Occurrence of Leaf Blight on Cosmos Caused by Alternaria cosmosa in Korea.

PubMed

Deng, Jian Xin; Lee, Ji Hye; Paul, Narayan Chandra; Cho, Hye Sun; Lee, Hyang Burm; Yu, Seung Hun

2015-03-01

In 2011, a leaf blight disease was observed on cosmos (Cosmos bipinnatus) leaves in Nonsan, Korea. The causal pathogen was isolated and identified based on morphological and molecular approaches. Morphological characteristics of the pathogen matched well with the Alternaria cosmosa and also easily distinguishable from Alternaria zinniae reported from cosmos seeds by producing branched beak. Phylogenetically, the pathogen could not be distinguished from A. passiflorae based on the sequence analysis of a combined data set of Alt a1 and gpd genes. However, A. passiflorae was distinguished from the present species by having conidiophores with 4 to 5 conidiogenous loci. The results indicate that the present Alternaria species is A. cosmosa. Pathogenicity tests revealed that the isolate was pathogenic to the leaves of Cosmos bipinnatus. This is the first report of Alternaria blight disease caused by A. cosmosa on cosmos in Korea.

Occurrence of Leaf Blight on Cosmos Caused by Alternaria cosmosa in Korea

PubMed Central

Deng, Jian Xin; Lee, Ji Hye; Paul, Narayan Chandra; Cho, Hye Sun; Lee, Hyang Burm; Yu, Seung Hun

2015-01-01

In 2011, a leaf blight disease was observed on cosmos (Cosmos bipinnatus) leaves in Nonsan, Korea. The causal pathogen was isolated and identified based on morphological and molecular approaches. Morphological characteristics of the pathogen matched well with the Alternaria cosmosa and also easily distinguishable from Alternaria zinniae reported from cosmos seeds by producing branched beak. Phylogenetically, the pathogen could not be distinguished from A. passiflorae based on the sequence analysis of a combined data set of Alt a1 and gpd genes. However, A. passiflorae was distinguished from the present species by having conidiophores with 4 to 5 conidiogenous loci. The results indicate that the present Alternaria species is A. cosmosa. Pathogenicity tests revealed that the isolate was pathogenic to the leaves of Cosmos bipinnatus. This is the first report of Alternaria blight disease caused by A. cosmosa on cosmos in Korea. PMID:25774114

Nuclear Ribosomal DNA Variation and Pathogenic Specialization in Alternaria Fungi Known To Produce Host-Specific Toxins †

PubMed Central

Kusaba, Motoaki; Tsuge, Takashi

1994-01-01

A total of 99 strains of 11 Alternaria species, including 68 strains of seven fungi known to produce host-specific toxins, were subjected to analysis of restriction fragment length polymorphism (RFLP) in nuclear ribosomal DNA (rDNA). Total DNA was digested with XbaI, and the Southern blots were probed with a nuclear rDNA clone of Alternaria kikuchiana. The hybridization gave 17 different RFLPs from the 99 strains. On the basis of these RFLPs, populations of host-specific toxin-producing fungi could not be differentiated from one another nor from nonpathogenic A. alternata. Each population of the toxin-producing fungi carried rDNA variants. Nine different types, named A1 to A6 and B1 to B3, were detected among the toxin-producing fungi and nonpathogenic A. alternata. All of the populations contained the type A4 variant, and the other rDNA types were also shared by different toxin-producing fungi and A. alternata. In contrast, Alternaria species that are morphologically distinguishable from A. alternata could be differentiated from A. alternata on the basis of the rDNA RFLPs. Polymorphisms in rDNA digested with HaeIII and MspI were also evaluated in 61 Alternaria strains. These restriction enzymes produced 31 variations among all of the samples. The seven toxin-producing fungi and nonpathogenic A. alternata could not be resolved by phylogenetic analysis based on the RFLPs, although they could be differentiated from the other Alternaria species studied. These results provide support for the hypothesis that Alternaria fungi known to produce host-specific toxins are intraspecific variants of A. alternata specialized in pathogenicity. Images PMID:16349367

Comparison of pathogenicity of Alternaria pellucida and Curvularia lunata on weed Echinochloa species.

PubMed

Reza, Mohammad; Motlagh, Safari

2015-07-01

Echinochloa spp. are the most important weeds in rice fields. In this research Curvularia lunata and Alternaria pellucida were isolated from these weeds and their pathogenicity effects were compared on these weeds and five rice cultivars in a completely random design with three replications in greenhouse conditions. Fungi were inoculated on weeds and rice cultivars, using spore suspension consisting of 10' spore ml(-1) of distilled water. Results indicated significant effect of Curvularia lunata and Alternaria pellucida on Echinochloa oryzicola and E. crus-galli. In the present study, effect of C. lunata on fresh weight, dry weight and height of Echinochloa species based on variance analysis table, a significant reaction was observed for height and fresh weight, but for dry weight reaction was not significant. The effect of A. pellucida on fresh weight, dry weight and height of Echinochloa species based on variance analysis table, a significant reaction was observed for all the three traits. Also, rice cultivars did not show any significant reaction to C. lunata and A. pellucida. The results showed that in comparison between effect of Curvularia lunata and Alternaria pellucida on Echinochloa spp., disease rating caused by A. pellucida on E. oryzicola and E. crusalli was more than disease rating caused by C. lunata and these species of weed were more susceptible to A. pellucida. However, A. alternata can be considered as a better promising bioherbicide to control Echinochloa spp.

Protein interaction networks at the host-microbe interface in Diaphorina citri, the insect vector of the citrus greening pathogen

USDA-ARS?s Scientific Manuscript database

The Asian citrus psyllid (Diaphorina citri) is the insect vector responsible for the worldwide spread of Candidatus Liberibacter asiaticus, the bacterial pathogen associated with citrus greening disease. Developmental changes in the insect vector impact pathogen transmission, such that D. citri tra...

Fungicidal activity of silver nanoparticles against Alternaria brassicicola

NASA Astrophysics Data System (ADS)

Gupta, Deepika; Chauhan, Pratima

2016-04-01

This work highlighted the fungicidal properties of silver nanoparticles against Alternaria brassicicola. Alternaria brassicicola causes Black spot of Cauliflower, radish, cabbage, kale which results in sever agricultural loss. We treat the synthesised silver nanoparticles (AgNPs) of 10, 25, 50, 100 and 110 ppm concentrations against Alternaria brassicicola on PDA containing Petri dish. We calculated inhibitory rate (%) in order to evaluate the antifungal efficacy of silver nanoparticles against pathogens. Treatment with 100ppm AgNPs resulted in maximum inhibition of Alternaria brassicicola i.e.92.2%. 110ppm of AgNPS also shows the same result, therefore 100ppm AgNPs was treated as optimize concentration. AgNPs effectively inhibited the growth of a Alternaria brassicicola, which suggests that AgNPs could be used as fungicide in plant disease management. Further research and development are necessary to translate this technology into plant disease management strategies.

Alternaria in Food: Ecophysiology, Mycotoxin Production and Toxicology

PubMed Central

Patriarca, Andrea; Magan, Naresh

2015-01-01

Alternaria species are common saprophytes or pathogens of a wide range of plants pre- and post-harvest. This review considers the relative importance of Alternaria species, their ecology, competitiveness, production of mycotoxins and the prevalence of the predominant mycotoxins in different food products. The available toxicity data on these toxins and the potential future impacts of Alternaria species and their toxicity in food products pre- and post-harvest are discussed. The growth of Alternaria species is influenced by interacting abiotic factors, especially water activity (aw), temperature and pH. The boundary conditions which allow growth and toxin production have been identified in relation to different matrices including cereal grain, sorghum, cottonseed, tomato, and soya beans. The competitiveness of Alternaria species is related to their water stress tolerance, hydrolytic enzyme production and ability to produce mycotoxins. The relationship between A. tenuissima and other phyllosphere fungi has been examined and the relative competitiveness determined using both an Index of Dominance (ID) and the Niche Overlap Index (NOI) based on carbon-utilisation patterns. The toxicology of some of the Alternaria mycotoxins have been studied; however, some data are still lacking. The isolation of Alternaria toxins in different food products including processed products is reviewed. The future implications of Alternaria colonization/infection and the role of their mycotoxins in food production chains pre- and post-harvest are discussed. PMID:26190916

Structural and biochemical characteristics of citrus flowers associated with defence against a fungal pathogen

PubMed Central

Marques, João Paulo Rodrigues; Amorim, Lilian; Silva-Junior, Geraldo José; Spósito, Marcel Bellato; Appezzato-da Gloria, Beatriz

2015-01-01

The constitutive characters of plants can be structural or biochemical and play an important role in their defence against pathogens. Citrus postbloom fruit drop (PFD) caused by Colletotrichum spp. is one of the most important fungal diseases of citrus. The pathogen infects the flowers, leading to premature fruit drop and reducing citrus production. However, flower buds smaller than 8 mm long are usually not infected by Colletotrichum spp. Thus, this study investigated whether there are constitutive mechanisms in flower buds related to Colletotrichum spp. infection. We studied flower buds that were 2, 3, 4, 8, 12 and 15 mm long and petals, after anthesis, of sweet orange ‘Valência’ using light and scanning electron microscopy and histochemistry. We evaluated the effect of volatile organic compounds (VOCs) in flowers (R-limonene and linalool) on the in vitro growth of Colletotrichum acutatum. We found that the arrangement of the epidermal papillae in the petal primordia, the occurrence of prismatic crystals and the distribution of oil glands are the main differences between buds smaller than 8 mm and buds 8–15 mm long. Osmophores at the tips of petals produced and accumulated phenols, terpenes and lipophilic compounds. Flower buds smaller than 8 mm long have constitutive structural and biochemical barriers to Colletotrichum spp. infection. In addition, this is the first time that osmophores have been reported in citrus. Our study shows that natural terpenes of Citrus flowers inhibit the fungal growth in vitro, highlighting the potential use of terpenes for the chemical control of PFD in citrus. PMID:25535209

A Pathogen Secreted Protein as a Detection Marker for Citrus Huanglongbing

PubMed Central

Pagliaccia, Deborah; Shi, Jinxia; Pang, Zhiqian; Hawara, Eva; Clark, Kelley; Thapa, Shree P.; De Francesco, Agustina D.; Liu, Jianfeng; Tran, Thien-Toan; Bodaghi, Sohrab; Folimonova, Svetlana Y.; Ancona, Veronica; Mulchandani, Ashok; Coaker, Gitta; Wang, Nian; Vidalakis, Georgios; Ma, Wenbo

2017-01-01

The citrus industry is facing an unprecedented crisis due to Huanglongbing (HLB, aka citrus greening disease), a bacterial disease associated with the pathogen Candidatus Liberibacter asiaticus (CLas) that affects all commercial varieties. Transmitted by the Asian citrus psyllid (ACP), CLas colonizes citrus phloem, leading to reduced yield and fruit quality, and eventually tree decline and death. Since adequate curative measures are not available, a key step in HLB management is to restrict the spread of the disease by identifying infected trees and removing them in a timely manner. However, uneven distribution of CLas cells in infected trees and the long latency for disease symptom development makes sampling of trees for CLas detection challenging. Here, we report that a CLas secreted protein can be used as a biomarker for detecting HLB infected citrus. Proteins secreted from CLas cells can presumably move along the phloem, beyond the site of ACP inoculation and CLas colonized plant cells, thereby increasing the chance of detecting infected trees. We generated a polyclonal antibody that effectively binds to the secreted protein and developed serological assays that can successfully detect CLas infection. This work demonstrates that antibody-based diagnosis using a CLas secreted protein as the detection marker for infected trees offers a high-throughput and economic approach that complements the approved quantitative polymerase chain reaction-based methods to enhance HLB management programs. PMID:29403441

Furfural from Pine Needle Extract Inhibits the Growth of a Plant Pathogenic Fungus, Alternaria mali

PubMed Central

Yoo, Sun Kyun; Moon, Sung-Kwon; Lee, Ung-Soo

2007-01-01

The antifungal effect of pine needle extract prepared by a distinguishable extraction method and the dry distillation method, was examined. The effect of this extract itself was insignificant. The chemical components of pine needle extract were then investigated by gas chromatographic analysis, and four chemical components, acetol, furfural, 5-methyl furfural, and terpine-4-ol, were identified. The antifungal effects of those four chemical components against Alternaria mali (A. mali), an agent of Alternaria blotch of apple, were then examined. It was observed that the minimum inhibitory concentrations (MICs) were 6.25, 0.78, 0.78, and 12.5 (mg/ml) of acetol, furfural, 5-methyl furfural, and terpine-4-ol, respectively. MICs of furfural and 5-methyl furfural had the same order of magnitude as that of an antifungal agrochemical, chlorothalonil. Although furfural itself can not be completely substituted for an antifungal agrochemical, a partial mixture of furfural and antifungal agrochemical may be used as a substitute. The use of agrochemicals for the prevention of plant disease caused by pathogenic fungus such as A. mali could be partially reduced by the application of this mixture. PMID:24015067

The destructive citrus pathogen, ‘Candidatus Liberibacter asiaticus’ encodes a functional flagellin characteristic of a pathogen-associated molecular pattern

USDA-ARS?s Scientific Manuscript database

Huanglongbing (HLB) is presently the most devastating citrus disease worldwide. As an intracellular plant pathogen and insect symbiont, the HLB bacterium, ‘Candidatus Liberibacter asiaticus’ (Las) retains the entire flagellum-encoding gene cluster in its significantly reduced genome. Las encodes a...

Mould and yeast flora in fresh berries, grapes and citrus fruits.

PubMed

Tournas, V H; Katsoudas, Eugenia

2005-11-15

Fresh fruits are prone to fungal contamination in the field, during harvest, transport, marketing, and with the consumer. It is important to identify fungal contaminants in fresh fruits because some moulds can grow and produce mycotoxins on these commodities while certain yeasts and moulds can cause infections or allergies. In this study, 251 fresh fruit samples including several varieties of grapes, strawberries, blueberries, raspberries, blackberries, and various citrus fruits were surface-disinfected, incubated at room temperature for up to 14 days without supplemental media, and subsequently examined for mould and yeast growth. The level of contamination (percent of contaminated items/sample) varied depending on the type of fruit. All raspberry and blackberry samples were contaminated at levels ranging from 33% to 100%, whereas 95% of the blueberry samples supported mould growth at levels between 10% and 100% of the tested berries, and 97% of strawberry samples showed fungal growth on 33-100% of tested berries. The most common moulds isolated from these commodities were Botrytis cinerea, Rhizopus (in strawberries), Alternaria, Penicillium, Cladosporium and Fusarium followed by yeasts, Trichoderma and Aureobasidium. Thirty-five percent of the grape samples tested were contaminated and supported fungal growth; the levels of contamination ranged from 9% to 80%. The most common fungi spoiling grapes were Alternaria, B. cinerea and Cladosporium. Eighty-three percent of the citrus fruit samples showed fungal growth at levels ranging from 25% to 100% of tested fruits. The most common fungi in citrus fruits were Alternaria, Cladosporium, Penicillium, Fusarium and yeasts. Less common were Trichoderma, Geotrichum and Rhizopus.

Genomic and transcriptomic analyses of the tangerine pathotype of Alternaria alternata in response to oxidative stress

PubMed Central

Wang, Mingshuang; Sun, Xuepeng; Yu, Dongliang; Xu, Jianping; Chung, Kuangren; Li, Hongye

2016-01-01

The tangerine pathotype of Alternaria alternata produces the A. citri toxin (ACT) and is the causal agent of citrus brown spot that results in significant yield losses worldwide. Both the production of ACT and the ability to detoxify reactive oxygen species (ROS) are required for A. alternata pathogenicity in citrus. In this study, we report the 34.41 Mb genome sequence of strain Z7 of the tangerine pathotype of A. alternata. The host selective ACT gene cluster in strain Z7 was identified, which included 25 genes with 19 of them not reported previously. Of these, 10 genes were present only in the tangerine pathotype, representing the most likely candidate genes for this pathotype specialization. A transcriptome analysis of the global effects of H2O2 on gene expression revealed 1108 up-regulated and 498 down-regulated genes. Expressions of those genes encoding catalase, peroxiredoxin, thioredoxin and glutathione were highly induced. Genes encoding several protein families including kinases, transcription factors, transporters, cytochrome P450, ubiquitin and heat shock proteins were found associated with adaptation to oxidative stress. Our data not only revealed the molecular basis of ACT biosynthesis but also provided new insights into the potential pathways that the phytopathogen A. alternata copes with oxidative stress. PMID:27582273

Infection Density Dynamics of the Citrus Greening Bacterium “Candidatus Liberibacter asiaticus” in Field Populations of the Psyllid Diaphorina citri and Its Relevance to the Efficiency of Pathogen Transmission to Citrus Plants

PubMed Central

Ukuda-Hosokawa, Rie; Sadoyama, Yasutsune; Kishaba, Misaki; Kuriwada, Takashi; Anbutsu, Hisashi

2015-01-01

Huanglongbing, or citrus greening, is a devastating disease of citrus plants recently spreading worldwide, which is caused by an uncultivable bacterial pathogen, “Candidatus Liberibacter asiaticus,” and vectored by a phloem-sucking insect, Diaphorina citri. We investigated the infection density dynamics of “Ca. Liberibacter asiaticus” in field populations of D. citri with experiments using field-collected insects to address how “Ca. Liberibacter asiaticus” infection density in the vector insect is relevant to pathogen transmission to citrus plants. Of 500 insects continuously collected from “Ca. Liberibacter asiaticus”-infected citrus trees with pathological symptoms in the spring and autumn of 2009, 497 (99.4%) were “Ca. Liberibacter asiaticus” positive. The infections were systemic across head-thorax and abdomen, ranging from 103 to 107 bacteria per insect. In spring, the infection densities were low in March, at ∼103 bacteria per insect, increasing up to 106 to 107 bacteria per insect in April and May, and decreasing to 105 to 106 bacteria per insect in late May, whereas the infection densities were constantly ∼106 to 107 bacteria per insect in autumn. Statistical analysis suggested that several factors, such as insect sex, host trees, and collection dates, may be correlated with “Ca. Liberibacter asiaticus” infection densities in field D. citri populations. Inoculation experiments with citrus seedlings using field-collected “Ca. Liberibacter asiaticus”-infected insects suggested that (i) “Ca. Liberibacter asiaticus”-transmitting insects tend to exhibit higher infection densities than do nontransmitting insects, (ii) a threshold level (∼106 bacteria per insect) of “Ca. Liberibacter asiaticus” density in D. citri is required for successful transmission to citrus plants, and (iii) D. citri attaining the threshold infection level transmits “Ca. Liberibacter asiaticus” to citrus plants in a stochastic manner. These

Infection Density Dynamics of the Citrus Greening Bacterium "Candidatus Liberibacter asiaticus" in Field Populations of the Psyllid Diaphorina citri and Its Relevance to the Efficiency of Pathogen Transmission to Citrus Plants.

PubMed

Ukuda-Hosokawa, Rie; Sadoyama, Yasutsune; Kishaba, Misaki; Kuriwada, Takashi; Anbutsu, Hisashi; Fukatsu, Takema

2015-06-01

Huanglongbing, or citrus greening, is a devastating disease of citrus plants recently spreading worldwide, which is caused by an uncultivable bacterial pathogen, "Candidatus Liberibacter asiaticus," and vectored by a phloem-sucking insect, Diaphorina citri. We investigated the infection density dynamics of "Ca. Liberibacter asiaticus" in field populations of D. citri with experiments using field-collected insects to address how "Ca. Liberibacter asiaticus" infection density in the vector insect is relevant to pathogen transmission to citrus plants. Of 500 insects continuously collected from "Ca. Liberibacter asiaticus"-infected citrus trees with pathological symptoms in the spring and autumn of 2009, 497 (99.4%) were "Ca. Liberibacter asiaticus" positive. The infections were systemic across head-thorax and abdomen, ranging from 10(3) to 10(7) bacteria per insect. In spring, the infection densities were low in March, at ∼ 10(3) bacteria per insect, increasing up to 10(6) to 10(7) bacteria per insect in April and May, and decreasing to 10(5) to 10(6) bacteria per insect in late May, whereas the infection densities were constantly ∼ 10(6) to 10(7) bacteria per insect in autumn. Statistical analysis suggested that several factors, such as insect sex, host trees, and collection dates, may be correlated with "Ca. Liberibacter asiaticus" infection densities in field D. citri populations. Inoculation experiments with citrus seedlings using field-collected "Ca. Liberibacter asiaticus"-infected insects suggested that (i) "Ca. Liberibacter asiaticus"-transmitting insects tend to exhibit higher infection densities than do nontransmitting insects, (ii) a threshold level (∼ 10(6) bacteria per insect) of "Ca. Liberibacter asiaticus" density in D. citri is required for successful transmission to citrus plants, and (iii) D. citri attaining the threshold infection level transmits "Ca. Liberibacter asiaticus" to citrus plants in a stochastic manner. These findings provide

Induced resistance in tomato fruit by γ-aminobutyric acid for the control of alternaria rot caused by Alternaria alternata.

PubMed

Yang, Jiali; Sun, Cui; Zhang, Yangyang; Fu, Da; Zheng, Xiaodong; Yu, Ting

2017-04-15

The study investigated the effect of γ-aminobutyric acid (GABA) on the control of alternaria rot in tomato fruit and the possible mechanism involved. Our results showed exogenous GABA could stimulate remarkable resistance to the alternaria rot, while it had no direct antifungal activity against Alternaria alternata. Moreover, the activities of antioxidant enzymes, including peroxidase, superoxide dismutase and catalase, along with the expression of these corresponding genes, were significantly induced in the GABA treatment. The obtained data suggested GABA induced resistance against the necrotrophic pathogen A. alternata, at least in part by activating antioxidant enzymes, restricting the levels of cell death caused by reactive oxygen species. Meanwhile, the key enzyme genes of GABA shunt, GABA transaminase and succinic-semialdehyde dehydrogenase, were found up-regulated in the GABA treatment. The activation of the GABA shunt might play a vital role in the resistance mechanism underpinning GABA-induced plant immunity. Copyright © 2016 Elsevier Ltd. All rights reserved.

Fungal-specific transcription factor AbPf2 activates pathogenicity in Alternaria brassicicola

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cho, Yangrae; Ohm, Robin A.; Grigoriev, Igor V.

Alternaria brassicicola is a successful saprophyte and necrotrophic plant pathogen. To identify molecular determinants of pathogenicity, we created non-pathogenic mutants of a transcription factor-encoding gene, AbPf2. The frequency and timing of germination and appressorium formation on host plants were similar between the non-pathogenic abpf2 mutants and wild-type A. brassicicola. The mutants were also similar in vitro to wild-type A. brassicicola in terms of vegetative growth, conidium production, and responses to a phytoalexin, reactive oxygen species and osmolites. The hyphae of the mutants grew slowly but did not cause disease symptoms on the surface of host plants. Transcripts of the AbPf2more » gene increased exponentially soon after wild-type conidia contacted their host plants . A small amount of AbPf2 protein, as monitored using GFP fusions, was present in young, mature conidia. The protein level decreased during saprophytic growth, but increased and was located primarily in fungal nuclei during pathogenesis. Levels of the proteins and transcripts sharply decreased following colonization of host tissues beyond the initial infection site. When expression of the transcription factor was induced in the wild-type during early pathogenesis, 106 fungal genes were also induced in the wild-type but not in the abpf2 mutants. Notably, 33 of the 106 genes encoded secreted proteins, including eight putative effector proteins. Plants inoculated with abpf2 mutants expressed higher levels of genes associated with photosynthesis, the pentose phosphate pathway and primary metabolism, but lower levels of defense-related genes. Our results suggest that AbPf2 is an important regulator of pathogenesis, but does not affect other cellular processes in A. brassicicola.« less

Bacterial diversity analysis of Huanglongbing pathogen-infected citrus, using PhyloChip and 16S rRNA gene clone library sequencing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shankar Sagaram, U.; DeAngelis, K.M.; Trivedi, P.

The bacterial diversity associated with citrus leaf midribs was characterized 1 from citrus groves that contained the Huanglongbing (HLB) pathogen, which has yet to be cultivated in vitro. We employed a combination of high-density phylogenetic 16S rDNA microarray and 16S rDNA clone library sequencing to determine the microbial community composition of symptomatic and asymptomatic citrus midribs. Our results revealed that citrus leaf midribs can support a diversity of microbes. PhyloChip analysis indicated that 47 orders of bacteria from 15 phyla were present in the citrus leaf midribs while 20 orders from phyla were observed with the cloning and sequencing method.more » PhyloChip arrays indicated that nine taxa were significantly more abundant in symptomatic midribs compared to asymptomatic midribs. Candidatus Liberibacter asiaticus (Las) was detected at a very low level in asymptomatic plants, but was over 200 times more abundant in symptomatic plants. The PhyloChip analysis was further verified by sequencing 16S rDNA clone libraries, which indicated the dominance of Las in symptomatic leaves. These data implicate Las as the pathogen responsible for HLB disease. Citrus is the most important commercial fruit crop in Florida. In recent years, citrus Huanglongbing (HLB), also called citrus greening, has severely affected Florida's citrus production and hence has drawn an enormous amount of attention. HLB is one of the most devastating diseases of citrus (6,13), characterized by blotchy mottling with green islands on leaves, as well as stunting, fruit decline, and small, lopsided fruits with poor coloration. The disease tends to be associated with a phloem-limited fastidious {alpha}-proteobacterium given a provisional Candidatus status (Candidatus Liberobacter spp. later changed to Candidatus Liberibacter spp.) in nomenclature (18,25,34). Previous studies indicate that HLB infection causes disorder in the phloem and severely impairs the translocation of assimilates

Diaphorina citri (Hemiptera: Liviidae) Vector Competence for the Citrus Greening Pathogen 'Candidatus Liberibacter Asiaticus'.

PubMed

Tabachnick, Walter J

2015-06-01

Characterizing the vector competence of Diaphorina citri Kuwayama for 'Candidatus Liberibacter asiaticus,' the pathogen causing citrus greening, is essential for understanding the epidemiology of this disease that is threatening the U.S. citrus industry. Vector competence studies have been difficult because of the biology of D. citri, the inability to culture the pathogen, and the available diagnostic methods used to detect the bacteria in plant and insect tissues. The methods employed in many studies of D. citri vector competence may have overestimated amounts of live 'Ca. L. asiaticus' in both plant and insect tissues, and it is possible that the amounts of phloem ingested by psyllids may not contain sufficient detectable pathogen using current diagnostic methods. As a result of the difficulty in characterizing D. citri vector competence, the several daunting challenges for providing D. citri that are unable to inoculate 'Ca. L. asiaticus', as a novel method to control greening are discussed. Suggestions to overcome some of these challenges are provided. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Secreted protein extract analyses present the plant pathogen Alternaria alternata as a suitable industrial enzyme toolbox.

PubMed

García-Calvo, L; Ullán, R V; Fernández-Aguado, M; García-Lino, A M; Balaña-Fouce, R; Barreiro, C

2018-04-15

byproducts and analogous substrates. Moreover, the proteomic analysis of the secretome of a natural isolate of Alternaria sp. grown in the presence of one of the most used vegetal substrates on the biofuels industry (sugar beet pulp) sheds light on the extracellular enzymatic machinery of this fungal plant pathogen, and can be potentially applied to developing new industrial enzymatic tools. This work is, to our knowledge, the first to analyze in depth the secreted enzyme extract of the plant pathogen Alternaria when grown on a lignocellulosic substrate, identifying its proteins by means of MALDI-TOF/TOF mass spectrometry and characterizing its feruloyl esterase, cellulase and xylanolytic activities. Copyright © 2018 Elsevier B.V. All rights reserved.

Regulation of pathogenicity in hop stunt viroid-related group II citrus viroids.

PubMed

Reanwarakorn, K; Semancik, J S

1998-12-01

Nucleotide sequences were determined for two hop stunt viroid-related Group II citrus viroids characterized as either a cachexia disease non-pathogenic variant (CVd-IIa) or a pathogenic variant (CVd-IIb). Sequence identity between the two variants of 95.6% indicated a conserved genome with the principal region of nucleotide difference clustered in the variable (V) domain. Full-length viroid RT-PCR cDNA products were cloned into plasmid SP72. Viroid cDNA clones as well as derived RNA transcripts were transmissible to citron (Citrus medica L.) and Luffa aegyptiaca Mill. To determine the locus of cachexia pathogenicity as well as symptom expression in Luffa, chimeric viroid cDNA clones were constructed from segments of either the left terminal, pathogenic and conserved (T1-P-C) domains or the conserved, variable and right terminal (C-V-T2) domains of CVd-IIa or CVd-IIb in reciprocal exchanges. Symptoms induced by the various chimeric constructs on the two bioassay hosts reflected the differential response observed with CVd-IIa and -IIb. Constructs with the C-V-T2 domains region from clone-IIa induced severe symptoms on Luffa typical of CVd-IIa, but were non-symptomatic on mandarin as a bioassay host for the cachexia disease. Constructs with the same region (C-V-T2) from the clone-IIb genome induced only mild symptoms on Luffa, but produced a severe reaction on mandarin, as observed for CVd-IIb. Specific site-directed mutations were introduced into the V domain of the CVd-IIa clone to construct viroid cDNA clones with either partial or complete conversions to the CVd-IIb sequence. With the introduction of six site-specific changes into the V domain of the clone-IIa genome, cachexia pathogenicity was acquired as well as a moderation of severe symptoms on Luffa.

Antennal and behavioral response of the Asian citrus psyllid (Diaphorina citri Kuwayama) to degradation products of citrus volatiles

USDA-ARS?s Scientific Manuscript database

Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae) vectors the bacterial causal pathogen of the deadly citrus disease, Huanglongbing (citrus greening) which is a major threat to citrus industry worldwide. We studied antennal and behavioral responses to principal components of head...

Pseudomonas caspiana sp. nov., a citrus pathogen in the Pseudomonas syringae phylogenetic group.

PubMed

Busquets, Antonio; Gomila, Margarita; Beiki, Farid; Mulet, Magdalena; Rahimian, Heshmat; García-Valdés, Elena; Lalucat, Jorge

2017-07-01

In a screening by multilocus sequence analysis of Pseudomonas strains isolated from diverse origins, 4 phylogenetically closely related strains (FBF58, FBF102 T , FBF103, and FBF122) formed a well-defined cluster in the Pseudomonas syringae phylogenetic group. The strains were isolated from citrus orchards in northern Iran with disease symptoms in the leaves and stems and its pathogenicity against citrus plants was demonstrated. The whole genome of the type strain of the proposed new species (FBF102 T =CECT 9164 T =CCUG 69273 T ) was sequenced and characterized. Comparative genomics with the 14 known Pseudomonas species type strains of the P. syringae phylogenetic group demonstrated that this strain belonged to a new genomic species, different from the species described thus far. Genome analysis detected genes predicted to be involved in pathogenesis, such as an atypical type 3 secretion system and two type 6 secretion systems, together with effectors and virulence factors. A polyphasic taxonomic characterization demonstrated that the 4 plant pathogenic strains represented a new species, for which the name Pseudomonas caspiana sp. nov. is proposed. Copyright © 2017 Elsevier GmbH. All rights reserved.

Huanglongbing increases Diplodia Stem End Rot in Citrus sinensis

USDA-ARS?s Scientific Manuscript database

Huanglongbing (HLB), one of the most devastating diseases of citrus is caused by the a-Proteobacteria Candidatus Liberibacter. Diplodia natalensis Pole-Evans is a fungal pathogen which has been known to cause a postharvest stem-end rot of citrus, the pathogen infects citrus fruit under the calyx, an...

Overexpression of a modified plant thionin enhances disease resistance to citrus canker and huanglongbing (HLB, citrus greening)

USDA-ARS?s Scientific Manuscript database

Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the United States citrus industry. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri). In this study, we characterized e...

Effect of citrus pulp on the viability of Saccharomyces boulardii in the presence of enteric pathogens

USDA-ARS?s Scientific Manuscript database

Saccharomyces cerevisiae subtype boulardii is frequently used as a dietary supplement to promote intestinal health and reduce the impact of growth of enteric pathogens in livestock, including cattle and swine. Citrus by-products are also fed as dietary supplements that have the additional benefit o...

Fungicide sensitivity of Alternaria spp. causing Alternaria leaf spot on sugarbeet in Michigan

USDA-ARS?s Scientific Manuscript database

Alternaria leaf spot (ALS), caused by Alternaria spp., can occur wherever sugarbeet is grown. Infection by Alternaria spp. and disease management has historically been considered a minor issue in sugarbeet production in the US. Recently, increased incidence and severity of ALS has been observed in M...

Phagostimulants for the Asian citrus psyllid also elicit volatile release from citrus leaves

USDA-ARS?s Scientific Manuscript database

Chemical cues that elicit orientation by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), are of great interest because it is the primary vector of the causal pathogen of citrus greening disease. We identified an optimal blend ratio of formic and acetic acids that stimulate...

Characterization of glutathione transferases involved in the pathogenicity of Alternaria brassicicola.

PubMed

Calmes, Benoit; Morel-Rouhier, Mélanie; Bataillé-Simoneau, Nelly; Gelhaye, Eric; Guillemette, Thomas; Simoneau, Philippe

2015-06-18

Glutathione transferases (GSTs) represent an extended family of multifunctional proteins involved in detoxification processes and tolerance to oxidative stress. We thus anticipated that some GSTs could play an essential role in the protection of fungal necrotrophs against plant-derived toxic metabolites and reactive oxygen species that accumulate at the host-pathogen interface during infection. Mining the genome of the necrotrophic Brassica pathogen Alternaria brassicicola for glutathione transferase revealed 23 sequences, 17 of which could be clustered into the main classes previously defined for fungal GSTs and six were 'orphans'. Five isothiocyanate-inducible GSTs from five different classes were more thoroughly investigated. Analysis of their catalytic properties revealed that two GSTs, belonging to the GSTFuA and GTT1 classes, exhibited GSH transferase activity with isothiocyanates (ITC) and peroxidase activity with cumene hydroperoxide, respectively. Mutant deficient for these two GSTs were however neither more susceptible to ITC nor less aggressive than the wild-type parental strain. By contrast mutants deficient for two other GSTs, belonging to the Ure2pB and GSTO classes, were distinguished by their hyper-susceptibility to ITC and low aggressiveness against Brassica oleracea. In particular AbGSTO1 could participate in cell tolerance to ITC due to its glutathione-dependent thioltransferase activity. The fifth ITC-inducible GST belonged to the MAPEG class and although it was not possible to produce the soluble active form of this protein in a bacterial expression system, the corresponding deficient mutant failed to develop normal symptoms on host plant tissues. Among the five ITC-inducible GSTs analyzed in this study, three were found essential for full aggressiveness of A. brassicicola on host plant. This, to our knowledge is the first evidence that GSTs might be essential virulence factors for fungal necrotrophs.

Degradation products of citrus volatile organic compounds (VOCs) acting as phagostimulants that increase probing behavior of Asian citrus psyllid

USDA-ARS?s Scientific Manuscript database

Volatile phytochemicals play a role in orientation by phytophagous insects. We studied antennal and behavioral responses of the Asian citrus psyllid, Diaphorina citri Kuwayama, vector of the citrus greening disease pathogen. Little or no response to citrus leaf volatiles was detected by electroanten...

Characterization and Pathogenicity of Alternaria burnsii from Seeds of Cucurbita maxima (Cucurbitaceae) in Bangladesh.

PubMed

Paul, Narayan Chandra; Deng, Jian Xin; Lee, Hyang Burm; Yu, Seung-Hun

2015-12-01

In the course of survey of endophytic fungi from Bangladesh pumpkin seeds in 2011~2012, two strains (CNU111042 and CNU111043) with similar colony characteristics were isolated and characterized by their morphology and by molecular phylogenetic analysis of the internal transcribed spacer, glyceraldehydes-3-phosphate dehydrogenase (gpd), and Alternaria allergen a1 (Alt a1) sequences. Phylogenetic analysis of all three sequences and their combined dataset revealed that the fungus formed a subclade within the A. alternata clade, matching A. burnsi and showing differences with its other closely related Alternaria species, such as A. longipes, A. tomato, and A. tomaticola. Long ellipsoid, obclavate or ovoid beakless conidia, shorter and thinner conidial size (16~60 [90] × 6.5~14 [~16] µm) distinguish this fungus from other related species. These isolates showed more transverse septation (2~11) and less longitudinal septation (0~3) than did other related species. Moreover, the isolate did not produce any diffusible pigment on media. Therefore, our results reveal that the newly recorded fungus from a new host, Cucurbita maxima, is Alternaria burnsii Uppal, Patel & Kamat.

Hyperspectral and thermal imaging of oilseed rape (Brassica napus) response to fungal species of the genus Alternaria.

PubMed

Baranowski, Piotr; Jedryczka, Malgorzata; Mazurek, Wojciech; Babula-Skowronska, Danuta; Siedliska, Anna; Kaczmarek, Joanna

2015-01-01

In this paper, thermal (8-13 µm) and hyperspectral imaging in visible and near infrared (VNIR) and short wavelength infrared (SWIR) ranges were used to elaborate a method of early detection of biotic stresses caused by fungal species belonging to the genus Alternaria that were host (Alternaria alternata, Alternaria brassicae, and Alternaria brassicicola) and non-host (Alternaria dauci) pathogens to oilseed rape (Brassica napus L.). The measurements of disease severity for chosen dates after inoculation were compared to temperature distributions on infected leaves and to averaged reflectance characteristics. Statistical analysis revealed that leaf temperature distributions on particular days after inoculation and respective spectral characteristics, especially in the SWIR range (1000-2500 nm), significantly differed for the leaves inoculated with A. dauci from the other species of Alternaria as well as from leaves of non-treated plants. The significant differences in leaf temperature of the studied Alternaria species were observed in various stages of infection development. The classification experiments were performed on the hyperspectral data of the leaf surfaces to distinguish days after inoculation and Alternaria species. The second-derivative transformation of the spectral data together with back-propagation neural networks (BNNs) appeared to be the best combination for classification of days after inoculation (prediction accuracy 90.5%) and Alternaria species (prediction accuracy 80.5%).

Reference Genes for Accurate Transcript Normalization in Citrus Genotypes under Different Experimental Conditions

PubMed Central

Mafra, Valéria; Kubo, Karen S.; Alves-Ferreira, Marcio; Ribeiro-Alves, Marcelo; Stuart, Rodrigo M.; Boava, Leonardo P.; Rodrigues, Carolina M.; Machado, Marcos A.

2012-01-01

Real-time reverse transcription PCR (RT-qPCR) has emerged as an accurate and widely used technique for expression profiling of selected genes. However, obtaining reliable measurements depends on the selection of appropriate reference genes for gene expression normalization. The aim of this work was to assess the expression stability of 15 candidate genes to determine which set of reference genes is best suited for transcript normalization in citrus in different tissues and organs and leaves challenged with five pathogens (Alternaria alternata, Phytophthora parasitica, Xylella fastidiosa and Candidatus Liberibacter asiaticus). We tested traditional genes used for transcript normalization in citrus and orthologs of Arabidopsis thaliana genes described as superior reference genes based on transcriptome data. geNorm and NormFinder algorithms were used to find the best reference genes to normalize all samples and conditions tested. Additionally, each biotic stress was individually analyzed by geNorm. In general, FBOX (encoding a member of the F-box family) and GAPC2 (GAPDH) was the most stable candidate gene set assessed under the different conditions and subsets tested, while CYP (cyclophilin), TUB (tubulin) and CtP (cathepsin) were the least stably expressed genes found. Validation of the best suitable reference genes for normalizing the expression level of the WRKY70 transcription factor in leaves infected with Candidatus Liberibacter asiaticus showed that arbitrary use of reference genes without previous testing could lead to misinterpretation of data. Our results revealed FBOX, SAND (a SAND family protein), GAPC2 and UPL7 (ubiquitin protein ligase 7) to be superior reference genes, and we recommend their use in studies of gene expression in citrus species and relatives. This work constitutes the first systematic analysis for the selection of superior reference genes for transcript normalization in different citrus organs and under biotic stress. PMID:22347455

New antitumour fungal metabolites from Alternaria porri.

PubMed

Phuwapraisirisan, Preecha; Rangsan, Jakaphan; Siripong, Pongpan; Tip-Pyang, Santi

2009-01-01

Chemical investigation of the onion pathogenic fungus Alternaria porri resulted in the isolation of two new phthalides named zinnimide (2) and deprenylzinnimide (8), along with a new bianthraquinone, alterporriol F (10). The structures of the new metabolites were characterised by spectroscopic analysis and chemical degradation. Of the new compounds isolated, alterporriol F was highly cytotoxic towards HeLa and KB cells, with IC(50) values of 6.5 and 7.0 microg mL(-1).

Hyperspectral and Thermal Imaging of Oilseed Rape (Brassica napus) Response to Fungal Species of the Genus Alternaria

PubMed Central

Baranowski, Piotr; Jedryczka, Malgorzata; Mazurek, Wojciech; Babula-Skowronska, Danuta; Siedliska, Anna; Kaczmarek, Joanna

2015-01-01

In this paper, thermal (8-13 µm) and hyperspectral imaging in visible and near infrared (VNIR) and short wavelength infrared (SWIR) ranges were used to elaborate a method of early detection of biotic stresses caused by fungal species belonging to the genus Alternaria that were host (Alternaria alternata, Alternaria brassicae, and Alternaria brassicicola) and non-host (Alternaria dauci) pathogens to oilseed rape (Brassica napus L.). The measurements of disease severity for chosen dates after inoculation were compared to temperature distributions on infected leaves and to averaged reflectance characteristics. Statistical analysis revealed that leaf temperature distributions on particular days after inoculation and respective spectral characteristics, especially in the SWIR range (1000-2500 nm), significantly differed for the leaves inoculated with A. dauci from the other species of Alternaria as well as from leaves of non-treated plants. The significant differences in leaf temperature of the studied Alternaria species were observed in various stages of infection development. The classification experiments were performed on the hyperspectral data of the leaf surfaces to distinguish days after inoculation and Alternaria species. The second-derivative transformation of the spectral data together with back-propagation neural networks (BNNs) appeared to be the best combination for classification of days after inoculation (prediction accuracy 90.5%) and Alternaria species (prediction accuracy 80.5%). PMID:25826369

Horsfall-Barratt recalibration and replicated severity estimates of citrus canker

USDA-ARS?s Scientific Manuscript database

Citrus canker is a serious disease of citrus in tropical and subtropical citrus growing regions. Accurate and precise assessment of citrus canker and other plant pathogens is needed to obtain good quality data. Citrus canker assessment data were used to ascertain some of the mechanics of the Horsfal...

The citrus postharvest pathogen Penicillium digitatum depends on the PdMpkB kinase for developmental and virulence functions.

PubMed

Ma, Haijie; Sun, Xuepeng; Wang, Mingshuang; Gai, Yunpeng; Chung, Kuang-Ren; Li, Hongye

2016-11-07

The postharvest pathogen Penicillium digitatum causes green mold decay on citrus fruit, resulting in severe economic losses. To explore possible factors involved in fungal pathogenesis, phenotypic characterization of the budding yeast Fus3/Kiss1 mitogen-activated protein (MAP) kinase homolog was carried out. The P. digitatum MAP kinase B coding gene, designated PdMpkB, was functionally inactivated via homologous recombination. The fungal strain (∆PdMpkB) carrying a PdMpkBdeletion demonstrated altered gene expression profiles, reduced growth and conidiogenesis, elevated resistance to osmotic stress, and failed to induce green mold decay on citrus fruit. ∆PdMpkB was more resistant to CaCl2, NaCl and sorbitol than its progenitor strain, indicating a negative regulatory function of PdMpkB in osmotic stress adaptation. Fungal infection assays on citrus fruit revealed that ∆PdMpkB proliferated poorly within host tissues, induced water-soaking lesions, failed to break through host cuticle layers and thus, failed to produce aerial hyphae and conidia. Introduction of a functional copy of PdMpkB into a null mutant restored all defective phenotypes. Transcriptome analysis revealed that inactivation of PdMpkB impacted expression of the genes associated with cell wall-degrading enzyme activities, carbohydrate and amino acid metabolisms, conidial formation, and numerous metabolic processes. Our results define pivotal roles of the PdMpkB-mediated signaling pathway in developmental and pathological functions in the citrus postharvest pathogen P. digitatum. Copyright © 2016 Elsevier B.V. All rights reserved.

Characterization of a novel single-stranded RNA virus, closely related to fusariviruses, infecting the plant pathogenic fungus Alternaria brassicicola.

PubMed

Zhong, Jie; Shang, Hong Hong; Zhu, Chuan Xia; Zhu, Jun Zi; Zhu, Hong Jian; Hu, Yan; Gao, Bi Da

2016-06-02

The alternaria blackspot of rapeseed is one of the most prominent diseases of rapeseed. It is caused by three species of the genus Alternaria: Alternaria brassicicola, Alternaria brassicae, and Alternaria raphanin. Here we report a novel positive-sense RNA virus from an A. brassicicola strain 817-14. The virus has a 6639 nucleotide (nt) long genome, excluding a poly (A)-tail, and was predicted to contain three putative open reading frames (ORF1, ORF2, and ORF3). The large ORF1 encoded a 174-kDa polyprotein (composed of 1522 amino acid residues) containing a conserved RNA-dependent RNA polymerase (RdRp) domain and a helicase domain. The other two smaller ORFs encoded polypeptides with unknown function. Homology search and phylogenetic analysis, based on the RdRp and helicase domains, suggest that this virus is related to and grouped with Sclerotinia sclerotiorum fusarivirus 1 (SsFV1), Rosellinia necatrix fusarivirus 1 (RnFV1), Fusarium graminearum virus-DK21 (FgV1), and Penicillium roqueforti RNA mycovirus 1 (PrRV1), all of which belong to a newly proposed family Fusariviridae. For this study, we designed the virus as "Alternaria brassicicola fusarivirus 1" (AbFV1). Virus elimination revealed that AbFV1 has no conspicuous impact on the biological properties of its host. Copyright © 2016. Published by Elsevier B.V.

Functional characterization of the Dsc E3 ligase complex in the citrus postharvest pathogen Penicillium digitatum.

PubMed

Ruan, Ruoxin; Chung, Kuang-Ren; Li, Hongye

2017-12-01

Sterol regulatory element binding proteins (SREBPs) are required for sterol homeostasis in eukaryotes. Activation of SREBPs is regulated by the Dsc E3 ligase complex in Schizosaccharomyces pombe and Aspergillus spp. Previous studies indicated that an SREBP-coding gene PdsreA is required for fungicide resistance and ergosterol biosynthesis in the citrus postharvest pathogen Penicillium digitatum. In this study, five genes, designated PddscA, PddscB, PddscC, PddscD, and PddscE encoding the Dsc E3 ligase complex were characterized to be required for fungicide resistance, ergosterol biosynthesis and CoCl 2 tolerance in P. digitatum. Each of the dsc genes was inactivated by target gene disruption and the resulted phenotypes were analyzed and compared. Genetic analysis reveals that, of five Dsc complex components, PddscB is the core subunit gene in P. digitatum. Although the resultant dsc mutants were able to infect citrus fruit and induce maceration lesions as the wild-type, the mutants rarely produced aerial mycelia on affected citrus fruit peels. P. digitatum Dsc proteins regulated not only the expression of genes involved in ergosterol biosynthesis but also that of PdsreA. Yeast two-hybrid assays revealed a direct interaction between the PdSreA protein and the Dsc proteins. Ectopic expression of the PdSreA N-terminus restored fungicide resistance in the dsc mutants. Our results provide important evidence to understand the mechanisms underlying SREBP activation and regulation of ergosterol biosynthesis in plant pathogenic fungi. Copyright © 2017 Elsevier GmbH. All rights reserved.

Additive roles of PthAs in bacterial growth and pathogenicity associated with nucleotide polymorphisms in effector-binding elements of citrus canker susceptibility genes.

PubMed

Abe, Valeria Yukari; Benedetti, Celso Eduardo

2016-10-01

Citrus canker, caused by Xanthomonas citri, affects most commercial citrus varieties. All X. citri strains possess at least one transcription activator-like effector of the PthA family that activates host disease susceptibility (S) genes. The X. citri strain 306 encodes four PthA effectors; nevertheless, only PthA4 is known to elicit cankers on citrus. As none of the PthAs act as avirulence factors on citrus, we hypothesized that PthAs 1-3 might also contribute to pathogenicity on certain hosts. Here, we show that, although PthA4 is indispensable for canker formation in six Brazilian citrus varieties, PthAs 1 and 3 contribute to canker development in 'Pera' sweet orange, but not in 'Tahiti' lemon. Deletions in two or more pthA genes reduce bacterial growth in planta more pronouncedly than single deletions, suggesting an additive role of PthAs in pathogenicity and bacterial fitness. The contribution of PthAs 1 and 3 in canker formation in 'Pera' plants does not correlate with the activation of the canker S gene, LOB1 (LATERAL ORGAN BOUNDARIES 1), but with the induction of other PthA targets, including LOB2 and citrus dioxygenase (DIOX). LOB1, LOB2 and DIOX show differential PthA-dependent expression between 'Pera' and 'Tahiti' plants that appears to be associated with nucleotide polymorphisms found at or near PthA-binding sites. We also present evidence that LOB1 activation alone is not sufficient to elicit cankers on citrus, and that DIOX acts as a canker S gene in 'Pera', but not 'Tahiti', plants. Our results suggest that the activation of multiple S genes, such as LOB1 and DIOX, is necessary for full canker development. © 2015 BSPP and John Wiley & Sons Ltd.

Genome Sequence of the Necrotrophic Plant Pathogen Alternaria brassicicola Abra43

PubMed Central

Belmas, Elodie; Briand, Martial; Kwasiborski, Anthony; Colou, Justine; N’Guyen, Guillaume; Iacomi, Béatrice; Grappin, Philippe; Campion, Claire; Simoneau, Philippe; Barret, Matthieu

2018-01-01

ABSTRACT Alternaria brassicicola causes dark spot (or black spot) disease, which is one of the most common and destructive fungal diseases of Brassicaceae spp. worldwide. Here, we report the draft genome sequence of strain Abra43. The assembly comprises 29 scaffolds, with an N50 value of 2.1 Mb. The assembled genome was 31,036,461 bp in length, with a G+C content of 50.85%. PMID:29439047

Optimal strategies for the eradication of Asiatic citrus canker in heterogeneous host landscapes

USDA-ARS?s Scientific Manuscript database

The eradication of non-native plant pathogens is a key challenge in plant disease epidemiology. Asiatic citrus canker is an economically significant disease of citrus caused by the bacterial plant pathogen Xanthomonas citri subsp. citri. The pathogen is a major exotic disease problem in many citru...

Citrus Functional Genomics and Molecular Modeling in Relation to Citrus sinensis (Sweet Orange) Infection with Xylella fastidiosa (Citrus Variegated Chlorosis).

PubMed

Dwivedi, Upendra N; Tiwari, Sameeksha; Prasanna, Pragya; Awasthi, Manika; Singh, Swati; Pandey, Veda P

2016-08-01

Citrus are among the economically most important fruit tree crops in the world. Citrus variegated chlorosis (CVC), caused by Xylella fastidiosa infection, is a serious disease limiting citrus production at a global scale. With availability of citrus genomic resources, it is now possible to compare citrus expressed sequence tag (EST) data sets and identify single-nucleotide polymorphisms (SNPs) within and among different citrus cultivars that can be exploited for citrus resistance to infections, citrus breeding, among others. We report here, for the first time, SNPs in the EST data sets of X. fastidiosa-infected Citrus sinensis (sweet orange) and their functional annotation that revealed the involvement of eight C. sinensis candidate genes in CVC pathogenesis. Among these genes were xyloglucan endotransglycosylase, myo-inositol-1-phosphate synthase, and peroxidase were found to be involved in plant cell wall metabolism. These have been further investigated by molecular modeling for their role in CVC infection and defense. Molecular docking analyses of the wild and the mutant (SNP containing) types of the selected three enzymes with their respective substrates revealed a significant decrease in the binding affinity of substrates for the mutant enzymes, thus suggesting a decrease in the catalytic efficiency of these enzymes during infection, thereby facilitating a favorable condition for infection by the pathogen. These findings offer novel agrigenomics insights in developing future molecular targets and strategies for citrus fruit cultivation in ways that are resistant to X. fastidiosa infection, and by extension, with greater harvesting efficiency and economic value.

Metabolic interplay between the Asian citrus psyllid and its Profftella symbiont: An Achilles’ heel of the citrus greening insect vector

USDA-ARS?s Scientific Manuscript database

‘Candidatus Liberibacter asiaticus’ (CLas), the bacterial pathogen associated with citrus greening disease, is transmitted by Diaphorina citri, the Asian citrus psyllid. Interactions among D. citri and its microbial endosymbionts, including ‘Candidatus Profftella armatura’, are likely to impact tra...

Annotation of the Asian citrus psyllid genome reveals a reduced innate immune system

USDA-ARS?s Scientific Manuscript database

Citrus production worldwide is currently facing significant losses due to citrus greening disease, also known as huanglongbing. The citrus greening bacteria, Candidatus Liberibacter asiaticus (CLas), is a persistent propagative pathogen transmitted by the Asian citrus psyllid, Diaphorina citri Kuway...

Overexpression of a citrus NDR1 ortholog increases disease resistance in Arabidopsis.

PubMed

Lu, Hua; Zhang, Chong; Albrecht, Ute; Shimizu, Rena; Wang, Guanfeng; Bowman, Kim D

2013-01-01

Emerging devastating diseases, such as Huanglongbing (HLB) and citrus canker, have caused tremendous losses to the citrus industry worldwide. Genetic engineering is a powerful approach that could allow us to increase citrus resistance against these diseases. The key to the success of this approach relies on a thorough understanding of defense mechanisms of citrus. Studies of Arabidopsis and other plants have provided a framework for us to better understand defense mechanisms of citrus. Salicylic acid (SA) is a key signaling molecule involved in basal defense and resistance (R) gene-mediated defense against broad-spectrum pathogens. The Arabidopsis gene NDR1 (NON-RACE-SPECIFIC DISEASE RESISTANCE 1) is a positive regulator of SA accumulation and is specifically required for signaling mediated by a subset of R genes upon recognition of their cognate pathogen effectors. Our bioinformatic analysis identified an ortholog of NDR1 from citrus, CsNDR1. Overexpression of CsNDR1 complemented susceptibility conferred by the Arabidopsis ndr1-1 mutant to Pseudomonas syringae strains and also led to enhanced resistance to an oomycete pathogen Hyaloperonospora arabidopsidis. Such heightened resistance is associated with increased SA production and expression of the defense marker gene PATHOGENESIS RELATED 1 (PR1). In addition, we found that expression of PR1 and accumulation of SA were induced to modest levels in citrus infected with Candidatus Liberibacter asiaticus, the bacterial pathogen associated with HLB disease. Thus, our data suggest that CsNDR1 is a functional ortholog of Arabidopsis NDR1. Since Ca. L. asiaticus infection only activates modest levels of defense responses in citrus, we propose that genetically increasing SA/NDR1-mediated pathways could potentially lead to enhanced resistance against HLB, citrus canker, and other destructive diseases challenging global citrus production.

Overexpression of a citrus NDR1 ortholog increases disease resistance in Arabidopsis

PubMed Central

Lu, Hua; Zhang, Chong; Albrecht, Ute; Shimizu, Rena; Wang, Guanfeng; Bowman, Kim D.

2013-01-01

Emerging devastating diseases, such as Huanglongbing (HLB) and citrus canker, have caused tremendous losses to the citrus industry worldwide. Genetic engineering is a powerful approach that could allow us to increase citrus resistance against these diseases. The key to the success of this approach relies on a thorough understanding of defense mechanisms of citrus. Studies of Arabidopsis and other plants have provided a framework for us to better understand defense mechanisms of citrus. Salicylic acid (SA) is a key signaling molecule involved in basal defense and resistance (R) gene-mediated defense against broad-spectrum pathogens. The Arabidopsis gene NDR1 (NON-RACE-SPECIFIC DISEASE RESISTANCE 1) is a positive regulator of SA accumulation and is specifically required for signaling mediated by a subset of R genes upon recognition of their cognate pathogen effectors. Our bioinformatic analysis identified an ortholog of NDR1 from citrus, CsNDR1. Overexpression of CsNDR1 complemented susceptibility conferred by the Arabidopsis ndr1-1 mutant to Pseudomonas syringae strains and also led to enhanced resistance to an oomycete pathogen Hyaloperonospora arabidopsidis. Such heightened resistance is associated with increased SA production and expression of the defense marker gene PATHOGENESIS RELATED 1 (PR1). In addition, we found that expression of PR1 and accumulation of SA were induced to modest levels in citrus infected with Candidatus Liberibacter asiaticus, the bacterial pathogen associated with HLB disease. Thus, our data suggest that CsNDR1 is a functional ortholog of Arabidopsis NDR1. Since Ca. L. asiaticus infection only activates modest levels of defense responses in citrus, we propose that genetically increasing SA/NDR1-mediated pathways could potentially lead to enhanced resistance against HLB, citrus canker, and other destructive diseases challenging global citrus production. PMID:23761797

Ontogenic variation in citrus flush shoots and its relation with host plant finding and acceptance by Asian citrus psyllid (Hemiptera: Psyllidae)

USDA-ARS?s Scientific Manuscript database

The Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae) is a destructive insect mainly because it vectors the bacterial pathogens that cause the deadly and incurable citrus greening disease. Diaphorina citri adult females lay eggs and immature development occurs exclusively on new flush sh...

First Report of Black Spot Disease Caused by Alternaria alternata on Sweet Persimmon Fruits

PubMed Central

Lee, Jung Han; Kim, Jinwoo

2013-01-01

Black spot of sweet persimmon, caused by Alternaria alternata, occurred in an orchard in Gyeongnam province, Korea in 2012. The symptom was appearance of 0.5 to 4 cm black spots on the surface of fruit. The pathogen was isolated from flesh of disease lesions. The causal agent was identified as A. alternata by morphological characteristics and sequencers of the internal transcribed spacer (ITS) 1 and ITS4 regions of rRNA. Artificial inoculation of the pathogen resulted in development of disease symptoms and the re-isolated pathogen showed characteristics of A. alternata. PMID:24198674

Development of consumer-friendly transgenic citrus plants with potential broad spectrum resistance to HLB, Citrus canker, Phytopthora and other exotic diseases

USDA-ARS?s Scientific Manuscript database

The second year of this CRB funded project has started, which is focused on the development of citrus cultivars that exhibit disease resistance to multiple pathogens such as HLB, Phytophthora and citrus canker diseases. We are using precise genetic engineering to introduce into disease susceptible ...

Comparison of antifungal activities of Vietnamese citrus essential oils.

PubMed

Van Hung, Pham; Chi, Pham Thi Lan; Phi, Nguyen Thi Lan

2013-03-01

Citrus essential oils (EOs) are volatile compounds from citrus peels and widely used in perfumes, cosmetics, soaps and aromatherapy. In this study, inhibition of citrus EOs extracted from Vietnamese orange (Citrus sinensis), mandarin (Citrus reticulata Blanco), pomelo (Citrus grandis Osbeck) and lime (Citrus aurantifolia Swingle) on the growth of plant pathogenic fungi, Mucor hiemalis, Penicillium expansum and Fusarium proliferatum was investigated. The EOs of the citrus peels were obtained by cold-pressing method and the antifungal activity of EOs was evaluated using the agar dilution method. The results show that the EOs had significant antifungal activity. Lime EO was the best inhibitor of M. hiemalis and F. proliferatum while pomelo EO was the most effective against P. expansum. These results indicate that citrus EOs can be used as antifungal natural products in the food, pharmaceutical and cosmetic industries.

Preinfection of citrus by RB strains of Citrus tristeza virus (CTV) negatively affected expression of a modified T36 CTV vector

USDA-ARS?s Scientific Manuscript database

Genetically modified T36 Citrus tristeza virus (T36-mCTV) is showing promise in Florida to mitigate huanglongbing (HLB) by expressing antimicrobial peptides and RNAi against the presumed pathogen, “Candidatus Liberibacter asiaticus” (CLas), and its vector, the Asian citrus psyllid (ACP). To this end...

A phagostimulant blend for the Asian citrus psyllid

USDA-ARS?s Scientific Manuscript database

Chemical cues that condition orientation by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), are of great interest because it is the primary vector of the causal pathogen of citrus greening disease. Previous work in our lab identified a blend of formic and acetic acids as s...

Ectopic Expression of Xylella fastidiosa rpfF Conferring Production of Diffusible Signal Factor in Transgenic Tobacco and Citrus Alters Pathogen Behavior and Reduces Disease Severity.

PubMed

Caserta, R; Souza-Neto, R R; Takita, M A; Lindow, S E; De Souza, A A

2017-11-01

The pathogenicity of Xylella fastidiosa is associated with its ability to colonize the xylem of host plants. Expression of genes contributing to xylem colonization are suppressed, while those necessary for insect vector acquisition are increased with increasing concentrations of diffusible signal factor (DSF), whose production is dependent on RpfF. We previously demonstrated that transgenic citrus plants ectopically expressing rpfF from a citrus strain of X. fastidiosa subsp. pauca exhibited less susceptibility to Xanthomonas citri subsp. citri, another pathogen whose virulence is modulated by DSF accumulation. Here, we demonstrate that ectopic expression of rpfF in both transgenic tobacco and sweet orange also confers a reduction in disease severity incited by X. fastidiosa and reduces its colonization of those plants. Decreased disease severity in the transgenic plants was generally associated with increased expression of genes conferring adhesiveness to the pathogen and decreased expression of genes necessary for active motility, accounting for the reduced population sizes achieved in the plants, apparently by limiting pathogen dispersal through the plant. Plant-derived DSF signal molecules in a host plant can, therefore, be exploited to interfere with more than one pathogen whose virulence is controlled by DSF signaling.

Processess involved in the dispersal of Xanthomonas citri pv. citri from canker-infected citrus canopies, and in the infection of citrus foliage

USDA-ARS?s Scientific Manuscript database

Citrus canker (Xanthomonas citri subsp. citri, Xcc) is now considered endemic in Florida, and epidemics result in yield loss and market penalties both in Florida and elsewhere, where the pathogen occurs and susceptible citrus is cultivated. The bacterium is dispersed in rain splash, and storms with...

Processes involved in the dispersal of Xanthomonas citri pv. citri from canker-infectd citrus canopies, and in the infection of citrus foliage

USDA-ARS?s Scientific Manuscript database

Citrus canker (Xanthomonas citri subsp. citri, Xcc) is now considered endemic in Florida, and epidemics result in yield loss and market penalties both in Florida, and elsewhere where the pathogen occurs, and susceptible citrus is cultivated. The bacterium is dispersed in rain splash, and storms wit...

Biotransformations of 6',7'-dihydroxybergamottin and 6',7'-epoxybergamottin by the citrus-pathogenic fungi diminish cytochrome P450 3A4 inhibitory activity

USDA-ARS?s Scientific Manuscript database

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)...

Identification, characterization and mycotoxigenic ability of Alternaria spp. causing core rot of apple fruit in Greece.

PubMed

Ntasiou, Panagiota; Myresiotis, Charalampos; Konstantinou, Sotiris; Papadopoulou-Mourkidou, Euphemia; Karaoglanidis, George S

2015-03-16

Alternaria core rot is a major postharvest disease of apple fruit in several countries of the world, including Greece. The study was conducted aiming to identify the disease causal agents at species level, investigate the aggressiveness of Alternaria spp. isolates and the susceptibility of different apple varieties and determine the mycotoxigenic potential of Alternaria spp. isolates from apple fruit. Seventy-five Alternaria spp. isolates obtained from apple fruit showing core rot symptoms were identified as either Alternaria tenuissima or Alternaria arborescens at frequencies of 89.3 and 11.7%, respectively, based on the sequence of endopolygalacturonase (EndoPG) gene. Artificial inoculations of fruit of 4 different varieties (Fuji, Golden Delicious, Granny Smith and Red Delicious) and incubation at two different temperatures (2 and 25°C) showed that fruit of Fuji variety were the most susceptible and fruit of Golden Delicious the most resistant to both pathogens. In addition, the production of 3 mycotoxins, alternariol (AOH), alternariol monomethyl ether (AME) and tentoxin (TEN) was investigated in 30 isolates of both species. Mycotoxin determination was conducted both in vitro, on artificial nutrient medium and in vivo on artificially inoculated apple fruit, using a high performance liquid chromatography with diode array detector (HPLC-DAD). The results showed that most of the isolates of both species were able to produce all the 3 metabolites both in vivo and in vitro. On apple fruit A. tenuissima isolates produced more AOH than A. arborescens isolates, whereas the latter produced more TEN than the former. Such results indicate that Alternaria core rot represents a major threat of apple fruit production not only due to quantitative yield losses but also for qualitative deterioration of apple by-products. Copyright © 2014 Elsevier B.V. All rights reserved.

An attract-and-kill strategy for Asian citrus psyllid

USDA-ARS?s Scientific Manuscript database

Asian citrus psyllids (ACP) transmit the pathogen responsible for citrus greening disease. Psyllids use color, smell, taste and vibrational cues to identify their host plants and conspecifics. The main goal of this project is to develop an attract-and-kill device strategy that will exploit the psyll...

Volatile Organic Compound (VOC) profiling of Citrus tristeza virus (CTV) infection in sweet orange citrus varietals using thermal desorption gas chromatography time of flight mass spectrometry (TD-GC/TOF-MS)

USDA-ARS?s Scientific Manuscript database

Citrus tristeza virus (CTV) is a plant pathogen which predominately infects economically important citrus crops such as sweet orange, clementine, lime and grapefruit varietals. Within the last 70 years, an estimated 100 million citrus trees on sour orange rootstock have been destroyed due to CTV inf...

Improved annotation of the insect vector of citrus greening disease: Biocuration by a diverse genomics community

USDA-ARS?s Scientific Manuscript database

The Asian citrus psyllid (Diaphorina citri Kuwayama) is the insect vector of the bacterium Candidatus Liberibacter asiaticus (CLas), the pathogen associated with citrus Huanglongbing (HLB, citrus greening). HLB threatens citrus production worldwide. Suppression or reduction of the insect vector usin...

Mycotoxin production in liquid culture and on plants infected with Alternaria spp. isolated from rocket and cabbage.

PubMed

Siciliano, Ilenia; Ortu, Giuseppe; Gilardi, Giovanna; Gullino, Maria Lodovica; Garibaldi, Angelo

2015-03-05

Fungi belonging to the genus Alternaria are common pathogens of fruit and vegetables with some species able to produce secondary metabolites dangerous to human health. Twenty-eight Alternaria isolates from rocket and cabbage were investigated for their mycotoxin production. Five different Alternaria toxins were extracted from synthetic liquid media and from plant material (cabbage, cultivated rocket, cauliflower). A modified Czapek-Dox medium was used for the in vitro assay. Under these conditions, more than 80% of the isolates showed the ability to produce at least one mycotoxin, generally with higher levels for tenuazonic acid. However, the same isolates analyzed in vivo seemed to lose their ability to produce tenuazonic acid. For the other mycotoxins; alternariol, alternariol monomethyl ether, altenuene and tentoxin a good correlation between in vitro and in vivo production was observed. In vitro assay is a useful tool to predict the possible mycotoxin contamination under field and greenhouse conditions.

Protein interaction networks at the host-microbe interface in Diaphorina citri, the insect vector of the citrus greening pathogen.

PubMed

Ramsey, J S; Chavez, J D; Johnson, R; Hosseinzadeh, S; Mahoney, J E; Mohr, J P; Robison, F; Zhong, X; Hall, D G; MacCoss, M; Bruce, J; Cilia, M

2017-02-01

The Asian citrus psyllid ( Diaphorina citri) is the insect vector responsible for the worldwide spread of ' Candidatus Liberibacter asiaticus' (CLas), the bacterial pathogen associated with citrus greening disease. Developmental changes in the insect vector impact pathogen transmission, such that D. citri transmission of CLas is more efficient when bacteria are acquired by nymphs when compared with adults. We hypothesize that expression changes in the D. citri immune system and commensal microbiota occur during development and regulate vector competency. In support of this hypothesis, more proteins, with greater fold changes, were differentially expressed in response to CLas in adults when compared with nymphs, including insect proteins involved in bacterial adhesion and immunity. Compared with nymphs, adult insects had a higher titre of CLas and the bacterial endosymbionts Wolbachia, Profftella and Carsonella. All Wolbachia and Profftella proteins differentially expressed between nymphs and adults are upregulated in adults, while most differentially expressed Carsonella proteins are upregulated in nymphs. Discovery of protein interaction networks has broad applicability to the study of host-microbe relationships. Using protein interaction reporter technology, a D. citri haemocyanin protein highly upregulated in response to CLas was found to physically interact with the CLas coenzyme A (CoA) biosynthesis enzyme phosphopantothenoylcysteine synthetase/decarboxylase. CLas pantothenate kinase, which catalyses the rate-limiting step of CoA biosynthesis, was found to interact with a D. citri myosin protein. Two Carsonella enzymes involved in histidine and tryptophan biosynthesis were found to physically interact with D. citri proteins. These co-evolved protein interaction networks at the host-microbe interface are highly specific targets for controlling the insect vector responsible for the spread of citrus greening.

Protein interaction networks at the host–microbe interface in Diaphorina citri, the insect vector of the citrus greening pathogen

PubMed Central

Chavez, J. D.; Johnson, R.; Hosseinzadeh, S.; Mahoney, J. E.; Mohr, J. P.; Robison, F.; Zhong, X.; Hall, D. G.; MacCoss, M.; Bruce, J.; Cilia, M.

2017-01-01

The Asian citrus psyllid (Diaphorina citri) is the insect vector responsible for the worldwide spread of ‘Candidatus Liberibacter asiaticus’ (CLas), the bacterial pathogen associated with citrus greening disease. Developmental changes in the insect vector impact pathogen transmission, such that D. citri transmission of CLas is more efficient when bacteria are acquired by nymphs when compared with adults. We hypothesize that expression changes in the D. citri immune system and commensal microbiota occur during development and regulate vector competency. In support of this hypothesis, more proteins, with greater fold changes, were differentially expressed in response to CLas in adults when compared with nymphs, including insect proteins involved in bacterial adhesion and immunity. Compared with nymphs, adult insects had a higher titre of CLas and the bacterial endosymbionts Wolbachia, Profftella and Carsonella. All Wolbachia and Profftella proteins differentially expressed between nymphs and adults are upregulated in adults, while most differentially expressed Carsonella proteins are upregulated in nymphs. Discovery of protein interaction networks has broad applicability to the study of host–microbe relationships. Using protein interaction reporter technology, a D. citri haemocyanin protein highly upregulated in response to CLas was found to physically interact with the CLas coenzyme A (CoA) biosynthesis enzyme phosphopantothenoylcysteine synthetase/decarboxylase. CLas pantothenate kinase, which catalyses the rate-limiting step of CoA biosynthesis, was found to interact with a D. citri myosin protein. Two Carsonella enzymes involved in histidine and tryptophan biosynthesis were found to physically interact with D. citri proteins. These co-evolved protein interaction networks at the host–microbe interface are highly specific targets for controlling the insect vector responsible for the spread of citrus greening. PMID:28386418

Prophage-mediated dynamics of 'Candidatus Liberibacter asiaticus' populations, the destructive bacterial pathogens of citrus huanglongbing.

PubMed

Zhou, Lijuan; Powell, Charles A; Li, Wenbin; Irey, Mike; Duan, Yongping

2013-01-01

Prophages are highly dynamic components in the bacterial genome and play an important role in intraspecies variations. There are at least two prophages in the chromosomes of Candidatus Liberibacter asiaticus' (Las) Floridian isolates. Las is both unculturable and the most prevalent species of Liberibacter pathogens that cause huanglongbing (HLB), a worldwide destructive disease of citrus. In this study, seven new prophage variants resulting from two hyper-variable regions were identified by screening clone libraries of infected citrus, periwinkle and psyllids. Among them, Types A and B share highly conserved sequences and localize within the two prophages, FP1 and FP2, respectively. Although Types B and C were abundant in all three libraries, Type A was much more abundant in the libraries from the Las-infected psyllids than from the Las-infected plants, and Type D was only identified in libraries from the infected host plants but not from the infected psyllids. Sequence analysis of these variants revealed that the variations may result from recombination and rearrangement events. Conventional PCR results using type-specific molecular markers indicated that A, B, C and D are the four most abundant types in Las-infected citrus and periwinkle. However, only three types, A, B and C are abundant in Las-infected psyllids. Typing results for Las-infected citrus field samples indicated that mixed populations of Las bacteria present in Floridian isolates, but only the Type D population was correlated with the blotchy mottle symptom. Extended cloning and sequencing of the Type D region revealed a third prophage/phage in the Las genome, which may derive from the recombination of FP1 and FP2. Dramatic variations in these prophage regions were also found among the global Las isolates. These results are the first to demonstrate the prophage/phage-mediated dynamics of Las populations in plant and insect hosts, and their correlation with insect transmission and disease development.

Metabolic Interplay between the Asian Citrus Psyllid and Its Profftella Symbiont: An Achilles’ Heel of the Citrus Greening Insect Vector

PubMed Central

Ramsey, John S.; Johnson, Richard S.; Hoki, Jason S.; Kruse, Angela; Mahoney, Jaclyn; Hilf, Mark E.; Hunter, Wayne B.; Hall, David G.; Schroeder, Frank C.; MacCoss, Michael J.; Cilia, Michelle

2015-01-01

‘Candidatus Liberibacter asiaticus’ (CLas), the bacterial pathogen associated with citrus greening disease, is transmitted by Diaphorina citri, the Asian citrus psyllid. Interactions among D. citri and its microbial endosymbionts, including ‘Candidatus Profftella armatura’, are likely to impact transmission of CLas. We used quantitative mass spectrometry to compare the proteomes of CLas(+) and CLas(-) populations of D. citri, and found that proteins involved in polyketide biosynthesis by the endosymbiont Profftella were up-regulated in CLas(+) insects. Mass spectrometry analysis of the Profftella polyketide diaphorin in D. citri metabolite extracts revealed the presence of a novel diaphorin-related polyketide and the ratio of these two polyketides was changed in CLas(+) insects. Insect proteins differentially expressed between CLas(+) and CLas(-) D. citri included defense and immunity proteins, proteins involved in energy storage and utilization, and proteins involved in endocytosis, cellular adhesion, and cytoskeletal remodeling which are associated with microbial invasion of host cells. Insight into the metabolic interdependence between the insect vector, its endosymbionts, and the citrus greening pathogen reveals novel opportunities for control of this disease, which is currently having a devastating impact on citrus production worldwide. PMID:26580079

Mycotoxin Production in Liquid Culture and on Plants Infected with Alternaria spp. Isolated from Rocket and Cabbage

PubMed Central

Siciliano, Ilenia; Ortu, Giuseppe; Gilardi, Giovanna; Gullino, Maria Lodovica; Garibaldi, Angelo

2015-01-01

Fungi belonging to the genus Alternaria are common pathogens of fruit and vegetables with some species able to produce secondary metabolites dangerous to human health. Twenty-eight Alternaria isolates from rocket and cabbage were investigated for their mycotoxin production. Five different Alternaria toxins were extracted from synthetic liquid media and from plant material (cabbage, cultivated rocket, cauliflower). A modified Czapek-Dox medium was used for the in vitro assay. Under these conditions, more than 80% of the isolates showed the ability to produce at least one mycotoxin, generally with higher levels for tenuazonic acid. However, the same isolates analyzed in vivo seemed to lose their ability to produce tenuazonic acid. For the other mycotoxins; alternariol, alternariol monomethyl ether, altenuene and tentoxin a good correlation between in vitro and in vivo production was observed. In vitro assay is a useful tool to predict the possible mycotoxin contamination under field and greenhouse conditions. PMID:25751147

Phylogeny of Alternaria fungi known to produce host-specific toxins on the basis of variation in internal transcribed spacers of ribosomal DNA.

PubMed

Kusaba, M; Tsuge, T

1995-10-01

The internal transcribed spacer regions (ITS1 and ITS2) of ribosomal DNA from Alternaria species, including seven fungi known to produce host-specific toxins, were analyzed by polymerase chain reaction-amplification and direct sequencing. Phylogenetic analysis of the sequence data by the Neighbor-joining method showed that the seven toxin-producing fungi belong to a monophyletic group together with A. alternata. In contract, A. dianthi, A. panax, A. dauci, A. bataticola, A. porri, A. sesami and A. solani, species that can be morphologically distinguished from A. alternata, could be clearly separated from A. alternata by phylogenetic of the ITS variation. These results suggest that Alternaria pathogens which produce host-specific toxins are pathogenic variants within a single variable species, A. alternata.

Ex Vivo Application of Secreted Metabolites Produced by Soil-Inhabiting Bacillus spp. Efficiently Controls Foliar Diseases Caused by Alternaria spp.

PubMed

Ali, Gul Shad; El-Sayed, Ashraf S A; Patel, Jaimin S; Green, Kari B; Ali, Mohammad; Brennan, Mary; Norman, David

2016-01-15

Bacterial biological control agents (BCAs) are largely used as live products to control plant pathogens. However, due to variable environmental and ecological factors, live BCAs usually fail to produce desirable results against foliar pathogens. In this study, we investigated the potential of cell-free culture filtrates of 12 different bacterial BCAs isolated from flower beds for controlling foliar diseases caused by Alternaria spp. In vitro studies showed that culture filtrates from two isolates belonging to Bacillus subtilis and Bacillus amyloliquefaciens displayed strong efficacy and potencies against Alternaria spp. The antimicrobial activity of the culture filtrate of these two biological control agents was effective over a wider range of pH (3.0 to 9.0) and was not affected by autoclaving or proteolysis. Comparative liquid chromatography-mass spectrometry (LC-MS) analyses showed that a complex mixture of cyclic lipopeptides, primarily of the fengycin A and fengycin B families, was significantly higher in these two BCAs than inactive Bacillus spp. Interaction studies with mixtures of culture filtrates of these two species revealed additive activity, suggesting that they produce similar products, which was confirmed by LC-tandem MS analyses. In in planta pre- and postinoculation trials, foliar application of culture filtrates of B. subtilis reduced lesion sizes and lesion frequencies caused by Alternaria alternata by 68 to 81%. Taken together, our studies suggest that instead of live bacteria, culture filtrates of B. subtilis and B. amyloliquefaciens can be applied either individually or in combination for controlling foliar diseases caused by Alternaria species. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Ex Vivo Application of Secreted Metabolites Produced by Soil-Inhabiting Bacillus spp. Efficiently Controls Foliar Diseases Caused by Alternaria spp.

PubMed Central

El-Sayed, Ashraf S. A.; Patel, Jaimin S.; Green, Kari B.; Ali, Mohammad; Brennan, Mary; Norman, David

2015-01-01

Bacterial biological control agents (BCAs) are largely used as live products to control plant pathogens. However, due to variable environmental and ecological factors, live BCAs usually fail to produce desirable results against foliar pathogens. In this study, we investigated the potential of cell-free culture filtrates of 12 different bacterial BCAs isolated from flower beds for controlling foliar diseases caused by Alternaria spp. In vitro studies showed that culture filtrates from two isolates belonging to Bacillus subtilis and Bacillus amyloliquefaciens displayed strong efficacy and potencies against Alternaria spp. The antimicrobial activity of the culture filtrate of these two biological control agents was effective over a wider range of pH (3.0 to 9.0) and was not affected by autoclaving or proteolysis. Comparative liquid chromatography-mass spectrometry (LC-MS) analyses showed that a complex mixture of cyclic lipopeptides, primarily of the fengycin A and fengycin B families, was significantly higher in these two BCAs than inactive Bacillus spp. Interaction studies with mixtures of culture filtrates of these two species revealed additive activity, suggesting that they produce similar products, which was confirmed by LC-tandem MS analyses. In in planta pre- and postinoculation trials, foliar application of culture filtrates of B. subtilis reduced lesion sizes and lesion frequencies caused by Alternaria alternata by 68 to 81%. Taken together, our studies suggest that instead of live bacteria, culture filtrates of B. subtilis and B. amyloliquefaciens can be applied either individually or in combination for controlling foliar diseases caused by Alternaria species. PMID:26519395

Swainsonine biosynthesis genes in diverse symbiotic and pathogenic fungi

USDA-ARS?s Scientific Manuscript database

Swainsonine, a cytotoxic fungal alkaloid and a potential cancer therapy drug, is produced by the insect pathogen and plant symbiont, Metarhizium robertsii, the clover pathogen Slafractonia leguminicola, locoweed symbionts belonging to Alternaria sect. Undifilum, and a recently discovered morning glo...

Prophage-Mediated Dynamics of ‘Candidatus Liberibacter asiaticus’ Populations, the Destructive Bacterial Pathogens of Citrus Huanglongbing

PubMed Central

Zhou, Lijuan; Powell, Charles A.; Li, Wenbin; Irey, Mike; Duan, Yongping

2013-01-01

Prophages are highly dynamic components in the bacterial genome and play an important role in intraspecies variations. There are at least two prophages in the chromosomes of Candidatus Liberibacter asiaticus’ (Las) Floridian isolates. Las is both unculturable and the most prevalent species of Liberibacter pathogens that cause huanglongbing (HLB), a worldwide destructive disease of citrus. In this study, seven new prophage variants resulting from two hyper-variable regions were identified by screening clone libraries of infected citrus, periwinkle and psyllids. Among them, Types A and B share highly conserved sequences and localize within the two prophages, FP1 and FP2, respectively. Although Types B and C were abundant in all three libraries, Type A was much more abundant in the libraries from the Las-infected psyllids than from the Las-infected plants, and Type D was only identified in libraries from the infected host plants but not from the infected psyllids. Sequence analysis of these variants revealed that the variations may result from recombination and rearrangement events. Conventional PCR results using type-specific molecular markers indicated that A, B, C and D are the four most abundant types in Las-infected citrus and periwinkle. However, only three types, A, B and C are abundant in Las-infected psyllids. Typing results for Las-infected citrus field samples indicated that mixed populations of Las bacteria present in Floridian isolates, but only the Type D population was correlated with the blotchy mottle symptom. Extended cloning and sequencing of the Type D region revealed a third prophage/phage in the Las genome, which may derive from the recombination of FP1 and FP2. Dramatic variations in these prophage regions were also found among the global Las isolates. These results are the first to demonstrate the prophage/phage-mediated dynamics of Las populations in plant and insect hosts, and their correlation with insect transmission and disease

Wind speed and wind-associated leaf injury affect severity of citrus canker on Swingle citrumelo

USDA-ARS?s Scientific Manuscript database

Citrus canker (caused by the bacterial pathogen Xanthomonas citri subsp. citri, Xcc) can cause severe damage to citrus. It is endemic in Florida, and occurs in other citrus growing regions. The bacterium is dispersed predominantly in rain splash. To simulate dispersal in splash, and to investigate t...

Management of Citrus Canker in Argentina, a Success Story.

PubMed

Canteros, B I; Gochez, A M; Moschini, R C

2017-10-01

Citrus canker is an important bacterial disease of citrus in several regions of the world. Strains of Xanthomonas citri type-A (Xc-A) group are the primary pathogen where citrus canker occurs. After Xc-A entered the Northeast of Argentina in 1974, the disease spread rapidly from 1977 to 1980 and then slowed down and remained moving at slow pace until 1990 when it became endemic. Citrus canker was detected in Northwest Argentina in 2002. This paper presents the main steps in the fight of the disease and the management strategies that have been used to control citrus canker at this time. We think the process might be usefull to other countries with the same situation. Results from more than 40 years of research in Northeast (NE) Argentina indicate that we are at the limit of favorable environment for the disease. The severity of citrus canker is greatly affected by the environment and El Niño Southern Oscillation (ENSO) phenomenon which causes cyclic fluctuations on the disease intensity in the NE region. Weather-based logistic regression models adjusted to quantify disease levels in field conditions showed that the environmental effect was strongly modulated by the distance from a windbreak. Production of healthy fruits in citrus canker endemic areas is possible knowing the dynamics of the disease. A voluntary Integrated Plan to Reduce the Risk of Canker has been in place since 1994 and it allows growers to export unsymptomatic, uninfested fresh fruit to countries which are free of the disease and require healthy, pathogen free fruits. The experience from Argentina can be replicated in other countries after appropriate trials.

Management of Citrus Canker in Argentina, a Success Story

PubMed Central

Canteros, B. I.; Gochez, A. M.; Moschini, R. C.

2017-01-01

Citrus canker is an important bacterial disease of citrus in several regions of the world. Strains of Xanthomonas citri type-A (Xc-A) group are the primary pathogen where citrus canker occurs. After Xc-A entered the Northeast of Argentina in 1974, the disease spread rapidly from 1977 to 1980 and then slowed down and remained moving at slow pace until 1990 when it became endemic. Citrus canker was detected in Northwest Argentina in 2002. This paper presents the main steps in the fight of the disease and the management strategies that have been used to control citrus canker at this time. We think the process might be usefull to other countries with the same situation. Results from more than 40 years of research in Northeast (NE) Argentina indicate that we are at the limit of favorable environment for the disease. The severity of citrus canker is greatly affected by the environment and El Niño Southern Oscillation (ENSO) phenomenon which causes cyclic fluctuations on the disease intensity in the NE region. Weather-based logistic regression models adjusted to quantify disease levels in field conditions showed that the environmental effect was strongly modulated by the distance from a windbreak. Production of healthy fruits in citrus canker endemic areas is possible knowing the dynamics of the disease. A voluntary Integrated Plan to Reduce the Risk of Canker has been in place since 1994 and it allows growers to export unsymptomatic, uninfested fresh fruit to countries which are free of the disease and require healthy, pathogen free fruits. The experience from Argentina can be replicated in other countries after appropriate trials. PMID:29018307

PAMPs, PRRs, effectors and R-genes associated with citrus–pathogen interactions

PubMed Central

Dalio, Ronaldo J. D.; Magalhães, Diogo M.; Rodrigues, Carolina M.; Arena, Gabriella D.; Oliveira, Tiago S.; Souza-Neto, Reinaldo R.; Picchi, Simone C.; Martins, Paula M. M.; Santos, Paulo J. C.; Maximo, Heros J.; Pacheco, Inaiara S.; De Souza, Alessandra A.

2017-01-01

Abstract Background Recent application of molecular-based technologies has considerably advanced our understanding of complex processes in plant–pathogen interactions and their key components such as PAMPs, PRRs, effectors and R-genes. To develop novel control strategies for disease prevention in citrus, it is essential to expand and consolidate our knowledge of the molecular interaction of citrus plants with their pathogens. Scope This review provides an overview of our understanding of citrus plant immunity, focusing on the molecular mechanisms involved in the interactions with viruses, bacteria, fungi, oomycetes and vectors related to the following diseases: tristeza, psorosis, citrus variegated chlorosis, citrus canker, huanglongbing, brown spot, post-bloom, anthracnose, gummosis and citrus root rot. PMID:28065920

Insecticidal Suppression of Asian Citrus Psyllid Diaphorina citri (Hemiptera: Liviidae) Vector of Huanglongbing Pathogens

PubMed Central

Qureshi, Jawwad A.; Kostyk, Barry C.; Stansly, Philip A.

2014-01-01

Diaphorina citri vectors pathogens that cause ‘huanglongbing’ or citrus greening disease which poses a serious threat to citrus production worldwide. Vector suppression is critical to reduce disease spread. Efficacy is a main concern when choosing an insecticide. Insecticidal treatments of 49 products or 44 active ingredients (a.i) labeled or experimental were field tested between 2005–2013 as foliar sprays (250 treatments, 39 a.i) or soil applications (47 treatments, 9 a.i) to control D. citri in citrus. A combined effect of nymphal and adult suppression in response to sprays of 23 insecticides representing 9 modes of action (MoA) groups and 3 unknown MoA provided more than 90% reduction of adult D. citri over 24–68 days. Observable effects on nymphs were generally of shorter duration due to rapid maturation of flush. However, reduction of 76–100% nymphs or adults over 99–296 days was seen on young trees receiving drenches of the neonicotinoids imidacloprid, thiamethoxam or clothianidin (MoA 4A) and a novel anthranilic diamide, cyantraniliprole (MoA 28). Effective products identified for foliar sprays to control D. citri provide sufficient MoA groups for rotation to delay evolution of insecticide resistance by D. citri and other pests. However, cyantraniliprole is now the only available alternative for rotation with neonicotinoids in soil application to young trees. Sprays of up to eight of the most effective insecticides could be rotated over a year without repetition of any MoA and little or no recourse to neonicotinoids or cyantraniliprole, so important for protection of young trees. Other considerations effecting decisions of what and when to spray include prevalence of huanglongbing, pest pressure, pre-harvest intervals, overall budget, equipment availability, and conservation of beneficial arthropods. Examples of spray programs utilizing broad-spectrum and relatively selective insecticides are provided to improve vector management and may vary

Insecticidal suppression of Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae) vector of huanglongbing pathogens.

PubMed

Qureshi, Jawwad A; Kostyk, Barry C; Stansly, Philip A

2014-01-01

Diaphorina citri vectors pathogens that cause 'huanglongbing' or citrus greening disease which poses a serious threat to citrus production worldwide. Vector suppression is critical to reduce disease spread. Efficacy is a main concern when choosing an insecticide. Insecticidal treatments of 49 products or 44 active ingredients (a.i) labeled or experimental were field tested between 2005-2013 as foliar sprays (250 treatments, 39 a.i) or soil applications (47 treatments, 9 a.i) to control D. citri in citrus. A combined effect of nymphal and adult suppression in response to sprays of 23 insecticides representing 9 modes of action (MoA) groups and 3 unknown MoA provided more than 90% reduction of adult D. citri over 24-68 days. Observable effects on nymphs were generally of shorter duration due to rapid maturation of flush. However, reduction of 76-100% nymphs or adults over 99-296 days was seen on young trees receiving drenches of the neonicotinoids imidacloprid, thiamethoxam or clothianidin (MoA 4A) and a novel anthranilic diamide, cyantraniliprole (MoA 28). Effective products identified for foliar sprays to control D. citri provide sufficient MoA groups for rotation to delay evolution of insecticide resistance by D. citri and other pests. However, cyantraniliprole is now the only available alternative for rotation with neonicotinoids in soil application to young trees. Sprays of up to eight of the most effective insecticides could be rotated over a year without repetition of any MoA and little or no recourse to neonicotinoids or cyantraniliprole, so important for protection of young trees. Other considerations effecting decisions of what and when to spray include prevalence of huanglongbing, pest pressure, pre-harvest intervals, overall budget, equipment availability, and conservation of beneficial arthropods. Examples of spray programs utilizing broad-spectrum and relatively selective insecticides are provided to improve vector management and may vary depending on

The quest for a non-vector psyllid: Natural variation in acquisition and transmission of the huanglongbing pathogen 'Candidatus Liberibacter asiaticus' by Asian citrus psyllid isofemale lines

USDA-ARS?s Scientific Manuscript database

Genetic variability in insect vectors is valuable to study vector competence determinants and to select non-vector populations that may help reduce the spread of vector-borne pathogens. We collected and tested vector competency of 15 isofemale lines of Asian citrus psyllid (ACP) Diaphorina citri, v...

Rapid screening for citrus canker resistance employing pattern-triggered immunity (PTI) responses

USDA-ARS?s Scientific Manuscript database

Citrus canker, caused by the bacterial pathogen Xanthomonas citri ssp. citri (Xcc), has been attributed to millions of dollars in loss or damage to commercial citrus crops in subtropical production areas of the world. Since identification of resistant plants is one of the most effective methods of d...

Survival of Alternaria alternata during anaerobic digestion of biomass in stirred tank reactors.

PubMed

Schleusner, Y; Bandte, M; Gossmann, M; Heiermann, M; Plöchl, M; Büttner, C

2012-01-01

The survival of Alternaria alternate during anaerobic digestion was investigated in context of a joint research project. The aim of this project was to estimate the phytosanitary risk of dissemination of pathogens by returning treated biomass as organic fertilizer to arable land. The studies were carried out in lab-scale stirred tank reactors under mesophilic conditions. After insertion of infected plant material into the reactors the influence on the viability of the fungal pathogen was studied concerning exposure time, pretreatment and storage of the digestates for four weeks or six months. The results clearly showed that anaerobic digestion leads to a complete inactivation of A. alternate already after an exposure time of six hours.

An Inexpensive and Safe Experiment to Demonstrate Koch's Postulates Using Citrus Fruit

ERIC Educational Resources Information Center

Jakobi, Steven

2010-01-01

Citrus fruit (oranges, tangerines, grapefruit or lemons) purchased in a grocery store can be experimentally infected with readily-available sources of "Penicillium digitatum" to demonstrate the four basic steps of Koch's postulates, also known as proof of pathogenicity. The mould is isolated from naturally-infected citrus fruit into pure culture…

Identification of putative TAL effector targets of the citrus canker pathogens shows functional convergence underlying disease development and defense response

PubMed Central

2014-01-01

Background Transcriptional activator-like (TAL) effectors, formerly known as the AvrBs3/PthA protein family, are DNA-binding effectors broadly found in Xanthomonas spp. that transactivate host genes upon injection via the bacterial type three-secretion system. Biologically relevant targets of TAL effectors, i.e. host genes whose induction is vital to establish a compatible interaction, have been reported for xanthomonads that colonize rice and pepper; however, citrus genes modulated by the TAL effectors PthA“s” and PthC“s” of the citrus canker bacteria Xanthomonas citri (Xc) and Xanthomonas aurantifolii pathotype C (XaC), respectively, are poorly characterized. Of particular interest, XaC causes canker disease in its host lemon (Citrus aurantifolia), but triggers a defense response in sweet orange. Results Based on, 1) the TAL effector-DNA binding code, 2) gene expression data of Xc and XaC-infiltrated sweet orange leaves, and 3) citrus hypocotyls transformed with PthA2, PthA4 or PthC1, we have identified a collection of Citrus sinensis genes potentially targeted by Xc and XaC TAL effectors. Our results suggest that similar with other strains of Xanthomonas TAL effectors, PthA2 and PthA4, and PthC1 to some extent, functionally converge. In particular, towards induction of genes involved in the auxin and gibberellin synthesis and response, cell division, and defense response. We also present evidence indicating that the TAL effectors act as transcriptional repressors and that the best scoring predicted DNA targets of PthA“s” and PthC“s” in citrus promoters predominantly overlap with or localize near to TATA boxes of core promoters, supporting the idea that TAL effectors interact with the host basal transcriptional machinery to recruit the RNA pol II and start transcription. Conclusions The identification of PthA“s” and PthC“s” targets, such as the LOB (LATERAL ORGAN BOUNDARY) and CCNBS genes that we report here, is key for the understanding

Identification of putative TAL effector targets of the citrus canker pathogens shows functional convergence underlying disease development and defense response.

PubMed

Pereira, Andre L A; Carazzolle, Marcelo F; Abe, Valeria Y; de Oliveira, Maria L P; Domingues, Mariane N; Silva, Jaqueline C; Cernadas, Raul A; Benedetti, Celso E

2014-02-25

Transcriptional activator-like (TAL) effectors, formerly known as the AvrBs3/PthA protein family, are DNA-binding effectors broadly found in Xanthomonas spp. that transactivate host genes upon injection via the bacterial type three-secretion system. Biologically relevant targets of TAL effectors, i.e. host genes whose induction is vital to establish a compatible interaction, have been reported for xanthomonads that colonize rice and pepper; however, citrus genes modulated by the TAL effectors PthA"s" and PthC"s" of the citrus canker bacteria Xanthomonas citri (Xc) and Xanthomonas aurantifolii pathotype C (XaC), respectively, are poorly characterized. Of particular interest, XaC causes canker disease in its host lemon (Citrus aurantifolia), but triggers a defense response in sweet orange. Based on, 1) the TAL effector-DNA binding code, 2) gene expression data of Xc and XaC-infiltrated sweet orange leaves, and 3) citrus hypocotyls transformed with PthA2, PthA4 or PthC1, we have identified a collection of Citrus sinensis genes potentially targeted by Xc and XaC TAL effectors. Our results suggest that similar with other strains of Xanthomonas TAL effectors, PthA2 and PthA4, and PthC1 to some extent, functionally converge. In particular, towards induction of genes involved in the auxin and gibberellin synthesis and response, cell division, and defense response. We also present evidence indicating that the TAL effectors act as transcriptional repressors and that the best scoring predicted DNA targets of PthA"s" and PthC"s" in citrus promoters predominantly overlap with or localize near to TATA boxes of core promoters, supporting the idea that TAL effectors interact with the host basal transcriptional machinery to recruit the RNA pol II and start transcription. The identification of PthA"s" and PthC"s" targets, such as the LOB (lateral organ boundary) and CCNBS genes that we report here, is key for the understanding of the canker symptoms development during host

Occurrence and function of fungal antifungal proteins: a case study of the citrus postharvest pathogen Penicillium digitatum.

PubMed

Garrigues, Sandra; Gandía, Mónica; Marcos, Jose F

2016-03-01

Antifungal proteins (AFPs) of fungal origin have been described in filamentous fungi. AFPs are small, highly stable, cationic cysteine-rich proteins (CRPs) that are usually secreted in high amounts and show potent antifungal activity against non-self fungi. The role of AFPs in the biology of the producer fungus remains unclear. AFPs have been proposed as promising lead compounds for the development of new antifungals. The analyses of available antifungal CRP sequences from fungal origin and their phylogenetic reconstruction led us to propose a new classification of AFPs in three distinct classes: A, B and C. We initiate for the first time the characterization of an AFP in a fungal pathogen, by analysing the functional role of the unique afpB gene in the citrus fruit pathogen Penicillium digitatum. Null ΔafpB mutants revealed that this gene is dispensable for vegetative growth and fruit infection. However, strains that artificially express afpB in a constitutive way (afpB (C)) showed a phenotype of restricted growth, distortion of hyphal morphology and strong reduction in virulence to citrus fruits. These characteristics support an antifungal role for AfpB. Surprisingly, we did not detect the AfpB protein in any of the P. digitatum strains and growth conditions that were analysed in this study, regardless of high gene expression. The afpB (C) phenotype is not stable and occasionally reverts to a wild type-like phenotype but molecular changes were not detected with this reversion. The reduced virulence of afpB (C) strains correlated with localized fruit necrosis and altered timing of expression of fruit defence genes.

Data on morphological features of mycosis induced by Colletotrichum nymphaeae and Lecanicillium longisporum on citrus orthezia scale.

PubMed

Mascarin, Gabriel Moura; Guarín-Molina, Juan Humberto; Arthurs, Steven Paul; Humber, Richard Alan; de Andrade Moral, Rafael; Demétrio, Clarice Garcia Borges; Delalibera, Ítalo

2016-09-01

We describe symptoms of mycosis induced by two native fungal entomopathogens of the citrus orthezia scale, Praelongorthezia praelonga (Hemiptera: Ortheziidae), an important pest of citrus orchards. The data presented in this article are related to the article entitled "Seasonal prevalence of the insect pathogenic fungus Colletotrichum nymphaeae in Brazilian citrus groves under different chemical pesticide regimes" [1]. The endemic fungal pathogen, C. nymphaeae, emerges through the thin cuticular intersegmental regions of the citrus orthezia scale body revealing orange salmon-pigmented conidiophores bearing conidial masses, as well as producing rhizoid-like hyphae that extend over the citrus leaf. By contrast, nymphs or adult females of this scale insect infected with Lecanicillium longisporum exhibit profuse outgrowth of bright white-pigmented conidiophores with clusters of conidia emerging from the insect intersegmental membranes, and mycosed cadavers are commonly observed attached to the leaf surface by hyphal extensions. These morphological differences are important features to discriminate these fungal entomopathogens in citrus orthezia scales.

Visual rating and the use of image analysis for assessing different symptoms of citrus canker on grapefruit leaves

USDA-ARS?s Scientific Manuscript database

Citrus canker is caused by the bacterial pathogen Xanthomonas axonopodis pv citri (Xac) and infects several citrus species in wet tropical and subtropical citrus growing regions. Accurate, precise and reproducible disease assessment is needed for monitoring epidemics and disease response in breeding...

Quantitative distribution of 'Candidatus Liberibacter asiaticus' in citrus plants with citrus huanglongbing.

PubMed

Li, Wenbin; Levy, Laurene; Hartung, John S

2009-02-01

Citrus huanglongbing (HLB), or greening disease, is strongly associated with any of three nonculturable gram-negative bacteria belonging to 'Candidatus Liberibacter spp.' 'Ca. Liberibacter spp.' are transmitted by citrus psyllids to all commercial cultivars of citrus. The diseases can be lethal to citrus and have recently become widespread in both São Paulo, Brazil, and Florida, United States, the locations of the largest citrus industries in the world. Asiatic HLB, the form of the disease found in Florida, is associated with 'Ca. Liberibacter asiaticus' and is the subject of this report. The nonculturable nature of the pathogen has hampered research and little is known about the distribution of 'Ca. L. asiaticus' in infected trees. In this study, we have used a quantitative polymerase chain reaction assay to systematically quantify the distribution of 'Ca. L. asiaticus' genomes in tissues of six species of citrus either identified in the field during survey efforts in Florida or propagated in a greenhouse in Beltsville, MD. The populations of 'Ca. L. asiaticus' inferred from the distribution of 16S rDNA sequences specific for 'Ca. L. asiaticus' in leaf midribs, leaf blades, and bark samples varied by a factor of 1,000 among samples prepared from the six citrus species tested and by a factor of 100 between two sweet orange trees tested. In naturally infected trees, above-ground portions of the tree averaged 10(10) 'Ca. L. asiaticus' genomes per gram of tissue. Similar levels of 'Ca. L. asiaticus' genomes were observed in some but not all root samples from the same plants. In samples taken from greenhouse-inoculated trees, levels of 'Ca. L. asiaticus' genomes varied systematically from 10(4) genomes/g at the graft inoculation site to 10(10) genomes/g in some leaf petioles. Root samples from these trees also contained 'Ca. L. asiaticus' at 10(7) genomes/g. In symptomatic fruit tissues, 'Ca. L. asiaticus' genomes were also readily detected and quantified. The highest

Antimicrobial effects of Citrus sinensis peel extracts against dental caries bacteria: An in vitro study

PubMed Central

Shetty, Sapna B.; Mahin-Syed-Ismail, Prabu; Varghese, Shaji; Thomas-George, Bibin; Kandathil- Thajuraj, Pathinettam; Baby, Deepak; Haleem, Shaista; Sreedhar, Sreeja

2016-01-01

Background Ethnomedicine is gaining admiration since years but still there is abundant medicinal flora which is unrevealed through research. The study was conducted to assess the in vitro antimicrobial potential and also determine the minimum inhibitory concentration (MIC) of Citrus sinensis peel extracts with a view of searching a novel extract as a remedy for dental caries pathogens. Material and Methods Aqueous and ethanol (cold and hot) extracts prepared from peel of Citrus sinensis were screened for in vitro antimicrobial activity against Streptococcus mutans and Lactobacillus acidophilus, using agar well diffusion method. The lowest concentration of every extract considered as the minimal inhibitory concentration (MIC) values were determined for both test organisms. One way ANOVA with Post Hoc Bonferroni test was applied for statistical analysis. Confidence level and level of significance were set at 95% and 5% respectively. Results Dental caries pathogens were inhibited most by hot ethanolic extract of Citrus sinensispeel followed by cold ethanolic extract. Aqueous extracts were effective at very high concentrations. Minimum inhibitory concentration of hot and cold ethanolic extracts of Citrus sinensis peel ranged between 12-15 mg/ml against both the dental caries pathogens. Conclusions Citrus sinensispeels extract was found to be effective against dental caries pathogens and contain compounds with therapeutic potential. Nevertheless, clinical trials on the effect of these plants are essential before advocating large-scale therapy. Key words:Agar well diffusion, antimicrobial activity, dental caries, Streptococcus mutans, Lactobacillus acidophilus. PMID:26855710

Inhibitory effects of crude extracts from several plants on postharvest pathogens of citrus

NASA Astrophysics Data System (ADS)

Gong, Mingfu; Guan, Qinlan; Xu, Shanshan

2018-04-01

China is one of the most important origin of citrus. Enormous economic losses was caused by fungal diseases in citrus harvest storage every year. The effective antimicrobial substances of garlic, ginger, celery and pepper were extracted by ethanol extraction and water extraction respectively. The inhibitory effects of the crude extract on Penicillium sp. caused fungal diseases in citrus harvest storage were also determined. The results showed that the extracts of garlic, ginger and celery had inhibitory effect on P. sp., but the extracts of pepper had no inhibitory effect on P. sp.. The garlic ethanol extracts had the best inhibitory effect on P. citrinum.

Delivery of gene biotechnologies to plants: Pathogen and pest control

USDA-ARS?s Scientific Manuscript database

Treatment of oligonucleotides to plants for host delivered suppression of microbes and insect pests of citrus was successful. FANA_ASO, (2'-deoxy-2'-fluoro-D- arabinonucleic acid)_( antisense oligonucleotides- AUM LifeTech) designed to: Asian citrus psyllid; Citrus plant bacterial pathogen of citru...

Huanglongbing: An overview of a complex pathosystem ravaging the world's citrus.

PubMed

da Graça, John V; Douhan, Greg W; Halbert, Susan E; Keremane, Manjunath L; Lee, Richard F; Vidalakis, Georgios; Zhao, Hongwei

2016-04-01

Citrus huanglongbing (HLB) has become a major disease and limiting factor of production in citrus areas that have become infected. The destruction to the affected citrus industries has resulted in a tremendous increase to support research that in return has resulted in significant information on both applied and basic knowledge concerning this important disease to the global citrus industry. Recent research indicates the relationship between citrus and the causal agent of HLB is shaped by multiple elements, in which host defense responses may also play an important role. This review is intended to provide an overview of the importance of HLB to a wider audience of plant biologists. Recent advances on host-pathogen interactions, population genetics and vectoring of the causal agent are discussed. © 2015 Institute of Botany, Chinese Academy of Sciences.

The Asian citrus psyllid host Murraya koenigii is immune to citrus Huanglongbing pathogen 'Candidatus Liberibacter asiaticus'.

PubMed

Beloti, Vitor Hugo; Alves, Gustavo; Coletta-Filho, Helvécio; Yamamoto, Pedro

2018-04-12

The Asian citrus psyllid (ACP) Diaphorina citri, vector of 'Candidatus Liberibacter asiaticus' (CLas), the putative causal agent of citrus Huanglongbing (HLB), is controlled by application of insecticides, which, although effective, has resulted in serious biological imbalances. New management tools are needed, and the technique known as 'trap crop' has been attracting attention. A potential plant for use as a trap crop in the management of the ACP is Murraya koenigii (curry leaf). However, for this plant to be used in the field, it needs to be attractive for the vector and must not harbor CLas. To verify the potential of curry leaf as trap crop for the management of HLB, we investigated the ability of D. citri to transmit CLas to M. koenigii, and to other test plants, including M. paniculata (orange jasmine) and 'Valencia' sweet-orange seedlings. For the tests, the insects were reared on a symptomatic CLas-infected plant and allowed to feed on the three test plant species. The overall maximum transmission rate for the citrus seedlings was 83.3%, and for orange jasmine was 33.3%. Successful transmission of CLas by ACP to the curry-leaf seedlings was not observed, and it was treated as immune to CLas. Supported by the previous results that M. koenigii is attractive for ACP, these results indicate that curry leaf is an excellent candidate for use as a trap crop, to improve the management of the insect vector and consequently of HLB.

Maculosin, a host-specific phytotoxin for spotted knapweed from Alternaria alternata

PubMed Central

Stierle, Andrea C.; Cardellina, John H.; Strobel, Gary A.

1988-01-01

Several diketopiperazines have been isolated from liquid cultures of Alternaria alternata, the causal agent of black leaf blight of spotted knapweed, Centaurea maculosa Lam. One of these compounds, maculosin [the diketopiperazine cyclo(-L-Pro-L-Tyr-)], was active in the nicked-leaf bioassay at 10-5 M; synthetic maculosin possessed chemical and biological activities identical to those of the natural product. Other diketopiperazines isolated from the fungus possessed either less activity or none at all. In tests against 19 plant species, maculosin was phytotoxic only to spotted knapweed. Thus maculosin is a host-specific phytotoxin from a weed pathogen. PMID:16593989

Amino acids implicated in plant defense are higher in Candidatus Liberibacter asiaticus-tolerant citrus varieties

PubMed Central

Killiny, Nabil; Hijaz, Faraj

2016-01-01

ABSTRACT Citrus Huanglongbing (HLB), also known as citrus greening, has been threatening the citrus industry since the early 1900's and up to this date there are no effective cures for this disease. Field observations and greenhouse controlled studies demonstrated that some citrus genotypes are more tolerant to Candidatus Liberibacter asiaticus (CLas) pathogen than others. However, the mechanisms underpinning tolerance has not been determined yet. The phloem sap composition of CLas-tolerant and sensitive citrus varieties was studied to identify metabolites that could be responsible for their tolerance to CLas. The citrus phloem sap was collected by centrifugation and was analyzed with gas chromatography-mass spectrometry after methyl chloroformate derivatization. Thirty-three metabolites were detected in the phloem sap of the studied varieties: twenty 20 amino acids, eight 8 organic acids, and five 5 fatty acids. Interestingly, the levels of most amino acids, especially those implicated in plantdefense to pathogens such as phenylalanine, tyrosine, tryptophan, lysine, and asparagine were higher in tolerant varieties. Although the level of organic acids varied between cultivars, this variation was not correlated with citrus resistance to CLas and could be cultivar specific. The fatty acids were found in trace amounts and in most cases their levels were not significantly different among varieties. Better understanding of the mechanisms underpinning citrus tolerance to CLas will help in developing economically tolerant varieties. PMID:27057814

Amino acids implicated in plant defense are higher in Candidatus Liberibacter asiaticus-tolerant citrus varieties.

PubMed

Killiny, Nabil; Hijaz, Faraj

2016-01-01

Citrus Huanglongbing (HLB), also known as citrus greening, has been threatening the citrus industry since the early 1900's and up to this date there are no effective cures for this disease. Field observations and greenhouse controlled studies demonstrated that some citrus genotypes are more tolerant to Candidatus Liberibacter asiaticus (CLas) pathogen than others. However, the mechanisms underpinning tolerance has not been determined yet. The phloem sap composition of CLas-tolerant and sensitive citrus varieties was studied to identify metabolites that could be responsible for their tolerance to CLas. The citrus phloem sap was collected by centrifugation and was analyzed with gas chromatography-mass spectrometry after methyl chloroformate derivatization. Thirty-three metabolites were detected in the phloem sap of the studied varieties: twenty 20 amino acids, eight 8 organic acids, and five 5 fatty acids. Interestingly, the levels of most amino acids, especially those implicated in plantdefense to pathogens such as phenylalanine, tyrosine, tryptophan, lysine, and asparagine were higher in tolerant varieties. Although the level of organic acids varied between cultivars, this variation was not correlated with citrus resistance to CLas and could be cultivar specific. The fatty acids were found in trace amounts and in most cases their levels were not significantly different among varieties. Better understanding of the mechanisms underpinning citrus tolerance to CLas will help in developing economically tolerant varieties.

Development of loop-mediated isothermal amplification and SYBR green real-time PCR methods for the detection of Citrus yellow mosaic badnavirus in citrus species.

PubMed

Anthony Johnson, A M; Dasgupta, I; Sai Gopal, D V R

2014-07-01

Citrus yellow mosaic badnavirus (CMBV) is an important pathogen in southern India spread by infected citrus propagules. One of the measures to arrest the spread of CMBV is to develop methods to screen and certify citrus propagules as CMBV-free. The methods loop-mediated isothermal amplification (LAMP) and SYBR green real-time PCR (SGRTPCR) have been developed for the efficient detection of CMBV in citrus propagules. This paper compares the sensitivities of LAMP and SGRTPCR with polymerase chain reaction (PCR) for the detection of CMBV. Whereas PCR and LAMP were able to detect CMBV from a minimum of 10 ng of total DNA of infected leaf samples, SGRTPCR could detect the same from 1 ng of total DNA. Using SGRTPCR, the viral titres were estimated to be the highest in rough lemon and lowest in Nagpur Mandarin of the five naturally infected citrus species tested. The results will help in designing suitable strategies for the sensitive detection of CMBV from citrus propagules. Copyright © 2014 Elsevier B.V. All rights reserved.

Characterization of a proposed dichorhavirus associated with the citrus leprosis disease and analysis of the host response.

PubMed

Cruz-Jaramillo, José Luis; Ruiz-Medrano, Roberto; Rojas-Morales, Lourdes; López-Buenfil, José Abel; Morales-Galván, Oscar; Chavarín-Palacio, Claudio; Ramírez-Pool, José Abrahán; Xoconostle-Cázares, Beatriz

2014-07-07

The causal agents of Citrus leprosis are viruses; however, extant diagnostic methods to identify them have failed to detect known viruses in orange, mandarin, lime and bitter orange trees with severe leprosis symptoms in Mexico, an important citrus producer. Using high throughput sequencing, a virus associated with citrus leprosis was identified, belonging to the proposed Dichorhavirus genus. The virus was termed Citrus Necrotic Spot Virus (CNSV) and contains two negative-strand RNA components; virions accumulate in the cytoplasm and are associated with plasmodesmata-channels interconnecting neighboring cells-suggesting a mode of spread within the plant. The present study provides insights into the nature of this pathogen and the corresponding plant response, which is likely similar to other pathogens that do not spread systemically in plants.

Characterization of a Proposed Dichorhavirus Associated with the Citrus Leprosis Disease and Analysis of the Host Response

PubMed Central

Cruz-Jaramillo, José Luis; Ruiz-Medrano, Roberto; Rojas-Morales, Lourdes; López-Buenfil, José Abel; Morales-Galván, Oscar; Chavarín-Palacio, Claudio; Ramírez-Pool, José Abrahán; Xoconostle-Cázares, Beatriz

2014-01-01

The causal agents of Citrus leprosis are viruses; however, extant diagnostic methods to identify them have failed to detect known viruses in orange, mandarin, lime and bitter orange trees with severe leprosis symptoms in Mexico, an important citrus producer. Using high throughput sequencing, a virus associated with citrus leprosis was identified, belonging to the proposed Dichorhavirus genus. The virus was termed Citrus Necrotic Spot Virus (CNSV) and contains two negative-strand RNA components; virions accumulate in the cytoplasm and are associated with plasmodesmata—channels interconnecting neighboring cells—suggesting a mode of spread within the plant. The present study provides insights into the nature of this pathogen and the corresponding plant response, which is likely similar to other pathogens that do not spread systemically in plants. PMID:25004279

Implementation of Citrus Shoot Tip Cryopreservation in the USDA-ARS National Plant Germplasm System

USDA-ARS?s Scientific Manuscript database

The USDA-ARS National Plant Germplasm System (NPGS) maintains 540 Citrus cultivars and crop wild relatives as duplicate clones in a screenhouse at the National Clonal Germplasm Repository for Citrus and Dates (NCGRCD) in Riverside, California. These 540 accessions are pathogen-tested and apparently ...

Characterization of Pseudomonas syringae pv. syringae, Causal Agent of Citrus Blast of Mandarin in Montenegro.

PubMed

Ivanović, Žarko; Perović, Tatjana; Popović, Tatjana; Blagojević, Jovana; Trkulja, Nenad; Hrnčić, Snježana

2017-02-01

Citrus blast caused by bacterium Pseudomonas syringae is a very important disease of citrus occuring in many areas of the world, but with few data about genetic structure of the pathogen involved. Considering the above fact, this study reports genetic characterization of 43 P. syringae isolates obtained from plant tissue displaying citrus blast symptoms on mandarin ( Citrus reticulata ) in Montenegro, using multilocus sequence analysis of gyrB , rpoD , and gap1 gene sequences. Gene sequences from a collection of 54 reference pathotype strains of P. syringae from the Plant Associated and Environmental Microbes Database (PAMDB) was used to establish a genetic relationship with our isolates obtained from mandarin. Phylogenetic analyses of gyrB , rpoD , and gap1 gene sequences showed that P. syringae pv. syringae causes citrus blast in mandarin in Montenegro, and belongs to genomospecies 1. Genetic homogeneity of isolates suggested that the Montenegrian population might be clonal which indicates a possible common source of infection. These findings may assist in further epidemiological studies of this pathogen and for determining mandarin breeding strategies for P. syringae control.

Characterization of Pseudomonas syringae pv. syringae, Causal Agent of Citrus Blast of Mandarin in Montenegro

PubMed Central

Ivanović, Žarko; Perović, Tatjana; Popović, Tatjana; Blagojević, Jovana; Trkulja, Nenad; Hrnčić, Snježana

2017-01-01

Citrus blast caused by bacterium Pseudomonas syringae is a very important disease of citrus occuring in many areas of the world, but with few data about genetic structure of the pathogen involved. Considering the above fact, this study reports genetic characterization of 43 P. syringae isolates obtained from plant tissue displaying citrus blast symptoms on mandarin (Citrus reticulata) in Montenegro, using multilocus sequence analysis of gyrB, rpoD, and gap1 gene sequences. Gene sequences from a collection of 54 reference pathotype strains of P. syringae from the Plant Associated and Environmental Microbes Database (PAMDB) was used to establish a genetic relationship with our isolates obtained from mandarin. Phylogenetic analyses of gyrB, rpoD, and gap1 gene sequences showed that P. syringae pv. syringae causes citrus blast in mandarin in Montenegro, and belongs to genomospecies 1. Genetic homogeneity of isolates suggested that the Montenegrian population might be clonal which indicates a possible common source of infection. These findings may assist in further epidemiological studies of this pathogen and for determining mandarin breeding strategies for P. syringae control. PMID:28167885

Possible role of plant volatiles in tolerance against huanglongbing in citrus

PubMed Central

Hijaz, Faraj; Nehela, Yasser; Killiny, Nabil

2016-01-01

abstract Volatile organic compounds (VOC) play an important role in protecting plants from insect and pathogen attack. In this study, we investigated the leaf volatile profiles of 14 citrus varieties. The VOC in citrus leaves were extracted with n-hexane and analyzed using gas chromatography-mass spectrometry (GC-MS). Overall, 4six volatile compounds were identified in the n-hexane extract from citrus leaves. Most of the detected compounds belonged to 3 main groups (monoterpenes, sesquiterpenes, and aliphatic aldehydes). Principle component analysis was used to examine the relative distribution of the studied varieties to each other. Interestingly, volatile profiles of varieties that are tolerant to Candidatus Liberibacter asiaticus (CLas) were different from those of the susceptible ones. Tolerant and moderately-tolerant cultivars contained relatively higher amounts of volatiles than susceptible varieties. In addition, tolerant varieties were also higher in specific compounds which are known for their antimicrobial activities. These compounds include Aldehydes (undecanal, neral, geranial, and citronellal) and some monoterpenes such as linalool, d-limonene, myrcene, α- and β- phellandrene. In addition, some sesquiterpene compounds including t-caryophellene, γ-elemene, β-elemene, germacrene D, and geranyl acetate were higher in tolerant and moderately tolerant cultivars. Severinia buxifolia which is known for its tolerance to CLas and many other pathogens contained higher levels of santalenes and coumarins. Our results indicated that citrus leaf volatiles might play a role in citrus tolerance to CLas. The results of this study may help in understanding of the mechanism of citrus tolerance against CLas. PMID:26829496

Possible role of plant volatiles in tolerance against huanglongbing in citrus.

PubMed

Hijaz, Faraj; Nehela, Yasser; Killiny, Nabil

2016-01-01

Volatile organic compounds (VOC) play an important role in protecting plants from insect and pathogen attack. In this study, we investigated the leaf volatile profiles of 14 citrus varieties. The VOC in citrus leaves were extracted with n-hexane and analyzed using gas chromatography-mass spectrometry (GC-MS). Overall, 4six volatile compounds were identified in the n-hexane extract from citrus leaves. Most of the detected compounds belonged to 3 main groups (monoterpenes, sesquiterpenes, and aliphatic aldehydes). Principle component analysis was used to examine the relative distribution of the studied varieties to each other. Interestingly, volatile profiles of varieties that are tolerant to Candidatus Liberibacter asiaticus (CLas) were different from those of the susceptible ones. Tolerant and moderately-tolerant cultivars contained relatively higher amounts of volatiles than susceptible varieties. In addition, tolerant varieties were also higher in specific compounds which are known for their antimicrobial activities. These compounds include Aldehydes (undecanal, neral, geranial, and citronellal) and some monoterpenes such as linalool, d-limonene, myrcene, α- and β- phellandrene. In addition, some sesquiterpene compounds including t-caryophellene, γ-elemene, β-elemene, germacrene D, and geranyl acetate were higher in tolerant and moderately tolerant cultivars. Severinia buxifolia which is known for its tolerance to CLas and many other pathogens contained higher levels of santalenes and coumarins. Our results indicated that citrus leaf volatiles might play a role in citrus tolerance to CLas. The results of this study may help in understanding of the mechanism of citrus tolerance against CLas.

Characterization of the Asian citrus psyllid transcriptome

USDA-ARS?s Scientific Manuscript database

The Asian citrus psyllid (Diaphorina citri Kuwayama) and other psyllids are important agricultural pests that cause extensive economic damage by feeding and as vectors of plant pathogens. No psyllid genomes have been characterized, and little is known about the composition of psyllid genomes or the ...

Presence of adhesive vesicles in the mycoherbicide Alternaria helianthi

USDA-ARS?s Scientific Manuscript database

Alternaria helianthi conidia have been shown to cause disease on common cocklebur. Conidia were applied to slides made hydrophobic by coating with dimethyldicholorosilane (mimics leaf surface), then rinsed and treated with FITC-Con A to stain the adhesive material. Alternaria helianthi coni...

Sesquiterpene emissions from Alternaria alternata and Fusarium oxysporum: Effects of age, nutrient availability, and co-cultivation

PubMed Central

Weikl, Fabian; Ghirardo, Andrea; Schnitzler, Jörg-Peter; Pritsch, Karin

2016-01-01

Alternaria alternata is one of the most studied fungi to date because of its impact on human life – from plant pathogenicity to allergenicity. However, its sesquiterpene emissions have not been systematically explored. Alternaria regularly co-occurs with Fusarium fungi, which are common plant pathogens, on withering plants. We analyzed the diversity and determined the absolute quantities of volatile organic compounds (VOCs) in the headspace above mycelial cultures of A. alternata and Fusarium oxysporum under different conditions (nutrient rich and poor, single cultures and co-cultivation) and at different mycelial ages. Using stir bar sorptive extraction and gas chromatography–mass spectrometry, we observed A. alternata to strongly emit sesquiterpenes, particularly during the early growth stages, while emissions from F. oxysporum consistently remained comparatively low. The emission profile characterizing A. alternata comprised over 20 sesquiterpenes with few effects from nutrient quality and age on the overall emission profile. Co-cultivation with F. oxysporum resulted in reduced amounts of VOCs emitted from A. alternata although its profile remained similar. Both fungi showed distinct emission profiles, rendering them suitable biomarkers for growth-detection of their phylotype in ambient air. The study highlights the importance of thorough and quantitative evaluations of fungal emissions of volatile infochemicals such as sesquiterpenes. PMID:26915756

Temporal occurrence and niche preferences of Phytophthora species causing brown rot of citrus in the Central Valley of California

USDA-ARS?s Scientific Manuscript database

Brown rot of citrus fruits is caused by several species of Phytophthora and is currently of serious concern for the California citrus industry. Two species, P. syringae and P. hibernalis, are quarantine pathogens in China, a major export market for California citrus. To maintain trade and estimate t...

[Expression of plant antimicrobial peptide pro-SmAMP2 gene increases resistance of transgenic potato plants to Alternaria and Fusarium pathogens].

PubMed

Vetchinkina, E M; Komakhina, V V; Vysotskii, D A; Zaitsev, D V; Smirnov, A N; Babakov, A V; Komakhin, R A

2016-09-01

The chickweed (Stellaria media L.) pro-SmAMP2 gene encodes the hevein-like peptides that have in vitro antimicrobial activity against certain harmful microorganisms. These peptides play an important role in protecting the chickweed plants from infection, and the pro-SmAMP2 gene was previously used to protect transgenic tobacco and Arabidopsis plants from phytopathogens. In this study, the pro-SmAMP2 gene under control of viral CaMV35S promoter or under control of its own pro-SmAMP2 promoter was transformed into cultivated potato plants of two cultivars, differing in the resistance to Alternaria: Yubiley Zhukova (resistant) and Skoroplodny (susceptible). With the help of quantitative real-time PCR, it was demonstrated that transgenic potato plants expressed the pro-SmAMP2 gene under control of both promoters at the level comparable to or exceeding the level of the potato actin gene. Assessment of the immune status of the transformants demonstrated that expression of antimicrobial peptide pro-SmAMP2 gene was able to increase the resistance to a complex of Alternaria sp. and Fusarium sp. phytopathogens only in potato plants of the Yubiley Zhukova cultivar. The possible role of the pro-SmAMP2 products in protecting potatoes from Alternaria sp. and Fusarium sp. is discussed.

Java project on periodontal diseases: effect of vitamin C/calcium threonate/citrus flavonoids supplementation on periodontal pathogens, CRP and HbA1c.

PubMed

Amaliya, Amaliya; Laine, Marja L; Loos, Bruno G; Van der Velden, Ubele

2015-12-01

To assess in a periodontally diseased rural population deprived from regular dental care and having poor dietary conditions, the effect of vitamin C/calcium threonate/citrus flavonoids (VitC/Ca/Fl) supplementation on subgingival microbiota and plasma levels of vitamin C, HbA1c and hsCRP. The study population consisted of 98 subjects who previously participated in a prospective study on the natural history of periodontitis. Participants were instructed to consume one tablet/day containing 200 mg Ester C(®) calcium ascorbate, 25 mg calcium threonate and 100 mg citrus flavonoids for 90 days. Following parameters were evaluated: prevalence/amount of seven traditional periodontal pathogens, cytomegalovirus, Epstein-Barr virus (EBV); and plasma levels of vitamin C, HbA1c and hsCRP. After VitC/Ca/Fl supplementation, 100% of subjects showed normal plasma vitamin C values compared to 55% before. At baseline, 48% of subjects harboured Aggregatibacter actinomycetemcomitans, >97% the other periodontal pathogens and 73% EBV. Supplementation with VitC/Ca/F reduced the subgingival load of all studied bacteria (p-values: 0.014-0.0001) and EBV (p < 0.0001) substantially in all initially positive subjects. Plasma levels of HbA1c and hsCRP dropped in all subjects (p < 0.0001). This uncontrolled study suggested that supplemental VitC/Ca/Fl may be helpful in reducing subgingival numbers of periodontal pathogens and EBV, and promoting systemic health. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Studies of Microbiota Dynamics Reveals Association of "Candidatus Liberibacter Asiaticus" Infection with Citrus (Citrus sinensis) Decline in South of Iran.

PubMed

Passera, Alessandro; Alizadeh, Hamidreza; Azadvar, Mehdi; Quaglino, Fabio; Alizadeh, Asma; Casati, Paola; Bianco, Piero A

2018-06-20

Citrus Decline Disease was recently reported to affect several citrus species in Iran when grafted on a local rootstock variety, Bakraee. Preliminary studies found “ Candidatus Phytoplasma aurantifoliae” and “ Candidatus Liberibacter asiaticus” as putative etiological agents, but were not ultimately able to determine which one, or if an association of both, were causing the disease. The current study has the aim of characterizing the microbiota of citrus plants that are either asymptomatic, showing early symptoms, or showing late symptoms through amplification of the V1⁻V3 region of 16S rRNA gene using an Illumina sequencer in order to (i) clarify the etiology of the disease, and (ii) describe the microbiota associated to different symptom stages. Our results suggest that liberibacter may be the main pathogen causing Citrus Decline Disease, but cannot rule out the possibility of phytoplasma being involved as well. The characterization of microbiota shows that the leaves show only two kinds of communities, either symptomatic or asymptomatic, while roots show clear distinction between early and late symptoms. These results could lead to the identification of bacteria that are related to successful plant defense response and, therefore, to immunity to the Citrus Decline Disease.

Absence of classical heat shock response in the citrus pathogen Xylella fastidiosa.

PubMed

Martins-de-Souza, Daniel; Martins, Daniel; Astua-Monge, Gustavo; Coletta-Filho, Helvécio Della; Winck, Flavia Vischi; Baldasso, Paulo Aparecido; de Oliveira, Bruno Menezes; Marangoni, Sérgio; Machado, Marcos Antônio; Novello, José Camillo; Smolka, Marcus Bustamante

2007-02-01

The fastidious bacterium Xylella fastidiosa is associated with important crop diseases worldwide. We have recently shown that X. fastidiosa is a peculiar organism having unusually low values of gene codon bias throughout its genome and, unexpectedly, in the group of the most abundant proteins. Here, we hypothesized that the lack of codon usage optimization in X. fastidiosa would incapacitate this organism to undergo quick and massive changes in protein expression as occurs in a classical stress response. Proteomic analysis of the response to heat stress in X. fastidiosa revealed that no changes in protein expression can be detected. Moreover, stress-inducible proteins identified in the closely related citrus pathogen Xanthomonas axonopodis pv citri were found to be constitutively expressed in X. fastidiosa. These proteins have extremely high codon bias values in the X. citri and other well-studied organisms, but low values in X. fastidiosa. Because biased codon usage is well known to correlate to the rate of protein synthesis, we speculate that the peculiar codon bias distribution in X. fastidiosa is related to the absence of a classical stress response, and, probably, alternative strategies for survival of X. fastidiosa under stressfull conditions.

Development of multiplex polymerase chain reaction assay for simultaneous detection of clostero-, badna- and mandari-viruses along with huanglongbing bacterium in citrus trees.

PubMed

Meena, Ram Prasnna; Baranwal, V K

2016-09-01

Citrus trees harbor a large number of viral and bacterial pathogens. Citrus yellow vein clearing virus (CYVCV), Indian citrus ringspot virus (ICRSV), Citrus yellow mosaic virus (CYMV), Citrus tristeza virus (CTV) and a bacterium, Candidatus Liberibacter asiaticus (CLa) associated with huanglongbing (HLB) disease, the most prevalent pathogens in citrus orchards of different regions in India and are responsible for debilitating citriculture. For detection of these viral and bacterial pathogens a quick, sensitive and cost effective detection method is required. With this objective a multiplex polymerase chain reaction (mPCR) assay was developed for simultaneous detection of four viruses and a bacterium in citrus. Several sets of primers were designed for each virus based on the retrieved reference sequences from the GenBank. A primer pair published previously was used for greening bacterium. Each pair of primers was evaluated for their sensitivity and differentiation by simplex and mPCR. The constant amplified products were identified on the basis of molecular size in mPCR and were compared with standard PCR. The amplicons were cloned and results were confirmed with sequencing analysis. The mPCR assay was validated using naturally infected field samples for one or more citrus viruses and the huanglongbing bacterium. The mPCR assay developed here will aid in the production of virus free planting materials and rapid indexing for certification of citrus budwood programme. Copyright © 2016 Elsevier B.V. All rights reserved.

Ultrastructure of the salivary glands, alimentary canal and bacteria-like organisms in the Asian citrus psyllid, vector of citrus huanglongbing-disease bacteria

USDA-ARS?s Scientific Manuscript database

Several psyllids (Hemiptera: Psylloidea) are known as vectors of some economically important viral and bacterial plant pathogens. The Asian citrus psyllid (ACP, Diaphorina citri, Hemiptera, Liviidae) is the principal vector of ‘Candidatus Liberibacter asiaticus’ (Las), the putative bacterial causal ...

The Distribution of Coumarins and Furanocoumarins in Citrus Species Closely Matches Citrus Phylogeny and Reflects the Organization of Biosynthetic Pathways

PubMed Central

Dugrand-Judek, Audray; Olry, Alexandre; Hehn, Alain; Costantino, Gilles; Ollitrault, Patrick; Froelicher, Yann; Bourgaud, Frédéric

2015-01-01

Citrus plants are able to produce defense compounds such as coumarins and furanocoumarins to cope with herbivorous insects and pathogens. In humans, these chemical compounds are strong photosensitizers and can interact with medications, leading to the “grapefruit juice effect”. Removing coumarins and furanocoumarins from food and cosmetics imply additional costs and might alter product quality. Thus, the selection of Citrus cultivars displaying low coumarin and furanocoumarin contents constitutes a valuable alternative. In this study, we performed ultra-performance liquid chromatography coupled with mass spectrometry analyses to determine the contents of these compounds within the peel and the pulp of 61 Citrus species representative of the genetic diversity all Citrus. Generally, Citrus peel contains larger diversity and higher concentrations of coumarin/furanocoumarin than the pulp of the same fruits. According to the chemotypes found in the peel, Citrus species can be separated into 4 groups that correspond to the 4 ancestral taxa (pummelos, mandarins, citrons and papedas) and extended with their respective secondary species descendants. Three of the 4 ancestral taxa (pummelos, citrons and papedas) synthesize high amounts of these compounds, whereas mandarins appear practically devoid of them. Additionally, all ancestral taxa and their hybrids are logically organized according to the coumarin and furanocoumarin pathways described in the literature. This organization allows hypotheses to be drawn regarding the biosynthetic origin of compounds for which the biogenesis remains unresolved. Determining coumarin and furanocoumarin contents is also helpful for hypothesizing the origin of Citrus species for which the phylogeny is presently not firmly established. Finally, this work also notes favorable hybridization schemes that will lead to low coumarin and furanocoumarin contents, and we propose to select mandarins and Ichang papeda as Citrus varieties for use in

Characterization of Citrus sinensis transcription factors closely associated with the non-host response to Xanthomonas campestris pv. vesicatoria.

PubMed

Daurelio, Lucas D; Romero, María S; Petrocelli, Silvana; Merelo, Paz; Cortadi, Adriana A; Talón, Manuel; Tadeo, Francisco R; Orellano, Elena G

2013-07-01

Plants, when exposed to certain pathogens, may display a form of genotype-independent resistance, known as non-host response. In this study, the response of Citrus sinensis (sweet orange) leaves to Xanthomonas campestris pv. vesicatoria (Xcv), a pepper and tomato pathogenic bacterium, was analyzed through biochemical assays and cDNA microarray hybridization and compared with Asiatic citrus canker infection caused by Xanthomonas citri subsp. citri. Citrus leaves exposed to the non-host bacterium Xcv showed hypersensitive response (HR) symptoms (cell death), a defense mechanism common in plants but poorly understood in citrus. The HR response was accompanied by differentially expressed genes that are associated with biotic stress and cell death. Moreover, 58 transcription factors (TFs) were differentially regulated by Xcv in citrus leaves, including 26 TFs from the stress-associated families AP2-EREBP, bZip, Myb and WRKY. Remarkably, in silico analysis of the distribution of expressed sequence tags revealed that 10 of the 58 TFs, belonging to C2C2-GATA, C2H2, CCAAT, HSF, NAC and WRKY gene families, were specifically over-represented in citrus stress cDNA libraries. This study identified candidate TF genes for the regulation of key steps during the citrus non-host HR. Furthermore, these TFs might be useful in future strategies of molecular breeding for citrus disease resistance. Copyright © 2013 Elsevier GmbH. All rights reserved.

Molecular Basis of Citrus sunki Susceptibility and Poncirus trifoliata Resistance Upon Phytophthora parasitica Attack.

PubMed

Dalio, Ronaldo José Durigan; Máximo, Heros José; Oliveira, Tiago Silva; Azevedo, Thamara de Medeiros; Felizatti, Henrique Leme; Campos, Magnólia de Araújo; Machado, Marcos Antonio

2018-03-01

Coevolution has shaped the molecular basis of an extensive number of defense mechanisms in plant-pathogen interactions. Phytophthora parasitica, a hemibiothrophic oomycete pathogen and the causal agent of citrus root rot and gummosis, interacts differently with Citrus sunki and Poncirus trifoliata, two commonly favored citrus rootstocks that are recognized as susceptible and resistant, respectively, to P. parasitica. The molecular core of these interactions remains elusive. Here, we provide evidence on the defense strategies employed by both susceptible and resistant citrus rootstocks, in parallel with P. parasitica deployment of effectors. Time course expression analysis (quantitative real-time polymerase chain reaction) of several defense-related genes were evaluated during i) plant disease development, ii) necrosis, and iii) pathogen effector gene expression. In C. sunki, P. parasitica deploys effectors, including elicitins, NPP1 (necrosis-inducing Phytophthora protein 1), CBEL (cellulose-binding elicitor and lectin activity), RxLR, and CRN (crinkler), and, consequently, this susceptible plant activates its main defense signaling pathways that result in the hypersensitive response and necrosis. Despite the strong plant-defense response, it fails to withstand P. parasitica invasion, confirming its hemibiothrophic lifestyle. In Poncirus trifoliata, the effectors were strongly expressed, nevertheless failing to induce any immunity manipulation and disease development, suggesting a nonhost resistance type, in which the plant relies on preformed biochemical and anatomical barriers.

Extracellular matrix-associated proteome changes during non-host resistance in citrus-Xanthomonas interactions.

PubMed

Swaroopa Rani, Tirupaati; Podile, Appa Rao

2014-04-01

Non-host resistance (NHR) is a most durable broad-spectrum resistance employed by the plants to restrict majority of pathogens. Plant extracellular matrix (ECM) is a critical defense barrier. Understanding ECM responses during interaction with non-host pathogen will provide insights into molecular events of NHR. In this study, the ECM-associated proteome was compared during interaction of citrus with pathogen Xanthomonas axonopodis pv. citri (Xac) and non-host pathogen Xanthomonas oryzae pv. oryzae (Xoo) at 8, 16, 24 and 48 h post inoculation. Comprehensive analysis of ECM-associated proteins was performed by extracting wall-bound and soluble ECM components using both destructive and non-destructive procedures. A total of 53 proteins was differentially expressed in citrus-Xanthomonas host and non-host interaction, out of which 44 were identified by mass spectrometry. The differentially expressed proteins were related to (1) defense-response (5 pathogenesis-related proteins, 3 miraculin-like proteins (MIR, MIR1 and MIR2) and 2 proteases); (2) enzymes of reactive oxygen species (ROS) metabolism [Cu/Zn superoxide dismutase (SOD), Fe-SOD, ascorbate peroxidase and 2-cysteine-peroxiredoxin]; (3) signaling (lectin, curculin-like lectin and concanavalin A-like lectin kinase); and (4) cell-wall modification (α-xylosidase, glucan 1, 3 β-glucosidase, xyloglucan endotransglucosylase/hydrolase). The decrease in ascorbate peroxidase and cysteine-peroxiredoxin could be involved in maintenance of ROS levels. Increase in defense, cell-wall remodeling and signaling proteins in citrus-Xoo interaction suggests an active involvement of ECM in execution of NHR. Partially compromised NHR in citrus against Xoo, upon Brefeldin A pre-treatment supported the role of non-classical secretory proteins in this phenomenon. © 2013 Scandinavian Plant Physiology Society.

Bostrycin and 4-deoxybostrycin: two nonspecific phytotoxins produced by Alternaria eichhorniae.

PubMed Central

Charudattan, R; Rao, K V

1982-01-01

Two crystalline red pigments with phytotoxic activity were isolated from culture filtrates of Alternaria eichhorniae, a pathogen of the water hyacinth Eichhornia crassipes. The pigments were present in the ratio of 4:1 and were identified as bostrycin and 4-deoxybostrycin, respectively. This is the first isolation of 4-deoxybostrycin from a natural source. Bostrycin, 4-deoxybostrycin, and their isopropylidene derivatives induced necrosis on tested plant leaves comparable to the A. eichhorniae-induced necrosis on water hyacinth. The lowest phytotoxic concentrations of crystalline bostrycin and 4-deoxybostrycin on water hyacinth leaves were about 7 and 30 microgram/ml, respectively. Both substances were inhibitory to Bacillus subtilis but were inactive against the fungus Geotrichum candidum. PMID:6805431

Bostrycin and 4-deoxybostrycin: two nonspecific phytotoxins produced by Alternaria eichhorniae.

PubMed

Charudattan, R; Rao, K V

1982-04-01

Two crystalline red pigments with phytotoxic activity were isolated from culture filtrates of Alternaria eichhorniae, a pathogen of the water hyacinth Eichhornia crassipes. The pigments were present in the ratio of 4:1 and were identified as bostrycin and 4-deoxybostrycin, respectively. This is the first isolation of 4-deoxybostrycin from a natural source. Bostrycin, 4-deoxybostrycin, and their isopropylidene derivatives induced necrosis on tested plant leaves comparable to the A. eichhorniae-induced necrosis on water hyacinth. The lowest phytotoxic concentrations of crystalline bostrycin and 4-deoxybostrycin on water hyacinth leaves were about 7 and 30 microgram/ml, respectively. Both substances were inhibitory to Bacillus subtilis but were inactive against the fungus Geotrichum candidum.

Oligo-DNA Custom Macroarray for Monitoring Major Pathogenic and Non-Pathogenic Fungi and Bacteria in the Phyllosphere of Apple Trees

PubMed Central

He, Ying-Hong; Isono, Sayaka; Shibuya, Makoto; Tsuji, Masaharu; Adkar Purushothama, Charith-Raj; Tanaka, Kazuaki; Sano, Teruo

2012-01-01

Background To monitor the richness in microbial inhabitants in the phyllosphere of apple trees cultivated under various cultural and environmental conditions, we developed an oligo-DNA macroarray for major pathogenic and non-pathogenic fungi and bacteria inhabiting the phyllosphere of apple trees. Methods and Findings First, we isolated culturable fungi and bacteria from apple orchards by an agar-plate culture method, and detected 32 fungal and 34 bacterial species. Alternaria, Aureobasidium, Cladosporium, Rhodotorula, Cystofilobasidium, and Epicoccum genera were predominant among the fungi, and Bacillus, Pseudomonas, Sphingomonas, Methylobacterium, and Pantoea genera were predominant among the bacteria. Based on the data, we selected 29 major non-pathogenic and 12 phytopathogenic fungi and bacteria as the targets of macroarray. Forty-one species-specific 40-base pair long oligo-DNA sequences were selected from the nucleotide sequences of rDNA-internal transcribed spacer region for fungi and 16S rDNA for bacteria. The oligo-DNAs were fixed on nylon membrane and hybridized with digoxigenin-labeled cRNA probes prepared for each species. All arrays except those for Alternaria, Bacillus, and their related species, were specifically hybridized. The array was sensitive enough to detect 103 CFU for Aureobasidium pullulans and Bacillus cereus. Nucleotide sequencing of 100 each of independent fungal rDNA-ITS and bacterial 16S-rDNA sequences from apple tree was in agreement with the macroarray data obtained using the same sample. Finally, we analyzed the richness in the microbial inhabitants in the samples collected from apple trees in four orchards. Major apple pathogens that cause scab, Alternaria blotch, and Marssonina blotch were detected along with several non-phytopathogenic fungal and bacterial inhabitants. Conclusions The macroarray technique presented here is a strong tool to monitor the major microbial species and the community structures in the phyllosphere of

Oligo-DNA custom macroarray for monitoring major pathogenic and non-pathogenic fungi and bacteria in the phyllosphere of apple trees.

PubMed

He, Ying-Hong; Isono, Sayaka; Shibuya, Makoto; Tsuji, Masaharu; Adkar Purushothama, Charith-Raj; Tanaka, Kazuaki; Sano, Teruo

2012-01-01

To monitor the richness in microbial inhabitants in the phyllosphere of apple trees cultivated under various cultural and environmental conditions, we developed an oligo-DNA macroarray for major pathogenic and non-pathogenic fungi and bacteria inhabiting the phyllosphere of apple trees. First, we isolated culturable fungi and bacteria from apple orchards by an agar-plate culture method, and detected 32 fungal and 34 bacterial species. Alternaria, Aureobasidium, Cladosporium, Rhodotorula, Cystofilobasidium, and Epicoccum genera were predominant among the fungi, and Bacillus, Pseudomonas, Sphingomonas, Methylobacterium, and Pantoea genera were predominant among the bacteria. Based on the data, we selected 29 major non-pathogenic and 12 phytopathogenic fungi and bacteria as the targets of macroarray. Forty-one species-specific 40-base pair long oligo-DNA sequences were selected from the nucleotide sequences of rDNA-internal transcribed spacer region for fungi and 16S rDNA for bacteria. The oligo-DNAs were fixed on nylon membrane and hybridized with digoxigenin-labeled cRNA probes prepared for each species. All arrays except those for Alternaria, Bacillus, and their related species, were specifically hybridized. The array was sensitive enough to detect 10(3) CFU for Aureobasidium pullulans and Bacillus cereus. Nucleotide sequencing of 100 each of independent fungal rDNA-ITS and bacterial 16S-rDNA sequences from apple tree was in agreement with the macroarray data obtained using the same sample. Finally, we analyzed the richness in the microbial inhabitants in the samples collected from apple trees in four orchards. Major apple pathogens that cause scab, Alternaria blotch, and Marssonina blotch were detected along with several non-phytopathogenic fungal and bacterial inhabitants. The macroarray technique presented here is a strong tool to monitor the major microbial species and the community structures in the phyllosphere of apple trees and identify key species

Partial Resistance of Carrot to Alternaria dauci Correlates with In Vitro Cultured Carrot Cell Resistance to Fungal Exudates

PubMed Central

Voisine, Linda; Gatto, Julia; Hélesbeux, Jean-Jacques; Séraphin, Denis; Peña-Rodriguez, Luis M.; Richomme, Pascal; Boedo, Cora; Yovanopoulos, Claire; Gyomlai, Melvina; Briard, Mathilde; Simoneau, Philippe; Poupard, Pascal; Berruyer, Romain

2014-01-01

Although different mechanisms have been proposed in the recent years, plant pathogen partial resistance is still poorly understood. Components of the chemical warfare, including the production of plant defense compounds and plant resistance to pathogen-produced toxins, are likely to play a role. Toxins are indeed recognized as important determinants of pathogenicity in necrotrophic fungi. Partial resistance based on quantitative resistance loci and linked to a pathogen-produced toxin has never been fully described. We tested this hypothesis using the Alternaria dauci – carrot pathosystem. Alternaria dauci, causing carrot leaf blight, is a necrotrophic fungus known to produce zinniol, a compound described as a non-host selective toxin. Embryogenic cellular cultures from carrot genotypes varying in resistance against A. dauci were confronted with zinniol at different concentrations or to fungal exudates (raw, organic or aqueous extracts). The plant response was analyzed through the measurement of cytoplasmic esterase activity, as a marker of cell viability, and the differentiation of somatic embryos in cellular cultures. A differential response to toxicity was demonstrated between susceptible and partially resistant genotypes, with a good correlation noted between the resistance to the fungus at the whole plant level and resistance at the cellular level to fungal exudates from raw and organic extracts. No toxic reaction of embryogenic cultures was observed after treatment with the aqueous extract or zinniol used at physiological concentration. Moreover, we did not detect zinniol in toxic fungal extracts by UHPLC analysis. These results suggest that strong phytotoxic compounds are present in the organic extract and remain to be characterized. Our results clearly show that carrot tolerance to A. dauci toxins is one component of its partial resistance. PMID:24983469

Tale of the Huanglongbing Disease Pyramid in the Context of the Citrus Microbiome.

PubMed

Wang, Nian; Stelinski, Lukasz L; Pelz-Stelinski, Kirsten S; Graham, James H; Zhang, Yunzeng

2017-04-01

The Huanglongbing (HLB) disease pyramid is composed of Liberibacters, psyllid vectors, citrus hosts, and the environment. The epidemiological outcomes for Liberibacter-associated plant diseases are collectively determined by the inherent relationships among plant-Liberibacters-psyllids, and how various environmental factors affect plant-Liberibacter-psyllid interactions. Citrus-Liberibacter-psyllid interactions occur in a complex microbiome system. In this review, we focus on the progress in understanding the HLB disease pyramid, and how the microbiome affects the HLB disease pyramid including the interaction between HLB and the citrus microbiome; the interaction between Liberibacters and psyllids; the interaction between Liberibacters and gut microbiota in psyllids; and the effect of HLB on selected above- and belowground citrus pathogens. Their implications for HLB management are also discussed.

Detection of Alternaria fungal contamination in cereal grains by a polymerase chain reaction-based assay.

PubMed

Zur, Gideon; Shimoni, Eyal; Hallerman, Eric; Kashi, Yechezkel

2002-09-01

Alternaria sp. are important fungal contaminants of grain products; they secrete four structural classes of compounds that are toxic or carcinogenic to plants and animals and cause considerable economic losses to growers and the food-processing industry. Alternaria toxins have been detected by high-performance liquid chromatography (HPLC), enzyme-linked immunosorbent assay, and other techniques. Here, we report the development of a polymerase chain reaction (PCR)-based method for the detection of Alternaria DNA. PCR primers were designed to anneal to the ITS1 and ITS2 regions of the 5.8S rDNA gene of Alternaria alternata or Alternaria solani but not to other microbial or plant DNA. We compared the sensitivity of PCR in detecting Alternaria DNA, that of the HPLC method in detecting Alternaria alternariol and alternariol methyl ether toxins, and that of the morphological examination of mycelia and conidia in experimentally infested corn samples. The sensitivity of toxin detection for HPLC was above the level of contamination in a set of commercially obtained grain samples, resulting in negative scores for all samples, while the PCR-based method and mold growth plating followed by morphological identification of Alternaria gave parallel, positive results for 8 of 10 samples. The PCR assay required just 8 h, enabling the rapid and simultaneous testing of many samples at a low cost. PCR-based evidence for the presence of Alternaria DNA followed by positive assay results for Alternaria toxins would support the rejection of a shipment of grain.

Deciphering the bacterial microbiome of citrus plants in response to ‘Candidatus Liberibacter asiaticus’-infection and antibiotic treatment

USDA-ARS?s Scientific Manuscript database

Huanglongbing (HLB), the most devastating citrus disease worldwide, is vectored by phloem-feeding insects, and the pathogen in the USA is Candidatus Liberibacter asiaticus (Las). The bacterial microbiome of citrus after Las-infection and treatments with ampicillin (Amp) and gentamicin (Gm) was chara...

LRR-RLK family from two Citrus species: genome-wide identification and evolutionary aspects.

PubMed

Magalhães, Diogo M; Scholte, Larissa L S; Silva, Nicholas V; Oliveira, Guilherme C; Zipfel, Cyril; Takita, Marco A; De Souza, Alessandra A

2016-08-12

Leucine-rich repeat receptor-like kinases (LRR-RLKs) represent the largest subfamily of plant RLKs. The functions of most LRR-RLKs have remained undiscovered, and a few that have been experimentally characterized have been shown to have important roles in growth and development as well as in defense responses. Although RLK subfamilies have been previously studied in many plants, no comprehensive study has been performed on this gene family in Citrus species, which have high economic importance and are frequent targets for emerging pathogens. In this study, we performed in silico analysis to identify and classify LRR-RLK homologues in the predicted proteomes of Citrus clementina (clementine) and Citrus sinensis (sweet orange). In addition, we used large-scale phylogenetic approaches to elucidate the evolutionary relationships of the LRR-RLKs and further narrowed the analysis to the LRR-XII group, which contains several previously described cell surface immune receptors. We built integrative protein signature databases for Citrus clementina and Citrus sinensis using all predicted protein sequences obtained from whole genomes. A total of 300 and 297 proteins were identified as LRR-RLKs in C. clementina and C. sinensis, respectively. Maximum-likelihood phylogenetic trees were estimated using Arabidopsis LRR-RLK as a template and they allowed us to classify Citrus LRR-RLKs into 16 groups. The LRR-XII group showed a remarkable expansion, containing approximately 150 paralogs encoded in each Citrus genome. Phylogenetic analysis also demonstrated the existence of two distinct LRR-XII clades, each one constituted mainly by RD and non-RD kinases. We identified 68 orthologous pairs from the C. clementina and C. sinensis LRR-XII genes. In addition, among the paralogs, we identified a subset of 78 and 62 clustered genes probably derived from tandem duplication events in the genomes of C. clementina and C. sinensis, respectively. This work provided the first comprehensive

Amplification of DNA of Xanthomonas axonopodis pv. citri from historic citrus canker herbarium specimens.

PubMed

Li, Wenbin; Brlansky, Ronald H; Hartung, John S

2006-05-01

Herbaria are important resources for the study of the origins and dispersal of plant pathogens, particularly bacterial plant pathogens that incite local lesions in which large numbers of pathogen genomes are concentrated. Xanthomonas axonopodis pv. citri (Xac), the causal agent of citrus bacterial canker disease, is a notable example of such a pathogen. The appearance of novel strains of the pathogen in Florida and elsewhere make it increasingly important to understand the relationships among strains of this pathogen. USDA-ARS at Beltsville, Maryland maintains approximately 700 herbarium specimens with citrus canker disease lesions up to 90 years old, originally collected from all over the world, and so is an important resource for phytogeographic studies of this bacterium. Unfortunately, DNA in herbarium specimens is degraded and may contain high levels of inhibitors of PCR. In this study, we compared a total of 23 DNA isolation techniques in combination with 31 novel primer pairs in order to develop an efficient protocol for the analysis of Xac DNA in herbarium specimens. We identified the most reliable extraction method, identified in terms of successful amplification by our panel of 31 primer pairs. We also identified the most robust primer pairs, identified as successful in the largest number of extracts prepared by different methods. We amplified Xac genomic sequences up to 542 bp long from herbarium samples up to 89 years old. Primers varied in effectiveness, with some primer pairs amplifying Xac DNA from a 1/10,000 dilution of extract from a single lesion from a citrus canker herbarium specimen. Our methodology will be useful to identify pathogens and perform molecular analyses of bacterial and possibly fungal genomes from herbarium specimens.

[Determination of the profiles of secondary metabolites characteristic of Alternaria strains isolated from tomato].

PubMed

Benavidez Rozo, Martha Elizabeth; Patriarca, Andrea; Cabrera, Gabriela; Fernández Pinto, Virginia E

2014-01-01

Many Alternaria species have been studied for their ability to produce bioactive secondary metabolites, such as tentoxin (TEN), some of which have toxic properties. The main food contaminant toxins are tenuazonic acid, alternariol (AOH), alternariol monomethyl ether (AME), altenuene, and altertoxins i, ii and iii. To determine the profiles of secondary metabolites characteristic of Alternaria strains isolated from tomato for their chemotaxonomic classification. The profiles of secondary metabolites were determined by HPLC MS. The Alternaria isolates obtained from spoiled tomatoes belong, according to their morphological characteristics, to the species groups Alternaria alternata, Alternaria tenuissima and Alternaria arborescens, with A. tenuissima being the most frequent. The most frequent profiles of secondary metabolites belonging to the species groups A. alternata (AOH, AME, TEN), A. tenuissima (AOH, AME, TEN, tenuazonic acid) and A. arborescens (AOH, AME, TEN, tenuazonic acid) were determined, with some isolates of the latter being able to synthesize AAL toxins. Secondary metabolite profiles are a useful tool for the differentiation of small spored Alternaria isolates not easily identifiable by their morphological characteristics. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

First Report of Leaf Spot Caused by Alternaria tenuissima on Black Chokeberry (Aronia melanocarpa) in Korea.

PubMed

Wee, Jung-In; Park, Jong-Han; Back, Chang-Gi; You, Young-Hyun; Chang, Taehyun

2016-09-01

In July 2015, diseased leaves of black chokeberry ( Aronia melanocarpa ) were observed in Danyang and Gochang, Korea. The symptoms appeared as circular or irregular brown leaf spots, from which Alternaria tenuissima was isolated. The isolates were cultured on potato dextrose agar, and their morphological characteristics were observed under a light microscope. The colonies were whitish to ash colored. The pathogenicity test on healthy black chokeberry leaves produced circular brown spots, in line with the original symptoms. Molecular analyses of the ITS, GPD, RPB2, and TEF genes were conducted to confirm the identity of the pathogen. The phylogeny of the multi-gene sequences indicated that the causal agent was A. tenuissima . This study is the first report of A. tenuissima leaf spot on black chokeberry ( A. melanocarpa ).

Differential gene expression in Alternaria gaisen exposed to dark and light

USDA-ARS?s Scientific Manuscript database

Character states observed during sporulation have been the basis for segregation and description of many of the small-spored species of Alternaria. Phylogenetic analyses of ITS and housekeeping genes from small-spored Alternaria spp. do not support most of the currently defined morphological specie...

Xylella genomics and bacterial pathogenicity to plants.

PubMed

Dow, J M; Daniels, M J

2000-12-01

Xylella fastidiosa, a pathogen of citrus, is the first plant pathogenic bacterium for which the complete genome sequence has been published. Inspection of the sequence reveals high relatedness to many genes of other pathogens, notably Xanthomonas campestris. Based on this, we suggest that Xylella possesses certain easily testable properties that contribute to pathogenicity. We also present some general considerations for deriving information on pathogenicity from bacterial genomics. Copyright 2000 John Wiley & Sons, Ltd.

Local genetic diversity of Xanthomonas citri subsp. citri in citrus orchards in northwest Paraná state, Brazil

USDA-ARS?s Scientific Manuscript database

Xanthomonas citri subsp. citri, causal agent of Asiatic citrus canker, is an important pathogen of citrus in Brazil and elsewhere. The genetic diversity of X. citri subsp. citri pathtype ‘A’ has not been studied in Brazil at a local scale (up to 300 km). A total of 40 isolates were collected from le...

Characteristics of the perception of different severity measures of citrus canker and the relations between the various symptom types

USDA-ARS?s Scientific Manuscript database

Citrus canker is a disease of citrus and is caused by the bacterial pathogen Xanthomonas axonopodis pv citri (Xac). Ways of managing the disease are being sought, and accurate, precise, reproducible disease assessment is needed for monitoring epidemics. The objective of this study was to investigate...

The Evolutionary History and Spatiotemporal Dynamics of the NC Lineage of Citrus Tristeza Virus.

PubMed

Benítez-Galeano, María José; Castells, Matías; Colina, Rodney

2017-10-12

Citrus tristeza virus (CTV) is a major pathogen affecting citrus trees worldwide. However, few studies have focused on CTV's evolutionary history and geographic behavior. CTV is locally dispersed by an aphid vector and long distance dispersion due to transportation of contaminated material. With the aim to delve deeper into the CTV-NC (New Clade) genotype evolution, we estimated an evolution rate of 1.19 × 10 -3 subs/site/year and the most common recent ancestor in 1977. Furthermore, the place of origin of the genotype was in the United States, and a great expansion of the population was observed in Uruguay. This expansion phase could be a consequence of the increment in the number of naïve citrus trees in Uruguayan orchards encompassing citrus industry growth in the past years.

The quest for a non-vector psyllid: Natural variation in acquisition and transmission of the huanglongbing pathogen ‘Candidatus Liberibacter asiaticus’ by Asian citrus psyllid isofemale lines

PubMed Central

Ammar, El-Desouky; Hall, David G.; Hosseinzadeh, Saeed

2018-01-01

Genetic variability in insect vectors is valuable to study vector competence determinants and to select non-vector populations that may help reduce the spread of vector-borne pathogens. We collected and tested vector competency of 15 isofemale lines of Asian citrus psyllid, Diaphorina citri, vector of ‘Candidatus Liberibacter asiaticus’ (CLas). CLas is associated with huanglongbing (citrus greening), the most serious citrus disease worldwide. D. citri adults were collected from orange jasmine (Murraya paniculata) hedges in Florida, and individual pairs (females and males) were caged on healthy Murraya plants for egg laying. The progeny from each pair that tested CLas-negative by qPCR were maintained on Murraya plants and considered an isofemale line. Six acquisition tests on D. citri adults that were reared as nymphs on CLas-infected citrus, from various generations of each line, were conducted to assess their acquisition rates (percentage of qPCR-positive adults). Three lines with mean acquisition rates of 28 to 32%, were classified as ‘good’ acquirers and three other lines were classified as ‘poor’ acquirers, with only 5 to 8% acquisition rates. All lines were further tested for their ability to inoculate CLas by confining CLas-exposed psyllids for one week onto healthy citrus leaves (6–10 adults/leaf/week), and testing the leaves for CLas by qPCR. Mean inoculation rates were 19 to 28% for the three good acquirer lines and 0 to 3% for the three poor acquirer lines. Statistical analyses indicated positive correlations between CLas acquisition and inoculation rates, as well as between CLas titer in the psyllids and CLas acquisition or inoculation rates. Phenotypic and molecular characterization of one of the good and one of the poor acquirer lines revealed differences between them in color morphs and hemocyanin expression, but not the composition of bacterial endosymbionts. Understanding the genetic architecture of CLas transmission will enable the

First Report of Leaf Spot Caused by Alternaria tenuissima on Black Chokeberry (Aronia melanocarpa) in Korea

PubMed Central

Wee, Jung-In; Park, Jong-Han; Back, Chang-Gi; You, Young-Hyun

2016-01-01

In July 2015, diseased leaves of black chokeberry (Aronia melanocarpa) were observed in Danyang and Gochang, Korea. The symptoms appeared as circular or irregular brown leaf spots, from which Alternaria tenuissima was isolated. The isolates were cultured on potato dextrose agar, and their morphological characteristics were observed under a light microscope. The colonies were whitish to ash colored. The pathogenicity test on healthy black chokeberry leaves produced circular brown spots, in line with the original symptoms. Molecular analyses of the ITS, GPD, RPB2, and TEF genes were conducted to confirm the identity of the pathogen. The phylogeny of the multi-gene sequences indicated that the causal agent was A. tenuissima. This study is the first report of A. tenuissima leaf spot on black chokeberry (A. melanocarpa). PMID:27790070

Development and systematic validation of qPCR assays for rapid and reliable differentiation of Xylella fastidiosa strains causing citrus variegated chlorosis.

PubMed

Li, Wenbin; Teixeira, Diva C; Hartung, John S; Huang, Qi; Duan, Yongping; Zhou, Lijuan; Chen, Jianchi; Lin, Hong; Lopes, Silvio; Ayres, A Juliano; Levy, Laurene

2013-01-01

The xylem-limited, Gram-negative, fastidious plant bacterium Xylella fastidiosa is the causal agent of citrus variegated chlorosis (CVC), a destructive disease affecting approximately half of the citrus plantations in the State of São Paulo, Brazil. The disease was recently found in Central America and is threatening the multi-billion U.S. citrus industry. Many strains of X. fastidiosa are pathogens or endophytes in various plants growing in the U.S., and some strains cross infect several host plants. In this study, a TaqMan-based assay targeting the 16S rDNA signature region was developed for the identification of X. fastidiosa at the species level. Another TaqMan-based assay was developed for the specific identification of the CVC strains. Both new assays have been systematically validated in comparison with the primer/probe sets from four previously published assays on one platform and under similar PCR conditions, and shown to be superior. The species specific assay detected all X. fastidiosa strains and did not amplify any other citrus pathogen or endophyte tested. The CVC-specific assay detected all CVC strains but did not amplify any non-CVC X. fastidiosa nor any other citrus pathogen or endophyte evaluated. Both sets were multiplexed with a reliable internal control assay targeting host plant DNA, and their diagnostic specificity and sensitivity remained unchanged. This internal control provides quality assurance for DNA extraction, performance of PCR reagents, platforms and operators. The limit of detection for both assays was equivalent to 2 to 10 cells of X. fastidiosa per reaction for field citrus samples. Petioles and midribs of symptomatic leaves of sweet orange harbored the highest populations of X. fastidiosa, providing the best materials for detection of the pathogen. These new species specific assay will be invaluable for molecular identification of X. fastidiosa at the species level, and the CVC specific assay will be very powerful for the

Description of a novel mild strain of Citrus tristeza virus in California that reacts with monoclonal antibody MCA13

USDA-ARS?s Scientific Manuscript database

Quick decline caused by Citrus tristeza virus (CTV) destroyed citrus on sour orange rootstock in southern California in the 1930’s -40’s. However, use of resistant/tolerant CTV rootstocks, certified pathogen-free budwood, and quarantines have limited further economic damage from CTV. Multi-locus mar...

7 CFR 301.76-3 - Quarantined areas; citrus greening and Asian citrus psyllid.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 5 2013-01-01 2013-01-01 false Quarantined areas; citrus greening and Asian citrus... Greening and Asian Citrus Psyllid § 301.76-3 Quarantined areas; citrus greening and Asian citrus psyllid... quarantined area for Asian citrus psyllid in accordance with the criteria listed in paragraph (c) of this...

7 CFR 301.76-3 - Quarantined areas; citrus greening and Asian citrus psyllid.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 5 2011-01-01 2011-01-01 false Quarantined areas; citrus greening and Asian citrus... Greening and Asian Citrus Psyllid § 301.76-3 Quarantined areas; citrus greening and Asian citrus psyllid... quarantined area for Asian citrus psyllid in accordance with the criteria listed in paragraph (c) of this...

7 CFR 301.76-3 - Quarantined areas; citrus greening and Asian citrus psyllid.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 5 2012-01-01 2012-01-01 false Quarantined areas; citrus greening and Asian citrus... Greening and Asian Citrus Psyllid § 301.76-3 Quarantined areas; citrus greening and Asian citrus psyllid... quarantined area for Asian citrus psyllid in accordance with the criteria listed in paragraph (c) of this...

7 CFR 301.76-3 - Quarantined areas; citrus greening and Asian citrus psyllid.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 5 2014-01-01 2014-01-01 false Quarantined areas; citrus greening and Asian citrus... Greening and Asian Citrus Psyllid § 301.76-3 Quarantined areas; citrus greening and Asian citrus psyllid... quarantined area for Asian citrus psyllid in accordance with the criteria listed in paragraph (c) of this...

Citrus Genomics

PubMed Central

Talon, Manuel; Gmitter Jr., Fred G.

2008-01-01

Citrus is one of the most widespread fruit crops globally, with great economic and health value. It is among the most difficult plants to improve through traditional breeding approaches. Currently, there is risk of devastation by diseases threatening to limit production and future availability to the human population. As technologies rapidly advance in genomic science, they are quickly adapted to address the biological challenges of the citrus plant system and the world's industries. The historical developments of linkage mapping, markers and breeding, EST projects, physical mapping, an international citrus genome sequencing project, and critical functional analysis are described. Despite the challenges of working with citrus, there has been substantial progress. Citrus researchers engaged in international collaborations provide optimism about future productivity and contributions to the benefit of citrus industries worldwide and to the human population who can rely on future widespread availability of this health-promoting and aesthetically pleasing fruit crop. PMID:18509486

The SLT2 mitogen-activated protein kinase-mediated signalling pathway governs conidiation, morphogenesis, fungal virulence and production of toxin and melanin in the tangerine pathotype of Alternaria alternata.

PubMed

Yago, Jonar Ingan; Lin, Ching-Hsuan; Chung, Kuang-Ren

2011-09-01

Fungi respond and adapt to different environmental stimuli via signal transduction systems. We determined the function of a yeast SLT2 mitogen-activated protein (MAP) kinase homologue (AaSLT2) in Alternaria alternata, the fungal pathogen of citrus. Analysis of the loss-of-function mutant indicated that AaSLT2 is required for the production of a host-selective toxin, and is crucial for fungal pathogenicity. Moreover, the A. alternata slt2 mutants displayed hypersensitivity to cell wall-degrading enzymes and chemicals such as Calcofluor white and Congo red. This implicates an important role of AaSLT2 in the maintenance of cell wall integrity in A. alternata. The A. alternata slt2 mutants were also hypersensitive to a heteroaromatic compound, 2-chloro-5-hydroxypyridine, and a plant growth regulator, 2,3,5-triiodobenzoic acid. Developmentally, the AaSLT2 gene product was shown to be critical for conidial formation and hyphal elongation. Compared with the wild-type, the mutants produced fewer but slightly larger conidia with less transverse septae. The mutants also accumulated lower levels of melanin and chitin. Unlike the wild-type progenitor, the A. alternata slt2 mutants produced globose, swollen hyphae that did not elongate in a straight radial direction. All defective phenotypes in the mutant were restored by transformation and expression of a wild-type copy of AaSLT2 under the control of its endogenous promoter. This study highlights an important role of the AaSLT2 MAP kinase-mediated signalling pathway, regulating diverse physiological, developmental and pathological functions, in the tangerine pathotype of A. alternata. © 2011 THE AUTHORS. MOLECULAR PLANT PATHOLOGY © 2011 BSPP AND BLACKWELL PUBLISHING LTD.

7 CFR 301.76-2 - Regulated articles for Asian citrus psyllid and citrus greening.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 5 2011-01-01 2011-01-01 false Regulated articles for Asian citrus psyllid and citrus... Greening and Asian Citrus Psyllid § 301.76-2 Regulated articles for Asian citrus psyllid and citrus greening. The following are regulated articles for Asian citrus psyllid and citrus greening: (a) All plants...

7 CFR 301.76-2 - Regulated articles for Asian citrus psyllid and citrus greening.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 5 2014-01-01 2014-01-01 false Regulated articles for Asian citrus psyllid and citrus... Greening and Asian Citrus Psyllid § 301.76-2 Regulated articles for Asian citrus psyllid and citrus greening. The following are regulated articles for Asian citrus psyllid and citrus greening: (a) All plants...

7 CFR 301.76-2 - Regulated articles for Asian citrus psyllid and citrus greening.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 5 2012-01-01 2012-01-01 false Regulated articles for Asian citrus psyllid and citrus... Greening and Asian Citrus Psyllid § 301.76-2 Regulated articles for Asian citrus psyllid and citrus greening. The following are regulated articles for Asian citrus psyllid and citrus greening: (a) All plants...

7 CFR 301.76-2 - Regulated articles for Asian citrus psyllid and citrus greening.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 5 2013-01-01 2013-01-01 false Regulated articles for Asian citrus psyllid and citrus... Greening and Asian Citrus Psyllid § 301.76-2 Regulated articles for Asian citrus psyllid and citrus greening. The following are regulated articles for Asian citrus psyllid and citrus greening: (a) All plants...

Localization and dynamics of Wolbachia infection in Asian citrus psyllid Diaphorina citri, the insect vector of the causal pathogens of Huanglongbing.

PubMed

Ren, Su-Li; Li, Yi-Han; Ou, Da; Guo, Yan-Jun; Qureshi, Jawwad A; Stansly, Philip A; Qiu, Bao-Li

2018-03-23

Wolbachia is a group of intracellular bacteria that infect a wide range of arthropods including the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama. This insect is the vector of Candidatus Liberibacter asiaticus (CLas), the causal pathogen of Huanglongbing or citrus greening disease. Here, we investigated the localization pattern and infection dynamics of Wolbachia in different developmental stages of ACP. Results revealed that all developmental stages of ACP including egg, 1st-5th instar nymphs, and adults of both gender were infected with Wolbachia. FISH visualization of an ACP egg showed that Wolbachia moved from the egg stalk of newly laid eggs to a randomly distributed pattern throughout the egg prior to hatching. The infection rate varied between nymphal instars. The titers of Wolbachia in fourth and fifth instar nymphs were significantly higher than those in the first and second instar nymphs. Wolbachia were scattered in all nymphal stages, but with highest intensity in the U-shaped bacteriome located in the abdomen of the nymph. Wolbachia was confined to two symmetrical organizations in the abdomen of newly emerged female and male adults. The potential mechanisms of Wolbachia infection dynamics are discussed. © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Airborne fungal spores of Alternaria, meteorological parameters and predicting variables

NASA Astrophysics Data System (ADS)

Filali Ben Sidel, Farah; Bouziane, Hassan; del Mar Trigo, Maria; El Haskouri, Fatima; Bardei, Fadoua; Redouane, Abdelbari; Kadiri, Mohamed; Riadi, Hassane; Kazzaz, Mohamed

2015-03-01

Alternaria is frequently found as airborne fungal spores and is recognized as an important cause of respiratory allergies. The aerobiological monitoring of fungal spores was performed using a Burkard volumetric spore traps. To establish predicting variables for daily and weakly spore counts, a stepwise multiple regression between spore concentrations and independent variables (meteorological parameters and lagged values from the series of spore concentrations: previous day or week concentration (Alt t - 1) and mean concentration of the same day or week in other years ( C mean)) was made with data obtained during 2009-2011. Alternaria conidia are present throughout the year in the atmosphere of Tetouan, although they show important seasonal fluctuations. The highest levels of Alternaria spores were recorded during the spring and summer or autumn. Alternaria showed maximum daily values in April, May or October depending on year. When the spore variables of Alternaria, namely C mean and Alt t - 1, and meteorological parameters were included in the equation, the resulting R 2 satisfactorily predict future concentrations for 55.5 to 81.6 % during the main spore season and the pre-peak 2. In the predictive model using weekly values, the adjusted R 2 varied from 0.655 to 0.676. The Wilcoxon test was used to compare the results from the expected values and the pre-peak spore data or weekly values for 2012, indicating that there were no significant differences between series compared. This test showed the C mean, Alt t - 1, frequency of the wind third quadrant, maximum wind speed and minimum relative humidity as the most efficient independent variables to forecast the overall trend of this spore in the air.

Dynamics of Huanglongbing-associated Bacterium Candidatus Liberibacter asiaticus in Citrus aurantifolia Swingle (Mexican Lime).

PubMed

Abel Lopez-Buenfil, Jose; Abrahan Ramirez-Pool, Jose; Ruiz-Medrano, Roberto; Del Carmen Montes-Horcasitas, Maria; Chavarin-Palacio, Claudio; Moya-Hinojosa, Jesus; Javier Trujillo-Arriaga, Francisco; Carmona, Rosalia Lira; Xoconostle-Cazares, Beatriz

2017-01-01

The bacterial disease citrus huanglongbing (HLB), associated with "Candidatus Liberibacter asiaticus" (C.Las) has severely impacted the citrus industry, causing a significant reduction in production and fruit quality. In the present study, it was monitored the C.Las population dynamics in symptomatic, HLB-positive Mexican lime trees (Citrus aurantifolia Swingle) in a tropical, citrus-producing area of Mexico. The objective of this study was to identify the dynamics of the population of huanglongbing-associated bacterium Candidatus Liberibacter asiaticus and its insect vector in Citrus aurantifolia Swingle (Mexican lime). Leaf samples were collected every 2 months over a period of 26 months for quantification of bacterial titers and young and mature leaves were collected in each season to determine preferential sites of bacterial accumulation. The proportion of living and dead bacterial cells could be determined through the use of quantitative real-time PCR in the presence of ethidium monoazide (EMA-qPCR). It was observed a lower bacterial titer at high temperatures in the infected trees relative to titers in mild weather, despite a higher accumulation of the insect vector Diaphorina citri in these conditions. This study also revealed seasonal fluctuations in the titers of bacteria in mature leaves when compared to young leaves. No statistically significant correlation between any meteorological variable, C.Las concentration and D. citri population could be drawn. Although, HLB management strategies have focused on vector control, host tree phenology may be important. The evaluation of citrus phenology, C.Las concentration, ACP population and environmental conditions provides insights into the cyclical, seasonal variations of both the HLB pathogen and its vector. These findings should help in the design of integrative HLB control strategies that take into account the accumulation of the pathogen and the presence of its vector.

Sclerenchymatous ring as a barrier to phloem feeding by Asian citrus psyllid: Evidence from electrical penetration graph and visualization of stylet pathways

USDA-ARS?s Scientific Manuscript database

Asian citrus psyllid (ACP) feeding behaviors play a significant role in the transmission of the phloem-limited Candidatus Liberibacter asiaticus (CLas) bacteria that cause citrus greening disease. Sustained phloem ingestion by ACP on CLas infected plants is very important in pathogen acquisition and...

[Activity of butenafine against ocular pathogenic filamentous fungi in vitro].

PubMed

Xu, Yan; Pang, Guang-ren; Zhao, Dong-qing; Gao, Chuan-wen; Zhou, Lu-tan; Sun, Sheng-tao; Wang, Bing-liang; Chen, Zu-ji

2010-01-01

To investigate antifungal activity of butenafine in comparison with that of natamycin, amphotericin B and fluconazole against ocular pathogenic filamentous fungi in vitro. It was an experimental study. Susceptibility tests were performed against 260 isolates of ocular pathogenic filamentous fungi by broth dilution antifungal susceptibility test of filamentous fungi approved by the Clinical and Laboratory Standards Institute (CLSI) M38-A document. The isolates included Fusarium spp. (136), Aspergillus spp. (98), Alternaria alternata (9), Curvularia lunata (3), and unusual ocular pathogens (14). Final concentration ranged from 0.008 to 16.000 mg/L for butenafine, from 0.031 to 16.000 mg/L for amphotericin B and natamycin, and from 0.5 to 256.0 mg/L for fluconazole. Following incubation at 35 degrees C for 48 h, minimal inhibitory concentration (MIC) was determined according to the CLSI M38-A document. For amphotericin B and natamycin, the MIC was defined as the lowest drug concentration that prevented any discernible growth. For butenafine and fluconazole, the MIC was defined as the lowest concentration in which an approximately 75% reduction compared to the growth of the control was observed. Candida parapsilosis ATCC22019 was used as quality control strains to validated the results. Mean MIC and MIC range, the MIC at which 50% of the isolates tested were inhibited (MIC(50)) and the MIC at which 90% of the isolates tested were inhibited (MIC(90)), were provided for all the isolates tested by using descriptive statistical analysis with the statistical SPSS package (version 13.0). MIC(90) of butenafine, natamycin, amphotericin B and fluconazole were 4, 8, 2 and 512 mg/L for Fusarium spp., respectively; 0.063, 32.000, 2.000 and 256.000 mg/L for Aspergillus spp., respectively; 0.5, 8.0, 2.0 and 128.0 mg/L for Alternaria alternate, respectively; 0.125, 2.000, 0.500 and 4.000 mg/L for Curvularia lunata, respectively; and 1, 4, 1 and 256 mg/L for unusual ocular pathogens

All roads lead to Rome: Towards understanding different avenues of tolerance to huanglongbing in citrus cultivars.

PubMed

Killiny, Nabil; Jones, Shelley E; Nehela, Yasser; Hijaz, Faraj; Dutt, Manjul; Gmitter, Frederick G; Grosser, Jude W

2018-05-18

Citrus tolerance to huanglongbing could result from tolerance to the pathogen Candidatus Liberibacter asiaticus (CLas) and/or to its vector Diaphorina citri. Field observations and greenhouse-controlled studies showed that some citrus cultivars were more tolerant than others. However, the mechanism(s) behind the tolerance has not been determined yet. Using GC-MS, we investigated the volatile organic compounds (VOCs) and the non-volatile metabolite profiles of two tolerant citrus cultivars- Australian finger lime, 'LB8-9' Sugar Belle ® mandarin hybrid, and a recently released mandarin hybrid 'Bingo'. The three were grafted onto the rootstock, Carrizo citrange. Our findings showed that the metabolomic profiles of Australian finger lime were different from that of 'LB8-9'. Finger lime was high in many amino acids and tricarboxylic acid intermediates, whereas 'LB8-9' was high in several amino acids, sugars, and sugar alcohols. 'LB8-9' was high in thymol, which is known for its strong antimicrobial activity against a panel of pathogenic bacteria. The metabolomic profiles of 'Bingo' were intensely different from the other mandarin hybrid, 'LB8-9', including a reduced thymol biosynthetic pathway and low amounts of most of the amino acids and sugar alcohols. Remarkably, 1,8-cineole (eucalyptol) was only detected in 'Bingo', indicating that eucalyptol could have feeding and ovipositional repellency against D. citri. The metabolite profiles generated for HLB-tolerant citrus species will improve the ability of citrus breeders and will allow them to take more informed decisions. Metabolomic profiling of HLB-tolerant citrus species could identify tolerance specific markers that can be introduced to other commercial citrus cultivars to improve their tolerance to HLB disease. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

The effects of citrus extract (Citrox©) on the naturally occurring microflora and inoculated pathogens, Bacillus cereus and Salmonella enterica, in a model food system and the traditional Greek yogurt-based salad Tzatziki.

PubMed

Tsiraki, Maria I; Savvaidis, Ioannis N

2016-02-01

The antimicrobial effect of citrus extract (at 1 mL/kg [TC1] and 2 mL/kg [TC2]) on the naturally occurring microflora and inoculated pathogens (Bacillus cereus and Salmonella enterica, at ca. 6 log cfu/g) in the traditional Greek yogurt-based salad Tzatziki during storage under vacuum at 4 or 10 °C was examined. We also examined the effect of citrus extract (Citrox(©)) against the two aforementioned pathogens in tryptic soy broth (TSB). Of the two treatments, TC2 yielded the lowest yeast counts, irrespective of temperature, resulting in approximately 2 (4 °C) and 3 (10 °C) log reductions on the final day of storage (70 and 30 days, respectively). Although panelists preferred the TC1-treated salad, the TC2-treated product was sensorily acceptable. Therefore, at the concentrations used, Citrox had no negative sensorial effect on the Tzatziki. During storage, the Bacillus populations in the Citrox-treated Tzatziki samples progressively decreased, showing major declines from days 12 and 28 (at 10 and 4 °C, respectively). Citrox, especially at 2 mL/kg, had a significant effect on the survival of B. cereus. S. enterica showed major declines in all untreated Tzatziki samples from day 0-70 (4 °C) and from day 0-30 (10 °C), with averages of 2.5 and 2.8 log cfu/g, respectively. The results indicate that Citrox (at 1 and 2 mL/kg) is effective, from a safety standpoint, for reducing Bacillus and Salmonella spp. in Tzatziki. In addition, 2% citrus extract also showed a higher inhibitory effect against B. cereus and S. enterica grown in TSB than 1% citrus extract. Copyright © 2015 Elsevier Ltd. All rights reserved.

Dustborne Alternaria alternata antigens in U.S. homes: Results from the National Survey of Lead and Allergens in Housing

PubMed Central

Salo, Päivi M.; Yin, Ming; Arbes, Samuel J.; Cohn, Richard D.; Sever, Michelle; Muilenberg, Michael; Burge, Harriet A.; London, Stephanie J.; Zeldin, Darryl C.

2005-01-01

Background: Alternaria alternata is one of the most common fungi associated with allergic disease. However, Alternaria exposure in indoor environments is not well characterized. Objective: The primary goals of this study were to examine the prevalence of Alternaria exposure and identify independent predictors of Alternaria antigen concentrations in U.S. homes. Methods: Data for this cross-sectional study were obtained from the National Survey of Lead and Allergens in Housing. A nationally representative sample of 831 housing units in 75 different locations throughout the U.S. completed the survey. Information on housing and household characteristics was obtained by questionnaire and environmental assessments. Concentrations of Alternaria antigens in dust collected from various indoor sites were assessed with a polyclonal anti-Alternaria antibody assay. Results: Alternaria antigens were detected in most (95-99%) of the dust samples. The geometric mean concentration, reflecting the average Alternaria concentration in homes, was 4.88 μg/g (SE=0.13 μg/g). In the multivariable linear regression analysis, the age of the housing unit, geographic region, urbanization, poverty, family race, observed mold and moisture problems, use of dehumidifier, and presence of cats and dogs were independent predictors of Alternaria antigen concentrations. Less frequent cleaning and smoking indoors also contributed to higher Alternaria antigen levels in homes. Conclusion: Exposure to Alternaria alternata antigens in U.S. homes is common. Antigen levels in homes are not only influenced by regional factors but also by residential characteristics. Preventing mold and moisture problems, avoiding smoking indoors, and regular household cleaning may help reduce exposure to Alternaria antigens indoors. PMID:16159634

Citrus diseases with global ramifications including citrus canker and huanglongbing

USDA-ARS?s Scientific Manuscript database

Although there are a number of diseases that plague citrus production worldwide, two bacterial diseases are particularly problematic. Both are of Asian origin and currently cause severe economic damage: Asiatic citrus canker (ACC) and citrus huanglongbing (HLB). Although ACC has been found in the ...

Proteomics analysis reveals novel host molecular mechanisms associated with thermotherapy of 'Ca. Liberibacter asiaticus'-infected citrus plants.

PubMed

Nwugo, Chika C; Doud, Melissa S; Duan, Yong-Ping; Lin, Hong

2016-11-14

Citrus Huanglongbing (HLB), which is linked to the bacterial pathogen 'Ca. Liberibacter asiaticus' (Las), is the most devastating disease of citrus plants, and longer-term control measures via breeding or genetic engineering have been unwieldy because all cultivated citrus species are susceptible to the disease. However, the degree of susceptibility varies among citrus species, which has prompted efforts to identify potential Las resistance/tolerance-related genes in citrus plants for application in breeding or genetic engineering programs. Plant exposure to one form of stress has been shown to serendipitously induce innate resistance to other forms of stress and a recent study showed that continuous heat treatment (40 to 42 °C) reduced Las titer and HLB-associated symptoms in citrus seedlings. The goal of the present study was to apply comparative proteomics analysis via 2-DE and mass spectrometry to elucidate the molecular processes associated with heat-induced mitigation of HLB in citrus plants. Healthy or Las-infected citrus grapefruit plants were exposed to room temperature or to continuous heat treatment of 40 °C for 6 days. An exhaustive total protein extraction process facilitated the identification of 107 differentially-expressed proteins in response to Las and/or heat treatment, which included a strong up-regulation of chaperones including small (23.6, 18.5 and 17.9 kDa) heat shock proteins, a HSP70-like protein and a ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO)-binding 60 kDa chaperonin, particularly in response to heat treatment. Other proteins that were generally down-regulated due to Las infection but up-regulated in response to heat treatment include RuBisCO activase, chlorophyll a/b binding protein, glucosidase II beta subunit-like protein, a putative lipoxygenase protein, a ferritin-like protein, and a glutathione S-transferase. The differentially-expressed proteins identified in this study highlights a premier characterization

Dual Roles of Reactive Oxygen Species and NADPH Oxidase RBOHD in an Arabidopsis-Alternaria Pathosystem1[W

PubMed Central

Pogány, Miklós; von Rad, Uta; Grün, Sebastian; Dongó, Anita; Pintye, Alexandra; Simoneau, Philippe; Bahnweg, Günther; Kiss, Levente; Barna, Balázs; Durner, Jörg

2009-01-01

Arabidopsis (Arabidopsis thaliana) NADPH oxidases have been reported to suppress the spread of pathogen- and salicylic acid-induced cell death. Here, we present dual roles of RBOHD (for respiratory burst oxidase homolog D) in an Arabidopsis-Alternaria pathosystem, suggesting either initiation or prevention of cell death dependent on the distance from pathogen attack. Our data demonstrate that a rbohD knockout mutant exhibits increased spread of cell death at the macroscopic level upon inoculation with the fungus Alternaria brassicicola. However, the cellular patterns of reactive oxygen species accumulation and cell death are fundamentally different in the AtrbohD mutant compared with the wild type. Functional RBOHD causes marked extracellular hydrogen peroxide accumulation as well as cell death in distinct, single cells of A. brassicicola-infected wild-type plants. This single cell response is missing in the AtrbohD mutant, where infection triggers spreading-type necrosis preceded by less distinct chloroplastic hydrogen peroxide accumulation in large clusters of cells. While the salicylic acid analog benzothiadiazole induces the action of RBOHD and the development of cell death in infected tissues, the ethylene inhibitor aminoethoxyvinylglycine inhibits cell death, indicating that both salicylic acid and ethylene positively regulate RBOHD and cell death. Moreover, A. brassicicola-infected AtrbohD plants hyperaccumulate ethylene and free salicylic acid compared with the wild type, suggesting negative feedback regulation of salicylic acid and ethylene by RBOHD. We propose that functional RBOHD triggers death in cells that are damaged by fungal infection but simultaneously inhibits death in neighboring cells through the suppression of free salicylic acid and ethylene levels. PMID:19726575

An Evaluation of the Genetic Diversity of Xylella fastidiosa Isolated from Diseased Citrus and Coffee in São Paulo, Brazil.

PubMed

Qin, X; Miranda, V S; Machado, M A; Lemos, E G; Hartung, J S

2001-06-01

ABSTRACT Strains of Xylella fastidiosa, isolated from sweet orange trees (Citrus sinensis) and coffee trees (Coffea arabica) with symptoms of citrus variegated chlorosis and Requeima do Café, respectively, were indistinguishable based on repetitive extragenic palindromic polymerase chain reaction (PCR) and enterobacterial repetitive intergenic consensus PCR assays. These strains were also indistinguishable with a previously described PCR assay that distinguished the citrus strains from all other strains of Xylella fastidiosa. Because we were not able to document any genomic diversity in our collection of Xylella fastidiosa strains isolated from diseased citrus, the observed gradient of increasing disease severity from southern to northern regions of São Paulo State is unlikely due to the presence of significantly different strains of the pathogen in the different regions. When comparisons were made to reference strains of Xylella fastidiosa isolated from other hosts using these methods, four groups were consistently identified consistent with the hosts and regions from which the strains originated: citrus and coffee, grapevine and almond, mulberry, and elm, plum, and oak. Independent results from random amplified polymorphic DNA (RAPD) PCR assays were also consistent with these results; however, two of the primers tested in RAPD-PCR were able to distinguish the coffee and citrus strains. Sequence comparisons of a PCR product amplified from all strains of Xylella fastidiosa confirmed the presence of a CfoI polymorphism that can be used to distinguish the citrus strains from all others. The ability to distinguish Xylella fastidiosa strains from citrus and coffee with a PCR-based assay will be useful in epidemiological and etiological studies of this pathogen.

Transmission efficiency of Xylella fastidiosa by sharpshooters (Hemiptera: Cicadellidae) in coffee and citrus.

PubMed

Marucci, Rosangela C; Lopes, João R S; Cavichioli, Rodney R

2008-08-01

Xylella fastidiosa (Wells, Raju, Hung, Weisburg, Mandelco-Paul, and Brenner) is a bacterial pathogen transmitted by several sharpshooters in two tribes of Cicadellinae (Proconiini and Cicadellini). Here, we compared the transmission efficiency of X. fastidiosa in coffee (Coffea arabica L.) and citrus [Citrus sinensis (L.) Osbeck] by Cicadellini [Bucephalogonia xanthophis (Berg) and Dilobopterus costalimai Young] and Proconiini [Homalodisca ignorata Melichar and Oncometopia facialis (Signoret)] sharpshooters that occur in both crops. At different seasons, healthy adults of each species were submitted to a 48-h acquisition access period on citrus or coffee source plants infected with X. fastidiosa isolates that cause Citrus variegated chlorosis (CVC) and Coffee leaf scorch (CLS), respectively, and then confined on healthy seedlings of the corresponding host plant for a 48-h inoculation access period. No significant effect of inoculation season was observed when comparing infection rates of citrus or coffee plants inoculated by vectors at different times of the year. In citrus, the transmission rate by single insects was significantly higher for H. ignorata (30%) in relation to B. xanthophis (5%) and O. facialis (1.1%), but there was no difference among vector species in coffee, whose transmission rates ranged from 1.2 to 7.2%. Comparing host plants, H. ignorata was more effective in transmitting X. fastidiosa to citrus (30%) in relation to coffee (2.2%), whereas the other vectors transmitted the bacterium to both hosts with similar efficiencies. Despite these variations, vector efficiency in coffee and citrus is lower than that reported in other hosts.

Use of micro-CT to elucidate details of the anatomy and feeding of the Asian citrus psyllid Diaphorina citri Kuwayama, 1908 (Insecta: Hemiptera, Liviidae)

USDA-ARS?s Scientific Manuscript database

Huanglongbing (HLB), also known as citrus greening disease, is caused by plant-infecting bacteria. The most prominent pathogen within the Americas: United States of America, Mexico, and Brazil, is Candidatus Liberibacter asiaticus, which affects plants of the Family: Rutaceae, in particularly citrus...

Expression of ß-1,3-glucanase and ß-1,4-glucanase in two potato cultivars following challenge by the fungal pathogen Alternaria solani

USDA-ARS?s Scientific Manuscript database

Early blight of potato, caused by Alternaria solani, is a ubiquitous disease in many countries around the world. We have previously found that variation in resistance phenotypes exist between two different Iranian cultivars of potato. Cultivar ‘Diamond’ is more resistant to multiple isolates of A. s...

Potential role of Alternaria and Cladosporium species in canine lymphoplasmacytic rhinitis.

PubMed

Mercier, E; Peters, I R; Billen, F; Battaille, G; Clercx, C; Day, M J; Peeters, D

2013-04-01

To evaluate the possible role of Alternaria and Cladosporium species in the pathogenesis of canine lymphoplasmacytic rhinitis by comparing the amount of specific fungal DNA in nasal mucosal biopsies between dogs without nasal neoplasia and those with lymphoplasmacytic rhinitis or nasal neoplasia. Quantitative real-time polymerase chain reaction (qPCR) assays detecting DNA from Alternaria and Cladosporium fungi were applied to nasal mucosal biopsies collected from dogs with lymphoplasmacytic rhinitis (n = 8), dogs with nasal neoplasia (n = 10) and control animals (n = 10). A copy number for each sample was calculated using a standard curve of known copy number and differences amongst groups were assessed using Kruskal-Wallis tests. No significant difference was found between the groups. Low levels of Alternaria DNA (10-100 copies/PCR) were detected in one sample; very low levels of DNA (<10 copies/qPCR) were detected in 6 samples, and 21 samples were negative. Low levels of Cladosporium DNA were detected in 2 samples; very low levels of DNA in 18; and 8 were negative. Results of this study reveal that Alternaria and Cladosporium species are part of the canine nasal flora, and that these fungi are probably not involved in the pathogenesis of lymphoplasmacytic rhinitis. © 2013 British Small Animal Veterinary Association.

Temporal progression of 'Candidatus Liberibacter asiaticus' infection in citrus and acquisition efficiency by Diaphorina citri.

PubMed

Coletta-Filho, Helvecio D; Daugherty, Matthew P; Ferreira, Cléderson; Lopes, João R S

2014-04-01

Over the last decade, the plant disease huanglongbing (HLB) has emerged as a primary threat to citrus production worldwide. HLB is associated with infection by phloem-limited bacteria ('Candidatus Liberibacter' spp.) that are transmitted by the Asian citrus psyllid, Diaphorina citri. Transmission efficiency varies with vector-related aspects (e.g., developmental stage and feeding periods) but there is no information on the effects of host-pathogen interactions. Here, acquisition efficiency of 'Candidatus Liberibacter asiaticus' by D. citri was evaluated in relation to temporal progression of infection and pathogen titer in citrus. We graft inoculated sweet orange trees with 'Ca. L. asiaticus'; then, at different times after inoculation, we inspected plants for HLB symptoms, measured bacterial infection levels (i.e., titer or concentration) in plants, and measured acquisition by psyllid adults that were confined on the trees. Plant infection levels increased rapidly over time, saturating at uniformly high levels (≈10(8) copy number of 16S ribosomal DNA/g of plant tissue) near 200 days after inoculation-the same time at which all infected trees first showed disease symptoms. Pathogen acquisition by vectors was positively associated with plant infection level and time since inoculation, with acquisition occurring as early as the first measurement, at 60 days after inoculation. These results suggest that there is ample potential for psyllids to acquire the pathogen from trees during the asymptomatic phase of infection. If so, this could limit the effectiveness of tree rouging as a disease management tool and would likely explain the rapid spread observed for this disease in the field.

Comparative Genomic Analysis of Xanthomonas axonopodis pv. citrumelo F1, Which Causes Citrus Bacterial Spot Disease, and Related Strains Provides Insights into Virulence and Host Specificity ▿ #

PubMed Central

Jalan, Neha; Aritua, Valente; Kumar, Dibyendu; Yu, Fahong; Jones, Jeffrey B.; Graham, James H.; Setubal, João C.; Wang, Nian

2011-01-01

Xanthomonas axonopodis pv. citrumelo is a citrus pathogen causing citrus bacterial spot disease that is geographically restricted within the state of Florida. Illumina, 454 sequencing, and optical mapping were used to obtain a complete genome sequence of X. axonopodis pv. citrumelo strain F1, 4.9 Mb in size. The strain lacks plasmids, in contrast to other citrus Xanthomonas pathogens. Phylogenetic analysis revealed that this pathogen is very close to the tomato bacterial spot pathogen X. campestris pv. vesicatoria 85-10, with a completely different host range. We also compared X. axonopodis pv. citrumelo to the genome of citrus canker pathogen X. axonopodis pv. citri 306. Comparative genomic analysis showed differences in several gene clusters, like those for type III effectors, the type IV secretion system, lipopolysaccharide synthesis, and others. In addition to pthA, effectors such as xopE3, xopAI, and hrpW were absent from X. axonopodis pv. citrumelo while present in X. axonopodis pv. citri. These effectors might be responsible for survival and the low virulence of this pathogen on citrus compared to that of X. axonopodis pv. citri. We also identified unique effectors in X. axonopodis pv. citrumelo that may be related to the different host range as compared to that of X. axonopodis pv. citri. X. axonopodis pv. citrumelo also lacks various genes, such as syrE1, syrE2, and RTX toxin family genes, which were present in X. axonopodis pv. citri. These may be associated with the distinct virulences of X. axonopodis pv. citrumelo and X. axonopodis pv. citri. Comparison of the complete genome sequence of X. axonopodis pv. citrumelo to those of X. axonopodis pv. citri and X. campestris pv. vesicatoria provides valuable insights into the mechanism of bacterial virulence and host specificity. PMID:21908674

Application of ultraviolet-C light on oranges for the inactivation of postharvest wound pathogens

USDA-ARS?s Scientific Manuscript database

Germicidal effects of ultraviolet-C (UV-C) light on the postharvest wound pathogens of citrus fruits namely Penicillium digitatum and Penicillium italicum were investigated. P. digitatum and P. italicum spores were inoculated (4.00 – 4.50 log cfu/ orange) onto Washington navel oranges (Citrus sinens...

Citrus leprosis research update

USDA-ARS?s Scientific Manuscript database

Citrus leprosis is one of the oldest citrus diseases, but is also one of the most important emerging citrus diseases in South and Central America, and it is apparently spreading northward towards the U.S. Research in our labs and by others has shown that citrus leprosis disease is caused by a compl...

40 CFR 180.1256 - Alternaria destruens strain 059; exemption from the requirement of a tolerance.

Code of Federal Regulations, 2011 CFR

2011-07-01

... RESIDUES IN FOOD Exemptions From Tolerances § 180.1256 Alternaria destruens strain 059; exemption from the... the microbial pesticide Alternaria destruens Strain 059 when used in or on all raw agricultural...

40 CFR 180.1256 - Alternaria destruens strain 059; exemption from the requirement of a tolerance.

Code of Federal Regulations, 2010 CFR

2010-07-01

... RESIDUES IN FOOD Exemptions From Tolerances § 180.1256 Alternaria destruens strain 059; exemption from the... the microbial pesticide Alternaria destruens Strain 059 when used in or on all raw agricultural...

A Novel Alternaria Species Isolated from Peucedanum japonicum in Korea

PubMed Central

Deng, Jian Xin; Cho, Hye Sun; Paul, Narayan Chandra; Lee, Hyang Burm

2014-01-01

We isolated and examined a new Alternaria sp., which causes leaf spots on Peucedanum japonicum in Korea, by using molecular and morphological methods. Phylogenetic analysis based on a combined internal transcribed spacer region analysis and two protein-coding genes (gpd and Alt a1) demonstrated that the causal fungus was most closely related to A. cinerariae and A. sonchi, and relevant to A. brassicae. However, conidial morphology indicated that it is a novel species within the genus Alternaria, and therefore we have assigned the fungus a new name in this study. PMID:24808728

Multiple phytohormone signalling pathways modulate susceptibility of tomato plants to Alternaria alternata f. sp. lycopersici

PubMed Central

Jia, Chengguo; Zhang, Liping; Wang, Qiaomei

2013-01-01

Three phytohormone molecules – ethylene (ET), jasmonic acid (JA) and salicylic acid (SA) – play key roles in mediating disease response to necrotrophic fungal pathogens. This study investigated the roles of the ET, JA, and SA pathways as well as their crosstalk during the interaction between tomato (Solanum lycopersicum) plants and a necrotrophic fungal pathogen Alternaria alternata f. sp. lycopersici (AAL). Both the ET and JASMONIC ACID INSENSITIVE1 (JAI1) receptor-dependent JA signalling pathways are necessary for susceptibility, while SA response promotes resistance to AAL infection. In addition, the role of JA in susceptibility to AAL is partly dependent on ET biosynthesis and perception, while the SA pathway enhances resistance to AAL and antagonizes the ET response. Based on these results, it is proposed that ET, JA, and SA each on their own can influence the susceptibility of tomato to AAL. Furthermore, the functions of JA and SA in susceptibility to the pathogen are correlated with the enhanced or decreased action of ET, respectively. This study has revealed the functional relationship among the three key hormone pathways in tomato defence against AAL. PMID:23264518

Production of transgenic citrus resistant to citrus canker and Huanglongbing diseases

USDA-ARS?s Scientific Manuscript database

Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the U.S. citrus industry. There are no proven strategies to eliminate HLB disease and no cultivars identified with strong HLB resistance. Citrus canker is also an economically import...

Effect of eosinophils activated with Alternaria on the production of extracellular matrix from nasal fibroblasts.

PubMed

Shin, Seung-Heon; Ye, Mi-Kyung; Choi, Sung-Yong; Kim, Yee-Hyuk

2016-06-01

Eosinophils and fibroblasts are known to play major roles in the pathogenesis of nasal polyps. Fungi are commonly found in nasal secretion and are associated with airway inflammation. To investigate whether activated eosinophils by airborne fungi can influence the production of extracellular matrix (ECM) from nasal fibroblasts. Inferior turbinate and nasal polyp fibroblasts were stimulated with Alternaria or Aspergillus, respectively, for 24 hours and ECM messenger RNA (mRNA) and protein expressions were measured. Eosinophils isolated from healthy volunteers were stimulated with Alternaria or Aspergillus for 4 hours then superoxide, eosinophil peroxidase, and transforming growth factor β1 were measured. Then activated eosinophils were cocultured with nasal fibroblasts for 24 hours, and ECM mRNA expressions were measured. Alternaria strongly enhanced ECM mRNA expression and protein production from nasal fibroblasts. Alternaria also induced the production of superoxide, eosinophil peroxidase, and transforming growth factor β1 from eosinophils, and activated eosinophils enhanced ECM mRNA expression when they were cocultured without the Transwell insert system. Eosinophils activated with Alternaria enhanced ECM mRNA expression from nasal polyp fibroblasts. Alternaria plays an important role in tissue fibrosis in the pathogenesis of nasal polyps by directly or indirectly influencing the production of ECM from nasal fibroblasts. Copyright © 2016 American College of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Zinc treatment increases the titre of 'Candidatus Liberibacter asiaticus' in huanglongbing-affected citrus plants while affecting the bacterial microbiomes.

PubMed

Zhang, M Q; Guo, Y; Powell, C A; Doud, M S; Yang, C Y; Zhou, H; Duan, Y P

2016-06-01

Huanglongbing (HLB)-affected citrus often display zinc deficiency symptoms. In this study, supplemental zinc was applied to citrus to determine its effect on Candidatus Liberibacter asiaticus (Las) titre, HLB symptoms, and leaf microbiome. HLB-affected citrus were treated with various amounts of zinc. The treatments promoted Las growth and affected microbiomes in citrus leaves. Phylochip(™) -based results indicated that 5475 of over 50 000 known Operational Taxonomic Units (OTUs) in 52 phyla were detected in the midribs of HLB-affected citrus, of which Proteobacteria was the most abundant, followed by Firmicutes and Actinobacteria. In comparison, the microbiomes of zinc-treated diseased plants had overall more OTUs with higher amounts of Proteobacteria, but decreased percentages of Firmicutes and Actinobacteria. In addition, more OTUs of siderophore-producing bacteria were present. Only zinc-sensitive Staphylococcaceae had higher OTU's in the diseased plants without zinc treatments. Although HLB-affected citrus appear zinc deficient, zinc amendments increased the pathogen levels and shifted the microbiome. HLB is currently the most devastating disease of citrus worldwide. Zinc is often applied to HLB-affected citrus due to zinc deficiency symptoms. This study provided new insights into the potential effects of zinc on HLB and the microbial ecology of citrus. © 2016 The Society for Applied Microbiology.

Development of a polymerase chain reaction-based assay for the detection of Alternaria fungal contamination in food products.

PubMed

Zur, G; Hallerman, E M; Sharf, R; Kashi, Y

1999-10-01

Alternaria sp. are important fungal contaminants of vegetable, fruit, and grain products, including Alternaria alternata, a contaminant of tomato products. To date, the Howard method, based on microscopic observation of fungal filaments, has been the standard examination for inspection of tomato products. We report development of a polymerase chain reaction (PCR)-based method for detection of Alternaria DNA. PCR primers were designed to anneal to the internal transcribed regions ITS1 and ITS2 of the 5.8S rRNA gene of Alternaria but not to other microbial or tomato DNA. We demonstrate use of the PCR assay to detect Alternaria DNA in experimentally infested and commercially obtained tomato sauce and tomato powder. Use of the PCR method offers a rapid and sensitive assay for the presence of Alternaria DNA in tomato products. The apparent breakdown of DNA in tomato sauce may limit the utility of the assay to freshly prepared products. The assay for tomato powder is not affected by storage time.

The study of Bacteriocin of Pseudomonas fluorescens and Citrus limon effects against Propionibacterium acnes and Staphylococcus epidermidis in acne patients

NASA Astrophysics Data System (ADS)

Ahmed, Mais E.

2018-05-01

Research was carried out on the antibacterial effect of (Citrus limon) juice on Acnevulgaris. Samples were obtained from individuals with pimples by swabbing their faces. Natural substances that derive from plants are promising to treat disease cause Acnevulgaris, the study in vitro biological activity of the juice, as well as bacterocin cultivated and fruits was investigated on two strains of bacteria (Propionibacterium acnes, Staphylococcus epidermidis). The new antimicrobial (bacteriocin and Citrus juice) is an ongoing search. This study used juice at different concentrations at (20%, 30%, 40%, 60%, 80% and 100%). The bacteriocin produced from local P. fluorescens isolates from wound infection and majority of isolates were found to produce crude bacteriocin were (P1 and P2) in Pseudomonas agar at 37°C for 24 hrs. Crude bacteriocin and Citrus limon juice against some pathogenic skin bacteria was find to be effective juice Citrus limon aganist S. epidermidis at 100% Concentrations with a range of inhibition zone (18) mm. The isolates of P. fluorescens (P2) was positive as producer of bacteriocin with a wide inhibition growth against gram positive pathogenic bacteria with a range between (10-12) mm.

Emerging Fusarium and Alternaria Mycotoxins: Occurrence, Toxicity and Toxicokinetics

PubMed Central

Croubels, Siska; Devreese, Mathias; Antonissen, Gunther

2017-01-01

Emerging Fusarium and Alternaria mycotoxins gain more and more interest due to their frequent contamination of food and feed, although in vivo toxicity and toxicokinetic data are limited. Whereas the Fusarium mycotoxins beauvericin, moniliformin and enniatins particularly contaminate grain and grain-based products, Alternaria mycotoxins are also detected in fruits, vegetables and wines. Although contamination levels are usually low (µg/kg range), higher contamination levels of enniatins and tenuazonic acid may occasionally occur. In vitro studies suggest genotoxic effects of enniatins A, A1 and B1, beauvericin, moniliformin, alternariol, alternariol monomethyl ether, altertoxins and stemphyltoxin-III. Furthermore, in vitro studies suggest immunomodulating effects of most emerging toxins and a reproductive health hazard of alternariol, beauvericin and enniatin B. More in vivo toxicity data on the individual and combined effects of these contaminants on reproductive and immune system in both humans and animals is needed to update the risk evaluation by the European Food Safety Authority. Taking into account new occurrence data for tenuazonic acid, the complete oral bioavailability, the low total body clearance in pigs and broiler chickens and the limited toxicity data, a health risk cannot be completely excluded. Besides, some less known Alternaria toxins, especially the genotoxic altertoxins and stemphyltoxin III, should be incorporated in risk evaluation as well. PMID:28718805

Antimicrobial effects of citrus sinensis peel extracts against periodontopathic bacteria: an in vitro study.

PubMed

Hussain, Khaja Amjad; Tarakji, Bassel; Kandy, Binu Purushothaman Panar; John, Jacob; Mathews, Jacob; Ramphul, Vandana; Divakar, Darshan Devang

2015-01-01

Use of plant extracts and phytochemicals with known antimicrobial properties may have great significance in therapeutic treatments. To assess the in vitro antimicrobial potential and also determine the minimum inhibitory concentration (MIC) of Citrus sinensis peel extracts with a view of searching a novel extract as a remedy for periodontal pathogens. Aqueous and ethanol (cold and hot) extracts prepared from peel of Citrus sinensis were screened for in vitro antimicrobial activity against Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia, using agar well diffusion method. The lowest concentration of every extract considered as the minimal inhibitory concentration (MIC) values were determined for both test organisms. Confidence level and level of significance were set at 95% and 5% respectively. Prevotella intermedia and Porphyromonas gingivalis were resistant to aqueous extracts while Aggregatibacter actinomycetemcomitans was inhibited at very high cncentrations. Hot ethanolic extracts showed significantly higher zone of inhibition than cold ethanolic extract. Minimum inhibitory concentration of hot and cold ethanolic extracts of Citrus sinensis peel ranged between 12-15 mg/ml against all three periodontal pathogens. Both extracts were found sensitive and contain compounds with therapeutic potential. Nevertheless, clinical trials on the effect of these plants are essential before advocating large-scale therapy.

Field ID guide to citrus relative hosts of Asian citrus psyllid & Huanglongbing

USDA-ARS?s Scientific Manuscript database

The Rutaceae family of plants includes not only species within the genus Citrus, but also several other genera and species that may not be easily recognized as having any relationship to citrus at all. However, many of these citrus relatives are used for ornamental, culinary, or religious purposes. ...

All five host-range variants of Xanthomonas citri carry one pthA homolog with 17.5 repeats that determines pathogenicity on citrus, but none determine host-range variation.

PubMed

Al-Saadi, Abdulwahid; Reddy, Joseph D; Duan, Yong P; Brunings, Asha M; Yuan, Qiaoping; Gabriel, Dean W

2007-08-01

Citrus canker disease is caused by five groups of Xanthomonas citri strains that are distinguished primarily by host range: three from Asia (A, A*, and A(w)) and two that form a phylogenetically distinct clade and originated in South America (B and C). Every X. citri strain carries multiple DNA fragments that hybridize with pthA, which is essential for the pathogenicity of wide-host-range X. citri group A strain 3213. DNA fragments that hybridized with pthA were cloned from a representative strain from all five groups. Each strain carried one and only one pthA homolog that functionally complemented a knockout mutation of pthA in 3213. Every complementing homolog was of identical size to pthA and carried 17.5 nearly identical, direct tandem repeats, including three new genes from narrow-host-range groups C (pthC), A(w) (pthAW), and A* (pthA*). Every noncomplementing paralog was of a different size; one of these was sequenced from group A* (pthA*-2) and was found to have an intact promoter and full-length reading frame but with 15.5 repeats. None of the complementing homologs nor any of the noncomplementing paralogs conferred avirulence to 3213 on grapefruit or suppressed avirulence of a group A* strain on grapefruit. A knockout mutation of pthC in a group C strain resulted in loss of pathogenicity on lime, but the strain was unaffected in ability to elicit an HR on grapefruit. This pthC- mutant was fully complemented by pthA, pthB, or pthC. Analysis of the predicted amino-acid sequences of all functional pthA homologs and nonfunctional paralogs indicated that the specific sequence of the 17th repeat may be essential for pathogenicity of X. citri on citrus.

Rapid differentiation of citrus Hop stunt viroid variants by use of real-time RT-PCR and high resolution melting analysis

USDA-ARS?s Scientific Manuscript database

The RNA genome of Hop stunt viroid (HSVd) contains five to six nucleotides in a variable (V) domain, called the cachexia expression motif, which is associated with pathogenic and non-pathogenic variants in citrus. Current methods to differentiate HSVd variants rely on lengthy greenhouse biological i...

Collection and chemical composition of phloem sap from Citrus sinensis L. Osbeck (sweet orange).

PubMed

Hijaz, Faraj; Killiny, Nabil

2014-01-01

Through utilizing the nutrient-rich phloem sap, sap feeding insects such as psyllids, leafhoppers, and aphids can transmit many phloem-restricted pathogens. On the other hand, multiplication of phloem-limited, uncultivated bacteria such as Candidatus Liberibacter asiaticus (CLas) inside the phloem of citrus indicates that the sap contains all the essential nutrients needed for the pathogen growth. The phloem sap composition of many plants has been studied; however, to our knowledge, there is no available data about citrus phloem sap. In this study, we identified and quantified the chemical components of phloem sap from pineapple sweet orange. Two approaches (EDTA enhanced exudation and centrifugation) were used to collect phloem sap. The collected sap was derivatized with methyl chloroformate (MCF), N-methyl-N- [tert-butyl dimethylsilyl]-trifluroacetamide (MTBSTFA), or trimethylsilyl (TMS) and analyzed with GC-MS revealing 20 amino acids and 8 sugars. Proline, the most abundant amino acid, composed more than 60% of the total amino acids. Tryptophan, tyrosine, leucine, isoleucine, and valine, which are considered essential for phloem sap-sucking insects, were also detected. Sucrose, glucose, fructose, and inositol were the most predominant sugars. In addition, seven organic acids including succinic, fumaric, malic, maleic, threonic, citric, and quinic were detected. All compounds detected in the EDTA-enhanced exudate were also detected in the pure phloem sap using centrifugation. The centrifugation technique allowed estimating the concentration of metabolites. This information expands our knowledge about the nutrition requirement for citrus phloem-limited bacterial pathogen and their vectors, and can help define suitable artificial media to culture them.

Collection and Chemical Composition of Phloem Sap from Citrus sinensis L. Osbeck (Sweet Orange)

PubMed Central

Hijaz, Faraj; Killiny, Nabil

2014-01-01

Through utilizing the nutrient-rich phloem sap, sap feeding insects such as psyllids, leafhoppers, and aphids can transmit many phloem-restricted pathogens. On the other hand, multiplication of phloem-limited, uncultivated bacteria such as Candidatus Liberibacter asiaticus (CLas) inside the phloem of citrus indicates that the sap contains all the essential nutrients needed for the pathogen growth. The phloem sap composition of many plants has been studied; however, to our knowledge, there is no available data about citrus phloem sap. In this study, we identified and quantified the chemical components of phloem sap from pineapple sweet orange. Two approaches (EDTA enhanced exudation and centrifugation) were used to collect phloem sap. The collected sap was derivatized with methyl chloroformate (MCF), N-methyl-N- [tert-butyl dimethylsilyl]-trifluroacetamide (MTBSTFA), or trimethylsilyl (TMS) and analyzed with GC-MS revealing 20 amino acids and 8 sugars. Proline, the most abundant amino acid, composed more than 60% of the total amino acids. Tryptophan, tyrosine, leucine, isoleucine, and valine, which are considered essential for phloem sap-sucking insects, were also detected. Sucrose, glucose, fructose, and inositol were the most predominant sugars. In addition, seven organic acids including succinic, fumaric, malic, maleic, threonic, citric, and quinic were detected. All compounds detected in the EDTA-enhanced exudate were also detected in the pure phloem sap using centrifugation. The centrifugation technique allowed estimating the concentration of metabolites. This information expands our knowledge about the nutrition requirement for citrus phloem-limited bacterial pathogen and their vectors, and can help define suitable artificial media to culture them. PMID:25014027

Characterization of Mycosphaerellaceae species associated with citrus greasy spot in Panama and Spain.

PubMed

Aguilera-Cogley, Vidal Antonio; Berbegal, Mónica; Català, Santiago; Brentu, Francis Collison; Armengol, Josep; Vicent, Antonio

2017-01-01

Greasy spot of citrus, caused by Zasmidium citri-griseum (= Mycosphaerella citri), is widely distributed in the Caribbean Basin, inducing leaf spots, premature defoliation, and yield loss. Greasy spot-like symptoms were frequently observed in humid citrus-growing regions in Panama as well as in semi-arid areas in Spain, but disease aetiology was unknown. Citrus-growing areas in Panama and Spain were surveyed and isolates of Mycosphaerellaceae were obtained from citrus greasy spot lesions. A selection of isolates from Panama (n = 22) and Spain (n = 16) was assembled based on their geographical origin, citrus species, and affected tissue. The isolates were characterized based on multi-locus DNA (ITS and EF-1α) sequence analyses, morphology, growth at different temperatures, and independent pathogenicity tests on the citrus species most affected in each country. Reference isolates and sequences were also included in the analysis. Isolates from Panama were identified as Z. citri-griseum complex, and others from Spain attributed to Amycosphaerella africana. Isolates of the Z. citri-griseum complex had a significantly higher optimal growth temperature (26.8°C) than those of A. africana (19.3°C), which corresponded well with their actual biogeographical range. The isolates of the Z. citri-griseum complex from Panama induced typical greasy spot symptoms in 'Valencia' sweet orange plants and the inoculated fungi were reisolated. No symptoms were observed in plants of the 'Ortanique' tangor inoculated with A. africana. These results demonstrate the presence of citrus greasy spot, caused by Z. citri-griseum complex, in Panama whereas A. africana was associated with greasy spot-like symptoms in Spain.

Characterization of Mycosphaerellaceae species associated with citrus greasy spot in Panama and Spain

PubMed Central

Aguilera-Cogley, Vidal Antonio; Berbegal, Mónica; Català, Santiago; Brentu, Francis Collison; Armengol, Josep

2017-01-01

Greasy spot of citrus, caused by Zasmidium citri-griseum (= Mycosphaerella citri), is widely distributed in the Caribbean Basin, inducing leaf spots, premature defoliation, and yield loss. Greasy spot-like symptoms were frequently observed in humid citrus-growing regions in Panama as well as in semi-arid areas in Spain, but disease aetiology was unknown. Citrus-growing areas in Panama and Spain were surveyed and isolates of Mycosphaerellaceae were obtained from citrus greasy spot lesions. A selection of isolates from Panama (n = 22) and Spain (n = 16) was assembled based on their geographical origin, citrus species, and affected tissue. The isolates were characterized based on multi-locus DNA (ITS and EF-1α) sequence analyses, morphology, growth at different temperatures, and independent pathogenicity tests on the citrus species most affected in each country. Reference isolates and sequences were also included in the analysis. Isolates from Panama were identified as Z. citri-griseum complex, and others from Spain attributed to Amycosphaerella africana. Isolates of the Z. citri-griseum complex had a significantly higher optimal growth temperature (26.8°C) than those of A. africana (19.3°C), which corresponded well with their actual biogeographical range. The isolates of the Z. citri-griseum complex from Panama induced typical greasy spot symptoms in ‘Valencia’ sweet orange plants and the inoculated fungi were reisolated. No symptoms were observed in plants of the ‘Ortanique’ tangor inoculated with A. africana. These results demonstrate the presence of citrus greasy spot, caused by Z. citri-griseum complex, in Panama whereas A. africana was associated with greasy spot-like symptoms in Spain. PMID:29236789

Cost-effectiveness of various methods of diagnosing hypersensitivity to Alternaria.

PubMed

Escudero, A I; Sánchez-Guerrero, I M; Mora, A M; Soriano, V; López, J D; García, F J; Negro, J M; Hernández, J; Pagán, J A

1993-01-01

This study was undertaken for two reasons: 1) It is more difficult to diagnose hypersensitivity to molds than to other allergens, so an evaluation of diagnostic tests was needed. 2) Alternaria is the principal cause of mold sensitization in our area. Sixty-six patients (20 +/- 4 years) were selected and divided into two groups. Group A was made up of patients with rhinitis and/or asthma due to Alternaria sensitization. Group B consisted of patients sensitized to other allergens and patients with nonrespiratory allergic disorders. Skin tests (prick and intradermal), challenge tests (conjunctival, nasal, and bronchial), and specific IgE determination were performed for all patients. A biologically standardized extract of Alternaria tenuis (Alergia e Inmunología Abelló, S. A., Madrid, Spain) obtained from a single batch was used for all tests. Our diagnostic criterion was a clinical history of rhinitis or asthma that coincided with the results of nasal/bronchial challenge. The diagnostic value of the other tests was compared to this criterion. In the group of rhinitic patients, skin tests and conjunctival challenge were more sensitive than specific IgE determination. In asthmatic patients, the most sensitive techniques were nasal and conjunctival challenges, followed by prick and intradermal skin tests, and, lastly, serum specific IgE determination. When rhinitis and asthma were considered jointly, the most sensitive test was conjunctival challenge, followed by skin-prick and intradermal tests. All tests had the same specificity, regardless of disorder. Nasal challenge was positive in all patients. Skin tests are easy to perform, cheap, non-traumatic for the patient, and sufficiently specific and sensitive for the diagnosis of Alternaria hypersensitivity.(ABSTRACT TRUNCATED AT 250 WORDS)

The WRKY Transcription Factor Family in Citrus: Valuable and Useful Candidate Genes for Citrus Breeding.

PubMed

Ayadi, M; Hanana, M; Kharrat, N; Merchaoui, H; Marzoug, R Ben; Lauvergeat, V; Rebaï, A; Mzid, R

2016-10-01

WRKY transcription factors belong to a large family of plant transcriptional regulators whose members have been reported to be involved in a wide range of biological roles including plant development, adaptation to environmental constraints and response to several diseases. However, little or poor information is available about WRKY's in Citrus. The recent release of completely assembled genomes sequences of Citrus sinensis and Citrus clementina and the availability of ESTs sequences from other citrus species allowed us to perform a genome survey for Citrus WRKY proteins. In the present study, we identified 100 WRKY members from C. sinensis (51), C. clementina (48) and Citrus unshiu (1), and analyzed their chromosomal distribution, gene structure, gene duplication, syntenic relation and phylogenetic analysis. A phylogenetic tree of 100 Citrus WRKY sequences with their orthologs from Arabidopsis has distinguished seven groups. The CsWRKY genes were distributed across all ten sweet orange chromosomes. A comprehensive approach and an integrative analysis of Citrus WRKY gene expression revealed variable profiles of expression within tissues and stress conditions indicating functional diversification. Thus, candidate Citrus WRKY genes have been proposed as potentially involved in fruit acidification, essential oil biosynthesis and abiotic/biotic stress tolerance. Our results provided essential prerequisites for further WRKY genes cloning and functional analysis with an aim of citrus crop improvement.

Does citrus leaf miner impair hydraulics and fitness of citrus host plants?

PubMed

Raimondo, Fabio; Trifilò, Patrizia; Gullo, Maria A Lo

2013-12-01

Gas exchange and hydraulic features were measured in leaves of three different Citrus species (Citrus aurantium L., Citrus limon L., Citrus  ×  paradisii Macfad) infested by Phyllocnistis citrella Staiton, with the aim to quantify the impact of this pest on leaf hydraulics and, ultimately, on plant fitness. Infested leaves were characterized by the presence on the leaf blade of typical snake-shaped mines and, in some cases, of a crumpled leaf blade. Light microscopy showed that leaf crumpling was induced by damage to the cuticular layer. In all three Citrus species examined: (a) the degree of infestation did not exceed 10% of the total surface area of infested plants; (b) control and infested leaves showed similar values of minimum diurnal leaf water potential, leaf hydraulic conductance and functional vein density; and (c) maximum diurnal values of stomatal conductance to water vapour, transpiration rate and photosynthetic rate (An) were similar in both control leaves and the green areas of infested leaves. A strong reduction of An was recorded only in mined leaf areas. Our data suggest that infestation with P. citrella does not cause conspicuous plant productivity reductions in young Citrus plants, at least not in the three Citrus species studied here.

Secondary metabolite profiling of Alternaria dauci, A. porri, A. solani, and A. tomatophila.

PubMed

Andersen, Birgitte; Dongo, Anita; Pryor, Barry M

2008-02-01

Chemotaxonomy (secondary metabolite profiling) has been shown to be of great value in the classification and differentiation in Ascomycota. However, few studies have investigated the use of metabolite production for classification and identification purposes of plant pathogenic Alternaria species. The purpose of the present study was to describe the methodology behind metabolite profiling in chemotaxonomy using A. dauci, A. porri, A. solani, and A. tomatophila strains as examples of the group. The results confirmed that A. dauci, A. solani, and A. tomatophila are three distinct species each with their own specific metabolite profiles, and that A. solani and A. tomatophila both produce altersolanol A, altertoxin I, and macrosporin. By using automated chemical image analysis and other multivariate statistic analyses, three sets of species-specific metabolites could be selected, one each for A. dauci, A. solani, and A. tomatophila.

Transformation of sweet orange [Citrus sinensis (L.) Osbeck] with pthA-nls for acquiring resistance to citrus canker disease.

PubMed

Yang, Li; Hu, Chunhua; Li, Na; Zhang, Jiayin; Yan, Jiawen; Deng, Ziniu

2011-01-01

The COOH terminal of pthA encoding three nuclear localizing signals (NLS) was amplified by polymerase chain reaction (PCR) from the plasmid of Xanthomonas axonopodis pv. citri, the pathogen of citrus canker disease. Then the sense and antisense strands of the nls were cloned into pBI121 vector. pthA-nls driven by the CaMV35 s promoter was transferred into sweet orange via Agrobacterium -mediated transformation. Successful integration was confirmed by PCR and Southern blotting, and 12 sense-nls (nls (+)) and 9 antisense-nls (nls (-)) transgenic clones were obtained. The expression of nls fragment was analyzed by RT-PCR, Real time q-PCR and Western blotting, in which the specific NLS protein was detected only in nls (+) transgenic clones. In an in vitro assay, when pin-puncture inoculation was performed with 2.5 × 10(7) cfu/ml of bacterial solution, the nls (+) transgenic clones showed no typical lesion development, while typical symptoms were observed in the wild types and the nls (-) transgenic clones. In vivo assay results indicated that the nls (+) transgenic clones showed less disease incidence, in comparison with the wild types and the nls (-) transgenic clones, when pin-puncture inoculation was performed with 10(4)-10(5) cfu/ml. The minimum disease incidence was 23.3% for 'Sucarri' sweet orange and 33.3% for 'Bingtang' sweet orange. When 10(4)-10(7) cfu/ml of pathogen was spray inoculated, the nls (+) transgenic clones did not show any symptom, and even the concentration raised to 10(9) cfu/ml, the disease incidence was 20-80%, while the wild types and the nls (-) transgenic clones had 100% disease development with whatever concentration of inoculum. Two transgenic clones were confirmed to be resistant to citrus canker disease in the repeated inoculation. The results suggested that the transformation of nls sense strands may offer an effective way to acquire resistance to citrus canker disease.

Development of selective media for the isolation and enumeration of Alternaria species from soil and plant debris.

PubMed

Hong, Soon Gyu; Pryor, Barry M

2004-07-01

A new semi-selective medium, acidified weak potato-dextrose agar (AWPDA) with Mertect (active ingredient: thiabendazole), was developed for the isolation and enumeration of Alternaria species from samples of soil and plant debris. The medium was selected based on growth inhibition tests against Alternaria and several other commonly encountered saprobic fungi utilizing three antifungal agents, Botran (active ingredient: dichloran), Bayleton (active ingredient: triadimefon), and Mertect, and two basal media, acidified potato-dextrose agar (APDA) and AWPDA. Botran inhibited growth of Rhizopus stolonifer moderately, but had little effect on Cladosporium cladosporoides, Fusarium oxysporum, Penicillium chrysogenum, or Trichoderma harzianum. Bayleton inhibited growth of R. stolonifer and C. cladosporoides severely, and inhibited growth of F. oxysporum, P. chrysogenum, and T. harzianum moderately. Mertect inhibited growth of C. cladosporoides, F. oxysporum, P. chrysogenum, and T. harzianum completely, but had little or moderate effect on R. stolonifer. All three antifungal agents inhibited growth of Alternaria species slightly or moderately. The combination of Bayleton and Mertect inhibited growth of all fungi severely. A comparison of recovery rates of Alternaria from soil and plant debris samples on AWPDA with Mertect and weak potato-dextrose agar (WPDA) revealed that Alternaria spp. accounted for 63.6%-81.0% of recovered fungal isolates on AWPDA with Mertect as compared to 0.6%-2.7% of recovered isolates on WPDA. The AWPDA medium with Mertect exhibited superior selective growth of Alternaria species from samples of soil and plant debris, and will be useful in studies where the recovery and enumeration of Alternaria species is necessary.

Dynamics of 'Candidatus Liberibacter asiaticus' Colonization of New Growth of Citrus.

PubMed

Hilf, Mark E; Luo, Weiqi

2018-05-14

'Candidatus Liberibacter asiaticus' is a phloem-colonizing intracellular bacterial pathogen of citrus associated with the disease huanglongbing. A study of patterns of colonization and bacterial population growth in new growth of different citrus types was conducted by pruning infected citron, sweet orange, sour orange, mandarin, citrange and Citrus macrophylla trees to force the growth of axillary and adventitious shoots. The first three leaves on newly emerged shoots were collected at 30, 60 and 90 days to assess colonization and population growth of 'Ca. L. asiaticus' using real time PCR (qPCR). Single trials were conducted with mandarin and citron, two trials each for citrange, sour orange and sweet orange, and four trials for C. macrophylla. In citron the proportion of colonized leaves increased significantly over time, with 67, 85 and 96% of leaves colonized at 30, 60 and 90 days, respectively. For the other citrus types the exact proportion of colonized leaves differed, but colonization exceeded 60% in mandarin, sour orange, and citrange, and exceeded 80% at 30 days in two trials with sweet orange and three trials with C. macrophylla, but there was no significant increase in the proportion of colonized leaves at 60 and 90 days. Bacteria were readily detected by 30 days in new leaves of all citrus types. Differences in the growth of the bacterial population between citrus types and at different times of the year were noted, but common trends were apparent. In general, bacterial titers peaked at 60 days, except in leaves of C. macrophylla where bacterial titers peaked at 30 days. The early and consistently high proportion of leaf colonization observed for new growth of sweet orange during two trials and for C. macrophylla during three trials indicates a near synchronous colonization of new leaves by 30 days.

Citrus Research Board-sponsored review of the University of California Riverside citrus breeding

USDA-ARS?s Scientific Manuscript database

In October 2015 the Citrus Research Board (CRB) assembled a panel of experts to review the Citrus Research Board-sponsored Citrus Research and Genetics Programs at University of California Riverside (UCR). The panel consisted of: Gennaro Fazio, USDA/ARS, Geneva, NY; Maria Angeles Forner-Giner, Insti...

Combining 'omics and microscopy to visualize interactions between the Asian citrus psyllid vector and the Huanglongbing pathogen Candidatus Liberibacter asiaticus in the insect gut

USDA-ARS?s Scientific Manuscript database

Huanglongbing, or citrus greening disease, is an economically devastating bacterial disease of citrus. It is associated with infection by the gram-negative bacterium Candidatus Liberibacter asiaticus (CLas). CLas is transmitted by Diaphorina citri, the Asian citrus psyllid (ACP). For insect transmis...

A survey of FLS2 genes from multiple citrus species identifies candidates for enhancing disease resistance to Xanthomonas citri ssp. citri.

PubMed Central

Shi, Qingchun; Febres, Vicente J; Jones, Jeffrey B; Moore, Gloria A

2016-01-01

Pathogen-associated molecular patterns (PAMPs)-triggered immunity (PTI) is an important component of plant innate immunity. In a previous study, we showed that the PAMP flg22 from Xanthomonas citri ssp. citri (Xflg22), the causal agent of citrus canker, induced PTI in citrus, which correlated with the observed levels of canker resistance. Here, we identified and sequenced two bacterial flagellin/flg22 receptors (FLS2-1 and FLS2-2) from ‘Duncan’ grapefruit (Citrus paradisi, CpFLS2-1 and CpFLS2-2) and ‘Sun Chu Sha’ mandarin (C. reticulata, CrFLS2-1 and CrFLS2-2). We were able to isolate only one FLS2 from ‘Nagami’ kumquat (Fortunella margarita, FmFLS2-1) and gene flanking sequences suggest a rearrangement event that resulted in the deletion of FLS2-2 from the genome. Phylogenetic analysis, gene structure and presence of critical amino acid domains all indicate we identified the true FLS2 genes in citrus. FLS2-2 was more transcriptionally responsive to Xflg22 than FLS2-1, with induced expression levels higher in canker-resistant citrus than in susceptible ones. Interestingly, ‘Nagami’ kumquat showed the highest FLS2-1 steady-state expression levels, although it was not induced by Xflg22. We selected FmFLS2-1, CrFLS2-2 and CpFLS2-2 to further evaluate their capacity to enhance bacterial resistance using Agrobacterium-mediated transient expression assays. Both FmFLS2-1 and CrFLS2-2, the two proteins from canker-resistant species, conferred stronger Xflg22 responses and reduced canker symptoms in leaves of the susceptible grapefruit genotype. These two citrus genes will be useful resources to enhance PTI and achieve resistance against canker and possibly other bacterial pathogens in susceptible citrus types. PMID:27222722

Calculation of diagnostic parameters of advanced serological and molecular tissue-print methods for detection of Citrus tristeza virus. A model for other plant pathogens

USDA-ARS?s Scientific Manuscript database

Citrus tristeza virus (CTV) is one of the most important virus diseases which affect citrus. Control of CTV in Spain and central California is achieved by planting virus-free citrus on CTV-tolerant or -resistant rootstocks. Quarantine and certification programs remain essential to avoid importation ...

Arabidopsis Elongator subunit 2 positively contributes to resistance to the necrotrophic fungal pathogens Botrytis cinerea and Alternaria brassicicola.

PubMed

Wang, Chenggang; Ding, Yezhang; Yao, Jin; Zhang, Yanping; Sun, Yijun; Colee, James; Mou, Zhonglin

2015-09-01

The evolutionarily conserved Elongator complex functions in diverse biological processes including salicylic acid-mediated immune response. However, how Elongator functions in jasmonic acid (JA)/ethylene (ET)-mediated defense is unknown. Here, we show that Elongator is required for full induction of the JA/ET defense pathway marker gene PLANT DEFENSIN1.2 (PDF1.2) and for resistance to the necrotrophic fungal pathogens Botrytis cinerea and Alternaria brassicicola. A loss-of-function mutation in the Arabidopsis Elongator subunit 2 (ELP2) alters B. cinerea-induced transcriptome reprogramming. Interestingly, in elp2, expression of WRKY33, OCTADECANOID-RESPONSIVE ARABIDOPSIS AP2/ERF59 (ORA59), and PDF1.2 is inhibited, whereas transcription of MYC2 and its target genes is enhanced. However, overexpression of WRKY33 or ORA59 and mutation of MYC2 fail to restore PDF1.2 expression and B. cinerea resistance in elp2, suggesting that ELP2 is required for induction of not only WRKY33 and ORA59 but also PDF1.2. Moreover, elp2 is as susceptible as coronatine-insensitive1 (coi1) and ethylene-insensitive2 (ein2) to B. cinerea, indicating that ELP2 is an important player in B. cinerea resistance. Further analysis of the lesion sizes on the double mutants elp2 coi1 and elp2 ein2 and the corresponding single mutants revealed that the function of ELP2 overlaps with COI1 and is additive to EIN2 for B. cinerea resistance. Finally, basal histone acetylation levels in the coding regions of WRKY33, ORA59, and PDF1.2 are reduced in elp2 and a functional ELP2-GFP fusion protein binds to the chromatin of these genes, suggesting that constitutive ELP2-mediated histone acetylation may be required for full activation of the WRKY33/ORA59/PDF1.2 transcriptional cascade. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

Citrus and Prunuscopia-like retrotransposons.

PubMed

Asíns, M J; Monforte, A J; Mestre, P F; Carbonell, E A

1999-08-01

Many of the world's most important citrus cultivars ("Washington Navel", satsumas, clementines) have arisen through somatic mutation. This phenomenon occurs fairly often in the various species and varieties of the genus.The presence of copia-like retrotransposons has been investigated in fruit trees, especially citrus, by using a PCR assay designed to detect copia-like reverse transcriptase (RT) sequences. Amplification products from a genotype of each the following species Citrus sinensis, Citrus grandis, Citrus clementina, Prunus armeniaca and Prunus amygdalus, were cloned and some of them sequenced. Southern-blot hybridization using RT clones as probes showed that multiple copies are integrated throughout the citrus genome, while only 1-3 copies are detected in the P. armeniaca genome, which is in accordance with the Citrus and Prunus genome sizes. Sequence analysis of RT clones allowed a search for homologous sequences within three gene banks. The most similar ones correspond to RT domains of copia-like retrotransposons from unrelated plant species. Cluster analysis of these sequences has shown a great heterogeneity among RT domains cloned from the same genotype. This finding supports the hypothesis that horizontal transmission of retrotransposons has occurred in the past. The species presenting a RT sequence most similar to citrus RT clones is Gnetum montanum, a gymnosperm whose distribution area coincides with two of the main centers of origin of Citrus spp. A new C-methylated restriction DNA fragment containing a RT sequence is present in navel sweet oranges, but not in Valencia oranges from which the former originated suggesting, that retrotransposon activity might be, at least in part, involved in the genetic variability among sweet orange cultivars. Given that retrotransposons are quite abundant throughout the citrus genome, their activity should be investigated thoroughly before commercializing any transgenic citrus plant where the transgene(s) is part

Genome editing of the disease susceptibility gene CsLOB1 in citrus confers resistance to citrus canker.

PubMed

Jia, Hongge; Zhang, Yunzeng; Orbović, Vladimir; Xu, Jin; White, Frank F; Jones, Jeffrey B; Wang, Nian

2017-07-01

Citrus is a highly valued tree crop worldwide, while, at the same time, citrus production faces many biotic challenges, including bacterial canker and Huanglongbing (HLB). Breeding for disease-resistant varieties is the most efficient and sustainable approach to control plant diseases. Traditional breeding of citrus varieties is challenging due to multiple limitations, including polyploidy, polyembryony, extended juvenility and long crossing cycles. Targeted genome editing technology has the potential to shorten varietal development for some traits, including disease resistance. Here, we used CRISPR/Cas9/sgRNA technology to modify the canker susceptibility gene CsLOB1 in Duncan grapefruit. Six independent lines, D LOB 2, D LOB 3, D LOB 9, D LOB 10, D LOB 11 and D LOB 12, were generated. Targeted next-generation sequencing of the six lines showed the mutation rate was 31.58%, 23.80%, 89.36%, 88.79%, 46.91% and 51.12% for D LOB 2, D LOB 3, D LOB 9, D LOB 10, D LOB 11 and D LOB 12, respectively, of the cells in each line. D LOB 2 and D LOB 3 showed canker symptoms similar to wild-type grapefruit, when inoculated with the pathogen Xanthomonas citri subsp. citri (Xcc). No canker symptoms were observed on D LOB 9, D LOB 10, D LOB 11 and D LOB 12 at 4 days postinoculation (DPI) with Xcc. Pustules caused by Xcc were observed on D LOB 9, D LOB 10, D LOB 11 and D LOB 12 in later stages, which were much reduced compared to that on wild-type grapefruit. The pustules on D LOB 9 and D LOB 10 did not develop into typical canker symptoms. No side effects and off-target mutations were detected in the mutated plants. This study indicates that genome editing using CRISPR technology will provide a promising pathway to generate disease-resistant citrus varieties. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

One Target, Two Mechanisms: The Impact of 'Candidatus Liberibacter asiaticus' and Its Vector, Diaphorina citri, on Citrus Leaf Pigments.

PubMed

Killiny, Nabil; Nehela, Yasser

2017-07-01

Huanglongbing (HLB) is currently the largest threat to global citrus production. We examined the effect of HLB pathogen 'Candidatus Liberibacter asiaticus' infection or infestation by its vector, Diaphorina citri, on 'Valencia' sweet orange leaf pigments using high-performance liquid chromatography, followed by gene expression analysis for 46 involved genes in carotenoid and chlorophyll biosynthesis pathways. Both 'Ca. L. asiaticus' and D. citri alter the total citrus leaf pigment balance with a greater impact by 'Ca. L. asiaticus'. Although zeaxanthin was accumulated in 'Ca. L. asiaticus'-infected leaves, chlorophyllide a was increased in D. citri-infested plants. Our findings support the idea that both 'Ca. L. asiaticus' and D. citri affect the citrus pigments and promote symptom development but using two different mechanisms. 'Ca. L. asiaticus' promotes chlorophyll degradation but accelerates the biosynthesis of carotenoid pigments, resulting in accumulation of abscisic acid and its precursor, zeaxanthin. Zeaxanthin also has a photoprotective role. By contrast, D. citri induced the degradation of most carotenoids and accelerated chlorophyll biosynthesis, leading to chlorophyllide a accumulation. Chlorophyllide a might have an antiherbivory role. Accordingly, we suggest that citrus plants try to defend themselves against 'Ca. L. asiaticus' or D. citri using multifaceted defense systems, based on the stressor type. These findings will help in better understanding the tritrophic interactions among plant, pathogen, and vector.

Expressed Genes in Asian Citrus Psyllid adults feeding on citrus

USDA-ARS?s Scientific Manuscript database

We created and described the first genetic data set from the Asian citrus psyllid, AsCP, Diaphorina citri, Kuwayama (Hemiptera: Psyllidae). The AsCP spread the plant-infecting bacterium, Candidatus Liberibacter asiaticus, which is associated with the citrus disease Huanglongbing, HLB, known as Citru...

Two previously unknown Phytophthora species associated with brown rot of Pomelo (Citrus grandis) fruits in Vietnam.

PubMed

Puglisi, Ivana; De Patrizio, Alessandro; Schena, Leonardo; Jung, Thomas; Evoli, Maria; Pane, Antonella; Van Hoa, Nguyen; Van Tri, Mai; Wright, Sandra; Ramstedt, Mauritz; Olsson, Christer; Faedda, Roberto; Magnano di San Lio, Gaetano; Cacciola, Santa Olga

2017-01-01

Two distinct Phytophthora taxa were found to be associated with brown rot of pomelo (Citrus grandis), a new disease of this ancestral Citrus species, in the Vinh Long province, Mekong River Delta area, southern Vietnam. On the basis of morphological characters and using the ITS1-5.8S-ITS2 region of the rDNA and the cytochrome oxidase subunit 1 (COI) as barcode genes, one of the two taxa was provisionally named as Phytophthora sp. prodigiosa, being closely related to but distinct from P. insolita, a species in Phytophthora Clade 9, while the other one, was closely related to but distinct from the Clade 2 species P. meadii and was informally designated as Phytophthora sp. mekongensis. Isolates of P. sp. prodigiosa and P. sp. mekongensis were also obtained from necrotic fibrous roots of Volkamer lemon (C. volkameriana) rootstocks grafted with 'King' mandarin (Citrus nobilis) and from trees of pomelo, respectively, in other provinces of the Mekong River Delta, indicating a widespread occurrence of both Phytophthora species in this citrus-growing area. Koch's postulates were fulfilled via pathogenicity tests on fruits of various Citrus species, including pomelo, grapefruit (Citrus x paradisi), sweet orange (Citrus x sinensis) and bergamot (Citrus x bergamia) as well as on the rootstock of 2-year-old trees of pomelo and sweet orange on 'Carrizo' citrange (C. sinensis 'Washington Navel' x Poncirus trifoliata). This is the first report of a Phytophthora species from Clade 2 other than P. citricola and P. citrophthora as causal agent of fruit brown rot of Citrus worldwide and the first report of P. insolita complex in Vietnam. Results indicate that likely Vietnam is still an unexplored reservoir of Phytophthora diversity.

Two previously unknown Phytophthora species associated with brown rot of Pomelo (Citrus grandis) fruits in Vietnam

PubMed Central

Schena, Leonardo; Jung, Thomas; Evoli, Maria; Pane, Antonella; Van Hoa, Nguyen; Van Tri, Mai; Wright, Sandra; Ramstedt, Mauritz; Olsson, Christer; Faedda, Roberto; Magnano di San Lio, Gaetano

2017-01-01

Two distinct Phytophthora taxa were found to be associated with brown rot of pomelo (Citrus grandis), a new disease of this ancestral Citrus species, in the Vinh Long province, Mekong River Delta area, southern Vietnam. On the basis of morphological characters and using the ITS1-5.8S-ITS2 region of the rDNA and the cytochrome oxidase subunit 1 (COI) as barcode genes, one of the two taxa was provisionally named as Phytophthora sp. prodigiosa, being closely related to but distinct from P. insolita, a species in Phytophthora Clade 9, while the other one, was closely related to but distinct from the Clade 2 species P. meadii and was informally designated as Phytophthora sp. mekongensis. Isolates of P. sp. prodigiosa and P. sp. mekongensis were also obtained from necrotic fibrous roots of Volkamer lemon (C. volkameriana) rootstocks grafted with ‘King’ mandarin (Citrus nobilis) and from trees of pomelo, respectively, in other provinces of the Mekong River Delta, indicating a widespread occurrence of both Phytophthora species in this citrus-growing area. Koch’s postulates were fulfilled via pathogenicity tests on fruits of various Citrus species, including pomelo, grapefruit (Citrus x paradisi), sweet orange (Citrus x sinensis) and bergamot (Citrus x bergamia) as well as on the rootstock of 2-year-old trees of pomelo and sweet orange on ‘Carrizo’ citrange (C. sinensis ‘Washington Navel’ x Poncirus trifoliata). This is the first report of a Phytophthora species from Clade 2 other than P. citricola and P. citrophthora as causal agent of fruit brown rot of Citrus worldwide and the first report of P. insolita complex in Vietnam. Results indicate that likely Vietnam is still an unexplored reservoir of Phytophthora diversity. PMID:28208159

Inducible expression of Bs2 R gene from Capsicum chacoense in sweet orange (Citrus sinensis L. Osbeck) confers enhanced resistance to citrus canker disease.

PubMed

Sendín, Lorena Noelia; Orce, Ingrid Georgina; Gómez, Rocío Liliana; Enrique, Ramón; Grellet Bournonville, Carlos Froilán; Noguera, Aldo Sergio; Vojnov, Adrián Alberto; Marano, María Rosa; Castagnaro, Atilio Pedro; Filippone, María Paula

2017-04-01

Transgenic expression of the pepper Bs2 gene confers resistance to Xanthomonas campestris pv. vesicatoria (Xcv) pathogenic strains which contain the avrBs2 avirulence gene in susceptible pepper and tomato varieties. The avrBs2 gene is highly conserved among members of the Xanthomonas genus, and the avrBs2 of Xcv shares 96% homology with the avrBs2 of Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker disease. A previous study showed that the transient expression of pepper Bs2 in lemon leaves reduced canker formation and induced plant defence mechanisms. In this work, the effect of the stable expression of Bs2 gene on citrus canker resistance was evaluated in transgenic plants of Citrus sinensis cv. Pineapple. Interestingly, Agrobacterium-mediated transformation of epicotyls was unsuccessful when a constitutive promoter (2× CaMV 35S) was used in the plasmid construction, but seven transgenic lines were obtained with a genetic construction harbouring Bs2 under the control of a pathogen-inducible promoter, from glutathione S-transferase gene from potato. A reduction of disease symptoms of up to 70% was observed in transgenic lines expressing Bs2 with respect to non-transformed control plants. This reduction was directly dependent on the Xcc avrBs2 gene since no effect was observed when a mutant strain of Xcc with a disruption in avrBs2 gene was used for inoculations. Additionally, a canker symptom reduction was correlated with levels of the Bs2 expression in transgenic plants, as assessed by real-time qPCR, and accompanied by the production of reactive oxygen species. These results indicate that the pepper Bs2 resistance gene is also functional in a family other than the Solanaceae, and could be considered for canker control.

Citrus Huanglongbing tolerance in Australian Citrus Relatives, Microcitrus and Eremocirus

USDA-ARS?s Scientific Manuscript database

Tolerance, or resistance to citrus huanglongbing will be important as a long term solution for this disease. In a field trial conducted with over 1000 plants belonging to different genera in the sub-family Aurantioideae, we observed field tolerance in many Australian citrus relatives. To confirm the...

Phytochemical, antimicrobial, and antioxidant activities of different citrus juice concentrates.

PubMed

Oikeh, Ehigbai I; Omoregie, Ehimwenma S; Oviasogie, Faith E; Oriakhi, Kelly

2016-01-01

The search for new antimicrobial compounds is ongoing. Its importance cannot be overemphasized in an era of emerging resistant pathogenic organisms. This study therefore investigated the phytochemical composition and antioxidant and antimicrobial activities of different citrus juice concentrates. Fruit juices of Citrus tangerine (tangerine), Citrus paradisi (grape), Citrus limon (lemon), and Citrus aurantifolia (lime) were evaluated. Antimicrobial activities against five bacterial and three fungal strains were evaluated. The results revealed the presence of alkaloids, flavonoids, steroids, terpenoids, saponins, cardiac glycosides, and reducing sugars in all the juice concentrates. DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging capacities varied with tangerine and grape juices having better scavenging capacities than lemon and lime juices. Grape juice was observed to have a significantly higher (P < 0.05) ferric-reducing antioxidant potential (FRAP) value (364.2 ± 10.25 μmol/L Fe(II)/g of the extract) than the reference antioxidant, ascorbic acid (312.88 ± 5.61 μmol/L). Antimicrobial studies revealed differential antimicrobial activities against different microbial strains. Zones of inhibition ranging from 4 to 26 mm were observed for the antibacterial tests with 0-24 mm for antifungal test. Minimum inhibitory concentrations (MIC) and minimum bacteriostatic concentrations (MBC) for concentrates against bacterial strains ranged from 12.5 to 200 μg/mL. Lemon and lime juice concentrates had lower MIC and MBC values with orange and tangerine having the highest values. Minimum fungicidal concentrations ranged from 50 to 200 μg/mL. The results of this study suggest that these juice concentrates may have beneficial antimicrobial roles that can be exploited in controlling unwanted microbial growth.

Molecular characterization and phylogenetic analysis of Citrus viroid VI variants from citrus in China

USDA-ARS?s Scientific Manuscript database

Citrus viroid VI (CVd-VI) was originally found from citrus and persimmon in Japan. We report here the identification and molecular characterization of CVd-VI from four production regions of China. A total of 90 cDNA clones from nine infected citrus cultivars were sequenced. The sequence homologies o...

Annotation of the Asian Citrus Psyllid Genome Reveals a Reduced Innate Immune System

PubMed Central

Arp, Alex P.; Hunter, Wayne B.; Pelz-Stelinski, Kirsten S.

2016-01-01

Citrus production worldwide is currently facing significant losses due to citrus greening disease, also known as Huanglongbing. The citrus greening bacteria, Candidatus Liberibacter asiaticus (CLas), is a persistent propagative pathogen transmitted by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). Hemipterans characterized to date lack a number of insect immune genes, including those associated with the Imd pathway targeting Gram-negative bacteria. The D. citri draft genome was used to characterize the immune defense genes present in D. citri. Predicted mRNAs identified by screening the published D. citri annotated draft genome were manually searched using a custom database of immune genes from previously annotated insect genomes. Toll and JAK/STAT pathways, general defense genes Dual oxidase, Nitric oxide synthase, prophenoloxidase, and cellular immune defense genes were present in D. citri. In contrast, D. citri lacked genes for the Imd pathway, most antimicrobial peptides, 1,3-β-glucan recognition proteins (GNBPs), and complete peptidoglycan recognition proteins. These data suggest that D. citri has a reduced immune capability similar to that observed in A. pisum, P. humanus, and R. prolixus. The absence of immune system genes from the D. citri genome may facilitate CLas infections, and is possibly compensated for by their relationship with their microbial endosymbionts. PMID:27965582

Disruption of Vector Host Preference with Plant Volatiles May Reduce Spread of Insect-Transmitted Plant Pathogens.

PubMed

Martini, Xavier; Willett, Denis S; Kuhns, Emily H; Stelinski, Lukasz L

2016-05-01

Plant pathogens can manipulate the odor of their host; the odor of an infected plant is often attractive to the plant pathogen vector. It has been suggested that this odor-mediated manipulation attracts vectors and may contribute to spread of disease; however, this requires further broad demonstration among vector-pathogen systems. In addition, disruption of this indirect chemical communication between the pathogen and the vector has not been attempted. We present a model that demonstrates how a phytophathogen (Candidatus Liberibacter asiaticus) can increase its spread by indirectly manipulating the behavior of its vector (Asian citrus psyllid, Diaphorina citri Kuwayama). The model indicates that when vectors are attracted to pathogen-infected hosts, the proportion of infected vectors increases, as well as, the proportion of infected hosts. Additionally, the peak of infected host populations occurs earlier as compared with controls. These changes in disease dynamics were more important during scenarios with higher vector mortality. Subsequently, we conducted a series of experiments to disrupt the behavior of the Asian citrus psyllid. To do so, we exposed the vector to methyl salicylate, the major compound released following host infection with the pathogen. We observed that during exposure or after pre-exposure to methyl salicylate, the host preference can be altered; indeed, the Asian citrus psyllids were unable to select infected hosts over uninfected counterparts. We suggest mechanisms to explain these interactions and potential applications of disrupting herbivore host preference with plant volatiles for sustainable management of insect vectors.

Antioxidant Capacity, Anticancer Ability and Flavonoids Composition of 35 Citrus (Citrus reticulata Blanco) Varieties.

PubMed

Wang, Yue; Qian, Jing; Cao, Jinping; Wang, Dengliang; Liu, Chunrong; Yang, Rongxi; Li, Xian; Sun, Chongde

2017-07-05

Citrus ( Citrus reticulate Blanco) is one of the most commonly consumed and widely distributed fruit in the world, which is possessing extensive bioactivities. Present study aimed to fully understand the flavonoids compositions, antioxidant capacities and in vitro anticancer abilities of different citrus resources. Citrus fruits of 35 varieties belonging to 5 types (pummelos, oranges, tangerines, mandarins and hybrids) were collected. Combining li quid chromatography combined with electrospray ionization mass spectrometry (LC-ESI-MS/MS) and ultra-performance liquid chromatography combined with diode array detector (UPLC-DAD), a total of 39 flavonoid compounds were identified, including 4 flavones, 9 flavanones and 26 polymethoxylated flavonoids (PMFs). Each citrus fruit was examined and compared by 4 parts, flavedo, albedo, segment membrane and juice sacs. The juice sacs had the lowest total phenolics, following by the segment membrane. Four antioxidant traits including 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC) and cupric reducing antioxidant capacity (CUPRAC) were applied for the antioxidant capacities evaluation. Three gastric cancer cell lines, SGC-7901, BGC-823 and AGS were applied for the cytotoxicity evaluation. According to the results of correlation analysis, phenolics compounds might be the main contributor to the antioxidant activity of citrus extracts, while PMFs existing only in the flavedo might be closely related to the gastric cancer cell line cytotoxicity of citrus extracts. The results of present study might provide a theoretical guidance for the utilization of citrus resources.

First report of Phyllosticta citricarpa and description of two new species, P. paracapitalensis and P. paracitricarpa, from citrus in Europe.

PubMed

Guarnaccia, V; Groenewald, J Z; Li, H; Glienke, C; Carstens, E; Hattingh, V; Fourie, P H; Crous, P W

2017-06-01

The genus Phyllosticta occurs worldwide, and contains numerous plant pathogenic, endophytic and saprobic species. Phyllosticta citricarpa is the causal agent of Citrus Black Spot disease (CBS), affecting fruits and leaves of several citrus hosts ( Rutaceae ), and can also be isolated from asymptomatic citrus tissues. Citrus Black Spot occurs in citrus-growing regions with warm summer rainfall climates, but is absent in countries of the European Union (EU). Phyllosticta capitalensis is morphologically similar to P. citricarpa , but is a non-pathogenic endophyte, commonly isolated from citrus leaves and fruits and a wide range of other hosts, and is known to occur in Europe. To determine which Phyllosticta spp. occur within citrus growing regions of EU countries, several surveys were conducted (2015-2017) in the major citrus production areas of Greece, Italy, Malta, Portugal and Spain to collect both living plant material and leaf litter in commercial nurseries, orchards, gardens, backyards and plant collections. A total of 64 Phyllosticta isolates were obtained from citrus in Europe, of which 52 were included in a multi-locus (ITS, actA , tef1 , gapdh , LSU and rpb2 genes) DNA dataset. Two isolates from Florida (USA), three isolates from China, and several reference strains from Australia, South Africa and South America were included in the overall 99 isolate dataset. Based on the data obtained, two known species were identified, namely P. capitalensis (from asymptomatic living leaves of Citrus spp.) in Greece, Italy, Malta, Portugal and Spain, and P. citricarpa (from leaf litter of C . sinensis and C . limon ) in Italy, Malta and Portugal. Moreover, two new species were described, namely P. paracapitalensis (from asymptomatic living leaves of Citrus spp.) in Italy and Spain, and P. paracitricarpa (from leaf litter of C . limon ) in Greece. On a genotypic level, isolates of P . citricarpa populations from Italy and Malta (MAT1-2-1) represented a single clone

A Phagostimulant Blend for the Asian Citrus Psyllid.

PubMed

Lapointe, Stephen L; Hall, David G; George, Justin

2016-09-01

Chemical cues that elicit orientation by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), are of interest because it is the primary vector of the causal pathogen of citrus greening disease. Non-pesticidal control methods for D. citri remain a high priority for the citrus industry. While searching for semiochemicals that may be involved in orientation to host plants, we previously identified a blend of formic and acetic acids that stimulated substrate probing by D. citri. Here, we applied geometric mixture designs and response surface modeling to identify and optimize a 3-component blend that further increased the number of salivary sheaths produced by D. citri on a wax substrate containing a 3.5:1.6:1 blend of formic acid, acetic acid, and p-cymene, respectively. No evidence was found for remote orientation by D. citri adults through olfaction to the phagostimulant blends. Increased probing in response to the presence of phagostimulants in the wax matrix occurred after contact with the substrate. Yellow wax beads always attracted more D. citri adults and received more probes compared with white wax beads. Yellow beads containing the 3-component blend of phagostimulants were probed by D. citri 2 to 3 times more often compared with yellow beads alone. The phagostimulant effect also was tested by covering wax beads containing the 3-component blend with a plastic film to minimize olfaction or contact chemoreception by antennation. The plastic film did not affect the probing response, thus suggesting that chemosensation was associated with mouthparts and not olfactory receptors. Salivary sheaths produced in wax beads containing the phagostimulant blend were 4.5 times longer than sheaths produced in beads without tastants. This phenomenon might be used to improve a trap, design an attract-and-kill product, or enhance other means of managing D. citri and citrus greening disease.

The diversity of citrus endophytic bacteria and their interactions with Xylella fastidiosa and host plants.

PubMed

Azevedo, João Lúcio; Araújo, Welington Luiz; Lacava, Paulo Teixeira

2016-01-01

The bacterium Xylella fastidiosa is the causal agent of citrus variegated chlorosis (CVC) and has been associated with important losses in commercial orchards of all sweet orange [Citrus sinensis (L.)] cultivars. The development of this disease depends on the environmental conditions, including the endophytic microbial community associated with the host plant. Previous studies have shown that X. fastidiosa interacts with the endophytic community in xylem vessels as well as in the insect vector, resulting in a lower bacterial population and reduced CVC symptoms. The citrus endophytic bacterium Methylobacterium mesophilicum can trigger X. fastidiosa response in vitro, which results in reduced growth and induction of genes associated with energy production, stress, transport, and motility, indicating that X. fastidiosa has an adaptive response to M. mesophilicum. Although this response may result in reduced CVC symptoms, the colonization rate of the endophytic bacteria should be considered in studies that intend to use this endophyte to suppress CVC disease. Symbiotic control is a new strategy that uses symbiotic endophytes as biological control agents to antagonize or displace pathogens. Candidate endophytes for symbiotic control of CVC must occupy the xylem of host plants and attach to the precibarium of sharpshooter insects to access the pathogen. In the present review, we focus on interactions between endophytic bacteria from sweet orange plants and X. fastidiosa, especially those that may be candidates for control of CVC.

The diversity of citrus endophytic bacteria and their interactions with Xylella fastidiosa and host plants

PubMed Central

Azevedo, João Lúcio; Araújo, Welington Luiz; Lacava, Paulo Teixeira

2016-01-01

Abstract The bacterium Xylella fastidiosa is the causal agent of citrus variegated chlorosis (CVC) and has been associated with important losses in commercial orchards of all sweet orange [Citrus sinensis (L.)] cultivars. The development of this disease depends on the environmental conditions, including the endophytic microbial community associated with the host plant. Previous studies have shown that X. fastidiosa interacts with the endophytic community in xylem vessels as well as in the insect vector, resulting in a lower bacterial population and reduced CVC symptoms. The citrus endophytic bacterium Methylobacterium mesophilicum can trigger X. fastidiosa response in vitro, which results in reduced growth and induction of genes associated with energy production, stress, transport, and motility, indicating that X. fastidiosa has an adaptive response to M. mesophilicum. Although this response may result in reduced CVC symptoms, the colonization rate of the endophytic bacteria should be considered in studies that intend to use this endophyte to suppress CVC disease. Symbiotic control is a new strategy that uses symbiotic endophytes as biological control agents to antagonize or displace pathogens. Candidate endophytes for symbiotic control of CVC must occupy the xylem of host plants and attach to the precibarium of sharpshooter insects to access the pathogen. In the present review, we focus on interactions between endophytic bacteria from sweet orange plants and X. fastidiosa, especially those that may be candidates for control of CVC. PMID:27727362

Molecular mechanisms behind the accumulation of adenosine triphosphate (ATP) and H2O2 in citrus plants in response to ‘Candidatus Liberibacter asiaticus’ infection

USDA-ARS?s Scientific Manuscript database

Candidatus Liberibacter asiaticus (Las) is a fastidious, phloem-restricted pathogen with a significantly reduced genome, and attacks all citrus species with no immune cultivars documented to date. Like other plant bacterial pathogens, Las deploys effector proteins into the organelles of plant cells,...

Slight Fermentation with Lactobacillus fermentium Improves the Taste (Sugar:Acid Ratio) of Citrus (Citrus reticulata cv. chachiensis) Juice.

PubMed

Yu, Yuanshan; Xiao, Gengsheng; Xu, Yujuan; Wu, Jijun; Fu, Manqin; Wen, Jing

2015-11-01

The aim of this study was to evaluate the hypothesis that fermentation with Lactobacillus fermentium, which can metabolize citric acid, could be applied in improving the taste (sugar:acid ratio) of citrus juice. During fermentation, the strain of L. fermentium can preferentially utilize citric acid of citrus (Citrus reticulata cv. Chachiensis) juice to support the growth without the consumption of sugar. After 6 h of fermentation with L. fermentium at 30 °C, the sugar:acid ratio of citrus juice increased to 22:1 from 12:1, which resulted in that the hedonic scores of sweetness, acidity and overall acceptability of fermented-pasteurized citrus juice were higher than the unfermented-pasteurized citrus juice. Compared with unfermented-pasteurized citrus juice, the ORAC value and total amino acid showed a reduction, and no significant change (P > 0.05) in the L*, a*, b*, total soluble phenolics and ascorbic acid (Vc) content in the fermented-pasteurized citrus juice was observed as compared with unfermented-pasteurized citrus juice. Hence, slight fermentation with L. fermentium can be used for improving the taste (sugar:acid ratio) of citrus juice with the well retaining of quality. © 2015 Institute of Food Technologists®

An Ecoinformatics Approach to Field-Scale Evaluation of Insecticide Effects in California Citrus: Are Citrus Thrips and Citrus Red Mite Induced Pests?

PubMed

Livingston, George; Hack, Lindsey; Steinmann, Kimberly P; Grafton-Cardwell, Elizabeth E; Rosenheim, Jay A

2018-05-28

Experimental approaches to studying the consequences of pesticide use, including impacts on beneficial insects, are vital; however, they can be limited in scale and realism. We show that an ecoinformatics approach that leverages existing data on pesticides, pests, and beneficials across multiple fields can provide complementary insights. We do this using a multi-year dataset (2002-2013) on pesticide applications and density estimates of two pests, citrus thrips (Scirtothrips citri (Moulton [Thysanoptera: Thripidae])) and citrus red mites (Panonychus citri McGregor [Acari: Tetranychidae]), and a natural enemy (Euseius spp. predatory mites) collected from citrus groves in the San Joaquin Valley of California. Using correlative analyses, we investigated the long-term consequences of pesticide use on S. citri and P. citri population densities to evaluate the hypothesis that the pest status of these species is largely due to the disruption of natural biological control-i.e., these are induced pests. We also evaluated short-term pesticide efficacy (suppression of citrus thrips and citrus red mite populations immediately post-application) and asked if it was correlated with the suppression of Euseius predator populations. Although the short-term efficacy of different pesticides varied significantly, our dataset does not suggest that the use of citrus pesticides suppressed Euseius densities or worsened pest problems. We also find that there is no general trade-off between pesticide efficacy and pesticide risk to Eusieus, such that highly effective and minimally disruptive compounds were available to citrus growers during the studied time period.

Temporal Occurrence and Niche Preferences of Phytophthora spp. Causing Brown Rot of Citrus in the Central Valley of California.

PubMed

Hao, Wei; Miles, Timothy D; Martin, Frank N; Browne, Gregory T; Förster, Helga; Adaskaveg, James E

2018-03-01

Brown rot of citrus fruit is caused by several species of Phytophthora and is currently of serious concern for the California citrus industry. Two species, Phytophthora syringae and P. hibernalis, are quarantine pathogens in China, a major export market for California citrus. To maintain trade and estimate the risk of exporting a quarantine pathogen, the distribution and frequency of Phytophthora spp. causing brown rot of orange in major growing areas of California was investigated. Symptomatic fruit were collected from navel (winter to late spring) and Valencia (late spring to summer) orange orchards from 2013 to 2015. Species identification of isolates was based on morphological characteristics, random amplified polymorphic DNA banding patterns, and sequencing of the internal transcribed spacer and the partial cox2/spacer/cox1 regions from axenic cultures, or directly on DNA from fruit tissue using a multiplex TaqMan quantitative polymerase chain reaction assay. In winter samplings, the incidence of P. syringae based on the number of fruit with Phytophthora spp. detection ranged from 73.6 to 96.1% for the two counties surveyed. The remaining isolates were identified as P. citrophthora. In late spring or summer, only P. citrophthora was recovered. P. hibernalis and P. nicotianae were not detected in any fruit with brown rot symptoms. These results indicate that P. syringae is currently an important brown rot pathogen of citrus fruit in California during the cooler seasons of the year. In winter 2016 and 2017, P. syringae was recovered by pear baiting at a high incidence from leaf litter and from a small number of rhizosphere soil or root samples but not from living leaves on the tree. In contrast, P. citrophthora was rarely found in leaf litter but was commonly detected in the rhizosphere. Thus, leaf litter is a major inoculum source for P. syringae and this species occupies a distinct ecological niche.

Evaluation of resistance to asiatic citrus canker among selections of pera sweet orange (Citrus sinensis)

USDA-ARS?s Scientific Manuscript database

Asiatic citrus canker (ACC, caused by the bacterium Xanthomonas citri subsp. citri) is a destructive disease of citrus in Brazil and in several other citrus-producing countries. ACC management is problematic, and bactericides such as copper can be reasonably efficacious but do not completely control...

78 FR 8435 - Importation of Fresh Citrus Fruit From Uruguay, Including Citrus Hybrids and Fortunella

Federal Register 2010, 2011, 2012, 2013, 2014

2013-02-06

... gnidiella (the honeydew moth) and Gymnandrosoma aurantianum (citrus fruit borer); one fungus (Elsino[euml.... APHIS-2011-0060] RIN 0579-AD59 Importation of Fresh Citrus Fruit From Uruguay, Including Citrus Hybrids... Service, USDA. ACTION: Proposed rule. SUMMARY: We are proposing to amend the fruits and vegetables...

Repellent Activity of Botanical Oils against Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae).

PubMed

Kuhns, Emily H; Martini, Xavier; Hoyte, Angel; Stelinski, Lukasz L

2016-07-14

The Asian citrus psyllid, Diaphorina citri Kuwayama, is the insect vector of the pathogen causing huanglongbing. We selected three botanical oils to evaluate behavioral activity against D. citri. In laboratory olfactometer assays, fir oil was repellent to D. citri females, while litsea and citronella oils elicited no response from D. citri females. In choice settling experiments, D. citri settled almost completely on control plants rather than on plants treated with fir oil at a 9.5 mg/day release rate. Therefore, we conducted field trials to determine if fir oil reduced D. citri densities in citrus groves. We found no repellency of D. citri from sweet orange resets that were treated with fir oil dispensers releasing 10.4 g/day/tree as compared with control plots. However, we found a two-week decrease in populations of D. citri as compared with controls when the deployment rate of these dispensers was doubled. Our results suggest that treatment of citrus with fir oil may have limited activity as a stand-alone management tool for D. citri and would require integration with other management practices.

Citrus Waste Biomass Program

DOE Office of Scientific and Technical Information (OSTI.GOV)

Karel Grohman; Scott Stevenson

Renewable Spirits is developing an innovative pilot plant bio-refinery to establish the commercial viability of ehtanol production utilizing a processing waste from citrus juice production. A novel process based on enzymatic hydrolysis of citrus processing waste and fermentation of resulting sugars to ethanol by yeasts was successfully developed in collaboration with a CRADA partner, USDA/ARS Citrus and Subtropical Products Laboratory. The process was also successfully scaled up from laboratory scale to 10,000 gal fermentor level.

Spatial Genetic Structure of Coffee-Associated Xylella fastidiosa Populations Indicates that Cross Infection Does Not Occur with Sympatric Citrus Orchards.

PubMed

Francisco, Carolina S; Ceresini, Paulo C; Almeida, Rodrigo P P; Coletta-Filho, Helvécio D

2017-04-01

Xylella fastidiosa, an economically important plant-pathogenic bacterium, infects both coffee and citrus trees in Brazil. Although X. fastidiosa in citrus is well studied, knowledge about the population structure of this bacterium infecting coffee remains unknown. Here, we studied the population structure of X. fastidiosa infecting coffee trees in São Paulo State, Brazil, in four regions where citrus is also widely cultivated. Genotyping of over 500 isolates from coffee plants using 14 genomic microsatellite markers indicated that populations were largely geographically isolated, as previously found with populations of X. fastidiosa infecting citrus. These results were supported by a clustering analysis, which indicated three major genetic groups among the four sampled regions. Overall, approximately 38% of isolates showed significant membership coefficients not related to their original geographical populations (i.e., migrants), characterizing a significant degree of genotype flow among populations. To determine whether admixture occurred between isolates infecting citrus and coffee plants, one site with citrus and coffee orchards adjacent to each other was selected; over 100 isolates were typed from each host plant. No signal of natural admixture between citrus- and coffee-infecting isolates was found; artificial cross-infection assays with representative isolates also yielded no successful cross infection. A comparison determined that X. fastidiosa populations from coffee have higher genetic diversity and allelic richness compared with citrus. The results showed that coffee and citrus X. fastidiosa populations are effectively isolated from each other and, although coffee populations are spatially structured, migration has an important role in shaping diversity.

Alternaria and Cladosporium Fungal Allergen Epitopes are Denatured by Sodium Hypochlorite

PubMed Central

2009-01-01

Background Fungal allergens are ubiquitous; however, little progress has been made understanding fungal allergenic material removal from indoor environments. Purpose We investigated removal of environmental allergenic material derived from Alternaria and Cladosporium using sodium hypochlorite in vivo and in vitro. Methods Freeze dried allergen extract from Alternaria alternata and Cladosporium herbarum was treated with hypochlorite concentrations of 322, 88, 38, 16, 3, 0.3, and 0 mM, respectively. Remaining native allergenic material was quantified using enzyme immunoassay and remaining viable fungal material was evaluated. Results The results of treating Alternaria or Cladosporium extract with sodium hypochlorite are immediate and obvious. Concentrations greater than 100 mM remove color and concentrations between 100 and 38 mM partially uncolored the extract. Immunoassay for total antigenic and allergenic material remaining after treatment with sodium hypochlorite including 2 concentrations recommended for killing fungus confirmed a general destruction of antigenic and allergenic material at concentrations of 38 mM or greater. Conclusions This work confirms the ability of solutions of sodium hypochlorite to denature fungal allergenic material from common outdoor and indoor fungi A. alternata and C. herbarum. Destruction of recognized antigenic and allergenic epitopes occurs at hypochlorite concentrations commonly used for household cleaning. PMID:23282316

Structural and physiological analyses of the alkanesulphonate-binding protein (SsuA) of the citrus pathogen Xanthomonas citri.

PubMed

Tófoli de Araújo, Fabiano; Bolanos-Garcia, Victor M; Pereira, Cristiane T; Sanches, Mario; Oshiro, Elisa E; Ferreira, Rita C C; Chigardze, Dimitri Y; Barbosa, João Alexandre Gonçalves; de Souza Ferreira, Luís Carlos; Benedetti, Celso E; Blundell, Tom L; Balan, Andrea

2013-01-01

The uptake of sulphur-containing compounds plays a pivotal role in the physiology of bacteria that live in aerobic soils where organosulfur compounds such as sulphonates and sulphate esters represent more than 95% of the available sulphur. Until now, no information has been available on the uptake of sulphonates by bacterial plant pathogens, particularly those of the Xanthomonas genus, which encompasses several pathogenic species. In the present study, we characterised the alkanesulphonate uptake system (Ssu) of Xanthomonas axonopodis pv. citri 306 strain (X. citri), the etiological agent of citrus canker. A single operon-like gene cluster (ssuEDACB) that encodes both the sulphur uptake system and enzymes involved in desulphurisation was detected in the genomes of X. citri and of the closely related species. We characterised X. citri SsuA protein, a periplasmic alkanesulphonate-binding protein that, together with SsuC and SsuB, defines the alkanesulphonate uptake system. The crystal structure of SsuA bound to MOPS, MES and HEPES, which is herein described for the first time, provides evidence for the importance of a conserved dipole in sulphate group coordination, identifies specific amino acids interacting with the sulphate group and shows the presence of a rather large binding pocket that explains the rather wide range of molecules recognised by the protein. Isolation of an isogenic ssuA-knockout derivative of the X. citri 306 strain showed that disruption of alkanesulphonate uptake affects both xanthan gum production and generation of canker lesions in sweet orange leaves. The present study unravels unique structural and functional features of the X. citri SsuA protein and provides the first experimental evidence that an ABC uptake system affects the virulence of this phytopathogen.

Structural and Physiological Analyses of the Alkanesulphonate-Binding Protein (SsuA) of the Citrus Pathogen Xanthomonas citri

PubMed Central

Tófoli de Araújo, Fabiano; Bolanos-Garcia, Victor M.; Pereira, Cristiane T.; Sanches, Mario; Oshiro, Elisa E.; Ferreira, Rita C. C.; Chigardze, Dimitri Y.; Barbosa, João Alexandre Gonçalves; de Souza Ferreira, Luís Carlos; Benedetti, Celso E.; Blundell, Tom L.; Balan, Andrea

2013-01-01

Background The uptake of sulphur-containing compounds plays a pivotal role in the physiology of bacteria that live in aerobic soils where organosulfur compounds such as sulphonates and sulphate esters represent more than 95% of the available sulphur. Until now, no information has been available on the uptake of sulphonates by bacterial plant pathogens, particularly those of the Xanthomonas genus, which encompasses several pathogenic species. In the present study, we characterised the alkanesulphonate uptake system (Ssu) of Xanthomonas axonopodis pv. citri 306 strain (X. citri), the etiological agent of citrus canker. Methodology/Principal Findings A single operon-like gene cluster (ssuEDACB) that encodes both the sulphur uptake system and enzymes involved in desulphurisation was detected in the genomes of X. citri and of the closely related species. We characterised X. citri SsuA protein, a periplasmic alkanesulphonate-binding protein that, together with SsuC and SsuB, defines the alkanesulphonate uptake system. The crystal structure of SsuA bound to MOPS, MES and HEPES, which is herein described for the first time, provides evidence for the importance of a conserved dipole in sulphate group coordination, identifies specific amino acids interacting with the sulphate group and shows the presence of a rather large binding pocket that explains the rather wide range of molecules recognised by the protein. Isolation of an isogenic ssuA-knockout derivative of the X. citri 306 strain showed that disruption of alkanesulphonate uptake affects both xanthan gum production and generation of canker lesions in sweet orange leaves. Conclusions/Significance The present study unravels unique structural and functional features of the X. citri SsuA protein and provides the first experimental evidence that an ABC uptake system affects the virulence of this phytopathogen. PMID:24282519

Induced resistance against the Asian citrus psyllid, Diaphorina citri, by β-aminobutyric acid in citrus.

PubMed

Tiwari, Siddharth; Meyer, Wendy L; Stelinski, Lukasz L

2013-10-01

β-Aminobutyric acid (BABA) is known to induce resistance to microbial pathogens, nematodes and insects in several host plant/pest systems. The present study was undertaken to determine whether a similar effect of BABA occurred against the Asian citrus psyllid, Diaphorina citri Kuwayama, in citrus. A 25 mM drench application of BABA significantly reduced the number of eggs/plant as compared with a water control, whereas 200 and 100 mM applications of BABA reduced the numbers of nymphs/plant and adults/plants, respectively. A 5 mM foliar application of BABA significantly reduced the number of adults but not eggs or nymphs when compared with a water control treatment. In addition, leaf-dip bioassays using various concentrations (25–500 mM) of BABA indicated no direct toxic effect on 2nd and 5th instar nymphs or adult D. citri. BABA-treated plants were characterized by significantly lower levels of iron, magnesium, phosphorus, sodium, sulfur and zinc as compared with control plants. The expression level of the PR-2 gene (β-1,3-glucanase) in BABA-treated plants that were also damaged by D. citri adult feeding was significantly higher than in plants exposed to BABA, D. citri feeding alone or control plants. Our results indicate the potential for using BABA as a systemic acquired resistance management tool for D. citri.

Microjets of citrus fruit

NASA Astrophysics Data System (ADS)

Smith, Nicholas; Dickerson, Andrew

2017-11-01

The rupture of oil glands in the citrus exocarp is a common experience to the discerning citrus consumer. When peeled, oil cavities housed with the citrus exocarp often rupture outwardly in response to externally applied bending stresses. Bending of the peel compresses the soft material surrounding the glands, the albedo, increasing fluid pressure. Ultimately, the fluid pressure exceeds the failure strength of the outermost membrane, the flavedo. The ensuing high-velocity discharge of oil and exhaustive emptying of oil glands creates a novel method for jetting small quantities of the aromatic and volatile oil. We compare the jetting behavior across five citrus hybrids through high-speed videography and material testing of exocarps. The jetting oil undergoes an initial acceleration surpassing 5,000 gravities, reaching velocities in excess of 10 m/s. Film of citrus jets and mimicking jets in the lab reveal their high level of instability is caused by irregular and non-circular orifice geometry. Through material characterization and bending simulations, we rationalize the combination of material properties necessary to generate the internal gland pressures required for explosive dispersal.

Biocontrol ability and putative mode of action of yeasts against Geotrichum citri-aurantii in citrus fruit.

PubMed

Ferraz, Luriany Pompeo; Cunha, Tatiane da; da Silva, Aline Caroline; Kupper, Katia Cristina

2016-01-01

Sour rot is a major postharvest disease of citrus fruit and is caused by the fungal pathogen Geotrichum citri-aurantii. A lack of chemicals certified for the control of this disease has led to the consideration of alternative methods and strategies, such as the use of yeasts as biocontrol agents. The purpose of the present study was to test the ability of yeasts isolated from leaves, flowers, fruit, and soil, and six Saccharomyces cerevisiae isolates to control citrus sour rot, to assess the mechanisms of action of the yeast isolates that were demonstrated to be effective for biocontrol, and to identify the most effective yeast isolates for the biocontrol of G. citri-aurantii. In in vivo assays, three yeast isolates (ACBL-23, ACBL-44, and ACBL-77) showed a potential for controlling sour rot in citrus fruits, both preventatively and curatively. Most of the eight yeast isolates that were assessed for a mechanism of action did not produce antifungal compounds in an amount sufficient to inhibit the growth of the pathogen. Additionally, nutrient competition among the yeast strains was not found to be a biocontrol strategy. Instead, killer activity and hydrolytic enzyme production were identified as the major mechanisms involved in the biocontrol activity of the yeasts. Isolates ACBL-23, ACBL-44, and ACBL-77, which controlled sour rot most effectively, were identified as Rhodotorula minuta, Candida azyma, and Aureobasidium pullulans, respectively. To our knowledge, this is the first report of the potential of C. azyma as a biological control agent against a postharvest pathogen and its ability to produce a killer toxin. Copyright © 2016 Elsevier GmbH. All rights reserved.

Efficacy of Chaetomium Species as Biological Control Agents against Phytophthora nicotianae Root Rot in Citrus

PubMed Central

Wattanachai, Pongnak; Kasem, Soytong; Poeaim, Supattra

2015-01-01

Thailand is one of the largest citrus producers in Southeast Asia. Pathogenic infection by Phytophthora, however, has become one of major impediments to production. This study identified a pathogenic oomycete isolated from rotted roots of pomelo (Citrus maxima) in Thailand as Phytophthora nicotianae by the internal transcribed spacer ribosomal DNA sequence analysis. Then, we examined the in vitro and in vivo effects of Chaetomium globosum, Chaetomium lucknowense, Chaetomium cupreum and their crude extracts as biological control agents in controlling this P. nicotianae strain. Represent as antagonists in biculture test, the tested Chaetomium species inhibited mycelial growth by 50~56% and parasitized the hyphae, resulting in degradation of P. nicotianae mycelia after 30 days. The crude extracts of these Chaetomium species exhibited antifungal activities against mycelial growth of P. nicotianae, with effective doses of 2.6~101.4 µg/mL. Under greenhouse conditions, application of spores and methanol extracts of these Chaetomium species to pomelo seedlings inoculated with P. nicotianae reduced root rot by 66~71% and increased plant weight by 72~85% compared to that in the control. The method of application of antagonistic spores to control the disease was simple and economical, and it may thus be applicable for large-scale, highly effective biological control of this pathogen. PMID:26539045

Field Evaluation of Plant Defense Inducers for the Control of Citrus Huanglongbing.

PubMed

Li, Jinyun; Trivedi, Pankaj; Wang, Nian

2016-01-01

Huanglongbing (HLB) is currently the most economically devastating disease of citrus worldwide and no established cure is available. Defense inducing compounds are able to induce plant resistance effective against various pathogens. In this study the effects of various chemical inducers on HLB diseased citrus were evaluated in four groves (three with sweet orange and one with mandarin) in Florida (United States) for two to four consecutive growing seasons. Results have demonstrated that plant defense inducers including β-aminobutyric acid (BABA), 2,1,3-benzothiadiazole (BTH), and 2,6-dichloroisonicotinic acid (INA), individually or in combination, were effective in suppressing progress of HLB disease. Ascorbic acid (AA) and the nonmetabolizable glucose analog 2-deoxy-D-glucose (2-DDG) also exhibited positive control effects on HLB. After three or four applications for each season, the treatments AA (60 to 600 µM), BABA (0.2 to 1.0 mM), BTH (1.0 mM), INA (0.1 mM), 2-DDG (100 µM), BABA (1.0 mM) plus BTH (1.0 mM), BTH (1.0 mM) plus AA (600 µM), and BTH (1.0 mM) plus 2-DDG (100 µM) slowed down the population growth in planta of 'Candidatus Liberibacter asiaticus', the putative pathogen of HLB and reduced HLB disease severity by approximately 15 to 30% compared with the nontreated control, depending on the age and initial HLB severity of infected trees. These treatments also conferred positive effect on fruit yield and quality. Altogether, these findings indicate that plant defense inducers may be a useful strategy for the management of citrus HLB.

Colonization of citrus seed coats by 'Candidatus Liberibacter asiaticus': implications for seed transmission of the bacterium.

PubMed

Hilf, Mark E

2011-10-01

Huanglongbing is an economically damaging disease of citrus associated with infection by 'Candidatus Liberibacter asiaticus'. Transmission of the organism via infection of seeds has not been demonstrated but is a concern since some citrus varieties, particularly those used as rootstocks in commercial plantings are propagated from seed. We compared the incidence of detection of 'Ca. Liberibacter asiaticus' DNA in individual fruit peduncles, seed coats, seeds, and in germinated seedlings from 'Sanguenelli' sweet orange and 'Conners' grapefruit fruits sampled from infected trees. Using real-time quantitative PCR (qPCR) we detected pathogen DNA in nucleic acid extracts of 36 and 100% of peduncles from 'Sanguenelli' and from 'Conners' fruits, respectively. We also detected pathogen DNA in extracts of 37 and 98% of seed coats and in 1.6 and 4% of extracts from the corresponding seeds of 'Sanguenelli' and 'Conners', respectively. Small amounts of pathogen DNA were detected in 10% of 'Sanguenelli' seedlings grown in the greenhouse, but in none of 204 extracts from 'Conners' seedlings. Pathogen DNA was detected in 4.9% and in 89% of seed coats peeled from seeds of 'Sanguenelli' and 'Conners' which were germinated on agar, and in 5% of 'Sanguenelli' but in none of 164 'Conners' seedlings which grew from these seeds on agar. No pathogen DNA was detected in 'Ridge Pineapple' tissue at 3 months post-grafting onto 'Sanguenelli' seedlings, even when pathogen DNA had been detected initially in the 'Sanguenelli' seedling. Though the apparent colonization of 'Conners' seeds was more extensive and nearly uniform compared with 'Sanguenelli' seeds, no pathogen DNA was detected in 'Conners' seedlings grown from these seeds. For either variety, no association was established between the presence of pathogen DNA in fruit peduncles and seed coats and in seedlings.

Survey of Alternaria Toxins and Other Mycotoxins in Dried Fruits in China

PubMed Central

Wei, Dizhe; Wang, Yao; Jiang, Dongmei; Feng, Xiaoyuan; Li, Jun; Wang, Meng

2017-01-01

Occurrence of toxigenic molds and mycotoxins on dried fruits is a worldwide problem, but limited information is available in China. A total of 220 dried fruits (raisins, dried apricots, dates and wolfberries) purchased from China were analyzed for 17 mycotoxins (i.e., Alternaria toxins, ochratoxin A (OTA), patulin (PAT) and trichothecenes) by UPLC-MS/MS, combined with a single-step cleanup. The result showed that at least one mycotoxin was detected in 142 samples (64.6%). The lowest incidence of contaminated samples was observed in dried apricots (48.2%), and the highest incidence in dried wolfberries (83.3%). The Alternaria toxins seemed to be the major problem in dried fruits, rather than OTA or PAT. Tenuazonic acid (TeA) was the predominant mycotoxin, in both frequency and concentration, ranging from 6.9 to 5665.3 μg kg−1, followed by tentoxin (TEN; 20.5%), and mycophenolic acid (MPA; 19.5%). Moreover, raisins are more likely to be contaminated with OTA than the other dried fruits. Penicillic acid (PA) was detected only in dried dates, and PAT was detected only in one apricot sample. In addition, our results also showed that the simultaneous presence of 2–4 mycotoxins was observed in 31.4% of dried fruits. TeA and TEN were the most frequent combination, detected in 29 (13.2%) samples, followed by TeA and MPA with a prevalence of 11.4%. Therefore, the results of this survey suggest the need for wider monitoring on the contamination of these mycotoxins, especially Alternaria toxins in agro-products, and indicate the importance of setting a maximum limit for Alternaria toxins in China. PMID:28672847

Bacillus subtilis based-formulation for the control of postbloom fruit drop of citrus.

PubMed

Klein, Mariana Nadjara; da Silva, Aline Caroline; Kupper, Katia Cristina

2016-12-01

Postbloom fruit drop (PFD) caused by Colletotrichum acutatum affects flowers and causes early fruit drop in all commercial varieties of citrus. Biological control with the isolate ACB-69 of Bacillus subtilis has been considered as a potential method for controlling this disease. This study aimed to develop and optimize a B. subtilis based-formulation with a potential for large-scale applications and evaluate its effect on C. acutatum in vitro and in vivo. Bacillus subtilis based-formulations were developed using different carrier materials, and their ability to control PFD was evaluated. The results of the assays led to the selection of the B. subtilis based-formulation with talc + urea (0.02 %) and talc + ammonium molybdate (1 mM), which inhibited mycelial growth and germination of C. acutatum. Studies with detached citrus flowers showed that the formulations were effective in controlling the pathogen. In field conditions, talc + urea (0.02 %) provided 73 % asymptomatic citrus flowers and 56 % of the average number of effective fruit (ANEF), equating with fungicide treatment. On the contrary, non-treated trees had 8.8 % of asymptomatic citrus flowers and 0.83 % ANEF. The results suggest that B. subtilis based-formulations with talc as the carrier supplemented with a nitrogen source had a high potential for PFD control.

Rapid differentiation of citrus Hop stunt viroid variants by real-time RT-PCR and high resolution melting analysis.

PubMed

Loconsole, Giuliana; Onelge, Nuket; Yokomi, Raymond K; Kubaa, Raied Abou; Savino, Vito; Saponari, Maria

2013-01-01

The RNA genome of pathogenic and non-pathogenic variants of citrus Hop stunt viroid (HSVd) differ by five to six nucleotides located within the variable (V) domain referred to as the "cachexia expression motif". Sensitive hosts such as mandarin and its hybrids are seriously affected by cachexia disease. Current methods to differentiate HSVd variants rely on lengthy greenhouse biological indexing on Parson's Special mandarin and/or direct nucleotide sequence analysis of amplicons from RT-PCR of HSVd-infected plants. Two independent high throughput assays to segregate HSVd variants by real-time RT-PCR and High-Resolution Melting Temperature (HRM) analysis were developed: one based on EVAGreen dye; the other based on TaqMan probes. Primers for both assays targeted three differentiating nucleotides in the V domain which separated HSVd variants into three clusters by distinct melting temperatures with a confidence level higher than 98%. The accuracy of the HRM assays were validated by nucleotide sequencing of representative samples within each HRM cluster and by testing 45 HSVd-infected field trees from California, Italy, Spain, Syria and Turkey. To our knowledge, this is the first report of a rapid and sensitive approach to detect and differentiate HSVd variants associated with different biological behaviors. Although, HSVd is found in several crops including citrus, cachexia variants are restricted to some citrus-growing areas, particularly the Mediterranean Region. Rapid diagnosis for cachexia and non-cachexia variants is, thus, important for the management of HSVd in citrus and reduces the need for bioindexing and sequencing analysis. Copyright © 2013 Elsevier Ltd. All rights reserved.

First report of QoI resistance in Alternaria spp. infecting sugar beet (Beta vulgaris) in Michigan, USA

USDA-ARS?s Scientific Manuscript database

Alternaria leaf spot (ALS) of sugar beet is caused by Alternaria spp. in the A. alternata species complex. ALS is common wherever sugar beet is grown, but historically has been a minor issue for sugar beet production in the USA with damage usually not affecting crop yield significantly. Occurrence o...

Transcriptome analysis of root response to citrus blight based on the newly assembled Swingle citrumelo draft genome.

PubMed

Zhang, Yunzeng; Barthe, Gary; Grosser, Jude W; Wang, Nian

2016-07-08

Citrus blight is a citrus tree overall decline disease and causes serious losses in the citrus industry worldwide. Although it was described more than one hundred years ago, its causal agent remains unknown and its pathophysiology is not well determined, which hampers our understanding of the disease and design of suitable disease management. In this study, we sequenced and assembled the draft genome for Swingle citrumelo, one important citrus rootstock. The draft genome is approximately 280 Mb, which covers 74 % of the estimated Swingle citrumelo genome and the average coverage is around 15X. The draft genome of Swingle citrumelo enabled us to conduct transcriptome analysis of roots of blight and healthy Swingle citrumelo using RNA-seq. The RNA-seq was reliable as evidenced by the high consistence of RNA-seq analysis and quantitative reverse transcription PCR results (R(2) = 0.966). Comparison of the gene expression profiles between blight and healthy root samples revealed the molecular mechanism underneath the characteristic blight phenotypes including decline, starch accumulation, and drought stress. The JA and ET biosynthesis and signaling pathways showed decreased transcript abundance, whereas SA-mediated defense-related genes showed increased transcript abundance in blight trees, suggesting unclassified biotrophic pathogen was involved in this disease. Overall, the Swingle citrumelo draft genome generated in this study will advance our understanding of plant biology and contribute to the citrus breeding. Transcriptome analysis of blight and healthy trees deepened our understanding of the pathophysiology of citrus blight.

A Climatic Classification for Citrus Winter Survival in China.

NASA Astrophysics Data System (ADS)

Shou, Bo Huang

1991-05-01

The citrus tree is susceptible to frost damage. Winter injury to citrus from freezing weather is the major meteorological problem in the northern pail of citrus growing regions in China. Based on meteorological data collected at 120 stations in southern China and on the extent of citrus freezing injury, five climatic regions for citrus winter survival in China were developed. They were: 1) no citrus tree injury. 2) light injury to mandarins (citrus reticulate) or moderate injury to oranges (citrus sinensis), 3) moderate injury to mandarins or heavy injury to oranges, 4) heavy injury to mandarins, and 5) impossible citrus tree growth. This citrus climatic classification was an attempt to provide guidelines for regulation of citrus production, to effectively utilize land and climatic resources, to chose suitable citrus varieties, and to develop methods to prevent injury by freezing.

Citrus leprosis virus N: A New Dichorhavirus Causing Citrus Leprosis Disease.

PubMed

Ramos-González, Pedro Luis; Chabi-Jesus, Camila; Guerra-Peraza, Orlene; Tassi, Aline Daniele; Kitajima, Elliot Watanabe; Harakava, Ricardo; Salaroli, Renato Barbosa; Freitas-Astúa, Juliana

2017-08-01

Citrus leprosis (CL) is a viral disease endemic to the Western Hemisphere that produces local necrotic and chlorotic lesions on leaves, branches, and fruit and causes serious yield reduction in citrus orchards. Samples of sweet orange (Citrus × sinensis) trees showing CL symptoms were collected during a survey in noncommercial citrus areas in the southeast region of Brazil in 2013 to 2016. Transmission electron microscopy analyses of foliar lesions confirmed the presence of rod-like viral particles commonly associated with CL in the nucleus and cytoplasm of infected cells. However, every attempt to identify these particles by reverse-transcription polymerase chain reaction tests failed, even though all described primers for the detection of known CL-causing cileviruses and dichorhaviruses were used. Next-generation sequencing of total RNA extracts from three symptomatic samples revealed the genome of distinct, although highly related (>92% nucleotide sequence identity), viruses whose genetic organization is similar to that of dichorhaviruses. The genome sequence of these viruses showed <62% nucleotide sequence identity with those of orchid fleck virus and coffee ringspot virus. Globally, the deduced amino acid sequences of the open reading frames they encode share 32.7 to 63.8% identity with the proteins of the dichorhavirids. Mites collected from both the naturally infected citrus trees and those used for the transmission of one of the characterized isolates to Arabidopsis plants were anatomically recognized as Brevipalpus phoenicis sensu stricto. Molecular and biological features indicate that the identified viruses belong to a new species of CL-associated dichorhavirus, which we propose to call Citrus leprosis N dichorhavirus. Our results, while emphasizing the increasing diversity of viruses causing CL disease, lead to a reevaluation of the nomenclature of those viruses assigned to the genus Dichorhavirus. In this regard, a comprehensive discussion is

Effect of postharvest practices including degreening on citrus carpoplane microbial biomes.

PubMed

Gomba, A; Chidamba, L; Korsten, L

2017-04-01

To investigate the effect of commercial citrus packhouse processing steps on the fruit surface microbiome of Clementines and Palmer navel oranges. Viable bacteria, yeast and fungi counts, and the pyrosequencing analysis of the 16S rRNA and ITS were used to evaluate the community structure and population dynamics of phylloepiphytic bacteria and fungi associated with commercial postharvest processing. Drenching significantly reduced microbial counts in all cases except for yeasts on navels, while the extent of degreening effects varied between the citrus varieties. Pyrosequencing analysis showed a total of 4409 bacteria and 5792 fungi nonchimeric unique sequences with an average of 1102 bacteria and 1448 fungi reads per sample. Dominant phyla on the citrus carpoplane were Proteobacteria (53·5%), Actinobacteria (19·9%), Bacteroidetes (5·6%) and Deinococcus-Thermus (5·4%) for bacteria and Ascomycota (80·5%) and Basidiomycota (9·8%) for fungi. Beginning with freshly harvested fruit fungal diversity declined significantly after drenching, but had little effect on bacteria and populations recovered during degreening treatments, including those for Penicillium sp. Packhouse processing greatly influences microbial communities on the citrus carpoplane. A broad orange biome was described with pyrosequencing and gave insight into the likely survival and persistence of pathogens, especially as they may affect the quality and safety of the packed product. A close examination of the microbiota of fruit and the impact of intervention strategies on the ecological balance may provide a more durable approach to reduce losses and spoilage. © 2017 The Society for Applied Microbiology.

Identification of huanglongbing tolerance-associated genes using Candidatus Liberibacter asiaticus flagellin 22 as a proxy to challenge citrus

USDA-ARS?s Scientific Manuscript database

Plant defense elicited by pathogen-associated molecular patterns (PAMPs) is an important component of disease resistance. Previous research indicated the canker resistance in citrus correlates with responsiveness to Xcc-flg22, the 22 amino acid PAMP from the flagellin of Xanthomonas citri ssp. citri...

Alternaria leaf spot in Michigan and fungicide sensitivity issues

USDA-ARS?s Scientific Manuscript database

Since 2010 there has been an increase in identification of Alternaria leaf spot on sugar beet in Michigan and other growing regions in the US and Canada. In 2016, the disease was severe enough to cause economic losses in the Michigan growing region. Michigan isolates from sugar beet were examined ...

Ethyl p-coumarate exerts antifungal activity in vitro and in vivo against fruit Alternaria alternata via membrane-targeted mechanism.

PubMed

Li, Wusun; Yuan, Shuzhi; Sun, Jing; Li, Qianqian; Jiang, Weibo; Cao, Jiankang

2018-08-02

The fungus Alternaria alternata can cause food contamination by black spot rot and food safety issues due to the production of mycotoxins. In particular, A. alternata can infect many fresh fruits and vegetables and lead to considerable postharvest decay during storage and processing. The use of plant-derived products in postharvest disease management may be an acceptable alternative to traditional chemical fungicides. The aim of this study was to assess the antifungal activity of ethyl p-coumarate (EpCA) against Alternaria alternata in vitro and in vivo, and to determine the underlying mechanism. Results indicated that EpCA exhibited pronounced antifungal activity against in vitro mycelial growth of A. alternata, with half-inhibition concentration of 176.8 μg/mL. Spore germination of the pathogen was inhibited by EpCA in a dose-dependent manner. Moreover, in vivo test confirmed that both 100 and 800 μg/mL EpCA significantly reduced disease development of black spot rot in jujube fruit caused by A. alternata. The EpCA treatments increased plasma membrane permeability as great leakage of intercellular electrolytes, soluble proteins and sugars of A. alternata occurred during incubation. The EpCA treatments also caused increase of the influx of propidium iodide, a fluorescence dye binding nucleus DNA, into the affected spores, indicating the disrupted plasma membrane integrity. Observations of ultrastructure further evidenced the damage to plasma membrane and morphology of A. alternata caused by EpCA, which resulted in distortion, sunken and shrivelled of spores and mycelia of the pathogen. In addition, fluorometric assay by confocal laser scanning microscopy confirmed that the EpCA treatments induced endogenous reactive oxygen species (ROS) formation in the spores of A. alternata, with stronger and more stable accumulation of ROS at higher concentration of EpCA. Therefore, heavy oxidative damage to cellular membranes and organelles might happen as demonstrated

Diaphorina citri Induces Huanglongbing-Infected Citrus Plant Volatiles to Repel and Reduce the Performance of Propylaea japonica.

PubMed

Lin, Yongwen; Lin, Sheng; Akutse, Komivi S; Hussain, Mubasher; Wang, Liande

2016-01-01

Transmission of plant pathogens through insect vectors is a complex biological process involving interactions between the host plants, insects, and pathogens. Simultaneous impact of the insect damage and pathogenic bacteria in infected host plants induce volatiles that modify not only the behavior of its insect vector but also of their natural enemies, such as parasitoid wasps. Therefore, it is essential to understand how insects such as the predator ladybird beetle responds to volatiles emitted from a host plant and how the disease transmission alters the interactions between predators, vector, pathogens, and plants. In this study, we investigated the response of Propylaea japonica to volatiles from citrus plants damaged by Diaphorina citri and Candidatus Liberibacter asiaticus through olfactometer bioassays. Synthetic chemical blends were also used to determine the active compounds in the plant volatile. The results showed that volatiles emitted by healthy plants attracted more P. japonica than other treatments, due to the presence of high quantities of D-limonene and beta-ocimene, and the lack of methyl salicylate. When using synthetic chemicals in the olfactory tests, we found that D-limonene attracted P. japonica while methyl salicylate repelled the predator. However, beta-ocimene attracted the insects at lower concentrations but repelled them at higher concentrations. These results indicate that P. japonica could not efficiently search for its host by using volatile cues emitted from psyllids- and Las bacteria-infected citrus plants.

Deciphering the Bacterial Microbiome in Huanglongbing-Affected Citrus Treated with Thermotherapy and Sulfonamide Antibiotics

PubMed Central

Powell, Charles A.; Duan, Yongping; Shatters, Robert; Fang, Jingping; Zhang, Muqing

2016-01-01

Huanglongbing (HLB) is a serious citrus disease that threatens the citrus industry. In previous studies, sulfonamide antibiotics and heat treatment suppressed ‘Candidatus Liberibacter asiaticus’ (Las), but did not completely eliminate the Las. Furthermore, there are few reports studying the bacterial microbiome of HLB-affected citrus treated by heat and sulfonamide antibiotics. In this study, combinations of heat (45°C or 40°C) and sulfonamide treatment (sulfathiazole sodium–STZ, or sulfadimethoxine sodium—SDX) were applied to HLB-affected citrus. The bacterial microbiome of HLB-affected citrus following thermotherapy and/or chemotherapy was characterized by PhyloChipTMG3-based metagenomics. Our results showed that the combination of thermotherapy at 45°C and chemotherapy with STZ and SDX was more effective against HLB than thermotherapy alone, chemotherapy alone, or a combination of thermotherapy at 40°C and chemotherapy. The PhyloChipTMG3-based results indicated that 311 empirical Operational Taxonomic Units (eOTUs) were detected in 26 phyla. Cyanobacteria (18.01%) were dominant after thermo-chemotherapy. Thermotherapy at 45°C decreased eOTUs (64.43%) in leaf samples, compared with thermotherapy at 40°C (73.96%) or without thermotherapy (90.68%) and it also reduced bacterial family biodiversity. The eOTU in phylum Proteobacteria was reduced significantly and eOTU_28, representing “Candidatus Liberibacter,” was not detected following thermotherapy at 45°C. Following antibiotic treatment with SDX and STZ, there was enhanced abundance of specific eOTUs belonging to the families Streptomycetaceae, Desulfobacteraceae, Chitinophagaceae, and Xanthomonadaceae, which may be implicated in increased resistance to plant pathogens. Our study further develops an integrated strategy for combating HLB, and also provides new insight into the bacterial microbiome of HLB-affected citrus treated by heat and sulfonamide antibiotics. PMID:27171468

Terpene down-regulation in orange reveals the role of fruit aromas in mediating interactions with insect herbivores and pathogens.

PubMed

Rodríguez, Ana; San Andrés, Victoria; Cervera, Magdalena; Redondo, Ana; Alquézar, Berta; Shimada, Takehiko; Gadea, José; Rodrigo, María Jesús; Zacarías, Lorenzo; Palou, Lluís; López, María M; Castañera, Pedro; Peña, Leandro

2011-06-01

Plants use volatile terpene compounds as odor cues for communicating with the environment. Fleshy fruits are particularly rich in volatiles that deter herbivores and attract seed dispersal agents. We have investigated how terpenes in citrus fruit peels affect the interaction between the plant, insects, and microorganisms. Because limonene represents up to 97% of the total volatiles in orange (Citrus sinensis) fruit peel, we chose to down-regulate the expression of a limonene synthase gene in orange plants by introducing an antisense construct of this gene. Transgenic fruits showed reduced accumulation of limonene in the peel. When these fruits were challenged with either the fungus Penicillium digitatum or with the bacterium Xanthomonas citri subsp. citri, they showed marked resistance against these pathogens that were unable to infect the peel tissues. Moreover, males of the citrus pest medfly (Ceratitis capitata) were less attracted to low limonene-expressing fruits than to control fruits. These results indicate that limonene accumulation in the peel of citrus fruit appears to be involved in the successful trophic interaction between fruits, insects, and microorganisms. Terpene down-regulation might be a strategy to generate broad-spectrum resistance against pests and pathogens in fleshy fruits from economically important crops. In addition, terpene engineering may be important for studying the basic ecological interactions between fruits, herbivores, and pathogens.

Citrus Inventory

NASA Technical Reports Server (NTRS)

1994-01-01

An aerial color infrared (CIR) mapping system developed by Kennedy Space Center enables Florida's Charlotte County to accurately appraise its citrus groves while reducing appraisal costs. The technology was further advanced by development of a dual video system making it possible to simultaneously view images of the same area and detect changes. An image analysis system automatically surveys and photo interprets grove images as well as automatically counts trees and reports totals. The system, which saves both time and money, has potential beyond citrus grove valuation.

Citrus Inventory

NASA Technical Reports Server (NTRS)

1986-01-01

Florida's Charlotte County Property Appraiser is using an aerial color infrared mapping system for inventorying citrus trees for valuation purposes. The ACIR system has significantly reduced the time and manpower required for appraisal. Aerial photographs are taken and interpreted by a video system which makes it possible to detect changes from previous years. Potential problems can be identified. KSC's TU Office has awarded a contract to the Citrus Research and Education Center to adapt a prototype system which would automatically count trees and report totals.

Weeping dragon, a unique ornamenal citrus

USDA-ARS?s Scientific Manuscript database

‘Weeping Dragon’ is a new ornamental citrus cultivar developed by intercrossing of two unusual and unique citrus types, Poncirus trifoliata cultivated variety (cv.) Flying Dragon, and Citrus sinensis cv. ‘Cipo’. This new hybrid cultivar combines strongly contorted and weeping growth traits in a smal...

Isolation and identification of citrus psorosis virus Egyptian isolate (CPsV-EG).

PubMed

Ghazal, S A; El-Dougdoug, Kh A; Mousa, A A; Fahmy, H; Sofy, A R

2008-01-01

Citrus psorosis ophiovirus (CPsV), is considered to be of the most serious and deter mental virus pathogen's citrus species trees in Egypt. CPsV-EG was isolated from infected citrus grapefruit (C. paradisi Macf.) at Agric. Res. Centre (ARC). The grapefruit which used for CPsV-EG isolate was found to be free from CTV, CEVd and Spiroplasma citri where as gave -ve results with DTBIA, tissue print hybridization and Diene's stain respectively. CPsV-EG was detected on the basis of biological indexing by graft inoculation which gave oak leaf pattern (OLP) on Dweet tangor and serological assay by DAS-ELISA using Mab specific CPsV. CPsV-EG was reacted with variable responses on 16 host plants belonging to 6 families. Only 8 host plants are susceptible and showed visible external symptoms which appeared as local, systemic and local followed by systemic infections. CPsV-EG isolate was transmitted from infected citrus to citrus by syringe and grafting and herbaceous plants by forefinger inoculation and syringe. The woody indicators and rootstocks were differed in response to CPsV-EG isolate which appeared as no-response, response, sensitivity and hypersensitivity. The serological characters represented as the antigenic determinants of CPsV-EG isolate related to monoclonal antibodies specific CPsV strain where as appeared precipitation reaction by DAS-ELISA and DTBIA. The partial fragment of RNA3 (coat protein gene) of CPsV-EG (-1140bp and -571bp) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from grapefruit tissues using two sets primers specific CPsV (CPV3 and CPV4) and (PS66 and PS65) respectively. The virus under study was identified as CPsV-EG isolate according to biological, serological and molecular characters.

Antimicrobial nanoemulsion formulation with improved penetration of foliar spray through citrus leaf cuticles to control citrus Huanglongbing

USDA-ARS?s Scientific Manuscript database

Huanglongbing (HLB) is one of the most serious citrus diseases that threaten citrus industry worldwide. Because Candidatus Liberibacter asiaticus (Las) resides in citrus phloem, it is difficult to deliver an effective chemical compound into the phloem for control of HLB. In this study, a transcuticu...

78 FR 41259 - Importation of Fresh Citrus Fruit From Uruguay, Including Citrus

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-10

... net increase in the U.S. supply of fresh citrus varieties, as well as some displacement of the... statement in the economic analysis that ``any product displacement that may occur because of the proposed... share (product displacement) for current foreign suppliers of fresh citrus to the United States. U.S...

Repellent Activity of Botanical Oils against Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae)

PubMed Central

Kuhns, Emily H.; Martini, Xavier; Hoyte, Angel; Stelinski, Lukasz L.

2016-01-01

The Asian citrus psyllid, Diaphorina citri Kuwayama, is the insect vector of the pathogen causing huanglongbing. We selected three botanical oils to evaluate behavioral activity against D. citri. In laboratory olfactometer assays, fir oil was repellent to D. citri females, while litsea and citronella oils elicited no response from D. citri females. In choice settling experiments, D. citri settled almost completely on control plants rather than on plants treated with fir oil at a 9.5 mg/day release rate. Therefore, we conducted field trials to determine if fir oil reduced D. citri densities in citrus groves. We found no repellency of D. citri from sweet orange resets that were treated with fir oil dispensers releasing 10.4 g/day/tree as compared with control plots. However, we found a two-week decrease in populations of D. citri as compared with controls when the deployment rate of these dispensers was doubled. Our results suggest that treatment of citrus with fir oil may have limited activity as a stand-alone management tool for D. citri and would require integration with other management practices. PMID:27429006

Small RNA profiling reveals phosphorus deficiency as a contributing factor in symptom expression for citrus huanglongbing disease.

PubMed

Zhao, Hongwei; Sun, Ruobai; Albrecht, Ute; Padmanabhan, Chellappan; Wang, Airong; Coffey, Michael D; Girke, Thomas; Wang, Zonghua; Close, Timothy J; Roose, Mikeal; Yokomi, Raymond K; Folimonova, Svetlana; Vidalakis, Georgios; Rouse, Robert; Bowman, Kim D; Jin, Hailing

2013-03-01

Huanglongbing (HLB) is a devastating citrus disease that is associated with bacteria of the genus 'Candidatus Liberibacter' (Ca. L.). Powerful diagnostic tools and management strategies are desired to control HLB. Host small RNAs (sRNA) play a vital role in regulating host responses to pathogen infection and are used as early diagnostic markers for many human diseases, including cancers. To determine whether citrus sRNAs regulate host responses to HLB, sRNAs were profiled from Citrus sinensis 10 and 14 weeks post grafting with Ca. L. asiaticus (Las)-positive or healthy tissue. Ten new microRNAs (miRNAs), 76 conserved miRNAs, and many small interfering RNAs (siRNAs) were discovered. Several miRNAs and siRNAs were highly induced by Las infection, and can be potentially developed into early diagnosis markers of HLB. miR399, which is induced by phosphorus starvation in other plant species, was induced specifically by infection of Las but not Spiroplasma citri that causes citrus stubborn-a disease with symptoms similar to HLB. We found a 35% reduction of phosphorus in Las-positive citrus trees compared to healthy trees. Applying phosphorus oxyanion solutions to HLB-positive sweet orange trees reduced HLB symptom severity and significantly improved fruit production during a 3-year field trial in south-west Florida. Our molecular, physiological, and field data suggest that phosphorus deficiency is linked to HLB disease symptomology.

Green synthesis of gold nanoparticles using Citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) aqueous extract and its characterization.

PubMed

Sujitha, Mohanan V; Kannan, Soundarapandian

2013-02-01

This study reports the biological synthesis of gold nanoparticles by the reduction of HAuCl(4) by using citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) juice extract as the reducing and stabilizing agent. A various shape and size of gold nanoparticles were formed when the ratio of the reactants were altered with respect to 1.0mM chloroauric acid solution. The gold nanoparticles obtained were characterized by UV-visible spectra, transmission electron microscopy (TEM) and X-ray diffraction (XRD). TEM studies showed the particles to be of various shapes and sizes and particle size ranges from 15 to 80 nm. Selected-area electron diffraction (SAED) pattern confirmed fcc phase and crystallinity of the particles. The X-ray diffraction analysis revealed the distinctive facets (111, 200, 220 and 222 planes) of gold nanoparticles. Dynamic light scattering (DLS) studies revealed that the average size for colloid gp(3) of C. limon, C. reticulata and C. sinensis are 32.2 nm, 43.4 nm and 56.7 nm respectively. The DLS graph showed that the particles size was larger and more polydispersed compared to the one observed by TEM due to the fact that the measured size also includes the bio-organic compounds enveloping the core of the Au NPs. Zeta potential value for gold nanoparticles obtained from colloid gp(3) of C. limon, C. reticulata and C. sinensis are -45.9, -37.9 and -31.4 respectively indicating the stability of the synthesized nanoparticles. Herein we propose a novel, previously unexploited method for the biological syntheses of polymorphic gold nanoparticles with potent biological applications. Copyright © 2012 Elsevier B.V. All rights reserved.

Accumulation of transcription factors and cell signaling-related proteins in the nucleus during citrus-Xanthomonas interaction.

PubMed

Rani, T Swaroopa; Durgeshwar, P; Podile, Appa Rao

2015-07-20

The nucleus is the maestro of the cell and is involved in the modulation of cell signaling during stress. We performed a comprehensive nuclear proteome analysis of Citrus sinensis during interaction with host (Xanthomonas citri pv. citri-Xcc) and non-host (Xanthomonas oryzae pv. oryzae-Xoo) pathogens. The nuclear proteome was obtained using a sequential method of organelle enrichment and determined by nano-LC-MS/MS analysis. A total of 243 proteins accumulated differentially during citrus-Xanthomonas interaction, belonging to 11 functional groups, with signaling and transcription-related proteins dominating. MADS-box transcription factors, DEAD-box RNA helicase and leucine aminopeptidase, mainly involved in jasmonic acid (JA) responses, were in high abundance during non-host interaction (Xoo). Signaling-related proteins like serine/threonine kinase, histones (H3.2, H2A), phosphoglycerate kinase, dynamin, actin and aldolase showed increased accumulation early during Xoo interaction. Our results suggest that there is a possible involvement of JA-triggered defense responses during non-host resistance, with early recognition of the non-host pathogen. Copyright © 2015. Published by Elsevier GmbH.

The LOV Protein of Xanthomonas citri subsp. citri Plays a Significant Role in the Counteraction of Plant Immune Responses during Citrus Canker

PubMed Central

Kraiselburd, Ivana; Daurelio, Lucas D.; Tondo, María Laura; Merelo, Paz; Cortadi, Adriana A.; Talón, Manuel; Tadeo, Francisco R.; Orellano, Elena G.

2013-01-01

Pathogens interaction with a host plant starts a set of immune responses that result in complex changes in gene expression and plant physiology. Light is an important modulator of plant defense response and recent studies have evidenced the novel influence of this environmental stimulus in the virulence of several bacterial pathogens. Xanthomonas citri subsp. citri is the bacterium responsible for citrus canker disease, which affects most citrus cultivars. The ability of this bacterium to colonize host plants is influenced by bacterial blue-light sensing through a LOV-domain protein and disease symptoms are considerably altered upon deletion of this protein. In this work we aimed to unravel the role of this photoreceptor during the bacterial counteraction of plant immune responses leading to citrus canker development. We performed a transcriptomic analysis in Citrus sinensis leaves inoculated with the wild type X. citri subsp. citri and with a mutant strain lacking the LOV protein by a cDNA microarray and evaluated the differentially regulated genes corresponding to specific biological processes. A down-regulation of photosynthesis-related genes (together with a corresponding decrease in photosynthesis rates) was observed upon bacterial infection, this effect being more pronounced in plants infected with the lov-mutant bacterial strain. Infection with this strain was also accompanied with the up-regulation of several secondary metabolism- and defense response-related genes. Moreover, we found that relevant plant physiological alterations triggered by pathogen attack such as cell wall fortification and tissue disruption were amplified during the lov-mutant strain infection. These results suggest the participation of the LOV-domain protein from X. citri subsp. citri in the bacterial counteraction of host plant defense response, contributing in this way to disease development. PMID:24260514

Thymus vulgaris essential oil and thymol against Alternaria alternata (Fr.) Keissler: effects on growth, viability, early infection and cellular mode of action.

PubMed

Perina, Fabiano J; Amaral, Douglas C; Fernandes, Rafael S; Labory, Claudia Rg; Teixeira, Glauco A; Alves, Eduardo

2015-10-01

In initial assays, Thymus vulgaris essential oil (TEO) has demonstrated activity against several plant-pathogenic fungi and has reduced the fungal diseases to levels comparable with commercial fungicides. Thus, the goal of this work was to identify the mode of action in fungi of TEO and its major compound thymol (TOH) at the cellular level using an ultrastructure approach. TEO from leaves and TOH had minimum inhibitory concentrations (MICs) of 500 and 250 µg mL(-1) respectively against A. alternata; under the same conditions, MIC for a commercial fungicide was 1250 µg mL(-1) . Ultrastructure analysis showed that TOH phenolic substance prevented fungal growth, reduced fungal viability and prevented the penetration in fruits by a cell wall/plasma membrane interference mode of action with organelles targeted for destruction in the cytoplasm. Such mode of action differs from protective and preventive-curative commercial fungicides used as pattern control. These findings suggest that TOH was responsible for the antifungal activity of TEO. Therefore, both the essential oil and its major substance have potential for use in the development of new phenolic structures and analogues to control Alternaria brown spot disease caused by Alternaria alternata. © 2014 Society of Chemical Industry.

Contribution of a harpin protein from Xanthomonas axonopodis pv. citri to pathogen virulence.

PubMed

Sgro, Germán G; Ficarra, Florencia A; Dunger, Germán; Scarpeci, Telma E; Valle, Estela M; Cortadi, Adriana; Orellano, Elena G; Gottig, Natalia; Ottado, Jorgelina

2012-12-01

Xanthomonas axonopodis pv. citri (Xac), the bacterium that causes citrus canker, contains a gene in the hrp [for hypersensitive response (HR) and pathogenicity] cluster that encodes a harpin protein called Hpa1. Hpa1 produced HR in the nonhost plants tobacco, pepper and Arabidopsis, whereas, in the host plant citrus, it elicited a weak defence response with no visible phenotype. Co-infiltrations of Xac with or without the recombinant Hpa1 protein in citrus leaves produced a larger number of cankers in the presence of the protein. To characterize the effect of Hpa1 during the disease, an XacΔhpa1 mutant was constructed, and infiltration of this mutant caused a smaller number of cankers. In addition, the lack of Hpa1 hindered bacterial aggregation both in solution and in planta. Analysis of citrus leaves infiltrated with Hpa1 revealed alterations in mesophyll morphology caused by the presence of cavitations and crystal idioblasts, suggesting the binding of the harpin to plant membranes and the elicitation of signalling cascades. Overall, these results suggest that, even though Hpa1 elicits the defence response in nonhost plants and, to a lesser extent, in host plants, its main roles in citrus canker are to alter leaf mesophyll structure and to aggregate bacterial cells, and thus increase virulence and pathogen fitness. We expressed the N-terminal and C-terminal regions and found that, although both regions elicited HR in nonhost plants, only the N-terminal region showed increased virulence and bacterial aggregation, supporting the role of this region of the protein as the main active domain. © 2012 THE AUTHORS. MOLECULAR PLANT PATHOLOGY © 2012 BSPP AND BLACKWELL PUBLISHING LTD.

Evaluation of Alternaria mycotoxins in strawberries: quantification and storage condition.

PubMed

Juan, Cristina; Oueslati, Souheib; Mañes, Jordi

2016-05-01

Alternariol (AOH), alternariol methyl ether (AME) and tentoxin (TEN) are Alternaria mycotoxins produced by the most common post-harvest pathogens of fruits. The production of these metabolites depends on several environmental factors, mainly temperature, water activity, pH and the technological treatments that have been applied to the product. In this study, the occurrence of AOH, AME and TEN was evaluated in strawberries samples stored at different temperatures ranges (at 22 ± 2 or 6 ± 2°C) and different periods (up to 1 month) simulating the current practice of consumer's storage conditions. Sample extraction was performed using a liquid-liquid extraction method prior to LC-MS/MS analysis. AOH was the most prevalent mycotoxins with a 42% at strawberries stored at (22 ± 2)°C and 37% stored at (6 ± 2)°C. The highest AOH levels were found in samples conserved at (22 ± 2)°C ranging between 26 and 752 ng g(-1). AME levels ranged between 11 and 137 ng g(-)(1), which were found mainly in stored samples at (6 ± 2)°C for more than 28 days. None sample presented levels of TEN in either of the studied conditions.

[Climatic suitability of citrus in subtropical China].

PubMed

Duan, Hai-Lai; Qian, Huai-Sui; Li, Ming-Xia; Du, Yao-Dong

2010-08-01

By applying the theories of ecological suitability and the methods of fuzzy mathematics, this paper established a climatic suitability model for citrus, calculated and evaluated the climatic suitability and its spatiotemporal differences for citrus production in subtropical China, and analyzed the climatic suitability of citrus at its different growth stages and the mean climatic suitability of citrus in different regions of subtropical China. The results showed that the citrus in subtropical China had a lower climatic suitability and a higher risk at its flower bud differentiation stage, budding stage, and fruit maturity stage, but a higher climatic suitability and a lower risk at other growth stages. Cold damage and summer drought were the key issues affecting the citrus production in subtropical China. The citrus temperature suitability represented a latitudinal zonal pattern, i. e., decreased with increasing latitude; its precipitation suitability was high in the line of "Sheyang-Napo", medium in the southeast of the line, low in the northwest of the line, and non in high mountainous area; while the sunlight suitability was in line with the actual duration of sunshine, namely, higher in high-latitude areas than in low-latitude areas, and higher in high-altitude areas than in plain areas. Limited by temperature factor, the climatic suitability was in accordance with temperature suitability, i. e., south parts had a higher suitability than north parts, basically representing latitudinal zonal pattern. From the analysis of the inter-annual changes of citrus climatic suitability, it could be seen that the citrus climatic suitability in subtropical China was decreasing, and had obvious regional differences, suggesting that climate change could bring about the changes in the regions suitable for citrus production and in the key stages of citrus growth.

Sensitization to Alternaria and Cladosporium in patients with respiratory allergy and outdoor counts of mold spores in Ankara atmosphere, Turkey.

PubMed

Bavbek, Sevim; Erkekol, Ferda Oner; Ceter, Talip; Mungan, Dilşad; Ozer, Faruk; Pinar, Münevver; Misirligil, Zeynep

2006-08-01

Sensitization to Alternaria and Cladosporium has been reported to be 3% to 30% in European countries. However, in Turkey, there is limited data about the prevalence of sensitization to these molds and the intensity of the two mold spores in Ankara atmosphere. This study was designed to evaluate the sensitization to Alternaria and Cladosporium in patients with respiratory allergy in Ankara and also the concentration of the two molds in Ankara atmosphere. Allergic rhinitis and asthma patients living in Ankara were included in the study. Demographic and diagnostic data of the patients were recorded. A skin prick test with extracts supplied by three different laboratories was used to evaluate the sensitization to Alternaria and Cladosporium. Mold spores were measured using a Burkard 7-day recording volumetric spore trap in Ankara atmosphere during a year. Overall sensitization to the two molds was found to be 14.8%, and isolated Alternaria or Cladosporiumsensitization was 3%. Considering the positive reaction to at least one of the three suppliers, the sensitization rate was 11.9% and 8.1% for Alternaria and Cladosporium, respectively. Cochran's Q homogenization test demonstrated that the positive and negative reaction were not homogeneous among three laboratories. The total number of mold spores in Ankara atmosphere was 429,264 spores/m3 of which 75.5% and 6% were constituted by Cladosporium and Alternaria, respectively. The prevalence of Cladosporium and Alternaria sensitization in respiratory allergy patients is quite similar to European countries; however, our data indicate that commercial mold extracts should be standardized to establish the real sensitization rates. Additionally, considering the great numbers of these mold spores in Ankara atmosphere, long-term follow-up studies are needed to evaluate the relationship between the mold load and sensitization patterns.

Viability of and Escherichia coli O157:H7 and Listeria monocytogenes in a delicatessen appetizer (yogurt-based) salad as affected by citrus extract (Citrox©) and storage temperature.

PubMed

Tsiraki, Maria I; Yehia, Hany M; Elobeid, Tahra; Osaili, Tareq; Sakkas, Hercules; Savvaidis, Ioannis N

2018-02-01

The antimicrobial effect of citrus extract (at 1 mL/kg [C1] and 2 mL/kg [C2]) on naturally occurring microbiota and inoculated pathogens (E. coli O157:H7 and L. monocytogenes at ca. 6 log cfu/g) in the traditional Greek yogurt-based salad Tzatziki stored at 4, 10, or 21 °C, was examined. Lactic acid bacteria (LAB) were high (8.0-8.5 log cfu/g) and varied only minimally for both the control (untreated) and the citrus extract-treated salad samples, whereas the higher citrus extract concentration yielded the lowest yeast populations, irrespective of temperature, during the entire storage period. Populations of inoculated E. coli (6 log cfu/g) declined in both untreated and citrus extract-treated samples from day 0-70, 35, and 15 at 4, 10, and 21 °C, respectively. Citrus extract had a significant effect on the survival of the inoculated E. coli O157:H7, with reductions of 2.8-4.8 log cfu/g in the citrus extract-treated samples at the end of the storage period. Our data show that L. monocytogenes survived in both untreated and citrus extract-treated samples during the entire storage period, irrespective of the storage temperature. The higher concentration of citrus extract had a significant effect on the survival of L. monocytogenes in the treated samples, and reductions of 1.5-3.0 logs were noted on final day 70, 35 and 15 at 4, 10 and 21 °C, respectively. The results of our study demonstrated the potential of citrus extract as a natural compound that can control the growth of food-borne pathogenic bacteria, such as E. coli O157:H7 and L. monocytogenes in Tzatziki, a yogurt-based salad. Copyright © 2017 Elsevier Ltd. All rights reserved.

Yield and quality responses of citrus (Citrus reticulate) and tea (Podocarpus fleuryi Hickel.) to compound fertilizers*

PubMed Central

Wang, Rui; Shi, Xue-gen; Wei, You-zhang; Yang, Xiao-e; Uoti, Juhani

2006-01-01

Experiments were carried out with citrus (Citrus reticulate) and tea (Podocarpus fleuryi Hickel.) to study the effects of compound fertilizers on their yields and quality. In the citrus experiment, application of compound fertilizers increased available P, K and Mg contents in soil but decreased alkali-hydrolyzable N contents in soil and N, P and K contents in leaves. In the tea experiment, application of compound fertilizers increased available P, K and Mg contents in soil and N, P, K and Mg contents in leaves but decreased alkali-hydrolyzable N in soil compared with the urea treatment. Application of compound fertilizers could improve the quality of citrus and tea, increase their yields and enhance their economical profits significantly. Compared with the control, application of compound fertilizers increased citrus yields by 6.31, 12.94 and 17.69 t/ha, and those of tea by 0.51, 0.86 and 1.30 t/ha, respectively. Correspondingly, profits were increased by 21.4% to 61.1% for citrus and by 10.0% to 15.7% for tea. Optimal rates of compound fertilizers were recommended for both crops. PMID:16909469

Spectral sensitivity of the Asian citrus psyllid, Diaphorina citri

USDA-ARS?s Scientific Manuscript database

The Asian Citrus psyllid, Diaphorina citri, as a vector of the bacteria causing citrus greening, is considered one of the most important citrus pests globally. Movement of infected psyllids onto uninfected young citrus remains a key concern for the maintenance of citrus production. Attraction of d...

The Pathogenicity Determinant of Citrus Tristeza Virus Causing the Seedling Yellows Syndrome is Located at the 3’-Terminal Region of the Viral Genome

USDA-ARS?s Scientific Manuscript database

Citrus tristeza virus (CTV) (genus Closterovirus, family Closteroviridae) causes some of the more important viral diseases of citrus worldwide. The ability to map disease-inducing determinants of CTV is needed to develop better diagnostic and disease control procedures. A distinctive phenotype of s...

Novel dimeric metabolites from Alternaria tagetica.

PubMed

Gamboa-Angulo, M M; Alejos-González, F; Escalante-Erosa, F; García-Sosa, K; Delgado-Lamas, G; Peña-Rodríguez, L M

2000-08-01

Two novel polyketides, bis-7-O-8' '.8-O-7' '- and bis-7-O-7' '. 8-O-8' '-zinniol (2 and 3, respectively) were isolated from the organic crude extract of culture filtrates from Alternaria tagetica. Both structures were determined on the basis of their spectroscopic data (IR, MS, (1)H NMR, (13)C NMR, and 2D NMR experiments) and confirmed by chemical synthesis. Zinniol (1) was isolated as a major component, and its (13)C NMR data was correctly assigned after careful analysis of data from its 2D NMR experiments (HMQC and HMBC).

Invasive mutualisms between a plant pathogen and insect vectors in the Middle East and Brazil

PubMed Central

Queiroz, Renan Batista; Silva, Fábio Nascimento; Al-Mahmmoli, Issa Hashil; Al-Sadi, Abdullah Mohammed; Carvalho, Claudine Márcia; Elliot, Simon L.

2016-01-01

Complex multi-trophic interactions in vectorborne diseases limit our understanding and ability to predict outbreaks. Arthropod-vectored pathogens are especially problematic, with the potential for novel interspecific interactions during invasions. Variations and novelties in plant–arthropod–pathogen triumvirates present significant threats to global food security. We examined aspects of a phytoplasma pathogen of citrus across two continents. ‘Candidatus Phytoplasma aurantifolia’ causes Witches' Broom Disease of Lime (WBDL) and has devastated citrus production in the Middle East. A variant of this phytoplasma currently displays asymptomatic or ‘silent’ infections in Brazil. We first studied vector capacity and fitness impacts of the pathogen on its vectors. The potential for co-occurring weed species to act as pathogen reservoirs was analysed and key transmission periods in the year were also studied. We demonstrate that two invasive hemipteran insects—Diaphorina citri and Hishimonus phycitis—can vector the phytoplasma. Feeding on phytoplasma-infected hosts greatly increased reproduction of its invasive vector D. citri both in Oman and Brazil; suggesting that increased fitness of invasive insect vectors thereby further increases the pathogen's capacity to spread. Based on our findings, this is a robust system for studying the effects of invasions on vectorborne diseases and highlights concerns about its spread to warmer, drier regions of Brazil. PMID:28083099

Citrus breeding, genetics and genomics in Japan

PubMed Central

Omura, Mitsuo; Shimada, Takehiko

2016-01-01

Citrus is one of the most cultivated fruits in the world, and satsuma mandarin (Citrus unshiu Marc.) is a major cultivated citrus in Japan. Many excellent cultivars derived from satsuma mandarin have been released through the improvement of mandarins using a conventional breeding method. The citrus breeding program is a lengthy process owing to the long juvenility, and it is predicted that marker-assisted selection (MAS) will overcome the obstacle and improve the efficiency of conventional breeding methods. To promote citrus molecular breeding in Japan, a genetic mapping was initiated in 1987, and the experimental tools and resources necessary for citrus functional genomics have been developed in relation to the physiological analysis of satsuma mandarin. In this paper, we review the progress of citrus breeding and genome researches in Japan and report the studies on genetic mapping, expression sequence tag cataloguing, and molecular characterization of breeding characteristics, mainly in terms of the metabolism of bio-functional substances as well as factors relating to, for example, fruit quality, disease resistance, polyembryony, and flowering. PMID:27069387

Innate immune system capabilities of the Asian citrus psyllid, Diaphorina citri.

PubMed

Arp, Alex P; Martini, Xavier; Pelz-Stelinski, Kirsten S

2017-09-01

Citrus production worldwide is currently threatened by Huanglongbing, or citrus greening disease. The associated pathogen, Candidatus Liberibacter asiaticus (CLas), is transmitted by the Asian citrus psyllid, Diaphorina citri. Annotation of the D. citri genome revealed a reduced innate immune system lacking a number of antimicrobial peptides and the Imd pathway associated with defense against Gram-negative bacteria. We characterized this apparent immune reduction in survival assays in which D. citri were exposed to Gram-negative or Gram-positive bacteria. D. citri experienced significant mortality when exposed to Serratia marcescens (Gram-negative) through oral ingestion or by septic injury. Escherichia coli (Gram-negative) also caused significant D. citri mortality, but only when inoculated at high concentrations through oral ingestion or by septic injury. Neither Micrococcus luteus (Gram-positive) or Bacillus subtilis (Gram-positive) caused significant mortality as compared to controls in any experiment. E. coli titers increased rapidly following exposure, while M. luteus titer remained stable for 72 h. We demonstrate that D. citri is capable of defending against E. coli, a Gram-negative bacterium, despite lacking the Imd defense pathway. The tolerance of D. citri to M. luteus infection, yet inability to effectively clear infections, presents questions to efficacy of D. citri immune response to effectively clear Gram-positive infections. Copyright © 2017. Published by Elsevier Inc.

Antimicrobial Nanoemulsion Formulation with Improved Penetration of Foliar Spray through Citrus Leaf Cuticles to Control Citrus Huanglongbing.

PubMed

Yang, Chuanyu; Powell, Charles A; Duan, Yongping; Shatters, Robert; Zhang, Muqing

2015-01-01

Huanglongbing (HLB) is the most serious disease affecting the citrus industry worldwide to date. The causal agent, Candidatus Liberibacter asiaticus (Las), resides in citrus phloem, which makes it difficult to effectively treat with chemical compounds. In this study, a transcuticular nanoemulsion formulation was developed to enhance the permeation of an effective antimicrobial compound (ampicillin; Amp) against HLB disease through the citrus cuticle into the phloem via a foliar spray. The results demonstrated that efficiency of cuticle isolation using an enzymatic method (pectinase and cellulase) was dependent on the citrus cultivar and Las-infection, and it was more difficult to isolate cuticles from valencia orange (Citrus sinensis) and HLB-symptomatic leaves. Of eight adjuvants tested, Brij 35 provided the greatest increase in permeability of the HLB-affected cuticle with a 3.33-fold enhancement of cuticular permeability over water control. An in vitro assay using Bacillus subtilis showed that nanoemulsion formulations containing Amp (droplets size = 5.26 ± 0.04 nm and 94 ± 1.48 nm) coupled with Brij 35 resulted in greater inhibitory zone diameters (5.75 mm and 6.66 mm) compared to those of Brij 35 (4.34 mm) and Amp solution (2.83 mm) alone. Furthermore, the nanoemulsion formulations eliminated Las bacteria in HLB-affected citrus in planta more efficiently than controls. Our study shows that a water in oil (W/O) nanoemulsion formulation may provide a useful model for the effective delivery of chemical compounds into citrus phloem via a foliar spray for controlling citrus HLB.

Antimicrobial Nanoemulsion Formulation with Improved Penetration of Foliar Spray through Citrus Leaf Cuticles to Control Citrus Huanglongbing

PubMed Central

Yang, Chuanyu; Powell, Charles A.; Duan, Yongping; Shatters, Robert; Zhang, Muqing

2015-01-01

Huanglongbing (HLB) is the most serious disease affecting the citrus industry worldwide to date. The causal agent, Candidatus Liberibacter asiaticus (Las), resides in citrus phloem, which makes it difficult to effectively treat with chemical compounds. In this study, a transcuticular nanoemulsion formulation was developed to enhance the permeation of an effective antimicrobial compound (ampicillin; Amp) against HLB disease through the citrus cuticle into the phloem via a foliar spray. The results demonstrated that efficiency of cuticle isolation using an enzymatic method (pectinase and cellulase) was dependent on the citrus cultivar and Las-infection, and it was more difficult to isolate cuticles from valencia orange (Citrus sinensis) and HLB-symptomatic leaves. Of eight adjuvants tested, Brij 35 provided the greatest increase in permeability of the HLB-affected cuticle with a 3.33-fold enhancement of cuticular permeability over water control. An in vitro assay using Bacillus subtilis showed that nanoemulsion formulations containing Amp (droplets size = 5.26 ± 0.04 nm and 94 ± 1.48 nm) coupled with Brij 35 resulted in greater inhibitory zone diameters (5.75 mm and 6.66 mm) compared to those of Brij 35 (4.34 mm) and Amp solution (2.83 mm) alone. Furthermore, the nanoemulsion formulations eliminated Las bacteria in HLB-affected citrus in planta more efficiently than controls. Our study shows that a water in oil (W/O) nanoemulsion formulation may provide a useful model for the effective delivery of chemical compounds into citrus phloem via a foliar spray for controlling citrus HLB. PMID:26207823

Changes in Anthocyanin Production during Domestication of Citrus.

PubMed

Butelli, Eugenio; Garcia-Lor, Andrés; Licciardello, Concetta; Las Casas, Giuseppina; Hill, Lionel; Recupero, Giuseppe Reforgiato; Keremane, Manjunath L; Ramadugu, Chandrika; Krueger, Robert; Xu, Qiang; Deng, Xiuxin; Fanciullino, Anne-Laure; Froelicher, Yann; Navarro, Luis; Martin, Cathie

2017-04-01

Mandarin ( Citrus reticulata ), citron ( Citrus medica ), and pummelo ( Citrus maxima ) are important species of the genus Citrus and parents of the interspecific hybrids that constitute the most familiar commercial varieties of Citrus : sweet orange, sour orange, clementine, lemon, lime, and grapefruit. Citron produces anthocyanins in its young leaves and flowers, as do species in genera closely related to Citrus , but mandarins do not, and pummelo varieties that produce anthocyanins have not been reported. We investigated the activity of the Ruby gene, which encodes a MYB transcription factor controlling anthocyanin biosynthesis, in different accessions of a range of Citrus species and in domesticated cultivars. A white mutant of lemon lacks functional alleles of Ruby , demonstrating that Ruby plays an essential role in anthocyanin production in Citrus Almost all the natural variation in pigmentation by anthocyanins in Citrus species can be explained by differences in activity of the Ruby gene, caused by point mutations and deletions and insertions of transposable elements. Comparison of the allelic constitution of Ruby in different species and cultivars also helps to clarify many of the taxonomic relationships in different species of Citrus , confirms the derivation of commercial varieties during domestication, elucidates the relationships within the subgenus Papeda , and allows a new genetic classification of mandarins. © 2017 American Society of Plant Biologists. All Rights Reserved.

Enumerative and binomial sampling plans for citrus mealybug (Homoptera: pseudococcidae) in citrus groves.

PubMed

Martínez-Ferrer, María Teresa; Ripollés, José Luís; Garcia-Marí, Ferran

2006-06-01

The spatial distribution of the citrus mealybug, Planococcus citri (Risso) (Homoptera: Pseudococcidae), was studied in citrus groves in northeastern Spain. Constant precision sampling plans were designed for all developmental stages of citrus mealybug under the fruit calyx, for late stages on fruit, and for females on trunks and main branches; more than 66, 286, and 101 data sets, respectively, were collected from nine commercial fields during 1992-1998. Dispersion parameters were determined using Taylor's power law, giving aggregated spatial patterns for citrus mealybug populations in three locations of the tree sampled. A significant relationship between the number of insects per organ and the percentage of occupied organs was established using either Wilson and Room's binomial model or Kono and Sugino's empirical formula. Constant precision (E = 0.25) sampling plans (i.e., enumerative plans) for estimating mean densities were developed using Green's equation and the two binomial models. For making management decisions, enumerative counts may be less labor-intensive than binomial sampling. Therefore, we recommend enumerative sampling plans for the use in an integrated pest management program in citrus. Required sample sizes for the range of population densities near current management thresholds, in the three plant locations calyx, fruit, and trunk were 50, 110-330, and 30, respectively. Binomial sampling, especially the empirical model, required a higher sample size to achieve equivalent levels of precision.

Small RNA Profiling Reveals Phosphorus Deficiency as a Contributing Factor in Symptom Expression for Citrus Huanglongbing Disease

PubMed Central

Zhao, Hongwei; Sun, Ruobai; Jin, Hailing

2013-01-01

Huanglongbing (HLB) is a devastating citrus disease that is associated with bacteria of the genus ‘Candidatus Liberibacter’ (Ca. L.). Powerful diagnostic tools and management strategies are desired to control HLB. Host small RNAs (sRNA) play a vital role in regulating host responses to pathogen infection and are used as early diagnostic markers for many human diseases, including cancers. To determine whether citrus sRNAs regulate host responses to HLB, sRNAs were profiled from Citrus sinensis 10 and 14 weeks post grafting with Ca. L. asiaticus (Las)-positive or healthy tissue. Ten new microRNAs (miRNAs), 76 conserved miRNAs, and many small interfering RNAs (siRNAs) were discovered. Several miRNAs and siRNAs were highly induced by Las infection, and can be potentially developed into early diagnosis markers of HLB. miR399, which is induced by phosphorus starvation in other plant species, was induced specifically by infection of Las but not Spiroplasma citri that causes citrus stubborn—a disease with symptoms similar to HLB. We found a 35% reduction of phosphorus in Las-positive citrus trees compared to healthy trees. Applying phosphorus oxyanion solutions to HLB-positive sweet orange trees reduced HLB symptom severity and significantly improved fruit production during a 3-year field trial in south-west Florida. Our molecular, physiological, and field data suggest that phosphorus deficiency is linked to HLB disease symptomology. PMID:23292880

[Determination of Alternaria mycotoxins in selected raw and processed fruit and vegetable products].

PubMed

Giryn, H; Szteke, B

1995-01-01

The purpose of the study was the assessment of Alternaria-mycotoxins contamination in some raw and processed plant products. The analytical method for detection of alternariol (AOH) and alternariol methyl ether (AME) is described. After extraction and purification of sample crude extracts by column chromatography on silica gel, the qualitative and quantitative analyses were carried out by two-dimensional TLC. There were 110 samples analyzed -44 (included 32 moulded) raw plant samples and 56 processed plant products. Levels of alternaria-mycotoxins found in fruits and tomatoes visibly decayed ranged between 3 to 420 micrograms/kg for AOH and 10 to 100 micrograms/kg for AME. Trace amounts of AOH were detected in 3 samples of processed products.

Terpene Down-Regulation in Orange Reveals the Role of Fruit Aromas in Mediating Interactions with Insect Herbivores and Pathogens1[C][W

PubMed Central

Rodríguez, Ana; San Andrés, Victoria; Cervera, Magdalena; Redondo, Ana; Alquézar, Berta; Shimada, Takehiko; Gadea, José; Rodrigo, María Jesús; Zacarías, Lorenzo; Palou, Lluís; López, María M.; Castañera, Pedro; Peña, Leandro

2011-01-01

Plants use volatile terpene compounds as odor cues for communicating with the environment. Fleshy fruits are particularly rich in volatiles that deter herbivores and attract seed dispersal agents. We have investigated how terpenes in citrus fruit peels affect the interaction between the plant, insects, and microorganisms. Because limonene represents up to 97% of the total volatiles in orange (Citrus sinensis) fruit peel, we chose to down-regulate the expression of a limonene synthase gene in orange plants by introducing an antisense construct of this gene. Transgenic fruits showed reduced accumulation of limonene in the peel. When these fruits were challenged with either the fungus Penicillium digitatum or with the bacterium Xanthomonas citri subsp. citri, they showed marked resistance against these pathogens that were unable to infect the peel tissues. Moreover, males of the citrus pest medfly (Ceratitis capitata) were less attracted to low limonene-expressing fruits than to control fruits. These results indicate that limonene accumulation in the peel of citrus fruit appears to be involved in the successful trophic interaction between fruits, insects, and microorganisms. Terpene down-regulation might be a strategy to generate broad-spectrum resistance against pests and pathogens in fleshy fruits from economically important crops. In addition, terpene engineering may be important for studying the basic ecological interactions between fruits, herbivores, and pathogens. PMID:21525333

PdSlt2 Penicillium digitatum mitogen-activated-protein kinase controls sporulation and virulence during citrus fruit infection.

PubMed

de Ramón-Carbonell, Marta; Sánchez-Torres, Paloma

2017-12-01

The Slt2 mitogen-activated protein (MAP) kinase homologue of Penicillium digitatum, the most relevant pathogen-producing citrus green mould decay during postharvest, was identified and explored. The P. digitatum Slt2-MAPK coding gene (PdSlt2) was functionally characterized by homologous gene elimination and transcriptomic evaluation. The absence of PdSlt2 gene resulted in significantly reduced virulence during citrus infection. The ΔPdSlt2 mutants were also defective in asexual reproduction, showing impairment of sporulation during citrus infection. Gene expression analysis revealed that PdSlt2 was highly induced during citrus fruit infection at early stages (1 dpi). Moreover, PdSlt2 deletion altered gene expression profiles. The relative gene expression (RGE) of fungicide resistance- and fungal virulence-related genes showed that PdSlt2 acts as negative regulator of several transporter encoding genes (ABC and MFS transporters) and a positive regulator of two sterol demethylases. This study indicates that PdSlt2 MAPK is functionally preserved in P. digitatum and highlights the relevant role of the PdSlt2 MAP kinase-mediated signalling pathway in regulating diverse genes crucial for infection and asexual reproduction. Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Improved annotation of the insect vector of citrus greening disease: biocuration by a diverse genomics community

PubMed Central

Hosmani, Prashant S.; Villalobos-Ayala, Krystal; Miller, Sherry; Shippy, Teresa; Flores, Mirella; Rosendale, Andrew; Cordola, Chris; Bell, Tracey; Mann, Hannah; DeAvila, Gabe; DeAvila, Daniel; Moore, Zachary; Buller, Kyle; Ciolkevich, Kathryn; Nandyal, Samantha; Mahoney, Robert; Van Voorhis, Joshua; Dunlevy, Megan; Farrow, David; Hunter, David; Morgan, Taylar; Shore, Kayla; Guzman, Victoria; Izsak, Allison; Dixon, Danielle E.; Cridge, Andrew; Cano, Liliana; Cao, Xiaolong; Jiang, Haobo; Leng, Nan; Johnson, Shannon; Cantarel, Brandi L.; Richards, Stephen; English, Adam; Shatters, Robert G.; Childers, Chris; Chen, Mei-Ju; Hunter, Wayne; Cilia, Michelle; Mueller, Lukas A.; Munoz-Torres, Monica; Nelson, David; Poelchau, Monica F.; Benoit, Joshua B.; Wiersma-Koch, Helen; D’Elia, Tom; Brown, Susan J.

2017-01-01

Abstract The Asian citrus psyllid (Diaphorina citri Kuwayama) is the insect vector of the bacterium Candidatus Liberibacter asiaticus (CLas), the pathogen associated with citrus Huanglongbing (HLB, citrus greening). HLB threatens citrus production worldwide. Suppression or reduction of the insect vector using chemical insecticides has been the primary method to inhibit the spread of citrus greening disease. Accurate structural and functional annotation of the Asian citrus psyllid genome, as well as a clear understanding of the interactions between the insect and CLas, are required for development of new molecular-based HLB control methods. A draft assembly of the D. citri genome has been generated and annotated with automated pipelines. However, knowledge transfer from well-curated reference genomes such as that of Drosophila melanogaster to newly sequenced ones is challenging due to the complexity and diversity of insect genomes. To identify and improve gene models as potential targets for pest control, we manually curated several gene families with a focus on genes that have key functional roles in D. citri biology and CLas interactions. This community effort produced 530 manually curated gene models across developmental, physiological, RNAi regulatory and immunity-related pathways. As previously shown in the pea aphid, RNAi machinery genes putatively involved in the microRNA pathway have been specifically duplicated. A comprehensive transcriptome enabled us to identify a number of gene families that are either missing or misassembled in the draft genome. In order to develop biocuration as a training experience, we included undergraduate and graduate students from multiple institutions, as well as experienced annotators from the insect genomics research community. The resulting gene set (OGS v1.0) combines both automatically predicted and manually curated gene models. Database URL: https://citrusgreening.org/ PMID:29220441

Citrus fruit extracts with carvacrol and thymol eliminated 7-log acid-adapted Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes: A potential of effective natural antibacterial agents.

PubMed

Chung, Doohyun; Cho, Tae Jin; Rhee, Min Suk

2018-05-01

Despite the widespread belief that citrus fruit extracts (CFEs) are microbiologically safe due to their acidity, limited bactericidal effect results in low applicability as antibacterial agent and outbreaks occurred by acid-adapted pathogens. Here, we examined the antibacterial effects of CFEs [lime (Citrus medica), lemon (Citrus limon), calamansi (Citrus microcarpa)] combined with essential oil components (EOCs; carvacrol and thymol) against non-acid-adapted/acid-adapted Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes under 22 °C for 5 min. CFEs (<20%) alone or small amounts of EOCs (2.0 mM; 0.032%) alone could not inactivate the target bacteria effectively. However, combined treatments exhibited marked synergy: CFE + EOCs eliminated all the bacteria (>6.9 log CFU/ml). Among the CFEs tested, the highest synergism was shown by calamansi, an exotic citrus fruit previously unrecognized as an antibacterial agent. Although acid-adaptation improved bacterial survival, calamansi (<20%) + EOCs (<0.032%) completely inactivated even the most resistant pathogen (E. coli O157:H7). Validation test also showed that all tested commercial juice products also eliminated acid-adapted pathogens when used with EOCs. Physicochemical analysis of tested CFEs (pH measurement and HPLC analysis of components) revealed that low pH and flavanone (hesperidin) did not contribute to the synergistic bactericidal effects. Rather, the high citric acid content is likely to contribute to the strong synergistic effect with EOCs by damaging susceptible bacterial membranes. Sensory scores for CFEs were not altered by addition of EOCs at concentrations up to 1.5 mM. This study provides new insight into the utility of CFEs with EOCs to improve not only the microbiological safety of food products containing CFEs but also their applicability as natural antibacterial complex. Copyright © 2018 Elsevier Ltd. All rights reserved.

The aconitate hydratase family from Citrus

PubMed Central

2010-01-01

Background Research on citrus fruit ripening has received considerable attention because of the importance of citrus fruits for the human diet. Organic acids are among the main determinants of taste and organoleptic quality of fruits and hence the control of fruit acidity loss has a strong economical relevance. In citrus, organic acids accumulate in the juice sac cells of developing fruits and are catabolized thereafter during ripening. Aconitase, that transforms citrate to isocitrate, is the first step of citric acid catabolism and a major component of the citrate utilization machinery. In this work, the citrus aconitase gene family was first characterized and a phylogenetic analysis was then carried out in order to understand the evolutionary history of this family in plants. Gene expression analyses of the citrus aconitase family were subsequently performed in several acidic and acidless genotypes to elucidate their involvement in acid homeostasis. Results Analysis of 460,000 citrus ESTs, followed by sequencing of complete cDNA clones, identified in citrus 3 transcription units coding for putatively active aconitate hydratase proteins, named as CcAco1, CcAco2 and CcAco3. A phylogenetic study carried on the Aco family in 14 plant species, shows the presence of 5 Aco subfamilies, and that the ancestor of monocot and dicot species shared at least one Aco gene. Real-time RT-PCR expression analyses of the three aconitase citrus genes were performed in pulp tissues along fruit development in acidic and acidless citrus varieties such as mandarins, oranges and lemons. While CcAco3 expression was always low, CcAco1 and CcAco2 genes were generally induced during the rapid phase of fruit growth along with the maximum in acidity and the beginning of the acid reduction. Two exceptions to this general pattern were found: 1) Clemenules mandarin failed inducing CcAco2 although acid levels were rapidly reduced; and 2) the acidless "Sucreña" orange showed unusually high levels

Survey of Alternaria toxin contamination in food from the German market, using a rapid HPLC-MS/MS approach.

PubMed

Hickert, Sebastian; Bergmann, Marian; Ersen, Seyma; Cramer, Benedikt; Humpf, Hans-Ulrich

2016-02-01

A HPLC-MS/MS-based method for the quantification of nine mycotoxins produced by fungi of the genus Alternaria in various food matrices was developed. The method relies on a single-step extraction, followed by dilution of the raw extract and direct analysis. In combination with an analysis time per sample of 12 min, the sample preparation is cost-effective and easy to handle. The method covers alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA), altenuene (ALT), iso-altenuene (isoALT), tentoxin (TEN), altertoxin-I (ATX-I), and the AAL toxins TA1 and TA2. Some Alternaria toxins which are either not commercially available or very expensive, namely AOH, AME, ALT, isoALT, and ATX-I, were isolated as reference compounds from fungal cultures. The method was extensively validated for tomato products, bakery products, sunflower seeds, fruit juices, and vegetable oils. AOH, AME, TeA, and TEN were found in quantifiable amounts and 92.1% of all analyzed samples (n = 96) showed low level contamination with one or more Alternaria toxins. Based on the obtained results, the average daily exposure to Alternaria toxins in Germany was calculated.

Antibacterial activity of different honeys against pathogenic bacteria.

PubMed

Voidarou, C; Alexopoulos, A; Plessas, S; Karapanou, A; Mantzourani, I; Stavropoulou, E; Fotou, K; Tzora, A; Skoufos, I; Bezirtzoglou, E

2011-12-01

To study the antimicrobial activity of honey, 60 samples of various botanical origin were evaluated for their antimicrobial activities against 16 clinical pathogens and their respective reference strains. The microbiological quality of honeys and the antibiotic susceptibility of the various isolates were also examined. The bioassay applied for determining the antimicrobial effect employs the well-agar diffusion method and the estimation of minimum active dilution which produces a 1mm diameter inhibition zone. All honey samples, despite their origin (coniferous, citrus, thyme or polyfloral), showed antibacterial activity against the pathogenic and their respective reference strains at variable levels. Coniferous and thyme honeys showed the highest activity with an average minimum dilution of 17.4 and 19.2% (w/v) followed by citrus and polyfloral honeys with 20.8 and 23.8% respectively. Clinical isolates of Staphylococcus aureus subsp. aureus, Escherichia coli, Salmonella enterica subsp. Enterica, Streptococcus pyogenes, Bacillus cereus and Bacillus subtilis were proven to be up to 60% more resistant than their equal reference strains thus emphasizing the variability in the antibacterial effect of honey and the need for further research. Copyright © 2011 Elsevier Ltd. All rights reserved.

Transcriptional analysis of sweet orange trees co-infected with 'Candidatus Liberibacter asiaticus' and mild or severe strains of Citrus tristeza virus.

PubMed

Fu, Shimin; Shao, Jonathan; Paul, Cristina; Zhou, Changyong; Hartung, John S

2017-10-31

Citrus worldwide is threatened by huanglongbing (HLB) and tristeza diseases caused by 'Candidatus Liberibacter asiaticus' (CaLas) and Citrus tristeza virus (CTV). Although the pathogens are members of the α-proteobacteria and Closteroviridae, respectively, both are restricted to phloem cells in infected citrus and are transmitted by insect vectors. The response of sweet orange to single infection by either of these two pathogens has been characterized previously by global gene expression analysis. But because of the ubiquity of these pathogens where the diseases occur, co-infection by both pathogens is very common and could lead to increased disease severity based on synergism. We therefore co-inoculated sweet orange trees with CaLas and either a mild or a severe strain of CTV, and measured changes of gene expression in host plants. In plants infected with CaLas-B232, the overall alteration in gene expression was much greater in plants co-inoculated with the severe strain of CTV, B6, than when co-infected with the mild strain of CTV, B2. Plants co-infected with CaLas-B232 and either strain of CTV died but trees co-infected with CTV-B2 survived much longer than those co-infected with CTV-B6. Many important pathways were perturbed by both CTV-B2/CaLas-B232 and/or CTV-B6/CaLas-B232, but always more severely by CTV-B6/CaLas-B232. Genes related to cell wall modification and metal transport responded differently to infection by the pathogens in combination than by the same pathogens singly. The expressions of genes encoding phloem proteins and sucrose loading proteins were also differentially altered in response to CTV-B2 or CTV-B6 in combination with CaLas-B232, leading to different phloem environments in plants co-infected by CaLas and mild or severe CTV. Many host genes were expressed differently in response to dual infection as compared to single infections with the same pathogens. Interactions of the pathogens within the host may lead to a better or worse result

The Efficacy and Underlying Mechanism of Sulfone Derivatives Containing 1,3,4-oxadiazole on Citrus Canker.

PubMed

Li, Pei; Ma, Yuhua; Zhou, Junliang; Luo, Hui; Yan, Jiawen; Mao, Yongya; Wang, Zhuang

2015-08-04

The objectives of the current study were to isolate and identify the pathogen responsible for citrus canker and investigate the efficacy of sulfone derivatives containing 1,3,4-oxadiazole moiety on controlling citrus canker caused by Xanthomonas citri subsp. citri (Xcc) under in vitro and field conditions. In an in vitro study, we tested eight sulfone derivatives against Xcc and the results demonstrated that compound 3 exhibited the best antibacterial activity against Xcc, with a half-maximal effective concentration (EC50) value of 1.23 μg/mL, which was even better than those of commercial bactericides Kocide 3000 (58.21 μg/mL) and Thiodiazole copper (77.04 μg/mL), respectively. Meanwhile, under field experiments, compound 3 treatments demonstrated the highest ability to reduce the disease of citrus canker in leaves and fruits in two different places relative to an untreated control as well as the commercial bactericides Kocide 3000 and Thiodiazole copper. Meanwhile, compound 3 could stimulate the increase in peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia lyase (PAL) activities in the navel orange leaves, causing marked enhancement of plant resistance against citrus canker. Moreover, compound 3 could damage the cell membranes, destruct the biofilm formation, inhibit the production of extracellular polysaccharide (EPS), and affect the cell membrane permeability to restrain the growth of the bacteria.

Genomic Analysis of Terpene Synthase Family and Functional Characterization of Seven Sesquiterpene Synthases from Citrus sinensis

PubMed Central

Alquézar, Berta; Rodríguez, Ana; de la Peña, Marcos; Peña, Leandro

2017-01-01

Citrus aroma and flavor, chief traits of fruit quality, are derived from their high content in essential oils of most plant tissues, including leaves, stems, flowers, and fruits. Accumulated in secretory cavities, most components of these oils are volatile terpenes. They contribute to defense against herbivores and pathogens, and perhaps also protect tissues against abiotic stress. In spite of their importance, our understanding of the physiological, biochemical, and genetic regulation of citrus terpene volatiles is still limited. The availability of the sweet orange (Citrus sinensis L. Osbeck) genome sequence allowed us to characterize for the first time the terpene synthase (TPS) family in a citrus type. CsTPS is one of the largest angiosperm TPS families characterized so far, formed by 95 loci from which just 55 encode for putative functional TPSs. All TPS angiosperm families, TPS-a, TPS-b, TPS-c, TPS-e/f, and TPS-g were represented in the sweet orange genome, with 28, 18, 2, 2, and 5 putative full length genes each. Additionally, sweet orange β-farnesene synthase, (Z)-β-cubebene/α-copaene synthase, two β-caryophyllene synthases, and three multiproduct enzymes yielding β-cadinene/α-copaene, β-elemene, and β-cadinene/ledene/allo-aromandendrene as major products were identified, and functionally characterized via in vivo recombinant Escherichia coli assays. PMID:28883829

Widespread applications of citrus cryopreservation

USDA-ARS?s Scientific Manuscript database

Citrus genetic resources can now be successfully cryopreserved, which means that they can be placed into long-term storage at liquid nitrogen temperatures. This cryopreservation technology was specifically developed to address the immediate need to have secure long-term back-up storage for citrus co...

The flavor of citrus fruit

USDA-ARS?s Scientific Manuscript database

Citrus is the largest cultivated fruit tree crop in the world, with total production of more than 100 million tons per year. The genus Citrus consists of different species, including several producing economically important crops, such as oranges, mandarins, grapefruit, pummelo, lemons and limes, c...

Genome wide selection in Citrus breeding.

PubMed

Gois, I B; Borém, A; Cristofani-Yaly, M; de Resende, M D V; Azevedo, C F; Bastianel, M; Novelli, V M; Machado, M A

2016-10-17

Genome wide selection (GWS) is essential for the genetic improvement of perennial species such as Citrus because of its ability to increase gain per unit time and to enable the efficient selection of characteristics with low heritability. This study assessed GWS efficiency in a population of Citrus and compared it with selection based on phenotypic data. A total of 180 individual trees from a cross between Pera sweet orange (Citrus sinensis Osbeck) and Murcott tangor (Citrus sinensis Osbeck x Citrus reticulata Blanco) were evaluated for 10 characteristics related to fruit quality. The hybrids were genotyped using 5287 DArT_seq TM (diversity arrays technology) molecular markers and their effects on phenotypes were predicted using the random regression - best linear unbiased predictor (rr-BLUP) method. The predictive ability, prediction bias, and accuracy of GWS were estimated to verify its effectiveness for phenotype prediction. The proportion of genetic variance explained by the markers was also computed. The heritability of the traits, as determined by markers, was 16-28%. The predictive ability of these markers ranged from 0.53 to 0.64, and the regression coefficients between predicted and observed phenotypes were close to unity. Over 35% of the genetic variance was accounted for by the markers. Accuracy estimates with GWS were lower than those obtained by phenotypic analysis; however, GWS was superior in terms of genetic gain per unit time. Thus, GWS may be useful for Citrus breeding as it can predict phenotypes early and accurately, and reduce the length of the selection cycle. This study demonstrates the feasibility of genomic selection in Citrus.

Inhibition of spore germination of Alternaria tenuis by sulfur dioxide

DOE Office of Scientific and Technical Information (OSTI.GOV)

Couey, H.M.

1962-08-01

As a part of a continuing study of SO/sub 2/ fumigation of table grapes, the effect of SO/sub 2/ on spores of an isolate of A. tenuis Auct. causing decay of table grapes was determined. The amount of SO/sub 2/ required to inhibit completely spore germination depended on availability of moisture and the temperature. At 20/sup 0/C, wet spores required 20-min exposure to 100 ppm SO/sub 2/ to prevent germination, but spores equilibrated at 90% relative humidity (RH) required 10-min exposure to 1000 ppm SO/sub 2/. Dry spores at 60% RH were unaffected by a 20-min exposure to 4000 ppmmore » SO/sub 2/. Increasing the temperature in the range 5-20/sup 0/C increased effectiveness of the SO/sub 2/ treatment. A comparison of Alternaria with Botrytis cinerea Fr. (studied earlier) showed that wet spores of these organisms were about equally sensitive to SO/sub 2/, but that dry Alternaria spores were more resistant to SO/sub 2/ than dry Botrytis spores under comparable conditions.« less

Phenology of Asian citrus psyllid (Hemiptera: Liviidae) and associated parasitoids on two species of Citrus, kinnow mandarin and sweet orange, in Punjab Pakistan.

PubMed

Khan, Shouket Zaman; Arif, Muhammad Jalal; Hoddle, Christina D; Hoddle, Mark S

2014-10-01

The population phenology of Asian citrus psyllid, Diaphorina citri Kuwayama, was monitored weekly for 110 wk on two species of Citrus, kinnow mandarin and sweet orange, at two different research sites in Faisalabad, Punjab Pakistan. Citrus flush growth patterns were monitored and natural enemy surveys were conducted weekly. Flush patterns were similar for kinnow and sweet orange. However, flush on sweet orange was consistently more heavily infested with Asian citrus psyllid than kinnow flush; densities of Asian citrus psyllid eggs, nymphs, and adults were higher on sweet orange when compared with kinnow. When measured in terms of mean cumulative insect or Asian citrus psyllid days, eggs, nymphs, and adults were significantly higher on sweet orange than kinnow. Two parasitoids were recorded attacking Asian citrus psyllid nymphs, Tamarixia radiata (Waterston) and Diaphorencyrtus aligarhensis (Shafee, Alam and Agarwal). The dominant parasitoid species attacking Asian citrus psyllid nymphs on kinnow and sweet orange was T. radiata, with parasitism averaging 26%. D. aligarhensis parasitism averaged 17%. Generalist predators such as coccinellids and chrysopids were collected infrequently and were likely not important natural enemies at these study sites. Immature spiders, in particular, salticids and yellow sac spiders, were common and may be important predators of all Asian citrus psyllid life stages. Low year round Asian citrus psyllid densities on kinnow and possibly high summer temperatures, may, in part, contribute to the success of this cultivar in Punjab where Candidatus Liberibacter asiaticus, the putative causative agent of huanglongbing, a debilitating citrus disease, is widespread and vectored by Asian citrus psyllid.

Induction of gentisic acid 5-O-beta-D-xylopyranoside in tomato and cucumber plants infected by different pathogens.

PubMed

Fayos, Joaquín; Bellés, José María; López-Gresa, M Pilar; Primo, Jaime; Conejero, Vicente

2006-01-01

Tomato plants infected with the citrus exocortis viroid exhibited strongly elevated levels of a compound identified as 2,5-dihydroxybenzoic acid (gentisic acid, GA) 5-O-beta-D-xylopyranoside. The compound accumulated early in leaves expressing mild symptoms from both citrus exocortis viroid-infected tomato, and prunus necrotic ringspot virus-infected cucumber plants, and progressively accumulated concomitant with symptom development. The work presented here demonstrates that GA, mainly associated with systemic infections in compatible plant-pathogen interactions [Bellés, J.M., Garro, R., Fayos, J., Navarro, P., Primo, J., Conejero, V., 1999. Gentisic acid as a pathogen-inducible signal, additional to salicylic acid for activation of plant defenses in tomato. Mol. Plant-Microbe Interact. 12, 227-235], is conjugated to xylose. Notably, this result contrasts with those previously found in other plant-pathogen interactions in which phenolics analogues of GA as benzoic or salicylic acids, are conjugated to glucose.

In Vitro Assessment of Extracts of the Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Higher Basidiomycetes) Against Different Plant Pathogenic Fungi.

PubMed

Baig, Mirza Nabeel; Shahid, Ahmad Ali; Ali, Muhammad

2015-01-01

Five isolates of the lingzhi or reishi medicinal mushroom Ganoderma lucidum (GL-1, GL-2, GL-3, GL-4, GL-5) were collected from different locations within and surrounding Lahore, Pakistan, to study the antifungal potential of their bioactive compounds. After studying morphology, different concentrations of the extracts were prepared in methanol and water using a Soxhlet extractor. Different cultures of fungal pathogens were acquired from the First Fungal Culture Bank of Pakistan, University of the Punjab, Lahore. The antimicrobial potential of 5 G. lucidum samples against 5 fungal pathogens (Fusarium oxysporum, Aspergillus niger, A. flavus, Penicillium sp., and Alternaria alternata) was observed. The lowest biomass reduction (7%) was observed in 1% and 2% concentrations of a methanolic extract and 6% in the case of a water extract. Major inhibition was observed using higher concentrations of the methanolic extract (3% and 4%). These extracts significantly suppressed fungal biomass up to 38% and 56% in A. niger, 47% in A. flavus, 58% in ,i>Penicillium sp., 46% in A. alternaria, and 45% in F. oxysporum compared with the control. It was concluded from these studies that methanolic extracts of G. lucidum showed better activity against all plant fungal pathogens when compared with the water extracts.

Integrated Management of Citrus Canker

USDA-ARS?s Scientific Manuscript database

Fruit losses due to citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), vary each crop season depending on citrus variety, tree age, flushing condition, leafminer control, and coincidence of weather events with occurrence of susceptible fruit and foliage. In 2012, crop losses in Hamlin f...

H NMR analyses of Citrus macrophylla subjected to Asian citrus psyllid (Diaphorina citri Kuwayama) feeding

USDA-ARS?s Scientific Manuscript database

The Asian citrus psyllid (ACP) is a phloem feeding insect that can host and transmit the bacterium Candidatus Liberibacter asiaticus (CLas), which is the putative causative agent of the economically important citrus disease, Huanglongbing (HLB). ACP are widespread in Florida, and are spreading in Ca...

Isolation and characterization of the grain mold fungi, Cochliobolus and Alternaria spp., from sorghum using semi-selective media and DNA sequence analyses

USDA-ARS?s Scientific Manuscript database

Mold diseases, caused by fungal complexes including Alternaria, Cochliobolus and Fusarium species, limit sorghum grain production. Media were tested by plating Fusarium thapsinum, Alternaria sp. and Curvularia lunata, individually and competitively. Dichloran chloramphenicol rose bengal (DRBC) and m...

Comparative study of airborne Alternaria conidia levels in two cities in Castilla-La Mancha (central Spain), and correlations with weather-related variables.

PubMed

Sabariego, Silvia; Bouso, Veronica; Pérez-Badia, Rosa

2012-01-01

Alternaria conidia are among the airborne biological particles known to trigger allergic respiratory diseases. The presented paper reports on a study of seasonal variations in airborne Alternaria conidia concentrations in 2 cities in the central Spanish region of Castilla-La Mancha, Albacete and Toledo. The influence of weather-related variables on airborne conidia levels and distribution was also analysed. Sampling was carried out from 2008-2010 using a Hirst sampler, following the methodology established by the Spanish Aerobiology Network. Annual airborne Alternaria conidia counts were higher in Toledo (annual mean 3,936 conidia) than in Albacete (annual mean 2,268 conidia). Conidia were detected in the air throughout the year, but levels peaked between May-September. Considerable year-on-year variations were recorded both in total annual counts and in seasonal distribution. A significant positive correlation was generally found between mean daily Alternaria counts and both temperature and hours of sunlight, while a significant negative correlation was recorded for relative humidity, daily and cumulative rainfall, and wind speed. Regression models indicated that between 31%-52% of the variation in airborne Alternaria conidia concentrations could be explained by weather-related variables.

Isolating Fungal Pathogens from a Dynamic Disease Outbreak in a Native Plant Population to Establish Plant-Pathogen Bioassays for the Ecological Model Plant Nicotiana attenuata

PubMed Central

Schuck, Stefan; Baldwin, Ian T.

2014-01-01

The wild tobacco species Nicotiana attenuata has been intensively used as a model plant to study its interaction with insect herbivores and pollinators in nature, however very little is known about its native pathogen community. We describe a fungal disease outbreak in a native N. attenuata population comprising 873 plants growing in an area of about 1500 m2. The population was divided into 14 subpopulations and disease symptom development in the subpopulations was monitored for 16 days, revealing a waxing and waning of visible disease symptoms with some diseased plants recovering fully. Native fungal N. attenuata pathogens were isolated from diseased plants, characterized genetically, chemotaxonomically and morphologically, revealing several isolates of the ascomycete genera Fusarium and Alternaria, that differed in the type and strength of the disease symptoms they caused in bioassays on either detached leaves or intact soil-grown plants. These isolates and the bioassays will empower the study of N. attenuata-pathogen interactions in a realistic ecological context. PMID:25036191

RNAi-based strategy for Asian citrus psyllid (Diaphorina citri) Control: A method to reduce the spread of citrus greening disease

USDA-ARS?s Scientific Manuscript database

Citrus greening disease is a serious bacterial disease of citrus worldwide and is vectored by the Asian citrus pysllid (Diaphorina Citri). The only effective control strategy includes vigorous control of the psyllid, primarily through heavy reliance on pesticides. As a more sustainable and environm...

Antileukemic alpha-pyrone derivatives from the endophytic fungus Alternaria phragmospora

USDA-ARS?s Scientific Manuscript database

Four new (1–4) and two known (5 and 6)a-pyrone derivatives have been isolated from Alternaria phragmospora, an endophytic fungus from Vinca rosea, leaves. The isolated compounds were chemically identi'ed to be 5-butyl-4-methoxy-6-methyl-2H-pyran-2-one (2) 5-butyl-6-(hydroxymethyl)-4-methoxy-2H-py...

Salicylic acid treatment reduces the rot of postharvest citrus fruit by inducing the accumulation of H2O2, primary metabolites and lipophilic polymethoxylated flavones.

PubMed

Zhu, Feng; Chen, Jiajing; Xiao, Xue; Zhang, Mingfei; Yun, Ze; Zeng, Yunliu; Xu, Juan; Cheng, Yunjiang; Deng, Xiuxin

2016-09-15

To comprehensively analyze the effects of salicylic acid (SA) on the storability of Satsuma mandarin (Citrus unshiu), fruits were treated with 2mM SA. The disease incidence of control/SA-treated fruit at 50d and 120d after treatment was 23.3%/10% and 67.3%/23.3%, respectively, suggesting that SA treatment can significantly reduce the rot rate of postharvest citrus fruit. Fruit quality assays revealed that the treatment can maintain fruit firmness without affecting the inner quality. Furthermore, the contents of H2O2 and some defense-related metabolites, such as ornithine and threonine, in citrus pericarp, were significantly increased by SA treatment. Moreover, it was lipophilic polymethoxylated flavones, rather than flavanone glycosides, that accumulated in SA-treated fruits and these can directly inhibit pathogen development. These results suggest that the effects of SA on postharvest citrus fruit may be attributed to the accumulation of H2O2 and defense-related metabolites. Copyright © 2016. Published by Elsevier Ltd.

Development of SSR markers from Citrus clementina (Rutaceae) BAC end sequences and interspecific transferability in Citrus.

PubMed

Ollitrault, Frédérique; Terol, Javier; Pina, Jose Antonio; Navarro, Luis; Talon, Manuel; Ollitrault, Patrick

2010-11-01

Microsatellite primers were developed from bacterial artificial chromosome (BAC) end sequences of Citrus clementina and their transferability and polymorphism tested in the genus Citrus for future anchorage of physical and genetic maps and comparative interspecific genetic mapping. • Using PAGE and DNA silver staining, 79 primer pairs were selected for their transferability and polymorphism among 526 microsatellites mined in BES. A preliminary diversity study in Citrus was conducted with 18 of them, in C. reticulata, C. maxima, C. medica, C. sinensis, C. aurantium, C. paradisi, C. lemon, C. aurantifolia, and some papedas (wild citrus), using a capillary electrophoresis fragment analyzer. Intra- and interspecific polymorphism was observed, and heterozygous markers were identified for the different genotypes to be used for genetic mapping. • These results indicate the utility of the developed primers for comparative mapping studies and the integration of physical and genetic maps.

Herbivory by the insect diaphorina citri induces greater change in citrus plant volatile profile than does infection by the bacterium, Candidatus Liberibacter asiaticus.

PubMed

Hijaz, Faraj; El-Shesheny, Ibrahim; Killiny, Nabil

2013-10-01

The volatile organic compound (VOC) profile in plant leaves often changes after biotic and abiotic stresses. Monitoring changes in VOCs in plant leaves could provide valuable information about multitrophic interactions. In the current study, we investigated the effect of Asian citrus psyllid (ACP) infestation, citrus greening pathogen (Candidatus Liberibacter asiaticus [CLas]) infection, and simultaneous attack by ACP and CLas on the VOC content of citrus leaves. Leaf volatiles were extracted using hexane and analyzed with gas chromatography-mass spectrometry (GC-MS). Although ACP is a phloem-sucking insect that causes minimal damage to plant tissues, the relative amount of 21 out of the 27 VOCs increased 2- to 10-fold in ACP-infested plants. The relative amount of d-limonene, β-phelandrene, citronellal, and undecanal were increased 4- to 20- fold in CLas-infected plants. A principle component analysis (PCA) and cluster analysis (CA) showed that VOC patterns of ACP-infested and CLas-infected plants were different from each other and were also different from the controls, while the VOC pattern of double-attacked plants was more like that of the controls than that of ACP-infested or CLas-infected plants. VOC amounts from leaves were compromised when plants were attacked by ACP and CLas. The results of this study showed that a simple direct extraction of citrus leaf volatiles could be successfully used to discriminate between healthy and CLas-infected plants. Information about the effects of insect and pathogen attack on the VOC content profile of plants might contribute to a better understanding of biotic stress.

[Survey of sharpshooters (Hemiptera: Cicadellidae) associated with Xylella fastidiosa transmission in citrus groves of the North Coast of Bahia State].

PubMed

De Miranda, Marcelo P; Lopes, João R S; Do Nascimento, Antonio S; Dos Santos, José L; Cavichioli, Rodney R

2009-01-01

The causal agent of citrus variegated clorosis, Xylella fastidiosa, is transmitted by leafhoppers of the subfamily Cicadellinae, whose species vary regionally. The goal of this study was to identify potential vectors of this pathogen in citrus groves of Bahia North Coast, Brazil. The survey was done from March/2002 to February/2003 in three seven- to nine-year-old sweet orange (Citrus sinensis, Pêra variety) groves located in Rio Real, BA. Fifteen yellow sticky cards (8.5x11.5 cm) were installed 40 m apart in each grove, hanged at 1.5 m high on the upper north side of citrus canopies, and replaced fortnightly. A sweep net was periodically used to sample leafhoppers on herbaceous weeds inside the groves, by selecting five points at random and performing 30 sweeps in each point. A total of 1,360 specimens of 49 Auchenorrhyncha species were collected in this study, mostly in the family Cicadellidae (90.2%). The subfamily Cicadellinae, which includes the sharpshooter vectors of X. fastidiosa, showed the largest number of species (14) and specimens (84.8%). Acrogonia flagellata Young, A. citrina Marucci & Cavichioli, Homalodisca spottii Takiya, Cavichioli & McKamey and an unidentified Cicadellini (species 1) were the dominant species trapped on citrus canopies, while Hortensia similis (Walker) and Erythrogonia dubia (Medler) were dominant in the weedy vegetation. Among the Cicadellinae species already known as vectors of X. fastidiosa in citrus, only A. citrina, Bucephalogonia xanthophis (Berg) e Ferrariana trivittata (Signoret) were found. The two latter species were accidentally trapped by sweep net in the weedy vegetation.

Enhanced Acquisition Rates of 'Candidatus Liberibacter asiaticus' by the Asian Citrus Psyllid (Hemiptera: Liviidae) in the Presence of Vegetative Flush Growth in Citrus.

PubMed

Sétamou, Mamoudou; Alabi, Olufemi J; Kunta, Madhurababu; Jifon, John L; da Graça, John V

2016-10-01

The Asian citrus psyllid preferentially feeds and exclusively reproduces on young, newly emerged flush shoots of citrus. Asian citrus psyllid nymphs feed and complete their life stages on these flush shoots. Recent studies conducted under greenhouse conditions have shown that the transmission rates of 'Candidatus Liberibacter asiaticus' (CLas), the putative causal agent of huanglongbing disease of citrus, are enhanced when flush shoots are present. However, it is unclear if CLas acquisition by migrant adult Asian citrus psyllids is similarly enhanced. To address this knowledge gap, cohorts of Asian citrus psyllid adults were allowed 1-wk acquisition access period (AAP) on flushing and nonflushing shoots of qPCR-tested symptomatic (CLas+) and asymptomatic (CLas-) 10-yr-old sweet orange trees under field conditions. After the AAP, they were tested for CLas by qPCR. Progeny Asian citrus psyllid adults that emerged 4 wk post-AAP were similarly retrieved and tested. Eighty percent of flushing and 30% of nonflushing CLas+ trees produced infective Asian citrus psyllid adults, indicating that flush shoots have greater potential to be inoculum sources for CLas acquisition. Concomitantly, 21.1% and 6.0% infective adults were retrieved, respectively, from flushing and nonflushing CLas+ trees, indicating that Asian citrus psyllid adults acquire CLas more efficiently from flush shoots relative to mature shoots. In addition, 12.1% of infective Asian citrus psyllid adult progeny were obtained from 70% of flushing CLas+ trees. Significantly lower mean Ct values were also obtained from infective adults retrieved from flushing relative to nonflushing trees. The results underscore the role of flush shoots in CLas acquisition and the need to protect citrus trees from Asian citrus psyllid infestations during flush cycles. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email

Biological Indexing of Graft Transmissible Diseases of Citrus

USDA-ARS?s Scientific Manuscript database

Biological indexing for the detection of graft transmissible diseases of citrus is essential for maintaining a citrus certification program. Many of the graft transmissible diseases of citrus are harbored as latent infections in the scions, but when propagated on a susceptible rootstock that allow...

Salicylic acid-mediated establishment of the compatibility between Alternaria brassicicola and Brassica juncea is mitigated by abscisic acid in Sinapis alba.

PubMed

Mazumder, Mrinmoy; Das, Srirupa; Saha, Upala; Chatterjee, Madhuvanti; Bannerjee, Kaushik; Basu, Debabrata

2013-09-01

This work addresses the changes in the phytohormonal signature in the recognition of the necrotrophic fungal pathogen Alternaria brassicicola by susceptible Brassica juncea and resistant Sinapis alba. Although B. juncea, S. alba and Arabidopsis all belong to the same family, Brassicaceae, the phytohormonal response of susceptible B. juncea towards this pathogen is unique because the latter two species express non-host resistance. The differential expression of the PR1 gene and the increased level of salicylic acid (SA) indicated that an SA-mediated biotrophic mode of defence response was triggered in B. juncea upon challenge with the pathogen. Compared to B. juncea, resistant S. alba initiated enhanced abscisic acid (ABA) and jasmonic acid (JA) responses following challenge with this pathogen, as revealed by monitoring the expression of ABA-related genes along with the concentration of ABA and JA. Furthermore, these results were verified by the exogenous application of ABA on B. juncea leaves prior to challenge with A. brassicicola, which resulted in a delayed disease progression, followed by the inhibition of the pathogen-mediated increase in SA response and enhanced JA levels. Therefore, it seems that A. brassicicola is steering the defence response towards a biotrophic mode by mounting an SA response in susceptible B. juncea, whereas the enhanced ABA response of S. alba not only counteracts the SA response but also restores the necrotrophic mode of resistance by enhancing JA biosynthesis. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Site Plan & Site Section of Citrus Landscape (Showing Relationship ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Site Plan & Site Section of Citrus Landscape (Showing Relationship of Victoria Avenue to Citrus Groves) - Arlington Heights Citrus Landscape, Southwestern portion of city of Riverside, Riverside, Riverside County, CA

Expressed sequence enrichment for candidate gene analysis of citrus tristeza virus resistance.

PubMed

Bernet, G P; Bretó, M P; Asins, M J

2004-02-01

Several studies have reported markers linked to a putative resistance gene from Poncirus trifoliata ( Ctv-R) located at linkage group 4 that confers resistance against one of the most important citrus pathogens, citrus tristeza virus (CTV). To be successful in both marker-assisted selection and transformation experiments, its accurate mapping is needed. Several factors may affect its localization, among them two are considered here: the definition of resistance and the genetic background of progeny. Two progenies derived from P. trifoliata, by self-pollination and by crossing with sour orange ( Citrus aurantium), a citrus rootstock well-adapted to arid and semi-arid areas, were used for linkage group-4 marker enrichment. Two new methodologies were used to enrich this region with expressed sequences. The enrichment of group 4 resulted in the fusion of several C. aurantium linkage groups. The new one A(7+3+4) is now saturated with 48 markers including expressed sequences. Surprisingly, sour orange was as resistant to the CTV isolate tested as was P. trifoliata, and three hybrids that carry Ctv-R, as deduced from its flanking markers, are susceptible to CTV. The new linkage maps were used to map Ctv-R under the hypothesis of monogenic inheritance. Its position on linkage group 4 of P. trifoliata differs from the location previously reported in other progenies. The genetic analysis of virus-plant interaction in the family derived from C. aurantium after a CTV chronic infection showed the segregation of five types of interaction, which is not compatible with the hypothesis of a single gene controlling resistance. Two major issues are discussed: another type of genetic analysis of CTV resistance is needed to avoid the assumption of monogenic inheritance, and transferring Ctv-R from P. trifoliata to sour orange might not avoid the CTV decline of sweet orange trees.

ACP and Citrus: Plant Responses to Psyllid Feeding

USDA-ARS?s Scientific Manuscript database

Progress is reported on the Citrus Research Board funded project: 5300-150 Biomarkers for the detection of Liberibacter infection in citrus through H-NMR-based metabolomics. Proton nuclear magnetic resonance (H-NMR) was used to determine the effects of Asian citrus psyllid (ACP) feeding on leaf meta...

A comparison of plant species for rearing Asian citrus psyllid

USDA-ARS?s Scientific Manuscript database

Five plant genotypes were compared with respect to Asian citrus psyllid (ACP) reproduction potential: Bergera koenigii, Citrus aurantiifolia, C. macrophylla, C. taiwanica and Murraya paniculata. Asian citrus psyllid reproduction is dependent on young flush and thus Asian citrus psyllid production po...

Citrus tristeza virus-based RNAi in citrus plants induces gene silencing in Diaphorina citri, a phloem-sap sucking insect vector of citrus greening disease (Huanglongbing).

PubMed

Hajeri, Subhas; Killiny, Nabil; El-Mohtar, Choaa; Dawson, William O; Gowda, Siddarame

2014-04-20

A transient expression vector based on Citrus tristeza virus (CTV) is unusually stable. Because of its stability it is being considered for use in the field to control Huanglongbing (HLB), which is caused by Candidatus Liberibacter asiaticus (CLas) and vectored by Asian citrus psyllid, Diaphorina citri. In the absence of effective control strategies for CLas, emphasis has been on control of D. citri. Coincident cohabitation in phloem tissue by CLas, D. citri and CTV was exploited to develop a novel method to mitigate HLB through RNA interference (RNAi). Since CTV has three RNA silencing suppressors, it was not known if CTV-based vector could induce RNAi in citrus. Yet, expression of sequences targeting citrus phytoene desaturase gene by CTV-RNAi resulted in photo-bleaching phenotype. CTV-RNAi vector, engineered with truncated abnormal wing disc (Awd) gene of D. citri, induced altered Awd expression when silencing triggers ingested by feeding D. citri nymphs. Decreased Awd in nymphs resulted in malformed-wing phenotype in adults and increased adult mortality. This impaired ability of D. citri to fly would potentially limit the successful vectoring of CLas bacteria between citrus trees in the grove. CTV-RNAi vector would be relevant for fast-track screening of candidate sequences for RNAi-mediated pest control. Copyright © 2014. Published by Elsevier B.V.

Analysis of full-length sequences of two Citrus yellow mosaic badnavirus isolates infecting Citrus jambhiri (Rough Lemon) and Citrus sinensis L. Osbeck (Sweet Orange) from a nursery in India.

PubMed

Anthony Johnson, A M; Borah, B K; Sai Gopal, D V R; Dasgupta, I

2012-12-01

Citrus yellow mosaic badna virus (CMBV), a member of the Family Caulimoviridae, Genus Badnavirus is the causative agent of mosaic disease among Citrus species in southern India. Despite its reported prevalence in several citrus species, complete information on clear functional genomics or functional information of full-length genomes from all the CMBV isolates infecting citrus species are not available in publicly accessible databases. CMBV isolates from Rough Lemon and Sweet Orange collected from a nursery were cloned and sequenced. The analysis revealed high sequence homology of the two CMBV isolates with previously reported CMBV sequences implying that they represent new variants. Based on computational analysis of the predicted secondary structures, the possible functions of some CMBV proteins have been analyzed.

Chemical control of the Asian citrus psyllid and of huanglongbing disease in citrus.

PubMed

Boina, Dhana Raj; Bloomquist, Jeffrey R

2015-06-01

By 2014, huanglongbing (HLB), the most destructive disease of citrus, and its insect vector, the Asian citrus psyllid (ACP), Diaphorina citri (Kuwayama), became established in all major citrus-growing regions of the world, including the United States, with the exception of California. At present, application of insecticides is the most widely followed option for reducing ACP populations, while application of antibiotics for suppressing HLB disease/symptoms is being practiced in some citrus-growing regions. Application of insecticides during the dormant winter season, along with cultivation of HLB-free seedlings and early detection and removal of symptomatic and asymptomatic trees, has been very effective in managing ACP. Area-wide management of ACP by application of insecticides at low volume in large areas of citrus cultivation has been shown to be effective in managing HLB and reducing management costs. As insecticide resistance is a major problem in sustainable management of ACP, rotation/alternation of insecticides with different chemistries and modes of action needs to be followed. Besides control of the insect vector, use of antibiotics has temporarily suppressed the symptoms of HLB in diseased trees. Recent efforts to discover and screen existing as well as new compounds for their antibiotic and antimicrobial activities have identified some promising molecules for HLB control. There is an urgent need to find a sustainable solution to the HLB menace through chemical control of ACP populations and within HLB-infected trees through the judicious use of labeled insecticides (existing and novel chemistries) and antibiotics in area-wide management programs with due consideration to the insecticide resistance problem. © 2014 Society of Chemical Industry.

GC-MS analysis and antimicrobial activity determination of Citrus medica L. var proper leaf essential oil from South Sulawesi against skin pathogen microorganism

NASA Astrophysics Data System (ADS)

Aliyah; Himawan, A.; Rante, H.; Mufidah; Ningsih, D. R.

2017-11-01

A research about Citrus medica L. var sarcodactylis had been conducted and it showed a significant antimicrobial activity, thus drive our curiosity to investigate the other variety from the same species, Citrus medica L. var proper. This research focuses in chemical compound study and antimicrobial activity screening against Staphylococcus aureus, Propionibacterium acne, and Candida albicans of Citrus medica L. var Proper leaves’ essential oil. The essential oil is distillated from fresh leaves by hydrodestillation. The chemical compound was analysed using GC-MS instrument while the antimicrobial activity was tested using disk diffusion method. The results showed that the major component of the essential oil was Z-citral, citral and limonene compounds. The antimicrobial activity test results against the test microorganism are 9.15±0.15 mm, 11.15±1.3 mm and 8.02±0.48 mm, consecutively, for Staphylococcus aureus, Propionibacterium acne and Candida albicans.

Determination of polymethoxylated flavones in peels of selected Jamaican and Mexican citrus (Citrus spp.) cultivars by high-performance liquid chromatography.

PubMed

Green, Curtis O; Wheatley, Andrew O; Osagie, Anthony U; St A Morrison, Errol Y; Asemota, Helen N

2007-01-01

The concentrations of the polymethoxylated flavones (PMFs) in peels of selected citrus cultivars grown in Jamaica and Mexico were determined. The PMFs were extracted from sun-dried citrus peels with reagent-grade methanol. Analyses were carried out by reverse-phase HPLC and UV detection. The column used was a C(18) 5 microm (150 x 4.6 mm) Discovery column. Elution was in the gradient mode, using a ternary mobile phase. The results showed that all the citrus cultivars used contained at least three of the six major PMFs quantified. Ortanique peel contained the highest quantity of PMFs (34,393 +/- 272 ppm), followed by tangerine (28,389 +/- 343 ppm) and Mexican sweet orange (sample 1; 21,627 +/- 494 ppm). The major PMFs, i.e. sinensetin, nobiletin, tangeretin, heptamethoxyflavone, tetramethylscutellarein and hexamethyl-o-quercetagetin, present in the peels of 20 citrus cultivars, was quantified. The results were compared with those of Florida citrus peels. A large amount of citrus peels and byproducts are produced in the Caribbean which could provide a cheap and convenient source of PMFs. Copyright 2006 John Wiley & Sons, Ltd.

Development of a simple and rapid reverse transcription-loop mediated isothermal amplification (RT-LAMP) assay for sensitive detection of Citrus tristeza virus.

PubMed

Warghane, Ashish; Misra, Pragati; Bhose, Sumit; Biswas, Kajal Kumar; Sharma, Ashwani Kumar; Reddy, M Krishna; Ghosh, Dilip Kumar

2017-12-01

Tristeza is a devastating disease of citrus and reported to be present in almost all countries where it is cultivated as a commercial crop. The etiological agent of this disease is Citrus tristeza virus (CTV), a member of the genus Closterovirus with in the family Closteroviridae. The pathogen is restricted to the phloem tissue of the infected citrus plant and has a monopartite ss (+) RNA genome of ∼20kb size. Till date, there is no effective control measure available for this virus. Management of tristeza depends on destruction of CTV infected field plants, production of virus-free planting material for new orchard establishment and controlling viruliferous aphid vectors responsible for field spread of the pathogen. Availability of rapid diagnostic assay is essential for rapid and efficient detection of the pathogen. In the present investigation, RT-LAMP (reverse transcription-loop mediated isothermal amplification), a highly sensitive, robust and low cost assay has been developed for rapid detection of CTV in infected citrus plant samples. Based on conserved nucleotide sequences available in GenBank and specific to p25 gene (major coat protein gene) of predominant CTV isolates of India, four primer sets (CTV-F3, CTV-B3, CTV-FIP and CTV-BIP) ware designed and custom synthesized. The amplified LAMP products obtained after maintaining isothermal condition of 65°C for 60min duration could be visible easily with necked eyes in presence of SYBR Green I (100X). Subsequently, LAMP products were verified by electrophoresis run in 1.5% agarose gel. The RT-LAMP results obtained with known CTV isolates maintained in screen house of CCRI, Nagpur were validated using field samples and thereafter it was further confirmed by conventional RT-PCR (reveres transcription-polymerase chain reaction) assay. The sensitivity of CTV-RT-LAMP protocol standardized in the present study was 100 times more than conventional one step RT-PCR assay. It also has maximum detection limit up to 0

Herbivory by the insect diaphorina citri induces greater change in citrus plant volatile profile than does infection by the bacterium, Candidatus Liberibacter asiaticus

PubMed Central

Hijaz, Faraj; El-Shesheny, Ibrahim; Killiny, Nabil

2013-01-01

The volatile organic compound (VOC) profile in plant leaves often changes after biotic and abiotic stresses. Monitoring changes in VOCs in plant leaves could provide valuable information about multitrophic interactions. In the current study, we investigated the effect of Asian citrus psyllid (ACP) infestation, citrus greening pathogen (Candidatus Liberibacter asiaticus [CLas]) infection, and simultaneous attack by ACP and CLas on the VOC content of citrus leaves. Leaf volatiles were extracted using hexane and analyzed with gas chromatography-mass spectrometry (GC-MS). Although ACP is a phloem-sucking insect that causes minimal damage to plant tissues, the relative amount of 21 out of the 27 VOCs increased 2- to 10-fold in ACP-infested plants. The relative amount of d-limonene, β-phelandrene, citronellal, and undecanal were increased 4- to 20- fold in CLas-infected plants. A principle component analysis (PCA) and cluster analysis (CA) showed that VOC patterns of ACP-infested and CLas-infected plants were different from each other and were also different from the controls, while the VOC pattern of double-attacked plants was more like that of the controls than that of ACP-infested or CLas-infected plants. VOC amounts from leaves were compromised when plants were attacked by ACP and CLas. The results of this study showed that a simple direct extraction of citrus leaf volatiles could be successfully used to discriminate between healthy and CLas-infected plants. Information about the effects of insect and pathogen attack on the VOC content profile of plants might contribute to a better understanding of biotic stress. PMID:23857364

Comparison of evapotranspiration rates for flatwoods and ridge citrus

USGS Publications Warehouse

Jia, X.; Swancar, A.; Jacobs, J.M.; Dukes, M.D.; Morgan, K.

2007-01-01

Florida citrus groves are typically grown in two regions of the state: flatwoods and ridge. The southern flatwoods citrus area has poorly drained fine textured sands with low organic matter in the shallow root zone. Ridge citrus is located in the northern ridge citrus zone and has fine to coarse textured sands with low water-holding capacity. Two commercial citrus groves, selected from each region, were studied from 15 July 2004 to 14 July 2005. The flatwoods citrus (FC) grove had a grass cover and used drainage ditches to remove excess water from the root zone. The ridge citrus (RC) grove had a bare soil surface with weeds periodically eliminated by tillage. Citrus crop evapotranspiration (ETc) rates at the two citrus groves were measured by the eddy correlation method, and components in the energy balance were also examined and compared. The study period had higher than average rainfall, and as a result, the two locations had similar annual ETc rates (1069 and 1044 mm for RC and FC, respectively). The ETc rates were 59% (RC) and 47% (FC) of the rainfall amounts during the study period. The annual reference crop evapotranspiration (ETo) rates were 1180 mm for RC and 1419 mm for FC, estimated using the standardized reference evapotranspiration equation. The citrus crop coefficients (Kc, ratio of ETc to ET o) were different between the two locations because of differences in latitude, ground cover, and rainfall amounts. The Kc values ranged from 0.70 between December and March to 1.05 between July and November for RC, and from 0.65 between November and May to 0.85 between June and October for FC. The results are consistent with other Kc values reported from field studies on citrus in both Florida and elsewhere using these and alternate methods.

Changes in Anthocyanin Production during Domestication of Citrus1[OPEN

PubMed Central

Garcia-Lor, Andrés; Licciardello, Concetta; Las Casas, Giuseppina; Ramadugu, Chandrika; Krueger, Robert; Fanciullino, Anne-Laure; Froelicher, Yann

2017-01-01

Mandarin (Citrus reticulata), citron (Citrus medica), and pummelo (Citrus maxima) are important species of the genus Citrus and parents of the interspecific hybrids that constitute the most familiar commercial varieties of Citrus: sweet orange, sour orange, clementine, lemon, lime, and grapefruit. Citron produces anthocyanins in its young leaves and flowers, as do species in genera closely related to Citrus, but mandarins do not, and pummelo varieties that produce anthocyanins have not been reported. We investigated the activity of the Ruby gene, which encodes a MYB transcription factor controlling anthocyanin biosynthesis, in different accessions of a range of Citrus species and in domesticated cultivars. A white mutant of lemon lacks functional alleles of Ruby, demonstrating that Ruby plays an essential role in anthocyanin production in Citrus. Almost all the natural variation in pigmentation by anthocyanins in Citrus species can be explained by differences in activity of the Ruby gene, caused by point mutations and deletions and insertions of transposable elements. Comparison of the allelic constitution of Ruby in different species and cultivars also helps to clarify many of the taxonomic relationships in different species of Citrus, confirms the derivation of commercial varieties during domestication, elucidates the relationships within the subgenus Papeda, and allows a new genetic classification of mandarins. PMID:28196843

Field validation of a system for autodissemination of an entomopathogenic fungus, Isaria fumosorosea, to control the Asian citrus psyllid on residential citrus

USDA-ARS?s Scientific Manuscript database

The citrus industries of California and Texas share a pressing problem with the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae) and huanglongbing (HLB) spreading in residential citrus near commercial groves. Insecticidal treatment of residential trees for the psyllid is problem...

In Silico Identification of Mimicking Molecules as Defense Inducers Triggering Jasmonic Acid Mediated Immunity against Alternaria Blight Disease in Brassica Species

PubMed Central

Pathak, Rajesh K.; Baunthiyal, Mamta; Shukla, Rohit; Pandey, Dinesh; Taj, Gohar; Kumar, Anil

2017-01-01

Alternaria brassicae and Alternaria brassicicola are two major phytopathogenic fungi which cause Alternaria blight, a recalcitrant disease on Brassica crops throughout the world, which is highly destructive and responsible for significant yield losses. Since no resistant source is available against Alternaria blight, therefore, efforts have been made in the present study to identify defense inducer molecules which can induce jasmonic acid (JA) mediated defense against the disease. It is believed that JA triggered defense response will prevent necrotrophic mode of colonization of Alternaria brassicae fungus. The JA receptor, COI1 is one of the potential targets for triggering JA mediated immunity through interaction with JA signal. In the present study, few mimicking compounds more efficient than naturally occurring JA in terms of interaction with COI1 were identified through virtual screening and molecular dynamics simulation studies. A high quality structural model of COI1 was developed using the protein sequence of Brassica rapa. This was followed by virtual screening of 767 analogs of JA from ZINC database for interaction with COI1. Two analogs viz. ZINC27640214 and ZINC43772052 showed more binding affinity with COI1 as compared to naturally occurring JA. Molecular dynamics simulation of COI1 and COI1-JA complex, as well as best screened interacting structural analogs of JA with COI1 was done for 50 ns to validate the stability of system. It was found that ZINC27640214 possesses efficient, stable, and good cell permeability properties. Based on the obtained results and its physicochemical properties, it is capable of mimicking JA signaling and may be used as defense inducers for triggering JA mediated resistance against Alternaria blight, only after further validation through field trials. PMID:28487711

Antibacterial effect of citrus press-cakes dried by high speed and far-infrared radiation drying methods

PubMed Central

Samarakoon, Kalpa; Senevirathne, Mahinda; Lee, Won-Woo; Kim, Young-Tae; Kim, Jae-Il; Oh, Myung-Cheol

2012-01-01

In this study, the antibacterial effect was evaluated to determine the benefits of high speed drying (HSD) and far-infrared radiation drying (FIR) compared to the freeze drying (FD) method. Citrus press-cakes (CPCs) are released as a by-product in the citrus processing industry. Previous studies have shown that the HSD and FIR drying methods are much more economical for drying time and mass drying than those of FD, even though FD is the most qualified drying method. The disk diffusion assay was conducted, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined with methanol extracts of the dried CPCs against 11 fish and five food-related pathogenic bacteria. The disk diffusion results indicated that the CPCs dried by HSD, FIR, and FD prevented growth of all tested bacteria almost identically. The MIC and MBC results showed a range from 0.5-8.0 mg/mL and 1.0-16.0 mg/mL respectively. Scanning electron microscopy indicated that the extracts changed the morphology of the bacteria cell wall, leading to destruction. These results suggest that CPCs dried by HSD and FIR showed strong antibacterial activity against pathogenic bacteria and are more useful drying methods than that of the classic FD method in CPCs utilization. PMID:22808341

Monoclonal antibody-based serological methods for detecting Citrus tristeza virus in citrus groves.

PubMed

Liu, Zhen; Chen, Zhe; Hong, Jian; Wang, Xuefeng; Zhou, Changyong; Zhou, Xueping; Wu, Jianxiang

2016-08-01

Citrus tristeza virus (CTV) is one of the most economically important citrus viruses and harms the citrus industry worldwide. To develop reliable and effective serological detection assays of CTV, the major capsid protein (CP) gene of CTV was expressed in Escherichia coli BL21 (DE3) using the expression vector pET-28a and purified through Ni+-NTA affinity chromatography. The recombinant protein was used to immunize BALB/c mice. Four hybridoma cell lines (14B10, 14H11, 20D5, and 20G12) secreting monoclonal antibodies (MAbs) against CTV were obtained through conventional hybridoma technology. The titers of MAb-containing ascitic fluids secreted by the four hybridoma lines ranged from 10(-6) to 10(-7) in indirect enzyme-linked immunosorbent assay (ELISA). Western blots showed that all four MAbs could specifically react with CTV CP. Using the prepared MAbs, dot-ELISA, Tissue print-ELISA, and triple antibody sandwich (TAS)-ELISA were developed to detect CTV in tree nurseries and epidemiological studies. The developed dot-ELISA and TAS-ELISA methods could detect CTV in crude extracts of infected citrus leaves with dilutions of 1:2560 and 1:10, 240 (w/v, g/mL), respectively. Tissue print-ELISA was particularly useful for large-scale field sample detection, mainly owing to its simplicity and lack of sample preparation requirements. The field survey revealed that CTV is prevalent on citrus trees in the Chongqing Municipality, Jiangxi Province, and Zhejiang Province of China. The coincidence rate of serological and RT-PCR test results reached more than 99.5%. The prepared MAbs against CTV and established sensitive and specific serological assays have a significant role in the detection and prevention and control of CTV in our country.

Imidacloprid soil movement under micro-sprinkler irrigation and soil-drench applications to control Asian citrus psyllid (ACP) and citrus leafminer (CLM).

PubMed

Fletcher, Evelyn; Morgan, Kelly T; Qureshi, Jawwad A; Leiva, Jorge A; Nkedi-Kizza, Peter

2018-01-01

Imidacloprid (IM) is used to control the Asian Citrus Psyllid (ACP) and citrus leafminer (CLM), which are related to the spread of huanglongbing (HLB or citrus greening) and citrus canker diseases, respectively. In Florida citrus, imidacloprid is mainly soil-drenched around the trees for proper root uptake and translocation into plant canopy to impact ACP and CLM. The objective of this study was to determine the effect of imidacloprid rate, and irrigate amount on concentration of imidacloprid in the soil following drench application to citrus trees in three age classes. The plots were established at the Southwest Florida Research and Education Center, Immokalee, using a randomized complete-block design for three age classes of trees: one-year-old trees (B1), three to five-year-old trees (B2), and eight-year-old trees (B3). The treatments were a combination of two rates each of imidacloprid (1D, 2D) and micro-sprinkling irrigation (1I, 2I). Imidacloprid and bromide (Br-) used as tracer were applied simultaneously. Soil moisture and concentrations of imidacloprid and Br were monitored using soil cores from hand held augers. Soil moisture content (θV) did not differ under two irrigation rates at any given observation day or depth, except following heavy rainfall events. Br- was lost from the observation depths (0-45 cm) about two weeks after soil-drench. Contrarily, imidacloprid persisted for a much longer time (4-8 weeks) at all soil depths, regardless of treatment combinations. The higher retardation of imidacloprid was related to the predominantly unsaturated conditions of the soil (which in turn reduced soil hydraulic conductivities by orders of magnitude), the imidacloprid sorption on soil organic matter, and the citrus root uptake. Findings of this study are important for citrus growers coping with the citrus greening and citrus canker diseases because they suggest that imidacloprid soil drenches can still be an effective control measure of ACP and CLM, and the

Imidacloprid soil movement under micro-sprinkler irrigation and soil-drench applications to control Asian citrus psyllid (ACP) and citrus leafminer (CLM)

PubMed Central

Fletcher, Evelyn; Qureshi, Jawwad A.; Leiva, Jorge A.; Nkedi-Kizza, Peter

2018-01-01

Imidacloprid (IM) is used to control the Asian Citrus Psyllid (ACP) and citrus leafminer (CLM), which are related to the spread of huanglongbing (HLB or citrus greening) and citrus canker diseases, respectively. In Florida citrus, imidacloprid is mainly soil-drenched around the trees for proper root uptake and translocation into plant canopy to impact ACP and CLM. The objective of this study was to determine the effect of imidacloprid rate, and irrigate amount on concentration of imidacloprid in the soil following drench application to citrus trees in three age classes. The plots were established at the Southwest Florida Research and Education Center, Immokalee, using a randomized complete-block design for three age classes of trees: one-year-old trees (B1), three to five-year-old trees (B2), and eight-year-old trees (B3). The treatments were a combination of two rates each of imidacloprid (1D, 2D) and micro-sprinkling irrigation (1I, 2I). Imidacloprid and bromide (Br-) used as tracer were applied simultaneously. Soil moisture and concentrations of imidacloprid and Br were monitored using soil cores from hand held augers. Soil moisture content (θV) did not differ under two irrigation rates at any given observation day or depth, except following heavy rainfall events. Br- was lost from the observation depths (0–45 cm) about two weeks after soil-drench. Contrarily, imidacloprid persisted for a much longer time (4–8 weeks) at all soil depths, regardless of treatment combinations. The higher retardation of imidacloprid was related to the predominantly unsaturated conditions of the soil (which in turn reduced soil hydraulic conductivities by orders of magnitude), the imidacloprid sorption on soil organic matter, and the citrus root uptake. Findings of this study are important for citrus growers coping with the citrus greening and citrus canker diseases because they suggest that imidacloprid soil drenches can still be an effective control measure of ACP and CLM, and

Citrus Flavonoids as Regulators of Lipoprotein Metabolism and Atherosclerosis.

PubMed

Mulvihill, Erin E; Burke, Amy C; Huff, Murray W

2016-07-17

Citrus flavonoids are polyphenolic compounds with significant biological properties. This review summarizes recent advances in understanding the ability of citrus flavonoids to modulate lipid metabolism, other metabolic parameters related to the metabolic syndrome, and atherosclerosis. Citrus flavonoids, including naringenin, hesperitin, nobiletin, and tangeretin, have emerged as potential therapeutics for the treatment of metabolic dysregulation. Epidemiological studies reveal an association between the intake of citrus flavonoid-containing foods and a decreased incidence of cardiovascular disease. Studies in cell culture and animal models, as well as a limited number of clinical studies, reveal the lipid-lowering, insulin-sensitizing, antihypertensive, and anti-inflammatory properties of citrus flavonoids. In animal models, supplementation of rodent diets with citrus flavonoids prevents hepatic steatosis, dyslipidemia, and insulin resistance primarily through inhibition of hepatic fatty acid synthesis and increased fatty acid oxidation. Citrus flavonoids blunt the inflammatory response in metabolically important tissues including liver, adipose, kidney, and the aorta. The mechanisms underlying flavonoid-induced metabolic regulation have not been completely established, although several potential targets have been identified. In mouse models, citrus flavonoids show marked suppression of atherogenesis through improved metabolic parameters as well as through direct impact on the vessel wall. Recent studies support a role for citrus flavonoids in the treatment of dyslipidemia, insulin resistance, hepatic steatosis, obesity, and atherosclerosis. Larger human studies examining dose, bioavailability, efficacy, and safety are required to promote the development of these promising therapeutic agents.

Rapid cyling plant breeding in citrus

USDA-ARS?s Scientific Manuscript database

Resistance or tolerance to huanglongbing (HLB) and other important traits have been identified in several citrus types and relatives and associated markers should be identified soon. What is urgently needed in addition is an accelerated strategy for citrus variety breeding. Identification and use of...

Effect of genotype and environment on citrus juice carotenoid content.

PubMed

Dhuique-Mayer, Claudie; Fanciullino, Anne-Laure; Dubois, Cecile; Ollitrault, Patrick

2009-10-14

A selection of orange and mandarin varieties belonging to the same Citrus accession and cultivated in Mediterranean (Corsica), subtropical (New Caledonia), and tropical areas (principally Tahiti) were studied to assess the effect of genotype and environmental conditions on citrus juice carotenoid content. Juices from three sweet orange cultivars, that is, Pera, Sanguinelli, and Valencia ( Citrus sinensis (L.) Osbeck), and two mandarin species ( Citrus deliciosa Ten and Citrus clementina Hort. ex Tan), were analyzed by HPLC using a C(30) column. Annual carotenoid content variations in Corsican fruits were evaluated. They were found to be very limited compared to variations due to varietal influences. The statistical analysis (PCA, dissimilarity tree) results based on the different carotenoid compounds showed that citrus juice from Corsica had a higher carotenoid content than citrus juices from tropical origins. The tropical citrus juices were clearly differentiated from citrus juices from Corsica, and close correlations were obtained between beta-cryptoxanthin and phytoene (r = 0.931) and beta-carotene and phytoene (r = 0.918). More broadly, Mediterranean conditions amplified interspecific differentiation, especially by increasing the beta-cryptoxanthin and cis-violaxanthin content in oranges and beta-carotene and phytoene-phytofluene content in mandarins. Thus, at a quantitative level, environmental conditions also had a major role in determining the levels of carotenoids of nutritional interest, such as the main provitamin A carotenoids in citrus juice (beta-cryptoxanthin and beta-carotene).

Compatibility of Isaria fumosorosea (Hypocreales: Cordycipitaceae) Blastospores with Agricultural Chemicals Used for Management of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae)

PubMed Central

Avery, Pasco B.; Pick, David A.; Aristizábal, Luis F.; Kerrigan, James; Powell, Charles A.; Rogers, Michael E.; Arthurs, Steven P.

2013-01-01

Biorational insecticides are being increasingly emphasized for inclusion in integrated pest management programs for invasive insects. The entomopathogenic fungus, Isaria fumosorosea, can be used to help manage the Asian citrus psyllid with minimal impact on beneficial arthropods, but its effectiveness may be compromised by agrochemicals used to control concurrent arthropod pests and diseases. We evaluated the compatibility of I. fumosorosea blastospores with a range of spray oils and copper-based fungicides registered for use in citrus groves. Results of laboratory and greenhouse tests showed a range of responses of the fungus to the different materials, including compatibility and incompatibility. Overall, I. fumosorosea growth in vitro was reduced least by petroleum-based materials and most by botanical oils and borax, and some of the copper-based fungicides, suggesting that tank mixing of I. fumosorosea with these latter products should be avoided. However, equivalent negative effects of test materials on fungal pathogenicity were not always observed in tests with adult psyllids. We hypothesize that some oils enhanced adherence of blastospores to the insect cuticle, overcoming negative impacts on germination. Our data show that care should be taken in selecting appropriate agrochemicals for tank-mixing with commercial formulations of entomopathogenic fungi for management of citrus pests. The prospects of using I. fumosorosea for managing the invasive Asian citrus psyllid and other citrus pests are discussed. PMID:26462531

Temperature Studies with the Asian Citrus Psyllid, Diaphorina citri: Cold Hardiness and Temperature Thresholds for Oviposition

PubMed Central

Hall, David G.; Wenninger, Erik J.; Hentz, Matthew G.

2011-01-01

This study was conducted to obtain information on the cold hardiness of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), in Florida and to assess upper and lower temperature thresholds for oviposition. The psyllid is an important pest in citrus because it transmits the bacterial pathogens responsible for citrus greening disease, Huanglongbing, considered the most serious citrus disease worldwide. D. citri was first found in Florida during 1998, and the disease was discovered during 2005. Little was known regarding cold hardiness of D. citri, but Florida citrus is occasionally subjected to notable freeze events. Temperature and duration were each significant sources of variation in percent mortality of D. citri subjected to freeze events. Relatively large percentages of adults and nymphs survived after being exposed for several hours to temperatures as low as -5 to -6° C. Relatively large percentages of eggs hatched after being exposed for several hours to temperatures as low as -8° C. Research results indicated that adult D. citri become cold acclimated during the winter through exposure to cooler winter temperatures. There was no evidence that eggs became cold acclimated during winter. Cold acclimation in nymphs was not investigated. Research with adult D. citri from laboratory and greenhouse colonies revealed that mild to moderate freeze events were usually nonlethal to the D. citri irrespective of whether they were cold acclimated or not. Upper and lower temperature thresholds for oviposition were investigated because such information may be valuable in explaining the geographic distribution and potential spread of the pest from Florida as well as how cooler winter temperatures might limit population growth. The estimated lower and upper thresholds for oviposition were 16.0 and 41.6° C, respectively; the estimated temperature of peak oviposition over a 48 h period was 29.6° C. PMID:21870969

Temperature studies with the Asian citrus psyllid, Diaphorina citri: cold hardiness and temperature thresholds for oviposition.

PubMed

Hall, David G; Wenninger, Erik J; Hentz, Matthew G

2011-01-01

This study was conducted to obtain information on the cold hardiness of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), in Florida and to assess upper and lower temperature thresholds for oviposition. The psyllid is an important pest in citrus because it transmits the bacterial pathogens responsible for citrus greening disease, Huanglongbing, considered the most serious citrus disease worldwide. D. citri was first found in Florida during 1998, and the disease was discovered during 2005. Little was known regarding cold hardiness of D. citri, but Florida citrus is occasionally subjected to notable freeze events. Temperature and duration were each significant sources of variation in percent mortality of D. citri subjected to freeze events. Relatively large percentages of adults and nymphs survived after being exposed for several hours to temperatures as low as -5 to -6 °C. Relatively large percentages of eggs hatched after being exposed for several hours to temperatures as low as -8 °C. Research results indicated that adult D. citri become cold acclimated during the winter through exposure to cooler winter temperatures. There was no evidence that eggs became cold acclimated during winter. Cold acclimation in nymphs was not investigated. Research with adult D. citri from laboratory and greenhouse colonies revealed that mild to moderate freeze events were usually nonlethal to the D. citri irrespective of whether they were cold acclimated or not. Upper and lower temperature thresholds for oviposition were investigated because such information may be valuable in explaining the geographic distribution and potential spread of the pest from Florida as well as how cooler winter temperatures might limit population growth. The estimated lower and upper thresholds for oviposition were 16.0 and 41.6 °C, respectively; the estimated temperature of peak oviposition over a 48 h period was 29.6 °C.

Photographic Remote Sensing of Sick Citrus Trees

NASA Technical Reports Server (NTRS)

Gausman, H. W.

1971-01-01

Remote sensing with infrared color aerial photography (Kodak Ektachrome Infrared Aero 8443 film) for detecting citrus tree anomalies is described. Illustrations and discussions are given for detecting nutrient toxicity symptoms, for detecting foot rot and sooty mold fungal diseases, and for distinguishing among citrus species. Also, the influence of internal leaf structure on light reflectance, transmittance, and absorptance are considered; and physiological and environmental factors that affect citrus leaf light reflectance are reviewed briefly and illustrated.

Deep Sequencing Analysis of RNAs from Citrus Plants Grown in a Citrus Sudden Death-Affected Area Reveals Diverse Known and Putative Novel Viruses.

PubMed

Matsumura, Emilyn E; Coletta-Filho, Helvecio D; Nouri, Shahideh; Falk, Bryce W; Nerva, Luca; Oliveira, Tiago S; Dorta, Silvia O; Machado, Marcos A

2017-04-24

Citrus sudden death (CSD) has caused the death of approximately four million orange trees in a very important citrus region in Brazil. Although its etiology is still not completely clear, symptoms and distribution of affected plants indicate a viral disease. In a search for viruses associated with CSD, we have performed a comparative high-throughput sequencing analysis of the transcriptome and small RNAs from CSD-symptomatic and -asymptomatic plants using the Illumina platform. The data revealed mixed infections that included Citrus tristeza virus (CTV) as the most predominant virus, followed by the Citrus sudden death-associated virus (CSDaV), Citrus endogenous pararetrovirus (CitPRV) and two putative novel viruses tentatively named Citrus jingmen-like virus (CJLV), and Citrus virga-like virus (CVLV). The deep sequencing analyses were sensitive enough to differentiate two genotypes of both viruses previously associated with CSD-affected plants: CTV and CSDaV. Our data also showed a putative association of the CSD-symptomatic plants with a specific CSDaV genotype and a likely association with CitPRV as well, whereas the two putative novel viruses showed to be more associated with CSD-asymptomatic plants. This is the first high-throughput sequencing-based study of the viral sequences present in CSD-affected citrus plants, and generated valuable information for further CSD studies.

Laser diagnostic technology for early detection of pathogen infestation in orange fruits

NASA Astrophysics Data System (ADS)

Giubileo, Gianfranco; Lai, Antonella; Piccinelli, Delinda; Puiu, Adriana

2010-11-01

Due to an increased expectation of food products that respect high quality and safety standards, there is a need for the growth of accurate, fast, objective and non-destructive technologies for quality determination of food and agricultural products. For this purpose, a diagnostic system based on laser photoacoustic spectroscopy (LPAS) was developed at ENEA Frascati Molecular Spectroscopy Laboratory (Italy). In the design of the photoacoustic detector, particular emphasis was placed in attaining a high sensitivity in detecting ethylene (ET) down to sub-parts per billion level (minimum detectable concentration 0.2 ppb). This was required due to the necessity to monitor and follow up ET production at a single fruit scale. ET is normally synthesised in very low amounts by healthy citrus fruits; however stress conditions such as pathogen attack may induce a substantial increase in the synthesised ET. In the present paper, the comparison between the ET emitted by healthy oranges ( Citrus sinensis L. Osbeck) cv Navel and by Phytophthora citrophthora infested Navel orange fruits are reported. The obtained results show a well evident increase in ET emission from the infested fruit with respect to the healthy one, even 24 h after the inoculation with the pathogen; at that time the tissue necrosis was not yet visible, and the fruit was also not yet damaged. The possibility to perform a real time non-destructive detection of ET traces makes the LPAS a powerful tool for monitoring the healthy state of the citrus fruits.

Homologues of CsLOB1 in citrus function as disease susceptibility genes in citrus canker.

PubMed

Zhang, Junli; Huguet-Tapia, Jose Carlos; Hu, Yang; Jones, Jeffrey; Wang, Nian; Liu, Sanzhen; White, Frank F

2017-08-01

The lateral organ boundary domain (LBD) genes encode a group of plant-specific proteins that function as transcription factors in the regulation of plant growth and development. Citrus sinensis lateral organ boundary 1 (CsLOB1) is a member of the LBD family and functions as a disease susceptibility gene in citrus bacterial canker (CBC). Thirty-four LBD members have been identified from the Citrus sinensis genome. We assessed the potential for additional members of LBD genes in citrus to function as surrogates for CsLOB1 in CBC, and compared host gene expression on induction of different LBD genes. Using custom-designed transcription activator-like (TAL) effectors, two members of the same clade as CsLOB1, named CsLOB2 and CsLOB3, were found to be capable of functioning similarly to CsLOB1 in CBC. RNA sequencing and quantitative reverse transcription-polymerase chain reaction analyses revealed a set of cell wall metabolic genes that are associated with CsLOB1, CsLOB2 and CsLOB3 expression and may represent downstream genes involved in CBC. © 2016 BSPP AND JOHN WILEY & SONS LTD.

Effect of Nigerian citrus (Citrus sinensis Osbeck) honey on ethanol metabolism.

PubMed

Onyesom, I

2004-12-01

The effect of Nigerian citrus (Citrus sinensis Osbeck) honey on ethanol metabolism was tested using 45 consenting individuals in apparent good health and between the ages of 25 and 35 years. The subjects were moderate social drinkers matched in terms of body weight and build. The results obtained showed that on average, honey significantly (p < 0.05) increased the blood ethanol clearance rate by 68% and decreased the intoxication period by 43%, but insignificantly (p > 0.05) reduced the degree of intoxication by 9%. Honey could be a promising anti-intoxicating agent, but its long-term biochemical evaluation, possibly as a complement in the management of alcohol intoxication, deserves further study.

Effect of citrus byproducts on survival of O157:H7 and non-O157 Escherichia coli serogroups within in vitro bovine ruminal microbial fermentations

USDA-ARS?s Scientific Manuscript database

Citrus by-products contain essential oils that possess antimicrobial activities that can exert damage to the cell wall of gram-negative bacteria. This alteration to gram-negative microbes has resulted in CBP being investigated as a potential pre-harvest pathogen intervention strategy to reduce Shig...

Alternaria sp. MG1, a resveratrol-producing fungus: isolation, identification, and optimal cultivation conditions for resveratrol production.

PubMed

Shi, Junling; Zeng, Qin; Liu, Yanlin; Pan, Zhongli

2012-07-01

Due to its potential in preventing or slowing the occurrence of many diseases, resveratrol (3,5,4'-trihydroxystilbene) has attracted great research interest. The objective of this study was to identify microorganisms from selected plants that produce resveratrol and to optimize the conditions for resveratrol production. Endophytes from Merlot wine grapes (Vitis vinifera L. cv. Merlot), wild Vitis (Vitis quinquangularis Rehd.), and Japanese knotweed (Polygonum cuspidatum Siebold & Zucc.) were isolated, and their abilities to produce resveratrol were evaluated. A total of 65 isolates were obtained and 21 produced resveratrol (6-123 μg/L) in liquid culture. The resveratrol-producing isolates belonged to seven genera, Botryosphaeria, Penicillium, Cephalosporium, Aspergillus, Geotrichum, Mucor, and Alternaria. The resveratrol-producing capability decreased or was completely lost in most isolates after three rounds of subculture. It was found that only the strain Alternaria sp. MG1 (isolated from cob of Merlot using GA1 medium) had stable and high resveratrol-producing capability in all subcultures. During liquid cultivation of Alternaria sp. MG1 in potato dextrose medium, the synthesis of resveratrol began on the first day, increased to peak levels on day 7, and then decreased sharply thereafter. Cell growth increased during cultivation and reached a stable and high level of biomass after 5 days. The best fermentation conditions for resveratrol production in liquid cultures of Alternaria sp. MG1 were an inoculum size of 6 %, a medium volume of 125 mL in a 250-mL flask, a rotation speed of 101 rpm, and a temperature of 27 °C.

Discovering Novel Alternaria solani Succinate Dehydrogenase Inhibitors by In Silico Modeling and Virtual Screening Strategies to Combat Early Blight

NASA Astrophysics Data System (ADS)

Iftikhar, Sehrish; Shahid, Ahmad A.; Halim, Sobia A.; Wolters, Pieter J.; Vleeshouwers, Vivianne G. A. A.; Khan, Ajmal; Al-Harrasi, Ahmed; Ahmad, Shahbaz

2017-11-01

Alternaria blight is an important foliage disease caused by Alternaria solani. The enzyme Succinate dehydrogenase (SDH) is a potential drug target because of its role in tricarboxylic acid cycle. Hence targeting Alternaria solani SDH enzyme could be efficient tool to design novel fungicides against A. solani. We employed computational methodologies to design new SDH inhibitors using homology modeling; pharmacophore modeling and structure based virtual screening protocol. The three dimensional SDH model showed good stereo-chemical and structural properties. Based on virtual screening results twelve commercially available compounds were purchased and tested in vitro and in vivo. The compounds were found to inhibit mycelial growth of A. solani. Moreover in vitro trials showed that inhibitory effects were enhanced with increase in concentrations. Similarly increased disease control was observed in pre-treated potato tubers. Hence the applied in silico strategy led us to identify new and novel fungicides.

Discovering Novel Alternaria solani Succinate Dehydrogenase Inhibitors by in Silico Modeling and Virtual Screening Strategies to Combat Early Blight

PubMed Central

Iftikhar, Sehrish; Shahid, Ahmad A.; Halim, Sobia A.; Wolters, Pieter J.; Vleeshouwers, Vivianne G. A. A.; Khan, Ajmal; Al-Harrasi, Ahmed; Ahmad, Shahbaz

2017-01-01

Alternaria blight is an important foliage disease caused by Alternaria solani. The enzyme Succinate dehydrogenase (SDH) is a potential drug target because of its role in tricarboxylic acid cycle. Hence targeting Alternaria solani SDH enzyme could be efficient tool to design novel fungicides against A. solani. We employed computational methodologies to design new SDH inhibitors using homology modeling; pharmacophore modeling and structure based virtual screening. The three dimensional SDH model showed good stereo-chemical and structural properties. Based on virtual screening results twelve commercially available compounds were purchased and tested in vitro and in vivo. The compounds were found to inhibit mycelial growth of A. solani. Moreover in vitro trials showed that inhibitory effects were enhanced with increase in concentrations. Similarly increased disease control was observed in pre-treated potato tubers. Hence the applied in silico strategy led us to identify novel fungicides. PMID:29204422

Seed isolates of Alternaria and Aspergillus fungi increase germination of Astragalus utahensis

Treesearch

Sean D. Eldredge; Brad Geary; Scott L. Jensen

2016-01-01

Astragalus utahensis (Torr.) Torr. & A. Gray (Fabaceae) (Utah milkvetch) is native lo the arid Great Basin and has desirable attributes that make it a good candidate for restoration in arid, noncompetitive situations. Seed dormancy is a significant barrier to consistent establishment for this species. Species of Alternaria and Aspergillus fungi have...

Genetic transformation of sweet orange with the coat protein gene of Citrus psorosis virus and evaluation of resistance against the virus.

PubMed

Zanek, María Cecilia; Reyes, Carina Andrea; Cervera, Magdalena; Peña, Eduardo José; Velázquez, Karelia; Costa, Norma; Plata, Maria Inés; Grau, Oscar; Peña, Leandro; García, María Laura

2008-01-01

Citrus psorosis is a serious viral disease affecting citrus trees in many countries. Its causal agent is Citrus psorosis virus (CPsV), the type member of genus Ophiovirus. CPsV infects most important citrus varieties, including oranges, mandarins and grapefruits, as well as hybrids and citrus relatives used as rootstocks. Certification programs have not been sufficient to control the disease and no sources of natural resistance have been found. Pathogen-derived resistance (PDR) can provide an efficient alternative to control viral diseases in their hosts. For this purpose, we have produced 21 independent lines of sweet orange expressing the coat protein gene of CPsV and five of them were challenged with the homologous CPV 4 isolate. Two different viral loads were evaluated to challenge the transgenic plants, but so far, no resistance or tolerance has been found in any line after 1 year of observations. In contrast, after inoculation all lines showed characteristic symptoms of psorosis in the greenhouse. The transgenic lines expressed low and variable amounts of the cp gene and no correlation was found between copy number and transgene expression. One line contained three copies of the cp gene, expressed low amounts of the mRNA and no coat protein. The ORF was cytosine methylated suggesting a PTGS mechanism, although the transformant failed to protect against the viral load used. Possible causes for the failed protection against the CPsV are discussed.

7 CFR 301.75-5 - Commercial citrus-producing areas.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 5 2011-01-01 2011-01-01 false Commercial citrus-producing areas. 301.75-5 Section... and Regulations § 301.75-5 Commercial citrus-producing areas. (a) The following are designated as commercial citrus-producing areas: American Samoa Arizona California Florida Guam Hawaii Louisiana Northern...

7 CFR 301.75-5 - Commercial citrus-producing areas.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 5 2014-01-01 2014-01-01 false Commercial citrus-producing areas. 301.75-5 Section... and Regulations § 301.75-5 Commercial citrus-producing areas. (a) The following are designated as commercial citrus-producing areas: American Samoa Arizona California Florida Guam Hawaii Louisiana Northern...

Expression and functional analysis of citrus carotene hydroxylases: unravelling the xanthophyll biosynthesis in citrus fruits.

PubMed

Ma, Gang; Zhang, Lancui; Yungyuen, Witchulada; Tsukamoto, Issei; Iijima, Natsumi; Oikawa, Michiru; Yamawaki, Kazuki; Yahata, Masaki; Kato, Masaya

2016-06-29

Xanthophylls are oxygenated carotenoids and fulfill critical roles in plant growth and development. In plants, two different types of carotene hydroxylases, non-heme di-iron and heme-containing cytochrome P450, were reported to be involved in the biosynthesis of xanthophyll. Citrus fruits accumulate a high amount of xanthophylls, especially β,β-xanthophylls. To date, however, the roles of carotene hydroxylases in regulating xanthophyll content and composition have not been elucidated. In the present study, the roles of four carotene hydroxylase genes (CitHYb, CitCYP97A, CitCYP97B, and CitCYP97C) in the biosynthesis of xanthophyll in citrus fruits were investigated. Phylogenetic analysis showed that the four citrus carotene hydroxylases presented in four distinct clusters which have been identified in higher plants. CitHYb was a non-heme di-iron carotene hydroxylase, while CitCYP97A, CitCYP97B, and CitCYP97C were heme-containing cytochrome P450-type carotene hydroxylases. Gene expression results showed that the expression of CitHYb increased in the flavedo and juice sacs during the ripening process, which was well consistent with the accumulation of β,β-xanthophyll in citrus fruits. The expression of CitCYP97A and CitCYP97C increased with a peak in November, which might lead to an increase of lutein in the juice sacs during the ripening process. The expression level of CitCYP97B was much lower than that of CitHYb, CitCYP97A, and CitCYP97C in the juice sacs during the ripening process. Functional analysis showed that the CitHYb was able to catalyze the hydroxylation of the β-rings of β-carotene and α-carotene in Escherichia coli BL21 (DE3) cells. Meanwhile, when CitHYb was co-expressed with CitCYP97C, α-carotene was hydroxylated on the β-ring and ε-ring sequentially to produce lutein. CitHYb was a key gene for β,β-xanthophyll biosynthesis in citrus fruits. CitCYP97C functioned as an ε-ring hydroxylase to produce lutein using zeinoxanthin as a substrate

Evaluating citrus germplasm for huanglongbing (HLB) resistance: USDA-ARS Inoculation Program

USDA-ARS?s Scientific Manuscript database

The Asian citrus psyllid (ACP), Diaphorina citri, is an important pest because it vectors bacteria responsible for a serious disease of citrus known as huanglongbing (citrus greening disease). USDA-ARS researchers recently established a program for screening citrus germplasm for resistance to the di...

Mechanical Damage Detection of Indonesia Local Citrus Based on Fluorescence Imaging

NASA Astrophysics Data System (ADS)

Siregar, T. H.; Ahmad, U.; Sutrisno; Maddu, A.

2018-05-01

Citrus experienced physical damage in peel will produce essential oils that contain polymethoxylated flavone. Polymethoxylated flavone is fluorescence substance; thus can be detected by fluorescence imaging. This study aims to study the fluorescence spectra characteristic and to determine the damage region in citrus peel based on fluorescence image. Pulung citrus from Batu district, East Java, as a famous citrus production area in Indonesia, was used in the experiment. It was observed that the image processing could detect the mechanical damage region. Fluorescence imaging can be used to classify the citrus into two categories, sound and defect citruses.

Quantitation of flavonoid constituents in citrus fruits.

PubMed

Kawaii, S; Tomono, Y; Katase, E; Ogawa, K; Yano, M

1999-09-01

Twenty-four flavonoids have been determined in 66 Citrus species and near-citrus relatives, grown in the same field and year, by means of reversed phase high-performance liquid chromatography analysis. Statistical methods have been applied to find relations among the species. The F ratios of 21 flavonoids obtained by applying ANOVA analysis are significant, indicating that a classification of the species using these variables is reasonable to pursue. Principal component analysis revealed that the distributions of Citrus species belonging to different classes were largely in accordance with Tanaka's classification system.

Enhancement of β-Carotene Synthesis by Citrus Products

PubMed Central

Ciegler, Alex; Nelson, George E. N.; Hall, Harlow H.

1963-01-01

β-Ionone, a stimulatory compound in the microbiological production of β-carotene by mated cultures of Blakeslea trispora, could be replaced with low-cost agricultural by-products (citrus oils, citrus pulp, or citrus molasses) with as good or better carotene yields. Peak yields (81 to 129 mg of carotene per g of dry solids) were achieved in 5 days. The various citrus products tested did not change the pigments produced; all trans-β-carotene remained the pre-dominant pigment. The acid-hydrolyzed soybean meal and corn used in previous production media could be replaced with unhydrolyzed cottonseed embryo meal and corn in a medium that also contained a natural lipid, deodorized kerosene, nonionic detergent, and a precursor. PMID:14021337

Citrus peel extract incorporated ice cubes to protect the quality of common pandora: Fish storage in ice with citrus.

PubMed

Yerlikaya, Pinar; Ucak, Ilknur; Gumus, Bahar; Gokoglu, Nalan

2015-12-01

The objective of this study was to investigate the effects of ice with albedo and flavedo fragments of Citrus (Grapefruit (Citrus paradisi) and Bitter orange (Citrus aurantium L.)) extracts on the quality of common pandora (Pagellus erythrinus). Concentrated citrus extracts were diluted with distilled water (1/100 w/v) before making of ice. The ice cubes were spread on each layer of fishes and stored at 0 °C for 15 days. The pH value showed a regular increase in all samples. TVB-N levels of bitter orange treatment groups were recorded lower than the other groups reaching to 25.11 ± 0.02 mg/100 g at the end of the storage. The TMA-N values of bitter orange treatment groups were lower than that of control and grapefruit treatment groups. In terms of TBARS value, alteration was observed in the control samples and this value significantly (p < 0.01) increased from 0.101 ± 0.011 mg MA/kg to 0.495 ± 0.083 mg MA/kg, while remained lower in the citrus extracts treatment groups at the end of storage since their antioxidant capacity. The oxidation was suppressed in citrus extracts treatment groups, especially in bitter orange flavedo treatment. The results showed the bitter orange albedo and bitter orange flavedo extracts in combination with ice storage have more effectiveness in controlling the biochemical indices in common pandora.

Changes in the leaf proteome profile of Withania somnifera (L.) Dunal in response to Alternaria alternata infection

PubMed Central

Singh, Varinder; Singh, Baldev; Joshi, Robin; Jaju, Puneet

2017-01-01

Withania somnifera is a high value medicinal plant which is used against large number of ailments. The medicinal properties of the plant attributes to a wide array of important secondary metabolites. The plant is predominantly infected with leaf spot pathogen Alternaria alternata, which leads to substantial biodeterioration of pharmaceutically important metabolites. To develop an effective strategy to combat this disease, proteomics based approach could be useful. Hence, in the present study, three different protein extraction methods tris-buffer based, phenol based and trichloroacetic acid-acetone (TCA-acetone) based method were comparatively evaluated for two-dimensional electrophoresis (2-DE) analysis of W. somnifera. TCA-acetone method was found to be most effective and was further used to identify differentially expressed proteins in response to fungal infection. Thirty-eight differentially expressed proteins were identified by matrix assisted laser desorption/ionization time of flight-mass spectrometry (MALDI TOF/TOF MS/MS). The known proteins were categorized into eight different groups based on their function and maximum proteins belonged to energy and metabolism, cell structure, stress and defense and RNA/DNA categories. Differential expression of some key proteins were also crosschecked at transcriptomic level by using qRT-PCR and were found to be consistent with the 2-DE data. These outcomes enable us to evaluate modifications that take place at the proteomic level during a compatible host pathogen interaction. The comparative proteome analysis conducted in this paper revealed the involvement of many key proteins in the process of pathogenesis and further investigation of these identified proteins could assist in the discovery of new strategies for the development of pathogen resistance in the plant. PMID:28575108

Immunoblotting Quantification Approach for Identifying Potential Hypoallergenic Citrus Cultivars.

PubMed

Wu, Jinlong; Deng, Wenjun; Lin, Dingbo; Deng, Xiuxin; Ma, Zhaocheng

2018-02-28

The inherent allergens of citrus fruits, such as Cit s 1, Cit s 2, Cit s 3 can cause allergic reactions. A better understanding of the genetic factors (cultivar to cultivar) affecting the allergenic potential of citrus fruits would be beneficial for further identification of hypoallergenic genotypes. In the present study, an immunoblotting quantification approach was adopted to assess the potential allergenicity of 21 citrus cultivars, including nine subgroups (tangerine, satsuma, orange, pummelo, grapefruit, lemon, kumquat, tangor, and tangelo). To prepare highly sensitive and specific rabbit polyclonal antibodies, antigenicity of purified rCit s 1.01, rCit s 2.01, and rCit s 3.01 peptides were enhanced with high epitope density in a single protein molecule. The data integration of three citrus allergen quantifications demonstrated that the four pummelo cultivars (Kao Phuang Pummelo, Wanbai Pummelo, Shatian Pummelo, and Guanxi Pummelo) were potential hypoallergenic, compared with other 8 subgroups. Moreover, the immunological analyses with sera of allergic subjects revealed that Shatian Pummelo and Guanxi Pummelo showed the lowest immunoreactivity in 8 representative citrus cultivars. These potential hypoallergenic genotypes are of great significance to not only allergic consumers but also citrus breeders in the genetic improvement of hypoallergenic citrus as breeding resources.

Bioavailable Citrus sinensis Extract: Polyphenolic Composition and Biological Activity.

PubMed

Pepe, Giacomo; Pagano, Francesco; Adesso, Simona; Sommella, Eduardo; Ostacolo, Carmine; Manfra, Michele; Chieppa, Marcello; Sala, Marina; Russo, Mariateresa; Marzocco, Stefania; Campiglia, Pietro

2017-04-15

Citrus plants contain large amounts of flavonoids with beneficial effects on human health. In the present study, the antioxidant and anti-inflammatory potential of bioavailable polyphenols from Citrus sinensis was evaluated in vitro and ex vivo, using the murine macrophages cell line J774A.1 and primary peritoneal macrophages. Following simulated gastro-intestinal digestion, the in vitro bioavailability of Citrus sinensis polyphenolic extract was assessed using the human cell line Caco-2 grown as monolayers on a transwell membrane. Data demonstrated a relative permeation of its compounds (8.3%). Thus, the antioxidant and anti-inflammatory effect of polyphenolic Citrus sinensis fraction (Cs) was compared to the bioavailable one (CsB). Results revealed that Citrus extract were able to reduce macrophages pro-inflammatory mediators, including nitric oxide, iNOS, COX-2 and different cytokines. Moreover, the effect of Citrus sinensis polyphenols was associated with antioxidant effects, such as a reduction of reactive oxygen species (ROS) and heme-oxygenase-1 (HO-1) increased expression. Our results provide evidence that the bioavailable polyphenolic constituents of the Citrus sinensis extract accumulate prevalently at intestinal level and could reach systemic circulation exerting their effect. The bioavailable fraction showed a higher anti-inflammatory and antioxidant potential compared to the initial extract, thus highlighting its potential nutraceutical value.

Infection of an Insect Vector with a Bacterial Plant Pathogen Increases Its Propensity for Dispersal

PubMed Central

Coy, Monique R.; Stelinski, Lukasz L.; Pelz-Stelinski, Kirsten S.

2015-01-01

The spread of vector-transmitted pathogens relies on complex interactions between host, vector and pathogen. In sessile plant pathosystems, the spread of a pathogen highly depends on the movement and mobility of the vector. However, questions remain as to whether and how pathogen-induced vector manipulations may affect the spread of a plant pathogen. Here we report for the first time that infection with a bacterial plant pathogen increases the probability of vector dispersal, and that such movement of vectors is likely manipulated by a bacterial plant pathogen. We investigated how Candidatus Liberibacter asiaticus (CLas) affects dispersal behavior, flight capacity, and the sexual attraction of its vector, the Asian citrus psyllid (Diaphorina citri Kuwayama). CLas is the putative causal agent of huanglongbing (HLB), which is a disease that threatens the viability of commercial citrus production worldwide. When D. citri developed on CLas-infected plants, short distance dispersal of male D. citri was greater compared to counterparts reared on uninfected plants. Flight by CLas-infected D. citri was initiated earlier and long flight events were more common than by uninfected psyllids, as measured by a flight mill apparatus. Additionally, CLas titers were higher among psyllids that performed long flights than psyllid that performed short flights. Finally, attractiveness of female D. citri that developed on infected plants to male conspecifics increased proportionally with increasing CLas bacterial titers measured within female psyllids. Our study indicates that the phytopathogen, CLas, may manipulate movement and mate selection behavior of their vectors, which is a possible evolved mechanism to promote their own spread. These results have global implications for both current HLB models of disease spread and control strategies. PMID:26083763

Occurrence of Cladosporium spp. and Alternaria spp. spores in Western, Northern and Central-Eastern Poland in 2004-2006 and relation to some meteorological factors

NASA Astrophysics Data System (ADS)

Grinn-Gofroń, Agnieszka; Rapiejko, Piotr

2009-08-01

The concentration of airborne spores of Cladosporium spp. and Alternaria spp. has been investigated at three monitoring stations situated along the west-north and central-east transect in Poland (Szczecin, Olsztyn, Warszawa,) i.e. from a height of 100 m to 149 m above sea level. The aerobiological monitoring of fungal spores was performed by means of three Lanzoni volumetric spore traps. Cladosporium spp. spores were dominant at all the stations. The highest Cladosporium spp. and Alternaria spp. numbers of spores were observed at all the cities in July and August. Statistically significant correlations have been found between the Cladosporium spp. and Alternaria spp. concentration in the air and the mean air temperature, amount of precipitation, air pressure and relative air humidity. The spore count of Cladosporium spp. and Alternaria spp. is determined by the diversity of local flora and weather conditions, especially by the air temperature. The identification of factors, which influence and shape spore concentrations, may significantly improve the current methods of allergy prevention.

The information extraction of Gannan citrus orchard based on the GF-1 remote sensing image

NASA Astrophysics Data System (ADS)

Wang, S.; Chen, Y. L.

2017-02-01

The production of Gannan oranges is the largest in China, which occupied an important part in the world. The extraction of citrus orchard quickly and effectively has important significance for fruit pathogen defense, fruit production and industrial planning. The traditional spectra extraction method of citrus orchard based on pixel has a lower classification accuracy, difficult to avoid the “pepper phenomenon”. In the influence of noise, the phenomenon that different spectrums of objects have the same spectrum is graveness. Taking Xunwu County citrus fruit planting area of Ganzhou as the research object, aiming at the disadvantage of the lower accuracy of the traditional method based on image element classification method, a decision tree classification method based on object-oriented rule set is proposed. Firstly, multi-scale segmentation is performed on the GF-1 remote sensing image data of the study area. Subsequently the sample objects are selected for statistical analysis of spectral features and geometric features. Finally, combined with the concept of decision tree classification, a variety of empirical values of single band threshold, NDVI, band combination and object geometry characteristics are used hierarchically to execute the information extraction of the research area, and multi-scale segmentation and hierarchical decision tree classification is implemented. The classification results are verified with the confusion matrix, and the overall Kappa index is 87.91%.

Dietary citrus pulp reduces lipid oxidation in lamb meat.

PubMed

Inserra, L; Priolo, A; Biondi, L; Lanza, M; Bognanno, M; Gravador, R; Luciano, G

2014-04-01

This study investigated the effect of replacing cereal concentrates with high levels of dried citrus pulp in the diet on lamb meat oxidative stability. Over 56 days, lambs were fed a barley-based concentrate (Control) or concentrates in which 24% and 35% dried citrus pulp were included to partially replace barley (Citrus 24% and Citrus 35%, respectively). Meat was aged under vacuum for 4 days and subsequently stored aerobically at 4 °C. The Control diet increased the redness, yellowness and saturation of meat after blooming (P<0.01). Regardless of the level of supplementation, dietary dried citrus pulp strongly reduced meat lipid oxidation over 6 days of aerobic storage (P<0.001), while colour parameters did not change noticeably over storage and their variation rate was not affected by the diet. In conclusion, replacing cereals with dried citrus pulp in concentrate-based diets might represent a feasible strategy to naturally improve meat oxidative stability and to promote the exploitation of this by-product. Copyright © 2013 Elsevier Ltd. All rights reserved.

Expression patterns of flowering genes in leaves of 'Pineapple' sweet orange [Citrus sinensis (L.) Osbeck] and pummelo (Citrus grandis Osbeck).

PubMed

Pajon, Melanie; Febres, Vicente J; Moore, Gloria A

2017-08-30

In citrus the transition from juvenility to mature phase is marked by the capability of a tree to flower and fruit consistently. The long period of juvenility in citrus severely impedes the use of genetic based strategies to improve fruit quality, disease resistance, and responses to abiotic environmental factors. One of the genes whose expression signals flower development in many plant species is FLOWERING LOCUS T (FT). In this study, gene expression levels of flowering genes CiFT1, CiFT2 and CiFT3 were determined using reverse-transcription quantitative real-time PCR in citrus trees over a 1 year period in Florida. Distinct genotypes of citrus trees of different ages were used. In mature trees of pummelo (Citrus grandis Osbeck) and 'Pineapple' sweet orange (Citrus sinensis (L.) Osbeck) the expression of all three CiFT genes was coordinated and significantly higher in April, after flowering was over, regardless of whether they were in the greenhouse or in the field. Interestingly, immature 'Pineapple' seedlings showed significantly high levels of CiFT3 expression in April and June, while CiFT1 and CiFT2 were highest in June, and hence their expression induction was not simultaneous as in mature plants. In mature citrus trees the induction of CiFTs expression in leaves occurs at the end of spring and after flowering has taken place suggesting it is not associated with dormancy interruption and further flower bud development but is probably involved with shoot apex differentiation and flower bud determination. CiFTs were also seasonally induced in immature seedlings, indicating that additional factors must be suppressing flowering induction and their expression has other functions.

Biocontrol Potential of Streptomyces hydrogenans Strain DH16 toward Alternaria brassicicola to Control Damping Off and Black Leaf Spot of Raphanus sativus

PubMed Central

Manhas, Rajesh K.; Kaur, Talwinder

2016-01-01

Biocontrol agents and their bioactive metabolites provide one of the best alternatives to decrease the use of chemical pesticides. In light of this, the present investigation reports the biocontrol potential of Streptomyces hydrogenans DH16 and its metabolites towards Alternaria brassicicola, causal agent of black leaf spot and damping off of seedlings of crucifers. In vitro antibiosis of strain against pathogen revealed complete suppression of mycelial growth of pathogen, grown in potato dextrose broth supplemented with culture supernatant (20% v/v) of S. hydrogenans DH16. Microscopic examination of the fungal growth showed severe morphological abnormalities in the mycelium caused by antifungal metabolites. In vivo studies showed the efficacy of streptomycete cells and culture supernatant as seed dressings to control damping off of Raphanus sativus seedlings. Treatment of pathogen infested seeds with culture supernatant (10%) and streptomycete cells significantly improved seed germination (75–80%) and vigor index (1167–1538). Furthermore, potential of cells and culture supernatant as foliar treatment to control black leaf spot was also evaluated. Clearly visible symptoms of disease were observed in the control plants with 66.81% disease incidence and retarded growth of root system. However, disease incidence reduced to 6.78 and 1.47% in plants treated with antagonist and its metabolites, respectively. Additionally, treatment of seeds and plants with streptomycete stimulated various growth traits of plants over uninoculated control plants in the absence of pathogen challenge. These results indicate that S. hydrogenans and its culture metabolites can be developed as biofungicides as seed dressings to control seed borne pathogens, and as sprays to control black leaf spot of crucifers. PMID:28018402

Resistance and tolerance to Huanglongbing in citrus

USDA-ARS?s Scientific Manuscript database

The Stover-led research team focuses on developing improved citrus scion varieties, through both conventional breeding and transgenics, with special attention to confronting the threat of citrus huanglongbing (HLB). Collaborations are underway with more than 20 research groups around the world to ha...

Regulation of cuticle formation during fruit development and ripening in 'Newhall' navel orange (Citrus sinensis Osbeck) revealed by transcriptomic and metabolomic profiling.

PubMed

Wang, Jinqiu; Sun, Li; Xie, Li; He, Yizhong; Luo, Tao; Sheng, Ling; Luo, Yi; Zeng, Yunliu; Xu, Juan; Deng, Xiuxin; Cheng, Yunjiang

2016-02-01

Fruit cuticle, which is composed of cutin and wax and biosynthesized during fruit development, plays important roles in the prevention of water loss and the resistance to pathogen infection during fruit development and postharvest storage. However, the key factors and mechanisms regarding the cuticle biosynthesis in citrus fruits are still unclear. Here, fruit cuticle of 'Newhall' navel orange (Citrus sinensis Osbeck) was studied from the stage of fruit expansion to postharvest storage from the perspectives of morphology, transcription and metabolism. The results demonstrated that cutin accumulation is synchronous with fruit expansion, while wax synthesis is synchronous with fruit maturation. Metabolic profile of fruits peel revealed that transition of metabolism of fruit peel occurred from 120 to 150 DAF and ABA was predicted to regulate citrus wax synthesis during the development of Newhall fruits. RNA-seq analysis of the peel from the above two stages manifested that the genes involved in photosynthesis were repressed, while the genes involved in the biosynthesis of wax, cutin and lignin were significantly induced at later stages. Further real-time PCR predicted that MYB transcription factor GL1-like regulates citrus fruits wax synthesis. These results are valuable for improving the fruit quality during development and storage. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Alt a 1 allergen homologs from Alternaria and related taxa: analysis of phylogenetic content and secondary structure.

PubMed

Hong, Soon Gyu; Cramer, Robert A; Lawrence, Christopher B; Pryor, Barry M

2005-02-01

A gene for the Alternaria major allergen, Alt a 1, was amplified from 52 species of Alternaria and related genera, and sequence information was used for phylogenetic study. Alt a 1 gene sequences evolved 3.8 times faster and contained 3.5 times more parsimony-informative sites than glyceraldehyde-3-phosphate dehydrogenase (gpd) sequences. Analyses of Alt a 1 gene and gpd exon sequences strongly supported grouping of Alternaria spp. and related taxa into several species-groups described in previous studies, especially the infectoria, alternata, porri, brassicicola, and radicina species-groups and the Embellisia group. The sonchi species-group was newly suggested in this study. Monophyly of the Nimbya group was moderately supported, and monophyly of the Ulocladium group was weakly supported. Relationships among species-groups and among closely related species of the same species-group were not fully resolved. However, higher resolution could be obtained using Alt a 1 sequences or a combined dataset than using gpd sequences alone. Despite high levels of variation in amino acid sequences, results of in silico prediction of protein secondary structure for Alt a 1 demonstrated a high degree of structural similarity for most of the species suggesting a conservation of function.

Antifungal activity of altenusin isolated from the endophytic fungus Alternaria sp. against the pathogenic fungus Paracoccidioides brasiliensis.

PubMed

Johann, Susana; Rosa, Luiz H; Rosa, Carlos A; Perez, Pilar; Cisalpino, Patrícia S; Zani, Carlos L; Cota, Betania B

2012-01-01

Altenusin is a biphenyl derivative isolated from different species of fungi, which presents several biological activities. We report the antifungal activity of the altenusin isolated from the endophytic fungus Alternaria sp., against clinical isolates of Paracoccidioides brasiliensis, and its action on cell walls of P. brasiliensis and the nonpathogenic yeast Schizosaccharomyces pombe. In vitro antifungal activity of altenusin was evaluated using the broth microdilution method against 11 strains of P. brasiliensis and one strain of S. pombe. The effects of the altenusin on the cell wall were estimated using the sorbitol protection assay. The altenusin presented strong activity against P. brasiliensis with MIC values ranging between 1.9 and 31.2 μg/ml, and 62.5 μg/ml for S. pombe. Our results demonstrated that the MIC values for altenusin were increased for P. brasiliensis Pb18 and for S. pombe when the medium was supplemented with sorbitol. Additionally, S. pombe cells treated with altenusin were more rounded in shape than untreated cells. Altenusin showed activity against clinical strains of P. brasiliensis at the concentration tested, and this compound probably affects fungal cell walls. These findings suggest that altenusin could act through the inhibition of cell wall synthesis or assembly in P. brasiliensis and S. pombe, and could be considered as a lead compound for the design of new antifungals. Copyright © 2011 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

Transcriptome analysis of sweet orange trees infected with 'Candidatus Liberibacter asiaticus' and two strains of Citrus Tristeza Virus.

PubMed

Fu, Shimin; Shao, Jonathan; Zhou, Changyong; Hartung, John S

2016-05-11

Huanglongbing (HLB) and tristeza, are diseases of citrus caused by a member of the α-proteobacteria, 'Candidatus Liberibacter asiaticus' (CaLas), and Citrus tristeza virus (CTV) respectively. HLB is a devastating disease, but CTV strains vary from very severe to very mild. Both CaLas and CTV are phloem-restricted. The CaLas-B232 strain and CTV-B6 cause a wide range of severe and similar symptoms. The mild strain CTV-B2 doesn't induce significant symptoms or damage to plants. Transcriptome profiles obtained through RNA-seq revealed 611, 404 and 285 differentially expressed transcripts (DETs) after infection with CaLas-B232, CTV-B6 and CTV-B2. These DETs were components of a wide range of pathways involved in circadian rhythm, cell wall modification and cell organization, as well as transcription factors, transport, hormone response and secondary metabolism, signaling and stress response. The number of transcripts that responded to both CTV-B6 and CaLas-B232 was much larger than the number of transcripts that responded to both strains of CTV or to both CTV-B2 and CaLas-B232. A total of 38 genes were assayed by RT-qPCR and the correlation coefficients between Gfold and RT-qPCR were 0.82, 0.69, 0.81 for sweet orange plants infected with CTV-B2, CTV-B6 and CaLas-B232, respectively. The number and composition of DETs reflected the complexity of symptoms caused by the pathogens in established infections, although the leaf tissues sampled were asymptomatic. There were greater similarities between the sweet orange in response to CTV-B6 and CaLas-B232 than between the two CTV strains, reflecting the similar physiological changes caused by both CTV-B6 and CaLas-B232. The circadian rhythm system of plants was perturbed by all three pathogens, especially by CTV-B6, and the ion balance was also disrupted by all three pathogens, especially by CaLas-B232. Defense responses related to cell wall modification, transcriptional regulation, hormones, secondary metabolites, kinases and

Anti-inflammatory effects of Citrus sinensis L., Citrus paradisi L. and their combinations.

PubMed

Khan, Rafeeq Alam; Mallick, Neelam; Feroz, Zeeshan

2016-05-01

Citrus bioflavonoids embrace a wide group of phenolic compounds effecting the production and scavenging of reactive oxygen species and the processes relating free radical-mediated injury. Keeping in view of the antioxidant and anti-inflammatory properties of Citrus sinensis and Citrus paradisi, present study was undertaken to explore the effects of C. sinensis (orange juice) and C. paradisi (grapefruit juice) at three different doses alone and their two combinations with the objective to examine the effects of these compounds in an experimental model of rat colitis induced by trinitrobenzenesulphonic acid (TNBS). Hence biochemical parameters e.g. myeloperoxidase, alkaline phosphatase, C-reactive protein (CRP) and glutathione were assessed. Data entry and analysis was accomplished by Statistical Package for the Social Sciences version 17 and was presented as mean ± S.E.M with 95% confidence interval. Present result shows that these juices, mainly C. paradisi, may be efficacious for the management of inflammatory bowel disease. In acute colitis model, C. paradise encouraged a decrease in the extension of the lesion escorted by a decrease in the occurrence of diarrhea and reinstatement of the glutathione content. Related effects were produced by the administration of C. sinensis, which also prevented the myeloperoxidase and alkaline phosphatase actions in acute intestinal inflammatory process. The effect of the citrus juices on the inflammatory process may be associated to their antioxidant and anti-inflammatory properties, as revealed in present investigation. The favorable effects exerted were demonstrated both by histological and biochemical changes and were related with a progress in the colonic oxidative status.

Asian citrus psyllid RNAi pathway - RNAi evidence

USDA-ARS?s Scientific Manuscript database

In silico analyses of the draft genome of Diaphorina citri, the Asian citrus psyllid, for genes within the Ribonucleic acid interference(RNAi), pathway was successful. The psyllid is the vector of the plant-infecting bacterium, Candidatus Liberibacter asiaticus (CLas), which is linked to citrus gree...

76 FR 23449 - Citrus Canker, Citrus Greening, and Asian Citrus Psyllid; Interstate Movement of Regulated...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-27

... conditions, the articles are treated with methyl bromide and shipped in a container that has been sealed with... of any regulated article, including citrus nursery stock, provided that: The article is treated with methyl bromide in accordance with 7 CFR part 305. That part contains our phytosanitary treatment...

Somatic Embryogenesis: Still a Relevant Technique in Citrus Improvement.

PubMed

Omar, Ahmad A; Dutt, Manjul; Gmitter, Frederick G; Grosser, Jude W

2016-01-01

The genus Citrus contains numerous fresh and processed fruit cultivars that are economically important worldwide. New cultivars are needed to battle industry threatening diseases and to create new marketing opportunities. Citrus improvement by conventional methods alone has many limitations that can be overcome by applications of emerging biotechnologies, generally requiring cell to plant regeneration. Many citrus genotypes are amenable to somatic embryogenesis, which became a key regeneration pathway in many experimental approaches to cultivar improvement. This chapter provides a brief history of plant somatic embryogenesis with focus on citrus, followed by a discussion of proven applications in biotechnology-facilitated citrus improvement techniques, such as somatic hybridization, somatic cybridization, genetic transformation, and the exploitation of somaclonal variation. Finally, two important new protocols that feature plant regeneration via somatic embryogenesis are provided: protoplast transformation and Agrobacterium-mediated transformation of embryogenic cell suspension cultures.

Genome-wide genetic variation and comparison of fruit-associated traits between kumquat (Citrus japonica) and Clementine mandarin (Citrus clementina).

PubMed

Liu, Tian-Jia; Li, Yong-Ping; Zhou, Jing-Jing; Hu, Chun-Gen; Zhang, Jin-Zhi

2018-03-01

The comprehensive genetic variation of two citrus species were analyzed at genome and transcriptome level. A total of 1090 differentially expressed genes were found during fruit development by RNA-sequencing. Fruit size (fruit equatorial diameter) and weight (fresh weight) are the two most important components determining yield and consumer acceptability for many horticultural crops. However, little is known about the genetic control of these traits. Here, we performed whole-genome resequencing to reveal the comprehensive genetic variation of the fruit development between kumquat (Citrus japonica) and Clementine mandarin (Citrus clementina). In total, 5,865,235 single-nucleotide polymorphisms (SNPs) and 414,447 insertions/deletions (InDels) were identified in the two citrus species. Based on integrative analysis of genome and transcriptome of fruit, 640,801 SNPs and 20,733 InDels were identified. The features, genomic distribution, functional effect, and other characteristics of these genetic variations were explored. RNA-sequencing identified 1090 differentially expressed genes (DEGs) during fruit development of kumquat and Clementine mandarin. Gene Ontology revealed that these genes were involved in various molecular functional and biological processes. In addition, the genetic variation of 939 DEGs and 74 multiple fruit development pathway genes from previous reports were also identified. A global survey identified 24,237 specific alternative splicing events in the two citrus species and showed that intron retention is the most prevalent pattern of alternative splicing. These genome variation data provide a foundation for further exploration of citrus diversity and gene-phenotype relationships and for future research on molecular breeding to improve kumquat, Clementine mandarin and related species.

Development of infectious cDNA clones of citrus yellow vein clearing virus using a novel and rapid strategy.

PubMed

Cui, Tian Tian; Bin, Yu; Yan, Jian Hong; Mei, Peng Ying; Li, Zhong An; Zhou, Chang Yong; Song, Zhen

2018-05-04

Yellow vein clearing disease (YVCD) causes significant economic losses in lemon and other species of citrus. Usually, citrus yellow vein clearing virus (CYVCV) is considered to be the causal agent of YVCD. However, mixed infection of CYVCV and Indian citrus ringspot virus (ICRSV) or other pathogens is often detected in citrus plants with YVCD. In this study, we re-examined the causal agent of YVCD to fulfill Koch's postulates. First, the full-length genome of CYVCV isolate AY (CYVCV-AY) was amplified by long-distance RT-PCR from a Eureka lemon [Citrus limon (L) Brum. f.] tree with typical YVCD symptoms. The genomic cDNAs were then cloned into a ternary Yeast-Escherichia coli-Agrobacterium tumefaciens shuttle vector, pCY, using transformation-associated recombination (TAR) strategy, and 15 full-length cDNA clones of CYVCV-AY were obtained. Subsequently, four of these clones were selected randomly and inoculated on Jincheng [C. sinensis (L) Osbeck] seedlings through Agrobacterium-mediated vacuum-infiltration, and it was found that 80 to 100% of inoculated plants were infected with CYVCV by RT-PCR at 20 to 40 days post inoculation (dpi) and by direct tissue blot immunoassay at 60 dpi. The progeny of CYVCV-AY from cDNA clones caused typical symptoms of YVCD such as yellow vein clearing, leaf distortion, and chlorosis, which were the same as that elicited by wild-type virus. Finally, the regeneration of CYVCV-AY genome was confirmed by long-distance RT-PCR in lemon trees inoculated with the infectious cDNA clone. These results proved that CYVCV was the primary causal agent of YVCD. This is the first report on the development of infectious cDNA clones of CYVCV, which lays the foundation for further studies on viral gene functions and virus-host interactions.

Metalized polyethylene mulch to repel Asian citrus psyllid, slow spread of huanglongbing and improve growth of new citrus plantings.

PubMed

Croxton, Scott D; Stansly, Philip A

2014-02-01

Greening or huanglongbing (HLB) is a debilitating disease of citrus caused by Candidatus Liberibactor asiaticus and transmitted by the Asian citrus psyllid (ACP), Diaphorina citri. HLB now occurs worldwide in all major citrus growing regions except the Mediterranean and Australia. Management relies principally on insecticidal control of the ACP vector, but is insufficient, even for young trees which are most susceptible to the disease. We tested the ability of metalized polyethylene mulch to repel adult ACP as well as effects on incidence of HLB and early tree growth. Metalized mulch significantly reduced ACP populations and HLB incidence compared to whiteface mulch or bare ground. In addition, metalized mulch, together with the associated drip irrigation and fertigation system, increased soil moisture, reduced weed pressure, and increased tree growth rate. Metalized mulch slows spread of ACP and therefore HLB pressure on young citrus trees. Metalized mulch can thereby augment current control measures for young trees based primarily on systemic insecticides. Additional costs could be compensated for by increased tree growth rate which would shorten time to crop profitability. These advantages make a compelling case for large-scale trials using metalized mulch in young citrus plantings threatened by HLB. © 2013 Society of Chemical Industry.

Citrus fruit recognition using color image analysis

NASA Astrophysics Data System (ADS)

Xu, Huirong; Ying, Yibin

2004-10-01

An algorithm for the automatic recognition of citrus fruit on the tree was developed. Citrus fruits have different color with leaves and branches portions. Fifty-three color images with natural citrus-grove scenes were digitized and analyzed for red, green, and blue (RGB) color content. The color characteristics of target surfaces (fruits, leaves, or branches) were extracted using the range of interest (ROI) tool. Several types of contrast color indices were designed and tested. In this study, the fruit image was enhanced using the (R-B) contrast color index because results show that the fruit have the highest color difference among the objects in the image. A dynamic threshold function was derived from this color model and used to distinguish citrus fruit from background. The results show that the algorithm worked well under frontlighting or backlighting condition. However, there are misclassifications when the fruit or the background is under a brighter sunlight.

Transmission of Citrus leprosis virus C by Brevipalpus phoenicis (Geijskes) to Alternative Host Plants Found in Citrus Orchards

USDA-ARS?s Scientific Manuscript database

The equivalent of US$ 75 million is spent each year in Brazil to control Brevipalpus phoenicis, a mite vector of Citrus leprosis virus C (CiLV-C). In this study we investigated the possibility that hedgerows, windbreaks, and weeds normally found in citrus orchards could host CiLV-C. Mites reared on ...

Comparative analysis of juice volatiles in selected mandarins, mandarin relatives and other citrus genotypes.

PubMed

Yu, Yuan; Bai, Jinhe; Chen, Chunxian; Plotto, Anne; Baldwin, Elizabeth A; Gmitter, Frederick G

2018-02-01

Citrus fruit flavor is an important attribute prioritized in variety improvement. The present study compared juice volatiles compositions from 13 selected citrus genotypes, including six mandarins (Citrus reticulata), three sour oranges (Citrus aurantium), one blood orange (Citrus sinensis), one lime (Citrus limonia), one Clementine (Citrus clementina) and one satsuma (Citrus unshiu). Large differences were observed with respect to volatile compositions among the citrus genotypes. 'Goutou' sour orange contained the greatest number of volatile compounds and the largest volatile production level. 'Ponkan' mandarin had the smallest number of volatiles and 'Owari' satsuma yielded the lowest volatile production level. 'Goutou' sour orange and 'Moro' blood orange were clearly distinguished from other citrus genotypes based on the analysis of volatile compositions, even though they were assigned into one single group with two other sour oranges by the molecular marker profiles. The clustering analysis based on the aroma volatile compositions was able to differentiate mandarin varieties and natural sub-groups, and was also supported by the molecular marker study. The gas chromatography-mass spectrometry analysis of citrus juice aroma volatiles can be used as a tool to distinguish citrus genotypes and assist in the assessment of future citrus breeding programs. The aroma volatile profiles of the different citrus genotypes and inter-relationships detected among volatile compounds and among citrus genotypes will provide fundamental information on the development of marker-assisted selection in citrus breeding. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Degradation of the Alternaria mycotoxins alternariol, alternariol monomethyl ether, and altenuene upon bread baking.

PubMed

Siegel, David; Feist, Michael; Proske, Matthias; Koch, Matthias; Nehls, Irene

2010-09-08

The stability of the Alternaria mycotoxins alternariol, alternariol monomethyl ether, and altenuene upon bread baking was investigated by model experiments using a spiked wholemeal wheat flour matrix. For alternariol and alternariol monomethyl ether, but not for altenuene, degradation products, formed through a sequence of hydrolysis and decarboxylation, could be identified in pilot studies. The simultaneous quantification of alternariol, alternariol monomethyl ether, altenuene, and the degradation products was achieved by a newly developed high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) multimethod. The obtained quantitative data indicate that the Alternaria mycotoxins are barely degraded during wet baking, while significant degradation occurs upon dry baking, with the stability decreasing in the order alternariol monomethyl ether>alternariol>altenuene. The novel degradation products could be detected after the wet baking of flour spiked with alternariol and in a sample survey of 24 commercial cereal based baking products.

Spectral difference analysis and airborne imaging classification for citrus greening infected trees

USDA-ARS?s Scientific Manuscript database

Citrus greening, also called Huanglongbing (HLB), became a devastating disease spread through citrus groves in Florida, since it was first found in 2005. Multispectral (MS) and hyperspectral (HS) airborne images of citrus groves in Florida were acquired to detect citrus greening infected trees in 20...

7 CFR 457.106 - Texas citrus tree crop insurance provisions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 6 2014-01-01 2014-01-01 false Texas citrus tree crop insurance provisions. 457.106... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.106 Texas citrus tree crop insurance provisions. The Texas Citrus Tree Crop Insurance Provisions for the 2011 and...

7 CFR 457.106 - Texas citrus tree crop insurance provisions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 6 2013-01-01 2013-01-01 false Texas citrus tree crop insurance provisions. 457.106... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.106 Texas citrus tree crop insurance provisions. The Texas Citrus Tree Crop Insurance Provisions for the 2011 and...

7 CFR 457.119 - Texas citrus fruit crop insurance provisions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 6 2012-01-01 2012-01-01 false Texas citrus fruit crop insurance provisions. 457.119... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.119 Texas citrus fruit crop insurance provisions. The Texas citrus fruit crop insurance provisions for the 2000 and...

7 CFR 457.106 - Texas citrus tree crop insurance provisions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 6 2012-01-01 2012-01-01 false Texas citrus tree crop insurance provisions. 457.106... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.106 Texas citrus tree crop insurance provisions. The Texas Citrus Tree Crop Insurance Provisions for the 2011 and...

7 CFR 457.119 - Texas citrus fruit crop insurance provisions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 6 2010-01-01 2010-01-01 false Texas citrus fruit crop insurance provisions. 457.119... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.119 Texas citrus fruit crop insurance provisions. The Texas citrus fruit crop insurance provisions for the 2000 and...

7 CFR 457.106 - Texas citrus tree crop insurance provisions.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 6 2011-01-01 2011-01-01 false Texas citrus tree crop insurance provisions. 457.106... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.106 Texas citrus tree crop insurance provisions. The Texas Citrus Tree Crop Insurance Provisions for the 2011 and...

7 CFR 457.119 - Texas citrus fruit crop insurance provisions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 6 2014-01-01 2014-01-01 false Texas citrus fruit crop insurance provisions. 457.119... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.119 Texas citrus fruit crop insurance provisions. The Texas citrus fruit crop insurance provisions for the 2000 and...

7 CFR 457.119 - Texas citrus fruit crop insurance provisions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 6 2013-01-01 2013-01-01 false Texas citrus fruit crop insurance provisions. 457.119... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.119 Texas citrus fruit crop insurance provisions. The Texas citrus fruit crop insurance provisions for the 2000 and...

7 CFR 457.119 - Texas citrus fruit crop insurance provisions.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 6 2011-01-01 2011-01-01 false Texas citrus fruit crop insurance provisions. 457.119... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.119 Texas citrus fruit crop insurance provisions. The Texas citrus fruit crop insurance provisions for the 2000 and...

Long term field evaluation reveals HLB resistance in Citrus relatives

USDA-ARS?s Scientific Manuscript database

Citrus huanglongbing (HLB) is a destructive disease with no known cure. To identify sources of HLB resistance in the subfamily Aurantioideae to which citrus belongs, we conducted a six-year field trial under natural disease challenge conditions in an HLB endemic region. The study included 65 Citrus ...

Activation of Melanin Synthesis in Alternaria infectoria by Antifungal Drugs.

PubMed

Fernandes, Chantal; Prados-Rosales, Rafael; Silva, Branca M A; Nakouzi-Naranjo, Antonio; Zuzarte, Mónica; Chatterjee, Subhasish; Stark, Ruth E; Casadevall, Arturo; Gonçalves, Teresa

2015-12-28

The importance of Alternaria species fungi to human health ranges from their role as etiological agents of serious infections with poor prognoses in immunosuppressed individuals to their association with respiratory allergic diseases. The present work focuses on Alternaria infectoria, which was used as a model organism of the genus, and was designed to unravel melanin production in response to antifungals. After we characterized the pigment produced by A. infectoria, we studied the dynamics of 1,8-dihydroxynaphthalene (DHN)-melanin production during growth, the degree of melanization in response to antifungals, and how melanization affected susceptibility to several classes of therapeutic drugs. We demonstrate that A. infectoria increased melanin deposition in cell walls in response to nikkomycin Z, caspofungin, and itraconazole but not in response to fluconazole or amphotericin B. These results indicate that A. infectoria activates DHN-melanin synthesis in response to certain antifungal drugs, possibly as a protective mechanism against these drugs. Inhibition of DHN-melanin synthesis by pyroquilon resulted in a lower minimum effective concentration (MEC) of caspofungin and enhanced morphological changes (increased hyphal balloon size), characterized by thinner and less organized A. infectoria cell walls. In summary, A. infectoria synthesizes melanin in response to certain antifungal drugs, and its susceptibility is influenced by melanization, suggesting the therapeutic potential of drug combinations that affect melanin synthesis. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Activation of Melanin Synthesis in Alternaria infectoria by Antifungal Drugs

PubMed Central

Fernandes, Chantal; Prados-Rosales, Rafael; Silva, Branca M. A.; Nakouzi-Naranjo, Antonio; Zuzarte, Mónica; Chatterjee, Subhasish; Stark, Ruth E.; Casadevall, Arturo

2015-01-01

The importance of Alternaria species fungi to human health ranges from their role as etiological agents of serious infections with poor prognoses in immunosuppressed individuals to their association with respiratory allergic diseases. The present work focuses on Alternaria infectoria, which was used as a model organism of the genus, and was designed to unravel melanin production in response to antifungals. After we characterized the pigment produced by A. infectoria, we studied the dynamics of 1,8-dihydroxynaphthalene (DHN)-melanin production during growth, the degree of melanization in response to antifungals, and how melanization affected susceptibility to several classes of therapeutic drugs. We demonstrate that A. infectoria increased melanin deposition in cell walls in response to nikkomycin Z, caspofungin, and itraconazole but not in response to fluconazole or amphotericin B. These results indicate that A. infectoria activates DHN-melanin synthesis in response to certain antifungal drugs, possibly as a protective mechanism against these drugs. Inhibition of DHN-melanin synthesis by pyroquilon resulted in a lower minimum effective concentration (MEC) of caspofungin and enhanced morphological changes (increased hyphal balloon size), characterized by thinner and less organized A. infectoria cell walls. In summary, A. infectoria synthesizes melanin in response to certain antifungal drugs, and its susceptibility is influenced by melanization, suggesting the therapeutic potential of drug combinations that affect melanin synthesis. PMID:26711773

7 CFR 93.5 - Fees for citrus product analyses set by cooperative agreement.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 3 2010-01-01 2010-01-01 false Fees for citrus product analyses set by cooperative... Citrus Products § 93.5 Fees for citrus product analyses set by cooperative agreement. The fees for the analyses of fresh citrus juices and other citrus products shall be set by mutual agreement between the...

Necrotrophic pathogens use the salicylic acid signaling pathway to promote disease development in tomato.

PubMed

Rahman, Taha Abd El; Oirdi, Mohamed El; Gonzalez-Lamothe, Rocio; Bouarab, Kamal

2012-12-01

Plants use different immune pathways to combat pathogens. The activation of the jasmonic acid (JA)-signaling pathway is required for resistance against necrotrophic pathogens; however, to combat biotrophic pathogens, the plants activate mainly the salicylic acid (SA)-signaling pathway. SA can antagonize JA signaling and vice versa. NPR1 (noninducible pathogenesis-related 1) is considered a master regulator of SA signaling. NPR1 interacts with TGA transcription factors, ultimately leading to the activation of SA-dependent responses. SA has been shown to promote disease development caused by the necrotrophic pathogen Botrytis cinerea through NPR1, by suppressing the expression of two JA-dependent defense genes, proteinase inhibitors I and II. We show here that the transcription factor TGA1.a contributes to disease development caused by B. cinerea in tomato by suppressing the expression of proteinase inhibitors I and II. Finally, we present evidence that the SA-signaling pathway contributes to disease development caused by another necrotrophic pathogen, Alternaria solani, in tomato. Disease development promoted by SA through NPR1 requires the TGA1.a transcription factor. These data highlight how necrotrophs manipulate the SAsignaling pathway to promote their disease in tomato.

Metabolomic analysis of primary metabolites in citrus leaf during defense responses.

PubMed

Asai, Tomonori; Matsukawa, Tetsuya; Kajiyama, Shin'ichiro

2017-03-01

Mechanical damage is one of the unavoidable environmental stresses to plant growth and development. Plants induce a variety of reactions which defend against natural enemies and/or heal the wounded sites. Jasmonic acid (JA) and salicylic acid (SA), defense-related plant hormones, are well known to be involved in induction of defense reactions and play important roles as signal molecules. However, defense related metabolites are so numerous and diverse that roles of individual compounds are still to be elucidated. In this report, we carried out a comprehensive analysis of metabolic changes during wound response in citrus plants which are one of the most commercially important fruit tree families. Changes in amino acid, sugar, and organic acid profiles in leaves were surveyed after wounding, JA and SA treatments using gas chromatography-mass spectrometry (GC/MS) in seven citrus species, Citrus sinensis, Citrus limon, Citrus paradisi, Citrus unshiu, Citrus kinokuni, Citrus grandis, and Citrus hassaku. GC/MS data were applied to multivariate analyses including hierarchical cluster analysis (HCA), primary component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) to extract stress-related compounds. HCA showed the amino acid cluster including phenylalanine and tryptophan, suggesting that amino acids in this cluster are concertedly regulated during responses against treatments. OPLS-DA exhibited that tryptophan was accumulated after wounding and JA treatments in all species tested, while serine was down regulated. Our results suggest that tryptophan and serine are common biomarker candidates in citrus plants for wound stress. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Molecular analysis of citrus rust (Phyllocoptruta oleivora) populations in Texas

USDA-ARS?s Scientific Manuscript database

Phyllocoptruta olevoria (Acari: Eriophyidae) or the citrus rust mite (CRM) is one of the most economically important mite pests of citrus worldwide. CRM originated from Asia, however it is found in most citrus producing areas, such as, Brazil, Morocco and the United States. Though CRM is a major pes...

Wavelength and polarization affect phototaxis of the Asian citrus psyllid

USDA-ARS?s Scientific Manuscript database

The Asian citrus psyllid, D. citri, is a primary pest for citrus production due to its status as a vector of the citrus disease, huanglongbing. We investigated phototactic behavior of D. citri to evaluate effects of light of specific wavelength or polarization using a horizontal bioassay arena. Wave...

7 CFR 319.56-59 - Fresh citrus fruit from Uruguay.

Code of Federal Regulations, 2014 CFR

2014-01-01

... Fresh citrus fruit from Uruguay. Sweet oranges (Citrus sinensis (L.) Osbeck), lemons (C. limon (L.) Burm... in accordance with labeled instructions, and waxed. (g) Treatment. (1) Citrus fruit other than lemons... of this chapter for A. fraterculus and C. capitata. (2)(i) Lemons may be shipped without a treatment...

Current Situation of Citrus Huanglongbing in Guangdong, P. R. China

USDA-ARS?s Scientific Manuscript database

Guangdong Province is an important citrus production region in China. Citrus Huanglongbing (HLB, yellow shoot disease) was observed in Guangdong probably in the late 1800’s and the disease was first studied there. Since the 1990’s, citrus production in Guangdong has gradually shifted from the coasta...

Enhanced resistance to citrus canker in transgenic mandarin expressing Xa21 from rice.

PubMed

Omar, Ahmad A; Murata, Mayara M; El-Shamy, Hesham A; Graham, James H; Grosser, Jude W

2018-04-01

Genetic engineering approaches offer an alternative method to the conventional breeding of Citrus sp. 'W. Murcott' mandarin (a hybrid of 'Murcott' and an unknown pollen parent) is one of the most commercially important cultivars grown in many regions around the world. Transformation of 'W. Murcott' mandarin was achieved by direct DNA uptake using a protoplast transformation system. DNA construct (pAO3), encoding Green Fluorescent Protein (GFP) and the cDNA of Xa21, a Xanthomonas resistance gene from rice, was used to transform protoplasts of 'W. Murcott' mandarin. Following citrus protoplast culture and regeneration, transformed micro calli were microscopically designated via GFP expression, physically isolated from non-transformed tissue, and cultured on somatic embryogenesis induction medium. More than 150 transgenic embryos were recovered and from them, ten transgenic lines were regenerated and cultured on rooting medium for shoot elongation. Transgenic shoots were micrografted and established in the greenhouse with 3-5 replicates per line. The insertion of Xa21 and GFP was confirmed by PCR and southern blot analysis. GFP expression was verified by fluorescence microscopy and western blot analysis revealed expression of Xa21 although it was variable among transgenic lines, as shown by RT-qPCR. Transgenic plants challenged with the citrus canker pathogen by syringe inoculation showed a reduction in lesion number and bacterial populations within lesions compared to non-transgenic control plants. Transgenic 'W. Murcott' mandarin lines with improved canker resistance via protoplast transformation from embryogenic callus with the Xa21 gene from rice are being evaluated under field conditions to validate the level of resistance.

Control of Citrus Huanglongbing via Trunk Injection of Plant Defense Activators and Antibiotics.

PubMed

Hu, J; Jiang, J; Wang, N

2018-02-01

Citrus huanglongbing (HLB) or greening is a devastating disease of citrus worldwide and no effective control measure is currently available. Plant defense activators environmentally friendly compounds capable of inducing resistance against many plant pathogens. Earlier studies showed that foliar spray of plant defense inducers could slow down HLB disease progress. In this study, eight plant defense activators and three antibiotics were evaluated in three field trials for their effect to control HLB by trunk injection of young and mature sweet orange trees. Results showed that four trunk injections of several activators, including salicylic acid, oxalic acid, acibenzolar-S-methyl, and potassium phosphate, provided significant control of HLB by suppressing 'Candidatus Liberibacter asiaticus' titer and disease progress. Trunk injection of penicillin, streptomycin, and oxytetracycline hydrochloride resulted in excellent control of HLB. In general, antibiotics were more effective in reduction of 'Ca. L. asiaticus' titer and HLB symptom expressions than plant defense activators. These treatments also resulted in increased yield and better fruit quality. Injection of both salicylic acid and acibenzolar-S-methyl led to significant induction of pathogenesis-related (PR) genes PR-1 and PR-2 genes. Meanwhile, injection of either potassium phosphate or oxalic acid resulted in significant induction of PR-2 or PR-15 gene expression, respectively. These results suggested that HLB diseased trees remained inducible for systemic acquired resistance under field conditions. In summary, this study presents information regarding controlling HLB via trunk injection of plant defense activators and antibiotics, which helps citrus growers in decision making regarding developing an effective HLB management program.

Chemical modification of citrus pectin: Structural, physical and rheologial implications.

PubMed

Fracasso, Aline Francielle; Perussello, Camila Augusto; Carpiné, Danielle; Petkowicz, Carmen Lúcia de Oliveira; Haminiuk, Charles Windson Isidoro

2018-04-01

The present study aimed to investigate the physical, structural and rheological modifications caused by the chemical modification process of citrus pectin. Therefore, three commercial citrus pectins with different degree of esterification were chemically modified by sequential alkali and acidic hydrolytic process to produce modified citrus pectins (MCP) with special properties. The molar mass (M w ), degree of esterification (DE), monosaccharide composition, 13 C NMR spectra, homogeneity, morphology (SEM) and rheological behavior of both native and modified citrus pectins (MCP) were investigated. The chemical modification reduced the acid uronic content (up to 28.3%) and molar mass (up to 29.98%), however, showed little influence on the degree of esterification of native pectins. Modified citrus pectins presented higher amounts of neutral monosaccharides, mainly galactose, arabinose and rhamnose, typical of the Ramnogalacturonana-I (RG-I) region. Rheological tests indicated that the native and modified citrus pectins presented pseudoplastic behavior, however, the MCP samples were less viscous, compared to the native ones. Modified samples presented better dissolution in water and less strong gels, with good stability during oscillatory shearing at 25°C. This study aims to better understand the implications that chemical modifications may impose on the structure of citrus pectins. Copyright © 2017 Elsevier B.V. All rights reserved.

Combination of UV-C treatment and Metschnikowia pulcherrimas for controlling Alternaria rot in postharvest winter jujube fruit.

PubMed

Guo, Dongqi; Zhu, Lixia; Hou, Xujie

2015-01-01

The potential of using antagonistic yeast Metschnikowia pulcherrimas alone or in combination with ultraviolet-C (UV-C) treatment for controlling Alternaria rot of winter jujube, and its effects on postharvest quality of fruit was investigated. The results showed that spore germination of Alternaria alternata was significantly inhibited by each of the 3 doses (1, 5, and 10 kJ m(-2) ) in vitro. In vivo, UV-C treatment (5 kJ m(-2) ) or antagonist yeast was capable of reducing the percentage of infected wounds and lesion diameter in artificially inoculated jujube fruits, however, in fruit treated with combination of UV-C treatment and M. pulcherrima, the percentage of infected wounds and lesion diameter was only 16.0% and 0.60 cm, respectively. The decay incidence on winter jujube fruits treated with the combination of UV-C treatment and M. pulcherrima was 23% after storage at 0 ± 1 °C for 45 d followed by 22 °C for 7 d. None of the treatments impaired quality parameters of jujube fruit. Thus, the combination of UV-C radiation and M. pulcherrima could be an alternative to synthetic fungicides for controlling postharvest Alternaria rot of winter jujube. © 2014 Institute of Food Technologists®

A genomic approach to the understanding of Xylella fastidiosa pathogenicity.

PubMed

Lambais, M R; Goldman, M H; Camargo, L E; Goldman, G H

2000-10-01

Xylella fastidiosa is a fastidious, xylem-limited bacterium that causes several economically important plant diseases, including citrus variegated chlorosis (CVC). X. fastidiosa is the first plant pathogen to have its genome completely sequenced. In addition, it is probably the least previously studied of any organism for which the complete genome sequence is available. Several pathogenicity-related genes have been identified in the X. fastidiosa genome by similarity with other bacterial genes involved in pathogenesis in plants, as well as in animals. The X. fastidiosa genome encodes different classes of proteins directly or indirectly involved in cell-cell interactions, degradation of plant cell walls, iron homeostasis, anti-oxidant responses, synthesis of toxins, and regulation of pathogenicity. Neither genes encoding members of the type III protein secretion system nor avirulence-like genes have been identified in X. fastidiosa.

Identification and characterization of Citrus yellow vein clearing virus, a putative new member of the genus Mandarivirus infecting Citrus spp.

USDA-ARS?s Scientific Manuscript database

Yellow vein clearing virus, an uncharacterized filamentous virus, was first observed in Pakistan in 1988 and later in India in 1997 in Etrog citron (Citrus medica). Based on electron microscopic evidence of filamentous particles, the virus, provisionally named Citrus yellow vein clearing virus (CYVC...

Biological Control of Alternaria Fruit Rot of Chili by Trichoderma Species under Field Conditions

PubMed Central

Rahman, M. A.; Alam, M. Firoz

2010-01-01

Trichoderma strains were evaluated under field conditions to assay their efficacy in suppressing Alternaria fruit rot disease and promoting chili plant growth. The experiment was conducted at the Botanical Garden, Rajshahi University, Bangladesh from July 2006 to March 2007. Application of Trichoderma harzianum IMI 392432 significantly (p = 0.05) suppressed the disease compared to Alternaria tenuis (T2) treatment and improved both growth and yield. The treatment T4 (T. harzianum IMI-392432 + A. tenuis) was most effective in reducing disease percentage (72.27%) compared to A. tenuis (T1) treatment. The highest seed germination rate (85.56%) and the highest growth and yield (12.5 g/plant) was also recorded in the same treatment (T4), followed by T5 (T. harzianum IMI-392433 000000 + A. tenuis), T6 (T. harzianum IMI-392434 +A. tenuis), T2 (T. virens IMI-392430 + A. tenuis), and T3 (T. pseudokoningii IMI-392431 +A. tenuis) treatment, while single treatment with A. tenuis significantly decreased these values. PMID:23956637

Chemical and biological comparison of the fruit extracts of Citrus wilsonii Tanaka and Citrus medica L.

PubMed

Zhao, Pan; Duan, Li; Guo, Long; Dou, Li-Li; Dong, Xin; Zhou, Ping; Li, Ping; Liu, E-Hu

2015-04-15

Citri Fructus (CF), the mature fruit of Citrus wilsonii Tanaka (CWT) or Citrus medica L. (CML), is an important citrus by-product with health promoting and nutritive properties. The present study compares the chemical and biological differences of CWT and CML. Thin layer chromatography and high performance liquid chromatography, coupled with quadrupole time-of-flight tandem mass spectrometry techniques, were employed to compare the chemical profiles of CWT and CML. A total of 25 compounds were identified and the results indicated that there were significant differences in chemical composition between the two CF species. The quantitative results obtained by HPLC coupled with diode array detector method demonstrated that naringin was present in the highest amounts in CWT, whilst nomilin was the most dominant constituent in CML. It was also found that CWT had significantly higher free radical-scavenging activity than CML. Copyright © 2014 Elsevier Ltd. All rights reserved.

Winter-injury following horticultural treatments to overcome juvenility in citrus seedlings

USDA-ARS?s Scientific Manuscript database

Citrus seedling juvenility delays new hybrid evaluation, slows cultivar release, and slows introgression of new traits. A horticultural program reported to overcome citrus juvenility was tested at the Whitmore Citrus Research Foundation farm (Lake County), using replicated Hirado Buntan x Clementine...

Detection of Citrus Trees from Uav Dsms

NASA Astrophysics Data System (ADS)

Ok, A. O.; Ozdarici-Ok, A.

2017-05-01

This paper presents an automated approach to detect citrus trees from digitals surface models (DSMs) as a single source. The DSMs in this study are generated from Unmanned Aerial Vehicles (UAVs), and the proposed approach first considers the symmetric nature of the citrus trees, and it computes the orientation-based radial symmetry in an efficient way. The approach also takes into account the local maxima (LM) information to verify the output of the radial symmetry. Our contributions in this study are twofold: (i) Such an integrated approach (symmetry + LM) has not been tested to detect (citrus) trees (in orchards), and (ii) the validity of such an integrated approach has not been experienced for an input, e.g. a single DSM. Experiments are performed on five test patches. The results reveal that our approach is capable of counting most of the citrus trees without manual intervention. Comparison to the state-of-the-art reveals that the proposed approach provides notable detection performance by providing the best balance between precision and recall measures.

Reinvestigation of structure of porritoxin, a phytotoxin of Alternaria porri.

PubMed

Horiuchi, Masayuki; Maoka, Takashi; Iwase, Noriyasu; Ohnishi, Keiichiro

2002-08-01

The structure of porritoxin, a phytotoxin of Alternaria porri, was reinvestigated by detailed 2D NMR analysis including (1)H-(13)C and (1)H-(15)N HMBC experiments. The structure of porritoxin was determined to be 2-(2'-hydroxyethyl)-4-methoxy-5-methyl-6-(3' '-methyl-2' '-butenyloxy)-2,3-dihydro-1H-isoindol-1-one (1). Thus our previous proposed structure, 8-(3',3'-dimethylallyloxy)-10-methoxy-9-methyl-1H-3,4-dihydro-2,5-benzoxazocin-6(5H)-one (2), is incorrect.

Precocious flowering of juvenile citrus induced by a viral vector based on Citrus leaf blotch virus: a new tool for genetics and breeding.

PubMed

Velázquez, Karelia; Agüero, Jesús; Vives, María C; Aleza, Pablo; Pina, José A; Moreno, Pedro; Navarro, Luis; Guerri, José

2016-10-01

The long juvenile period of citrus trees (often more than 6 years) has hindered genetic improvement by traditional breeding methods and genetic studies. In this work, we have developed a biotechnology tool to promote transition from the vegetative to the reproductive phase in juvenile citrus plants by expression of the Arabidopsis thaliana or citrus FLOWERING LOCUS T (FT) genes using a Citrus leaf blotch virus-based vector (clbvINpr-AtFT and clbvINpr-CiFT, respectively). Citrus plants of different genotypes graft inoculated with either of these vectors started flowering within 4-6 months, with no alteration of the plant architecture, leaf, flower or fruit morphology in comparison with noninoculated adult plants. The vector did not integrate in or recombine with the plant genome nor was it pollen or vector transmissible, albeit seed transmission at low rate was detected. The clbvINpr-AtFT is very stable, and flowering was observed over a period of at least 5 years. Precocious flowering of juvenile citrus plants after vector infection provides a helpful and safe tool to dramatically speed up genetic studies and breeding programmes. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

Risk assessment of various insecticides used for management of Asian citrus psyllid, Diaphorina citri in Florida citrus, against honey bee, Apis mellifera.

PubMed

Chen, Xue Dong; Gill, Torrence A; Pelz-Stelinski, Kirsten S; Stelinski, Lukasz L

2017-04-01

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a major pest of citrus trees worldwide. A wide variety of insecticides are used to manage D. citri populations within citrus groves in Florida. However, in areas shared by citrus growers and beekeepers the use of insecticides may increase the risks of Apis mellifera  L. (Hymenoptera: Apidae) loss and contaminated honey. The objective of this research was to determine the environmental toxicity of insecticides, spanning five different modes of action used to control D. citri, to A. mellifera. The insecticides investigated were imidacloprid, fenpropathrin, dimethoate, spinetoram and diflubenzuron. In laboratory experiments, LD 50 values were determined and ranged from 0.10 to 0.53 ng/μl for imidacloprid, fenpropathrin, dimethoate and spinetoram. LD 50 values for diflubenzuron were >1000 ng/μl. Also, a hazard quotient was determined and ranged from 1130.43 to 10893.27 for imidacloprid, fenpropathrin, dimethoate, and spinetoram. This quotient was <0.447 for diflubenzuron. In field experiments, residual activity of fenpropathrin and dimethoate applied to citrus caused significant mortality of A. mellifera 3 and 7 days after application. Spinetoram and imidacloprid were moderately toxic to A. mellifera at the recommended rates for D. citri. Diflubenzuron was not toxic to A. mellifera in the field as compared with untreated control plots. Phenoloxidase (PO) activity of A. mellifera was higher than in untreated controls when A. mellifera were exposed to 14 days old residues. The results indicate that diflubenzuron may be safe to apply in citrus when A. mellifera are foraging, while most insecticides used for management of D. citri in citrus are likely hazardous under various exposure scenarios.

Prevalence of culturable airborne spores of selected allergenic and pathogenic fungi in outdoor air

NASA Astrophysics Data System (ADS)

O'Gorman, Céline M.; Fuller, Hubert T.

2008-06-01

Temporal and spatial variations in airborne spore concentrations of selected allergenic and pathogenic fungi were examined in Dublin, Ireland, in 2005. Air samples were taken at four outdoor locations in the city every 2 weeks, coupled with measurements of meteorological conditions. Total culturable airborne fungal spore concentrations in Dublin ranged from 30-6800 colony forming units per cubic metre of air (CFU m-3) over the 12-month period. Cladosporium, Penicillium, Aspergillus and Alternaria spores were constantly present in the Dublin atmosphere, representing >20% of the total culturable spore count. Concentrations of Cladosporium increased significantly in summer and reached allergenic threshold levels, peaking at over 3200 CFU m-3 in August. Penicillium spore concentrations never reached allergenic threshold levels, with average concentrations of <150 CFU m-3. Alternaria conidia formed only 0.3% of the total culturable fungal spore count and concentrations never exceeded 50 CFU m-3, attributable to the coastal position of Dublin and its low levels of arable production. The opportunistic human pathogen Aspergillus fumigatus was present throughout the year in nominal concentrations (<10 CFU m-3), but sporadic high counts were also recorded (300-400 CFU m-3), the potential health implications of which give cause for concern. Spores of neither Cryptococcus neoformans nor Stachybotrys chartarum were detected, but airborne basidiospores of Schizophyllum commune were evidenced by the dikaryotization of monokaryon tester strains following exposure to the air. The relationships between airborne fungal spore concentrations and meteorological factors were analysed by redundancy analysis and revealed positive correlations between temperature and Cladosporium and relative humidity and Penicillium and Aspergillus.

In Vitro Activity of Glucosinolates and Their Degradation Products against Brassica-Pathogenic Bacteria and Fungi

PubMed Central

Sotelo, T.; Lema, M.; Soengas, P.; Cartea, M. E.

2014-01-01

Glucosinolates (GSLs) are secondary metabolites found in Brassica vegetables that confer on them resistance against pests and diseases. Both GSLs and glucosinolate hydrolysis products (GHPs) have shown positive effects in reducing soil pathogens. Information about their in vitro biocide effects is scarce, but previous studies have shown sinigrin GSLs and their associated allyl isothiocyanate (AITC) to be soil biocides. The objective of this work was to evaluate the biocide effects of 17 GSLs and GHPs and of leaf methanolic extracts of different GSL-enriched Brassica crops on suppressing in vitro growth of two bacterial (Xanthomonas campestris pv. campestris and Pseudomonas syringae pv. maculicola) and two fungal (Alternaria brassicae and Sclerotinia scletoriorum) Brassica pathogens. GSLs, GHPs, and methanolic leaf extracts inhibited the development of the pathogens tested compared to the control, and the effect was dose dependent. Furthermore, the biocide effects of the different compounds studied were dependent on the species and race of the pathogen. These results indicate that GSLs and their GHPs, as well as extracts of different Brassica species, have potential to inhibit pathogen growth and offer new opportunities to study the use of Brassica crops in biofumigation for the control of multiple diseases. PMID:25362058

Aldaulactone - An Original Phytotoxic Secondary Metabolite Involved in the Aggressiveness of Alternaria dauci on Carrot.

PubMed

Courtial, Julia; Hamama, Latifa; Helesbeux, Jean-Jacques; Lecomte, Mickaël; Renaux, Yann; Guichard, Esteban; Voisine, Linda; Yovanopoulos, Claire; Hamon, Bruno; Ogé, Laurent; Richomme, Pascal; Briard, Mathilde; Boureau, Tristan; Gagné, Séverine; Poupard, Pascal; Berruyer, Romain

2018-01-01

Qualitative plant resistance mechanisms and pathogen virulence have been extensively studied since the formulation of the gene-for-gene hypothesis. The mechanisms involved in the quantitative traits of aggressiveness and plant partial resistance are less well-known. Nevertheless, they are prevalent in most plant-necrotrophic pathogen interactions, including the Daucus carota - Alternaria dauci interaction. Phytotoxic metabolite production by the pathogen plays a key role in aggressiveness in these interactions. The aim of the present study was to explore the link between A. dauci aggressiveness and toxin production. We challenged carrot embryogenic cell cultures from a susceptible genotype (H1) and two partially resistant genotypes (I2 and K3) with exudates from A. dauci strains with various aggressiveness levels. Interestingly, A. dauci -resistant carrot genotypes were only affected by exudates from the most aggressive strain in our study (ITA002). Our results highlight a positive link between A. dauci aggressiveness and the fungal exudate cell toxicity. We hypothesize that the fungal exudate toxicity was linked with the amount of toxic compounds produced by the fungus. Interestingly, organic exudate production by the fungus was correlated with aggressiveness. Hence, we further analyzed the fungal organic extract using HPLC, and correlations between the observed peak intensities and fungal aggressiveness were measured. One observed peak was closely correlated with fungal aggressiveness. We succeeded in purifying this peak and NMR analysis revealed that the purified compound was a novel 10-membered benzenediol lactone, a polyketid that we named 'aldaulactone'. We used a new automated image analysis method and found that aldaulactone was toxic to in vitro cultured plant cells at those concentrations. The effects of both aldaulactone and fungal organic extracts were weaker on I2-resistant carrot cells compared to H1 carrot cells. Taken together, our results suggest

Mitigating citrus huanglongbing via effective application of antimicrobial compounds and thermotherapy

USDA-ARS?s Scientific Manuscript database

Huanglongbing (HLB) is a serious citrus disease that threatens the citrus industry worldwide. HLB is a systemic, infectious disease and the putative causal bacterium Candidatus Liberibacter asiaticus (Las) resides in citrus phloem. In this study, the effects of heat treatment, chemical formulations,...

Site Plan & Site Section of Citrus Landscape (Showing Relationship ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Site Plan & Site Section of Citrus Landscape (Showing Relationship of Groves & Irrigation System to Grove Canal) - Arlington Heights Citrus Landscape, Southwestern portion of city of Riverside, Riverside, Riverside County, CA

Citrus fruits intake and oral cancer risk: a systematic review and meta-analysis.

PubMed

Cirmi, Santa; Navarra, Michele; Woodside, Jayne V; Cantwell, Marie M

2018-05-10

To quantify the relationship between Citrus intake and risk of cancer of the oral cavity and pharynx. Systematic review and meta-analysis. Ovid MEDLINE, EMBASE, and Web of Science were searched until September 2017. Search terms included Citrus, Citrus aurantifolia, Citrus sinensis, Citrus paradisi, Citrus fruits, Citrus fruits extract, Citrus oil, fruits, oral cancer, mouth cancer, mouth neoplasm. The selection of studies and the systematic review were carried out in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. A pre-defined inclusion checklist resulted in the inclusion of articles which were (i) published in peer-reviewed scientific journals; (ii) English language; (iii) and included a measure of Citrus fruit intake and risk of oral and pharyngeal cancer. Studies were excluded if (i) preparations derived from other fruits were used, (ii) Citrus intake was combined with intake of other fruits; (iii) in vitro or animal models were used. We also excluded reviews, systematic reviews, meta-analyses, letters, personal opinions, conference abstracts and book chapters. Three reviewers independently performed the extraction of data from studies included. Seventeen studies met our inclusion criteria and were included in the final review. Pooled analyses showed that those with the highest Citrus fruit intake compared to the lowest intake had a 50% reduction in risk of oral cavity and pharyngeal cancer (OR 0.50; 95% CI 0.43-0.59). The studies included in this review and meta-analysis showed an inverse association between Citrus fruit intake and oral cancer. Copyright © 2018. Published by Elsevier Ltd.

Compatibility and Efficacy of Isaria fumosorosea with Horticultural Oils for Mitigation of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae)

PubMed Central

Ahmed, Juthi; Cave, Ronald D.; McKenzie, Cindy L.; Osborne, Lance S.

2017-01-01

Horticultural oils are an important component of integrated management programs of several phytophagous arthropods and pathogens affecting fruit, ornamentals and vegetables in greenhouse and field production systems. Although effective against the target pest, their incompatibility with biological control agents can compromise efforts to develop eco-friendly management programs for important agricultural pests. In this study, we assessed the in vitro effect of selected refined petroleum oils used in citrus and other horticultural crops with a biopesticide containing the entomopathogenic fungi, Isaria fumosorosea (PFR-97) under laboratory conditions. Further, we used leaf disk bioassays to evaluate the combined efficacy of petroleum oils and I. fumosorosea against the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae), a major pest of citrus in the United States. All five petroleum oil treatments (Orchex, Sun Pure, Conoco Blend -1, Conoco Blend -2, and JMS) were compatible with I. fumosorosea blastospores, as none of them were found to affect I. fumosorosea colony-forming units and radial fungal growth measured at 3, 6, 9, and 12 days post-inoculation. All mixed treatments performed better than I. fumosorosea alone against D. citri, where the highest mean survival time of D. citri was 12.5 ± 0.7 days. No significant differences in D. citri survival time and I. fumosorosea growth (fungal development index) on dead cadavers, which is important for determining their horizontal transmission, were observed when mixed with Orchex, Sun Pure, Conoco Blend -2, and JMS. Results indicated that horticultural oils in combination with I. fumosorosea could offer citrus growers an alternative treatment for integrating into their current management programs while battling against D. citri in citrus production systems. Due to their eco-friendly, broad-spectrum effect, it could provide control against various citrus pests, while also encouraging the retention of effective

Compatibility and Efficacy of Isaria fumosorosea with Horticultural Oils for Mitigation of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae).

PubMed

Kumar, Vivek; Avery, Pasco B; Ahmed, Juthi; Cave, Ronald D; McKenzie, Cindy L; Osborne, Lance S

2017-10-31

Horticultural oils are an important component of integrated management programs of several phytophagous arthropods and pathogens affecting fruit, ornamentals and vegetables in greenhouse and field production systems. Although effective against the target pest, their incompatibility with biological control agents can compromise efforts to develop eco-friendly management programs for important agricultural pests. In this study, we assessed the in vitro effect of selected refined petroleum oils used in citrus and other horticultural crops with a biopesticide containing the entomopathogenic fungi, Isaria fumosorosea (PFR-97) under laboratory conditions. Further, we used leaf disk bioassays to evaluate the combined efficacy of petroleum oils and I. fumosorosea against the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae), a major pest of citrus in the United States. All five petroleum oil treatments (Orchex, Sun Pure, Conoco Blend -1, Conoco Blend -2, and JMS) were compatible with I. fumosorosea blastospores, as none of them were found to affect I. fumosorosea colony-forming units and radial fungal growth measured at 3, 6, 9, and 12 days post-inoculation. All mixed treatments performed better than I. fumosorosea alone against D. citri , where the highest mean survival time of D. citri was 12.5 ± 0.7 days. No significant differences in D. citri survival time and I. fumosorosea growth (fungal development index) on dead cadavers, which is important for determining their horizontal transmission, were observed when mixed with Orchex, Sun Pure, Conoco Blend -2, and JMS. Results indicated that horticultural oils in combination with I. fumosorosea could offer citrus growers an alternative treatment for integrating into their current management programs while battling against D. citri in citrus production systems. Due to their eco-friendly, broad-spectrum effect, it could provide control against various citrus pests, while also encouraging the retention of effective

The effect of pre spring spray to reduce of citrus important pests.

PubMed

Hashemi, B; Damavandian, M R; Shoushtaril, R Vafaei; Tafaghodynia, B

2008-10-01

The importance of pre spring spray against citrus aphids, Pulvinaria aurantii Cockerell and Panonychus citri McGregor that are the most important pest of citrus during spring was tested. In this research, 150 trees ten years old sweet orange (Thomson navel on Citrus aurantium (root stocks)) in a citrus orchard approximately three hectares sampled. The experiment was laid out in a totally randomized (one-way) design replicated five times. According to the results, the pre spring spray do not effect on population density of citrus aphids and P. aurantii during March, April, May and June. However, the P. citri population decreased. Therefore, it seems the pre spring spray in citrus orchards is not necessary, but if P. citri is observed, the pre spring spray should be recommended.

Genome-wide identification of sweet orange (Citrus sinensis) histone modification gene families and their expression analysis during the fruit development and fruit-blue mold infection process

PubMed Central

Xu, Jidi; Xu, Haidan; Liu, Yuanlong; Wang, Xia; Xu, Qiang; Deng, Xiuxin

2015-01-01

In eukaryotes, histone acetylation and methylation have been known to be involved in regulating diverse developmental processes and plant defense. These histone modification events are controlled by a series of histone modification gene families. To date, there is no study regarding genome-wide characterization of histone modification related genes in citrus species. Based on the two recent sequenced sweet orange genome databases, a total of 136 CsHMs (Citrus sinensis histone modification genes), including 47 CsHMTs (histone methyltransferase genes), 23 CsHDMs (histone demethylase genes), 50 CsHATs (histone acetyltransferase genes), and 16 CsHDACs (histone deacetylase genes) were identified. These genes were categorized to 11 gene families. A comprehensive analysis of these 11 gene families was performed with chromosome locations, phylogenetic comparison, gene structures, and conserved domain compositions of proteins. In order to gain an insight into the potential roles of these genes in citrus fruit development, 42 CsHMs with high mRNA abundance in fruit tissues were selected to further analyze their expression profiles at six stages of fruit development. Interestingly, a numbers of genes were expressed highly in flesh of ripening fruit and some of them showed the increasing expression levels along with the fruit development. Furthermore, we analyzed the expression patterns of all 136 CsHMs response to the infection of blue mold (Penicillium digitatum), which is the most devastating pathogen in citrus post-harvest process. The results indicated that 20 of them showed the strong alterations of their expression levels during the fruit-pathogen infection. In conclusion, this study presents a comprehensive analysis of the histone modification gene families in sweet orange and further elucidates their behaviors during the fruit development and the blue mold infection responses. PMID:26300904

Genome-wide identification of sweet orange (Citrus sinensis) histone modification gene families and their expression analysis during the fruit development and fruit-blue mold infection process.

PubMed

Xu, Jidi; Xu, Haidan; Liu, Yuanlong; Wang, Xia; Xu, Qiang; Deng, Xiuxin

2015-01-01

In eukaryotes, histone acetylation and methylation have been known to be involved in regulating diverse developmental processes and plant defense. These histone modification events are controlled by a series of histone modification gene families. To date, there is no study regarding genome-wide characterization of histone modification related genes in citrus species. Based on the two recent sequenced sweet orange genome databases, a total of 136 CsHMs (Citrus sinensis histone modification genes), including 47 CsHMTs (histone methyltransferase genes), 23 CsHDMs (histone demethylase genes), 50 CsHATs (histone acetyltransferase genes), and 16 CsHDACs (histone deacetylase genes) were identified. These genes were categorized to 11 gene families. A comprehensive analysis of these 11 gene families was performed with chromosome locations, phylogenetic comparison, gene structures, and conserved domain compositions of proteins. In order to gain an insight into the potential roles of these genes in citrus fruit development, 42 CsHMs with high mRNA abundance in fruit tissues were selected to further analyze their expression profiles at six stages of fruit development. Interestingly, a numbers of genes were expressed highly in flesh of ripening fruit and some of them showed the increasing expression levels along with the fruit development. Furthermore, we analyzed the expression patterns of all 136 CsHMs response to the infection of blue mold (Penicillium digitatum), which is the most devastating pathogen in citrus post-harvest process. The results indicated that 20 of them showed the strong alterations of their expression levels during the fruit-pathogen infection. In conclusion, this study presents a comprehensive analysis of the histone modification gene families in sweet orange and further elucidates their behaviors during the fruit development and the blue mold infection responses.

Changes in anthocyanin production during domestication of citrus

USDA-ARS?s Scientific Manuscript database

Mandarin (C. reticulata), citron (C. medica) and pummelo (C. maxima) are imortant fruit species of the genus Citrus and parents of the interspecific hybrids that constitute the most familiar commercial varities of citrus: sweet orange, sour orange, clementine, lemon lme and grapefruit. Citron and it...

Citrus fruit quality assessment; producer and consumer perspectives

USDA-ARS?s Scientific Manuscript database

Consumption of citrus fruit and juices is popular with consumers worldwide and makes an important contribution to a healthy diet. Nevertheless, consumer preferences for citrus have undergone significant changes over the last twenty years and it is important to understand what consumers are looking ...

Monitoring Citrus Soil Moisture and Nutrients Using an IoT Based System.

PubMed

Zhang, Xueyan; Zhang, Jianwu; Li, Lin; Zhang, Yuzhu; Yang, Guocai

2017-02-23

Chongqing mountain citrus orchard is one of the main origins of Chinese citrus. Its planting terrain is complex and soil parent material is diverse. Currently, the citrus fertilization, irrigation and other management processes still have great blindness. They usually use the same pattern and the same formula rather than considering the orchard terrain features, soil differences, species characteristics and the state of tree growth. With the help of the ZigBee technology, artificial intelligence and decision support technology, this paper has developed the research on the application technology of agricultural Internet of Things for real-time monitoring of citrus soil moisture and nutrients as well as the research on the integration of fertilization and irrigation decision support system. Some achievements were obtained including single-point multi-layer citrus soil temperature and humidity detection wireless sensor nodes and citrus precision fertilization and irrigation management decision support system. They were applied in citrus base in the Three Gorges Reservoir Area. The results showed that the system could help the grower to scientifically fertilize or irrigate, improve the precision operation level of citrus production, reduce the labor cost and reduce the pollution caused by chemical fertilizer.

Consideration for alternative outlet for new citrus hybrids

USDA-ARS?s Scientific Manuscript database

Citrus sinensis (sweet orange, ex. Hamlin, Midsweet, Valencia) is the source of “orange juice” and juice must contain no less than 90% C. sinensis to be marketed as such. Juice produced from Citrus reticulata (mandarins) and C. reticulata hybrids (Orlando, Murcott, Fallglo, Sunburst, Minneola) can b...

First report of citrus leaf blotch virus in lemon in China

USDA-ARS?s Scientific Manuscript database

Citrus leaf blotch virus (CLBV) is a species of genus Citrivirus in the family Betaflexiviridae. The virus infects several species of the genus Citrus spp., but has not been previously reported from Lemon [Citrus limon (L.)]. The virus was identified in a lemon tree displaying yellow vein clearing i...

The draft genome of sweet orange (Citrus sinensis).

PubMed

Xu, Qiang; Chen, Ling-Ling; Ruan, Xiaoan; Chen, Dijun; Zhu, Andan; Chen, Chunli; Bertrand, Denis; Jiao, Wen-Biao; Hao, Bao-Hai; Lyon, Matthew P; Chen, Jiongjiong; Gao, Song; Xing, Feng; Lan, Hong; Chang, Ji-Wei; Ge, Xianhong; Lei, Yang; Hu, Qun; Miao, Yin; Wang, Lun; Xiao, Shixin; Biswas, Manosh Kumar; Zeng, Wenfang; Guo, Fei; Cao, Hongbo; Yang, Xiaoming; Xu, Xi-Wen; Cheng, Yun-Jiang; Xu, Juan; Liu, Ji-Hong; Luo, Oscar Junhong; Tang, Zhonghui; Guo, Wen-Wu; Kuang, Hanhui; Zhang, Hong-Yu; Roose, Mikeal L; Nagarajan, Niranjan; Deng, Xiu-Xin; Ruan, Yijun

2013-01-01

Oranges are an important nutritional source for human health and have immense economic value. Here we present a comprehensive analysis of the draft genome of sweet orange (Citrus sinensis). The assembled sequence covers 87.3% of the estimated orange genome, which is relatively compact, as 20% is composed of repetitive elements. We predicted 29,445 protein-coding genes, half of which are in the heterozygous state. With additional sequencing of two more citrus species and comparative analyses of seven citrus genomes, we present evidence to suggest that sweet orange originated from a backcross hybrid between pummelo and mandarin. Focused analysis on genes involved in vitamin C metabolism showed that GalUR, encoding the rate-limiting enzyme of the galacturonate pathway, is significantly upregulated in orange fruit, and the recent expansion of this gene family may provide a genomic basis. This draft genome represents a valuable resource for understanding and improving many important citrus traits in the future.

Resistance to citrus canker induced by a variant of Xanthomonas citri ssp. citri is associated with a hypersensitive cell death response involving autophagy-associated vacuolar processes.

PubMed

Roeschlin, Roxana A; Favaro, María A; Chiesa, María A; Alemano, Sergio; Vojnov, Adrián A; Castagnaro, Atilio P; Filippone, María P; Gmitter, Frederick G; Gadea, José; Marano, María R

2017-12-01

Xanthomonas citri ssp. citri (X. citri) is the causal agent of Asiatic citrus canker, a disease that seriously affects most commercially important Citrus species worldwide. We have identified previously a natural variant, X. citri A T , that triggers a host-specific defence response in Citrus limon. However, the mechanisms involved in this canker disease resistance are unknown. In this work, the defence response induced by X. citri A T was assessed by transcriptomic, physiological and ultrastructural analyses, and the effects on bacterial biofilm formation were monitored in parallel. We show that X. citri A T triggers a hypersensitive response associated with the interference of biofilm development and arrest of bacterial growth in C. limon. This plant response involves an extensive transcriptional reprogramming, setting in motion cell wall reinforcement, the oxidative burst and the accumulation of salicylic acid (SA) and phenolic compounds. Ultrastructural analyses revealed subcellular changes involving the activation of autophagy-associated vacuolar processes. Our findings show the activation of SA-dependent defence in response to X. citri A T and suggest a coordinated regulation between the SA and flavonoid pathways, which is associated with autophagy mechanisms that control pathogen invasion in C. limon. Furthermore, this defence response protects C. limon plants from disease on subsequent challenges by pathogenic X. citri. This knowledge will allow the rational exploitation of the plant immune system as a biotechnological approach for the management of the disease. © 2016 BSPP AND JOHN WILEY & SONS LTD.

Consumer preferences for fresh citrus: Impacts of demographic and behavioral characteristics

USDA-ARS?s Scientific Manuscript database

From 2000 to 2006, per capita consumption of fresh citrus fruit increased by 11.0%, but the relative shares of types of citrus consumed changed. Per capita consumption of the historically dominant citrus fruit, fresh oranges, experienced a continuous decline from 12.4 pounds to 7.4 pounds from 1990 ...

A MLVA Genotyping Scheme for Global Surveillance of the Citrus Pathogen Xanthomonas citri pv. citri Suggests a Worldwide Geographical Expansion of a Single Genetic Lineage

PubMed Central

Boyer, Karine; Leduc, Alice; Tourterel, Christophe; Drevet, Christine; Ravigné, Virginie; Gagnevin, Lionel; Guérin, Fabien; Chiroleu, Frédéric; Koebnik, Ralf; Verdier, Valérie; Vernière, Christian

2014-01-01

MultiLocus Variable number of tandem repeat Analysis (MLVA) has been extensively used to examine epidemiological and evolutionary issues on monomorphic human pathogenic bacteria, but not on bacterial plant pathogens of agricultural importance albeit such tools would improve our understanding of their epidemiology, as well as of the history of epidemics on a global scale. Xanthomonas citri pv. citri is a quarantine organism in several countries and a major threat for the citrus industry worldwide. We screened the genomes of Xanthomonas citri pv. citri strain IAPAR 306 and of phylogenetically related xanthomonads for tandem repeats. From these in silico data, an optimized MLVA scheme was developed to assess the global diversity of this monomorphic bacterium. Thirty-one minisatellite loci (MLVA-31) were selected to assess the genetic structure of 129 strains representative of the worldwide pathological and genetic diversity of X. citri pv. citri. Based on Discriminant Analysis of Principal Components (DAPC), four pathotype-specific clusters were defined. DAPC cluster 1 comprised strains that were implicated in the major geographical expansion of X. citri pv. citri during the 20th century. A subset of 12 loci (MLVA-12) resolved 89% of the total diversity and matched the genetic structure revealed by MLVA-31. MLVA-12 is proposed for routine epidemiological identification of X. citri pv. citri, whereas MLVA-31 is proposed for phylogenetic and population genetics studies. MLVA-31 represents an opportunity for international X. citri pv. citri genotyping and data sharing. The MLVA-31 data generated in this study was deposited in the Xanthomonas citri genotyping database (http://www.biopred.net/MLVA/). PMID:24897119

Early events of citrus greening (Huanglongbing) disease development at the ultrastructural level.

PubMed

Folimonova, Svetlana Y; Achor, Diann S

2010-09-01

Citrus greening (Huanglongbing [HLB]) is one of the most destructive diseases of citrus worldwide. The causal agent of HLB in Florida is thought to be 'Candidatus Liberibacter asiaticus'. Understanding of the early events in HLB infection is critical for the development of effective measures to control the disease. In this work, we conducted cytopathological studies by following the development of the disease in citrus trees graft inoculated with 'Ca. L. asiaticus'-containing material under greenhouse conditions to examine the correlation between ultrastructural changes and symptom production, with the main objective of characterizing the early events of infection. Based on our observations, one of the first degenerative changes induced upon invasion of the pathogen appears to be swelling of middle lamella between cell walls surrounding sieve elements. This anatomical aberration was often observed in samples from newly growing flushes in inoculated sweet orange and grapefruit trees at the early "presymptomatic" stage of HLB infection. Development of symptoms and their progression correlated with an increasing degree of microscopic aberrations. Remarkably, the ability to observe the bacterium in the infected tissue also correlated with the degree of the disease progression. Large numbers of bacterial cells were found in phloem sieve tubes in tissue samples from presymptomatic young flushes. In contrast, we did not observe the bacteria in highly symptomatic leaf samples, suggesting a possibility that, at more advanced stages of the disease, a major proportion of 'Ca. L. asiaticus' is present in a nonviable state. We trust that observations reported here advance our understanding of how 'Ca. L. asiaticus' causes disease. Furthermore, they may be an important aid in answering a question: when and where within an infected tree the tissue serves as a better inoculum source for acquisition and transmission of the bacterium by its psyllid vector.

An RNA-Seq-based reference transcriptome for Citrus.

PubMed

Terol, Javier; Tadeo, Francisco; Ventimilla, Daniel; Talon, Manuel

2016-03-01

Previous RNA-Seq studies in citrus have been focused on physiological processes relevant to fruit quality and productivity of the major species, especially sweet orange. Less attention has been paid to vegetative or reproductive tissues, while most Citrus species have never been analysed. In this work, we characterized the transcriptome of vegetative and reproductive tissues from 12 Citrus species from all main phylogenetic groups. Our aims were to acquire a complete view of the citrus transcriptome landscape, to improve previous functional annotations and to obtain genetic markers associated with genes of agronomic interest. 28 samples were used for RNA-Seq analysis, obtained from 12 Citrus species: C. medica, C. aurantifolia, C. limon, C. bergamia, C. clementina, C. deliciosa, C. reshni, C. maxima, C. paradisi, C. aurantium, C. sinensis and Poncirus trifoliata. Four different organs were analysed: root, phloem, leaf and flower. A total of 3421 million Illumina reads were produced and mapped against the reference C. clementina genome sequence. Transcript discovery pipeline revealed 3326 new genes, the number of genes with alternative splicing was increased to 19,739, and a total of 73,797 transcripts were identified. Differential expression studies between the four tissues showed that gene expression is overall related to the physiological function of the specific organs above any other variable. Variants discovery analysis revealed the presence of indels and SNPs in genes associated with fruit quality and productivity. Pivotal pathways in citrus such as those of flavonoids, flavonols, ethylene and auxin were also analysed in detail. © 2015 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.