-trichloroanisole from 2,4,6-trichlorophenol.

    PubMed

    Maggi, L; Mazzoleni, V; Fumi, M D; Salinas, M R

    2008-03-01

    The ability of eight fungal strains to transform 2,4,6-trichlorophenol (TCP) to 2,4,6-trichloroanisole (TCA) was studied. These fungi were isolated from cork, belonging to the genera Penicillium, Aspergillus, Trichoderma and Chrysonilia, and from grapes Botrytis cinerea. All, except Chrysonilia, produced TCA when grown directly on cork in the presence of TCP, Aspergillus and Botrytis cinerea being the ones with the highest level of production. It is the first time that Botrytis cinerea, a microorganism often present on grapes and in winery environments, has been shown to transform TCP into TCA. This result can partially explain the wine cork taint before being bottled. PMID:18311620

  17. Trichomes and chemical composition of the volatile oil of Trichogonia cinerea (Gardner) R. M. King & H. Rob. (Eupatorieae, Asteraceae).

    PubMed

    Fernandes, Yanne S; Trindade, Luma M P; Rezende, Maria Helena; Paula, José R; Gonçalves, Letícia A

    2016-03-01

    Trichogonia cinerea is endemic to Brazil and occurs in areas of cerrado and campo rupestre. In this study, we characterized the glandular and non-glandular trichomes on the aerial parts of this species, determined the principal events in the development of the former, and identified the main constituents of the volatile oil produced in its aerial organs. Fully expanded leaves, internodes, florets, involucral bracts, and stem apices were used for the characterization of trichomes. Leaves, internodes, florets, and involucral bracts were examined by light microscopy and scanning electron microscopy, whereas stem apices were examined only by light microscopy. Branches in the reproductive phase were used for the extraction and determination of the composition of the volatile oil. The species has three types of glandular trichomes, biseriate vesicular, biseriate pedunculate, and multicellular uniseriate, which secrete volatile oils and phenolic compounds. The major components identified in the volatile oil were 3,5-muuroladiene (39.56%) and butylated hydroxytoluene (13.07%). PMID:26982621

  18. Purification, characterization and function analysis of an extracellular β-glucosidase from elongating stipe cell walls in Coprinopsis cinerea.

    PubMed

    Zhang, Wenming; Kang, Liqin; Yang, Mingmei; Zhou, Yajun; Wang, Jun; Liu, Zhonghua; Yuan, Sheng

    2016-05-01

    A β-glycoside hydrolase was isolated from cell walls material in Coprinopsis cinerea elongating stipes. By analysis of SDS-PAGE, MALDI-TOF/TOF MS and substrate specificity, this enzyme was characterized as an extracellular β-glucosidase which is a trimer consisting of three homosubunits. β-Glucosidase did not degrade β-glucans with modified ends, whereas it hydrolyzed various β-glucans with free ends and related oligosaccharides with β-1,3-, β-1,4- or β-1,6-linkages. Although this β-glucosidase possesses glycosyltransferase activity on laminarioligosaccharides, it did not transfer glucose residues from laminaritriose to β-glucan in stipe cell walls to produce larger β-glucan molecules; instead, it caused a decrease in the molecular size of stipe wall β-glucan by removing glucose. Relatively, the molecular size of wall β-glucans in the elongating apical stipe was less than that found in the non-elongating basal stipes, and this β-glucosidase was more highly expressed in the elongating apical stipe than in non-elongating basal regions. Therefore, we propose that β-glucosidase functions by trimming or cutting the β-glucan side chains on the β-1,3-glucan backbone to prevent them from forming longer branches, keeping the wall plastic to promote diffuse wall growth. PMID:27030727

  19. Standard metabolic rate is associated with gestation duration, but not clutch size, in speckled cockroaches Nauphoeta cinerea.

    PubMed

    Schimpf, Natalie G; Matthews, Philip G D; White, Craig R

    2012-12-15

    Metabolic rate varies significantly between individuals, and these differences persist even when the wide range of biotic and abiotic factors that influence metabolism are accounted for. It is important to understand the life history implications of variation in metabolic rate, but they remain poorly characterised despite a growing body of work examining relationships between metabolism and a range of traits. In the present study we used laboratory-bred families (one sire to three dams) of Nauphoeta cinerea (Olivier) (speckled cockroaches) to examine the relationship between standard metabolic rate (SMR) and reproductive performance (number of offspring and gestation duration). We show that SMR is negatively associated with female gestation duration. Age at mating is negatively associated with gestation duration for females, and mass is negatively associated with the average gestation duration of the females a male was mated with. In addition to the results in the current literature, the results from the present study suggest that the association between metabolism and life history is more complex than simple relationships between metabolism and various fitness traits. Future work should consider longitudinal, ontogenetic as well as selective and quantitative genetic breeding approaches to fully examine the associations between metabolism and fitness. PMID:23259052

  20. Standard metabolic rate is associated with gestation duration, but not clutch size, in speckled cockroaches Nauphoeta cinerea

    PubMed Central

    Schimpf, Natalie G.; Matthews, Philip G. D.; White, Craig R.

    2012-01-01

    Summary Metabolic rate varies significantly between individuals, and these differences persist even when the wide range of biotic and abiotic factors that influence metabolism are accounted for. It is important to understand the life history implications of variation in metabolic rate, but they remain poorly characterised despite a growing body of work examining relationships between metabolism and a range of traits. In the present study we used laboratory-bred families (one sire to three dams) of Nauphoeta cinerea (Olivier) (speckled cockroaches) to examine the relationship between standard metabolic rate (SMR) and reproductive performance (number of offspring and gestation duration). We show that SMR is negatively associated with female gestation duration. Age at mating is negatively associated with gestation duration for females, and mass is negatively associated with the average gestation duration of the females a male was mated with. In addition to the results in the current literature, the results from the present study suggest that the association between metabolism and life history is more complex than simple relationships between metabolism and various fitness traits. Future work should consider longitudinal, ontogenetic as well as selective and quantitative genetic breeding approaches to fully examine the associations between metabolism and fitness. PMID:23259052

  1. Toxicity induced by Prasiola crispa to fruit fly Drosophila melanogaster and cockroach Nauphoeta cinerea: evidence for bioinsecticide action.

    PubMed

    Zemolin, Ana Paula Pegoraro; Cruz, Litiele Cezar; Paula, Mariane Trindade; Pereira, Betina Kappel; Albuquerque, Margelli Pereira; Victoria, Filipe Carvalho; Pereira, Antônio Batista; Posser, Thaís; Franco, Jeferson Luis

    2014-01-01

    The adverse effects of the alga Prasiola crispa extract (PcE) were investigated in a fruit fly (Drosophila melanogaster) and cockroach (Nauphoeta cinerea) model. In flies, toxicity was assessed as mortality and biochemical alterations including acetylcholinesterase (AChE) activity and oxidative stress markers. The cardiotoxic action of PcE was also examined in a model of semi-isolated cockroach heart. The administration of PcE (2 mg/ml) to flies for 24 h resulted in a marked increase in mortality rate (7.6-fold rise compared to control). AChE activity, glutathione (GSH) levels, and hydroperoxide formation remained unchanged. Fly glutathione S-transferase (GST) and catalase (CAT) activity were significantly altered after PcE treatment. Fraction III (ethyl acetate) of PcE was significantly more toxic to flies compared to fractions I (methanol) and II (ethanol). A significant decrease was noted in cockroach semi-isolated heart function. The addition of 5,5'-dithiobis-(2-nitrobenzoic acid (DTNB), an oxidizing agent, concomitant with the extract significantly blocked this effect, suggesting that reduced compounds may be involved in the cardiotoxic action produced by PcE. Our results show for the first time the adverse effects of PcE in two insect models, Drosophila melanogaster and Nauphoetacinerea. The insecticidal properties of PcE may be related to changes in important antioxidant/detoxifying systems, as well as to changes in insect cardiac function. PMID:24555652

  2. Complete Mitochondrial Genome Sequence of Acrida cinerea (Acrididae: Orthoptera) and Comparative Analysis of Mitochondrial Genomes in Orthoptera

    PubMed Central

    Liu, Nian; Huang, Yuan

    2010-01-01

    The complete 15,599-bp mitogenome of Acrida cinerea was determined and compared with that of the other 20 orthopterans. It displays characteristic gene content, genome organization, nucleotide composition, and codon usage found in other Caelifera mitogenomes. Comparison of 21 orthopteran sequences revealed that the tRNAs encoded by the H-strand appear more conserved than those by the L-stand. All tRNAs form the typical clover-leaf structure except trnS (agn), and most of the size variation among tRNAs stemmed from the length variation in the arm and loop of TΨC and the loop of DHU. The derived secondary structure models of the rrnS and rrnL from 21 orthoptera species closely resemble those from other insects on CRW except a considerably enlarged loop of helix 1399 of rrnS in Caelifera, which is a potentially autapomorphy of Caelifera. In the A+T-rich region, tandem repeats are not only conserved in the closely related mitogenome but also share some conserved motifs in the same subfamily. A stem-loop structure, 16 bp or longer, is likely to be involved in replication initiation in Caelifera and Grylloidea. A long T-stretch (>17 bp) with conserved stem-loop structure next to rrnS on the H-strand, bounded by a purine at either end, exists in the three species from Tettigoniidae. PMID:21197069

  3. Assessing stimulus and subject influences on auditory evoked potentials and their relation to peripheral physiology in green treefrogs (Hyla cinerea).

    PubMed

    Buerkle, Nathan P; Schrode, Katrina M; Bee, Mark A

    2014-12-01

    Anurans (frogs and toads) are important models for comparative studies of communication, auditory physiology, and neuroethology, but to date, most of our knowledge comes from in-depth studies of a relatively small number of model species. Using the well-studied green treefrog (Hyla cinerea), this study sought to develop and evaluate the use of auditory evoked potentials (AEPs) as a minimally invasive tool for investigating auditory sensitivity in a larger diversity of anuran species. The goals of the study were to assess the effects of frequency, signal level, sex, and body size on auditory brainstem response (ABR) amplitudes and latencies, characterize gross ABR morphology, and generate an audiogram that could be compared to several previously published audiograms for green treefrogs. Increasing signal level resulted in larger ABR amplitudes and shorter latencies, and these effects were frequency dependent. There was little evidence for an effect of sex or size on ABRs. Analyses consistently distinguished between responses to stimuli in the frequency ranges of the three previously-described populations of afferents that innervate the two auditory end organs in anurans. The overall shape of the audiogram shared prominent features with previously published audiograms. This study highlights the utility of AEPs as a valuable tool for the study of anuran auditory sensitivity. PMID:25151643

  4. Synthesis of Gold Mediated Biocompatible Nanocomposite of Lactone Enriched Fraction from Sahadevi (Vernonia cinerea Lees): An Assessment of Antimalarial Potential.

    PubMed

    Jyotshna; Shanker, Karuna; Khare, Puja; Tiwari, Nimisha; Mohanty, Shilpa; Bawankule, Dnyaneshwar U; Pal, Anirban

    2016-01-01

    Metals reduction into submicro/nano size through bhasma preparations for therapeutic use is well established in ancient traditional system of Indian medicines i.e. Ayurveda. Recently, nanotechnology has drawn the attention of researchers to develeope various size and shape nanoparicles / composite for number of applications.In this article, we report the enrichment of lactone enriched fraction (LEF) by liquid-liquid portioning of Vernonia cinerea metabolic extract and sysnthesis of mediated nano-gold composite (LEF-AuNPs) in single step process. The morphological characteristic based on transmission electron microscope (TEM) image analysis showed that LEF-AuNPs were predominantly nanopolygons and nanobots in shapes ranging from 50-200 nm in size. Abundance of phytochemicals in both LEF and LEF-AuNPs was dissimilar. In LEF, montanol- a diterpenoid, while in LEF-AuNPs, neophytadiene- a phytanes was the major compound. HPLC profile of relatively polar compounds also varied drastically. In-vitro biocompatibility, cytotoxicity [MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) based assay] and storage stabilitiy of LEF-AuNPs were evaluated. The moderate ability of LEF-AuNPs to restrict parasitaemia, extended mean survival time of mice infected with Plasmodium berghei and lack of any evident toxicity provides new opportunities for the safe delivery and applications of such nanocomposites in malaria therapy. PMID:26876516

  5. Assessing stimulus and subject influences on auditory evoked potentials and their relation to peripheral physiology in green treefrogs (Hyla cinerea)

    PubMed Central

    Buerkle, Nathan P.; Schrode, Katrina M.; Bee, Mark A.

    2014-01-01

    Anurans (frogs and toads) are important models for comparative studies of communication, auditory physiology, and neuroethology, but to date, most of our knowledge comes from in-depth studies of a relatively small number of model species. Using the well-studied green treefrog (Hyla cinerea), this study sought to develop and evaluate the use of auditory evoked potentials (AEPs) as a minimally invasive tool for investigating auditory sensitivity in a larger diversity of anuran species. The goals of the study were to assess the effects of frequency, signal level, sex, and body size on auditory brainstem response (ABR) amplitudes and latencies, characterize gross ABR morphology, and generate an audiogram that could be compared to several previously published audiograms for green treefrogs. Increasing signal level resulted in larger ABR amplitudes and shorter latencies, and these effects were frequency dependent. There was little evidence for an effect of sex or size on ABRs. Analyses consistently distinguished between responses to stimuli in the frequency ranges of the three previously-described populations of afferents that innervate the two auditory end organs in anurans. The overall shape of the audiogram shared prominent features with previously published audiograms. This study highlights the utility of AEPs as a valuable tool for the study of anuran auditory sensitivity. PMID:25151643

  6. The effects of call-like masking diminish after nightly exposure to conspecific choruses in green treefrogs (Hyla cinerea).

    PubMed

    Gall, Megan D; Wilczynski, Walter

    2016-05-01

    One of the major difficulties encountered by animals that select mates using acoustic signals is discriminating individual calls from the background noise generated by other conspecifics. Reducing the effects of conspecific masking could improve discrimination of individual calls from background noise. We used auditory evoked potentials to investigate the effects of forward masking on the responses to artificial calls in male and female treefrogs (Hyla cinerea), as well as whether hearing advertisement calls over several nights, as happens in natural frog choruses, could modify the effects of masking. We found that response amplitude decreased with decreasing interstimulus interval when the masker was equal in amplitude to the stimulus. We also found evidence of a priming effect, whereby response amplitude at lower masker amplitudes was greater than when the target stimulus was not preceded by a masker. Finally, we found that the effect of masking was diminished by 10 nights of chorus exposure (i.e. responses were stronger to target stimuli), whereas there was no change in response in the control group. Our results show that hearing dynamic social stimuli, such as frog choruses, can alter the responses of the auditory periphery in a way that could enhance the detection of and response to conspecific acoustic communication signals. PMID:26944493

  7. Effect of the L499M mutation of the ascomycetous Botrytis aclada laccase on redox potential and catalytic properties.

    PubMed

    Osipov, Evgeny; Polyakov, Konstantin; Kittl, Roman; Shleev, Sergey; Dorovatovsky, Pavel; Tikhonova, Tamara; Hann, Stephan; Ludwig, Roland; Popov, Vladimir

    2014-11-01

    Laccases are members of a large family of multicopper oxidases that catalyze the oxidation of a wide range of organic and inorganic substrates accompanied by the reduction of dioxygen to water. These enzymes contain four Cu atoms per molecule organized into three sites: T1, T2 and T3. In all laccases, the T1 copper ion is coordinated by two histidines and one cysteine in the equatorial plane and is covered by the side chains of hydrophobic residues in the axial positions. The redox potential of the T1 copper ion influences the enzymatic reaction and is determined by the nature of the axial ligands and the structure of the second coordination sphere. In this work, the laccase from the ascomycete Botrytis aclada was studied, which contains conserved Ile491 and nonconserved Leu499 residues in the axial positions. The three-dimensional structures of the wild-type enzyme and the L499M mutant were determined by X-ray crystallography at 1.7 Å resolution. Crystals suitable for X-ray analysis could only be grown after deglycosylation. Both structures did not contain the T2 copper ion. The catalytic properties of the enzyme were characterized and the redox potentials of both enzyme forms were determined: E0 = 720 and 580 mV for the wild-type enzyme and the mutant, respectively. Since the structures of the wild-type and mutant forms are very similar, the change in the redox potential can be related to the L499M mutation in the T1 site of the enzyme. PMID:25372682

  8. Effect of the L499M mutation of the ascomycetous Botrytis aclada laccase on redox potential and catalytic properties

    PubMed Central

    Osipov, Evgeny; Polyakov, Konstantin; Kittl, Roman; Shleev, Sergey; Dorovatovsky, Pavel; Tikhonova, Tamara; Hann, Stephan; Ludwig, Roland; Popov, Vladimir

    2014-01-01

    Laccases are members of a large family of multicopper oxidases that catalyze the oxidation of a wide range of organic and inorganic substrates accompanied by the reduction of dioxygen to water. These enzymes contain four Cu atoms per molecule organized into three sites: T1, T2 and T3. In all laccases, the T1 copper ion is coordinated by two histidines and one cysteine in the equatorial plane and is covered by the side chains of hydrophobic residues in the axial positions. The redox potential of the T1 copper ion influences the enzymatic reaction and is determined by the nature of the axial ligands and the structure of the second coordination sphere. In this work, the laccase from the ascomycete Botrytis aclada was studied, which contains conserved Ile491 and nonconserved Leu499 residues in the axial positions. The three-dimensional structures of the wild-type enzyme and the L499M mutant were determined by X-ray crystallography at 1.7 Å resolution. Crystals suitable for X-ray analysis could only be grown after deglycosylation. Both structures did not contain the T2 copper ion. The catalytic properties of the enzyme were characterized and the redox potentials of both enzyme forms were determined: E 0 = 720 and 580 mV for the wild-type enzyme and the mutant, respectively. Since the structures of the wild-type and mutant forms are very similar, the change in the redox potential can be related to the L499M mutation in the T1 site of the enzyme. PMID:25372682

  9. A chloride tolerant laccase from the plant pathogen ascomycete Botrytis aclada expressed at high levels in Pichia pastoris.

    PubMed

    Kittl, Roman; Mueangtoom, Kitti; Gonaus, Christoph; Khazaneh, Shima Tahvilda; Sygmund, Christoph; Haltrich, Dietmar; Ludwig, Roland

    2012-01-20

    Fungal laccases from basidiomycetous fungi are thoroughly investigated in respect of catalytic mechanism and industrial applications, but the number of reported and well characterized ascomycetous laccases is much smaller although they exhibit interesting catalytic properties. We report on a highly chloride tolerant laccase produced by the plant pathogen ascomycete Botrytis aclada, which was recombinantly expressed in Pichia pastoris with an extremely high yield and purified to homogeneity. In a fed-batch fermentation, 495 mg L(-1) of laccase was measured in the medium, which is the highest concentration obtained for a laccase by a yeast expression system. The recombinant B. aclada laccase has a typical molecular mass of 61,565 Da for the amino acid chain. The pI is approximately 2.4, a very low value for a laccase. Glycosyl residues attached to the recombinant protein make up for approximately 27% of the total protein mass. B. aclada laccase exhibits very low K(M) values and high substrate turnover numbers for phenolic and non-phenolic substrates at acidic and near neutral pH. The enzyme's stability increases in the presence of chloride ions and, even more important, its substrate turnover is only weakly inhibited by chloride ions (I(50)=1.4M), which is in sharp contrast to most other described laccases. This high chloride tolerance is mandatory for some applications such as implantable biofuel cells and laccase catalyzed reactions, which suffer from the presence of chloride ions. The high expression yield permits fast and easy production for further basic and applied research. PMID:22178779

  10. Controlling Botrytis elliptica Leaf Blight on Hybrid Lilies through the Application of Convergent Chemical X-ray Irradiation.

    PubMed

    Hong, Sung-Jun; Koo, Tae-Hoon; Yun, Sung-Chul

    2016-04-01

    X-ray irradiation with convergent chemicals such as nano-silver particles or sodium dichloroisocyanurate (NaDCC) has been used to control leaf blight on cut lilies. The oriental hybrid lily cultivars Siberia, Le Reve, and Sorbonne were irradiated five times by 200 Gy of X-rays in 2014. In 2015, Siberia and Sorbonne were irradiated three times by 150 Gy of X-rays. After artificial infection with Botrytis elliptica on the leaves and petals of cut lilies, this study used convergent chemical X-ray irradiation of 200 Gy or 150 Gy. Leaf and petal blight was measured in terms of incidence and severity at 8 days after infection using total 552 cuttings. Results indicate that the treatments of X-ray irradiation and NaDCC in 2014 and 2015 slightly decreased the severity of petal blight on Siberia and Sorbonne. However, the results were not significant and severity did not decrease as NaDCC concentration increased. Vase-life was observed separately after X-ray irradiation of 270 cut lilies in 2014 and 108 cut lilies in 2015. Chlorophyll content was not affected by either 200 Gy or 150 Gy of X-rays. The number of days of fully opened flowers at Siberia of 150 Gy and Le Revu of 200 Gy increased by 1-2 days. In addition, the relative fresh weights of the X-rayed flowers were 10% drier than the non-irradiated controls. Overall, leaf blight control by X-ray was inferior to the control by gamma rays, and petal color was bleached in Sorbonne and Le Reve cvs. by 150 Gy of X-rays. PMID:27147927

  11. Controlling Botrytis elliptica Leaf Blight on Hybrid Lilies through the Application of Convergent Chemical X-ray Irradiation

    PubMed Central

    Hong, Sung-Jun; Koo, Tae-Hoon; Yun, Sung-Chul

    2016-01-01

    X-ray irradiation with convergent chemicals such as nano-silver particles or sodium dichloroisocyanurate (NaDCC) has been used to control leaf blight on cut lilies. The oriental hybrid lily cultivars Siberia, Le Reve, and Sorbonne were irradiated five times by 200 Gy of X-rays in 2014. In 2015, Siberia and Sorbonne were irradiated three times by 150 Gy of X-rays. After artificial infection with Botrytis elliptica on the leaves and petals of cut lilies, this study used convergent chemical X-ray irradiation of 200 Gy or 150 Gy. Leaf and petal blight was measured in terms of incidence and severity at 8 days after infection using total 552 cuttings. Results indicate that the treatments of X-ray irradiation and NaDCC in 2014 and 2015 slightly decreased the severity of petal blight on Siberia and Sorbonne. However, the results were not significant and severity did not decrease as NaDCC concentration increased. Vase-life was observed separately after X-ray irradiation of 270 cut lilies in 2014 and 108 cut lilies in 2015. Chlorophyll content was not affected by either 200 Gy or 150 Gy of X-rays. The number of days of fully opened flowers at Siberia of 150 Gy and Le Revu of 200 Gy increased by 1–2 days. In addition, the relative fresh weights of the X-rayed flowers were 10% drier than the non-irradiated controls. Overall, leaf blight control by X-ray was inferior to the control by gamma rays, and petal color was bleached in Sorbonne and Le Reve cvs. by 150 Gy of X-rays. PMID:27147927

  12. BOTRYTIS-INDUCED KINASE1 Modulates Arabidopsis Resistance to Green Peach Aphids via PHYTOALEXIN DEFICIENT41[W][OPEN

    PubMed Central

    Lei, Jiaxin; A. Finlayson, Scott; Salzman, Ron A.; Shan, Libo; Zhu-Salzman, Keyan

    2014-01-01

    BOTRYTIS-INDUCED KINASE1 (BIK1) plays important roles in induced defense against fungal and bacterial pathogens in Arabidopsis (Arabidopsis thaliana). Its tomato (Solanum lycopersicum) homolog is required for host plant resistance to a chewing insect herbivore. However, it remains unknown whether BIK1 functions in plant defense against aphids, a group of insects with a specialized phloem sap-feeding style. In this study, the potential role of BIK1 was investigated in Arabidopsis infested with the green peach aphid (Myzus persicae). In contrast to the previously reported positive role of intact BIK1 in defense response, loss of BIK1 function adversely impacted aphid settling, feeding, and reproduction. Relative to wild-type plants, bik1 displayed higher aphid-induced hydrogen peroxide accumulation and more severe lesions, resembling a hypersensitive response (HR) against pathogens. These symptoms were limited to the infested leaves. The bik1 mutant showed elevated basal as well as induced salicylic acid and ethylene accumulation. Intriguingly, elevated salicylic acid levels did not contribute to the HR-like symptoms or to the heightened aphid resistance associated with the bik1 mutant. Elevated ethylene levels in bik1 accounted for an initial, short-term repellence. Introducing a loss-of-function mutation in the aphid resistance and senescence-promoting gene PHYTOALEXIN DEFICIENT4 (PAD4) into the bik1 background blocked both aphid resistance and HR-like symptoms, indicating bik1-mediated resistance to aphids is PAD4 dependent. Taken together, Arabidopsis BIK1 confers susceptibility to aphid infestation through its suppression of PAD4 expression. Furthermore, the results underscore the role of reactive oxygen species and cell death in plant defense against phloem sap-feeding insects. PMID:24963070

  13. Cytotoxic activity of acyl phloroglucinols isolated from the leaves of Eucalyptus cinerea F. Muell. ex Benth. cultivated in Egypt

    PubMed Central

    Soliman, Fathy M.; Fathy, Magda M.; Salama, Maha M.; Al-Abd, Ahmed M.; Saber, Fatema R.; El-Halawany, Ali M.

    2014-01-01

    Two acyl phloroglucinol compounds namely; Sideroxylonal B (1) and Macrocarpal A (2) were isolated from the Sideroxylonal-Rich Extract (SRE) of the juvenile leaves of Eucalyptus cinerea; F. Muell. ex Benth cultivated in Egypt. Identification of the isolated compounds was established on the basis of physico-chemical properties and spectral analysis (1D & 2D NMR). The two compounds were isolated for the first time from this species. The SRE alongside with the isolated compounds were tested against three human cancer cell lines; MCF7 (breast carcinoma cell line), HEP2 (laryngeal carcinoma), CaCo (colonic adenocarcinoma) and one type of normal human cell line;10 FS (fibroblast cells). The SRE, (1), and (2) showed cytotoxic activity with IC50 13.6 ± 0.62, 7.2 ± 0.5, 14.8 ± 0.55 μg mL−1 against HEP2 respectively, 11.6 ± 0.47, 4 ± 0.36, 11.4 ± 0.45 μg mL−1 against CaCo, respectively, and 8.6 ± 0.29, 4.4 ± 0.25, and 7.8 ± 0.3 μg mL−1 against MCF7, respectively. Meanwhile, the (SRE) together with (1) and (2) exhibited low cytotoxicity against normal cell line 10 FS, with IC50 55.4 ± 1.4, 43 ± 0.8 and 50.1 ± 1.12 μg mL−1, respectively. The antiprofilerative activity of the tested compounds was evaluated. The cell cycle profile of cells treated with Sideroxylonal-B and Macrocarpal-A indicates possible S-phase specific effects. PMID:24986654

  14. Health assessment of free-ranging endangered Australian sea lion (Neophoca cinerea) pups: effect of haematophagous parasites on haematological parameters.

    PubMed

    Marcus, Alan D; Higgins, Damien P; Gray, Rachael

    2015-06-01

    Evaluation of the health status of free-ranging populations is important for understanding the impact of disease on individuals and on population demography and viability. In this study, haematological reference intervals were developed for free-ranging endangered Australian sea lion (Neophoca cinerea) pups within the context of endemic hookworm (Uncinaria sanguinis) infection and the effects of pathogen, host, and environment factors on the variability of haematological parameters were investigated. Uncinaria sanguinis was identified as an important agent of disease, with infection causing regenerative anaemia, hypoproteinaemia, and a predominantly lymphocytic-eosinophilic systemic inflammatory response. Conversely, the effects of sucking lice (Antarctophthirus microchir) were less apparent and infestation in pups appears unlikely to cause clinical impact. Overall, the effects of U. sanguinis, A. microchir, host factors (standard length, body condition, pup sex, moult status, and presence of lesions), and environment factors (capture-type and year of sampling) accounted for 26-65% of the total variance observed in haematological parameters. Importantly, this study demonstrated that anaemia in neonatal Australian sea lion pups is not solely a benign physiological response to host-environment changes, but largely reflects a significant pathological process. This impact of hookworm infection on pup health has potential implications for the development of foraging and diving behaviour, which would subsequently influence the independent survival of juveniles following weaning. The haematological reference intervals developed in this study can facilitate long-term health surveillance, which is critical for the early recognition of changes in disease impact and to inform conservation management. PMID:25724096

  15. Giardia duodenalis and Cryptosporidium occurrence in Australian sea lions (Neophoca cinerea) exposed to varied levels of human interaction

    PubMed Central

    Delport, Tiffany C.; Asher, Amy J.; Beaumont, Linda J.; Webster, Koa N.; Harcourt, Robert G.; Power, Michelle L.

    2014-01-01

    Giardia and Cryptosporidium are amongst the most common protozoan parasites identified as causing enteric disease in pinnipeds. A number of Giardia assemblages and Cryptosporidium species and genotypes are common in humans and terrestrial mammals and have also been identified in marine mammals. To investigate the occurrence of these parasites in an endangered marine mammal, the Australian sea lion (Neophoca cinerea), genomic DNA was extracted from faecal samples collected from wild populations (n = 271) in Southern and Western Australia and three Australian captive populations (n = 19). These were screened using PCR targeting the 18S rRNA of Giardia and Cryptosporidium. Giardia duodenalis was detected in 28 wild sea lions and in seven captive individuals. Successful sequencing of the 18S rRNA gene assigned 27 Giardia isolates to assemblage B and one to assemblage A, both assemblages commonly found in humans. Subsequent screening at the gdh and β-giardin loci resulted in amplification of only one of the 35 18S rRNA positive samples at the β-giardin locus. Sequencing at the β-giardin locus assigned the assemblage B 18S rRNA confirmed isolate to assemblage AI. The geographic distribution of sea lion populations sampled in relation to human settlements indicated that Giardia presence in sea lions was highest in populations less than 25 km from humans. Cryptosporidium was not detected by PCR screening in either wild colonies or captive sea lion populations. These data suggest that the presence of G. duodenalis in the endangered Australian sea lion is likely the result of dispersal from human sources. Multilocus molecular analyses are essential for the determination of G. duodenalis assemblages and subsequent inferences on transmission routes to endangered marine mammal populations. PMID:25426423

  16. MOLECULAR DETECTION OF ANTIBIOTIC-RESISTANCE DETERMINANTS IN ESCHERICHIA COLI ISOLATED FROM THE ENDANGERED AUSTRALIAN SEA LION (NEOPHOCA CINEREA).

    PubMed

    Delport, Tiffany C; Harcourt, Robert G; Beaumont, Linda J; Webster, Koa N; Power, Michelle L

    2015-07-01

    Greater interaction between humans and wildlife populations poses significant risks of anthropogenic impact to natural ecosystems, especially in the marine environment. Understanding the spread of microorganisms at the marine interface is therefore important if we are to mitigate adverse effects on marine wildlife. We investigated the establishment of Escherichia coli in the endangered Australian sea lion (Neophoca cinerea) by comparing fecal isolation from wild and captive sea lion populations. Fecal samples were collected from wild colonies March 2009-September 2010 and from captive individuals March 2011-May 2013. Using molecular screening, we assigned a phylotype to E. coli isolates and determined the presence of integrons, mobile genetic elements that capture gene cassettes conferring resistance to antimicrobial agents common in fecal coliforms. Group B2 was the most abundant phylotype in all E. coli isolates (n = 37), with groups A, B1, and D also identified. Integrons were not observed in E. coli (n = 21) isolated from wild sea lions, but were identified in E. coli from captive animals (n = 16), from which class I integrases were detected in eight isolates. Sequencing of gene cassette arrays identified genes conferring resistance to streptomycin-spectinomycin (aadA1) and trimethoprim (dfrA17, dfrB4). Class II integrases were not detected in the E. coli isolates. The frequent detection in captive sea lions of E. coli with resistance genes commonly identified in human clinical cases suggests that conditions experienced in captivity may contribute to establishment. Identification of antibiotic resistance in the microbiota of Australian sea lions provides crucial information for disease management. Our data will inform conservation management strategies and provide a mechanism to monitor microorganism dissemination to sensitive pinniped populations. PMID:25919463

  17. Sensitivity of mechanically transmitted pathogens to different disinfectants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The intensive hands-on activities of greenhouse tomato propagation and production favor the spread of mechanically transmitted pathogens, particularly Clavibacter michiganensis subsp. michiganensis (Cmm), viruses, viroids, and Botrytis cinerea. These pathogens can spread during crop handling, graft...

  18. Antifungal activity of the stem bark of Ailanthus excelsa.

    PubMed

    Joshi, B C; Pandey, Anuj; Chaurasia, Leena; Pal, Mahesh; Sharma, R P; Khare, Anakshi

    2003-12-01

    The methanol extract of stem barks of Alianthus excelsa was partitioned with chloroform. The chloroform extract showed fungistatic and fungicidal activity against Aspergillus niger, A. fumigatus, Penicillium frequentence, P. notatum and Botrytis cinerea. PMID:14630175

  19. Evaluation of the toxicity of Streptomyces aburaviensis (R9) towards various agricultural pests

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The culture filtrate fraction extracted with dichloromethane from Streptomyces aburaviensis -R9 strain grown on glucose-peptone-molasses (GPM) broth was bioassayed for its effect on phytopathogenic fungi (Colletotrichum acutatum, C. fragariae, C. gloeosoprioids, Botrytis cinerea, Fusarium oxysporum,...

  20. In planta production of the highly potent resveratrol analogue pterostilbene via stilbene synthase and O-methyltransferase co-expression

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Stilbenoid phytoalexins such as resveratrol, play an important defense role in several plant species against pathogens such as Botrytis cinerea. In addition to their antimicrobial properties, resveratrol and related stilbenes have also generated considerable interest as nutraceuticals, due to their...

  1. Efficient gene targeting in ΔCc.ku70 or ΔCc.lig4 mutants of the agaricomycete Coprinopsis cinerea.

    PubMed

    Nakazawa, Takehito; Ando, Yuki; Kitaaki, Kohei; Nakahori, Kiyoshi; Kamada, Takashi

    2011-10-01

    Coprinopsis cinerea is a model for studies of sexual development in agaricomycetes (homobasidiomycetes). Efficient gene targeting should facilitate such studies, especially because increasing genome and transcriptome information is now available in C. cinerea. To estimate the frequency of gene disruption by homologous integration in this fungus, we tried to disrupt Cc.wc-2, which encodes a WC-2 homolog, a partner of the fungal blue-light photoreceptor, WC-1. Disruption of Cc.wc-2 did not occur when recipients (protoplasts) of the disrupting construct were prepared from asexual spores, oidia, from the wild type, 326, while it occurred when protoplasts were prepared from mycelial cells from the same strain, albeit at a low frequency (3%). Double-stranded RNA-mediated silencing of a ku70 homolog, named Cc.ku70, or the lig4 homolog Cc.lig4 more or less increased the frequency of Cc.wc-2 targeting. On the basis of these results, we disrupted Cc.ku70 using a Cc.lig4-silenced strain. We then disrupted Cc.lig4 using the Cc.ku70 disruptant. We found that the disruption of Cc.ku70 or Cc.lig4 greatly enhanced gene targeting. In addition, this study demonstrates that Cc.wc-2 is involved in blue light perception in this fungus. PMID:21704178

  2. Effect of the L499M mutation of the ascomycetous Botrytis aclada laccase on redox potential and catalytic properties

    SciTech Connect

    Osipov, Evgeny; Kittl, Roman; Shleev, Sergey; Dorovatovsky, Pavel; Tikhonova, Tamara; Popov, Vladimir

    2014-11-01

    The structures of the ascomycetous B. aclada laccase and its L499M T1-site mutant have been solved at 1.7 Å resolution. The mutant enzyme shows a 140 mV lower redox potential of the type 1 copper and altered kinetic behaviour. The wild type and the mutant have very similar structures, which makes it possible to relate the changes in the redox potential to the L499M mutation Laccases are members of a large family of multicopper oxidases that catalyze the oxidation of a wide range of organic and inorganic substrates accompanied by the reduction of dioxygen to water. These enzymes contain four Cu atoms per molecule organized into three sites: T1, T2 and T3. In all laccases, the T1 copper ion is coordinated by two histidines and one cysteine in the equatorial plane and is covered by the side chains of hydrophobic residues in the axial positions. The redox potential of the T1 copper ion influences the enzymatic reaction and is determined by the nature of the axial ligands and the structure of the second coordination sphere. In this work, the laccase from the ascomycete Botrytis aclada was studied, which contains conserved Ile491 and nonconserved Leu499 residues in the axial positions. The three-dimensional structures of the wild-type enzyme and the L499M mutant were determined by X-ray crystallography at 1.7 Å resolution. Crystals suitable for X-ray analysis could only be grown after deglycosylation. Both structures did not contain the T2 copper ion. The catalytic properties of the enzyme were characterized and the redox potentials of both enzyme forms were determined: E{sub 0} = 720 and 580 mV for the wild-type enzyme and the mutant, respectively. Since the structures of the wild-type and mutant forms are very similar, the change in the redox potential can be related to the L499M mutation in the T1 site of the enzyme.

  3. Altered growth and polyamine catabolism following exposure of the chocolate spot pathogen Botrytis fabae to the essential oil of Ocimum basilicum.

    PubMed

    Oxenham, Senga K; Svoboda, Katja P; Walters, Dale R

    2005-01-01

    Biomass of the fungal pathogen Botrytis fabae in liquid culture amended with two chemotypes of the essential oil of basil, Ocimum basilicum, was reduced significantly at concentrations of 50 ppm or less. The methyl chavicol chemotype oil increased the activity of the polyamine biosynthetic enzyme S-adenosylmethionine decarboxylase (AdoMetDC), but polyamine concentrations were not significantly altered. In contrast, the linalol chemotype oil decreased AdoMetDC activity in B. fabae, although again polyamine concentrations were not altered significantly. However activities of the polyamine catabolic enzymes diamine oxidase (DAO) and polyamine oxidase (PAO) were increased significantly in B. fabae grown in the presence of the essential oil of the two chemotypes. It is suggested that the elevated activities of DAO and PAO may be responsible, in part, for the antifungal effects of the basil oil, possibly via the generation of hydrogen peroxide and the subsequent triggering of programmed cell death. PMID:16392245

  4. Discovery of Lobesia botrana (Denis & Schiffermuller) in California: an invasive species new to North America (Lepidoptera: Tortricidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The European grape berry moth, Lobesia botrana (Denis and Schiffermüller), is one of the most destructive pests of grapes in the Palearctic Region. Larvae feed on the fruit, causing direct damage and promoting secondary infection by Botrytis cinerea Persoon (botrytis bunch rot or gray mold). On 15 S...

  5. Ecofriendly hot water treatment reduces postharvest decay and elicits defense response in kiwifruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hot water treatment (HWT) of fruit is an effective approach for managing postharvest decay of fruits and vegetables. In the present study, the effects of HWT (45 degrees C for 10 min) on the growth of Botrytis cinerea and Penicillium expansum in vitro, and gray (B. cinerea) and blue mold (P. expans...

  6. Improving disease resistance of butternut (Juglans cinerea), a threatened fine hardwood: a case for single-tree selection through genetic improvement and deployment.

    PubMed

    Michler, Charles H; Pijut, Paula M; Jacobs, Douglass F; Meilan, Richard; Woeste, Keith E; Ostry, Michael E

    2006-01-01

    Approaches for the development of disease-resistant butternut (Juglans cinerea L.) are reviewed. Butternut is a threatened fine hardwood throughout its natural range in eastern North America because of the invasion of the exotic fungus, Sirococcus clavigignenti-juglandacearum Nair, Kostichka and Kuntz, which causes butternut canker. Early efforts were made to identify and collect putatively resistant germ plasm, identify vectors and to characterize the disease. More recently, molecular techniques have been employed to genetically characterize both the pathogen and the resistant germ plasm. Much of the host resistance may originate from hybridization with a close Asian relative, Japanese walnut (Juglans ailanthifolia Carr.), and from a few natural phenotypic variants. Further genetic characterization is needed before classical breeding or genetic modification can be used to produce canker-resistant trees. PMID:16203722

  7. Production and characterization of interspecific somatic hybrids between Brassica oleracea var. botrytis and B. nigra and their progenies for the selection of advanced pre-breeding materials.

    PubMed

    Wang, Gui-xiang; Tang, Yu; Yan, Hong; Sheng, Xiao-guang; Hao, Wei-Wei; Zhang, Li; Lu, Kun; Liu, Fan

    2011-10-01

    Somatic hybridization is a potential method for gene transfer from wild relatives to cultivated crops that can overcome sexual incompatibilities of two distantly related species. In this study, interspecific asymmetric somatic hybrids of Brassica oleracea var. botrytis (cauliflower) and Brassica nigra (black mustard) were obtained by protoplast fusion and their backcrossed (BC(3)) and selfed (S(3)) offspring were analyzed. Cytological analysis showed that the B. nigra chromosomes were successively eliminated in the backcrosses with cauliflower. The fertility of the hybrid progenies was quite different due to the asynchronous and abnormal chromosome behavior of pollen mother cells (PMC) during meiosis. Analysis of sequence-related amplified polymorphism (SRAP) showed that all of these hybrids mainly had the DNA banding pattern from the two parents with some alterations. Genetically, the selfed generations were closer to B. nigra, while the backcrossed generations were closer to the cauliflower parent. Analysis of cleaved amplified polymorphic sequences (CAPS) and restriction fragment length polymorphisms (RFLP) showed that all somatic hybrids in this study contained chloroplast (cp) DNA of the donor parent black mustard, while mitochondrial (mt) DNA showed evidence of recombination and variations in the regions analyzed. Furthermore, three BC(3) plants (originated from somatic hybrids 3, 4, 10) with 2-8 B. nigra-derived chromosomes shown by genomic in situ hybridization (GISH) displayed a more cauliflower-like morphology and high resistance to black-rot. These plants were obtained as bridge materials for further analysis and breeding. PMID:21603996

  8. Comparative Analysis of Defense Responses in Chocolate Spot-Resistant and -Susceptible Faba Bean (Vicia faba) Cultivars Following Infection by the Necrotrophic Fungus Botrytis fabae

    PubMed Central

    El-Komy, Mahmoud H.

    2014-01-01

    In this study, resistance responses were investigated during the interaction of Botrytis fabae with two faba bean cultivars expressing different levels of resistance against this pathogen, Nubaria (resistant) and Giza 40 (susceptible). Disease severity was assessed on leaves using a rating scale from 1 to 9. Accumulation levels of reactive oxygen species (ROS), lipid peroxidation and antioxidant enzymes (superoxide dismutase, catalase and ascorbate peroxidase) were measured in leaf tissues at different times of infection. The expression profiles of two pathogenesis-related proteins (PRPs) encoded by the genes PR-1 and β-1,3-glucanase were also investigated using reverse transcription RT-PCR analysis. The accumulation of these defense responses was induced significantly in both cultivars upon infection with B. fabae compared with un-inoculated controls. The resistant cultivar showed weaker necrotic symptom expression, less ROS accumulation, a lower rate of lipid peroxidation and higher activity of the enzymatic ROS scavenging system compared with susceptible cultivar. Interestingly, ROS accumulated rapidly in the resistant leaf tissues and peaked during the early stages of infection, whereas accumulation was stronger and more intense in the susceptible tissues in later stages. Moreover, the response of the resistant cultivar to infection was earlier and stronger, exhibiting high transcript accumulation of the PR genes. These results indicated that the induction of oxidant/antioxidant responses and the accumulation of PRPs are part of the faba bean defense mechanism against the necrotrophic fungus B. fabae with a different intensity and timing of induction, depending on the resistance levels. PMID:25506300

  9. Range-wide distribution of genetic diversity in the North American tree Juglans cinerea: a product of range shifts, not ecological marginality or recent population decline.

    PubMed

    Hoban, Sean M; Borkowski, Daniel S; Brosi, Sunshine L; McCleary, Tim S; Thompson, Laura M; McLachlan, Jason S; Pereira, Marie A; Schlarbaum, Scott E; Romero-Severson, Jeanne

    2010-11-01

    The spatial distribution of genetic diversity is a product of recent and historical ecological processes, as well as anthropogenic activities. A current challenge in population and conservation genetics is to disentangle the relative effects of these processes, as a first step in predicting population response to future environmental change. In this investigation, we compare the influence of contemporary population decline, contemporary ecological marginality and postglacial range shifts. Using classical model comparison procedures and Bayesian methods, we have identified postglacial range shift as the clear determinant of genetic diversity, differentiation and bottlenecks in 29 populations of butternut, Juglans cinerea L., a North American outcrossing forest tree. Although butternut has experienced dramatic 20th century decline because of an introduced fungal pathogen, our analysis indicates that recent population decline has had less genetic impact than postglacial recolonization history. Location within the range edge vs. the range core also failed to account for the observed patterns of diversity and differentiation. Our results suggest that the genetic impact of large-scale recent population losses in forest trees should be considered in the light of Pleistocene-era large-scale range shifts that may have had long-term genetic consequences. The data also suggest that the population dynamics and life history of wind-pollinated forest trees may provide a buffer against steep population declines of short duration, a result having important implications for habitat management efforts, ex situ conservation sampling and population viability analysis. PMID:21040046

  10. The antidepressants venlafaxine ("Effexor") and fluoxetine ("Prozac") produce different effects on locomotion in two species of marine snail, the oyster drill (Urosalpinx cinerea) and the starsnail (Lithopoma americanum).

    PubMed

    Fong, Peter P; Bury, Taylor B; Dworkin-Brodsky, Abigail D; Jasion, Christina M; Kell, Rose C

    2015-02-01

    Human antidepressants have been previously shown to induce foot detachment from the substrate in aquatic snails. Prior to foot detachment, antidepressants also affect snail crawling speed. We tested two commonly prescribed antidepressants, venlafaxine ("Effexor") and fluoxetine ("Prozac") on crawling speed and time to reach the air-water interface in two species of marine snail, the oyster drill Urosalpinx cinerea and the American starsnail Lithopoma americanum. Exposure to venlafaxine increased crawling speed in both species, while fluoxetine slowed them down. Our lowest LOEC (lowest observed effect concentration) was 31.3 μg/L venlafaxine in Urosalpinx. Similarly, snails (L. americanum) exposed to venlafaxine tended to move faster and more often to the air-water interface, but exposure to fluoxetine slowed them down. Our lowest LOEC was 345 μg/L fluoxetine in Lithopoma. These results indicate that venlafaxine boosts locomotion, while fluoxetine reduces it, and both behaviors are preludes to foot detachment. The different effects of these two antidepressants on snail locomotion suggest differing physiological mechanisms of action in marine snails as well as possible ecological consequences. PMID:25481651

  11. Essential Oils from Different Plant Parts of Eucalyptus cinerea F. Muell. ex Benth. (Myrtaceae) as a Source of 1,8-Cineole and Their Bioactivities

    PubMed Central

    Silva, Sayonara Mendes; Abe, Simone Yae; Murakami, Fábio Seigi; Frensch, Gustavo; Marques, Francisco A.; Nakashima, Tomoe

    2011-01-01

    Eucalyptus cinerea, known as silver dollar tree, has few descriptions in traditional medicine. Chemical composition and antimicrobial properties of the essential oils of leaves, flowers and fruits, collected seasonally, were determined by GC/MS and disk diffusion/MIC, respectively. 1,8-Cineole was the main compound, particularly in fresh leaves—Spring (74.98%), dried leaves—Spring (85.32%), flowers—Winter (78.76%) and fruits—Winter (80.97%). Other compounds were found in the aerial parts in all seasons: α-pinene (2.41% to 10.13%), limonene (1.46% to 4.43%), α-terpineol (1.73% to 11.72%), and α-terpinyl acetate (3.04% to 20.44%). The essential oils showed antimicrobial activities against bacteria and yeasts, with the best results being found for the dried autumn and winter leaves oils (MIC < 0.39 mg/mL) against Streptococcus pyogenes. For the other tested microorganisms the following MIC results were found: Staphylococcus aureus— Dried leaves oil from summer (0.78 mg/mL), Pseudomonas aeruginosa—Flowers oil from autumn and fruits oil from winter (1.56 mg/mL) and Candida albicans—Flowers oil from autumn and fruits oils from winter and spring (0.78 mg/mL). PMID:26791641

  12. Heavy metal concentrations in diet and livers of Black-crowned Night Heron Nycticorax nycticorax and Grey Heron Ardea cinerea chicks from Pyeongtaek, Korea.

    PubMed

    Kim, Jungsoo; Koo, Tae-Hoe

    2007-07-01

    This study presents concentrations of iron, manganese, zinc, copper, lead and cadmium in diet and livers of Black-crowned Night Heron Nycticorax nycticorax and Grey Heron Ardea cinerea chicks from Pyeongtaek, Gyeonggi-do, Korea. Heavy metal concentrations of heron chicks were not related to concentrations in the diet. Copper concentrations were significantly greater in the diet of Black-crowned Night Herons (geometric mean = 13.6 wet microg/g) than Grey Herons (7.45 wet microg/g), other metal concentrations did not differ between the diet of two species. Manganese (respectively 3.20 wet microg/g, 1.41 wet microg/g) and cadmium (respectively 13.4 wet microg/kg, 1.41 wet microg/kg) concentrations were higher in livers of Black-crowned Night Heron chicks than Grey Heron chicks, but zinc, iron, copper and lead concentrations in livers did not differ in between two herons. The essential elements were at background levels, however copper concentrations were relatively higher than previously reported from Korea. Lead and cadmium concentrations were within background levels for herons. PMID:17410427

  13. Trace element residues in eggshells of grey heron (Ardea cinerea) and black-crowned night heron (Nycticorax nycticorax) from Nallihan Bird Paradise, Ankara-Turkey.

    PubMed

    Ayaş, Zafer

    2007-05-01

    Concentrations of four trace elements (Cd, Ni, Cu and Pb) were determined in eggshells of grey heron (Ardea cinerea) and black-crowned night herons (Nycticorax nycticorax) from Nallihan Bird Paradise, which is located in the northern part of Sariyar Dam Reservoir, Turkey. Results indicated that, within the same area, these ardeid species differed in the levels of Cd, Cu and Pb in their eggshells (generally grey heron > night heron), possibly because females may have foraged in different habitats and regions. Geometric means found for Cd, Ni, Cu and Pb in eggshells were 0.931 mg/kg, 0.405 mg/kg, 6.755 mg/kg and 4.567 mg/kg, respectively, for grey heron; and 0.230 mg/kg, 0.220 mg/kg, 1.369 mg/kg and 1.108 mg/kg, respectively, for night heron. High bioaccumulation from sediments to eggshells occured for Cu and Pb, while Cd and Ni exhibited low accumulation. Bioaccumulation ratios were calculated as 19.63 (Cu) and 22.9 (Pb) in eggshells of grey herons. In conclusion, eggshells of grey herons appeared to be good bioindicators for monitoring of Cu and Pb in Nallihan Bird Paradise. PMID:17364239

  14. Genome-wide identification and characterization of miRNAs in the hypocotyl and cotyledon of cauliflower (Brassica oleracea L. var. botrytis) seedlings.

    PubMed

    Geng, Meijuan; Li, Hui; Jin, Chuan; Liu, Qian; Chen, Chengbin; Song, Wenqin; Wang, Chunguo

    2014-02-01

    MicroRNAs (miRNAs) are a class of small endogenous, non-coding RNAs that have key regulatory functions in plant growth, development, and other biological processes. Hypocotyl and cotyledon are the two major tissues of cauliflower (Brassica oleracea L. var. botrytis) seedlings. Tissue culture experiments have indicated that the regenerative abilities of these two tissues are significantly different. However, the characterization of miRNAs and their roles in regulating organ development in cauliflower remain unexplored. In the present study, two small RNA libraries were sequenced by Solexa sequencing technology. 99 known miRNAs belonging to 28 miRNA families were identified, in which 6 miRNA families were detected only in Brassicaceae. A total of 162 new miRNA sequences with single nucleotide substitutions corresponding to the known miRNAs, and 32 potentially novel miRNAs were also first discovered. Comparative analysis indicated that 42 of 99 known miRNAs and 17 of 32 novel miRNAs exhibited significantly differential expression between hypocotyl and cotyledon, and the differential expression of several miRNAs was further validated by stem-loop RT-PCR. In addition, 235 targets for 89 known miRNAs and 198 targets for 24 novel miRNAs were predicted, and their functions were further discussed. The expression patterns of several representative targets were also confirmed by qRT-PCR analysis. The results identified that the transcriptional expression patterns of miRNAs were negatively correlated with their targets. These findings gave new insights into the characteristics of miRNAs in cauliflower, and provided important clues to elucidate the roles of miRNAs in the tissue differentiation and development of cauliflower. PMID:24170336

  15. Effect of doe milking and supplementation using Dichrostachys cinerea pods on kid and doe performance in grazing goats during the dry season.

    PubMed

    Maphosa, V; Sikosana, J L N; Muchenje, V

    2009-04-01

    Effects of supplementation and milking of Matebele does four weeks pre-partum to weaning, on milk yield, kid birth weight and growth rate were studied. Forty does were divided into four treatments: supplemented and milked (SM), supplemented and not milked (SNM), not supplemented and milked (NSM); and not supplemented and not milked (NSNM). Supplemented goats were given 200 g/doe/day of Dichrostachys cinerea pods containing 19% CP. The study was carried out in September during time of feed unavailability and animals were grazed on rangelands for seven hours before being given the supplement. Fifty-six kids were born from all the experimental animals and 24 were singles whereas 32 were twins. Milk yield was higher (P < 0.05) in supplemented than non-supplemented does. Although supplementation had no effect on kid birth weights, growth rates were higher (P < 0.05) in the SNM group (103 +/- 10.1 g/day) than in the SM group (74 +/- 10.8 g/day). Among the non-supplemented groups, kids from milked does had higher (P < 0.05) daily gains (85 +/- 10.9 g/day) than those from non-milked does (67 +/- 10.7 g/day). Weaning weights were higher (P < 0.05) in SNM does (12.8 +/- 1.30 g/day) than in the SM does (10.2 +/- 1.17 g/day). Among kids from the non-supplemented does, those from milked does had superior (P < 0.05) weaning weights (11.2 +/- 1.23 g/day) than those from non - milked does (9.2 +/- 1.27 g/day). Mortalities were also lowest in kids born from SNM does. It is concluded that pre-partum to weaning supplementation generally increases milk production which then increases kids growth rates and weaning weights, as well as reduce kid mortalities. PMID:18726706

  16. Ivermectin treatment of free-ranging endangered Australian sea lion (Neophoca cinerea) pups: effect on hookworm and lice infection status, haematological parameters, growth, and survival.

    PubMed

    Marcus, Alan D; Higgins, Damien P; Gray, Rachael

    2015-07-01

    A placebo-controlled study was used to investigate the effectiveness of ivermectin to treat hookworm (Uncinaria sanguinis) and lice (Antarctophthirus microchir) infections in free-ranging Australian sea lion (Neophoca cinerea) pups and to test the hypotheses that these parasitic infections cause anaemia, systemic inflammatory responses, and reduced growth, and contribute towards decreased pup survival. Ivermectin was identified as an effective and safe anthelmintic in this species. Pups administered ivermectin had significantly higher erythrocyte counts and significantly lower eosinophil counts compared to controls at 1-2 months post-treatment, confirming that U. sanguinis and/or A. microchir are causatively associated with disease and demonstrating the positive effect of ivermectin treatment on clinical health parameters. Higher growth rates were not seen in ivermectin-treated pups and, unexpectedly, relatively older pups treated with ivermectin demonstrated significantly reduced growth rates when compared to matched saline-control pups. Differences in survival were not identified between treatment groups; however, this was attributed to the unexpectedly low mortality rate of recruited pups, likely due to the unintended recruitment bias towards pups >1-2 months of age for which mortality due to hookworm infection is less likely. This finding highlights the logistical and practical challenges associated with treating pups of this species shortly after birth at a remote colony. This study informs the assessment of the use of anthelmintics as a tool for the conservation management of free-ranging wildlife and outlines essential steps to further the development of strategies to ensure the effective conservation of the Australian sea lion and its parasitic fauna. PMID:25911163

  17. Heterologous production of cellobiose dehydrogenases from the basidiomycete Coprinopsis cinerea and the ascomycete Podospora anserina and their effect on saccharification of wheat straw.

    PubMed

    Turbe-Doan, Annick; Arfi, Yonathan; Record, Eric; Estrada-Alvarado, Isabel; Levasseur, Anthony

    2013-06-01

    Cellobiose dehydrogenases (CDHs) are extracellular glycosylated haemoflavoenzymes produced by many different wood-degrading and phytopathogenic fungi. Putative cellobiose dehydrogenase genes are recurrently discovered by genome sequencing projects in various phylogenetically distinct fungi. The genomes from the basidiomycete Coprinopsis cinerea and the ascomycete Podospora anserina were screened for candidate cdh genes, and one and three putative gene models were evidenced, respectively. Two putative cdh genes were selected and successfully expressed for the first time in Aspergillus niger. CDH activity was measured for both constructions (CDHcc and CDHpa), and both recombinant CDHs were purified to homogeneity and subsequently characterised. Kinetic constants were determined for several carbohydrates including β-1,4-linked di- and oligosaccharides. Optimal temperature and pH were 60 °C and 5 for CDHcc and 65-70 °C and 6 for CDHpa. Both CDHs showed a broad range of pH stability between 4 and 8. The effect of both CDHs on saccharification of micronized wheat straw by an industrial Trichoderma reesei secretome was determined. The addition of each CDH systematically decreased the release of total reducing sugars, but to different extents and according to the CDH concentration. Analytical methods were carried out to quantify the release of glucose, xylose and gluconic acid. An increase of glucose and xylose was measured at a low CDHcc concentration. At moderated and high CDHcc and CDHpa concentrations, glucose was severely reduced with a concomitant increase of gluconic acid. In conclusion, these results give new insights into the physical and chemical parameters and diversity of basidiomycetous and ascomycetous CDHs. These findings also demonstrated that CDH drastically influenced the saccharification on a natural substrate, and thus, CDH origin, concentration and potential enzymatic partners should be carefully considered in future artificial secretomes for

  18. Characterization of a novel PQQ-dependent quinohemoprotein pyranose dehydrogenase from Coprinopsis cinerea classified into auxiliary activities family 12 in carbohydrate-active enzymes.

    PubMed

    Takeda, Kouta; Matsumura, Hirotoshi; Ishida, Takuya; Samejima, Masahiro; Ohno, Hiroyuki; Yoshida, Makoto; Igarashi, Kiyohiko; Nakamura, Nobuhumi

    2015-01-01

    The basidiomycete Coprinopsis cinerea contains a quinohemoprotein (CcPDH named as CcSDH in our previous paper), which is a new type of pyrroloquinoline-quinone (PQQ)-dependent pyranose dehydrogenase and is the first found among all eukaryotes. This enzyme has a three-domain structure consisting of an N-terminal heme b containing a cytochrome domain that is homologous to the cytochrome domain of cellobiose dehydrogenase (CDH; EC 1.1.99.18) from the wood-rotting basidiomycete Phanerochaete chrysosporium, a C-terminal family 1-type carbohydrate-binding module, and a novel central catalytic domain containing PQQ as a cofactor. Here, we describe the biochemical and electrochemical characterization of recombinant CcPDH. UV-vis and resonance Raman spectroscopic studies clearly reveal characteristics of a 6-coordinated low-spin heme b in both the ferric and ferrous states, as well as intramolecular electron transfer from the PQQ to heme b. Moreover, the formal potential of the heme was evaluated to be 130 mV vs. NHE by cyclic voltammetry. These results indicate that the cytochrome domain of CcPDH possesses similar biophysical properties to that in CDH. A comparison of the conformations of monosaccharides as substrates and the associated catalytic efficiency (kcat/Km) of CcPDH indicates that the enzyme prefers monosaccharides with equatorial C-2, C-3 hydroxyl groups and an axial C-4 hydroxyl group in the 1C4 chair conformation. Furthermore, a binding study shows a high binding affinity of CcPDH for cellulose, suggesting that CcPDH function is related to the enzymatic degradation of plant cell wall. PMID:25679509

  19. The Effects of Melatonin on Brain Arginine Vasotocin: Relationship with Sex and Seasonal Differences in Melatonin Receptor Type 1 in Green Treefrogs (Hyla cinerea).

    PubMed

    Howard, C M; Lutterschmidt, D I

    2015-08-01

    The neuroendocrine mechanisms by which animals synchronise their physiological state with environmental cues are vital to timing life-history events appropriately. One important endocrine transducer of environmental cues in vertebrates is the pineal hormone melatonin, the secretion of which is directly sensitive to photoperiod and temperature. Melatonin modulates arginine vasotocin (AVT)-immunoreactive (-IR) cell number in the brain of green treefrogs (Hyla cinerea) during the summer breeding season, and this modulation is sexually dimorphic. In the present study, we investigated whether the influence of melatonin on vasotocin varies seasonally. We show that treatment of nonreproductive male green treefrogs with melatonin-filled silastic implants for 4 weeks during the winter does not alter vasotocin-IR cell number in any brain region (i.e. nucleus accumbens, amygdala, preoptic area, suprachiasmatic nucleus or ventral hypothalamus). Taken together, these results suggest that the influence of melatonin on AVT is associated with sex and seasonal variation in melatonin receptor expression. We tested this hypothesis by using immunohistochemistry to characterise the distribution of melatonin receptor type 1 (MT1, also known as Mel1a) in the brain of reproductive and nonreproductive male and female frogs. We quantified MT1-IR cell number in regions known to contain AVT cell populations. Reproductive males had significantly more MT1-IR cells than nonreproductive males in all brain regions, including the combined nucleus accumbens, diagonal band of Broca and septum, striatum, amygdala, combined preoptic area and suprachiasmatic nucleus, as well as the ventral hypothalamus. In the accumbens region, where the effect of melatonin on AVT is known to be sexually dimorphic, males had significantly more MT1-IR cells than females during the summer breeding season. Based on these findings, we suggest that MT1 plays a role in mediating the interactions between melatonin and

  20. Characterization of a Novel PQQ-Dependent Quinohemoprotein Pyranose Dehydrogenase from Coprinopsis cinerea Classified into Auxiliary Activities Family 12 in Carbohydrate-Active Enzymes

    PubMed Central

    Takeda, Kouta; Matsumura, Hirotoshi; Ishida, Takuya; Samejima, Masahiro; Ohno, Hiroyuki; Yoshida, Makoto; Igarashi, Kiyohiko; Nakamura, Nobuhumi

    2015-01-01

    The basidiomycete Coprinopsis cinerea contains a quinohemoprotein (CcPDH named as CcSDH in our previous paper), which is a new type of pyrroloquinoline-quinone (PQQ)-dependent pyranose dehydrogenase and is the first found among all eukaryotes. This enzyme has a three-domain structure consisting of an N-terminal heme b containing a cytochrome domain that is homologous to the cytochrome domain of cellobiose dehydrogenase (CDH; EC 1.1.99.18) from the wood-rotting basidiomycete Phanerochaete chrysosporium, a C-terminal family 1-type carbohydrate-binding module, and a novel central catalytic domain containing PQQ as a cofactor. Here, we describe the biochemical and electrochemical characterization of recombinant CcPDH. UV-vis and resonance Raman spectroscopic studies clearly reveal characteristics of a 6-coordinated low-spin heme b in both the ferric and ferrous states, as well as intramolecular electron transfer from the PQQ to heme b. Moreover, the formal potential of the heme was evaluated to be 130 mV vs. NHE by cyclic voltammetry. These results indicate that the cytochrome domain of CcPDH possesses similar biophysical properties to that in CDH. A comparison of the conformations of monosaccharides as substrates and the associated catalytic efficiency (kcat/Km) of CcPDH indicates that the enzyme prefers monosaccharides with equatorial C-2, C-3 hydroxyl groups and an axial C-4 hydroxyl group in the 1C4 chair conformation. Furthermore, a binding study shows a high binding affinity of CcPDH for cellulose, suggesting that CcPDH function is related to the enzymatic degradation of plant cell wall. PMID:25679509

  1. Microwave-assisted Synthesis and antifungal activity of coumarin[8,7-e][1,3]oxazine derivatives.

    PubMed

    Zhang, Ming-Zhi; Zhang, Rong-Rong; Yin, Wen-Zheng; Yu, Xiang; Zhang, Ya-Ling; Liu, Pin; Gu, Yu-Cheng; Zhang, Wei-Hua

    2016-08-01

    The synthesis of novel coumarin[8,7-e][1,3]oxazine derivatives through a microwave-assisted three-component one-pot Mannich reaction is described in this study. All the target compounds were evaluated in vitro for their antifungal activity against Botrytis cinerea, Colletotrichum capsici, Alternaria solani, Gibberella zeae, Rhizoctonia solani, and Alternaria mali. The preliminary bioassays showed that 5e, 5m, and 5s exhibited good antifungal activity and the most active compound was 5m with an [Formula: see text] value as low as 2.1 nM against Botrytis cinerea. PMID:26880591

  2. First description of the adult stage of Clinostomum cutaneum Paperna, 1964 (Digenea: Clinostomidae) from grey herons Ardea cinerea L. and a redescription of the metacercaria from the Nile tilapia Oreochromis niloticus niloticus (L.) in Kenya.

    PubMed

    Gustinelli, Andrea; Caffara, Monica; Florio, Daniela; Otachi, Elick O; Wathuta, Euty M; Fioravanti, Maria L

    2010-05-01

    The combined use of morphological and molecular studies allowed for the first time the recognition and description of the adult stage of Clinostomum cutaneum Paperna, 1964 from the grey heron Ardea cinerea L. in Kenya. A redescription of the metacercaria that infect Nile tilapia Oreochromis niloticus niloticus (L.) from the same aquatic environment is also presented. C. cutaneum differs from all other species of Clinostomum Leidy, 1856 in the shape of its uterus. Sequencing the rRNA confirmed the morphological similarity between adults from the grey heron and the metacercarial stage from tilapia, and a level of genetic similarity with the other previously sequenced Clinostomum spp. was observed. The need for a reorganisation of Clinostomum using both morphological and molecular methods is highlighted. PMID:20401577

  3. Identification of differentially expressed genes associated with changes in the morphology and pathogenicity of Pichia fermentans on apple and peach fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pichia fermentans (strain DISAABA 726) is an effective biocontrol agent against Monilinia fructicola or Botrytis cinerea when inoculated in artificially wounded apple fruit but is an aggressive pathogen when inoculated on wounded peach fruit, causing severe fruit decay. P. fermentans grows as a bud...

  4. Antagonistic activity and mechanisms of Bacillus subtilis SB1 against Ralstonia solanacearum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A potential biocontrol agent of bacterial wilt, Bacillus subtilis SB1, showed a broad-spectrum of antimicrobial activity in vitro experiments. In addition to Ralstonia solanacearum, strain SB1 inhibited the growth of many other plant pathogens, including Fusarium oxysporum, Botrytis cinerea, Phytoph...

  5. Recent advances on the use of natural and safe alternatives to conventional methods to control postharvest gray mold of table grapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gray mold, caused by Botrytis cinerea, is the main postharvest decay of table grapes. It can develop in the vineyard and spread rapidly among berries after harvest, during long distant transport, cold storage and shelf life. In conventional agriculture, bunches are sprayed with fungicides after flow...

  6. Effects of pre- and post-harvest application of selenium on inducing disease resistance and selenium accumulation in fruits

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis cinerea, a ubiquitous fungal pathogen, causes severe damage (gray mold rot) on a large number of economically important fruits, vegetables, and ornamental crops at both pre- and post-harvest, which renders fruits unmarketable. Penicillium expansum is a widely spread fungal pathogen that cau...

  7. Application of low concentrations of ozone during the cold storage of table grapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The control of postharvest decay of table grapes, caused by Botrytis cinerea and other pathogens, by ozone was evaluated in chambers and commercial storage facilities. Ozone at 0.100 µL/L or higher inhibited the spread of gray mold among stored grapes. Ozone diffusion into many types of commercial p...

  8. Disinfection of immature fruit extends shelf-life in Florida strawberries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fresh market Florida strawberries (Fragaria x ananassa) are field packed and shipped without further processing. Keeping the fruit cool (~0 C (32 F)) helps maintain the quality of the strawberries but controlled temperatures are not always accessible. Organisms such as Botrytis cinerea (grey mold)...

  9. Influence of fumigation with high concentrations of ozone gas on postharvest gray mold, quality, and fungicide residues on table grapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To control postharvest decay table grapes are commercially fumigated with sulfur dioxide. We evaluated ozone (O3) fumigation with up to10,000 microl/l of ozone for up to 2 h to control postharvest gray mold of table grapes caused by Botrytis cinerea. Fumigation for 1 h with 2,500 or 5,000 microl/l o...

  10. The relationship of biofilm production to biocontrol activity of Burkholderia pyrrocinia FP62

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Foliar biocontrol agent (BCA) efficacy is often inconsistent due to poor colonization and survival on plant surfaces. Burkholderia pyrrocinia FP62, a superior leaf colonist and BCA of Botrytis cinerea, forms unsaturated biofilms on plant surfaces. To determine the relationship between biocontrol act...

  11. Use of jasmonic acid and salicylic acid to inhibit growth of sugarbeet storage rot pathogens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Jasmonic acid (JA) and salicylic acid (SA) are endogenous plant hormones that induce native plant defense responses and provide protection against a wide range of diseases. Previously, JA, applied after harvest, was shown to protect sugarbeet roots against the storage pathogens, Botrytis cinerea, P...

  12. Enhancing Florida strawberry shelf life using peroxiacetic acid pre-harvest treatments and postharvest edible coatings

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Strawberry is an important fruit crop in Florida. Yearly losses can be attributed to pre- and postharvest decay caused by Botrytis cinerea, and postharvest decay due primarily to Rhizopus stolonifer. Postharvest applications of edible coatings and sanitizers were tested. Among them, a medium-dens...

  13. Biotransformations of 6',7'-dihydroxybergamottin and 6',7'-epoxybergamottin by the citrus-pathogenic fungi diminish cytochrome P450 3A4 inhibitory activity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium digitatum, as well as five other citrus pathogenic species, (P. ulaiense Link, Geotrichum citri Link, Botrytis cinerea P. Micheli ex Pers., Lasiodiplodia theobromae (Pat.)Griffon & Maubl. and Phomopsis citri (teleomorph Diaporthe citri)) was observed to convert 6',7'-epoxybergamottin (1)...

  14. Evaluation under commercial conditions of the application of continuous, low concentrations of ozone during the cold storage of table grapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botrytis cinerea causes gray mold, a postharvest disease of table grapes. The ability of ozone (O3) in air to inhibit gray mold in stored grapes was reported in chamber studies, but O3 needed evaluation under commercial conditions. Ozone merits attention because it is pesticide-residue free and all...

  15. Sulfur dioxide-releasing perforated plastic liners to control postharvest gray mold of Redglobe table grapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    "Gray mold, caused by Botrytis cinerea, limits the duration of table grape storage. Periodic sulfur dioxide (SO2) fumigation and in-package SO2 generating pads are two common strategies that protect grapes after harvest. Our objectives were to compare the effectiveness of packaging Redglobe grapes i...

  16. Complete genome sequence of Bacillus subtilis BSD-2, a microbial germicide isolated from cultivated cotton.

    PubMed

    Liu, Hongwei; Yin, Shuli; An, Likang; Zhang, Genwei; Cheng, Huicai; Xi, Yanhua; Cui, Guanhui; Zhang, Feiyan; Zhang, Liping

    2016-07-20

    Bacillus subtilis BSD-2, isolated from cotton (Gossypium spp.), had strong antagonistic activity to Verticillium dahlia Kleb and Botrytis cinerea. We sequenced and annotated the BSD-2 complete genome to help us the better use of this strain, which has surfactin, bacilysin, bacillibactin, subtilosin A, Tas A and a potential class IV lanthipeptide biosynthetic pathways. PMID:27184432

  17. Integration of continuous biofumigation with Muscodor albus with pre-cooling fumigation with ozone or sulfur dioxide to control postharvest gray mold of table grapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sulfur dioxide (SO2) fumigation controls postharvest decay of commercially stored table grapes. To develop an alternative to SO2, fumigation with up to 10,000 micro-l/l ozone (O3) for up to 2 h was applied to control postharvest gray mold caused by Botrytis cinerea. O3 was effective when grapes were...

  18. Evaluation of sulfur dioxide-generating pads and modified atmosphere packaging for control of postharvest diseases in blueberries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Postharvest diseases are a limiting factor of storage and shelf life of blueberries. Gray mold caused by Botrytis cinerea is one of the most important postharvest diseases in blueberries grown in California. In this study, we evaluated the effects of sulfur dioxide (SO2)-generating pads (designated ...

  19. Identification of genes involved in the pathogenic interaction between an antagonistic strain of Pichia fermentans and peach fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A biofilm-forming strain of Pichia fermentans was very effective in reducing brown rot and grey rot in artificially wounded apple fruit when co-inoculated with either Monilinia fructicola or Botrytis cinerea, respectively. The same strain, however, was an aggressive pathogen when inoculated on peach...

  20. Differential gene expression during the pathogenic interaction between Pichia fermentans and peach fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A biofilm-forming strain of Pichia fermentans was found to be a very strong antagonist against brown rot and grey mold in artificially wounded apple fruit when co-inoculated with either Monilinia fructicola or Botrytis cinerea, respectively. The same strain of yeast; however, was an aggressive path...

  1. Antifungal effect of some spice hydrosols.

    PubMed

    Boyraz, Nuh; Ozcan, Musa

    2005-12-01

    The antifungal effects of rosemary, cumin, sater (savory), basil and pickling herb hydrosols were investigated against Rhizoctonia solani, Fusarium oxysporum f. sp tulipae, Botrytis cinerea and Alternaria citri. Hydrosols of sater and pickling herb showed the most relevant fungicidal activity. PMID:16243447

  2. Preharvest peroxyacetic acid sprays slow decay and extend shelf-life of strawberries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Strawberry is the most important berry crop in Florida. Yearly losses can be attributed to pre and postharvest decay caused by Botrytis cinerea P. Micheli ex Pers., and postharvest decay due primarily to Rhizopus stolonifer (Ehrenb. ex Fr.) Vuillemin. In this study the sanitizer peroxyacetic acid ...

  3. A framework map from grapevine V3125 (Vitis vinifera 'Schiava grossa' x 'Riesling') x rootstock cultivar 'Börner' (Vitis riparia x Vitis cinerea) to localize genetic determinants of phylloxera root resistance.

    PubMed

    Zhang, Junke; Hausmann, Ludger; Eibach, Rudolf; Welter, Leocir J; Töpfer, Reinhard; Zyprian, Eva M

    2009-10-01

    Grapevine rootstock cultivar 'Börner' is a hybrid of Vitis riparia and Vitis cinerea Arnold that shows high resistance to phylloxera (Daktulosphaira vitifoliae Fitch). To localize the determinants of phylloxera root resistance, the susceptible grapevine V3125 (Vitis vinifera 'Schiava grossa' x 'Riesling') was crossed to 'Börner'. Genetic framework maps were built from the progeny. 235 microsatellite markers were placed on the integrated parental map. They cover 1,155.98 cM on 19 linkage groups with an average marker distance of 4.8 cM. Phylloxera resistance was scored by counting nodosities after inoculation of the root system. Progeny plants were triplicated and experimentally infected in 2 years. A scan of the genetic maps indicated a quantitative trait locus on linkage group 13. This region was targeted by six microsatellite-type markers newly developed from the V. vinifera model genome sequence. Two of these appear closely linked to the trait, and can be useful for marker-assisted breeding. PMID:19626311

  4. Biological control of the grapevine diseases 'grey mold' and 'powdery mildew' by Bacillus B27 and B29 strains.

    PubMed

    Maachia, Ben; Rafik, Errakhi; Chérif, M; Nandal, Preeti; Mohapatra, Trupti; Bernard, Paul

    2015-02-01

    Uncinula necator and Botrytis cinerea are the most destructive pathogens of the grapevine in Tunisia and elsewhere. We used two strains of Bacillus subtilis group, B27 and B29 to control powdery mildew and the grey mold disease of the grapevine. Green house experiments showed that B29 and B27 strains of the bacteria efficiently reduced the severity of powdery mildew up to 50% and 60%, respectively. Further, they decreased Botrytis cinerea development on grape leaf by 77% and 99%, respectively. The mode of action has been shown to be chitinolytic. These two bacteria showed significant production of total proteins discharged into the culture medium. Determination of some chitinolytic enzymes revealed the involvement of N-acetyl glucosaminidase (Nagase), the chitin-1,4-chitobiosidase (Biase) and endochitinase in degrading the mycelium of B. cinerea. PMID:25757242

  5. A mutation in the Cc.arp9 gene encoding a putative actin-related protein causes defects in fruiting initiation and asexual development in the agaricomycete Coprinopsis cinerea.

    PubMed

    Nakazawa, Takehito; Ando, Yuki; Hata, Takeshi; Nakahori, Kiyoshi

    2016-08-01

    Agaricomycetes exhibit a remarkable morphological differentiation from vegetative mycelia to huge fruiting bodies. To investigate the molecular mechanism underlying the fruiting body development, we have isolated and characterized many Coprinopsis cinerea mutant strains defective in fruiting initiation to date. Dikaryon formation in agaricomycetes, which is followed by fruiting development, is governed by the mating type loci, A and B. Recently, mutations in the Cc.snf5 gene, which encodes a putative component of the chromatin remodeling complex switch/sucrose non-fermentable (SWI/SNF), were shown to cause defects in A-regulated clamp cell morphogenesis, as well as in fruiting initiation. Here, we demonstrate that Cc.arp9, which encodes a putative actin-related protein associated with two chromatin remodeling complexes, SWI/SNF and remodels the structure of chromatin (RSC), is also essential for fruiting initiation. In contrast to Cc.snf5 mutants, Cc.arp9 mutants were not defective in clamp cell formation. The effects of mutations in Cc.arp9 and Cc.snf5 on oidia production and the transcriptional expression levels of clp1 and pcc1, which are under the control of the A gene, were also examined. These indicated that Cc.Snf5 is involved in A-regulated pathways, whereas Cc.Arp9 is not apparently. Cc.arp9/Cc.snf5 double-gene disruptants were generated and their phenotypes were analyzed, which suggested a complicated developmental regulation mechanism mediated by chromatin remodeling. PMID:26746642

  6. Can the grey mould disease of the grape-vine be controlled by yeast?

    PubMed

    Masih, E I; Alie, I; Paul, B

    2000-08-15

    Botrytis cinerea has been found to be highly pathogenic to 'Chardonnay' and 'Pinot noir' cultivars of the grape-vine producing the characteristic grey mould symptoms within 7 days of inoculation to the vitro-plants. The yeast Pichia anomala (strain FY-102), isolated from apple skin, was found to be antagonistic to B. cinerea as it completely inhibited the appearance of the grey mould symptoms when grown together. The yeast was responsible for morphological changes such as coagulation and leakage of the cytoplasm of B. cinerea. The pathogen, when applied together with P. anomala, failed to bring about the grey mould symptoms on the grape-vine, suggesting that the yeast could control the expression of this disease. An account of the interaction between B. cinerea and P. anomala, as well as the sequences of the complete ITS region of the ribosomal DNA of the yeast are described here. PMID:10930744

  7. Fungal small RNAs suppress plant immunity by hijacking host RNA interference pathways.

    PubMed

    Weiberg, Arne; Wang, Ming; Lin, Feng-Mao; Zhao, Hongwei; Zhang, Zhihong; Kaloshian, Isgouhi; Huang, Hsien-Da; Jin, Hailing

    2013-10-01

    Botrytis cinerea, the causative agent of gray mold disease, is an aggressive fungal pathogen that infects more than 200 plant species. Here, we show that some B. cinerea small RNAs (Bc-sRNAs) can silence Arabidopsis and tomato genes involved in immunity. These Bc-sRNAs hijack the host RNA interference (RNAi) machinery by binding to Arabidopsis Argonaute 1 (AGO1) and selectively silencing host immunity genes. The Arabidopsis ago1 mutant exhibits reduced susceptibility to B. cinerea, and the B. cinerea dcl1 dcl2 double mutant that can no longer produce these Bc-sRNAs displays reduced pathogenicity on Arabidopsis and tomato. Thus, this fungal pathogen transfers "virulent" sRNA effectors into host plant cells to suppress host immunity and achieve infection, which demonstrates a naturally occurring cross-kingdom RNAi as an advanced virulence mechanism. PMID:24092744

  8. Perception of soft mechanical stress in Arabidopsis leaves activates disease resistance

    PubMed Central

    2013-01-01

    Background In a previous study we have shown that wounding of Arabidopsis thaliana leaves induces a strong and transient immunity to Botrytis cinerea, the causal agent of grey mould. Reactive oxygen species (ROS) are formed within minutes after wounding and are required for wound–induced resistance to B. cinerea. Results In this study, we have further explored ROS and resistance to B. cinerea in leaves of A. thaliana exposed to a soft form of mechanical stimulation without overt tissue damage. After gentle mechanical sweeping of leaf surfaces, a strong resistance to B. cinerea was observed. This was preceded by a rapid change in calcium concentration and a release of ROS, accompanied by changes in cuticle permeability, induction of the expression of genes typically associated with mechanical stress and release of biologically active diffusates from the surface. This reaction to soft mechanical stress (SMS) was fully independent of jasmonate (JA signaling). In addition, leaves exposed soft mechanical stress released a biologically active product capable of inducing resistance to B. cinerea in wild type control leaves. Conclusion Arabidopsis can detect and convert gentle forms of mechanical stimulation into a strong activation of defense against the virulent fungus B. cinerea. PMID:24033927

  9. Tetraphenylphosphonium allyldithiocarbimates derived from Morita-Baylis-Hillman adducts: Synthesis, characterization, crystal structure and antifungal activity

    NASA Astrophysics Data System (ADS)

    Tavares, Eder C.; Rubinger, Mayura M. M.; Filho, Eclair V.; Oliveira, Marcelo R. L.; Piló-Veloso, Dorila; Ellena, Javier; Guilardi, Silvana; Souza, Rafael A. C.; Zambolim, Laércio

    2016-02-01

    Botrytis blight is a very destructive disease caused by Botrytis spp., infecting flowers, trees, vegetables and fruits. Twelve new compounds were prepared by the reaction of potassium N-aryl-sulfonyldithiocarbimates with Morita-Baylis-Hillman derivatives bearing phenyl and furyl groups. These are the first examples of allyldithiocarbimate anions and were isolated as tetraphenylphosphonium salts. The new compounds were characterized by HRMS, NMR and Infrared spectroscopy. Further, the structures of three allyldithiocarbimates were determined by single crystal X-ray diffraction. The compounds are isostructural and crystallize in the space group P21/c of the monoclinic system, and the allyldithiocarbimate anions present Z configuration. All the compounds were active against Botrytis cinerea. The best results were achieved with the tetraphenylphosphonium (Z)-3-(furan-2-yl)-2-(methoxycarbonyl)allyl-(4-chlorophenylsulfonyl)dithiocarbimate (IC50 38 μM).

  10. Antifungal and Cytotoxic Assessment of Lapachol Derivatives Produced by Fungal Biotransformation.

    PubMed

    Silva, Eliane O; Ruano-González, Antonio; dos Santos, Raquel A; Sánchez-Maestre, Rosario; Furtado, Niege A J C; Collado, Isidro G; Aleu, Josefina

    2016-01-01

    In the screening for biological active compounds, the biotransformation processes catalyzed by filamentous fungi are useful because they can provide information about the possible appearance of toxic metabolites after oral administration and also generate new leads. In this paper, biotransformation of lapachol (1) by three fungal strains, Mucor circinelloides NRRL3631, Botrytis cinerea UCA992 and Botrytis cinerea 2100, has been investigated for the first time. Lapachol (1) was biotransformed into avicequinone-A (2) by M circinelloides, 3'-hydroxylapachol (3) by B. cinerea, and into dehydro-α-lapachone (4) by both fungi. All these compounds were evaluated for their cytotoxic activities. The metabolite 2 displayed non-selective cytotoxicity against tumor and normal cell lines, 3 did not show cytotoxicity against the same cells, while 4 showed higher cytotoxicity against cancer cell lines than lapachol (1). The transformation of 1 into harmless and reactive metabolites evidences the importance of the evaluation of drug metabolism in the drug discovery process. Antifungal potential of lapachol (1) and its metabolites 2 and 4 against B. cinerea has also been evaluated. Dehydro-α-lapachone (4) has been shown to be less toxic to fungal growth than lapachol (1), which indicates a detoxification mechanism of the phytopathogen. PMID:26996030

  11. Making healthier or killing enemies? Bacterial volatile-elicited plant immunity plays major role upon protection of Arabidopsis than the direct pathogen inhibition.

    PubMed

    Sharifi, Rouhallah; Ryu, Choong-Min

    2016-01-01

    Bacterial volatiles protect plants either by directly inhibiting a pathogenic fungus or by improving the defense capabilities of plants. The effect of bacterial volatiles on fungal growth was dose-dependent. A low dosage did not have a noticeable effect on Botrytis cinerea growth and development, but was sufficient to elicit induced resistance in Arabidopsis thaliana. Bacterial volatiles displayed negative effects on biofilm formation on a polystyrene surface and in in planta leaf colonization of B. cinerea. However, bacterial volatile-mediated induced resistance was the major mechanism mediating protection of plants from B. cinerea. It was responsible for more than 90% of plant protection in comparison with direct fungal inhibition. Our results broaden our knowledge of the role of bacterial volatiles in plant protection. PMID:27574539

  12. Synthesis and antifungal activity evaluation of new heterocycle containing amide derivatives.

    PubMed

    Wang, Xuesong; Gao, Sumei; Yang, Jian; Gao, Yang; Wang, Ling; Tang, Xiaorong

    2016-01-01

    A series of heterocycle containing amide derivatives (1-28) were synthesised by the combination of acyl chlorides (1a, 2a) and heterocyclic/homocyclic ring containing amines, and their in vitro antifungal activity was evaluated against five plant pathogenic fungi, namely Gibberella zeae, Helminthosporium maydis, Rhizoctonia solani, Botrytis cinerea and Sclerotinia sclerotiorum. Results of antifungal activity analysis indicated that some of the products showed good to excellent antifungal activity, as compound 2 showed excellent activity against G. zeae and R. solani and potent activity against H. maydi, B. cinerea and S. sclerotiorum, and compounds 1, 8 and 10 also displayed excellent antifungal potential against H. maydi, B. cinerea and S. sclerotiorum and good activity against R. solani when compared with the standard carbendazim. PMID:26140452

  13. Making healthier or killing enemies? Bacterial volatile-elicited plant immunity plays major role upon protection of Arabidopsis than the direct pathogen inhibition

    PubMed Central

    Sharifi, Rouhallah; Ryu, Choong-Min

    2016-01-01

    ABSTRACT Bacterial volatiles protect plants either by directly inhibiting a pathogenic fungus or by improving the defense capabilities of plants. The effect of bacterial volatiles on fungal growth was dose-dependent. A low dosage did not have a noticeable effect on Botrytis cinerea growth and development, but was sufficient to elicit induced resistance in Arabidopsis thaliana. Bacterial volatiles displayed negative effects on biofilm formation on a polystyrene surface and in in planta leaf colonization of B. cinerea. However, bacterial volatile-mediated induced resistance was the major mechanism mediating protection of plants from B. cinerea. It was responsible for more than 90% of plant protection in comparison with direct fungal inhibition. Our results broaden our knowledge of the role of bacterial volatiles in plant protection. PMID:27574539

  14. Analysis of Clonostachys rosea-Induced Resistance to Tomato Gray Mold Disease in Tomato Leaves

    PubMed Central

    Guan, Xin; Chen, Xiuling; Chen, Hongyu; Zhang, Jian; Zhang, Junfeng; Li, Jingfu; Yang, Yijun; Wang, Aoxue

    2014-01-01

    Tomato gray mold disease, caused by Botrytis cinerea, is a serious disease in tomato. Clonostachys rosea is an antagonistic microorganism to B. cinerea. To investigate the induced resistance mechanism of C. rosea, we examined the effects of these microorganisms on tomato leaves, along with changes in the activities of three defense enzymes (PAL, PPO, GST), second messengers (NO, H2O2, O2−) and phytohormones (IAA, ABA, GA3, ZT, MeJA, SA and C2H4). Compared to the control, all treatments induced higher levels of PAL, PPO and GST activity in tomato leaves and increased NO, SA and GA3 levels. The expression of WRKY and MAPK, two important transcription factors in plant disease resistance, was upregulated in C. rosea- and C. rosea plus B. cinerea-treated samples. Two-dimensional gel electrophoresis analysis showed that two abundant proteins were present in the C. rosea plus B. cinerea-treated samples but not in the other samples. These proteins were determined (by mass spectrum analysis) to be LEXYL2 (β-xylosidase) and ATP synthase CF1 alpha subunit. Therefore, C. rosea plus B. cinerea treatment induces gray mold resistance in tomato. This study provides a basis for elucidating the mechanism of C. rosea as a biocontrol agent. PMID:25061981

  15. The Effects of Glucosinolates and Their Breakdown Products on Necrotrophic Fungi

    PubMed Central

    Barda, Omer; Levy, Maggie

    2013-01-01

    Glucosinolates are a diverse class of S- and N-containing secondary metabolites that play a variety of roles in plant defense. In this study, we used Arabidopsis thaliana mutants that contain different amounts of glucosinolates and glucosinolate-breakdown products to study the effects of these phytochemicals on phytopathogenic fungi. We compared the fungus Botrytis cinerea, which infects a variety of hosts, with the Brassicaceae-specific fungus Alternaria brassicicola. B. cinerea isolates showed variable composition-dependent sensitivity to glucosinolates and their hydrolysis products, while A. brassicicola was more strongly affected by aliphatic glucosinolates and isothiocyanates as decomposition products. We also found that B. cinerea stimulates the accumulation of glucosinolates to a greater extent than A. brassicicola. In our work with A. brassicicola, we found that the type of glucosinolate-breakdown product is more important than the type of glucosinolate from which that product was derived, as demonstrated by the sensitivity of the Ler background and the sensitivity gained in Col-0 plants expressing epithiospecifier protein both of which accumulate simple nitrile and epithionitriles, but not isothiocyanates. Furthermore, in vivo, hydrolysis products of indole glucosinolates were found to be involved in defense against B. cinerea, but not in the host response to A. brassicicola. We suggest that the Brassicaceae-specialist A. brassicicola has adapted to the presence of indolic glucosinolates and can cope with their hydrolysis products. In contrast, some isolates of the generalist B. cinerea are more sensitive to these phytochemicals. PMID:23940639

  16. Molecular and Functional Characterization of a Polygalacturonase-Inhibiting Protein from Cynanchum komarovii That Confers Fungal Resistance in Arabidopsis

    PubMed Central

    Liu, Nana; Ma, Xiaowen; Zhou, Sihong; Wang, Ping; Sun, Yun; Li, Xiancai; Hou, Yuxia

    2016-01-01

    Compliance with ethical standards: This study did not involve human participants and animals, and the plant of interest is not an endangered species. Polygalacturonase-inhibiting proteins (PGIPs) are leucine-rich repeat proteins that plants produce against polygalacturonase, a key virulence agent in pathogens. In this paper, we cloned and purified CkPGIP1, a gene product from Cynanchum komarovii that effectively inhibits polygalacturonases from Botrytis cinerea and Rhizoctonia solani. We found the expression of CkPGIP1 to be induced in response to salicylic acid, wounding, and infection with B. cinerea and R. solani. In addition, transgenic overexpression in Arabidopsis enhanced resistance against B. cinerea. Furthermore, CkPGIP1 obtained from transgenic Arabidopsis inhibited the activity of B. cinerea and R. solani polygalacturonases by 62.7–66.4% and 56.5–60.2%, respectively. Docking studies indicated that the protein interacts strongly with the B1-sheet at the N-terminus of the B. cinerea polygalacturonase, and with the C-terminus of the polygalacturonase from R. solani. This study highlights the significance of CkPGIP1 in plant disease resistance, and its possible application to manage fungal pathogens. PMID:26752638

  17. Molecular and Functional Characterization of a Polygalacturonase-Inhibiting Protein from Cynanchum komarovii That Confers Fungal Resistance in Arabidopsis.

    PubMed

    Liu, Nana; Ma, Xiaowen; Zhou, Sihong; Wang, Ping; Sun, Yun; Li, Xiancai; Hou, Yuxia

    2016-01-01

    Compliance with ethical standards: This study did not involve human participants and animals, and the plant of interest is not an endangered species. Polygalacturonase-inhibiting proteins (PGIPs) are leucine-rich repeat proteins that plants produce against polygalacturonase, a key virulence agent in pathogens. In this paper, we cloned and purified CkPGIP1, a gene product from Cynanchum komarovii that effectively inhibits polygalacturonases from Botrytis cinerea and Rhizoctonia solani. We found the expression of CkPGIP1 to be induced in response to salicylic acid, wounding, and infection with B. cinerea and R. solani. In addition, transgenic overexpression in Arabidopsis enhanced resistance against B. cinerea. Furthermore, CkPGIP1 obtained from transgenic Arabidopsis inhibited the activity of B. cinerea and R. solani polygalacturonases by 62.7-66.4% and 56.5-60.2%, respectively. Docking studies indicated that the protein interacts strongly with the B1-sheet at the N-terminus of the B. cinerea polygalacturonase, and with the C-terminus of the polygalacturonase from R. solani. This study highlights the significance of CkPGIP1 in plant disease resistance, and its possible application to manage fungal pathogens. PMID:26752638

  18. A Permeable Cuticle Is Associated with the Release of Reactive Oxygen Species and Induction of Innate Immunity

    PubMed Central

    L'Haridon, Floriane; Besson-Bard, Angélique; Binda, Matteo; Serrano, Mario; Abou-Mansour, Eliane; Balet, Francine; Schoonbeek, Henk-Jan; Hess, Stephane; Mir, Ricardo; Léon, José; Lamotte, Olivier; Métraux, Jean-Pierre

    2011-01-01

    Wounded leaves of Arabidopsis thaliana show transient immunity to Botrytis cinerea, the causal agent of grey mould. Using a fluorescent probe, histological staining and a luminol assay, we now show that reactive oxygen species (ROS), including H2O2 and O2−, are produced within minutes after wounding. ROS are formed in the absence of the enzymes Atrboh D and F and can be prevented by diphenylene iodonium (DPI) or catalase. H2O2 was shown to protect plants upon exogenous application. ROS accumulation and resistance to B. cinerea were abolished when wounded leaves were incubated under dry conditions, an effect that was found to depend on abscisic acid (ABA). Accordingly, ABA biosynthesis mutants (aba2 and aba3) were still fully resistant under dry conditions even without wounding. Under dry conditions, wounded plants contained higher ABA levels and displayed enhanced expression of ABA-dependent and ABA-reporter genes. Mutants impaired in cutin synthesis such as bdg and lacs2.3 are already known to display a high level of resistance to B. cinerea and were found to produce ROS even when leaves were not wounded. An increased permeability of the cuticle and enhanced ROS production were detected in aba2 and aba3 mutants as described for bdg and lacs2.3. Moreover, leaf surfaces treated with cutinase produced ROS and became more protected to B. cinerea. Thus, increased permeability of the cuticle is strongly linked with ROS formation and resistance to B. cinerea. The amount of oxalic acid, an inhibitor of ROS secreted by B. cinerea could be reduced using plants over expressing a fungal oxalate decarboxylase of Trametes versicolor. Infection of such plants resulted in a faster ROS accumulation and resistance to B. cinerea than that observed in untransformed controls, demonstrating the importance of fungal suppression of ROS formation by oxalic acid. Thus, changes in the diffusive properties of the cuticle are linked with the induction ROS and attending innate defenses. PMID

  19. Novel components of leaf bacterial communities of field-grown tomato plants and their potential for plant growth promotion and biocontrol of tomato diseases.

    PubMed

    Romero, Fernando M; Marina, María; Pieckenstain, Fernando L

    2016-04-01

    This work aimed to characterize potentially endophytic culturable bacteria from leaves of cultivated tomato and analyze their potential for growth promotion and biocontrol of diseases caused by Botrytis cinerea and Pseudomonas syringae. Bacteria were obtained from inner tissues of surface-disinfected tomato leaves of field-grown plants. Analysis of 16S rRNA gene sequences identified bacterial isolates related to Exiguobacterium aurantiacum (isolates BT3 and MT8), Exiguobacterium spp. (isolate GT4), Staphylococcus xylosus (isolate BT5), Pantoea eucalypti (isolate NT6), Bacillus methylotrophicus (isolate MT3), Pseudomonas veronii (isolates BT4 and NT2), Pseudomonas rhodesiae (isolate BT2) and Pseudomonas cichorii (isolate NT3). After seed inoculation, BT2, BT4, MT3, MT8, NT2 and NT6 were re-isolated from leaf extracts. NT2, BT2, MT3 and NT6 inhibited growth of Botrytis cinerea and Pseudomonas syringae pv. tomato in vitro, produced antimicrobial compounds and reduced leaf damage caused by B. cinerea. Some of these isolates also promoted growth of tomato plants, produced siderophores, the auxin indole-3-acetic and solubilized inorganic phosphate. Thus, bacterial communities of leaves from field-grown tomato plants were found to harbor potentially endophytic culturable beneficial bacteria capable of antagonizing pathogenic microorganisms and promoting plant growth, which could be used as biological control agents and biofertilizers/biostimulators for promotion of tomato plant growth. PMID:26654914

  20. Tomato Protein Kinase 1b Mediates Signaling of Plant Responses to Necrotrophic Fungi and Insect Herbivory[W

    PubMed Central

    AbuQamar, Synan; Chai, Mao-Feng; Luo, Hongli; Song, Fengming; Mengiste, Tesfaye

    2008-01-01

    The tomato protein kinase 1 (TPK1b) gene encodes a receptor-like cytoplasmic kinase localized to the plasma membrane. Pathogen infection, mechanical wounding, and oxidative stress induce expression of TPK1b, and reducing TPK1b gene expression through RNA interference (RNAi) increases tomato susceptibility to the necrotrophic fungus Botrytis cinerea and to feeding by larvae of tobacco hornworm (Manduca sexta) but not to the bacterial pathogen Pseudomonas syringae. TPK1b RNAi seedlings are also impaired in ethylene (ET) responses. Notably, susceptibility to Botrytis and insect feeding is correlated with reduced expression of the proteinase inhibitor II gene in response to Botrytis and 1-aminocyclopropane-1-carboxylic acid, the natural precursor of ET, but wild-type expression in response to mechanical wounding and methyl-jasmonate. TPK1b functions independent of JA biosynthesis and response genes required for resistance to Botrytis. TPK1b is a functional kinase with autophosphorylation and Myelin Basis Protein phosphorylation activities. Three residues in the activation segment play a critical role in the kinase activity and in vivo signaling function of TPK1b. In sum, our findings establish a signaling role for TPK1b in an ET-mediated shared defense mechanism for resistance to necrotrophic fungi and herbivorous insects. PMID:18599583

  1. Microwave-assisted synthesis and antifungal activity of novel coumarin derivatives: Pyrano[3,2-c]chromene-2,5-diones.

    PubMed

    Zhang, Rong-Rong; Liu, Jia; Zhang, Yu; Hou, Meng-Qing; Zhang, Ming-Zhi; Zhou, Fenger; Zhang, Wei-Hua

    2016-06-30

    A series of novel fused coumarin analogues pyrano[3,2-c]chromene-2,5-diones have been synthesized through an optimized microwave-assisted protocol. All target compounds were tested and evaluated for their antifungal activity against Botrytis cinerea, Colletotrichum copsica, Alternaria solani, Gibberella zeae and Rhizoctorzia solani. The bioassay results indicated that some of the compounds exhibited potent antifungal activities at concentration less than 50 ppm. For the compounds 5d, 6c and 7b, EC50 values against B. cinerea were as low as 0.141, 0.082 and 0.091 μM, respectively, which represents better antifungal activity than that of the commonly used fungicide Azoxystrobin. Compounds 5d (57%) and 6c (55%) also exhibited more effective control than Azoxystrobin (44%) against Colletotrichum capsica. PMID:27060759

  2. Different Reactive Oxygen Species Scavenging Properties of Flavonoids Determine Their Abilities to Extend the Shelf Life of Tomato1[OPEN

    PubMed Central

    Zhang, Yang; De Stefano, Rosalba; Robine, Marie; Butelli, Eugenio; Bulling, Katharina; Hill, Lionel; Rejzek, Martin; Martin, Cathie; Schoonbeek, Henk-jan

    2015-01-01

    The shelf life of tomato (Solanum lycopersicum) fruit is determined by the processes of overripening and susceptibility to pathogens. Postharvest shelf life is one of the most important traits for commercially grown tomatoes. We compared the shelf life of tomato fruit that accumulate different flavonoids and found that delayed overripening is associated with increased total antioxidant capacity caused by the accumulation of flavonoids in the fruit. However, reduced susceptibility to Botrytis cinerea, a major postharvest fungal pathogen of tomato, is conferred by specific flavonoids only. We demonstrate an association between flavonoid structure, selective scavenging ability for different free radicals, and reduced susceptibility to B. cinerea. Our study provides mechanistic insight into how flavonoids influence the shelf life, information that could be used to improve the shelf life of tomato and, potentially, other soft fruit. PMID:26082399

  3. Identification of Arabidopsis Candidate Genes in Response to Biotic and Abiotic Stresses Using Comparative Microarrays

    PubMed Central

    Sham, Arjun; Moustafa, Khaled; Al-Ameri, Salma; Al-Azzawi, Ahmed; Iratni, Rabah; AbuQamar, Synan

    2015-01-01

    Plants have evolved with intricate mechanisms to cope with multiple environmental stresses. To adapt with biotic and abiotic stresses, plant responses involve changes at the cellular and molecular levels. The current study was designed to investigate the effects of combinations of different environmental stresses on the transcriptome level of Arabidopsis genome using public microarray databases. We investigated the role of cyclopentenones in mediating plant responses to environmental stress through TGA (TGACG motif-binding factor) transcription factor, independently from jasmonic acid. Candidate genes were identified by comparing plants inoculated with Botrytis cinerea or treated with heat, salt or osmotic stress with non-inoculated or non-treated tissues. About 2.5% heat-, 19% salinity- and 41% osmotic stress-induced genes were commonly upregulated by B. cinerea-treatment; and 7.6%, 19% and 48% of genes were commonly downregulated by B. cinerea-treatment, respectively. Our results indicate that plant responses to biotic and abiotic stresses are mediated by several common regulatory genes. Comparisons between transcriptome data from Arabidopsis stressed-plants support our hypothesis that some molecular and biological processes involved in biotic and abiotic stress response are conserved. Thirteen of the common regulated genes to abiotic and biotic stresses were studied in detail to determine their role in plant resistance to B. cinerea. Moreover, a T-DNA insertion mutant of the Responsive to Dehydration gene (rd20), encoding for a member of the caleosin (lipid surface protein) family, showed an enhanced sensitivity to B. cinerea infection and drought. Overall, the overlapping of plant responses to abiotic and biotic stresses, coupled with the sensitivity of the rd20 mutant, may provide new interesting programs for increased plant resistance to multiple environmental stresses, and ultimately increases its chances to survive. Future research directions towards a

  4. Identification of Arabidopsis candidate genes in response to biotic and abiotic stresses using comparative microarrays.

    PubMed

    Sham, Arjun; Moustafa, Khaled; Al-Ameri, Salma; Al-Azzawi, Ahmed; Iratni, Rabah; AbuQamar, Synan

    2015-01-01

    Plants have evolved with intricate mechanisms to cope with multiple environmental stresses. To adapt with biotic and abiotic stresses, plant responses involve changes at the cellular and molecular levels. The current study was designed to investigate the effects of combinations of different environmental stresses on the transcriptome level of Arabidopsis genome using public microarray databases. We investigated the role of cyclopentenones in mediating plant responses to environmental stress through TGA (TGACG motif-binding factor) transcription factor, independently from jasmonic acid. Candidate genes were identified by comparing plants inoculated with Botrytis cinerea or treated with heat, salt or osmotic stress with non-inoculated or non-treated tissues. About 2.5% heat-, 19% salinity- and 41% osmotic stress-induced genes were commonly upregulated by B. cinerea-treatment; and 7.6%, 19% and 48% of genes were commonly downregulated by B. cinerea-treatment, respectively. Our results indicate that plant responses to biotic and abiotic stresses are mediated by several common regulatory genes. Comparisons between transcriptome data from Arabidopsis stressed-plants support our hypothesis that some molecular and biological processes involved in biotic and abiotic stress response are conserved. Thirteen of the common regulated genes to abiotic and biotic stresses were studied in detail to determine their role in plant resistance to B. cinerea. Moreover, a T-DNA insertion mutant of the Responsive to Dehydration gene (rd20), encoding for a member of the caleosin (lipid surface protein) family, showed an enhanced sensitivity to B. cinerea infection and drought. Overall, the overlapping of plant responses to abiotic and biotic stresses, coupled with the sensitivity of the rd20 mutant, may provide new interesting programs for increased plant resistance to multiple environmental stresses, and ultimately increases its chances to survive. Future research directions towards a

  5. The Epiphytic Fungus Pseudozyma aphidis Induces Jasmonic Acid- and Salicylic Acid/Nonexpressor of PR1-Independent Local and Systemic Resistance1[C][W

    PubMed Central

    Buxdorf, Kobi; Rahat, Ido; Gafni, Aviva; Levy, Maggie

    2013-01-01

    Pseudozyma spp. are yeast-like fungi, classified in the Ustilaginales, which are mostly epiphytic or saprophytic and are not pathogenic to plants. Several Pseudozyma species have been reported to exhibit biological activity against powdery mildews. However, previous studies have reported that Pseudozyma aphidis, which can colonize plant surfaces, is not associated with the ‎‎collapse of powdery ‎mildew colonies. In this report, we describe a novel P. aphidis strain and study its interactions with its plant host and the plant pathogen Botrytis cinerea. This isolate was found to secrete extracellular metabolites that inhibit various fungal pathogens in vitro and significantly reduce B. cinerea infection in vivo. Moreover, P. aphidis sensitized Arabidopsis (Arabidopsis thaliana) plants’ defense machinery via local and systemic induction of PATHOGENESIS-RELATED1 (PR1) and PLANT DEFENSIN1.2 (PDF1.2) expression. P. aphidis also reduced B. cinerea infection, locally and systemically, in Arabidopsis mutants impaired in jasmonic acid (JA) or salicylic acid (SA) signaling. Thus, in addition to direct inhibition, P. aphidis may inhibit B. cinerea infection via induced resistance in a manner independent of SA, JA, and Nonexpressor of PR1 (NPR1). P. aphidis primed the plant defense machinery and induced stronger activation of PDF1.2 after B. cinerea infection. Finally, P. aphidis fully or partially reconstituted PR1 and PDF1.2 expression in npr1-1 mutant and in plants with the SA hydroxylase NahG transgene, but not in a jasmonate resistant1-1 mutant, after B. cinerea infection, suggesting that P. aphidis can bypass the SA/NPR1, but not JA, pathway to activate PR genes. Thus, either partial gene activation is sufficient to induce resistance, or the resistance is not directed solely through PR1 and PDF1.2 but probably through other pathogen-resistance genes or pathways as well. PMID:23388119

  6. The epiphytic fungus Pseudozyma aphidis induces jasmonic acid- and salicylic acid/nonexpressor of PR1-independent local and systemic resistance.

    PubMed

    Buxdorf, Kobi; Rahat, Ido; Gafni, Aviva; Levy, Maggie

    2013-04-01

    Pseudozyma spp. are yeast-like fungi, classified in the Ustilaginales, which are mostly epiphytic or saprophytic and are not pathogenic to plants. Several Pseudozyma species have been reported to exhibit biological activity against powdery mildews. However, previous studies have reported that Pseudozyma aphidis, which can colonize plant surfaces, is not associated with the collapse of powdery mildew colonies. In this report, we describe a novel P. aphidis strain and study its interactions with its plant host and the plant pathogen Botrytis cinerea. This isolate was found to secrete extracellular metabolites that inhibit various fungal pathogens in vitro and significantly reduce B. cinerea infection in vivo. Moreover, P. aphidis sensitized Arabidopsis (Arabidopsis thaliana) plants' defense machinery via local and systemic induction of pathogenesis-related1 (PR1) and plant defensin1.2 (PDF1.2) expression. P. aphidis also reduced B. cinerea infection, locally and systemically, in Arabidopsis mutants impaired in jasmonic acid (JA) or salicylic acid (SA) signaling. Thus, in addition to direct inhibition, P. aphidis may inhibit B. cinerea infection via induced resistance in a manner independent of SA, JA, and Nonexpressor of PR1 (NPR1). P. aphidis primed the plant defense machinery and induced stronger activation of PDF1.2 after B. cinerea infection. Finally, P. aphidis fully or partially reconstituted PR1 and PDF1.2 expression in npr1-1 mutant and in plants with the SA hydroxylase NahG transgene, but not in a jasmonate resistant1-1 mutant, after B. cinerea infection, suggesting that P. aphidis can bypass the SA/NPR1, but not JA, pathway to activate PR genes. Thus, either partial gene activation is sufficient to induce resistance, or the resistance is not directed solely through PR1 and PDF1.2 but probably through other pathogen-resistance genes or pathways as well. PMID:23388119

  7. Two new antimicrobial metabolites from the endophytic fungus, Seimatosporium sp.

    PubMed

    Hussain, Hidayat; Krohn, Karsten; Schulz, Barbara; Draeger, Siegfried; Nazir, Mamona; Saleem, Muhammad

    2012-03-01

    Two new acaranoic acids, named seimatoporic acid A and B (1, and 2), together with six known compounds, R-(-)-mellein (3), cis-4-hydroxymellein (4), trans-4-hydroxymellein (5), 4R-hydroxy-5-methylmellein (6), (-)-5-hydroxymethylmellein (7), and ergosterol (8) were isolated from an endophytic fungus, Seimatosporium sp, by a bioassay-guided procedure. The structures of the new compounds have been assigned from analysis of the 1H and 13C NMR spectra, DEPT, and by 2D COSY, HMQC, HMBC and NOESY experiments. A mixture of compounds 1 and 2 showed strong antifungal activity against Botrytis cinerea, Septoria tritici, and Pyricularia oryzae. PMID:22545398

  8. Isolation and characterization of phosmidosine. A new antifungal nucleotide antibiotic.

    PubMed

    Uramoto, M; Kim, C J; Shin-Ya, K; Kusakabe, H; Isono, K; Phillips, D R; McCloskey, J A

    1991-04-01

    A new nucleotide antibiotic, phosmidosine was isolated from a culture filtrate of a newly isolated streptomycete identified as Streptomyces sp. RK-16. HRFAB-MS and elemental analysis established the molecular formula of C16H24N7O8P. 1H, 13C and 31P NMR indicated the presence of a methyl phosphate group and UV spectra were similar to those of 8-hydroxyadenosine. The antibiotic inhibited spore formation of Botrytis cinerea at the concentration of 0.25 micrograms/ml. PMID:2032945

  9. Effect of gamma irradiation on quality of kiwifruit ( Actinidia deliciosa var. deliciosa cv. Hayward)

    NASA Astrophysics Data System (ADS)

    kim, Kyoung-Hee; Yook, Hong-Sun

    2009-06-01

    Ionizing radiation is able to inactivate the three pathogens of Botrytis cinerea, Diaporthe actinidiae, and Botryosphaeria dothidea in kiwifruit. Irradiated kiwifruits appeared softer compared to non-irradiated kiwifruits. The color and organic acid content of kiwifruits were minimally affected by the irradiation. Irradiated fruits showed a decrease in the total soluble solids content with increasing irradiation dose. Irradiation of kiwifruit up to 3 kGy had negative effects on vitamin C content and antioxidant activity, but it contributed to improving sensory quality.

  10. Cerinolactone, a hydroxy-lactone derivative from Trichoderma cerinum.

    PubMed

    Vinale, Francesco; Girona, Isabel Arjona; Nigro, Marco; Mazzei, Pierluigi; Piccolo, Alessandro; Ruocco, Michelina; Woo, Sheridan; Rosa, David Ruano; Herrera, Carlos López; Lorito, Matteo

    2012-01-27

    A novel metabolite, 3-hydroxy-5-(6-isopropyl-3-methylene-3,4,4a,5,6,7,8,8a-octahydronaphthalen-2-yl)dihydrofuran-2-one, trivially named cerinolactone (1), has been isolated from culture filtrates of Trichoderma cerinum together with three known butenolides containing the 3,4-dialkylfuran-2(5H)-one nucleus, harzianolide (2), T39butenolide (3), and dehydroharzianolide (4). The structure of 1 was determined by spectroscopic methods, including UV, MS, and 1D and 2D NMR analyses. In vitro tests with the purified compound exhibited activity against Pythium ultimum, Rhizoctonia solani, and Botrytis cinerea. PMID:22196692

  11. Bacillus sp. BS061 Suppresses Powdery Mildew and Gray Mold

    PubMed Central

    Kim, Young-Sook; Song, Ja-Gyeong; Lee, In-Kyoung; Yeo, Woon-Hyung

    2013-01-01

    The use of a microorganism, or its secretions, to prevent plant disease offers an attractive alternative or supplement to synthetic fungicides for the management of plant disease without the negative effects of chemical control mechanisms. During a screening for microorganisms with the potential to be used as microbial fungicides, Bacillus sp. BS061 was isolated from a plant leaf. The strain BS061 potently inhibited the mycelial growth of Botrytis cinerea, and significantly reduced disease incidence of powdery mildew in cucumber and strawberry. We also found that the culture filtrate of BS061 inhibited the mycelial growth of various plant pathogens. PMID:23874134

  12. Antimicrobial activity of secondary metabolites from Streptomyces sp. K15, an endophyte in Houttuynia cordata Thunb.

    PubMed

    Chen, Huabao; Yang, Chunping; Ke, Tao; Zhou, Miaomiao; Li, Zhaojun; Zhang, Min; Gong, Guoshu; Hou, Taiping

    2015-01-01

    We isolated Streptomyces sp. K15 from the root tissue of Houttuynia cordata Thunb and found that some of its secondary metabolites exhibited significant antimicrobial activity against Botrytis cinerea. Moreover, we separated, purified and identified the major active ingredient to be 2-pyrrol formic acid by using silica gel column chromatography, high-performance liquid chromatography and NMR analysis of the spectral data. 2-Pyrrol formic acid critically inhibited the growth of some phytopathogenic bacteria. Therefore, it has potential value in agricultural applications. PMID:25675117

  13. Bioactivity guided isolation of antifungal compounds from the liverwort Bazzania trilobata (L.) S.F. Gray.

    PubMed

    Scher, Jochen M; Speakman, John-Bryan; Zapp, Josef; Becker, Hans

    2004-09-01

    A dichloromethane and a methanol extract of the liverwort Bazzania trilobata showed antifungal activity against the phytopathogenic fungi Botrytis cinerea, Cladosporium cucumerinum, Phythophthora infestans, Pyricularia oryzae and Septoria tritici. Bioautography on thin-layer chromatograms was used to isolate six antifungal sesquiterpenes: 5- and 7-hydroxycalamenene, drimenol, drimenal, viridiflorol, gymnomitrol and three bisbibenzyls: 6 ',8'-dichloroisoplagiochin C, isoplagiochin D and 6'-chloroisoplagiochin D. Furthermore we report the isolation of gymnomitr-8(12)-en-4-one and the new coumarin 7,8-dihydroxycoumarin-7-O-beta-D-glucuronide. Their structures have been elucidated based on extensive NMR spectral evidence. PMID:15451321

  14. Synthesis, Antifungal Activities and Qualitative Structure Activity Relationship of Carabrone Hydrazone Derivatives as Potential Antifungal Agents

    PubMed Central

    Wang, Hao; Ren, Shuang-Xi; He, Ze-Yu; Wang, De-Long; Yan, Xiao-Nan; Feng, Jun-Tao; Zhang, Xing

    2014-01-01

    Aimed at developing novel fungicides for relieving the ever-increasing pressure of agricultural production caused by phytopathogenic fungi, 28 new hydrazone derivatives of carabrone, a natural bioactive sesquisterpene, in three types were designed, synthesized and their antifungal activities against Botrytis cinerea and Colletotrichum lagenarium were evaluated. The result revealed that all the derivatives synthesized exhibited considerable antifungal activities in vitro and in vivo, which led to the improved activities for carabrone and its analogues and further confirmed their potential as antifungal agents. PMID:24619221

  15. Use of a non-homologous end-joining-deficient strain (delta-ku70) of the biocontrol fungus Trichoderma virens to investigate the function of the laccase gene lcc1 in sclerotia degradation

    PubMed Central

    Catalano, Valentina; Vergara, Mariarosaria; Hauzenberger, Jasmin R.; Seiboth, Bernhard; Sarrocco, Sabrina; Vannacci, Giovanni; Kubicek, Christian P.

    2010-01-01

    The aim of this study was to apply a generated Δtku70 strain with increased homologous recombination efficiency from the mycoparasitic fungus Trichoderma virens for studying the involvement of laccases in the degradation of sclerotia of plant pathogenic fungi. Inactivation of the non-homologous end-joining pathway has become a successful tool in filamentous fungi to overcome poor targeting efficiencies for genetic engineering. Here, we applied this principle to the biocontrol fungus T. virens, strain I10, by deleting its tku70 gene. This strain was subsequently used to delete the laccase gene lcc1, which we found to be expressed after interaction of T. virens with sclerotia of the plant pathogenic fungi Botrytis cinerea and Sclerotinia sclerotiorum. Lcc1 was strongly upregulated at early colonization of B. cinerea sclerotia and steadily induced during colonization of S. sclerotiorum sclerotia. The Δtku70Δlcc1 mutant was altered in its ability to degrade the sclerotia of B. cinerea and S. sclerotiorum. Interestingly, while the decaying ability for B. cinerea sclerotia was significantly decreased, that to degrade S. sclerotiorum sclerotia was even enhanced, suggesting the operation of different mechanisms in the mycoparasitism of these two types of sclerotia by the laccase LCC1. PMID:20872221

  16. Loss of Ceramide Kinase in Arabidopsis Impairs Defenses and Promotes Ceramide Accumulation and Mitochondrial H2O2 Bursts[C][W

    PubMed Central

    Bi, Fang-Cheng; Liu, Zhe; Wu, Jian-Xin; Liang, Hua; Xi, Xue-Li; Fang, Ce; Sun, Tie-Jun; Yin, Jian; Dai, Guang-Yi; Rong, Chan; Greenberg, Jean T.; Su, Wei-Wei; Yao, Nan

    2014-01-01

    Arabidopsis thaliana plants that lack ceramide kinase, encoded by ACCELERATED CELL DEATH5 (ACD5), display spontaneous programmed cell death late in development and accumulate substrates of ACD5. Here, we compared ceramide accumulation kinetics, defense responses, ultrastructural features, and sites of reactive oxygen species (ROS) production in wild-type and acd5 plants during development and/or Botrytis cinerea infection. Quantitative sphingolipid profiling indicated that ceramide accumulation in acd5 paralleled the appearance of spontaneous cell death, and it was accompanied by autophagy and mitochondrial ROS accumulation. Plants lacking ACD5 differed significantly from the wild type in their responses to B. cinerea, showing earlier and higher increases in ceramides, greater disease, smaller cell wall appositions (papillae), reduced callose deposition and apoplastic ROS, and increased mitochondrial ROS. Together, these data show that ceramide kinase greatly affects sphingolipid metabolism and the site of ROS accumulation during development and infection, which likely explains the developmental and infection-related cell death phenotypes. The acd5 plants also showed an early defect in restricting B. cinerea germination and growth, which occurred prior to the onset of cell death. This early defect in B. cinerea restriction in acd5 points to a role for ceramide phosphate and/or the balance of ceramides in mediating early antifungal responses that are independent of cell death. PMID:25149397

  17. HISTONE MONOUBIQUITINATION1 Interacts with a Subunit of the Mediator Complex and Regulates Defense against Necrotrophic Fungal Pathogens in Arabidopsis[W

    PubMed Central

    Dhawan, Rahul; Luo, Hongli; Foerster, Andrea Maria; AbuQamar, Synan; Du, Hai-Ning; Briggs, Scott D.; Scheid, Ortrun Mittelsten; Mengiste, Tesfaye

    2009-01-01

    This work examines the role of the Arabidopsis thaliana RING E3 ligase, HISTONE MONOUBIQUITINATION1 (HUB1) in disease resistance. Loss-of-function alleles of HUB1 show increased susceptibility to the necrotrophic fungal pathogens Botrytis cinerea and Alternaria brassicicola, whereas HUB1 overexpression conferred resistance to B. cinerea. By contrast, responses to the bacterial pathogen Pseudomonas syringae are unaltered in hub1 plants. hub1 mutants have thinner cell walls but increased callose around an infection site. HUB1 acts independently of jasmonate, but ethylene (ET) responses and salicylate modulate the resistance of hub1 mutants to necrotrophic fungi. The ET response factor ETHYLENE INSENSITIVE2 is epistatic to HUB1 for A. brassicicola resistance but additive to HUB1 for B. cinerea resistance. HUB1 interacts with MED21, a subunit of the Arabidopsis Mediator, a conserved complex that regulates RNA polymerase II. RNA interference lines with reduced MED21 expression are highly susceptible to A. brassicicola and B. cinerea, whereas T-DNA insertion alleles are embryonic lethal, suggesting an essential role for MED21. However, HUB1-mediated histone H2B modification is independent of histone H3 and DNA methylation. In sum, histone H2B monoubiquitination is an important chromatin modification with regulatory roles in plant defense against necrotrophic fungi most likely through modulation of gene expression. PMID:19286969

  18. The microbiome of the leaf surface of Arabidopsis protects against a fungal pathogen.

    PubMed

    Ritpitakphong, Unyarat; Falquet, Laurent; Vimoltust, Artit; Berger, Antoine; Métraux, Jean-Pierre; L'Haridon, Floriane

    2016-05-01

    We have explored the importance of the phyllosphere microbiome in plant resistance in the cuticle mutants bdg (BODYGUARD) or lacs2.3 (LONG CHAIN FATTY ACID SYNTHASE 2) that are strongly resistant to the fungal pathogen Botrytis cinerea. The study includes infection of plants under sterile conditions, 16S ribosomal DNA sequencing of the phyllosphere microbiome, and isolation and high coverage sequencing of bacteria from the phyllosphere. When inoculated under sterile conditions bdg became as susceptible as wild-type (WT) plants whereas lacs2.3 mutants retained the resistance. Adding washes of its phyllosphere microbiome could restore the resistance of bdg mutants, whereas the resistance of lacs2.3 results from endogenous mechanisms. The phyllosphere microbiome showed distinct populations in WT plants compared to cuticle mutants. One species identified as Pseudomonas sp isolated from the microbiome of bdg provided resistance to B. cinerea on Arabidopsis thaliana as well as on apple fruits. No direct activity was observed against B. cinerea and the action of the bacterium required the plant. Thus, microbes present on the plant surface contribute to the resistance to B. cinerea. These results open new perspectives on the function of the leaf microbiome in the protection of plants. PMID:26725246

  19. Overexpression of AtSHN1/WIN1 Provokes Unique Defense Responses

    PubMed Central

    Sela, Dikla; Buxdorf, Kobi; Shi, Jian Xin; Feldmesser, Ester; Schreiber, Lukas; Aharoni, Asaph; Levy, Maggie

    2013-01-01

    The plant cell cuticle serves as the first barrier protecting plants from mechanical injury and invading pathogens. The cuticle can be breached by cutinase-producing pathogens and the degradation products may activate pathogenesis signals in the invading pathogens. Cuticle degradation products may also trigger the plant’s defense responses. Botrytis cinerea is an important plant pathogen, capable of attacking and causing disease in a wide range of plant species. Arabidopsis thaliana shn1-1D is a gain-of-function mutant, which has a modified cuticular lipid composition. We used this mutant to examine the effect of altering the whole-cuticle metabolic pathway on plant responses to B. cinerea attack. Following infection with B. cinerea, the shn1-1D mutant discolored more quickly, accumulated more H2O2, and showed accelerated cell death relative to wild-type (WT) plants. Whole transcriptome analysis of B. cinerea-inoculated shn1-1D vs. WT plants revealed marked upregulation of genes associated with senescence, oxidative stress and defense responses on the one hand, and genes involved in the magnitude of defense-response control on the other. We propose that altered cutin monomer content and composition of shn1-1D plants triggers excessive reactive oxygen species accumulation and release which leads to a strong, unique and uncontrollable defense response, resulting in plant sensitivity and death. PMID:23922943

  20. Evaluation of Lettuce Germplasm Resistance to Gray Mold Disease for Organic Cultivations

    PubMed Central

    Shim, Chang Ki; Kim, Min Jeong; Kim, Yong Ki; Jee, Hyeong Jin

    2014-01-01

    This study was conducted to evaluate the resistance of 212 accessions of lettuce germplasm to gray mold disease caused by Botrytis cinerea. The lettuce germplasm were composed of five species: Lactuca sativa (193 accessions), L. sativa var. longifolia (2 accessions), L. sativa var. crispa (2 accessions), L. saligna (2 accessions), and L. serriola (1 accession); majority of these originated from Korea, Netherlands, USA, Russia, and Bulgaria. After 35 days of spray inoculation with conidial suspension (3×107 conidia/ml) of B. cinerea on the surface of lettuce leaves, tested lettuce germplasm showed severe symptoms of gray mold disease. There were 208 susceptible accessions to B. cinerea counted with 100% of disease incidence and four resistant accessions, IT908801, K000598, K000599, and K021055. Two moderately resistant accessions of L. sativa, K021055 and IT908801, showed 20% of disease incidence of gray mold disease at 45 days after inoculation; and two accessions of L. saligna, K000598 and K000599, which are wild relatives of lettuce germplasm with loose-leaf type, showed complete resistance to B. cinerea. These four accessions are candidates for breeding lettuce cultivars resistant to gray mold disease. PMID:25288990

  1. Ruta montana L. leaf essential oil and extracts: characterization of bioactive compounds and suppression of crown gall disease

    PubMed Central

    Hammami, Inés; Smaoui, Slim; Hsouna, Anis Ben; Hamdi, Naceur; Triki, Mohamed Ali

    2015-01-01

    The aims of this study were to assess the antimicrobial efficacy of the leaf essential oil and the leaf extracts of R. montana against Botrytis cinerea, Fusarium oxysporum, Verticillium dahliae, Aspergillus oryzae and Fusarium solani. The oil (1.000 µg/disk) and the extracts (1.500 µg/disk) revealed a remarkable antifungal effect against the tested plant pathogenic fungi with a radial growth inhibition percentage of 40.0-80.0 % and 5.0-58.0 %, respectively along with their respective MIC values ranging from 100 to 1100 µg/mL and 250 to 3000 µg/mL. The oil had a strong detrimental effect on spore germination of all the tested plant pathogens along with the concentration as well as time-dependent kinetic inhibition of Fusarium oxysporum. Also, the oil exhibited a potent in vivo antifungal effect against Botrytis cinerea on tomato plants. Experiments carried out in plant revealed that the essential oil was slightly effective in suppression of gall formation induced by Agrobacterium tumefaciens on bitter almond. The results of this study indicate that the oil and extracts of R. montana leaves could become natural alternatives to synthetic fungicides to control certain important plant microbial diseases. The GC-MS analysis determined that 28 compounds, which represented 89.03 % of total oil, were present in the oil containing mainly 1-butene, methylcyclopropane, 2-butene and caryophyllene oxide. PMID:26417353

  2. Characterization of combined power plasma jet using AC high voltage and nanosecond pulse for reactive species composition control

    NASA Astrophysics Data System (ADS)

    Takashima, Keisuke; Konishi, Hideaki; Kato, Toshiaki; Kaneko, Toshiro

    2014-10-01

    In the application studies for both bio-medical and agricultural applications, the roles of the reactive oxide and/or nitride species generated in the plasma has been reported as a key to control the effects and ill-effects on the living organism. The correlation between total OH radical exposure from an air atmospheric pressure plasma jet and the sterilization threshold on Botrytis cinerea is presented. With the increase of the OH radical exposure to the Botrytis cinerea, the probability of sterilization is increased. In this study, to resolve the roles of reactive species including OH radicals, a combined power plasma jet using nanosecond pulses and low-frequency sinusoidal AC high voltage (a few kHz) is studied for controlling the composition of the reactive species. The nanosecond pulses are superimposed on the AC voltage which is in synchronization with the AC phase. The undergoing work to characterize the combined power discharge with electric charge and voltage cycle on the plasma jet will also be presented to discuss the discharge characteristics to control the composition of the reactive species.

  3. Lipoxygenase activity and sanguinarine production in cell suspension cultures of California poppy (Eschscholtzia californica CHAM.).

    PubMed

    Kollárová, R; Oblozinský, M; Kováciková, V; Holková, I; Balazová, A; Pekárová, M; Hoffman, P; Bezáková, L

    2014-08-01

    In this study we investigated the influence of biotic elicitor (phytopathogenic fungus Botrytis cinerea) and abiotic elicitors (methyljasmonate [MJ] and salicylic acid [SA]) on lipoxygenase (LOX) activity and sanguinarine production in cell suspension cultures of California poppy (Eschscholtzia californica CHAM.). We have observed different time effects of elicitors (10, 24, 48 and 72 h) on LOX activity and production of sanguinarine in in vitro cultures. All elicitors used in the experiments evidently increased the LOX activity and sanguinarine production in contrast to control samples. The highest LOX activities were determined in samples elicitated by MJ after 48 h and 72 h and the lowest LOX activities (in contrast to control samples) were detected after biotic elicitation by Botrytis cinerea. These activities showed about 50% lower level against the activities after MJ elicitation. The maximal amount of sanguinarine was observed after 48 h in MJ treated cultures (429.91 mg/g DCW) in comparision with control samples. Although all elicitors affect the sanguinarine production, effect of SA and biotic elicitor on sanguinarine accumulation in in vitrocultures was not so significant than after MJ elicitation. PMID:25158577

  4. Ruta montana L. leaf essential oil and extracts: characterization of bioactive compounds and suppression of crown gall disease.

    PubMed

    Hammami, Inés; Smaoui, Slim; Hsouna, Anis Ben; Hamdi, Naceur; Triki, Mohamed Ali

    2015-01-01

    The aims of this study were to assess the antimicrobial efficacy of the leaf essential oil and the leaf extracts of R. montana against Botrytis cinerea, Fusarium oxysporum, Verticillium dahliae, Aspergillus oryzae and Fusarium solani. The oil (1.000 µg/disk) and the extracts (1.500 µg/disk) revealed a remarkable antifungal effect against the tested plant pathogenic fungi with a radial growth inhibition percentage of 40.0-80.0 % and 5.0-58.0 %, respectively along with their respective MIC values ranging from 100 to 1100 µg/mL and 250 to 3000 µg/mL. The oil had a strong detrimental effect on spore germination of all the tested plant pathogens along with the concentration as well as time-dependent kinetic inhibition of Fusarium oxysporum. Also, the oil exhibited a potent in vivo antifungal effect against Botrytis cinerea on tomato plants. Experiments carried out in plant revealed that the essential oil was slightly effective in suppression of gall formation induced by Agrobacterium tumefaciens on bitter almond. The results of this study indicate that the oil and extracts of R. montana leaves could become natural alternatives to synthetic fungicides to control certain important plant microbial diseases. The GC-MS analysis determined that 28 compounds, which represented 89.03 % of total oil, were present in the oil containing mainly 1-butene, methylcyclopropane, 2-butene and caryophyllene oxide. PMID:26417353

  5. Endophytic fungus strain 28 isolated from Houttuynia cordata possesses wide-spectrum antifungal activity.

    PubMed

    Pan, Feng; Liu, Zheng-Qiong; Chen, Que; Xu, Ying-Wen; Hou, Kai; Wu, Wei

    2016-01-01

    The aim of this paper is to identify and investigate an endophytic fungus (strain 28) that was isolated from Houttuynia cordata Thunb, a famous and widely-used Traditional Chinese Medicine. Based on morphological methods and a phylogenetic analysis of ITS sequences, this strain was identified as Chaetomium globosum. An antifungal activity bioassay demonstrated that the crude ethyl acetate (EtOAc) extracts of strain 28 had a wide antifungal spectrum and strong antimicrobial activity, particularly against Exserohilum turcicum (Pass.) Leonard et Suggs, Botrytis cinerea persoon and Botrytis cinerea Pers. ex Fr. Furthermore, the fermentation conditions, extraction method and the heat stability of antifungal substances from strain 28 were also studied. The results showed that optimal antifungal activity can be obtained with the following parameters: using potato dextrose broth (PDB) as the base culture medium, fermentation for 4-8d (initial pH: 7.5), followed by extraction with EtOAc. The extract was stable at temperatures up to 80°C. This is the first report on the isolation of endophytic C. globosum from H. cordata to identify potential alternative biocontrol agents that could provide new opportunities for practical applications involving H. cordata. PMID:26991297

  6. Effect of prior drought and pathogen stress on Arabidopsis transcriptome changes to caterpillar herbivory.

    PubMed

    Davila Olivas, Nelson H; Coolen, Silvia; Huang, Pingping; Severing, Edouard; van Verk, Marcel C; Hickman, Richard; Wittenberg, Alexander H J; de Vos, Martin; Prins, Marcel; van Loon, Joop J A; Aarts, Mark G M; van Wees, Saskia C M; Pieterse, Corné M J; Dicke, Marcel

    2016-06-01

    In nature, plants are exposed to biotic and abiotic stresses that often occur simultaneously. Therefore, plant responses to combinations of stresses are most representative of how plants respond to stresses. We used RNAseq to assess temporal changes in the transcriptome of Arabidopsis thaliana to herbivory by Pieris rapae caterpillars, either alone or in combination with prior exposure to drought or infection with the necrotrophic fungus Botrytis cinerea. Pre-exposure to drought stress or Botrytis infection resulted in a significantly different timing of the caterpillar-induced transcriptional changes. Additionally, the combination of drought and P. rapae induced an extensive downregulation of A. thaliana genes involved in defence against pathogens. Despite a more substantial growth reduction observed for plants exposed to drought plus P. rapae feeding compared with P. rapae feeding alone, this did not affect weight increase of this specialist caterpillar. Plants respond to combined stresses with phenotypic and transcriptional changes that differ from the single stress situation. The effect of a previous exposure to drought or B. cinerea infection on transcriptional changes to caterpillars is largely overridden by the stress imposed by caterpillars, indicating that plants shift their response to the most recent stress applied. PMID:26847575

  7. Antimicrobial action effect and stability of nanosized silica hybrid Ag complex.

    PubMed

    Kim, Hwa-Jung; Park, Hae-Jun; Choi, Seong-Ho

    2011-07-01

    Nanosized silica hybrid silver complex (NSS) showing strong antifungal activity, in which nanosilver (nano-Ag) was bound to silica (SiO2) molecules, was synthesized via gamma-irradiation at room temperature. NSS was characterized via field emission scanning electron microscopy (FESEM), energy-dispersive X-ray spectroscopy (EDXS), ultraviolet-visible (UV-Vis) spectrophotometer, and thermogravimetric analysis (TGA). The FESEM images and EDXS data showed that well-dispersed 3-to-10-nm Ag nanoparticles (core part) were loaded onto the outer parts of 5-to-20 nm SiO2 nanoparticles. The antifungal efficiency of NSS was evaluated against Rhizoctonia solani, Botrytis cinerea, and Colletotrichum gloeosporioides. In the case of Rhizoctonia solani, the growth rate was decreased typically by more than 90% at a 6 microg/ml concentration of NSS as a medium additive. The antifungal-action mechanism was investigated via transmission electron microscopy (TEM) analysis of the NSS treatment against Botrytis cinerea. The stability and antimicrobial activity of NSS were determined, using the plate culture method, from several water samples containing NSS after 7-day NSS treatment. Moreover, the NSS solution maintained stable antifungal activity for at least 24 mos. These results suggest that NSS, an environment-friendly nanomaterial, can be used as strongly effective growth inhibitor of various microorganisms, making it applicable to diverse antimicrobial-control systems. PMID:22121607

  8. Fungal ABC transporters and microbial interactions in natural environments.

    PubMed

    Schoonbeek, Henk-jan; Raaijmakers, Jos M; De Waard, Maarten A

    2002-11-01

    In natural environments, microorganisms are exposed to a wide variety of antibiotic compounds produced by competing organisms. Target organisms have evolved various mechanisms of natural resistance to these metabolites. In this study, the role of ATP-binding cassette (ABC) transporters in interactions between the plant-pathogenic fungus Botrytis cinerea and antibiotic-producing Pseudomonas bacteria was investigated in detail. We discovered that 2,4-diacetylphloroglucinol, phenazine-1-carboxylic acid and phenazine-1-carboxamide (PCN), broad-spectrum antibiotics produced by Pseudomonas spp., induced expression of several ABC transporter genes in B. cinerea. Phenazines strongly induced expression of BcatrB, and deltaBcatrB mutants were significantly more sensitive to these antibiotics than their parental strain. Treatment of B. cinerea germlings with PCN strongly affected the accumulation of [14C]fludioxonil, a phenylpyrrole fungicide known to be transported by BcatrB, indicating that phenazines also are transported by BcatrB. Pseudomonas strains producing phenazines displayed a stronger antagonistic activity in vitro toward ABcatrB mutants than to the parental B. cinerea strain. On tomato leaves, phenazine-producing Pseudomonas strains were significantly more effective in reducing gray mold symptoms incited by a ABcatrB mutant than by the parental strain. We conclude that the ABC transporter BcatrB provides protection to B. cinerea in phenazine-mediated interactions with Pseudomonas spp. Collectively, these results indicate that fungal ABC transporters can play an important role in antibiotic-mediated interactions between bacteria and fungi in plant-associated environments. The implications of these findings for the implementation and sustainability of crop protection by antagonistic microorganisms are discussed. PMID:12423022

  9. Discrimination of fungal infections on grape berries via spectral signatures

    NASA Astrophysics Data System (ADS)

    Molitor, Daniel; Griesser, Michaela; Schütz, Erich; Khuen, Marie-Therese; Schefbeck, Christa; Ronellenfitsch, Franz Kai; Schlerf, Martin; Beyer, Marco; Schoedl-Hummel, Katharina; Anhalt, Ulrike; Forneck, Astrid

    2016-04-01

    The fungal pathogens Botrytis cinerea and Penicillium expansum are causing economic damages on grapevine worldwide. Especially the simultaneous occurrence of both often results in off-flavours highly threatening wine quality. For the classification of grape quality as well as for the determination of targeted enological treatments, the knowledge of the level of fungal attack is of highest interest. However, visual assessment and pathogen discrimination are cost-intensive. Consequently, a pilot laboratory study aimed at (i) detecting differences in spectral signatures between grape berry lots with different levels of infected berries (B. cinerea and/or P. expansum) and (ii) detecting links between spectral signatures and biochemical as well as quantitative molecular markers for fungal attack. To this end, defined percentages (infection levels) of table grape berries were inoculated with fungal spore suspensions. Spectral measurements were taken using a FieldSpec 3 Max spectroradiometer (ASD Inc., Boulder/Colorado, USA) in regular intervals after inoculation. In addition, fungal attack was determined enzymatically) and quantitatively (real-time PCR). In addition, gluconic acid concentrations (as a potential markers for fungal attack) were determined photometrically. Results indicate that based on spectral signatures, a discrimination of P. expansum and B. cinerea infections as well as of different B. cinerea infection levels is possible. Real-time PCR analyses, detecting DNA levels of both fungi, showed yet a low detection level. Whereas the gluconic acid concentrations turned out to be specific for the two fungi tested (B. cinerea vs. P. expansum) and thus may serve as a differentiating biochemical marker. Correlation analyses between spectral measurements and biological data (gluconic acid concentrations, fungi DNA) as well as further common field and laboratory trials are targeted.

  10. Reliability of the entomovector technology using Prestop-Mix and Bombus terrestris L. as a fungal disease biocontrol method in open field.

    PubMed

    Karise, Reet; Dreyersdorff, Gerit; Jahani, Mona; Veromann, Eve; Runno-Paurson, Eve; Kaart, Tanel; Smagghe, Guy; Mänd, Marika

    2016-01-01

    Botrytis cinerea Pers.:Fr. is a major plant pathogen, and a new approach is needed for its control in strawberry to minimise the increasing use of synthetic fungicides. The biofungicide Prestop-Mix, which contains Gliocladium catenulatum, is effective against Botrytis infections; however, the need for frequent applications increases the costs for farmers. Here, we demonstrate that bumble bees, Bombus terrestris L., effectively disseminate the preparation onto flowers in open field conditions. Over the course of three years, we found a highly significant decrease in the rate of Botrytis infection. Pathogen control was achieved with relatively low numbers of G. catenulatum spores per flower, even using flowers that are not highly attractive to bumble bees. An even distribution of spores was detected up to 100 m from the hives, either due to primary inoculation by bumble bees or secondary distribution by other flower visitors such as honey bees and solitary bees. We showed that the application of a biocontrol agent by bumble bees is reliable for the use of environmentally friendly pest control strategies in northern climatic conditions. This low cost technology is especially relevant for organic farming. This study provides valuable information for introducing this method into practice in open strawberry fields. PMID:27530075

  11. Reliability of the entomovector technology using Prestop-Mix and Bombus terrestris L. as a fungal disease biocontrol method in open field

    PubMed Central

    Karise, Reet; Dreyersdorff, Gerit; Jahani, Mona; Veromann, Eve; Runno-Paurson, Eve; Kaart, Tanel; Smagghe, Guy; Mänd, Marika

    2016-01-01

    Botrytis cinerea Pers.:Fr. is a major plant pathogen, and a new approach is needed for its control in strawberry to minimise the increasing use of synthetic fungicides. The biofungicide Prestop-Mix, which contains Gliocladium catenulatum, is effective against Botrytis infections; however, the need for frequent applications increases the costs for farmers. Here, we demonstrate that bumble bees, Bombus terrestris L., effectively disseminate the preparation onto flowers in open field conditions. Over the course of three years, we found a highly significant decrease in the rate of Botrytis infection. Pathogen control was achieved with relatively low numbers of G. catenulatum spores per flower, even using flowers that are not highly attractive to bumble bees. An even distribution of spores was detected up to 100 m from the hives, either due to primary inoculation by bumble bees or secondary distribution by other flower visitors such as honey bees and solitary bees. We showed that the application of a biocontrol agent by bumble bees is reliable for the use of environmentally friendly pest control strategies in northern climatic conditions. This low cost technology is especially relevant for organic farming. This study provides valuable information for introducing this method into practice in open strawberry fields. PMID:27530075

  12. Ecofriendly hot water treatment reduces postharvest decay and elicits defense response in kiwifruit.

    PubMed

    Chen, Huizhen; Cheng, Zhe; Wisniewski, Michael; Liu, Yongsheng; Liu, Jia

    2015-10-01

    Hot water treatment (HWT) of fruit is an effective approach for managing postharvest decay of fruits and vegetables. In the present study, the effects of HWT (45 °C for 10 min) on the growth of Botrytis cinerea and Penicillium expansum in vitro, and gray (B. cinerea) and blue mold (P. expansum) development in kiwifruit were investigated. HWT effectively inhibited spore germination and germ tube elongation of B. cinerea and P. expansum. Reactive oxygen species accumulation and protein impairment in the fungi triggered by HWT contributed to the inhibitory effect. Results of in vivo studies showed that HWT controlled gray and blue mold in kiwifruit stored at 4 and 25 °C. HWT induced a significant increase in the activity of antioxidant enzymes, including catalase and peroxidase, and the level of total phenolic compounds in kiwifruit. These findings indicate that the inhibition of postharvest decay in kiwifruit by HWT is associated with the inhibition of spore germination of both fungal pathogens and the elicitation of defense response in the kiwifruit host. Moreover, HWT used in this study did not impair fruit quality. HWT appears to represent a potential non-chemical alternative for the effective management of postharvest decay of kiwifruit. PMID:26002370

  13. Effect of Thymol and Linalool Fumigation on Postharvest Diseases of Table Grapes

    PubMed Central

    Shin, Mi Ho; Kim, Jin-Hee; Choi, Hyo-Won; Keum, Yoong Soo

    2014-01-01

    Several postharvest diseases of table grapes (Vitis vinifera) occur during storage, and gray mold rot is a particularly severe disease because the causal agent, Botrytis cinerea, grows at temperatures as low as 0℃. Other postharvest diseases, such as those caused by Penicillium spp. and Aspergillus spp., also often lead to deterioration in the quality of table grapes after harvest. The use of plant essential oils such as thymol and linalool, to reduce postharvest diseases in several kinds of fruits, including table grapes and oranges, has received much attention in European countries. However, to the best of our knowledge there has been no report of the use of thymol fumigation to control gray mold in table grapes in Korea. Thymol (30 µg/mL) and linalool (120 µg/mL) significantly inhibited mycelial growth and conidia germination of B. cinerea. The occurrence rate of gray mold rot of B. cinerea and other unknown fungi was significantly reduced by fumigation with 30 µg/mL thymol in several table grape cultivars, such as Campbell early, Muscat Bailey A, Sheridan, and Geobong. In this study, fumigation with 30 µg/mL thymol, had no influence on the sugar content and hardness of grapes, but reduced fungal infection significantly. This suggests that 30 µg/mL thymol could be utilized to reduce deterioration of grapes due to gray mold and other fungal infections during long-term storage. PMID:25346603

  14. Pectin Biosynthesis Is Critical for Cell Wall Integrity and Immunity in Arabidopsis thaliana.

    PubMed

    Bethke, Gerit; Thao, Amanda; Xiong, Guangyan; Li, Baohua; Soltis, Nicole E; Hatsugai, Noriyuki; Hillmer, Rachel A; Katagiri, Fumiaki; Kliebenstein, Daniel J; Pauly, Markus; Glazebrook, Jane

    2016-02-01

    Plant cell walls are important barriers against microbial pathogens. Cell walls of Arabidopsis thaliana leaves contain three major types of polysaccharides: cellulose, various hemicelluloses, and pectins. UDP-d-galacturonic acid, the key building block of pectins, is produced from the precursor UDP-d-glucuronic acid by the action of glucuronate 4-epimerases (GAEs). Pseudomonas syringae pv maculicola ES4326 (Pma ES4326) repressed expression of GAE1 and GAE6 in Arabidopsis, and immunity to Pma ES4326 was compromised in gae6 and gae1 gae6 mutant plants. These plants had brittle leaves and cell walls of leaves had less galacturonic acid. Resistance to specific Botrytis cinerea isolates was also compromised in gae1 gae6 double mutant plants. Although oligogalacturonide (OG)-induced immune signaling was unaltered in gae1 gae6 mutant plants, immune signaling induced by a commercial pectinase, macerozyme, was reduced. Macerozyme treatment or infection with B. cinerea released less soluble uronic acid, likely reflecting fewer OGs, from gae1 gae6 cell walls than from wild-type Col-0. Although both OGs and macerozyme-induced immunity to B. cinerea in Col-0, only OGs also induced immunity in gae1 gae6. Pectin is thus an important contributor to plant immunity, and this is due at least in part to the induction of immune responses by soluble pectin, likely OGs, that are released during plant-pathogen interactions. PMID:26813622

  15. Higher Potassium Concentration in Shoots Reduces Gray Mold in Sweet Basil.

    PubMed

    Yermiyahu, Uri; Israeli, Lior; David, Dalia Rav; Faingold, Inna; Elad, Yigal

    2015-08-01

    Nutritional elements can affect plant susceptibility to plant pathogens, including Botrytis cinerea. We tested the effect of potassium (K) fertilization on gray mold in sweet basil grown in pots, containers, and soil. Increased K in the irrigation water and in the sweet basil tissue resulted in an exponential decrease in gray mold severity. Potassium supplied to plants by foliar application resulted in a significant decrease in gray mold in plants grown with a low rate of K fertigation. Lower K fertigation resulted in a significant increase in B. cinerea infection under semi-commercial conditions. Gray mold severity in harvested shoots was significantly negatively correlated with K concentration in the irrigation solution, revealing resistance to B. cinerea infection as a result of high K concentration in sweet basil tissue. Gray mold was reduced following K foliar application of the plants. In general, there was no synergy between the fertigation and foliar spray treatments. Proper K fertilization can replace some of the required chemical fungicide treatments and it may be integrated into gray mold management for improved disease suppression. PMID:25760521

  16. Effect of thymol and linalool fumigation on postharvest diseases of table grapes.

    PubMed

    Shin, Mi Ho; Kim, Jin-Hee; Choi, Hyo-Won; Keum, Yoong Soo; Chun, Se Chul

    2014-09-01

    Several postharvest diseases of table grapes (Vitis vinifera) occur during storage, and gray mold rot is a particularly severe disease because the causal agent, Botrytis cinerea, grows at temperatures as low as 0℃. Other postharvest diseases, such as those caused by Penicillium spp. and Aspergillus spp., also often lead to deterioration in the quality of table grapes after harvest. The use of plant essential oils such as thymol and linalool, to reduce postharvest diseases in several kinds of fruits, including table grapes and oranges, has received much attention in European countries. However, to the best of our knowledge there has been no report of the use of thymol fumigation to control gray mold in table grapes in Korea. Thymol (30 µg/mL) and linalool (120 µg/mL) significantly inhibited mycelial growth and conidia germination of B. cinerea. The occurrence rate of gray mold rot of B. cinerea and other unknown fungi was significantly reduced by fumigation with 30 µg/mL thymol in several table grape cultivars, such as Campbell early, Muscat Bailey A, Sheridan, and Geobong. In this study, fumigation with 30 µg/mL thymol, had no influence on the sugar content and hardness of grapes, but reduced fungal infection significantly. This suggests that 30 µg/mL thymol could be utilized to reduce deterioration of grapes due to gray mold and other fungal infections during long-term storage. PMID:25346603

  17. Effect of Cinnamic Acid for Controlling Gray Mold on Table Grape and Its Possible Mechanisms of Action.

    PubMed

    Zhang, Zhanquan; Qin, Guozheng; Li, Boqiang; Tian, Shiping

    2015-09-01

    Cinnamic acid (CA) is an organic acid and is widely used in food industry as a common food additive. Previous studies showed that CA has the antimicrobial activity in vitro, but little is known about the effect of CA on controlling the fruit decay in vivo. In present study, we showed that application of CA was significantly effective on controlling the gray mold of table grape caused by Botrytis cinerea. CA can directly inhibit the mycelial growth of B. cinerea on potato dextrose agar plates. The mechanisms by which CA inhibited fungal growth were assayed by staining the spores with fluorescent dyes propidium iodide and 7-dichlorodihydrofluorescein diacetate, respectively. The results indicated that CA can damage the integrity of plasma membrane and induce the intracellular reactive oxygen species level of B. cinerea which were responsible for the reduction of growth rate. Meanwhile, CA treatment significantly stimulated the activities of peroxidase and polyphenol oxidase which were closely related to the resistance of plant. Taken together, this study suggested that CA was effective on controlling the gray mold of table grape in postharvest period by inhibiting the growth of pathogen and inducing the resistance of host. PMID:26143055

  18. Enzyme activity and effect of heat treatment on some fungal diseases of postharvest tomato fruits.

    PubMed

    Amer, M A; El-Abd, S M; Mansour, F G F

    2013-01-01

    The activity of heat treatments for controlling tomato black mould caused by Alternaria alternata (Fr.) Keissler and grey mould caused by Botrytis cinerea Pers. Ex. Pers. wWas tested. Spore suspension of the grey mold pathogen which was exposed to hot water treatment at 58 degrees C for 6 min., failed to germinate, whereas A. alternata failed to germinate when exposed to hot water at 58 degrees C, for 8 min. Exposing discs bearing growth of any both tested fungi to hot air at 48 degrees C for 72 h caused growth suppression for the both tested pathogens. Dipping tomato fruits in hot water at 58 degrees C for 8 min or keeping in hot air for 72 h at 40 degrees C prevented decay development in non inoculated or artificially inoculated fruits with A. alternata and B. cinerea up to 12 days when stored at 24 degrees C. The high amount of polygalacturonase (PG) and pectinmethylestrase (BME) enzymes were investigated in infected fruits by the two tested pathogens as compared by healthy ones. The culture filtrate of A. alternata had the highest protease activity, white B. cinerea was the lowest. Protease activity was higher when CD liquid medium contain casein was used. PMID:25151835

  19. The rhizobacterium Arthrobacter agilis produces dimethylhexadecylamine, a compound that inhibits growth of phytopathogenic fungi in vitro.

    PubMed

    Velázquez-Becerra, Crisanto; Macías-Rodríguez, Lourdes I; López-Bucio, José; Flores-Cortez, Idolina; Santoyo, Gustavo; Hernández-Soberano, Christian; Valencia-Cantero, Eduardo

    2013-12-01

    Plant diseases caused by fungal pathogens such as Botrytis cinerea and the oomycete Phytophthora cinnamomi affect agricultural production worldwide. Control of these pests can be done by the use of fungicides such as captan, which may have deleterious effects on human health. This study demonstrates that the rhizobacterium Arthrobacter agilis UMCV2 produces volatile organic compounds that inhibit the growth of B. cinerea in vitro. A single compound from the volatile blends, namely dimethylhexadecylamine (DMHDA), could inhibit the growth of both B. cinerea and P. cinnamomi when supplied to the growth medium in low concentrations. DMHDA also inhibited the growth of beneficial fungi Trichoderma virens and Trichoderma atroviride but at much higher concentrations. DMHDA-related aminolipids containing 4, 8, 10, 12, and 14 carbons in the alkyl chain were tested for their inhibitory effect on the growth of the pathogens. The results show that the most active compound from those tested was dimethyldodecylamine. This effect correlates with a decrease in the number of membrane lipids present in the mycelium of the pathogen including eicosanoic acid, (Z)-9-hexadecenoic acid, methyl ester, and (Z)-9-octadecenoic acid, methyl ester. Strawberry leaflets treated with DMHDA were not injured by the compound. These data indicate that DMHDA and related compounds, which can be produced by microorganisms may effectively inhibit the proliferation of certain plant pathogens. PMID:23674267

  20. Activation of Plant Innate Immunity by Extracellular High Mobility Group Box 3 and Its Inhibition by Salicylic Acid

    PubMed Central

    Choi, Hyong Woo; Manohar, Murli; Manosalva, Patricia; Tian, Miaoying; Moreau, Magali; Klessig, Daniel F.

    2016-01-01

    Damage-associated molecular pattern molecules (DAMPs) signal the presence of tissue damage to induce immune responses in plants and animals. Here, we report that High Mobility Group Box 3 (HMGB3) is a novel plant DAMP. Extracellular HMGB3, through receptor-like kinases BAK1 and BKK1, induced hallmark innate immune responses, including i) MAPK activation, ii) defense-related gene expression, iii) callose deposition, and iv) enhanced resistance to Botrytis cinerea. Infection by necrotrophic B. cinerea released HMGB3 into the extracellular space (apoplast). Silencing HMGBs enhanced susceptibility to B. cinerea, while HMGB3 injection into apoplast restored resistance. Like its human counterpart, HMGB3 binds salicylic acid (SA), which results in inhibition of its DAMP activity. An SA-binding site mutant of HMGB3 retained its DAMP activity, which was no longer inhibited by SA, consistent with its reduced SA-binding activity. These results provide cross-kingdom evidence that HMGB proteins function as DAMPs and that SA is their conserved inhibitor. PMID:27007252

  1. In vitro and in vivo evaluation of the antioxidant and prooxidant activity of phenolic compounds obtained from grape (Vitis vinifera) pomace.

    PubMed

    Cotoras, Milena; Vivanco, Herman; Melo, Ricardo; Aguirre, María; Silva, Evelyn; Mendoza, Leonora

    2014-01-01

    The antioxidant and/or prooxidant ability of extracts obtained from wine waste were analyzed using in vitro and in vivo assays. Cyclic voltammetry was used as the in vitro assay to determine the antioxidant and/or prooxidant properties and, the in vivo effect on mycelial growth of the fungus Botrytis cinerea was evaluated. In addition, the prooxidant activity was evaluated by intracellular oxidation of compound 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) in B. cinerea. The extracts used in this study were obtained from grape pomace of Cabernet Sauvignon, Carménère and Syrah varieties from the Misiones de Rengo Vineyard by simple extraction, using methanol/HCl 1% (v/v), ethanol 70% (v/v), or Soxhlet extraction. According to the results obtained, gallic acid was the most represented phenolic compound independent of grape variety and extraction method. In addition, vanillic acid; protocatechuic acid, syringic acid, quercetin and kaempferol were found in the extracts. From this study it was possible concluded that, depending of the method of extraction of the grape residues and the grape variety (Cabernet Sauvignon, Carménère and Syrah), the extracts showed antioxidant and/or prooxidant activity. However, no correlation can be established between the anodic oxidation potentials of the extracts and their effect on the fungus B. cinerea. PMID:25521116

  2. ARACINs, Brassicaceae-Specific Peptides Exhibiting Antifungal Activities against Necrotrophic Pathogens in Arabidopsis1[OPEN

    PubMed Central

    Neukermans, Jenny; Inzé, Annelies; Mathys, Janick; De Coninck, Barbara; van de Cotte, Brigitte; Cammue, Bruno P.A.; Van Breusegem, Frank

    2015-01-01

    Plants have developed a variety of mechanisms to cope with abiotic and biotic stresses. In a previous subcellular localization study of hydrogen peroxide-responsive proteins, two peptides with an unknown function (designated ARACIN1 and ARACIN2) have been identified. These peptides are structurally very similar but are transcriptionally differentially regulated during abiotic stresses during Botrytis cinerea infection or after benzothiadiazole and methyl jasmonate treatments. In Arabidopsis (Arabidopsis thaliana), these paralogous genes are positioned in tandem within a cluster of pathogen defense-related genes. Both ARACINs are small, cationic, and hydrophobic peptides, known characteristics for antimicrobial peptides. Their genes are expressed in peripheral cell layers prone to pathogen entry and are lineage specific to the Brassicaceae family. In vitro bioassays demonstrated that both ARACIN peptides have a direct antifungal effect against the agronomically and economically important necrotrophic fungi B. cinerea, Alternaria brassicicola, Fusarium graminearum, and Sclerotinia sclerotiorum and yeast (Saccharomyces cerevisiae). In addition, transgenic Arabidopsis plants that ectopically express ARACIN1 are protected better against infections with both B. cinerea and A. brassicicola. Therefore, we can conclude that both ARACINs act as antimicrobial peptides. PMID:25593351

  3. Identification of reference genes suitable for qRT-PCR in grapevine and application for the study of the expression of genes involved in pterostilbene synthesis.

    PubMed

    Gamm, Magdalena; Héloir, Marie-Claire; Kelloniemi, Jani; Poinssot, Benoît; Wendehenne, David; Adrian, Marielle

    2011-04-01

    The recent publication of the grapevine genome sequence facilitates the use of qRT-PCR to study gene expression changes. For this approach, reference genes are commonly used to normalize data and their stability of expression should be systematically validated. Among grapevine defenses is the production of the antimicrobial stilbenic phytoalexins, notably the highly fungitoxic pterostilbene, which plays a crucial role in grapevine interaction with Plasmopara viticola and Botrytis cinerea. As a resveratrol O-methyltransferase (ROMT) gene involved in pterostilbene synthesis was recently identified, we investigated the accumulation of the corresponding transcripts to those of two other stilbene biosynthesis related genes phenylalanine ammonia lyase (PAL) and stilbene synthase (STS) in response to pathogen infection. Using three computer-based statistical methods and C(t) values or LRE method generated values as input data, we have first identified two reference genes (VATP16 and 60SRP) suitable for normalization of qPCR expression data obtained in grapevine leaves and berries infected by P. viticola and B. cinerea, respectively. Next, we have highlighted that the expression of ROMT is induced in P. viticola-infected leaves and also in B. cinerea-infected berries, confirming the involvement of pterostilbene in grapevine defenses. PMID:21340517

  4. A dual role for plant quinone reductases in host-fungus interaction.

    PubMed

    Heyno, Eiri; Alkan, Noam; Fluhr, Robert

    2013-11-01

    Quinone reductases (QR, EC 1.5.6.2) are flavoproteins that protect organisms from oxidative stress. The function of plant QRs has not as yet been addressed in vivo despite biochemical evidence for their involvement in redox reactions. Here, using knock-out (KO) and overexpressing lines, we studied the protective role of two groups of Arabidopsis thaliana cytosolic QRs, Nqr (NAD(P)H:quinone oxidoreductase) and Fqr (flavodoxin-like quinone reductase), in response to infection by necrotrophic fungi. The KO lines nqr(-) and fqr1(-) displayed significantly slower development of lesions of Botrytis cinerea and Sclerotinia sclerotium in comparison to the wild type (WT). Consistent with this observation, the overexpressing line FQR1(+) was hypersensitive to the pathogens. Both the nqr(-) and fqr1(-) displayed increased fluorescence of 2',7'-dichlorofluorescein,‬ a reporter for reactive oxygen species in response to B. cinerea. Infection by B. cinerea was accompanied with increased Nqr and Fqr1 protein levels in the WT as revealed by western blotting. In addition, a marked stimulation of salicylic acid-sensitive transcripts and suppression of jasmonate-sensitive transcripts was observed in moderately wounded QR KO mutant leaves, a condition mimicking the early stage of infection. In contrast to the above observations, germination of conidia was accelerated on leaves of QR KO mutants in comparison with the WT and FQR1(+). The same effect was observed in water-soluble leaf surface extracts. It is proposed that the altered interaction between B. cinerea and the QR mutants is a consequence of subtly altered redox state of the host, which perturbs host gene expression in response to environmental stress such as fungal growth.‬‬‬‬‬‬ PMID:23464356

  5. Antifungal Substances from Streptomyces sp. A3265 Antagonistic to Plant Pathogenic Fungi

    PubMed Central

    Van Minh, Nguyen; Woo, E-Eum; Kim, Ji-Yul; Kim, Dae-Won; Hwang, Byung Soon; Lee, Yoon-Ju; Lee, In-Kyoung

    2015-01-01

    In a previous study, we identified a Streptomyces sp., A3265, as exhibiting potent antifungal activity against various plant pathogenic fungi, including Botrytis cinerea, Colletotrichum gloeosporioides, and Rhizoctonia solani. This strain also exhibited a biocontrolling effect against ginseng root rot and damping-off disease, common diseases of ginseng and other crops. In this study, we isolated two antifungal substances responsible for this biocontrolling effect via Diaion HP-20 and Sephadex LH-20 column chromatography, medium pressure liquid chromatography, and high-performance liquid chromatography. These compounds were identified as guanidylfungin A and methyl guanidylfungin A by spectroscopic methods. These compounds exhibited potent antimicrobial activity against various plant pathogenic fungi as well as against bacteria. PMID:26539051

  6. Burdock fructooligosaccharide induces fungal resistance in postharvest Kyoho grapes by activating the salicylic acid-dependent pathway and inhibiting browning.

    PubMed

    Sun, Fei; Zhang, Pengying; Guo, Moran; Yu, Wenqian; Chen, Kaoshan

    2013-05-01

    Burdock fructooligosaccharide (BFO) is a natural elicitor from Arcitum lappa. The effects of BFO in controlling postharvest disease in grape, apple, banana, kiwi, citrus, strawberry, and pear were investigated. The disease index, decay percentage, and area under the disease progress curve indicated that BFO has general control effects on postharvest disease of fruits. Kyoho grapes were studied to elucidate the mechanism of BFO in boosting the resistance of grapes to Botrytis cinerea infection. BFO treatment induced upregulation of the npr1, pr1, pal, and sts genes, and inhibited the total phenol content decrease, which activated chitinase and β-1,3-glucanase. These results indicated that the salicylic acid-dependent signalling pathway was induced. The delayed colour change and peroxidase and polyphenoloxidase activity suggested that BFO delayed grape browning. The reduced respiration rate, weight loss, and titratable acidity prolonged the shelf life of postharvest grapes. BFO is a promising elicitor in postharvest disease control. PMID:23265522

  7. Chemical characterization and antifungal activity of essential oil of capitula from wild Indian Tagetes patula L.

    PubMed

    Romagnoli, C; Bruni, R; Andreotti, E; Rai, M K; Vicentini, C B; Mares, D

    2005-04-01

    The essential oil extracted by steam distillation from the capitula of Indian Tagetes patula, Asteraceae, was evaluated for its antifungal properties and analyzed by gas chromatography and gas chromatography-mass spectrometry. Thirty compounds were identified, representing 89.1% of the total detected. The main components were piperitone (24.74%), piperitenone (22.93%), terpinolene (7.8%), dihydro tagetone (4.91%), cis-tagetone (4.62%), limonene (4.52%), and allo-ocimene (3.66%). The oil exerted a good antifungal activity against two phytopathogenic fungi, Botrytis cinerea and Penicillium digitatum, providing complete growth inhibition at 10 microl/ml and 1.25 microl/ml, respectively. The contribution of the two main compounds, piperitone and piperitenone, to the antifungal efficacy was also evaluated and ultrastructural modifications in mycelia were observed via electron microscopy, evidencing large alterations in hyphal morphology and a multisite mechanism of action. PMID:15868213

  8. Thyme essential oil as a defense inducer of tomato against gray mold and Fusarium wilt.

    PubMed

    Ben-Jabeur, Maissa; Ghabri, Emna; Myriam, Machraoui; Hamada, Walid

    2015-09-01

    The potential of thyme essential oil in controlling gray mold and Fusarium wilt and inducing systemic acquired resistance in tomato seedlings and tomato grown in hydroponic system was evaluated. Thyme oil highly reduced 64% of Botrytis cinerea colonization on pretreated detached leaves compared to untreated control. Also, it played a significant decrease in Fusarium wilt severity especially at7 days post treatment when it was reduced to 30.76%. To explore the plant pathways triggered in response to thyme oil, phenolic compounds accumulation and peroxidase activity was investigated. Plant response was observed either after foliar spray or root feeding in hydroponics which was mostly attributed to peroxidases accumulation rather than phenolic compounds accumulation, and thyme oil seems to be more effective when applied to the roots. PMID:26002413

  9. Design, Synthesis, and Fungicidal Activities of Novel 5-Methyl-1H-1,2,3- trizole-4-carboxyl Amide Analogues.

    PubMed

    Wang, Zhen-Jun; Yang, Hui-Hui; Tian, Lei; Zhao, Wei-Guang

    2016-01-01

    Succinate dehydrogenase inhibitors (SDHIs) are fungicides with an amide bond widely used to control plant diseases caused by phytopathogenic fungi. Because of broad spectrum activity of new SDHIs, they have attracted wide attention from the research community. A series of structurally novel SDHIs with a bioactive 1,2,3-triazole moiety were designed and synthesized. Bioactivity screening showed that some of designed N-phenethyl-1,2,3-trizole-4-carboxyl amide analogues exhibited good fungicidal activities toward Sclerotinia sclerotiorum and Botrytis cinerea, while some of Nbenzyl- 1,2,3-trizole-4-carboxyl amide analogues exhibited good fungicidal activities toward Phytophthora capsici and Cercospora arachidicola. EC50 value of compound 5d against Cercospora arachidicola (6.6 µg/mL) was lower than that of chlorothalonil (12.3 µg/mL). PMID:26558376

  10. Alocasin, an anti-fungal protein from rhizomes of the giant taro Alocasia macrorrhiza.

    PubMed

    Wang, H X; Ng, T B

    2003-03-01

    An anti-fungal protein designated alocasin was isolated from the rhizomes of the giant taro Alocasia macrorrhiza. The isolation protocol involved ion exchange chromatography on diethylaminoethyl (DEAE)-cellulose, ion exchange chromatography on sulfopropyl (SP)-Sepharose, and gel filtration on Superdex 75. Alocasin, which was unadsorbed on DEAE-cellulose and SP-Sepharose, possessed the N-terminal sequence APEGEV, which exhibited some similarity to that of the miraculin-like anti-fungal protein from Pisum sativum legumes. It demonstrated a molecular mass of 11kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, and displayed anti-fungal activity against Botrytis cinerea. Alocasin reduced the activity of HIV-1 reverse transcriptase. It exhibited weak hemagglutinating activity, only at a concentration of 1mg/ml. PMID:12651101

  11. Microwave Assistant Synthesis, Antifungal Activity and DFT Theoretical Study of Some Novel 1,2,4-Triazole Derivatives Containing Pyridine Moiety

    PubMed Central

    Sun, Guo-Xiang; Yang, Ming-Yan; Shi, Yan-Xia; Sun, Zhao-Hui; Liu, Xing-Hai; Wu, Hong-Ke; Li, Bao-Ju; Zhang, Yong-Gang

    2014-01-01

    In order to investigate the biological activity of novel 1,2,4-triazole compounds, seventeen novel 1,2,4-triazole derivatives containing pyridine moiety were synthesized under microwave assistant condition by multi-step reactions. The structures were characterized by 1H NMR, MS and elemental analyses. The target compounds were evaluated for their fungicidal activities against Stemphylium lycopersici (Enjoji) Yamamoto, Fusarium oxysporum. sp. cucumebrium, and Botrytis cinerea in vivo, and the results indicated that some of the title compounds displayed excellent fungicidal activities. Theoretical calculation of the title compound was carried out with B3LYP/6-31G (d,p). The full geometry optimization was carried out using 6-31G (d,p) basis set, and the frontier orbital energy, atomic net charges were discussed, and the structure-activity relationship was also studied. PMID:24815069

  12. Development of a Selective Medium for the Fungal Pathogen Cylindrocarpon destructans Using Radicicol.

    PubMed

    Kang, Yunhee; Lee, Seung-Ho; Lee, Jungkwan

    2014-12-01

    The soil-borne ascomycete fungus Cylindrocarpon destructans causes ginseng root rot disease and produces various secondary metabolites such as brefeldin A and radicicol. The slow growth of this fungus compared with other plant pathogenic and saprophytic fungi in soil disturbs isolation of this fungus from soil and infected ginseng. In this study, we developed a selective medium for C. destructans using radicicol produced by this fungus. Supplementing 50 mg/L of radicicol to medium inhibited the mycelia growth of other fungi including Botrytis cinerea, Rhizoctonia solani and Alternaria panax, but did not affect the growth of C. destructans. In addition, conidia germination of other fungal species except for C. destructans was inhibited in submerged culture supplemented with radicicol. This medium provides a very efficient tool for isolating C. destructans and also can be used as an enrichment medium for this fungus. PMID:25506308

  13. Development of a Selective Medium for the Fungal Pathogen Cylindrocarpon destructans Using Radicicol

    PubMed Central

    Kang, Yunhee; Lee, Seung-Ho; Lee, Jungkwan

    2014-01-01

    The soil-borne ascomycete fungus Cylindrocarpon destructans causes ginseng root rot disease and produces various secondary metabolites such as brefeldin A and radicicol. The slow growth of this fungus compared with other plant pathogenic and saprophytic fungi in soil disturbs isolation of this fungus from soil and infected ginseng. In this study, we developed a selective medium for C. destructans using radicicol produced by this fungus. Supplementing 50 mg/L of radicicol to medium inhibited the mycelia growth of other fungi including Botrytis cinerea, Rhizoctonia solani and Alternaria panax, but did not affect the growth of C. destructans. In addition, conidia germination of other fungal species except for C. destructans was inhibited in submerged culture supplemented with radicicol. This medium provides a very efficient tool for isolating C. destructans and also can be used as an enrichment medium for this fungus. PMID:25506308

  14. [Antibiotic activity of some fungi].

    PubMed

    Savchuk, Ia I; Tsyganenko, K S; Zaĭchenko, O M

    2013-01-01

    Biological activity of pure extracts of cultural filtrates of Aspergillus niveus 2411, Myrothecium cinctum 910, Ulocladium consortiale 960, Penicillium sp. 10-51 concerning wide spectrum of test-organisms was investigated. It was shown that the extracts had high levels of antibacterial activity against Gram-positive microorganisms, especially against Bacillus genus. But their activity against Gram-negative bacteria was a bit lower. On the other hand, metabolites of M. cinctum 910 and Penicillium sp. 10-51 did show the activity concerning phytopathogenic bacteria. Extracts of fungi showed fungistatic activity against yeasts, but they were not so active concerning fungal test-cultures. Extracts of A. niveus 2411, Penicillium sp. 10-51 suppressed the growth of Phoma betae. The highest level of fungistatic activity was shown by metabolites of M. cinctum 910. They showed activity against Aspergillus genus strains and phytopathogenic isolates of Fusarium lactis, Rhizoctonia solani and Botrytis cinerea. PMID:24479314

  15. Synthesis, Characterization, and Antifungal Activity of Phenylpyrrole-Substituted Tetramic Acids Bearing Carbonates.

    PubMed

    Xu, Wen-Qin; Chen, Min; Wang, Kun-Yao; Ren, Zheng-Jiao; Lu, Ai-Min; Yang, Chun-Long

    2016-01-01

    For the aim of discovering new fungicide, a series of phenylpyrrole-substituted tetramic acid derivatives bearing carbonates 6a-q were designed and synthesized via 4-(2,4-dioxopyrrolidin-3-ylidene)-4-(phenylamino)butanoic acids 4a-k and the cyclized products 1',3,4,5'-tetrahydro-[2,3'-bipyrrolylidene]-2',4',5(1H)-triones 5a-k. The compounds were characterized using IR, ¹H- and (13)C-NMR spectroscopy, mass spectrometry (EI-MS), and elemental analysis. The structure of 6b was confirmed by X-ray diffraction crystallography. The title compounds 6a-q were bioassayed in vitro against the phytopathogenic fungi Fusarium graminearum, Botrytis cinerea and Rhizoctonia solani at a concentration of 100 μg/mL, respectively. Most compounds displayed good inhibitory activity. PMID:27007370

  16. Bioactive metabolites from the mycelia of the basidiomycete Hericium erinaceum.

    PubMed

    Lu, Qiang-Qiang; Tian, Jun-Mian; Wei, Jing; Gao, Jin-Ming

    2014-01-01

    Seven known compounds, three diketopiperazine alkaloids, 12β-hydroxyverruculogen TR-2 (1), fumitremorgin C (2) and methylthiogliotoxin (5), two hetero-spirocyclic γ-lactam alkaloids, pseurotin A (3) and FD-838 (4), and cerevisterol (6) and herierin IV (7), were isolated from the mycelia of the basidiomycete Hericium erinaceum and identified by spectroscopic analyses. The antioxidant and antifungal activities of compounds 1-6 were evaluated. The results indicated that compounds 1, 3 and 6 exhibited potential antioxidant activity against DPPH (2, 2-diphenyl-1-picrylhydrazyl) radical with their IC50 data of ca. 12 μM, compared with positive control tertiary butylhydroquinone. In addition, compound 4 significantly inhibited the growth of two plant fungal pathogens Botrytis cinerea and Glomerella cingulata with an minimum inhibitory concentration of 6.25 μM for each, similar to that of the positive fungicide, carbendazim. Compounds 1-5 were isolated from the genus Hericium for the first time. PMID:24635196

  17. Isolation and antifungal activity of kakuol, a propiophenone derivative from Asarum sieboldii rhizome.

    PubMed

    Lee, Jung Yeop; Moon, Surk Sik; Hwang, Byung Kook

    2005-08-01

    An antifungal substance active against Colletotrichum orbiculare (Berk & Mont) Arx was isolated from the methanol extracts of Asarum sieboldii (Miq) Maek rhizomes. High-resolution MS, NMR and UV spectral data confirmed that the antifungal substance is kakuol, 2-hydroxy-4,5-methylenedioxypropiophenone. Colletotrichum orbiculare was most sensitive to kakuol, with MIC of 10 microg ml(-1). Kakuol also completely inhibited the mycelial growth of Botrytis cinerea Pers ex Fr and Cladosporium cucumerinum Ellis & Arthur at 50 microg ml(-1) and 30 microg ml(-1), respectively. However, no antimicrobial activity was found against yeast and bacteria even at 100 microg ml(-1). Kakuol exhibited a protective activity against the development of anthracnose disease on cucumber plants. The control efficacy of kakuol against the anthracnose disease was in general somewhat less than that of the commercial fungicide chlorothalonil. This is the first report to demonstrate in vitro and in vivo antifungal activity of kakuol against C. orbiculare infection. PMID:15846774

  18. Production of a newly isolated Paenibacillus polymyxa biocontrol agent using monosodium glutamate wastewater and potato wastewater.

    PubMed

    Gu, Likun; Bai, Zhihui; Jin, Bo; Zhang, Jianyun; Li, Wenying; Zhuang, Guoqiang; Zhang, Hongxun

    2010-01-01

    A phyllosphere bacterial strain EBL-06 was isolated from wheat leaves. The morphology, cultural characteristics, phospholipid fatty acids, physiological and antagonistic fungus activities of this strain were investigated. A phylogenetic tree was constructed by comparing with the published 16S rDNA sequences of the relevant bacteria. The results showed that the isolate EBL-06 was a strain of Paenibacillus polymyxa; this strain performed a high level of antagonistic fungus activity toward a broad spectrum of phytopathogens, such as Botrytis cinerea, Cladosporium cucumerinum, Fusarium spp. The isolate EBL-06 can grow well using monosodium glutamate wastewater (MGW) and potato wastewater (PW) as culture medium. The maximum yield of 6.5 x 10(9) CFU/mL of the isolate EBL-06 anti-fungus biocontrol agent was reached in 15 hr cultivation at 28 degrees C, pH 6.0-7.5 using the mixture of MGW and PW (1:9). PMID:21174972

  19. Cloning and overexpression of antifungal barley chitinase gene in Escherichia coli.

    PubMed

    Kirubakaran, S Isaac; Sakthivel, N

    2007-03-01

    Plant chitinases are pathogenesis-related proteins, which are believed to be involved in plant defense responses to pathogen infection. In this study, chitinase gene from barley was cloned and overexpressed in Escherichia coli. Chitinase (35 kDa) was isolated and purified. Since the protein was produced as insoluble inclusion bodies, the protein was solubilized and refolded. Purified chitinase exerted broad-spectrum antifungal activity against Botrytis cinerea (blight of tobacco), Pestalotia theae (leaf spot of tea), Bipolaris oryzae (brown spot of rice), Alternaria sp. (grain discoloration of rice), Curvularia lunata (leaf spot of clover) and Rhizoctonia solani (sheath blight of rice). Due to the potential of broad-spectrum antifungal activity barley chitinase gene can be used to enhance fungal-resistance in crop plants such as rice, tobacco, tea and clover. PMID:17029984

  20. Synthetic Ultrashort Cationic Lipopeptides Induce Systemic Plant Defense Responses against Bacterial and Fungal Pathogens ▿

    PubMed Central

    Brotman, Yariv; Makovitzki, Arik; Shai, Yechiel; Chet, Ilan; Viterbo, Ada

    2009-01-01

    A new family of synthetic, membrane-active, ultrashort lipopeptides composed of only four amino acids linked to fatty acids was tested for the ability to induce systemic resistance and defense responses in plants. We found that two peptides wherein the third residue is a d-enantiomer (italic), C16-KKKK and C16-KLLK, can induce medium alkalinization of tobacco suspension-cultured cells and expression of defense-related genes in cucumber and Arabidopsis seedlings. Moreover, these compounds can prime systemic induction of antimicrobial compounds in cucumber leaves similarly to the plant-beneficial fungus Trichoderma asperellum T203 and provide systemic protection against the phytopathogens Botrytis cinerea B05, Pseudomonas syringae pv. lachrimans, and P. syringae pv. tomato DC3000. Thus, short cationic lipopeptides are a new category of compounds with potentially high utility in the induction of systemic resistance in plants. PMID:19542326

  1. Vulgarinin, a broad-spectrum antifungal peptide from haricot beans (Phaseolus vulgaris).

    PubMed

    Wong, Jack Ho; Ng, Tzi Bun

    2005-08-01

    From the seeds of haricot beans (Phaseolus vulgaris), an antifungal peptide with a molecular mass around 7 kDa was purified by using a simple protocol consisting of affinity chromatography on Affi-gel blue gel and gel filtration on Superdex 75. This peptide named vulgarinin manifested an antifungal activity toward fungal species such as Fusarium oxysporum, Mycosphaerella arachidicola, Physalospora piricola and Botrytis cinerea, and an antibacterial action on Mycobacterium phlei, Bacillus megaterium, Bacillus subtilis and Proteus vulgaris. It also inhibited proliferation in leukemia cell lines L1210 and M1 and breast cancer cell line MCF-7. This peptide could reduce the activity of HIV-1 reverse transcriptase and inhibited translation in a cell-free rabbit reticulocyte lysate system. Its antifungal activity was retained after incubation with trypsin. PMID:15896669

  2. Sesquin, a potent defensin-like antimicrobial peptide from ground beans with inhibitory activities toward tumor cells and HIV-1 reverse transcriptase.

    PubMed

    Wong, Jack Ho; Ng, Tzi Bun

    2005-07-01

    An antifungal peptide with a molecular mass around 7 kDa and an N-terminal sequence highly homologous to defensin was isolated from ground beans (Vigna sesquipedalis cv. 'Ground Bean'). The peptide was adsorbed on Affi-gel blue gel and on Mono S. It exerted an antifungal action on Botrytis cinerea, Fusarium oxysporum and Mycosphaerella arachidicola; and an antibacterial action on Escherichia coli B, Proteus vulgaris, Mycobacterium phlei and Bacillus megaterium. The antimicrobial activity was inhibited in presence of the 5 mM CaCl2 and MgCl2, but no inhibition was observed in 5 mM NaCl. The peptide exerted antiproliferative activity toward breast cancer (MCF-7) cells and leukemia M1 cells, this activity could not be inhibited by the ions mentioned above. It also exhibited some inhibitory activity toward human immunodeficiency virus-type 1 reverse transcriptase. PMID:15949629

  3. Lunatusin, a trypsin-stable antimicrobial peptide from lima beans (Phaseolus lunatus L.).

    PubMed

    Wong, Jack Ho; Ng, Tzi Bun

    2005-11-01

    An anti-fungal peptide designated as lunatusin, with a molecular mass around 7kDa, was purified from the seeds of Chinese lima bean (Phaseolus lunatus L.). The peptide was isolated using a simple protocol consisting of affinity chromatography on Affi-gel blue gel and gel filtration on Superdex 75. Lunatusin exerted an anti-fungal activity toward fungal species such as Fusarium oxysporum, Mycosphaerella arachidicola and Botrytis cinerea, and an antibacterial action on, Bacillus megaterium, Bacillus subtilis, Proteus vulgaris and Mycobacterium phlei. It also inhibited proliferation in the breast cancer cell line MCF-7. Lunatusin reduced the activity of HIV-1 reverse transcriptase and it also inhibited translation in a cell-free rabbit reticulocyte lysate system. Its anti-fungal activity was retained after incubation with trypsin. Lunatusin elicited a mitogenic response from mouse splenocytes. PMID:16269344

  4. Triterpenoids from the Herbs of Salicornia bigelovii.

    PubMed

    Shan, Yu; Li, Huan; Guan, Fuqin; Chen, Yu; Yin, Min; Wang, Ming; Feng, Xu; Wang, Qizhi

    2015-01-01

    A new nortriterpene saponin, 3-O-β-d-glucuronopyranosyl-30-norolean-12,20(29)-dien-23- oxo-28-oic acid, namely bigelovii D (11), was isolated from the hydroalcoholic extract of herbs of Salicornia bigelovii along with 10 known saponins (1-10). Their chemical structures were identified on the basis of spectroscopic analyses including two-dimensional NMR and a comparison with literature data. Some of these compounds showed potent antifungal activities in vitro. Compounds 3, 4, 5, 6, 7, 10 and 11 demonstrated potent inhibitory activities against Colletotrichum gloeosporioides and compound 11 displayed broad-spectrum inhibitory activity against Alternaria alternata, A. solani, Botrytis cinerea, C. gloeosporioides, Fusarium graminearum, F. verticilloides, Thanatephorus cucumeris and Sclerotinia sclerotiorum, with EC50 values ranging from 13.6 to 36.3 μg/mL. PMID:26569214

  5. Antifungal and antibacterial metabolites from an endophytic Aspergillus sp. associated with Melia azedarach.

    PubMed

    Xiao, Jian; Zhang, Qiang; Gao, Yu-Qi; Shi, Xin-Wei; Gao, Jin-Ming

    2014-01-01

    Seven known metabolites, dianhydro-aurasperone C (1), isoaurasperone A (2), fonsecinone A (3), asperpyrone A (4), asperazine (5), rubrofusarin B (6) and (R)-3-hydroxybutanonitrile (7), were isolated from the culture of Aspergillus sp. KJ-9, a fungal endophyte isolated from Melia azedarach and identified by spectroscopic methods. All isolates were evaluated in vitro against several phytopathogenic fungi (Gibberella saubinetti, Magnaporthe grisea, Botrytis cinerea, Colletotrichum gloeosporioides and Alternaria solani) and pathogenic bacteria (Escherichia coli, Bacillus subtilis, Staphyloccocus aureus and Bacillus cereus). Compounds 3 and 7 were active against almost all phytopathogenic fungi tested with minimum inhibitory concentration (MIC) range of 6.25-50 μM. Moreover, compound 3 was active against all pathogenic bacteria with MIC in the range of 25-100 μM. Compound 7 is a rare new natural product isolated from a natural source for the first time, and the detailed NMR data of 1 were first assigned. PMID:24708541

  6. Is it possible to increase the aloin content of Aloe vera by the use of ultraviolet light?

    PubMed

    Martínez-Romero, Domingo; Guillén, Fabián; Pérez-Aguilar, Henoc; Castillo, Salvador; Serrano, María; Zapata, Pedro J; Valero, Daniel

    2013-03-01

    In this paper, the effects of ultraviolet (UV) treatments on the aloin content of Aloe vera L. gel have been analyzed. UV-A treatment to A. vera plants for 36 days led to an increase in the aloin concentration in gel, rind tissue, and latex, while a decrease in chlorophylls a and b occurred in the photosynthetic tissue as a consequence of UV treatment. The growth of Penicillium digitatum and Botrytis cinerea (artificially inoculated on the leaf surface) was drastically decreased in UV-A-treated leaves, which could be attributed to the increase in the aloin concentration by the UV-A treatment. In addition, UV-C treatment to detached leaves also led to an increase in the gel aloin concentration, at higher levels than occurred with UV-A treatment, although leaves showed severe lesions after 48 h of treatment. PMID:23406252

  7. Optimization for rapid synthesis of silver nanoparticles and its effect on phytopathogenic fungi

    NASA Astrophysics Data System (ADS)

    Krishnaraj, C.; Ramachandran, R.; Mohan, K.; Kalaichelvan, P. T.

    In this present study, silver nanoparticles were synthesized by green chemistry approach using Acalypha indica leaf extract as reducing agents. The reaction medium employed in the synthesis process was optimized to attain better yield, controlled size and stability. Further, the biosynthesized silver nanoparticles were conformed through UV-vis spectrum, XRD and HR-TEM analyses. Different concentration of silver nanoparticles were tested to know the inhibitory effect of fungal plant pathogens namely Alternaria alternata, Sclerotinia sclerotiorum, Macrophomina phaseolina, Rhizoctonia solani, Botrytis cinerea and Curvularia lunata. Interestingly, 15 mg concentration of silver nanoparticles showed excellent inhibitory activity against all the tested pathogens. Thus, the obtained results clearly suggest that silver nanoparticles may have important applications in controlling various plant diseases caused by fungi.

  8. Cryopreservation of the Pinewood Nematode, Bursaphelenchus spp.

    PubMed

    Riga, E; Webster, J M

    1991-10-01

    Populations of three isolates of Bursaphelenchus xylophilus, the pinewood nematode, and one of B. mucronatus were treated with three cryoprotectants at -70 C for 24 hours followed by deep freezing at -180 C in liquid nitrogen for different periods of time. A solution of 15% glycerol, 35% buffer S, and 50% M9, or 1% aqueous solution of dimethylsulfoxide (DMSO), or a mixture of 60% M9 and 40% S buffer were used as cryoprotectants. A significantly larger number of juveniles than adults survived deep freezing. Significantly more nematodes were motile after cryopreservation in the 15% glycerol-S-M9 soludon than in the M9-S buffer solution or the DMSO aqueous solution. When cryopreserved nematodes that had been treated with glycerol solution were plated onto Botrytis cinerea, they reproduced rapidly over several generations. Cryopreserved nematodes were as pathogenic as untreated nematodes to Scots pines. PMID:19283151

  9. Inactivation of contaminated fungi and antioxidant effects of peach ( Prunus persica L. Batsch cv Dangeumdo) by 0.5-2 kGy gamma irradiation

    NASA Astrophysics Data System (ADS)

    Kim, Kyoung-Hee; Kim, Mi-Seon; Kim, Hong-Gi; Yook, Hong-Sun

    2010-04-01

    The effect of gamma irradiation (0.5-2 kGy) on the physicochemical properties of peaches was investigated during a 6-day storage at 20±3 °C. Gamma irradiation is able to inactivate the four pathogens, namely Botrytis cinerea, Penicillium expansum, Rhizopus stolonifer var. stolonifer and Monilinia fructicola in peaches. Hardness significantly decreased with the increment of irradiation dose level whereas soluble solid and total polyphenol contents increased with increment of irradiation dose level. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity of the irradiated peach was higher than that of control, and its activity increased with increment of irradiation dose level. These results suggest that gamma irradiation of peaches improved antioxidant activity, but dramatically affects the hardness throughout the entire storage time.

  10. Antifungal Substances from Streptomyces sp. A3265 Antagonistic to Plant Pathogenic Fungi.

    PubMed

    Van Minh, Nguyen; Woo, E-Eum; Kim, Ji-Yul; Kim, Dae-Won; Hwang, Byung Soon; Lee, Yoon-Ju; Lee, In-Kyoung; Yun, Bong-Sik

    2015-09-01

    In a previous study, we identified a Streptomyces sp., A3265, as exhibiting potent antifungal activity against various plant pathogenic fungi, including Botrytis cinerea, Colletotrichum gloeosporioides, and Rhizoctonia solani. This strain also exhibited a biocontrolling effect against ginseng root rot and damping-off disease, common diseases of ginseng and other crops. In this study, we isolated two antifungal substances responsible for this biocontrolling effect via Diaion HP-20 and Sephadex LH-20 column chromatography, medium pressure liquid chromatography, and high-performance liquid chromatography. These compounds were identified as guanidylfungin A and methyl guanidylfungin A by spectroscopic methods. These compounds exhibited potent antimicrobial activity against various plant pathogenic fungi as well as against bacteria. PMID:26539051

  11. A Doped Polyaniline Modified Electrode Amperometric Biosensor for Gluconic Acid Determination in Grapes

    PubMed Central

    Albanese, Donatella; Malvano, Francesca; Sannini, Adriana; Pilloton, Roberto; Di Matteo, Marisa

    2014-01-01

    In winemaking gluconic acid is an important marker for quantitative evaluation of grape infection by Botrytis cinerea. A screen-printed amperometric bienzymatic sensor for the determination of gluconic acid based on gluconate kinase (GK) and 6-phospho-D-gluconate dehydrogenase (6PGDH) coimmobilized onto polyaniline/poly (2-acrylamido-2-methyl-1-propanesulfonic acid; PANI-PAAMPSA) is reported in this study. The conductive polymer electrodeposed on the working electrode surface allowed the detection of NADH at low potential (0.1 V) with a linear range from 4 × 10−3 to 1 mM (R2 = 0.99) and a sensitivity of 419.44 nA·mM−1. The bienzymatic sensor has been optimized with regard to GK/6PGDH enzymatic unit ratio and ATP/NADP+ molar ratio which resulted equal to 0.33 and 1.2, respectively. Under these conditions a sensitivity of 255.2 nA·mM−1, a limit of detection of 5 μM and a Relative Standard Deviation (RSD) of 4.2% (n = 5) have been observed. Finally, the biosensor has been applied for gluconic acid measurements in must grape samples and the matrix effect has been taken into consideration. The results have been compared with those obtained on the same samples with a commercial kit based on a spectrophotometric enzyme assay and were in good agreement, showing the capability of the bienzymatic PANI-PAAMPSA biosensor for gluconic acid measurements and thus for the evaluation of Botrytis cinerea infection in grapes. PMID:24960084

  12. The in-vitro evaluation of antibacterial, antifungal and cytotoxic properties of Marrubium vulgare L. essential oil grown in Tunisia

    PubMed Central

    2011-01-01

    Background In order to validate its antiseptic and anticancer properties with respect to traditional uses, we have screened for the first time the antimicrobial activity of aerial parts of M. vulgare L. essential oil against different pathogenic microorganisms and the cytotoxic activity against HeLa cell lines. Methods The agar disk diffusion method was used to study the antibacterial activity of M. vulgare essential oil against 12 bacterial and 4 fungi strains. The disc diameters of zone of inhibition (DD), the minimum inhibitory concentrations (MIC) and the concentration inhibiting 50% (IC50) were investigated to characterize the antimicrobial activities of this essential oil. The in vitro cytotoxicity of M. vulgare essential oil was examined using a modified MTT assay; the viability and the IC50 were used to evaluate this test. Results The antimicrobial activity of the essential oil was investigated in order to evaluate its efficacy against the different tested microorganisms. The present results results showed a significant activity against microorganisms especially Gram (+) bacteria with inhibition zones and minimal inhibitory concentration values in the range of 6.6-25.2 mm and 1120-2600 μg/ml, respectively, whereas Gram (-) bacteria exhibited a higher resistance. As far as the antifungal activity, among four strains tested, Botrytis cinerea exhibited the strongest activity with inhibition zones of 12.6 mm. However, Fusarium solani, Penicillium digitatum and Aspergillus niger were less sensitive to M. vulgare essential oil. About the citotoxicity assay, this finding indicate the capability of this essential oil to inhibited the proliferation of HeLa cell lines under some conditions with IC50 value of 0.258 μg/ml. Conclusion This investigation showed that the M. vulgare essential oil has a potent antimicrobial activity against some Gram (+) pathogenic bacteria and Botrytis cinerea fungi. The present studies confirm the use of this essential oil as anticancer

  13. Biocontrol Activity of Bacillus amyloliquefaciens CNU114001 against Fungal Plant Diseases.

    PubMed

    Ji, Seung Hyun; Paul, Narayan Chandra; Deng, Jian Xin; Kim, Young Sook; Yun, Bong-Sik; Yu, Seung Hun

    2013-12-01

    A total of 62 bacterial isolates were obtained from Gomsohang mud flat, Mohang mud flat, and Jeju Island, Republic of Korea. Among them, the isolate CNU114001 showed significant antagonistic activity against pathogenic fungi by dual culture method. The isolate CNU114001 was identified as Bacillus amyloliquefaciens by morphological observation and molecular data analysis, including 16SrDNA and gyraseA (gyrA) gene sequences. Antifungal substances of the isolate were extracted and purified by silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. The heat and UV ray stable compound was identified as iturin, a lipopeptide (LP). The isolate CNU114001 showed broad spectrum activity against 12 phytopathogenic fungi by dual culture method. The semi purified compound significantly inhibits the mycelial growth of pathogenic fungi (Alternaria panax, Botrytis cinera, Colletotrichum orbiculare, Penicillium digitatum, Pyricularia grisea and Sclerotinia sclerotiorum) at 200 ppm concentration. Spore germ tube elongation of Botrytis cinerea was inhibited by culture filtrate of the isolate. Crude antifungal substance showed antagonistic activity against cucumber scleotiorum rot in laboratory, and showed antagonistic activity against tomato gray mold, cucumber, and pumpkin powdery mildew in greenhouse condition. PMID:24493945

  14. Circadian clocks and the regulation of virulence in fungi: Getting up to speed.

    PubMed

    Hevia, Montserrat A; Canessa, Paulo; Larrondo, Luis F

    2016-09-01

    You cannot escape time. Therefore, it seems wise to learn how to keep track of it and use it to your advantage. Circadian clocks are molecular circuits that allow organisms to temporally coordinate a plethora of processes, including gene expression, with a close to 24h rhythm, optimizing cellular function in synchrony with daily environmental cycles. The molecular bases of these clocks have been extensively studied in the fungus Neurospora crassa, providing a detailed molecular description. Surprisingly, there is scarce molecular information of clocks in fungi other than Neurospora, despite the existence of rhythmic phenomena in many fungal species, including pathogenic ones. This review will comment on the overall importance of clocks, what is known in Neurospora and what has been described in other fungi including new insights on the evolution of fungal clock components. The molecular description of the circadian system of the phytopathogenic fungus Botrytis cinerea will be revisited, as well as time-of-the-day variation in host-pathogen interaction dynamics, utilizing an Arabidopsis-Botrytis system, including also what is known regarding circadian regulation of defense mechanisms in the Arabidopsis thaliana plant model. Finally, this review will mention how little is known about circadian regulation of human pathogenic fungi, commenting on potential future directions and the overall perspective of fungal circadian studies. PMID:27039027

  15. The effects of some polyamine biosynthetic inhibitors on growth and morphology of phytopathogenic fungi

    NASA Technical Reports Server (NTRS)

    Rajam, M. V.; Galston, A. W.

    1985-01-01

    We have studied the effects of two polyamine biosynthetic inhibitors, alpha-difluoromethylornithine (DFMO) and alpha-difluoromethylarginine (DFMA), and of polyamines (PAs), alone and in combination, on mycelial growth and morphology of four phytopathogenic fungi: Botrytis sp, B. cinerea, Rhizoctonia solani and Monilinia fructicola. The inhibitors were added to a Czapek agar medium to get final concentrations of 0.1, 0.5 and 1.0 mM. DFMO and DFMA, suicide inhibitors of ornithine decarboxylase (ODC) and arginine decarboxylase (ADC) respectively, inhibited mycelial growth strongly; the effect was generally more pronounced with DFMA than with DFMO, but each fungus had its own response pattern. The addition of the PAs putrescine (Put) and spermidine (Spd) to the culture medium resulted in a promotion of growth. In Botrytis sp and Monilinia fructicola exposed to inhibitors plus PAs, mycelial growth was actually increased above control values. Mycelial morphology was altered and cell size dramatically reduced in plates containing inhibitors alone, whereas with PAs alone, or in combination with inhibitors, morphology was normal, but cell length and diameters increased considerably. These results suggest that PAs are essential for growth in fungal mycelia. The inhibition caused by DFMA may be due to its arginase-mediated conversion to DFMO.

  16. Biocontrol Activity of Bacillus amyloliquefaciens CNU114001 against Fungal Plant Diseases

    PubMed Central

    Ji, Seung Hyun; Paul, Narayan Chandra; Deng, Jian Xin; Kim, Young Sook; Yun, Bong-Sik

    2013-01-01

    A total of 62 bacterial isolates were obtained from Gomsohang mud flat, Mohang mud flat, and Jeju Island, Republic of Korea. Among them, the isolate CNU114001 showed significant antagonistic activity against pathogenic fungi by dual culture method. The isolate CNU114001 was identified as Bacillus amyloliquefaciens by morphological observation and molecular data analysis, including 16SrDNA and gyraseA (gyrA) gene sequences. Antifungal substances of the isolate were extracted and purified by silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. The heat and UV ray stable compound was identified as iturin, a lipopeptide (LP). The isolate CNU114001 showed broad spectrum activity against 12 phytopathogenic fungi by dual culture method. The semi purified compound significantly inhibits the mycelial growth of pathogenic fungi (Alternaria panax, Botrytis cinera, Colletotrichum orbiculare, Penicillium digitatum, Pyricularia grisea and Sclerotinia sclerotiorum) at 200 ppm concentration. Spore germ tube elongation of Botrytis cinerea was inhibited by culture filtrate of the isolate. Crude antifungal substance showed antagonistic activity against cucumber scleotiorum rot in laboratory, and showed antagonistic activity against tomato gray mold, cucumber, and pumpkin powdery mildew in greenhouse condition. PMID:24493945

  17. Genetically engineered Thompson Seedless grapevine plants designed for fungal tolerance: selection and characterization of the best performing individuals in a field trial.

    PubMed

    Rubio, Julia; Montes, Christian; Castro, Álvaro; Álvarez, Catalina; Olmedo, Blanca; Muñoz, Marisol; Tapia, Eduardo; Reyes, Fernando; Ortega, Marcelo; Sánchez, Evelyn; Miccono, María; Dalla Costa, Lorenza; Martinelli, Lucia; Malnoy, Mickael; Prieto, Humberto

    2015-02-01

    The fungi Botrytis cinerea and Erysiphe necator are responsible for gray mold and powdery mildew diseases, respectively, which are among the most devastating diseases of grapes. Two endochitinase (ech42 and ech33) genes and one N-acetyl-β-D-hexosaminidase (nag70) gene from biocontrol agents related to Trichoderma spp. were used to develop a set of 103 genetically modified (GM) 'Thompson Seedless' lines (568 plants) that were established in open field in 2004 and evaluated for fungal tolerance starting in 2006. Statistical analyses were carried out considering transgene, explant origin, and plant response to both fungi in the field and in detached leaf assays. The results allowed for the selection of the 19 consistently most tolerant lines through two consecutive years (2007-2008 and 2008-2009 seasons). Plants from these lines were grafted onto the rootstock Harmony and established in the field in 2009 for further characterization. Transgene status was shown in most of these lines by Southern blot, real-time PCR, ELISA, and immunostrips; the most tolerant candidates expressed the ech42-nag70 double gene construct and the ech33 gene from a local Hypocrea virens isolate. B. cinerea growth assays in Petri dishes supplemented with berry juices extracted from the most tolerant individuals of the selected population was inhibited. These results demonstrate that improved fungal tolerance can be attributed to transgene expression and support the iterative molecular and physiological phenotyping in order to define selected individuals from a population of GM grapevines. PMID:25011563

  18. Effect of Selenium on Control of Postharvest Gray Mold of Tomato Fruit and the Possible Mechanisms Involved.

    PubMed

    Wu, Zhilin; Yin, Xuebin; Bañuelos, Gary S; Lin, Zhi-Qing; Zhu, Zhu; Liu, Ying; Yuan, Linxi; Li, Miao

    2015-01-01

    Selenium (Se) has important benefits for crop growth and stress tolerance at low concentrations. However, there is very little information on antimicrobial effect of Se against the economically important fungus Botrytis cinerea. In the present study, using sodium selenite as Se source, we investigated the effect of Se salts on spore germination and mycelial growth of the fungal pathogen in vitro and gray mold control in harvested tomato fruit. Se treatment at 24 mg/L significantly inhibited spore germination of the fungal pathogen and effectively controlled gray mold in harvested tomato fruit. Se treatment at 24 mg/L seems to induce the generation of intracellular reactive oxygen species in the fungal spores. The membrane integrity damage was observed with fluorescence microscopy following staining with propidium iodide after treatment of the spores with Se. These results suggest that Se has the potential for controlling gray mold rot of tomato fruits and might be useful in integrated control against gray mold disease of postharvest fruits and vegetables caused by B. cinerea. The mechanisms by which Se decreased gray mold decay of tomato fruit may be directly related to the severe damage to the conidia plasma membrane and loss of cytoplasmic materials from the hyphae. PMID:26779128

  19. In Vitro Control of Post-Harvest Fruit Rot Fungi by Some Plant Essential Oil Components