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Sample records for biocontrol strain clonostachys

  1. Hydrophobins are required for conidial hydrophobicity and plant root colonization in the fungal biocontrol agent Clonostachys rosea

    PubMed Central

    2014-01-01

    Background Filamentous fungi produce small cysteine rich surface active amphiphilic hydrophobins on the outer surface of cell walls that mediate interactions between the fungus and the environment. The role of hydrophobins in surface hydrophobicity, sporulation, fruit body formation, recognition and adhesion to host surface and virulence have been reported. The aim of the present study was to characterize the biological function of hydrophobins in the fungal biocontrol agent Clonostachys rosea in order to understand their potential roles in biocontrol mechanisms. Results Based on the presence of hydrophobin domains, cysteine spacing patterns and hydropathy plots, we identified three class II hydrophobin genes in C. rosea. Gene expression analysis showed basal expression of Hyd1, Hyd2 and Hyd3 in all conditions tested with the exception of induced Hyd1 expression in conidiating mycelium. Interestingly, up-regulation of Hyd1, Hyd2 and Hyd3 was found during C. rosea self interaction compared to interactions with the fungal plant pathogens Botrytis cinerea or Fusarium graminearum in dual culture assays. Phenotypic analysis of C. rosea deletion and complementation strains showed that Hyd1 and Hyd3 are jointly required for conidial hydrophobicity, although no difference in mycelia hydrophobicity was found between wild type (WT) and mutant strains. Interestingly, mutant strains showed increased growth rates, conidiation and enhanced tolerances of conidia to abiotic stresses. Antagonism tests using in vitro dual culture and detached leaf assays showed that the mutant strains were more aggressive towards B. cinerea, F. graminearum or Rhizoctonia solani, and that aggression was partly related to earlier conidial germination and enhanced tolerance of mutant strains to secreted fungal metabolites. Furthermore, in vitro Arabidopsis thaliana root colonization assays revealed reduced root colonization ability of the ΔHyd3 strain, but not for the ΔHyd1 strain. Furthermore

  2. Draft Genome Sequence of Mycoparasite Clonostachys rosea Strain 67-1

    PubMed Central

    Sun, Zhan-Bin

    2015-01-01

    Clonostachys rosea is a promising mycoparasite. In this study, we sequenced the draft genome of the highly effective strain 67-1 using the Illumina HiSeq 2500 sequencing platform. The genome is 55.4 Mb with a G+C content of 49.2% and provides a powerful resource for future studies on the molecular mechanisms underlying Clonostachys rosea’s antagonism on fungal pathogens. PMID:26021926

  3. The ABC transporter ABCG29 is involved in H2O2 tolerance and biocontrol traits in the fungus Clonostachys rosea.

    PubMed

    Dubey, Mukesh; Jensen, Dan Funck; Karlsson, Magnus

    2016-04-01

    For successful biocontrol interactions, biological control organisms must tolerate toxic metabolites produced by themselves or plant pathogens during mycoparasitic/antagonistic interactions, by host plant during colonization of the plant, and xenobiotics present in the environment. ATP-binding cassette (ABC) transporters can play a significant role in tolerance of toxic compounds by mediating active transport across the cellular membrane. This paper reports on functional characterization of an ABC transporter ABCG29 in the biocontrol fungus Clonostachys rosea strain IK726. Gene expression analysis showed induced expression of abcG29 during exposure to the Fusarium spp. mycotoxin zearalenone (ZEA) and the fungicides Cantus, Chipco Green and Apron. Expression of abcG29 in C. rosea was significantly higher during C. rosea-C. rosea (Cr-Cr) interaction or in exposure to C. rosea culture filtrate for 2 h, compared to interaction with Fusarium graminearum or 2 h exposure to F. graminearum culture filtrate. In contrast with gene expression data, ΔabcG29 strains did not display reduced tolerance towards ZEA, fungicides or chemical agents known for inducing oxidative, cell wall or osmotic stress, compared to C. rosea WT. The exception was a significant reduction in tolerance to H2O2 (10 mM) in ΔabcG29 strains when conidia were used as an inoculum. The antagonistic ability of ΔabcG29 strains towards F. graminearum, Fusarium oxysporum or Botrytis cinerea in dual plate assays were not different compared with WT. However, in biocontrol assays ΔabcG29 strains displayed reduced ability to protect Arabidopsis thaliana leaves from B. cinerea, and barley seedling from F. graminearum as measured by an A. thaliana detached leaf assay and a barley foot rot disease assay, respectively. These data show that the ABCG29 is dispensable for ZEA and fungicides tolerance, and antagonism but not H2O2 tolerance and biocontrol effects in C. rosea.

  4. A Three-Way Transcriptomic Interaction Study of a Biocontrol Agent (Clonostachys rosea), a Fungal Pathogen (Helminthosporium solani), and a Potato Host (Solanum tuberosum).

    PubMed

    Lysøe, Erik; Dees, Merete W; Brurberg, May Bente

    2017-08-01

    Helminthosporium solani causes silver scurf, which affects the quality of potato. The biocontrol agent Clonostachys rosea greatly limited the severity of silver scurf symptoms and amount of H. solani genomic DNA in laboratory experiments. Transcriptomic analysis during interaction showed that H. solani gene expression was highly reduced when coinoculated with the biocontrol agent C. rosea, whereas gene expression of C. rosea was clearly boosted as a response to the pathogen. The most notable upregulated C. rosea genes were those encoding proteins involved in cellular response to oxidative stress, proteases, G-protein signaling, and the methyltransferase LaeA. The most notable potato response to both fungi was downregulation of defense-related genes and mitogen-activated protein kinase kinase kinases. At a later stage, this shifted, and most potato defense genes were turned on, especially those involved in terpenoid biosynthesis when H. solani was present. Some biocontrol-activated defense-related genes in potato were upregulated during early interaction with C. rosea alone that were not triggered by H. solani alone. Our results indicate that the reductions of silver scurf using C. rosea are probably due to a combination of mechanisms, including mycoparasitism, biocontrol-activated stimulation of plant defense mechanisms, microbial competition for nutrients, space, and antibiosis.

  5. Investigating the compatibility of the biocontrol agent Clonostachys rosea IK726 with prodigiosin-producing Serratia rubidaea S55 and phenazine-producing Pseudomonas chlororaphis ToZa7.

    PubMed

    Kamou, Nathalie N; Dubey, Mukesh; Tzelepis, Georgios; Menexes, Georgios; Papadakis, Emmanouil N; Karlsson, Magnus; Lagopodi, Anastasia L; Jensen, Dan Funck

    2016-05-01

    This study was carried out to assess the compatibility of the biocontrol fungus Clonostachys rosea IK726 with the phenazine-producing Pseudomonas chlororaphis ToZa7 or with the prodigiosin-producing Serratia rubidaea S55 against Fusarium oxysporum f. sp. radicis-lycopersici. The pathogen was inhibited by both strains in vitro, whereas C. rosea displayed high tolerance to S. rubidaea but not to P. chlororaphis. We hypothesized that this could be attributed to the ATP-binding cassette (ABC) proteins. The results of the reverse transcription quantitative PCR showed an induction of seven genes (abcB1, abcB20, abcB26, abcC12, abcC12, abcG8 and abcG25) from subfamilies B, C and G. In planta experiments showed a significant reduction in foot and root rot on tomato plants inoculated with C. rosea and P. chlororaphis. This study demonstrates the potential for combining different biocontrol agents and suggests an involvement of ABC transporters in secondary metabolite tolerance in C. rosea.

  6. Diversity of Clonostachys species assessed by molecular phylogenetics and MALDI-TOF mass spectrometry.

    PubMed

    Abreu, Lucas M; Moreira, Gláucia M; Ferreira, Douglas; Rodrigues-Filho, Edson; Pfenning, Ludwig H

    2014-12-01

    We assessed the species diversity among 45 strains of Clonostachys from different substrates and localities in Brazil using molecular phylogenetics, and compared the results with the phenotypic classification of strains obtained from matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Phylogenetic analyses were based on beta tubulin (Tub), ITS-LSU rDNA, and a combined Tub-ITS DNA dataset. MALDI-TOF MS analyses were performed using intact conidia and conidiophores of strains cultivated on oatmeal agar and 4% malt extract agar. Six known species were identified: Clonostachys byssicola, Clonostachys candelabrum, Clonostachys pseudochroleuca, Clonostachys rhizophaga, Clonostachys rogersoniana, and Clonostachys rosea. Two clades and two singleton lineages did not correspond to known species represented in the reference DNA dataset and were identified as Clonostachys sp. 1-4. Multivariate cluster analyses of MALDI-TOF MS data classified the strains into eight clusters and three singletons, corresponding to the ten identified species plus one additional cluster containing two strains of C. rogersoniana that split from the other co-specific strains. The consistent results of MALDI-TOF MS supported the identification of strains assigned to C. byssicola and C. pseudochroleuca, which did not form well supported clades in all phylogenetic analyses, but formed distinct clusters in the MALDI-TOF dendrograms.

  7. Discovery of secondary metabolites from Bacillus spp. biocontrol strains using genome mining and mass spectroscopy

    USDA-ARS?s Scientific Manuscript database

    Genome sequencing, data mining and mass spectrometry were used to identify secondary metabolites produced by several Bacillus spp. biocontrol strains. These biocontrol strains have shown promise in managing Fusarium head blight in wheat. Draft genomes were produced and screened in silico using genom...

  8. Complete Genome Sequence of Biocontrol Strain Pseudomonas fluorescens LBUM223

    PubMed Central

    Roquigny, Roxane; Arseneault, Tanya; Gadkar, Vijay J.; Novinscak, Amy

    2015-01-01

    Pseudomonas fluorescens LBUM223 is a plant growth-promoting rhizobacterium (PGPR) with biocontrol activity against various plant pathogens. It produces the antimicrobial metabolite phenazine-1-carboxylic acid, which is involved in the biocontrol of Streptomyces scabies, the causal agent of common scab of potato. Here, we report the complete genome sequence of P. fluorescens LBUM223. PMID:25953163

  9. Biocontrol Activity of the Local Strain of Metschnikowia pulcherrima on Different Postharvest Pathogens.

    PubMed

    Türkel, Sezai; Korukluoğlu, Mihriban; Yavuz, Mümine

    2014-01-01

    The strains of the yeast Metschnikowia pulcherrima have strong biocontrol activity against various microorganisms. Biocontrol activity of M. pulcherrima largely depends on its iron immobilizing pigment pulcherrimin. Biocontrol activity of pulcherrimin producing strain, M. pulcherrima UMY15, isolated from local vineyards, was tested on different molds that cause food spoilage. M. pulcherrima UMY15 was a very effective biocontrol agent against Penicillium roqueforti, P. italicum, P. expansum, and Aspergillus oryzae in in-vitro plate tests. However, the inhibitory activity of M. pulcherrima UMY15 was less effective on Fusarium sp. and A. niger species in biocontrol assays. In addition, M. pulcherrima UMY15 strain completely inhibited the germination and mycelia growth of A. oryzae, A. parasiticus, and Fusarium sp. spores on artificial wounds of apples when they coinoculated with M. pulcherrima UMY15. Moreover, when coinoculated, M. pulcherrima UMY15 strain also inhibited the growth of P. roqueforti, P. italicum, P. expansum, A. oryzae, Fusarium sp., and Rhizopus sp. in grape juice, indicating that M. pulcherrima UMY15 can be used as a very effective biocontrol yeast against various species of postharvest pathogens, including Penicillium, Aspergillus, Fusarium, and Rhizopus.

  10. Biocontrol Activity of the Local Strain of Metschnikowia pulcherrima on Different Postharvest Pathogens

    PubMed Central

    Türkel, Sezai; Korukluoğlu, Mihriban; Yavuz, Mümine

    2014-01-01

    The strains of the yeast Metschnikowia pulcherrima have strong biocontrol activity against various microorganisms. Biocontrol activity of M. pulcherrima largely depends on its iron immobilizing pigment pulcherrimin. Biocontrol activity of pulcherrimin producing strain, M. pulcherrima UMY15, isolated from local vineyards, was tested on different molds that cause food spoilage. M. pulcherrima UMY15 was a very effective biocontrol agent against Penicillium roqueforti, P. italicum, P. expansum, and Aspergillus oryzae in in-vitro plate tests. However, the inhibitory activity of M. pulcherrima UMY15 was less effective on Fusarium sp. and A. niger species in biocontrol assays. In addition, M. pulcherrima UMY15 strain completely inhibited the germination and mycelia growth of A. oryzae, A. parasiticus, and Fusarium sp. spores on artificial wounds of apples when they coinoculated with M. pulcherrima UMY15. Moreover, when coinoculated, M. pulcherrima UMY15 strain also inhibited the growth of P. roqueforti, P. italicum, P. expansum, A. oryzae, Fusarium sp., and Rhizopus sp. in grape juice, indicating that M. pulcherrima UMY15 can be used as a very effective biocontrol yeast against various species of postharvest pathogens, including Penicillium, Aspergillus, Fusarium, and Rhizopus. PMID:24860671

  11. Molecular-based strategies to exploit Pseudomonas biocontrol strains for environmental biotechnology applications.

    PubMed

    Mark, Genevievel; Morrissey, John P; Higgins, P; O'gara, Fergal

    2006-05-01

    Exploitation of beneficial plant-microbe interactions in the rhizosphere can result in the promotion of plant health and have significant implications for low input sustainable agriculture applications such as biocontrol. Bacteria such as Bacillus and Pseudomonas, and fungi such as Trichoderma, have been developed as commercial biocontrol products. Registration of microbial inocualants as biocontrol agents in either the European Union or the United States requires production of extensive dossiers covering efficacy, safety and risk assessment. Despite the fact that a number of Pseudomonas biocontrol products have been marketed there are still some limitations hampering the development of this technology for widespread use in agriculture. Although many strains show good performance in specific trials, this is often not translated into consistent, effective biocontrol in diverse field situations. Advances in 'Omics' technology and the publication of complete genome sequences of a number of plant-associative bacterial strains, has facilitated investigations into the molecular basis underpinning the establishment of beneficial plant-microbe interactions in the rhizosphere. The understanding of these molecular signalling processes and the functions they regulate is fundamental to promoting beneficial microbe-plant interactions, to overcome existing limitations and to designing improved strategies for the development of novel Pseudmonas biocontrol inoculant consortia.

  12. Genomic Diversity of Biocontrol Strains of Pseudomonas spp. Isolated from Aerial or Root Surfaces of Plants

    USDA-ARS?s Scientific Manuscript database

    The striking ecological, metabolic, and biochemical diversity of Pseudomonas has intrigued microbiologists for many decades. To explore the genomic diversity of biocontrol strains of Pseudomonas spp., we derived high quality draft sequences of seven strains known to suppress plant disease. The str...

  13. Genome Sequence of the Biocontrol Agent Microbacterium barkeri Strain 2011-R4

    PubMed Central

    Liu, Jian; Zhou, Qing; Ibrahim, Muhammad; Liu, He; Jin, Gulei

    2012-01-01

    Microbacterium barkeri strain 2011-R4 is a Gram-positive epiphyte which has been confirmed as a biocontrol agent against several plant pathogens in our previous studies. Here, we present the draft genome sequence of this strain, which was isolated from the rice rhizosphere in Tonglu city, Zhejiang province, China. PMID:23144410

  14. Draft Genome Sequence of a Biocontrol Rhizobacterium, Chryseobacterium kwangjuense Strain KJ1R5, Isolated from Pepper (Capsicum annuum)

    PubMed Central

    Jeong, Jin-Ju; Park, Hongjae; Park, Byeong Hyeok; Mannaa, Mohamed; Sang, Mee Kyung

    2016-01-01

    Strain KJ1R5 of the rhizobacterium Chryseobacterium kwangjuense is an effective biocontrol agent against Phytophthora blight of pepper caused by a destructive soilborne oomycete, Phytophthora capsici. Here, we present the draft genome sequence of strain KJ1R5, which contains genes related to biocontrol, plant growth promotion, and environmental stress adaptation. PMID:27103726

  15. Efficacy of water dispersible formulations of biocontrol strains of Aspergillus flavus for aflatoxin management in corn

    USDA-ARS?s Scientific Manuscript database

    Field experiments were conducted in 2011 and 2012 to evaluate the efficacy of water dispersible granule (WDG) formulations of biocontrol strains of Aspergillus flavus in controlling aflatoxin contamination of corn. In 2011, when aflatoxin was present at very high levels, no WDG treatment provided s...

  16. Characterization of a new isolate of Pseudomonas fluorescens strain Psd as a potential biocontrol agent.

    PubMed

    Upadhyay, A; Srivastava, S

    2008-08-01

    Evaluation of a new isolate of Pseudomonas fluorescens for its biocontrol properties. Strain Psd identified as Ps. fluorescens, produces secondary metabolites that are toxic to some plant-pathogenic fungi. Inhibition of fungal growth of Fusarium oxysporum and Verticillium dahliae in the presence of bacterial culture filtrate provided the first clue to its biocontrol properties. In order to determine the basis for antifungal properties, antibiotics were extracted and analysed by TLC. Both pyrrolnitrin and phenazines could be detected in the culture of Psd. Presence of response regulator gene gacA of the two component regulatory system (GacS/GacA) was established by PCR amplification and sequencing. Sequence comparison of gacA justified the taxonomic position of this strain among the known members of Pseudomonadaceae. Synthesis of other compounds like toxic lipodepsipeptide, siderophores, and HCN was also confirmed by appropriate biochemical tests. Characterization of strain Psd by various biochemical/plate tests followed by chromatographic identification of antibiotics, demonstrates its multifunctional biocontrol property. Response regulator gene gacA provides an additional genetic marker for the phylogenetic studies. Ps. fluorescens strain Psd with its multifunctional biocontrol property can be used to bioprotect the crop plants from phytopathogens.

  17. Draft Genome Sequence of Chryseobacterium sp. Strain GSE06, a Biocontrol Endophytic Bacterium Isolated from Cucumber (Cucumis sativus)

    PubMed Central

    Jeong, Jin-Ju; Park, Byeong Hyeok; Park, Hongjae

    2016-01-01

    Chryseobacterium sp. strain GSE06 is a biocontrol endophytic bacterium against the destructive soilborne oomycete Phytophthora capsici, which causes Phytophthora blight of pepper. Here, we present its draft genome sequence, which contains genes related to biocontrol traits, such as colonization, antimicrobial activity, plant growth promotion, and abiotic or biotic stress adaptation. PMID:27313310

  18. Evaluation of recycled bioplastic pellets and a sprayable formulation for application of an Aspergillus flavus biocontrol strain

    USDA-ARS?s Scientific Manuscript database

    Biocontrol of Aspergillus flavus using inoculated bioplastic granules has been proven to be effective under laboratory and field conditions. In the present study, the use of low-density pellets from recycled bioplastic as a biocontrol strain carrier was evaluated. Applying recycled bioplastic pell...

  19. Wolbachia Biocontrol Strategies for Arboviral Diseases and the Potential Influence of Resident Wolbachia Strains in Mosquitoes.

    PubMed

    Jeffries, Claire L; Walker, Thomas

    Arboviruses transmitted by mosquitoes are a major cause of human disease worldwide. The absence of vaccines and effective vector control strategies has resulted in the need for novel mosquito control strategies. The endosymbiotic bacterium Wolbachia has been proposed to form the basis for an effective mosquito biocontrol strategy. Resident strains of Wolbachia inhibit viral replication in Drosophila fruit flies and induce a reproductive phenotype known as cytoplasmic incompatibility that allows rapid invasion of insect populations. Transinfection of Wolbachia strains into the principle mosquito vector of dengue virus, Stegomyia aegypti, has resulted in dengue-refractory mosquito lines with minimal effects on mosquito fitness. Wolbachia strains have now been established in wild St. aegypti populations through open releases in dengue-endemic countries. In this review, we outline the current state of Wolbachia-based biocontrol strategies for dengue and discuss the potential impact of resident Wolbachia strains for additional target mosquito species that transmit arboviruses.

  20. The Xylella fastidiosa Biocontrol Strain EB92-1 Genome Is Very Similar and Syntenic to Pierce's Disease Strains

    PubMed Central

    Zhang, Shujian; Flores-Cruz, Zomary; Kumar, Dibyendu; Chakrabarty, Pranjib; Hopkins, Donald L.; Gabriel, Dean W.

    2011-01-01

    Xylella fastidiosa infects a wide range of plant hosts and causes economically serious diseases, including Pierce's disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 is infectious to grapevines but does not cause symptoms. The draft genome of EB92-1 reveals that it may be missing 10 potential pathogenicity effectors. PMID:21914886

  1. The Xylella fastidiosa biocontrol strain EB92-1 genome is very similar and syntenic to Pierce's disease strains.

    PubMed

    Zhang, Shujian; Flores-Cruz, Zomary; Kumar, Dibyendu; Chakrabarty, Pranjib; Hopkins, Donald L; Gabriel, Dean W

    2011-10-01

    Xylella fastidiosa infects a wide range of plant hosts and causes economically serious diseases, including Pierce's disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 is infectious to grapevines but does not cause symptoms. The draft genome of EB92-1 reveals that it may be missing 10 potential pathogenicity effectors.

  2. Draft Genome Sequence of the Pediocin-Encoding Biopreservative and Biocontrol Strain Pediococcus acidilactici D3

    PubMed Central

    Rajendran, Mahitha; Altermann, Eric

    2013-01-01

    We describe a draft genome sequence for Pediococcus acidilactici strain D3, a component of multistrain commercial cultures with biopreservative and biocontrol properties in food-based applications. Strain D3 encodes at least one antimicrobial peptide, pediocin AMPd3. The AMPd3-encoding operon exhibits high sequence similarity to the archetype pediocin, PA-1, encoded by P. acidilactici PAC 1.0. PMID:23788534

  3. Isolation of Dickeya dadantii strains from potato disease and biocontrol by their bacteriophages.

    PubMed

    Soleimani-Delfan, Abbas; Etemadifar, Zahra; Emtiazi, Giti; Bouzari, Majid

    2015-01-01

    One of the most economically important bacterial pathogens of plants and plant products is Dickeya dadantii. This bacterium causes soft rot disease in tubers and other parts of the potato and other plants of the Solanaceae family. The application of restricted host range bacteriophages as biocontrol agents has recently gained widespread interest. This study purposed to isolate the infectious agent of the potato and evaluate its biocontrol by bacteriophages. Two phytopathogenic strains were isolated from infected potatoes, identified based on biochemical and 16S rRNA gene sequencing, and submitted to GenBank as D. dadantii strain pis3 (accession no. HQ423668) and D. dadantii strain sip4 (accession no. HQ423669). Their bacteriophages were isolated from Caspian Sea water by enriching the water filtrate with D. dadantii strains as hosts using spot or overlay methods. On the basis of morphotypes, the isolated bacteriophages were identified as members of the Myoviridae and Siphoviridae families and could inhibit the growth of antibiotic resistant D. dadantii strains in culture medium. Moreover, in Dickeya infected plants treated with bacteriophage, no disease progression was detected. No significant difference was seen between phage-treated and control plants. Thus, isolated bacteriophages can be suggested for the biocontrol of plant disease caused by Dickeya strains.

  4. Isolation of Dickeya dadantii strains from potato disease and biocontrol by their bacteriophages

    PubMed Central

    Soleimani-Delfan, Abbas; Etemadifar, Zahra; Emtiazi, Giti; Bouzari, Majid

    2015-01-01

    One of the most economically important bacterial pathogens of plants and plant products is Dickeya dadantii. This bacterium causes soft rot disease in tubers and other parts of the potato and other plants of the Solanaceae family. The application of restricted host range bacteriophages as biocontrol agents has recently gained widespread interest. This study purposed to isolate the infectious agent of the potato and evaluate its biocontrol by bacteriophages. Two phytopathogenic strains were isolated from infected potatoes, identified based on biochemical and 16S rRNA gene sequencing, and submitted to GenBank as D. dadantii strain pis3 (accession no. HQ423668) and D. dadantii strain sip4 (accession no. HQ423669). Their bacteriophages were isolated from Caspian Sea water by enriching the water filtrate with D. dadantii strains as hosts using spot or overlay methods. On the basis of morphotypes, the isolated bacteriophages were identified as members of the Myoviridae and Siphoviridae families and could inhibit the growth of antibiotic resistant D. dadantii strains in culture medium. Moreover, in Dickeya infected plants treated with bacteriophage, no disease progression was detected. No significant difference was seen between phage-treated and control plants. Thus, isolated bacteriophages can be suggested for the biocontrol of plant disease caused by Dickeya strains. PMID:26413062

  5. The Global Regulator Genes from Biocontrol Strain Serratia plymuthica IC1270: Cloning, Sequencing, and Functional Studies†

    PubMed Central

    Ovadis, Marianna; Liu, Xiaoguang; Gavriel, Sagi; Ismailov, Zafar; Chet, Ilan; Chernin, Leonid

    2004-01-01

    The biocontrol activity of various fluorescent pseudomonads towards plant-pathogenic fungi is dependent upon the GacA/GacS-type two-component system of global regulators and the RpoS transcription sigma factor. In particular, these components are required for the production of antifungal antibiotics and exoenzymes. To investigate the effects of these global regulators on the expression of biocontrol factors by plant-associated bacteria other than Pseudomonas spp., gacA/gacS and rpoS homologues were cloned from biocontrol strain IC1270 of Serratia plymuthica, which produces a set of antifungal compounds, including chitinolytic enzymes and the antibiotic pyrrolnitrin. The nucleotide and deduced protein sequence alignments of the cloned gacA/gacS-like genes—tentatively designated grrA (global response regulation activator) and grrS (global response regulation sensor) and of the cloned rpoS gene revealed 64 to 93% identity with matching genes and proteins of the enteric bacteria Escherichia coli, Pectobacterium carotovora subsp. carotovora, and Serratia marcescens. grrA, grrS, and rpoS gene replacement mutants of strain IC1270 were deficient in the production of pyrrolnitrin, an exoprotease, and N-acylhomoserine lactone quorum-sensing signal molecules. However, neither mutant appeared to differ from the parental strain in the production of siderophores, and only grrA and grrS mutants were deficient in the production of a 58-kDa endochitinase, representing the involvement of other sigma factors in the regulation of strain IC1270's chitinolytic activity. Compared to the parental strain, the grrA, grrS, and rpoS mutants were markedly less capable of suppressing Rhizoctonia solani and Pythium aphanidermatum under greenhouse conditions, indicating the dependence of strain IC1270's biocontrol property on the GrrA/GrrS and RpoS global regulators. PMID:15262936

  6. Biocontrol Potential of Metchnikowia pulcherrima Strains Against Blue Mold of Apple.

    PubMed

    Janisiewicz, W J; Tworkoski, T J; Kurtzman, C P

    2001-11-01

    ABSTRACT Eight strains of Metschnikowia pulcherrima isolated over a 4-year period from an unmanaged orchard and selected for their biocontrol activity against blue mold (caused by Penicillium expansum) of apples were characterized phenotypically, genetically, and for their biocontrol potential against blue mold on apples. All strains grew well and only differed slightly in their growth in nutrient yeast dextrose broth medium at 1 degrees C after 216 h, but large differences occurred at 0 degrees C, with strain T5-A2 outgrowing other strains by more than 25% transmittance after 360 h. This strain was also one of the most resistant to diphenylamine (DPA), a postharvest antioxidant treatment. All strains required biotin for growth in minimum salt (MS) medium, although strain ST2-A10 grew slightly in MS medium containing riboflavin or folic acid, as did ST3-E1 in MS medium without vitamins. None of the strains produced killer toxins against an indicator strain of Saccharomyces cerevisiae. Analysis of Biolog data from YT plates for all eight strains using the MLCLUST program resulted in separation of the strains into one major cluster containing four strains and four scattered strains from which strain ST1-D10 was the most distant from all other strains. This was particularly apparent in 3-D and principle component analysis. Genetic differentiation of the eight strains using maximum parsimony analysis of nucleotide sequences from domain D1/D2 of nuclear large subunit (26S) ribosomal DNA resulted in detection of two clades. Strain ST1-D10 grouped with the type strain of M. pulcherrima but the remaining seven strains grouped separately, which might possibly represent a new species. All strains significantly reduced blue mold on mature Golden Delicious apples during 1 month of storage at 1 degrees C followed by 7 days at room temperature, but strains T5-A2 and T4-A2 were distinctly more effective under these conditions. Strain T5-A2 also was the most effective in tests on

  7. Characterization of biocontrol traits in the entomopathogenic nematode Heterorhabditis georgiana (Kesah strain), and phylogenetic analysis of the nematode's symbiotic bacteria.

    USDA-ARS?s Scientific Manuscript database

    Our objective was to estimate the biocontrol potential of the recently discovered entomopathogenic nematode species, Heterorhabditis georgiana (Kesha strain). Virulence and environmental tolerance were tested among several nematode species. Heterorhabditis georgiana expressed low or intermediate c...

  8. Characterization of Alcaligenes faecalis strain AD15 indicating biocontrol activity against plant pathogens.

    PubMed

    Yokoyama, Shin-ichiro; Adachi, Yoshitomi; Asakura, Shuichi; Kohyama, Erina

    2013-01-01

    Bacterial strain possessing both bacteriostatic and fungistatic activity (biocontrol activity) against pathogens of cyclamen (Cyclamen sp.) was isolated from the soil in Gifu Prefecture, Japan, and characterized with respect to its taxonomic and biocontrol properties. The sequence of its 16S rRNA gene, morphology, biochemistry, and fatty acid composition demonstrated that it is a strain most closely related to Alcaligenes faecalis subsp. faecalis LMG 1229(T). The isolate was named A. faecalis strain AD15. A. faecalis AD15 produced hydroxylamine at maximum yields of 33.3±1.7 mg/L after 16 h cultivation in LB medium and 19.0±0.44 mg/L after 19 h cultivation in synthetic medium. Moreover, minimum inhibitory concentrations of hydroxylamine against the cyclamen pathogens Pantoea agglomerans and Colletotrichum gloeosporioides were 4.20±0.98 and 16.5±0.67 mg/L. These results indicated that the biocontrol activity of strain AD15 might be attributed to hydroxylamine, a metabolite in the culture medium, and it had the potential for biopesticide application.

  9. Rhizoxin analogs contribute to the biocontrol activity of a newly isolated pseudomonas strain.

    PubMed

    Takeuchi, Kasumi; Noda, Naomi; Katayose, Yuichi; Mukai, Yoshiyuki; Numa, Hisataka; Yamada, Kosumi; Someya, Nobutaka

    2015-03-01

    Two strains of Pseudomonas sp., Os17 and St29, were newly isolated from the rhizosphere of rice and potato, respectively, by screening for 2,4-diacetylphloroglucinol producers. These strains were found to be the same species and were the closest to but different from Pseudomonas protegens among the sequenced pseudomonads, based on 16S ribosomal RNA gene and whole-genome analyses. Strain Os17 was as effective a biocontrol agent as reported for P. protegens Cab57, whereas strain St29 was less effective. The whole-genome sequences of these strains were obtained: the genomes are organized into a single circular chromosome with 6,885,464 bp, 63.5% G+C content, and 6,195 coding sequences for strain Os17; and with 6,833,117 bp, 63.3% G+C content, and 6,217 coding sequences for strain St29. Comparative genome analysis of these strains revealed that the complete rhizoxin analog biosynthesis gene cluster (approximately 79 kb) found in the Os17 genome was absent from the St29 genome. In an rzxB mutant, which lacks the polyketide synthase essential for the production of rhizoxin analogs, the growth inhibition activity against fungal and oomycete pathogens and the plant protection efficacy were attenuated compared with those of wild-type Os17. These findings suggest that rhizoxin analogs are important biocontrol factors of this strain.

  10. Small RNAs regulate the biocontrol property of fluorescent Pseudomonas strain Psd.

    PubMed

    Upadhyay, Anamika; Kochar, Mandira; Upadhyay, Ashutosh; Tripathy, Soumya; Rajam, Manchikatla Venkat; Srivastava, Sheela

    2017-03-01

    The production of biocontrol factors by Pseudomonads is reported to be controlled at the post-transcriptional level by the GacS/GacA signal transduction pathway. This involves RNA-binding translational repressor proteins, RsmA and RsmE, and the small regulatory RNAs (sRNAs) RsmX, RsmY, and RsmZ. While the former represses genes involved in secondary metabolite production, the latter relieves this repression at the end of exponential growth. We have studied the fluorescent Pseudomonas strain Psd, possessing good biocontrol potential, and confirmed the presence of rsmY and rsmZ by PCR amplification. Gene constructs for all the three small RNAs (RsmX, RsmY and RsmZ) carried on broad host-range plasmid, pME6032 were mobilized into strain Psd. Expression analysis of gacA in the recombinant strains over-expressing rsmX (Psd-pME7320), rsmY (Psd-pME6359) and rsmZ (Psd-pME6918) revealed a significant upregulation of the response regulator. Besides, a remarkable down-regulation of rsmA was also reported in all the strains. The variant strains were found to produce comparatively higher levels of phenazines. Indole acetic acid levels were higher to some extent, and strain Psd-pME6918 also showed elevated production of HCN. The tomato seedlings infected with Fusarium oxysporum and Verticillium dahliae in the presence of culture filtrate of the recombinant strains showed better plant protection response in comparison to the wild-type strain Psd. These results suggest that small RNAs are important determinants in regulation of the biocontrol property of strain Psd. Copyright © 2016 Elsevier GmbH. All rights reserved.

  11. Anthelmintic constituents of Clonostachys candelabrum.

    PubMed

    Ayers, Sloan; Zink, Deborah L; Mohn, Kenneth; Powell, Joanne S; Brown, Christine M; Bills, Gerald; Grund, Alan; Thompson, Donald; Singh, Sheo B

    2010-03-01

    Five diastereomeric polyketide glycosides, roselipins 3A-3E (1-5), have been isolated from the acetone extract of Clonostachys candelabrum on the basis of their positive anthelmintic activity. The structures of these compounds were elucidated by comparison of their NMR and MS data to those of previously reported roselipins and related structures, and were confirmed by 2D-NMR spectral analysis. Known compounds linoleic acid (6) and aurantiogliocladin (7) were also isolated as active anthelmintic components, although much less potent than the roselipins.

  12. Evolution in biocontrol strains: insight from the harlequin ladybird Harmonia axyridis

    PubMed Central

    Tayeh, Ashraf; Estoup, Arnaud; Laugier, Guillaume; Loiseau, Anne; Turgeon, Julie; Toepfer, Stefan; Facon, Benoit

    2012-01-01

    After being used as a biocontrol agent against aphids for decades without harmful consequences, the Asian harlequin ladybird Harmonia axyridis has suddenly become an invasive pest on a worldwide scale. We investigate the impact of captive breeding on several traits of this ladybird such as genetic diversity, fecundity, survival and pathogen resistance. We conducted an experiment in the laboratory to compare the fecundity and the susceptibility to the entomopathogenic fungus Beauveria bassiana of wild and biocontrol adults of H. axyridis. We compiled these new findings with already published data. Altogether, our findings suggest that mass rearing of biological control agents may strongly impact genetic diversity and life-history traits. We discuss how such changes may subsequently affect the fitness of biological control strains in natural environments. PMID:22949923

  13. Evolution in biocontrol strains: insight from the harlequin ladybird Harmonia axyridis.

    PubMed

    Tayeh, Ashraf; Estoup, Arnaud; Laugier, Guillaume; Loiseau, Anne; Turgeon, Julie; Toepfer, Stefan; Facon, Benoit

    2012-07-01

    After being used as a biocontrol agent against aphids for decades without harmful consequences, the Asian harlequin ladybird Harmonia axyridis has suddenly become an invasive pest on a worldwide scale. We investigate the impact of captive breeding on several traits of this ladybird such as genetic diversity, fecundity, survival and pathogen resistance. We conducted an experiment in the laboratory to compare the fecundity and the susceptibility to the entomopathogenic fungus Beauveria bassiana of wild and biocontrol adults of H. axyridis. We compiled these new findings with already published data. Altogether, our findings suggest that mass rearing of biological control agents may strongly impact genetic diversity and life-history traits. We discuss how such changes may subsequently affect the fitness of biological control strains in natural environments.

  14. Phenotypic and genetic characterization of Paecilomyces lilacinus strains with biocontrol activity against root-knot nematodes.

    PubMed

    Gunasekera, T S; Holland, R J; Gillings, M R; Briscoe, D A; Neethling, D C; Williams, K L; Nevalainen, K M

    2000-09-01

    Efficient selection of fungi for biological control of nematodes requires a series of screening assays. Assessment of genetic diversity in the candidate species maximizes the variety of the isolates tested and permits the assignment of a particular genotype with high nematophagous potential using a rapid novel assay. Molecular analyses also facilitate separation between isolates, allowing the identification of proprietary strains and trace biocontrol strains in the environment. The resistance of propagules to UV radiation is an important factor in the survival of a biocontrol agent. We have analyzed 15 strains of the nematophagous fungus Paecilomyces lilacinus using these principles. Arbitrarily primed DNA and allozyme assays were applied to place the isolates into genetic clusters, and demonstrated that some genetically related P. lilacinus strains exhibit widespread geographic distributions. When exposed to UV radiation, some weakly nematophagous strains were generally more susceptible than effective isolates. A microtitre tray-based assay used to screen the pathogenic activity of each isolate to Meloidogyne javanica egg masses revealed that the nematophagous ability varied between 37%-100%. However, there was no clear relationship between nematophagous ability and genetic clusters. Molecular characterizations revealed sufficient diversity to allow tracking of strains released into the environment.

  15. [Characterization of a bacterial biocontrol strain 1404 and its efficacy in controlling postharvest citrus anthracnose].

    PubMed

    Wang, Qian; Hu, Chunjin; Ke, Fanggang; Huang, Siliang; Li, Qiqin

    2010-09-01

    Anthracnose caused by Colletotrichum gloeosporioides (Penz.) Sacc. is a main disease in citrus production. To develop an effective biocontrol measure against citrus postharvest anthracnose, we screened antagonistic microbes and obtained a bacterial strain 1404 from the rhizospheric soil of chili plants in Nanning city, Guangxi, China. The objectives of the present study were to: (1) identify and characterize the antagonistic bacterium; and (2) to evaluate the efficacy of the antagonistic strain in controlling citrus postharvest anthracnose disease. Strain 1404 was identified by comparing its 16S rDNA sequence with related bacteria from GenBank database, as well as analyzing its morphological, physiological and biochemical characters. The antagonistic stability of the strain 1404 was determined by continuously transferring it on artificial media. The effect of the strain on suppressing citrus anthracnose at postharvest stage was tested by stab inoculation method. The 16S rDNA of strain 1404 was amplified with primers PF1 (5'-AGAGTTTGATCATGGCTCAG-3') and PR1 (5'-TACGGTTACCTTGTTACGACTT-3') and its sequence submitted to GenBank (accession number: GU361113). Strain 1404 clustered with the GenBank-derived Brevibacillus brevis strains in the 16S-rDNA-sequence-based phylogenetic tree at 100% bootstrap level. The morphological traits, physiological and biochemical characters of strain 1404 agreed with that of Brevibacillus brevis. Less change in the suppressive ability of antagonist against growth of Colletotrichum gloeosporioides was observed during four continuous transfers on artificial media. The average control efficacy of the strain was 64. 9 % against the disease 20 days after the antagonist application. Strain 1404 was identified as Brevibacillus brevis based on its morphological traits, phyiological and biochemical characters as well as 16S rDNA sequence analysis. The antagonist was approved to be a promising biocontrol agent. This is the first report of

  16. Characterization of the biocontrol activity of pseudomonas fluorescens strain X reveals novel genes regulated by glucose.

    PubMed

    Kremmydas, Gerasimos F; Tampakaki, Anastasia P; Georgakopoulos, Dimitrios G

    2013-01-01

    Pseudomonas fluorescens strain X, a bacterial isolate from the rhizosphere of bean seedlings, has the ability to suppress damping-off caused by the oomycete Pythium ultimum. To determine the genes controlling the biocontrol activity of strain X, transposon mutagenesis, sequencing and complementation was performed. Results indicate that, biocontrol ability of this isolate is attributed to gcd gene encoding glucose dehydrogenase, genes encoding its co-enzyme pyrroloquinoline quinone (PQQ), and two genes (sup5 and sup6) which seem to be organized in a putative operon. This operon (named supX) consists of five genes, one of which encodes a non-ribosomal peptide synthase. A unique binding site for a GntR-type transcriptional factor is localized upstream of the supX putative operon. Synteny comparison of the genes in supX revealed that they are common in the genus Pseudomonas, but with a low degree of similarity. supX shows high similarity only to the mangotoxin operon of Ps. syringae pv. syringae UMAF0158. Quantitative real-time PCR analysis indicated that transcription of supX is strongly reduced in the gcd and PQQ-minus mutants of Ps. fluorescens strain X. On the contrary, transcription of supX in the wild type is enhanced by glucose and transcription levels that appear to be higher during the stationary phase. Gcd, which uses PQQ as a cofactor, catalyses the oxidation of glucose to gluconic acid, which controls the activity of the GntR family of transcriptional factors. The genes in the supX putative operon have not been implicated before in the biocontrol of plant pathogens by pseudomonads. They are involved in the biosynthesis of an antimicrobial compound by Ps. fluorescens strain X and their transcription is controlled by glucose, possibly through the activity of a GntR-type transcriptional factor binding upstream of this putative operon.

  17. Colonization of Clonostachys rosea on soybean root inoculated with Fusarium graminearum

    USDA-ARS?s Scientific Manuscript database

    Soybean root rot, caused by Fusarium graminearum, is a devastating disease. Clonostachys rosea has been reported to have protection against plant pathogens in different crops. The objectives of this study were to determine if a strain of C. rosea (ACM941) can colonize soybean root that were inocula...

  18. [Optimization of biocontrol agent Burkholderia pyrrocinia strain JK-SH007 fermentation by response surface methodology].

    PubMed

    Li, Hao; Ren, Jiahong; Ye, Jianren

    2013-02-01

    In order to improve ferment efficiency of biocontrol agent Burkholderia pyrrocinia JK-SH007, the fermentation conditions of this strain were optimized. The optimal fermentation conditions were corn steep liquor (13.88 g/L) and glucose (3.37 g/L) by screening test, steepest ascent experiments and response surface analysis. The results showed that the cell density of JK-SH007 (1.18 x 10(9) CFU/mL) increased 1.35 times than before, and there was a 28.84% increase in antifungal activity.

  19. Endophytic colonization of olive roots by the biocontrol strain Pseudomonas fluorescens PICF7.

    PubMed

    Prieto, Pilar; Mercado-Blanco, Jesús

    2008-05-01

    Confocal microscopy combined with three-dimensional olive root tissue sectioning was used to provide evidence of the endophytic behaviour of Pseudomonas fluorescens PICF7, an effective biocontrol strain against Verticillium wilt of olive. Two derivatives of the green fluorescent protein (GFP), the enhanced green and the red fluorescent proteins, have been used to visualize simultaneously two differently fluorescently tagged populations of P. fluorescens PICF7 within olive root tissues at the single cell level. The time-course of colonization events of olive roots cv. Arbequina by strain PICF7 and the localization of tagged bacteria within olive root tissues are described. First, bacteria rapidly colonized root surfaces and were predominantly found in the differentiation zone. Thereafter, microscopy observations showed that PICF7-tagged populations eventually disappeared from the root surface, and increasingly colonized inner root tissues. Localized and limited endophytic colonization by the introduced bacteria was observed over time. Fluorescent-tagged bacteria were always visualized in the intercellular spaces of the cortex region, and no colonization of the root xylem vessels was detected at any time. To the best of our knowledge, this is the first time this approach has been used to demonstrate endophytism of a biocontrol Pseudomonas spp. strain in a woody host such as olive using a nongnotobiotic system.

  20. Strain-level diversity of secondary metabolism in the biocontrol species Aneurinibacillus migulanus.

    PubMed

    Alenezi, Faizah N; Rekik, Imen; Bełka, Marta; Ibrahim, Abrar F; Luptakova, Lenka; Jaspars, Marcel; Woodward, Steve; Belbahri, Lassaad

    2016-01-01

    Aneurinibacillus migulanus strains Nagano and NCTC 7096 show potential in biocontrol against fungal and fungus-like plant pathogens, including a wide range of Ascomycota, Basidiomycota, and Oomycetes. Differences in terms of the range of pathogens that each strain inhibits, however, suggested that production of a single antibiotic cyclic peptide, gramicidin S (GS), by the two strains, is not the sole mechanism of inhibition. The availability of four sequenced genomes of Aneurinibacillus prompted us to apply genome mining techniques to identify the bioactive potential of A. migulanus and to provide insights into the secondary metabolite arsenal of the genus Aneurinibacillus. Up to eleven secondary metabolite biosynthetic gene clusters were present in the three Aneurinibacillus species. Biosynthetic gene clusters specifying bacteriocins, microcins, non-ribosomal peptides, polyketides, terpenes, phosphonates, lasso peptides and linaridins were identified. Chitinolytic potential and iron metabolism regulation were also investigated. With increasing numbers of biocontrol bacterial genomes being sequenced and mined, the use of approaches similar to those described in this paper will lead to an increase in the numbers of environmentally friendly natural products available to use against plant diseases. Copyright © 2015 Elsevier GmbH. All rights reserved.

  1. Genotypic and phenotypic differentiation of an antifungal biocontrol strain belonging to Bacillus subtilis.

    PubMed

    Marten, P; Smalla, K; Berg, G

    2000-09-01

    Physiological and molecular fingerprints of biotechnologically relevant rhizobacteria are necessary for registration, patenting, recognition and quality checking of the strains. To characterize the biological control agent, Bacillus subtilis B2g, the strain was compared with other plant-associated B. subtilis isolates. Phenotypic characterization included biochemical and nutritional properties, in vitro activity and analysis of potential antagonistic mechanisms towards several plant pathogenic fungi. According to the phenotypic characteristics, it was not possible to differentiate the biocontrol agent from the other strains, although the enzymatic fingerprint was unique. Genotypic diversity among the isolates was characterized by molecular fingerprinting methods using REP-PCR (repetitive extragenomic palindromic PCR), and macrorestriction of genomic DNA and electrophoretic separation of DNA fragments by pulsed-field gel electrophoresis (PFGE). A protocol for PFGE analysis using restriction enzyme SfiI for B. subtilis was developed. PFGE typing of B. subtilis B2g resulted in a unique fingerprint. Therefore, it was possible to differentiate B. subtilis B2g, the biocontrol agent of Phytovit, from other antifungal B. subtilis isolates.

  2. Secondary metabolites help biocontrol strain Pseudomonas fluorescens CHA0 to escape protozoan grazing.

    PubMed

    Jousset, Alexandre; Lara, Enrique; Wall, Luis G; Valverde, Claudio

    2006-11-01

    In soil ecosystems, bacteria must cope with predation activity, which is attributed mainly to protists. The development of antipredation strategies may help bacteria maintain higher populations and persist longer in the soil. We analyzed the interaction between the root-colonizing and biocontrol strain Pseudomonas fluorescens CHA0 and three different protist isolates (an amoeba, a flagellate, and a ciliate). CHA0 produces a set of antibiotics, HCN, and an exoprotease. We observed that protists cannot grow on CHA0 but can multiply on isogenic regulatory mutants that do not produce the extracellular metabolites. The in vitro responses to CHA0 cells and its exoproducts included growth inhibition, encystation, paralysis, and cell lysis. By analyzing the responses of protists to bacterial supernatants obtained from different isogenic mutants whose production of one or more exometabolites was affected and also to culture extracts with antibiotic enrichment, we observed different contributions of the phenolic antifungal compound 2,4-diacetylphloroglucinol (DAPG) and the extracellular protease AprA to CHA0 toxicity for protists and to the encystation-reactivation cycle. The grazing pressure artificially produced by a mixture of the three protists in a microcosm system resulted in reduced colonization of cucumber roots by a regulatory isogenic CHA0 mutant unable to produce toxins. These results suggest that exometabolite production in biocontrol strain CHA0 may contribute to avoidance of protist grazing and help sustain higher populations in the rhizosphere, which may be a desirable and advantageous trait for competition with other bacteria for available resources.

  3. Biocontrol Efficacy Among Strains of Pochonia chlamydosporia Obtained from a Root-Knot Nematode Suppressive Soil

    PubMed Central

    Yang, Jiue-in; Loffredo, Angelo; Borneman, James; Becker, J. Ole

    2012-01-01

    Three Pochonia chlamydosporia var. chlamydosporia strains were isolated from a Meloidogyne incognita-suppressive soil, and then genetically characterized with multiple Pochonia-selective typing methods based on analysis of ß-tubulin, rRNA internal transcribed spacer (ITS), rRNA small subunit (SSU), and enterobacterial repetitive intergenic consensus (ERIC) PCR. All strains exhibited different patterns with the ERIC analysis. Strains 1 and 4 were similar with PCR analysis of ß-tubulin and ITS. The strains' potential as biological control agents against root-knot nematodes were examined in greenhouse trials. All three P. chlamydosporia strains significantly reduced the numbers of nematode egg masses. When chlamydospores were used as inoculum, strain 4 reduced egg numbers on tomato roots by almost 50%, and showed effects on the numbers of J2 and on nematode-caused root-galling. A newly developed SSU-based PCR analysis differentiated strain 4 from the others, and could therefore potentially be used as a screening tool for identifying other effective biocontrol strains of P. chlamydosporia var. chlamydosporia. PMID:23483846

  4. Complete Genome Sequence of Pseudomonas fluorescens LBUM636, a Strain with Biocontrol Capabilities against Late Blight of Potato

    PubMed Central

    Morrison, Christopher K.; Novinscak, Amy; Gadkar, Vijay J.; Joly, David L.

    2016-01-01

    Herein provided is the full-genome sequence of Pseudomonas fluorescens LBUM636. This strain is a plant growth-promoting rhizobacterium (PGPR) which produces phenazine-1-carboxylic acid, an antibiotic involved in the biocontrol of numerous plant pathogens, including late blight of potato caused by the plant pathogen Phytophthora infestans. PMID:27231373

  5. Reduction of aflatoxins, cyclopiazonic acid and fumonisins in corn by biocontrol strains of non-aflatoxigenic Aspergillus flavus

    USDA-ARS?s Scientific Manuscript database

    A series of field studies in corn (maize) evaluated the ability of non-aflatoxigenic biocontrol strains of Aspergillus flavus to reduce, through competitive exclusion, production in kernels of aflatoxins and cyclopiazonic acid (CPA) by A. flavus and fumonisins by Fusarium verticillioides. The abili...

  6. Systematics of the Trichoderma harzianum species complex and the re-identification of commercial biocontrol strains

    PubMed Central

    Jaklitsch, Walter; Gazis, Romina; Degenkolb, Thomas; Samuels, Gary J.

    2016-01-01

    Trichoderma harzianum is known as a cosmopolitan, ubiquitous species associated with a wide variety of substrates. It is possibly the most commonly used name in agricultural applications involving Trichoderma, including biological control of plant diseases. While various studies have suggested that T. harzianum is a species complex, only a few cryptic species are named. In the present study the taxonomy of the T. harzianum species complex is revised to include at least 14 species. Previously named species included in the complex are T. guizhouense, T. harzianum, and T. inhamatum. Two new combinations are proposed, T. lentiforme and T. lixii. Nine species are described as new, T. afarasin, T. afroharzianum, T. atrobrunneum, T. camerunense, T. endophyticum, T. neotropicale, T. pyramidale, T. rifaii and T. simmonsii. We isolated Trichoderma cultures from four commercial biocontrol products reported to contain T. harzianum. None of the biocontrol strains were identified as T. harzianum s. str. In addition, the widely applied culture ‘T. harzianum T22’ was determined to be T. afroharzianum. Some species in the T. harzianum complex appear to be exclusively endophytic, while others were only isolated from soil. Sexual states are rare. Descriptions and illustrations are provided. A secondary barcode, nuc translation elongation factor 1-α (TEF1) is needed to identify species in this complex. PMID:25661720

  7. Systematics of the Trichoderma harzianum species complex and the re-identification of commercial biocontrol strains.

    PubMed

    Chaverri, Priscila; Branco-Rocha, Fabiano; Jaklitsch, Walter; Gazis, Romina; Degenkolb, Thomas; Samuels, Gary J

    2015-01-01

    Trichoderma harzianum is known as a cosmopolitan, ubiquitous species associated with a wide variety of substrates. It is possibly the most commonly used name in agricultural applications involving Trichoderma, including biological control of plant diseases. While various studies have suggested that T. harzianum is a species complex, only a few cryptic species are named. In the present study the taxonomy of the T. harzianum species complex is revised to include at least 14 species. Previously named species included in the complex are T. guizhouense, T. harzianum, and T. inhamatum. Two new combinations are proposed, T. lentiforme and T. lixii. Nine species are described as new, T. afarasin, T. afroharzianum, T. atrobrunneum, T. camerunense, T. endophyticum, T. neotropicale, T. pyramidale, T. rifaii and T. simmonsii. We isolated Trichoderma cultures from four commercial biocontrol products reported to contain T. harzianum. None of the biocontrol strains were identified as T. harzianum s. str. In addition, the widely applied culture 'T. harzianum T22' was determined to be T. afroharzianum. Some species in the T. harzianum complex appear to be exclusively endophytic, while others were only isolated from soil. Sexual states are rare. Descriptions and illustrations are provided. A secondary barcode, nuc translation elongation factor 1-α (TEF1) is needed to identify species in this complex.

  8. Biocontrol of geosmin-producing Streptomyces spp. by two Bacillus strains from Chinese liquor.

    PubMed

    Zhi, Yan; Wu, Qun; Du, Hai; Xu, Yan

    2016-08-16

    Streptomyces spp. producing geosmin have been regarded as the most frequent and serious microbial contamination causing earthy off-flavor in Chinese liquor. It is therefore necessary to control the Streptomyces community during liquor fermentation. Biological control, using the native microbiota present in liquor making, appears to be a better solution than chemical methods. The objective of this study was to isolate native microbiota antagonistic toward Streptomyces spp. and then to evaluate the possible action mode of the antagonists. Fourteen Bacillus strains isolated from different Daqu (the fermentation starter) showed antagonistic activity against Streptomyces sampsonii, which is one of the dominant geosmin producers. Bacillus subtilis 2-16 and Bacillus amyloliquefaciens 1-45 from Maotai Daqu significantly inhibited the growth of S. sampsonii by 57.8% and 84.3% respectively, and effectively prevented the geosmin production in the simulated fermentation experiments (inoculation ratio 1:1). To probe the biocontrol mode, the ability of strain 2-16 and 1-45 to produce antimicrobial metabolites and to reduce geosmin in the fermentation system was investigated. Antimicrobial substances were identified as lipopeptides by ultra-performance liquid chromatography tandem electrospray ionization/quadrupole-time-of-flight mass spectrometry (UPLC-ESI/Q-TOF MS) and in vitro antibiotic assay. In addition, strains 2-16 and 1-45 were able to remove 45% and 15% of the geosmin respectively in the simulated solid-state fermentation. This study highlighted the potential of biocontrol, and how the use of native Bacillus species in Daqu could provide an eco-friendly method to prevent growth of Streptomyces spp. and geosmin contamination in Chinese liquor fermentation.

  9. Production of the biocontrol agent Pantoea agglomerans strain CPA-2 using commercial products and by-products.

    PubMed

    Costa, E; Teixidó, N; Usall, J; Atarés, E; Viñas, I

    2001-08-01

    The aim of this paper was to find the nitrogen and carbon sources that provide maximum biomass production of strain CPA-2 of the biocontrol agent Pantoea agglomerans and minimum cost of media, whilst maintaining biocontrol efficacy. To reduce the cost of media, commercial products and by-products were tested. P. agglomerans can be produced using a combination of nitrogen sources such as yeast extract (5 g l(-1)) and dry beer yeast (10 g l(-1)) with inexpensive carbohydrates such as sucrose (10 g l(-1)) and molasses (20 g l(-1)), respectively, maintaining the efficacy of the biocontrol agent against Penicillium digitatum and P. italicum on oranges. The results obtained in this study could be used to provide a reliable basis for a scale-up of this fermentation process to an industrial level.

  10. Environmental Factors Modulating Antibiotic and Siderophore Biosynthesis by Pseudomonas fluorescens Biocontrol Strains

    PubMed Central

    Duffy, Brion K.; Défago, Geneviève

    1999-01-01

    Understanding the environmental factors that regulate the biosynthesis of antimicrobial compounds by disease-suppressive strains of Pseudomonas fluorescens is an essential step toward improving the level and reliability of their biocontrol activity. We used liquid culture assays to identify several minerals and carbon sources which had a differential influence on the production of the antibiotics 2,4-diacetylphloroglucinol (PHL), pyoluteorin (PLT), and pyrrolnitrin and the siderophores salicylic acid and pyochelin by the model strain CHA0, which was isolated from a natural disease-suppressive soil in Switzerland. Production of PHL was stimulated by Zn2+, NH4Mo2+, and glucose; the precursor compound mono-acetylphloroglucinol was stimulated by the same factors as PHL. Production of PLT was stimulated by Zn2+, Co2+, and glycerol but was repressed by glucose. Pyrrolnitrin production was increased by fructose, mannitol, and a mixture of Zn2+ and NH4Mo2+. Pyochelin production was increased by Co2+, fructose, mannitol, and glucose. Interestingly, production of its precursor salicylic acid was increased by different factors, i.e., NH4Mo2+, glycerol, and glucose. The mixture of Zn2+ and NH4Mo2+ with fructose, mannitol, or glycerol further enhanced the production of PHL and PLT compared with either the minerals or the carbon sources used alone, but it did not improve siderophore production. Extending fermentation time from 2 to 5 days increased the accumulation of PLT, pyrrolnitrin, and pyochelin but not of PHL. When findings with CHA0 were extended to an ecologically and genetically diverse collection of 41 P. fluorescens biocontrol strains, the effect of certain factors was strain dependent, while others had a general effect. Stimulation of PHL by Zn2+ and glucose was strain dependent, whereas PLT production by all strains that can produce this compound was stimulated by Zn2+ and transiently repressed by glucose. Inorganic phosphate reduced PHL production by CHA0 and seven

  11. Environmental factors modulating antibiotic and siderophore biosynthesis by Pseudomonas fluorescens biocontrol strains.

    PubMed

    Duffy, B K; Défago, G

    1999-06-01

    Understanding the environmental factors that regulate the biosynthesis of antimicrobial compounds by disease-suppressive strains of Pseudomonas fluorescens is an essential step toward improving the level and reliability of their biocontrol activity. We used liquid culture assays to identify several minerals and carbon sources which had a differential influence on the production of the antibiotics 2,4-diacetylphloroglucinol (PHL), pyoluteorin (PLT), and pyrrolnitrin and the siderophores salicylic acid and pyochelin by the model strain CHA0, which was isolated from a natural disease-suppressive soil in Switzerland. Production of PHL was stimulated by Zn2+, NH4Mo2+, and glucose; the precursor compound mono-acetylphloroglucinol was stimulated by the same factors as PHL. Production of PLT was stimulated by Zn2+, Co2+, and glycerol but was repressed by glucose. Pyrrolnitrin production was increased by fructose, mannitol, and a mixture of Zn2+ and NH4Mo2+. Pyochelin production was increased by Co2+, fructose, mannitol, and glucose. Interestingly, production of its precursor salicylic acid was increased by different factors, i.e., NH4Mo2+, glycerol, and glucose. The mixture of Zn2+ and NH4Mo2+ with fructose, mannitol, or glycerol further enhanced the production of PHL and PLT compared with either the minerals or the carbon sources used alone, but it did not improve siderophore production. Extending fermentation time from 2 to 5 days increased the accumulation of PLT, pyrrolnitrin, and pyochelin but not of PHL. When findings with CHA0 were extended to an ecologically and genetically diverse collection of 41 P. fluorescens biocontrol strains, the effect of certain factors was strain dependent, while others had a general effect. Stimulation of PHL by Zn2+ and glucose was strain dependent, whereas PLT production by all strains that can produce this compound was stimulated by Zn2+ and transiently repressed by glucose. Inorganic phosphate reduced PHL production by CHA0 and seven

  12. Clonostachys rhizophaga and other fungi from chickpea debris in the Palouse region of the Pacific Northwest, USA

    USDA-ARS?s Scientific Manuscript database

    In 2003, 2008 and 2009 isolates of Clonostachys sp. were recovered from post-harvest chickpea debris. Representative isolates were identified as C. rhizophaga on the basis of 99% similarity of ß-tubulin DNA sequences to sequences of the type strain and 100% similarity to representative strains. In ...

  13. Development of species-, strain- and antibiotic biosynthesis-specific quantitative PCR assays for Pantoea agglomerans as tools for biocontrol monitoring.

    PubMed

    Braun-Kiewnick, Andrea; Lehmann, Andreas; Rezzonico, Fabio; Wend, Chris; Smits, Theo H M; Duffy, Brion

    2012-09-01

    Pantoea agglomerans is a cosmopolitan plant epiphytic bacterium that includes some of the most effective biological antagonists against the fire blight pathogen Erwinia amylovora, a major threat to pome fruit production worldwide. Strain E325 is commercially available as Bloomtime Biological™ in the USA and Canada. New quantitative PCR (qPCR) assays were developed for species- and strain -specific detection in the environment, and for detection of indigenous strains carrying the biocontrol antibacterial peptide biosynthesis gene paaA. The qPCR assays were highly specific, efficient and sensitive, detecting fewer than three cells per reaction or 700 colony forming units per flower, respectively. The qPCR assays were tested on field samples, giving first indications to the incidence of P. agglomerans E325 related strains, total P. agglomerans and pantocin A producing bacteria in commercial orchards. These assays will facilitate monitoring the environmental behavior of biocontrol P. agglomerans after orchard application for disease protection, proprietary strain-tracking, and streamlined screening for discovery of new biocontrol strains.

  14. Leaf application of a sprayable bioplastic-based formulation of biocontrol Aspergillus flavus strains for reduction of aflatoxins in corn.

    PubMed

    Accinelli, Cesare; Abbas, Hamed K; Vicari, Alberto; Shier, W Thomas

    2016-08-01

    Applying non-aflatoxin-producing Aspergillus flavus isolates to the soil has been shown to be effective in reducing aflatoxin levels in harvested crops, including peanuts, cotton and corn. The aim of this study was to evaluate the possibility of controlling aflatoxin contamination using a novel sprayable formulation consisting of a partially gelatinized starch-based bioplastic dispersion embedded with spores of biocontrol A. flavus strains, which is applied to the leaf surfaces of corn plants. The formulation was shown to be adherent, resulting in colonization of leaf surfaces with the biocontrol strain of A. flavus, and to reduce aflatoxin contamination of harvested kernels by up to 80% in Northern Italy and by up to 89% in the Mississippi Delta. The percentage of aflatoxin-producing isolates in the soil reservoir under leaf-treated corn was not significantly changed, even when the soil was amended with additional A. flavus as a model of changes to the soil reservoir that occur in no-till agriculture. This study indicated that it is not necessary to treat the soil reservoir in order to achieve effective biocontrol of aflatoxin contamination in kernel corn. Spraying this novel bioplastic-based formulation to leaves can be an effective alternative in the biocontrol of A. flavus in corn. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  15. Biocontrol potential and polyphasic characterization of novel native Trichoderma strains against Macrophomina phaseolina isolated from sorghum and common bean.

    PubMed

    Larralde-Corona, C P; Santiago-Mena, M R; Sifuentes-Rincón, A M; Rodríguez-Luna, I C; Rodríguez-Pérez, M A; Shirai, K; Narváez-Zapata, J A

    2008-08-01

    Native strains of Trichoderma isolated from sorghum and common bean crop soils were investigated to assess their biocontrol potential over the phytopathogenic fungus Macrophomina phaseolina, isolated from diseased plants. The Trichoderma strains were characterized with a polyphasic approach, which combined the analysis of their morphological characteristics, enzymatic activity, macro- and microculture test results, rDNA restriction patterns (AFLP), ITS1-5.8S-ITS2 rDNA sequences, and protein profiles. The integration of these data sets can be used to select new isolates as biological control agents against native fungal phytopathogens. In general, we observed a positive correlation between the secretion of beta-1,3-glucanase and N-acetylhexosaminidase, and the biocontrol capacities of all the Trichoderma isolates. Strains with the best hyperparasitic behavior against M. phaseolina isolated from diseased bean and sorghum were Trichoderma sp. (TCBG-2) and Trichoderma koningiopsis (TCBG-8), respectively.

  16. Novel Lectin-Like Bacteriocins of Biocontrol Strain Pseudomonas fluorescens Pf-5

    PubMed Central

    Parret, Annabel H. A.; Temmerman, Koen; De Mot, René

    2005-01-01

    Bacteriocin LlpA, produced by Pseudomonas sp. strain BW11M1, is a peculiar antibacterial protein due to its homology to mannose-binding lectins mostly found in monocots (A. H. A. Parret, G. Schoofs, P. Proost, and R. De Mot, J. Bacteriol. 185:897-908, 2003). Biocontrol strain Pseudomonas fluorescens Pf-5 contains two llpA-like genes, named llpA1Pf-5 and llpA2Pf-5. Recombinant Escherichia coli cells expressing llpA1Pf-5 or llpA2Pf-5 acquired bacteriocin activity and secreted a 31-kDa protein cross-reacting with LlpABW11M1 antibodies. Antibacterial activity of the recombinant proteins was evidenced by gel overlay assays. Analysis of the antimicrobial spectrum indicated that LlpA1Pf-5 and LlpA2Pf-5 are able to inhibit P. fluorescens strains, as well as the related mushroom pathogen Pseudomonas tolaasii. LlpA-type bacteriocins are characterized by a domain structure consisting of tandem monocot mannose-binding lectin (MMBL) domains. Molecular phylogeny of these MMBL domains suggests that the individual MMBL domains within an LlpA protein have evolved separately toward a specific, as yet unknown, function or, alternatively, were acquired from different ancestral sources. Our observations are consistent with earlier observations, which hinted that MMBL-like bacteriocins represent a new family of antibacterial proteins, probably with a novel mode of action. PMID:16151105

  17. The global regulator ANR is essential for Pseudomonas chlororaphis strain PA23 biocontrol.

    PubMed

    Nandi, Munmun; Selin, Carrie; Brawerman, Gabriel; Fernando, W G Dilantha; de Kievit, Teresa R

    2016-12-01

    Pseudomonas chlororaphis PA23 is a biocontrol agent capable of protecting canola from stem rot disease caused by the fungus Sclerotinia sclerotiorum. The focus of the current study was to elucidate the role of the transcriptional regulator ANR in the biocontrol capabilities of this bacterium. An anr mutant was created, PA23anr, that was devoid antifungal activity. In other pseudomonads, ANR is essential for regulating HCN production. Characterization of PA23anr revealed that, in addition to HCN, ANR controls phenazine (PHZ), pyrrolnitrin (PRN), protease and autoinducer (AHL) signal molecule production. In gene expression studies, hcnA, phzA, prnA and phzI were found to be downregulated, consistent with our endproduct analysis. Because the phenotype of PA23anr closely resembles that of quorum sensing (QS)-deficient strains, we explored whether there is a connection between ANR and the PhzRI QS system. Both phzI and phzR are positively regulated by ANR, whereas PhzR represses anr transcription. Complementation of PA23anr with pUCP-phzR, C6-HSL or both yielded no change in phenotype. Conversely, PA23phzR harbouring pUCP23-anr exhibited partial-to-full restoration of antifungal activity, HCN, PRN and AHL production together with hcnA, prnA, phzI and rpoS expression. PHZ and protease production remained unchanged indicating that ANR can complement the QS-deficient phenotype with respect to some but not all traits. Our experiments were conducted at atmospheric O2 levels underscoring the fact that ANR has a profound effect on PA23 physiology under aerobic conditions.

  18. Characterization of antagonistic-potential of two Bacillus strains and their biocontrol activity against Rhizoctonia solani in tomato.

    PubMed

    Solanki, Manoj Kumar; Singh, Rajesh Kumar; Srivastava, Supriya; Kumar, Sudheer; Kashyap, Prem Lal; Srivastava, Alok K

    2015-01-01

    To investigate the biocontrol mechanism of two antagonistic Bacillus strains (Bacillus subtilis MB14 and Bacillus amyloliquefaciens MB101), three in vitro antagonism assays were screened and the results were concluded that both strains inhibited Rhizoctonia solani growth in a similar manner by dual culture assay, but the maximum percent of inhibition only resulted with MB101 by volatile and diffusible metabolite assays. Moreover, cell free supernatant (CFS) of MB101 also showed significant (p > 0.05) growth inhibition as compared to MB14, when 10 and 20% CFS mix with the growth medium of R. solani. After in vitro-validation, both strains were evaluated under greenhouse and the results concluded that strain MB101 had significant biocontrol potential as compared to MB14. Strain MB101 was enhanced the plant height, biomass and chlorophyll content of tomato plant through a higher degree of root colonization. In field trials, strain MB101 showed higher lessening in root rot symptoms with significant fruit yield as compare to strain MB14 and infected control. Next to the field study, the presence of four antibiotic genes (srfAA, fenD, ituC, and bmyB) also concluded the antifungal nature of both Bacillus strains. Phylogenetic analysis of protein sequences revealed a close relatedness of three genes (srfAA, fenD, and ituC) with earlier reported sequences of B. subtilis and B. amyloliquefaciens. However, bmyB showed heterogeneity in among both strains (MB14 and MB101) and it may be concluded that higher degree of antagonism, root colonization and different antibiotic producing genes may play an important role in biocontrol mechanism of strain MB101.

  19. Comparative Genomics Within the Bacillus Genus Reveal the Singularities of Two Robust Bacillus amyloliquefaciens Biocontrol Strains.

    PubMed

    Magno-Pérez-Bryan, M C; Martínez-García, P M; Hierrezuelo, J; Rodríguez-Palenzuela, P; Arrebola, E; Ramos, C; de Vicente, A; Pérez-García, A; Romero, D

    2015-10-01

    Bacillus amyloliquefaciens CECT 8237 and CECT 8238, formerly known as Bacillus subtilis UMAF6639 and UMAF6614, respectively, contribute to plant health by facing microbial pathogens or inducing the plant's defense mechanisms. We sequenced their genomes and developed a set of ad hoc scripts that allowed us to search for the features implicated in their beneficial interaction with plants. We define a core set of genes that should ideally be found in any beneficial Bacillus strain, including the production of secondary metabolites, volatile compounds, metabolic plasticity, cell-to-cell communication systems, and biofilm formation. We experimentally prove that some of these genetic elements are active, such as i) the production of known secondary metabolites or ii) acetoin and 2-3-butanediol, compounds that stimulate plant growth and host defense responses. A comparison with other Bacillus genomes permits us to find differences in the cell-to-cell communication system and biofilm formation and to hypothesize variations in their persistence and resistance ability in diverse environmental conditions. In addition, the major protection provided by CECT 8237 and CECT 8238, which is different from other Bacillus strains against bacterial and fungal melon diseases, permits us to propose a correlation with their singular genetic background and determine the need to search for additional blind biocontrol-related features.

  20. Characterization of biocontrol bacterial strains isolated from a suppressiveness-induced soil after amendment with composted almond shells.

    PubMed

    Vida, Carmen; Cazorla, Francisco M; de Vicente, Antonio

    The improvement in soil quality of avocado crops through organic amendments with composted almond shells has a positive effect on crop yield and plant health, and enhances soil suppressiveness against the phytopathogenic fungus Rosellinia necatrix. In previous studies, induced soil suppressiveness against this pathogen was related to stimulation of Gammaproteobacteria, especially some members of Pseudomonas spp. with biocontrol-related activities. In this work, we isolated bacteria from this suppressiveness-induced amended soil using a selective medium for Pseudomonas-like microorganisms. We characterized the obtained bacterial collection to aid in identification, including metabolic profiles, antagonistic responses, hybridization to biosynthetic genes of antifungal compounds, production of lytic exoenzymatic activities and plant growth-promotion-related traits, and sequenced and compared amplified 16S rDNA genes from representative bacteria. The final selection of representative strains mainly belonged to the genus Pseudomonas, but also included the genera Serratia and Stenotrophomonas. Their biocontrol-related activities were assayed using the experimental avocado model, and results showed that all selected strains protected the avocado roots against R. necatrix. This work confirmed the biocontrol activity of these Gammaproteobacteria-related members against R. necatrix following specific stimulation in a suppressiveness-induced soil after a composted almond shell application. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  1. Insights on the Evolution of Mycoparasitism from the Genome of Clonostachys rosea

    PubMed Central

    Karlsson, Magnus; Durling, Mikael Brandström; Choi, Jaeyoung; Kosawang, Chatchai; Lackner, Gerald; Tzelepis, Georgios D.; Nygren, Kristiina; Dubey, Mukesh K.; Kamou, Nathalie; Levasseur, Anthony; Zapparata, Antonio; Wang, Jinhui; Amby, Daniel Buchvaldt; Jensen, Birgit; Sarrocco, Sabrina; Panteris, Emmanuel; Lagopodi, Anastasia L.; Pöggeler, Stefanie; Vannacci, Giovanni; Collinge, David B.; Hoffmeister, Dirk; Henrissat, Bernard; Lee, Yong-Hwan; Jensen, Dan Funck

    2015-01-01

    Clonostachys rosea is a mycoparasitic fungus that can control several important plant diseases. Here, we report on the genome sequencing of C. rosea and a comparative genome analysis, in order to resolve the phylogenetic placement of C. rosea and to study the evolution of mycoparasitism as a fungal lifestyle. The genome of C. rosea is estimated to 58.3 Mb, and contains 14,268 predicted genes. A phylogenomic analysis shows that C. rosea clusters as sister taxon to plant pathogenic Fusarium species, with mycoparasitic/saprotrophic Trichoderma species in an ancestral position. A comparative analysis of gene family evolution reveals several distinct differences between the included mycoparasites. Clonostachys rosea contains significantly more ATP-binding cassette (ABC) transporters, polyketide synthases, cytochrome P450 monooxygenases, pectin lyases, glucose-methanol-choline oxidoreductases, and lytic polysaccharide monooxygenases compared with other fungi in the Hypocreales. Interestingly, the increase of ABC transporter gene number in C. rosea is associated with phylogenetic subgroups B (multidrug resistance proteins) and G (pleiotropic drug resistance transporters), whereas an increase in subgroup C (multidrug resistance-associated proteins) is evident in Trichoderma virens. In contrast with mycoparasitic Trichoderma species, C. rosea contains very few chitinases. Expression of six group B and group G ABC transporter genes was induced in C. rosea during exposure to the Fusarium mycotoxin zearalenone, the fungicide Boscalid or metabolites from the biocontrol bacterium Pseudomonas chlororaphis. The data suggest that tolerance toward secondary metabolites is a prominent feature in the biology of C. rosea. PMID:25575496

  2. Comparative transcriptome analysis of the biocontrol strain Bacillus amyloliquefaciens FZB42 as response to biofilm formation analyzed by RNA sequencing.

    PubMed

    Kröber, Magdalena; Verwaaijen, Bart; Wibberg, Daniel; Winkler, Anika; Pühler, Alfred; Schlüter, Andreas

    2016-08-10

    The strain Bacillus amyloliquefaciens FZB42 is a plant growth promoting rhizobacterium (PGPR) and biocontrol agent known to keep infections of lettuce (Lactuca sativa) by the phytopathogen Rhizoctonia solani down. Several mechanisms, including the production of secondary metabolites possessing antimicrobial properties and induction of the host plant's systemic resistance (ISR), were proposed to explain the biocontrol effect of the strain. B. amyloliquefaciens FZB42 is able to form plaques (biofilm-like structures) on plant roots and this feature was discussed to be associated with its biocontrol properties. For this reason, formation of B. amyloliquefaciens biofilms was studied at the transcriptional level using high-throughput sequencing of whole transcriptome cDNA libraries from cells grown under biofilm-forming conditions vs. planktonic growth. Comparison of the transcriptional profiles of B. amyloliquefaciens FZB42 under these growth conditions revealed a common set of highly transcribed genes mostly associated with basic cellular functions. The lci gene, encoding an antimicrobial peptide (AMP), was among the most highly transcribed genes of cells under both growth conditions suggesting that AMP production may contribute to biocontrol. In contrast, gene clusters coding for synthesis of secondary metabolites with antimicrobial properties were only moderately transcribed and not induced in biofilm-forming cells. Differential gene expression revealed that 331 genes were significantly up-regulated and 230 genes were down-regulated in the transcriptome of B. amyloliquefaciens FZB42 under biofilm-forming conditions in comparison to planktonic cells. Among the most highly up-regulated genes, the yvqHI operon, coding for products involved in nisin (class I bacteriocin) resistance, was identified. In addition, an operon whose products play a role in fructosamine metabolism was enhanced in its transcription. Moreover, genes involved in the production of the extracellular

  3. Biopriming of Infected Carrot Seed with an Antagonist, Clonostachys rosea, Selected for Control of Seedborne Alternaria spp.

    PubMed

    Jensen, Birgit; Knudsen, Inge M B; Madsen, Mette; Jensen, Dan Funck

    2004-06-01

    ABSTRACT An ecological approach was used to select fungal antagonists effective against the seedborne pathogens Alternaria dauci and A. radicina on carrot. Twenty-five and 105 isolates originating from cereal and carrot habitats were screened against the pathogens in planta, respectively. Irrespective of isolate origin, fungal isolates belonging to Clonostachys rosea controlled pre- and postemergence death caused by A. dauci and A. radicina as effectively as the fungicide iprodione. Isolate IK726 of C. rosea was used in biopriming a seed lot with 29% A. radicina and 11% A. dauci (highly infected), and a seed lot with 4% A. radicina and 7% A. dauci (low infection). Seeds were primed with water alone (hydropriming) or with addition of C. rosea IK726 (biopriming). The occurrence of A. radicina and A. dauci increased twofold and fivefold, respectively, during 14 days hydropriming, irrespective of the initial infection level. On highly infected seed, biopriming reduced the incidence of A. radicina to <2.3% and that of A. dauci to <4.8% while the level of both pathogens was <0.5% on bioprimed seed with a low initial infection rate. In sand stand establishment tests, hydroprimed seeds had a lower healthy seedling stand than nonprimed seeds, mainly due to a high degree of postemergence seedling death. In contrast, biopriming resulted in a seedling stand that was better than that of both nonprimed and hydroprimed seeds. C. rosea IK726 multiplied fivefold to eightfold, and microscopic observations using C. rosea IK726 transformed with a green fluorescent protein (GFP) reporter gene showed that seeds were covered with a fine web of sporulating mycelium of C. rosea. The positive effect of biopriming on healthy seedling stand remained after 5 months of storage at 4 degrees C and IK726 survived at high numbers on these seed. In this study, we demonstrated that bio-priming with the biocontrol strain C. rosea IK726 facilitates priming of infected seeds without risking adverse

  4. Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1

    PubMed Central

    Zhang, Shujian; Chakrabarty, Pranjib K.; Fleites, Laura A.; Rayside, Patricia A.; Hopkins, Donald L.; Gabriel, Dean W.

    2015-01-01

    Xylella fastidiosa (X. fastidiosa) infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703) and a serine protease (PD0956); two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928), and at least one relatively short, hemagglutinin-like protein (PD0986). Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3) overexpressing PD1703 exhibited a hypersensitive response (HR) in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot) and a PD0986 (hemagglutinin) were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry. PMID:26218423

  5. Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1.

    PubMed

    Zhang, Shujian; Chakrabarty, Pranjib K; Fleites, Laura A; Rayside, Patricia A; Hopkins, Donald L; Gabriel, Dean W

    2015-01-01

    Xylella fastidiosa (X. fastidiosa) infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703) and a serine protease (PD0956); two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928), and at least one relatively short, hemagglutinin-like protein (PD0986). Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3) overexpressing PD1703 exhibited a hypersensitive response (HR) in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot) and a PD0986 (hemagglutinin) were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry.

  6. Rhizocompetence and antagonistic activity towards genetically diverse Ralstonia solanacearum strains--an improved strategy for selecting biocontrol agents.

    PubMed

    Xue, Qing-Yun; Ding, Guo-Chun; Li, Shi-Mo; Yang, Yang; Lan, Cheng-Zhong; Guo, Jian-Hua; Smalla, Kornelia

    2013-02-01

    Bacterial wilt caused by Ralstonia solanacearum is a serious threat for agricultural production in China. Eight soil bacterial isolates with activity against R. solanacearum TM15 (biovar 3) were tested in this study for their in vitro activity towards ten genetically diverse R. solanacearum isolates from China. The results indicated that each antagonist showed remarkable differences in its ability to in vitro antagonize the ten different R. solanacearum strains. Strain XY21 (based on 16S rRNA gene sequencing affiliated to Serratia) was selected for further studies based on its in vitro antagonistic activity and its excellent rhizocompetence on tomato plants. Under greenhouse conditions XY21 mediated biocontrol of tomato wilt caused by seven different R. solanacearum strains ranged from 19 to 70 %. The establishment of XY21 and its effects on the bacterial community in the tomato rhizosphere were monitored by denaturing gradient gel electrophoresis of 16S rRNA gene fragments PCR-amplified from total community DNA. A positive correlation of the in vitro antagonistic activities of XY21 and the actual biocontrol efficacies towards seven genetically different R. solanacearum strains was found and further confirmed by the efficacy of XY21 in controlling bacterial wilt under field conditions.

  7. Killer activity of Saccharomyces cerevisiae strains: partial characterization and strategies to improve the biocontrol efficacy in winemaking.

    PubMed

    de Ullivarri, Miguel Fernández; Mendoza, Lucía M; Raya, Raúl R

    2014-11-01

    Killer yeasts are considered potential biocontrol agents to avoid or reduce wine spoilage by undesirable species. In this study two Saccharomyces cerevisiae strains (Cf8 and M12) producing killer toxin were partially characterized and new strategies to improve their activity in winemaking were evaluated. Killer toxins were characterized by biochemical tests and growth inhibition of sensitive yeasts. Also genes encoding killer toxin were detected in the chromosomes of both strains by PCR. Both toxins showed optimal activity and production at conditions used during the wine-making process (pH 3.5 and temperatures of 15-25 °C). In addition, production of both toxins was higher when a nitrogen source was added. To improve killer activity different strategies of inoculation were studied, with the sequential inoculation of killer strains the best combination to control the growth of undesired yeasts. Sequential inoculation of Cf8-M12 showed a 45 % increase of killer activity on sensitive S. cerevisiae and spoilage yeasts. In the presence of ethanol (5-12 %) and SO2 (50 mg/L) the killer activity of both toxins was increased, especially for toxin Cf8. Characteristics of both killer strains support their future application as starter cultures and biocontrol agents to produce wines of controlled quality.

  8. Biological Control of Aflatoxin Contamination in U.S. Crops and the Use of Bioplastic Formulations of Aspergillus flavus Biocontrol Strains To Optimize Application Strategies.

    PubMed

    Abbas, Hamed K; Accinelli, Cesare; Shier, W Thomas

    2017-08-23

    Aflatoxin contamination has a major economic impact on crop production in the southern United States. Reduction of aflatoxin contamination in harvested crops has been achieved by applying nonaflatoxigenic biocontrol Aspergillus flavus strains that can out-compete wild aflatoxigenic A. flavus, reducing their numbers at the site of application. Currently, the standard method for applying biocontrol A. flavus strains to soil is using a nutrient-supplying carrier (e.g., pearled barley for Afla-Guard). Granules of Bioplastic (partially acetylated corn starch) have been investigated as an alternative nutritive carrier for biocontrol agents. Bioplastic granules have also been used to prepare a sprayable biocontrol formulation that gives effective reduction of aflatoxin contamination in harvested corn kernels with application of much smaller amounts to leaves later in the growing season. The ultimate goal of biocontrol research is to produce biocontrol systems that can be applied to crops only when long-range weather forecasting indicates they will be needed.

  9. Bacillus methylotrophicus Strain NKG-1, Isolated from Changbai Mountain, China, Has Potential Applications as a Biofertilizer or Biocontrol Agent

    PubMed Central

    Ge, Beibei; Liu, Binghua; Nwet, Thinn Thinn; Zhao, Wenjun; Shi, Liming; Zhang, Kecheng

    2016-01-01

    Chemical pesticides are widely used in agriculture, which endangers both environmental health and food safety. Biocontrol is an environmentally-friendly and cost-effective green technique in environmental protection and agricultural production; it generally uses selected bioresources, including beneficial microorganisms. We isolated a novel bacterial strain (NKG-1) from the rare dormant volcanic soils of Changbai Mountain in China’s Jilin Province. The strain was identified as Bacillus methylotrophicus using morphological, biochemical, physiological, and phylogenetic 16S rDNA sequencing data. This strain was able to suppress mycelial growth and conidial germination of numerous plant pathogenic fungi on solid media. A greenhouse experiment showed that application of NKG-1 fermentation broth prior to inoculation of Botrytis cinerea, the cause of gray tomato mold, inhibited growth of the mold by 60%. Furthermore, application of a 100× dilution of NKG-1 fermentation broth to tomato seedlings yielded a significant increase in seedling fresh weight (27.4%), seedling length (12.5%), and root length (57.7%) compared to the control. When the same dosage was applied in the field, we observed increases in tomato plant height (14.7%), stem diameter (12.7%), crown width (16.3%), and maximum fruit diameter (11.5%). These results suggest that NKG-1 has potential commercial application as a biofertilizer or biocontrol agent. PMID:27832162

  10. Bacillus methylotrophicus Strain NKG-1, Isolated from Changbai Mountain, China, Has Potential Applications as a Biofertilizer or Biocontrol Agent.

    PubMed

    Ge, Beibei; Liu, Binghua; Nwet, Thinn Thinn; Zhao, Wenjun; Shi, Liming; Zhang, Kecheng

    2016-01-01

    Chemical pesticides are widely used in agriculture, which endangers both environmental health and food safety. Biocontrol is an environmentally-friendly and cost-effective green technique in environmental protection and agricultural production; it generally uses selected bioresources, including beneficial microorganisms. We isolated a novel bacterial strain (NKG-1) from the rare dormant volcanic soils of Changbai Mountain in China's Jilin Province. The strain was identified as Bacillus methylotrophicus using morphological, biochemical, physiological, and phylogenetic 16S rDNA sequencing data. This strain was able to suppress mycelial growth and conidial germination of numerous plant pathogenic fungi on solid media. A greenhouse experiment showed that application of NKG-1 fermentation broth prior to inoculation of Botrytis cinerea, the cause of gray tomato mold, inhibited growth of the mold by 60%. Furthermore, application of a 100× dilution of NKG-1 fermentation broth to tomato seedlings yielded a significant increase in seedling fresh weight (27.4%), seedling length (12.5%), and root length (57.7%) compared to the control. When the same dosage was applied in the field, we observed increases in tomato plant height (14.7%), stem diameter (12.7%), crown width (16.3%), and maximum fruit diameter (11.5%). These results suggest that NKG-1 has potential commercial application as a biofertilizer or biocontrol agent.

  11. Draft Genome Sequence of Biocontrol Agent Pythium oligandrum Strain Po37, an Oomycota

    PubMed Central

    Berger, Harald; Yacoub, Amira; Gerbore, Jonathan; Grizard, Damien; Rey, Patrice; Sessitsch, Angela

    2016-01-01

    The oomycota Pythium oligandrum Po37 is used as a biocontrol agent of plant diseases. Here, we present the first draft of the P. oligandrum Po37 genome sequence, which comprises 725 scaffolds with a total length of 35.9 Mb and 11,695 predicted protein-coding genes. PMID:27081125

  12. Potential of a new strain of Bacillus amyloliquefaciens BUZ-14 as a biocontrol agent of postharvest fruit diseases.

    PubMed

    Calvo, H; Marco, P; Blanco, D; Oria, R; Venturini, M E

    2017-05-01

    The biocontrol potential of the Bacillus amyloliquefaciens strain BUZ-14 was tested against the main postharvest diseases of orange, apple, grape and stone fruit. After characterizing the temperature and pH growth curves of strain BUZ-14, its in vitro antifungal activity was determined against Botrytis cinerea, Monilinia fructicola, M. laxa, Penicillium digitatum, P. expansum and P. italicum. Subsequently, in vivo activity was tested against these pathogens by treating fruit with cells, endospores and cell-free supernatants. The in vitro results showed that BUZ-14 inhibited the growth of all the pathogens tested corresponding to the least susceptible species, P. italicum, and the most susceptible, M. laxa. In vivo tests corroborated these results as most of the treatments decreased the incidence of brown rot in stone fruit from 100% to 0%, establishing 10(7) CFU mL(-1) as the minimum inhibitory concentration. For the Penicillium species a preventive treatment inhibited P. digitatum and P. italicum growth in oranges and reduced P. expansum incidence in apples from 100% to 20%. Finally, it has been demonstrated that BUZ-14 was able to survive and to control brown rot in peaches stored at cool temperatures, making it a very suitable biocontrol agent for application during the post-harvest storage and marketing of horticultural products.

  13. Quorum-sensing signaling is required for production of the antibiotic pyrrolnitrin in a rhizospheric biocontrol strain of Serratia plymuthica.

    PubMed

    Liu, Xiaoguang; Bimerew, Mohammed; Ma, Yingxin; Müller, Henry; Ovadis, Marianna; Eberl, Leo; Berg, Gabriele; Chernin, Leonid

    2007-05-01

    One mechanism that bacteria have adopted to regulate the production of antimicrobial compounds is population-density-dependent LuxRI-type quorum sensing (QS), exploiting the production of N-acyl homoserine lactone (AHL) autoinducer signals. In biocontrol bacteria, most known cases involve the AHL control of phenazine antibiotics production by rhizospheric pseudomonads. This work is the first to demonstrate that phenazines are not the only group of biocontrol-related antibiotics whose production is regulated by QS systems. Strain HRO-C48 of Serratia plymuthica isolated from the rhizosphere of oilseed rape and described as a chitinolytic bacterium, which protects crops against Verticillium wilt, was also shown to produce wide-range antibiotic pyrrolnitrin and several AHLs, including N-butanoyl-HSL, N-hexanoyl-HSL and N-3-oxo-hexanoyl-HSL (OHHL). The genes splI and splR, which are analogues of luxI and luxR genes from other Gram-negative bacteria, were cloned and sequenced. The mutant AHL-4 (splI::miniTn5) was simultaneously deficient in the production of AHLs and pyrrolnitrin, as well as in its ability to suppress the growth of several fungal plant pathogens in vitro. However, pyrrolnitrin production could be restored in this mutant by introduction of the splIR genes cloned into a plasmid or by addition of the conditioned medium from strain C48 or OHHL standard to the growth medium.

  14. Complete Genome Sequence of the Biocontrol Strain Pseudomonas protegens Cab57 Discovered in Japan Reveals Strain-Specific Diversity of This Species

    PubMed Central

    Takeuchi, Kasumi; Noda, Naomi; Someya, Nobutaka

    2014-01-01

    The biocontrol strain Pseudomonas sp. Cab57 was isolated from the rhizosphere of shepherd’s purse growing in a field in Hokkaido by screening the antibiotic producers. The whole genome sequence of this strain was obtained by paired-end and whole-genome shotgun sequencing, and the gaps between the contigs were closed using gap-spanning PCR products. The P. sp. Cab57 genome is organized into a single circular chromosome with 6,827,892 bp, 63.3% G+C content, and 6,186 predicted protein-coding sequences. Based on 16S rRNA gene analysis and whole genome analysis, strain Cab57 was identified as P. protegens. As reported in P. protegens CHA0 and Pf-5, four gene clusters (phl, prn, plt, and hcn) encoding the typical antibiotic metabolites and the reported genes associated with Gac/Rsm signal transduction pathway of these strains are fully conserved in the Cab57 genome. Actually strain Cab57 exhibited typical Gac/Rsm activities and antibiotic production, and these activities were enhanced by knocking out the retS gene (for a sensor kinase acting as an antagonist of GacS). Two large segments (79 and 115 kb) lacking in the Cab57 genome, as compared with the Pf-5 genome, accounted for the majority of the difference (247 kb) between these genomes. One of these segments was the complete rhizoxin analog biosynthesis gene cluster (ca. 79 kb) and another one was the 115-kb mobile genomic island. A whole genome comparison of those relative strains revealed that each strain has unique gene clusters involved in metabolism such as nitrite/nitrate assimilation, which was identified in the Cab57 genome. These findings suggest that P. protegens is a ubiquitous bacterium that controls its biocontrol traits while building up strain-specific genomic repertoires for the biosynthesis of secondary metabolites and niche adaptation. PMID:24695768

  15. Identification of genetic defects in the atoxigenic biocontrol strain Aspergillus flavus K49 reveals the presence of a competitive recombinant group in field populations

    USDA-ARS?s Scientific Manuscript database

    Contamination of corn, cotton, peanuts and tree nuts by aflatoxins is a severe economic burden for growers. A current biocontrol strategy is to use non-aflatoxigenic Aspergillus flavus strains to competitively exclude field toxigenic Aspergillus species. Aspergillus flavus K49 does not produce aflat...

  16. Draft Genome Sequences of Pseudomonas fluorescens Strains SF39a and SF4c, Potential Plant Growth Promotion and Biocontrol Agents

    PubMed Central

    Ly, Lindsey K.; Underwood, Grace E.; McCully, Lucy M.; Bitzer, Adam S.; Godino, Agustina; Bucci, Vanni; Brigham, Christopher J.; Príncipe, Analía; Fischer, Sonia E.

    2015-01-01

    Pseudomonas fluorescens SF4c and SF39a, strains isolated from wheat rhizosphere, have potential applications in plant growth promotion and biocontrol of fungal diseases of crop plants. We report the draft genome sequences of SF4c and SF39a with estimated sizes of 6.5 Mb and 5.9 Mb, respectively. PMID:25814613

  17. Statistical test for tolerability of effects of an antifungal biocontrol strain on fungal communities in three arable soils.

    PubMed

    Antweiler, Kai; Schreiter, Susanne; Keilwagen, Jens; Baldrian, Petr; Kropf, Siegfried; Smalla, Kornelia; Grosch, Rita; Heuer, Holger

    2017-03-01

    A statistical method was developed to test for equivalence of microbial communities analysed by next-generation sequencing of amplicons. The test uses Bray-Curtis distances between the microbial community structures and is based on a two-sample jackknife procedure. This approach was applied to investigate putative effects of the antifungal biocontrol strain RU47 on fungal communities in three arable soils which were analysed by high-throughput ITS amplicon sequencing. Two contrasting workflows to produce abundance tables of operational taxonomic units from sequence data were applied. For both, the developed test indicated highly significant equivalence of the fungal communities with or without previous exposure to RU47 for all soil types, with reference to fungal community differences in conjunction with field site or cropping history. However, minor effects of RU47 on fungal communities were statistically significant using highly sensitive multivariate tests. Nearly all fungal taxa responding to RU47 increased in relative abundance indicating the absence of ecotoxicological effects. Use of the developed equivalence test is not restricted to evaluate effects on soil microbial communities by inoculants for biocontrol, bioremediation or other purposes, but could also be applied for biosafety assessment of compounds like pesticides, or genetically engineered plants. © 2017 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  18. Colonization Pattern of the Biocontrol Strain Pseudomonas chlororaphis MA 342 on Barley Seeds Visualized by Using Green Fluorescent Protein

    PubMed Central

    Tombolini, Riccardo; van der Gaag, Dirk Jan; Gerhardson, Berndt; Jansson, Janet K.

    1999-01-01

    Pseudomonas chlororaphis MA 342 is a potent biocontrol agent that can be used against several seed-borne diseases of cereal crops, including net blotch of barley caused by the fungus Drechslera teres. In this study, strain MA 342 was tagged with the gfp gene (encoding the green fluorescent protein) in order to study the fate of cells after seed inoculation. The gfp-tagged strain, MA 342G2, had the same biocontrol efficacy as the wild type when it was applied at high cell concentrations to seeds but was less effective at lower cell concentrations. By comparing cell counts determined by microscopy to the number of CFU, we found that the number of culturable cells was significantly lower than the total number of bacteria on seeds which were inoculated and dried for 20 h. Confocal microscopy and epifluorescence stereomicroscopy were used to determine the pattern of MA 342G2 colonization and cell aggregation on barley seeds. Immediately after inoculation of seeds, bacteria were found mainly under the seed glume, and there was no particular aggregation pattern. However, after the seeds were sown, irregularly distributed areas of bacterial aggregation were found, which reflected epiphytic colonization of glume cells. There was a trend towards bacterial aggregation near the embryo but never within the embryo. Bacterial aggregates were regularly found in the groove of each seed formed by the base of the coleoptile and the scutellum. Based on these results, we suggest that MA 342 colocalizes with the pathogen D. teres, which facilitates the action of the fungistatic compound(s) produced by this strain. PMID:10427065

  19. Effects of culture conditions on spore types of Clonostachys rosea 67-1 in submerged fermentation.

    PubMed

    Sun, M H; Chen, Y M; Liu, J F; Li, S D; Ma, G Z

    2014-04-01

    The promising biocontrol isolate Clonostachys rosea 67-1 was investigated to clarify the effects of culture conditions on chlamydospore production in submerged fermentation. Culture conditions significantly affected both performance and types of C. rosea sporulation. C. rosea 67-1 was hard to generate chlamydospores under conventional conditions. However, the proportion of resistant spores increased to 17·4 and 15·5% in PD and rice meal media, respectively, in 8 days. Chlamydospore productivity was boosted (>threefold) with the addition of 50-200 mg l(-1) CuSO4 . The pH of the medium played a vital role in 67-1 sporulation. The percentage of chlamydospores decreased rapidly with increased pH (88·1% at pH 3·0 to 1·0% at pH 6·5). The optimal pH for conidia production was 6·0-6·5, at which chlamydospore forming was strongly inhibited. Regulating pH during fermentation contributed to improving output and proportion of resistant spores. When 67-1 was inoculated into broth with an initial pH of 6·5, followed by adjustment to pH 3·5 after 48 h, the number of chlamydospores reached 1·1 × 10(8) ml(-1). The impact of temperature and rotational speed was also analysed; an ultimate capacity of chlamydospores was achieved at 30°C and the speed above 120 rev min(-1) (P < 0·05). Clonostachys rosea is one of the most promising biocontrol agents in countering many plant fungal diseases. However, large-scale production and commercialization are hampered by the lack of understanding of the impacts of culture conditions on performance and types of C. rosea sporulation and subsequently inadequate research on the techniques for chlamydospore production. In addressing these concerns, this study provides a unique insight into the manipulation of C. rosea sporulation and chlamydospore fermentation of the biocontrol fungus. © 2013 The Society for Applied Microbiology.

  20. Response of tomato wilt pathogen Ralstonia solanacearum to the volatile organic compounds produced by a biocontrol strain Bacillus amyloliquefaciens SQR-9

    PubMed Central

    Raza, Waseem; Ling, Ning; Yang, Liudong; Huang, Qiwei; Shen, Qirong

    2016-01-01

    It is important to study the response of plant pathogens to the antibiosis traits of biocontrol microbes to design the efficient biocontrol strategies. In this study, we evaluated the role of volatile organic compounds (VOCs) produced by a biocontrol strain Bacillus amyloliquefaciens SQR-9 on the growth and virulence traits of tomato wilt pathogen Ralstonia solanacearum (RS). The VOCs of SQR-9 significantly inhibited the growth of RS on agar medium and in soil. In addition, the VOCs significantly inhibited the motility traits, production of antioxidant enzymes and exopolysaccharides, biofilm formation and tomato root colonization by RS. The strain SQR-9 produced 22 VOCs, but only nine VOCs showed 1–11% antibacterial activity against RS in their corresponding amounts; however, the consortium of all VOCs showed 70% growth inhibition of RS. The proteomics analysis showed that the VOCs of SQR-9 downregulated RS proteins related to the antioxidant activity, virulence, carbohydrate and amino acid metabolism, protein folding and translation, while the proteins involved in the ABC transporter system, amino acid synthesis, detoxification of aldehydes and ketones, methylation, protein translation and folding, and energy transfer were upregulated. This study describes the significance and effectiveness of VOCs produced by a biocontrol strain against tomato wilt pathogen. PMID:27103342

  1. Response of tomato wilt pathogen Ralstonia solanacearum to the volatile organic compounds produced by a biocontrol strain Bacillus amyloliquefaciens SQR-9.

    PubMed

    Raza, Waseem; Ling, Ning; Yang, Liudong; Huang, Qiwei; Shen, Qirong

    2016-04-22

    It is important to study the response of plant pathogens to the antibiosis traits of biocontrol microbes to design the efficient biocontrol strategies. In this study, we evaluated the role of volatile organic compounds (VOCs) produced by a biocontrol strain Bacillus amyloliquefaciens SQR-9 on the growth and virulence traits of tomato wilt pathogen Ralstonia solanacearum (RS). The VOCs of SQR-9 significantly inhibited the growth of RS on agar medium and in soil. In addition, the VOCs significantly inhibited the motility traits, production of antioxidant enzymes and exopolysaccharides, biofilm formation and tomato root colonization by RS. The strain SQR-9 produced 22 VOCs, but only nine VOCs showed 1-11% antibacterial activity against RS in their corresponding amounts; however, the consortium of all VOCs showed 70% growth inhibition of RS. The proteomics analysis showed that the VOCs of SQR-9 downregulated RS proteins related to the antioxidant activity, virulence, carbohydrate and amino acid metabolism, protein folding and translation, while the proteins involved in the ABC transporter system, amino acid synthesis, detoxification of aldehydes and ketones, methylation, protein translation and folding, and energy transfer were upregulated. This study describes the significance and effectiveness of VOCs produced by a biocontrol strain against tomato wilt pathogen.

  2. Phenazine-1-carboxylic acid is a more important contributor to biocontrol Fusarium oxysporum than pyrrolnitrin in Pseudomonas fluorescens strain Psd.

    PubMed

    Upadhyay, Ashutosh; Srivastava, Sheela

    2011-05-20

    Phenazines and pyrrolnitrin (Prn) are broad spectrum antibiotics, produced by bacteria, more so by the biocontrol strains to kill the phytopathogens in soil. We have studied a rhizospheric soil isolate of Pseudomonas fluorescens strain Psd producing both phenazine-1-carboxylic acid (PCA) and Prn. In order to study the contribution of these antibiotics, the phzD and prnC genes involved in PCA and Prn biosynthesis, were disrupted in a site-specific manner using a group II intron-based Targetron gene-knockout system, and gene disruption followed by allelic exchange through homologous recombination, respectively. The resulting knockout strains Psdphz122s-34 and PsdprnC::gen did not produce PCA and Prn, respectively. In fact, by combining these two strategies, a Psdphz122s-34prnC::gen double mutant could also be generated. Identification and lack of PCA production was corroborated by HPLC/APCI-MS analysis, and TLC detection for both the antibiotics in these mutants. Loss of antifungal activity against the phytopathogenic fungus Fusarium oxysporum was observed using in vitro growth assays on plates or growth chamber experiments with tomato seedling on an artificial substrate. Based on the characterization of these gene knockout mutants, we propose that PCA and Prn have a major role in antifungal activity of strain Psd. Copyright © 2010 Elsevier GmbH. All rights reserved.

  3. Studies of plant colonisation by closely related Bacillus amyloliquefaciens biocontrol agents using strain specific quantitative PCR assays.

    PubMed

    Johansson, Anna H; Bejai, Sarosh; Niazi, Adnan; Manzoor, Shahid; Bongcam-Rudloff, Erik; Meijer, Johan

    2014-12-01

    Certain strains of Bacillus amyloliquefaciens can colonize plants and improve growth and stress management. In order to study these effects, bacterial growth dynamics on plants and in the rhizosphere are of interest calling for specific analytical tools. For that purpose, quantitative real-time PCR (qPCR) assays were developed in order to differentiate among three closely related B. amyloliquefaciens subsp. plantarum strains (UCMB5033, UCMB5036, UCMB5113) and to determine their levels with high accuracy. Oligonucleotide primers were designed for strain unique gene sequences and used for SYBR green based qPCR analysis. Standard curves covered a wide linear range (10(6)) of DNA amounts with the lowest detection level at 50 fg. Post-reaction melting curve analysis showed only a single product. Accurate threshold cycles were obtained, even in the presence of high excess of related Bacillus strains and total bacterial DNA from soil. Analysis of Bacillus colonisation after seed treatment of two oilseed rape cultivars (Oase and Ritz) grown on agar support showed a time dependent effect but that the bacteria mostly were found on root tissues and little on green tissues. The colonisation on plants grown in soil varied among the Bacillus strains where Oase seemed to house more bacteria than Ritz. Applied as a mixture, all three Bacillus strains co-existed on the roots of plants grown in soil. The qPCR assay in combination with other techniques will be a powerful tool to study plant interactions of these B. amyloliquefaciens biocontrol agents to further understand the requirements for successful interactions and improvement of plant properties.

  4. Formulation development of the biocontrol agent Bacillus subtilis strain CPA-8 by spray-drying.

    PubMed

    Yánez-Mendizábal, V; Viñas, I; Usall, J; Torres, R; Solsona, C; Abadias, M; Teixidó, N

    2012-05-01

    To prepare commercially acceptable formulations of Bacillus subtilis CPA-8 by spray-drying with long storage life and retained efficacy to control peach and nectarine brown rot caused by Monilinia spp. CPA-8 24-h- and 72-h-old cultures were spray dried using 10% skimmed milk, 10% skimmed milk plus 10% MgSO(4) , 10% MgSO(4) and 20% MgSO(4) as carriers/protectants. All carriers/protectants gave good percentages of powder recovery (28-38%) and moisture content (7-13%). CPA-8 survival varied considerably among spray-dried 24-h- and 72-h-old cultures. Seventy-two hours culture spray dried formulations showed the highest survival (28-32%) with final concentration products of 1·6-3·3 × 10(9) CFU g(-1) , while viability of 24-h-old formulations was lower than 1%. Spray-dried 72-h-old formulations were selected to subsequent evaluation. Rehydration of cells with water provided a good recovery of CPA-8 dried cells, similar to other complex rehydration media tested. Spray-dried formulations stored at 4 ± 1 and 20 ± 1°C showed good shelf life during 6 months, and viability was maintained or slightly decreased by 0·2-0·3-log. CPA-8 formulations after 4- and 6 months storage were effective in controlling brown rot caused by Monilinia spp. on nectarines and peaches resulting in a 90-100% reduction in disease incidence. Stable and effective formulations of biocontrol agent B. subtilis CPA-8 could be obtained by spray-drying. New shelf-stable and effective formulations of a biocontrol agent have been obtained by spray-drying to control brown rot on peach. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  5. Influence of host plant genotype, presence of a pathogen, and coinoculation with Pseudomonas fluorescens strains on the rhizosphere expression of hydrogen cyanide- and 2,4-diacetylphloroglucinol biosynthetic genes in P. fluorescens biocontrol strain CHA0.

    PubMed

    Jamali, Fatemeh; Sharifi-Tehrani, Abbas; Lutz, Matthias P; Maurhofer, Monika

    2009-02-01

    The production of hydrogen cyanide (HCN) and 2,4-diacetylphloroglucinol (DAPG) is a major factor in the control of soil-borne diseases by Pseudomonas fluorescens CHA0. We investigated the impact of different biotic factors on the expression of HCN-in comparison to DAPG biosynthetic genes in the rhizosphere. To this end, the influence of plant cultivar, pathogen infection, and coinoculation with other biocontrol strains on the expression of hcnA-lacZ and phlA-lacZ fusion in strain CHA0 was monitored on the roots of bean. Interestingly, all the tested factors influenced the expression of the two biocontrol traits in a similar way. For both genes, we observed a several-fold higher expression in the rhizosphere of cv. Derakhshan compared with cvs. Goli and Naz, although bacterial rhizosphere colonization levels were similar on all cultivars tested. Root infection by Rhizoctonia solani stimulated total phlA and hcnA gene expression in the bean rhizosphere. Coinoculation of strain CHA0 with DAPG-producing P. fluorescens biocontrol strains Pf-68 and Pf-100 did neither result in a substantial alteration of hcnA nor of phlA expression in CHA0 on bean roots. To our best knowledge, this is the first study investigating the impact of biotic factors on HCN production by a bacterial biocontrol strain in the rhizosphere.

  6. Putative use of a Bacillus subtilis L194 strain for biocontrol of Phoma medicaginis in Medicago truncatula seedlings.

    PubMed

    Ben Slimene, Imen; Tabbene, Olfa; Djebali, Naceur; Cosette, Pascal; Schmitter, Jean Marie; Jouenne, Thierry; Urdaci, Maria-Camino; Limam, Ferid

    2012-06-01

    An antagonist L194 strain against Phoma medicaginis pathogenic fungi was isolated from Tunisian soil (vicinity of Tunis) and identified as Bacillus subtilis based on biochemical characteristics and partial 16S rDNA sequence. When cells were grown in a minimal medium for 24 h, spore culture supernatant exhibited 2-fold higher antifungal activity than vegetative cells. MALDI-TOF mass spectrometry analysis showed that L194 spores produced mainly iturins, surfactins and fengycins with long-chain fatty acids, and other not yet identified compounds. Both vegetative cells and spores of L194 efficiently reduced germination of P. medicaginis conidia. As revealed by atomic force microscopy, L194 spores modified conidia morphology from a regular to a deflated shape. Data suggest that lipopeptides interacted with the cytoplasmic membrane, causing pore formation. In vivo, L194 spores were highly protective against P. medicaginis by reducing disease symptoms and alleviating growth disturbance of Medicago truncatula seedlings. As a whole, the lipopeptide-producing L194 strain may be successfully used in biocontrol of plant diseases induced by phytopathogenic fungi such as P. medicaginis.

  7. Complete genome sequence of Bacillus velezensis LM2303, a biocontrol strain isolated from the dung of wild yak inhabited Qinghai-Tibet plateau.

    PubMed

    Chen, Liang

    2017-06-10

    Bacillus velezensis LM2303 is a biocontrol strain with a broad inhibitory spectrum against plant pathogens, isolated from the dung of wild yak inhabited Qinghai-Tibet plateau, China. Here we present its complete genome sequence, which consists of a single, circular chromosome of 3,989,393bp with a 46.68% G+C content. Genome analysis revealed genes encoding specialized functions for the biosynthesis of antifungal metabolites and antibacterial metabolites, the promotion of plant growth, the alleviation of oxidative stress and nutrient utilization. And the biosynthesis of antimicrobial metabolites in strain LM2303 was confirmed by biochemical analysis, while its plant growth promoting traits were confirmed by inoculation tests. Our results will establish a better foundation for further studies and biocontrol application of B. velezensis LM2303. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Complete genome sequence of Pseudomonas fluorescens strain PICF7, an indigenous root endophyte from olive (Olea europaea L.) and effective biocontrol agent against Verticillium dahliae

    PubMed Central

    2015-01-01

    Pseudomonas fluorescens strain PICF7 is a native endophyte of olive roots. Previous studies have shown this motile, Gram-negative, non-sporulating bacterium is an effective biocontrol agent against the soil-borne fungus Verticillium dahliae, the causal agent of one of the most devastating diseases for olive (Olea europaea L.) cultivation. Here, we announce and describe the complete genome sequence of Pseudomonas fluorescens strain PICF7 consisting of a circular chromosome of 6,136,735 bp that encodes 5,567 protein-coding genes and 88 RNA-only encoding genes. Genome analysis revealed genes predicting factors such as secretion systems, siderophores, detoxifying compounds or volatile components. Further analysis of the genome sequence of PICF7 will help in gaining insights into biocontrol and endophytism. PMID:25685259

  9. Draft Genome Sequences of Pseudomonas fluorescens Strains SF39a and SF4c, Potential Plant Growth Promotion and Biocontrol Agents.

    PubMed

    Ly, Lindsey K; Underwood, Grace E; McCully, Lucy M; Bitzer, Adam S; Godino, Agustina; Bucci, Vanni; Brigham, Christopher J; Príncipe, Analía; Fischer, Sonia E; Silby, Mark W

    2015-03-26

    Pseudomonas fluorescens SF4c and SF39a, strains isolated from wheat rhizosphere, have potential applications in plant growth promotion and biocontrol of fungal diseases of crop plants. We report the draft genome sequences of SF4c and SF39a with estimated sizes of 6.5 Mb and 5.9 Mb, respectively. Copyright © 2015 Ly et al.

  10. Draft Genome Sequence of Bacillus velezensis 3A-25B, a Strain with Biocontrol Activity against Fungal and Oomycete Root Plant Phytopathogens, Isolated from Grassland Soil.

    PubMed

    Martínez-Raudales, Inés; De La Cruz-Rodríguez, Yumiko; Vega-Arreguín, Julio; Alvarado-Gutiérrez, Alejandro; Fraire-Mayorga, Atzin; Alvarado-Rodríguez, Miguel; Balderas-Hernández, Victor; Gómez-Soto, José Manuel; Fraire-Velázquez, Saúl

    2017-09-28

    Here, we present the draft genome of Bacillus velezensis 3A-25B, which totaled 4.01 Mb with 36 contigs, 3,948 genes, and a GC content of 46.34%. This strain, which demonstrates biocontrol activity against root rot causal phytopathogens in horticultural crops and friendly interactions in roots of pepper plantlets, was obtained from grassland soil in Zacatecas Province, Mexico. Copyright © 2017 Martínez-Raudales et al.

  11. Draft Genome Sequence of Bacillus velezensis 3A-25B, a Strain with Biocontrol Activity against Fungal and Oomycete Root Plant Phytopathogens, Isolated from Grassland Soil

    PubMed Central

    Martínez-Raudales, Inés; De La Cruz-Rodríguez, Yumiko; Vega-Arreguín, Julio; Alvarado-Gutiérrez, Alejandro; Fraire-Mayorga, Atzin; Alvarado-Rodríguez, Miguel; Balderas-Hernández, Victor; Gómez-Soto, José Manuel

    2017-01-01

    ABSTRACT Here, we present the draft genome of Bacillus velezensis 3A-25B, which totaled 4.01 Mb with 36 contigs, 3,948 genes, and a GC content of 46.34%. This strain, which demonstrates biocontrol activity against root rot causal phytopathogens in horticultural crops and friendly interactions in roots of pepper plantlets, was obtained from grassland soil in Zacatecas Province, Mexico. PMID:28963212

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

    PubMed

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

    2015-05-01

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

  13. High-Quality Draft Genome Sequence of Biocontrol Strain Pantoea sp. OXWO6B1.

    PubMed

    Town, Jennifer; Audy, Patrice; Boyetchko, Susan M; Dumonceaux, Tim J

    2016-06-23

    Pantoea sp. strain OXWO6B1 inhibits the growth of the potato pathogen Phytophthora infestans We determined the 5.2-Mbp genome sequence of this strain, which featured at least 3 confirmed plasmids of up to 250 kbp. The genome sequence of OXWO6B1 is different from that of all previously sequenced strains of Pantoea. Copyright © 2016 Town et al.

  14. Novel Trichoderma polysporum Strain for the Biocontrol of Pseudogymnoascus destructans, the Fungal Etiologic Agent of Bat White Nose Syndrome.

    PubMed

    Zhang, Tao; Chaturvedi, Vishnu; Chaturvedi, Sudha

    2015-01-01

    White-nose syndrome (WNS), an emerging disease of hibernating bats, has rapidly spread across eastern North America killing millions of bats. Pseudogymnoascus destructans (Pd), the sole etiologic agent of WNS, is widespread and persistent in bat hibernacula. Control of Pd in the affected sites is urgently needed to break the transmission cycle while minimizing any adverse impact on the native organisms. We isolated a novel strain of Trichoderma polysporum (Tp) from one of the caves at the epicenter of WNS zoonotic. Detailed experimental studies revealed: (1) Tp WPM 39143 was highly adapted to grow at temperatures simulating the cave environment (6°C-15°C), (2) Tp WPM 39143 restricted Pd colony growth in dual culture challenges, (3) Tp WPM 39143 caused four logs reduction of Pd colony forming units and genome copies in autoclaved soil samples from one of the WNS affected caves, (4) Tp WPM 39143 extract showed specific fungicidal activity against Pd in disk diffusion assay, but not against closely related fungus P. pannorum (Pp), (5) Tp WPM 39143 extract retained inhibitory activity after exposure to high temperatures, light and proteinase K, and (6) Inhibitory metabolites in Tp WPM 39143 extract comprised of water-soluble, high polarity compounds. These results suggest that Tp WPM 39143 is a promising candidate for further evaluation as a biocontrol agent of Pd in WNS affected sites.

  15. Production of trichodiene by Trichoderma harzianum alters the perception of this biocontrol strain by plants and antagonized fungi.

    PubMed

    Malmierca, Mónica G; McCormick, Susan P; Cardoza, Rosa E; Alexander, Nancy J; Monte, Enrique; Gutiérrez, Santiago

    2015-08-01

    Trichothecenes are phytotoxic sesquiterpenic mycotoxins that can act as virulence factors in plant diseases. Harzianum A (HA) is a non-phytotoxic trichothecene produced by Trichoderma arundinaceum. The first step in HA biosynthesis is the conversion of farnesyl diphosphate to trichodiene (TD), a volatile organic compound (VOC), catalysed by a sesquiterpene synthase encoded by the tri5 gene. Expression of tri5 in the biocontrol strain Trichoderma harzianum CECT 2413 resulted in production of TD in parallel with a reduction of ergosterol biosynthesis and an unexpected increase in the level of squalene. Transformants expressing tri5 displayed low chitinase activity and induced expression of Botrytis cinerea BOT genes, although their total antagonistic potential against phytopathogenic fungi was not reduced. VOCs released by the tri5-transformant induced expression of tomato defence genes related to salicylic acid (SA), and TD itself strongly induced the expression of SA-responsive genes and reduced the development of lateral roots. Together, these results suggest that TD acts as a signalling VOC in the interactions of Trichoderma with plants and other microorganisms by modulating the perception of this fungus to a given environment. Moreover, the TD ability to induce systemic defences indicates that complex trichothecene structures may not be necessary for inducing such responses.

  16. Novel Trichoderma polysporum Strain for the Biocontrol of Pseudogymnoascus destructans, the Fungal Etiologic Agent of Bat White Nose Syndrome

    PubMed Central

    Zhang, Tao; Chaturvedi, Vishnu; Chaturvedi, Sudha

    2015-01-01

    White-nose syndrome (WNS), an emerging disease of hibernating bats, has rapidly spread across eastern North America killing millions of bats. Pseudogymnoascus destructans (Pd), the sole etiologic agent of WNS, is widespread and persistent in bat hibernacula. Control of Pd in the affected sites is urgently needed to break the transmission cycle while minimizing any adverse impact on the native organisms. We isolated a novel strain of Trichoderma polysporum (Tp) from one of the caves at the epicenter of WNS zoonotic. Detailed experimental studies revealed: (1) Tp WPM 39143 was highly adapted to grow at temperatures simulating the cave environment (6°C-15°C), (2) Tp WPM 39143 restricted Pd colony growth in dual culture challenges, (3) Tp WPM 39143 caused four logs reduction of Pd colony forming units and genome copies in autoclaved soil samples from one of the WNS affected caves, (4) Tp WPM 39143 extract showed specific fungicidal activity against Pd in disk diffusion assay, but not against closely related fungus P. pannorum (Pp), (5) Tp WPM 39143 extract retained inhibitory activity after exposure to high temperatures, light and proteinase K, and (6) Inhibitory metabolites in Tp WPM 39143 extract comprised of water-soluble, high polarity compounds. These results suggest that Tp WPM 39143 is a promising candidate for further evaluation as a biocontrol agent of Pd in WNS affected sites. PMID:26509269

  17. Functional genomic approaches for understanding the mode of action of Bacillus sp biocontrol strains

    USDA-ARS?s Scientific Manuscript database

    Complete genome sequencing of several Bacillus sp. strains has shed new light on the mode of action of these antagonists of plant pathogens. The use of genomic data mining tools provided the ability to quickly determine the potential of these strains to produce bioactive secondary metabolites. Our B...

  18. Genomic comparisons of two Bacillus subtilis biocontrol strains with different modes of actions

    USDA-ARS?s Scientific Manuscript database

    Bacillus subtilis strains AS 43.3 and OH131.1 were isolated from wheat anthers and shown to be efficacious in managing Fusarium head blight in greenhouse and some field trials. Chemical analysis of the cell-free culture supernatant identified B. subtilis strain AS 43.3 to be a potent producer of the...

  19. Characterization of the Biosynthetic Operon for the Antibacterial Peptide Herbicolin in Pantoea vagans Biocontrol Strain C9-1 and Incidence in Pantoea Species

    PubMed Central

    Kamber, Tim; Lansdell, Theresa A.; Stockwell, Virginia O.; Ishimaru, Carol A.; Smits, Theo H. M.

    2012-01-01

    Pantoea vagans C9-1 is a biocontrol strain that produces at least two antibiotics inhibiting the growth of Erwinia amylovora, the causal agent of fire blight disease of pear and apple. One antibiotic, herbicolin I, was purified from culture filtrates of P. vagans C9-1 and determined to be 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine, also known as Nß-epoxysuccinamoyl-DAP-valine. A plasposon library was screened for mutants that had lost the ability to produce herbicolin I. It was shown that mutants had reduced biocontrol efficacy in immature pear assays. The biosynthetic gene cluster in P. vagans C9-1 was identified by sequencing the flanking regions of the plasposon insertion sites. The herbicolin I biosynthetic gene cluster consists of 10 coding sequences (CDS) and is located on the 166-kb plasmid pPag2. Sequence comparisons identified orthologous gene clusters in Pantoea agglomerans CU0119 and Serratia proteamaculans 568. A low incidence of detection of the biosynthetic cluster in a collection of 45 Pantoea spp. from biocontrol, environmental, and clinical origins showed that this is a rare trait among the tested strains. PMID:22504810

  20. Characterization of the biosynthetic operon for the antibacterial peptide herbicolin in Pantoea vagans biocontrol strain C9-1 and incidence in Pantoea species.

    PubMed

    Kamber, Tim; Lansdell, Theresa A; Stockwell, Virginia O; Ishimaru, Carol A; Smits, Theo H M; Duffy, Brion

    2012-06-01

    Pantoea vagans C9-1 is a biocontrol strain that produces at least two antibiotics inhibiting the growth of Erwinia amylovora, the causal agent of fire blight disease of pear and apple. One antibiotic, herbicolin I, was purified from culture filtrates of P. vagans C9-1 and determined to be 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine, also known as N(ß)-epoxysuccinamoyl-DAP-valine. A plasposon library was screened for mutants that had lost the ability to produce herbicolin I. It was shown that mutants had reduced biocontrol efficacy in immature pear assays. The biosynthetic gene cluster in P. vagans C9-1 was identified by sequencing the flanking regions of the plasposon insertion sites. The herbicolin I biosynthetic gene cluster consists of 10 coding sequences (CDS) and is located on the 166-kb plasmid pPag2. Sequence comparisons identified orthologous gene clusters in Pantoea agglomerans CU0119 and Serratia proteamaculans 568. A low incidence of detection of the biosynthetic cluster in a collection of 45 Pantoea spp. from biocontrol, environmental, and clinical origins showed that this is a rare trait among the tested strains.

  1. Saprotrophic competitiveness and biocontrol fitness of a genetically modified strain of the plant-growth-promoting fungus Trichoderma hamatum GD12.

    PubMed

    Ryder, Lauren S; Harris, Beverley D; Soanes, Darren M; Kershaw, Michael J; Talbot, Nicholas J; Thornton, Christopher R

    2012-01-01

    Trichoderma species are ubiquitous soil fungi that hold enormous potential for the development of credible alternatives to agrochemicals and synthetic fertilizers in sustainable crop production. In this paper, we show that substantial improvements in plant productivity can be met by genetic modification of a plant-growth-promoting and biocontrol strain of Trichoderma hamatum, but that these improvements are obtained in the absence of disease pressure only. Using a quantitative monoclonal antibody-based ELISA, we show that an N-acetyl-β-d-glucosaminidase-deficient mutant of T. hamatum, generated by insertional mutagenesis of the corresponding gene, has impaired saprotrophic competitiveness during antagonistic interactions with Rhizoctonia solani in soil. Furthermore, its fitness as a biocontrol agent of the pre-emergence damping-off pathogen Sclerotinia sclerotiorum is significantly reduced, and its ability to promote plant growth is constrained by the presence of both pathogens. This work shows that while gains in T. hamatum-mediated plant-growth-promotion can be met through genetic manipulation of a single beneficial trait, such a modification has negative impacts on other aspects of its biology and ecology that contribute to its success as a saprotrophic competitor and antagonist of soil-borne pathogens. The work has important implications for fungal morphogenesis, demonstrating a clear link between hyphal architecture and secretory potential. Furthermore, it highlights the need for a holistic approach to the development of genetically modified Trichoderma strains for use as crop stimulants and biocontrol agents in plant agriculture.

  2. Persistence and cell culturability of biocontrol strain Pseudomonas fluorescens CHA0 under plough pan conditions in soil and influence of the anaerobic regulator gene anr.

    PubMed

    Mascher, Fabio; Schnider-Keel, Ursula; Haas, Dieter; Défago, Geneviève; Moënne-Loccoz, Yvan

    2003-02-01

    Certain fluorescent pseudomonads can protect plants from soil-borne pathogens, and it is important to understand how these biocontrol agents survive in soil. The persistence of the biocontrol strain Pseudomonas fluorescens CHA0-Rif under plough pan conditions was assessed in non-sterile soil microcosms by counting total cells (immunofluorescence microscopy), intact cells (BacLight membrane permeability test), viable cells (Kogure's substrate-responsiveness test) and culturable cells (colony counts on selective plates) of the inoculant. Viable but non-culturable cells of CHA0-Rif (106 cells g-1 soil) were found in flooded microcosms amended with fermentable organic matter, in which the soil redox potential was low (plough pan conditions), in agreement with previous observations of plough pan samples from a field inoculated with CHA0-Rif. However, viable but non-culturable cells were not found in unamended flooded, amended unflooded or unamended unflooded (i.e. control) microcosms, suggesting that such cells resulted from exposure of CHA0-Rif to a combination of low redox potential and oxygen limitation in soil. CHA0-Rif is strictly aerobic. Its anaerobic regulator ANR is activated by low oxygen concentrations and it controls production of the biocontrol metabolite hydrogen cyanide under microaerophilic conditions. Under plough pan conditions, an anr-deficient mutant of CHA0-Rif and its complemented derivative displayed the same persistence pattern as CHA0-Rif, indicating that anr was not implicated in the formation of viable but non-culturable cells of this strain at the plough pan.

  3. Identification of an antifungal metabolite produced by a potential biocontrol Actinomyces strain A01

    PubMed Central

    Lu, Cai Ge; Liu, Wei Cheng; Qiu, Ji Yan; Wang, Hui Min; Liu, Ting; De Liu, Wen

    2008-01-01

    Actinomyces strain A01 was isolated from soil of a vegetable field in the suburb of Beijing, China. According to the morphological, cultural, physiological and biochemical characteristics, and 16S rDNA sequence analysis, strain A01 was identified as Streptomyces lydicus. In the antimicrobial spectrum test strain A01 presented a stable and strong inhibitory activity against several plant pathogenic fungi such as Fusarium oxysporum, Botrytis cinerea, Monilinia laxa, etc. However, no antibacterial activity was found. In pot experiments in greenhouse, the development of tomato gray mold was markedly suppressed by treatment with the fermentation broth of the strain A01, and the control efficacy was higher than those of Pyrimethanil and Polyoxin. A main antifungal compound (purity 99.503%) was obtained from the fermentation broth of strain A01 using column chromatography and HPLC. The chemical structural analysis with U V, IR, MS, and NMR confirmed that the compound produced by the strain A01 is natamycin, a polyene antibiotic produced by S. chattanovgensis, S. natalensis, and S. gilvosporeus, widely used as a natural biological preservative for food according to previous reports. The present study revealed a new producing strain of natamycin and its potential application as a biological control agent for fungal plant diseases. PMID:24031293

  4. Analysis of Clonostachys rosea-induced resistance to tomato gray mold disease in tomato leaves.

    PubMed

    Mouekouba, Liana Dalcantara Ongouya; Zhang, Lili; Guan, Xin; Chen, Xiuling; Chen, Hongyu; Zhang, Jian; Zhang, Junfeng; Li, Jingfu; Yang, Yijun; Wang, Aoxue

    2014-01-01

    Tomato gray mold disease, caused by Botrytis cinerea, is a serious disease in tomato. Clonostachys rosea is an antagonistic microorganism to B. cinerea. To investigate the induced resistance mechanism of C. rosea, we examined the effects of these microorganisms on tomato leaves, along with changes in the activities of three defense enzymes (PAL, PPO, GST), second messengers (NO, H2O2, O2(-)) and phytohormones (IAA, ABA, GA3, ZT, MeJA, SA and C2H4). Compared to the control, all treatments induced higher levels of PAL, PPO and GST activity in tomato leaves and increased NO, SA and GA3 levels. The expression of WRKY and MAPK, two important transcription factors in plant disease resistance, was upregulated in C. rosea- and C. rosea plus B. cinerea-treated samples. Two-dimensional gel electrophoresis analysis showed that two abundant proteins were present in the C. rosea plus B. cinerea-treated samples but not in the other samples. These proteins were determined (by mass spectrum analysis) to be LEXYL2 (β-xylosidase) and ATP synthase CF1 alpha subunit. Therefore, C. rosea plus B. cinerea treatment induces gray mold resistance in tomato. This study provides a basis for elucidating the mechanism of C. rosea as a biocontrol agent.

  5. Efficacy of Clonostachys rosea and Duddingtonia flagrans in Reducing the Haemonchus contortus Infective Larvae

    PubMed Central

    da Silva, Manoel Eduardo; Braga, Fabio Ribeiro; de Gives, Pedro Mendoza; Uriostegui, Miguel Angel Mercado; Reyes, Manuela; Soares, Filippe Elias de Freitas; de Carvalho, Lorendane Millena; Rodrigues, Francielle Bosi; de Araújo, Jackson Victor

    2015-01-01

    The biocontrol is proven effective in reducing in vitro and in situ free-living stages of major gastrointestinal helminths, allowing progress in reducing losses by parasitism, maximizing production, and productivity. This study aimed at evaluating the predatory activity of fungal isolates of Duddingtonia flagrans and Clonostachys rosea species and its association on infective larvae (L3) of H. contortus in microplots formed by grasses and maintained in a protected environment. All groups were added with 10 mL of an aqueous suspension with 618 H. contortus L3 approximately. Group 1 was used as control and only received the infective larvae. Groups 2 and 3 received D. flagrans chlamydospores and C. rosea conidia at doses of 5 × 106. Group 4 received the combination of 5 × 106 D. flagrans chlamydospores + 5 × 106 C. rosea conidia. D. flagrans and C. rosea showed nematicidal effectiveness reducing by 91.5 and 88.9%, respectively, the population of H. contortus L3. However, when used in combination efficiency decreased to 74.5% predation of H. contortus L3. These results demonstrate the need for further studies to determine the existence of additive effects, synergistic or antagonistic, between these species. PMID:26504809

  6. Synergistic Effect of Dazomet Soil Fumigation and Clonostachys rosea Against Cucumber Fusarium Wilt.

    PubMed

    Tian, Tian; Li, Shi-Dong; Sun, Man-Hong

    2014-12-01

    Soil fumigation and biological control are two control measures frequently used against soilborne diseases. In this study, the chemical fumigant dazomet was applied in combination with the biocontrol agent (BCA) Clonostachys rosea 67-1 to combat cucumber wilt caused by Fusarium oxysporum f. sp. cucumerinum KW2-1. When the mycoparasite C. rosea 67-1 was applied after dazomet fumigation, disease control reached 100%, compared with 88.1 and 69.8% for dazomet and 67-1 agent, respectively, applied alone, indicating a synergistic effect of dazomet and C. rosea in combating cucumber Fusarium wilt based on analysis of Bliss Independence. To understand the synergistic mechanism, the effects of chemical fumigation on the colonization potential and activity of F. oxysporum f. sp. cucumerinum, and the interaction between the BCA and the pathogen were investigated. The results showed that growth of the pathogen decreased with increasing dazomet concentration subsequent to fumigation. When exposed to dazomet at 100 ppm, the fungal sporulation rate decreased by 94.4%. Severe damage was observed in fumigated isolates using scanning electron microscopy. In the greenhouse, disease incidence of cucumber caused by fumigated F. oxysporum f. sp. cucumerinum significantly decreased. Whereas germination of C. rosea 67-1 spores increased by >sixfold in fumigated soil, and its ability to parasitize fumigated F. oxysporum f. sp. cucumerinum significantly increased (P = 0.014).

  7. Analysis of Clonostachys rosea-Induced Resistance to Tomato Gray Mold Disease in Tomato Leaves

    PubMed Central

    Guan, Xin; Chen, Xiuling; Chen, Hongyu; Zhang, Jian; Zhang, Junfeng; Li, Jingfu; Yang, Yijun; Wang, Aoxue

    2014-01-01

    Tomato gray mold disease, caused by Botrytis cinerea, is a serious disease in tomato. Clonostachys rosea is an antagonistic microorganism to B. cinerea. To investigate the induced resistance mechanism of C. rosea, we examined the effects of these microorganisms on tomato leaves, along with changes in the activities of three defense enzymes (PAL, PPO, GST), second messengers (NO, H2O2, O2−) and phytohormones (IAA, ABA, GA3, ZT, MeJA, SA and C2H4). Compared to the control, all treatments induced higher levels of PAL, PPO and GST activity in tomato leaves and increased NO, SA and GA3 levels. The expression of WRKY and MAPK, two important transcription factors in plant disease resistance, was upregulated in C. rosea- and C. rosea plus B. cinerea-treated samples. Two-dimensional gel electrophoresis analysis showed that two abundant proteins were present in the C. rosea plus B. cinerea-treated samples but not in the other samples. These proteins were determined (by mass spectrum analysis) to be LEXYL2 (β-xylosidase) and ATP synthase CF1 alpha subunit. Therefore, C. rosea plus B. cinerea treatment induces gray mold resistance in tomato. This study provides a basis for elucidating the mechanism of C. rosea as a biocontrol agent. PMID:25061981

  8. Evaluation of Atoxigenic Strains of Aspergillus flavus as Potential Biocontrol Agents for Aflatoxin in Maize

    USDA-ARS?s Scientific Manuscript database

    Aflatoxin contamination resulting from maize infection by Aspergillus flavus is both an economic concern and public health concern. Therefore, strategies for controlling maize contamination are being investigated. Abilities of 11 naturally occurring atoxigenic strains in Nigeria to reduce aflatox...

  9. Requirement of Simultaneous Assessment of Crystal- and Supernatant-Related Entomotoxic Activities of Bacillus thuringiensis Strains for Biocontrol-Product Development

    PubMed Central

    Argôlo-Filho, Ronaldo Costa; Costa, Robson Luz; Pinheiro, Daniele Heloisa; Corrêa, Fábio Mathias; Valicente, Fernando Hercos; Pomella, Alan William Vilela; Loguercio, Leandro Lopes

    2014-01-01

    Bioinsecticides with lower concentrations of endospores/crystals and without loss of efficiency are economically advantageous for pest biocontrol. In addition to Cry proteins, other Bacillus thuringiensis (Bt) toxins in culture supernatants (SN) have biocontrol potential (e.g., Vip3A, Cry1I, Sip1), whereas others are unwanted (β-exotoxins), as they display widespread toxicity across taxa. A strain simultaneously providing distinct toxin activities in crystals and SN would be desirable for bioinsecticides development; however, strains secreting β-exotoxins should be discarded, independently of other useful entomotoxins. Entomotoxicity of crystals and SN from a Brazilian Bt tolworthi strain (Btt01) was tested against Spodoptera frugiperda to assess the potential for biocontrol-product development based on more than one type of toxin/activity. Tests showed that 107 endospores mL−1 caused >80% of larvae mortality, suggesting Btt01 may be used in similar concentrations as those of other Bt-based biopesticides. When it was applied to cornfields, a significant 60% reduction of larvae infestation was observed. However, bioassays with Btt01 SN revealed a thermostable toxic activity. Physicochemical characterization strongly suggests the presence of unwanted β-exotoxins, with isolate-specific temporal variation in its secretion. Knowledge of the temporal pattern of secretion/activity in culture for all forms of toxins produced by a single strain is required to both detect useful activities and avoid the potential lack of identification of undesirable toxins. These findings are discussed in the contexts of commercial Bt product development, advantages of multiple-activity strains, and care and handling recommended for large-scale fermentation systems. PMID:24854738

  10. Requirement of simultaneous assessment of crystal- and supernatant-related entomotoxic activities of Bacillus thuringiensis strains for biocontrol-product development.

    PubMed

    Argôlo-Filho, Ronaldo Costa; Costa, Robson Luz; Pinheiro, Daniele Heloisa; Corrêa, Fábio Mathias; Valicente, Fernando Hercos; Pomella, Alan William Vilela; Loguercio, Leandro Lopes

    2014-05-20

    Bioinsecticides with lower concentrations of endospores/crystals and without loss of efficiency are economically advantageous for pest biocontrol. In addition to Cry proteins, other Bacillus thuringiensis (Bt) toxins in culture supernatants (SN) have biocontrol potential (e.g., Vip3A, Cry1I, Sip1), whereas others are unwanted (β-exotoxins), as they display widespread toxicity across taxa. A strain simultaneously providing distinct toxin activities in crystals and SN would be desirable for bioinsecticides development; however, strains secreting β-exotoxins should be discarded, independently of other useful entomotoxins. Entomotoxicity of crystals and SN from a Brazilian Bt tolworthi strain (Btt01) was tested against Spodoptera frugiperda to assess the potential for biocontrol-product development based on more than one type of toxin/activity. Tests showed that 10(7) endospores mL(-1) caused >80% of larvae mortality, suggesting Btt01 may be used in similar concentrations as those of other Bt-based biopesticides. When it was applied to cornfields, a significant 60% reduction of larvae infestation was observed. However, bioassays with Btt01 SN revealed a thermostable toxic activity. Physicochemical characterization strongly suggests the presence of unwanted β-exotoxins, with isolate-specific temporal variation in its secretion. Knowledge of the temporal pattern of secretion/activity in culture for all forms of toxins produced by a single strain is required to both detect useful activities and avoid the potential lack of identification of undesirable toxins. These findings are discussed in the contexts of commercial Bt product development, advantages of multiple-activity strains, and care and handling recommended for large-scale fermentation systems.

  11. Evaluation of Pseudomonas syringae Strain ESC11 for Biocontrol of Crown Rot and Anthracnose of Banana

    USDA-ARS?s Scientific Manuscript database

    Pseudomonas syringae strain ESC11, and 250 'g/ml each of thiabendazole (TBZ) and imazalil reduced crown rot of banana caused by a Fusarium sp. by 0-88% and 73-88%, respectively, in laboratory experiments. ESC11 alone did not significantly reduce rot, mold, or anthracnose in most field trials. TBZ an...

  12. Antibiosis activity of Pantoea agglomerans biocontrol strain E325 against Erwinia amylovora on apple flower stigmas.

    PubMed

    Pusey, P L; Stockwell, V O; Reardon, C L; Smits, T H M; Duffy, B

    2011-10-01

    Pantoea agglomerans E325, the active ingredient in a commercial product for fire blight control, was previously shown in vitro to produce a unique alkaline- and phosphate-sensitive antibiotic specific to Erwinia amylovora. Antibiosis was evaluated as a mode of antagonism on flower stigmas using two antibiosis-deficient mutants. On King's medium B, mutants E325ad1 and E325ad2 have stable smooth-butyrous or hypermucoid colony morphologies, respectively, and the parental strain E325 exhibits phenotypic plasticity with predominantly hypermucoid colonies accompanied by slower-growing, smooth-butyrous colonies. Mutants were tested against E. amylovora on stigmas of detached flowers of crab apple (Malus mandshurica) in growth chambers and apple (Malus domestica) in the orchard. Epiphytic fitness of the antibiosis-negative mutants was similar or greater than the parental strain as determined by relative area under the population curve (RAUPC). In laboratory and orchard trials, both mutants had significantly lower inhibitory activity against the pathogen (i.e., less reduction of E. amylovora RAUPC) compared with the parental strain. E325 and the mutants caused similar decreases in pH in a broth medium, indicating that acidification, which was previously reported as a possible mechanism of pathogen inhibition on stigmas, is not directly related to antibiosis. In this study we provide the first evidence for E325 antibiosis involved in E. amylovora growth suppression on apple flower stigmas.

  13. A Perilipin Gene from Clonostachys rosea f. Catenulata HL-1-1 Is Related to Sclerotial Parasitism

    PubMed Central

    Sun, Zhan-Bin; Li, Shi-Dong; Zhong, Zeng-Ming; Sun, Man-Hong

    2015-01-01

    Clonostachys rosea f. catenulata is a promising biocontrol agent against many fungal plant pathogens. To identify mycoparasitism-related genes from C. rosea f. catenulata, a suppression subtractive hybridization (SSH) cDNA library of C. rosea f. catenulata HL-1-1 that parasitizes the sclerotia of S. sclerotiorum was constructed. 502 clones were sequenced randomly, and thereby 472 expressed sequence tags (ESTs) were identified. Forty-three unigenes were annotated and exhibited similarity to a wide diversity of genes. Quantitative real -time PCR showed that a perilipin-like protein encoding gene, Per3, was up-regulated by 6.6-fold over the control at 96 h under the induction of sclerotia. The full-length sequence of Per3 was obtained via 5' and 3' rapid identification of cDNA ends. Overexpression of Per3 in HL-1-1 significantly enhanced the parasitic ability on sclerotia. The results indicated that Per3 might be involved in the mycoparasitism of C. rosea f. catenulata HL-1-1. This is the first report of a perilipin as a potential biocontrol gene in mycoparasites. The study provides usefu l insights into the interaction between C. rosea f. catenulata and fungal plant pathogens. PMID:25761240

  14. Development of a strain-specific genomic marker for monitoring a Bacillus subtilis biocontrol strain in the rhizosphere of tomato.

    PubMed

    Felici, Cristiana; Vettori, Lorenzo; Toffanin, Annita; Nuti, Marco

    2008-08-01

    A strain-specific molecular marker enabling the detection and tracking of the biological control agent Bacillus subtilis 101, when released into the environment, was developed. Random amplified polymorphic DNA (RAPD) technique was used to differentiate this from other B. subtilis strains. A differentially amplified fragment obtained from RAPD profiles was sequenced and characterized as sequence-characterized amplified region (SCAR) marker, and four primer pairs were designed and evaluated for their specificity towards this strain. The sensibility of the selected SCAR primer pair was evaluated by qualitative PCR and Southern blotting, and the detection limit was assessed around 10(2) CFU (g dry wt soil)(-1), thus providing a reliable tool for the traceability of this B. subtilis strain in greenhouse or field trials. A plating assay coupled to PCR with the SCAR primer pair was then used as a detection method in microcosm experiments for monitoring the population of B. subtilis 101 in the rhizosphere of tomato, grown under two different soil conditions, i.e. nonsterile peat-based substrate and sandy-loam agricultural soil, respectively. The data of rhizosphere colonization indicated that the soil conditions significantly affected the rhizosphere establishment of strain 101.

  15. Biocontrol of Monilinia laxa by Aureobasidium pullulans strains: Insights on competition for nutrients and space.

    PubMed

    Di Francesco, Alessandra; Ugolini, Luisa; D'Aquino, Salvatore; Pagnotta, Eleonora; Mari, Marta

    2017-02-17

    Two Aureobasidium pullulans strains (L1 and L8), able to prevent postharvest fruit decay, were evaluated in order to elucidate how the competition for nutrients and space was involved in their activity against Monilinia laxa, the causal agent of peach brown rot. The competition for nutrients was studied by co-culturing pathogen conidia and antagonists in different conditions of nutrient availability and avoiding contact between them. Both antagonists prevented the germination of conidia of M. laxa in water, reducing germination rate by >35%. However, L1 and L8 showed the lowest inhibition of conidial germination in peach juice at 5%, with a reduction of 12.6% and 13.9% respectively. HPLC amino acid analysis of peach juice revealed that the addition of the yeast suspension greatly modified their composition: asparagine was completely depleted soon after 12h of incubation and was probably hydrolyzed to aspartic acid by the yeasts, as aspartic acid content markedly increased. Pure asparagine and aspartic acid were tested by in vitro trials at the concentrations found in peach juice: both influenced M. laxa growth, but in opposite ways. Asparagine stimulated pathogen growth; conversely, amended medium with aspartic acid significantly inhibited the conidia germination and mycelial development of M. laxa. Scanning Electron Microscopy revealed that both strains showed a great capability to compete with M. laxa for space (starting 8h after treatment), colonizing the wound surface and inhibiting pathogen growth. This study clearly showed that A. pullulans L1 and L8 strains could compete with M. laxa for nutrients and space; this mode of action may play an important role in the antagonistic activity, especially in the first hours of tritrophic host-pathogen-antagonist interaction, although several other mechanisms can interact each other.

  16. Genome Sequencing of a Mung Bean Plant Growth Promoting Strain of P. aeruginosa with Biocontrol Ability

    PubMed Central

    Illakkiam, Devaraj; Shankar, Manoharan; Ponraj, Paramasivan; Rajendhran, Jeyaprakash

    2014-01-01

    Pseudomonas aeruginosa PGPR2 is a mung bean rhizosphere strain that produces secondary metabolites and hydrolytic enzymes contributing to excellent antifungal activity against Macrophomina phaseolina, one of the prevalent fungal pathogens of mung bean. Genome sequencing was performed using the Ion Torrent Personal Genome Machine generating 1,354,732 reads (6,772,433 sequenced bases) achieving ~25-fold coverage of the genome. Reference genome assembly using MIRA 3.4.0 yielded 198 contigs. The draft genome of PGPR2 encoded 6803 open reading frames, of which 5314 were genes with predicted functions, 1489 were genes of known functions, and 80 were RNA-coding genes. Strain specific and core genes of P. aeruginosa PGPR2 that are relevant to rhizospheric habitat were identified by pangenome analysis. Genes involved in plant growth promoting function such as synthesis of ACC deaminase, indole-3-acetic acid, trehalose, mineral scavenging siderophores, hydrogen cyanide, chitinases, acyl homoserine lactones, acetoin, 2,3-butanediol, and phytases were identified. In addition, niche-specific genes such as phosphate solubilising 3-phytase, adhesins, pathway-specific transcriptional regulators, a diguanylate cyclase involved in cellulose synthesis, a receptor for ferrienterochelin, a DEAD/DEAH-box helicase involved in stress tolerance, chemotaxis/motility determinants, an HtpX protease, and enzymes involved in the production of a chromanone derivative with potent antifungal activity were identified. PMID:25184130

  17. A novel strain of Brevibacillus laterosporus produces chitinases that contribute to its biocontrol potential.

    PubMed

    Prasanna, Lakshmi; Eijsink, Vincent G H; Meadow, Richard; Gåseidnes, Sigrid

    2013-02-01

    A novel strain exhibiting entomopathogenic and chitinolytic activity was isolated from mangrove marsh soil in India. The isolate was identified as Brevibacillus laterosporus by phenotypic characterization and 16S rRNA sequencing and designated Lak1210. When grown in the presence of colloidal chitin as the sole carbon source, the isolate produced extracellular chitinases. Chitinase activity was inhibited by allosamidin indicating that the enzymes belong to the family 18 chitinases. The chitinases were purified by ammonium sulfate precipitation followed by chitin affinity chromatography yielding chitinases and chitinase fragments with 90, 75, 70, 55, 45, and 25 kDa masses. Mass spectrometric analyses of tryptic fragments showed that these fragments belong to two distinct chitinases that are almost identical to two putative chitinases, a 89.6-kDa four-domain chitodextrinase and a 69.4-kDa two-domain enzyme called ChiA1, that are encoded on the recently sequenced genome of B. laterosporus LMG15441. The chitinase mixture showed two pH optima, at 6.0 and 8.0, and an optimum temperature of 70 °C. The enzymes exhibited antifungal activity against the phytopathogenic fungus Fusarium equiseti. Insect toxicity bioassays with larvae of diamondback moths (Plutella xylostella), showed that addition of chitinases reduced the time to reach 50 % mortality upon infection with non-induced B. laterosporus from 3.3 to 2.1 days. This study provides evidence for the presence of inducible, extracellular chitinolytic enzymes in B. laterosporus that contribute to the strain's antifungal activity and insecticidal activity.

  18. An endophytic Streptomyces sp. strain DHV3-2 from diseased root as a potential biocontrol agent against Verticillium dahliae and growth elicitor in tomato (Solanum lycopersicum).

    PubMed

    Cao, Peng; Liu, Chongxi; Sun, Pengyu; Fu, Xuepeng; Wang, Shaoxian; Wu, Fengzhi; Wang, Xiangjing

    2016-12-01

    Plant endophytes play important roles in biocontrol of plant diseases. Actinomycetes are used for biocontrol of fungal diseases caused by Verticillium dahliae. Many studies have focused on the endophytic actinomycetes isolated from the roots of healthy plants, but few on those from the roots of diseased plants. In the present research, actinomycetes were isolated from the roots of diseased and healthy tomato plants, respectively. The results showed that, in total, 86 endophytic actinomycetes were isolated for screening of their antimicrobial activities, 8 of which showed antagonism to V. dahliae in vitro. Among the 8 antagonistic strains, 5 (out of 36) were from the roots of diseased plants, with inhibition diameter zones ranging from 11.2 to 18.2 mm, whereas 3 (out of 50) were from the roots of healthy plants, with inhibition diameter zones ranging from 11.5 to 15.5 mm. Endophytic strain DHV3-2 was isolated from the root of a diseased plant and demonstrated a potent effect against V. dahliae and other pathogenic fungi by showing the largest inhibition diameter zones among all the eight antagonistic strains. Thus, strain DHV3-2 was chosen to investigate its biological control efficacies in vivo. Further study showed that the disease incidence and disease severity indices of tomato Verticillium wilt decreased significantly (P < 0.05). We also found that the plant shoot fresh weight and height increased greatly (P < 0.05) upon treatment with strain DHV3-2 compared to the plants uninoculated in greenhouse conditions. Root colonization showed that strain DHV3-2 had the higher root-colonizing capacity in the roots of infected plants compared with the roots of healthy plants. This isolate was identified as Streptomyces sp. based on morphological characteristics and 16S rRNA gene analysis. In conclusion, the roots of diseased tomato plants are a potential reservoir of biological control actinomycetes, and Streptomyces sp. strain DHV3-2 is a potential biocontrol

  19. A phenazine-1-carboxylic acid producing polyextremophilic Pseudomonas chlororaphis (MCC2693) strain, isolated from mountain ecosystem, possesses biocontrol and plant growth promotion abilities.

    PubMed

    Jain, Rahul; Pandey, Anita

    2016-09-01

    The genus Pseudomonas is known to comprise a huge diversity of species with the ability to thrive in different habitats, including those considered as extreme environments. In the present study, a psychrotolerant, wide pH tolerant and halotolerant strain of Pseudomonas chlororaphis GBPI_507 (MCC2693), isolated from the wheat rhizosphere growing in a mountain location in Indian Himalayan Region (IHR), has been investigated for its antimicrobial potential with particular reference to phenazine production and plant growth promoting traits. GBPI_507 showed phenazine production at the temperatures ranged from 14 to 25°C. The benzene extracted compound identified as phenazine-1-carboxylic acid (PCA) through GC-MS exhibited antimicrobial properties against Gram positive bacteria and actinomycetes. The inhibition of phytopathogens in diffusible biocontrol assays was recorded in an order: Alternaria alternata>Phytophthora sp.>Fusarium solani>F. oxysporum. In volatile metabolite assays, all the pathogens, except Phytophthora sp. produced distorted colonies, characterized by restricted sporulation. The isolate also possessed other growth promoting and biocontrol traits including phosphate solubilization and production of siderophores, HCN, ammonia, and lytic enzymes (lipase and protease). Molecular studies confirmed production of PCA by the bacterium GBPI_507 through presence of phzCD and phzE genes in its genome. The polyextremophilic bacterial strain possesses various important characters to consider it as a potential agent for field applications, especially in mountain ecosystem, for sustainable and eco-friendly crop production. Copyright © 2016 Elsevier GmbH. All rights reserved.

  20. Soil Type Dependent Rhizosphere Competence and Biocontrol of Two Bacterial Inoculant Strains and Their Effects on the Rhizosphere Microbial Community of Field-Grown Lettuce

    PubMed Central

    Schreiter, Susanne; Sandmann, Martin; Smalla, Kornelia; Grosch, Rita

    2014-01-01

    Rhizosphere competence of bacterial inoculants is assumed to be important for successful biocontrol. Knowledge of factors influencing rhizosphere competence under field conditions is largely lacking. The present study is aimed to unravel the effects of soil types on the rhizosphere competence and biocontrol activity of the two inoculant strains Pseudomonas jessenii RU47 and Serratia plymuthica 3Re4-18 in field-grown lettuce in soils inoculated with Rhizoctonia solani AG1-IB or not. Two independent experiments were carried out in 2011 on an experimental plot system with three soil types sharing the same cropping history and weather conditions for more than 10 years. Rifampicin resistant mutants of the inoculants were used to evaluate their colonization in the rhizosphere of lettuce. The rhizosphere bacterial community structure was analyzed by denaturing gradient gel electrophoresis of 16S rRNA gene fragments amplified from total community DNA to get insights into the effects of the inoculants and R. solani on the indigenous rhizosphere bacterial communities. Both inoculants showed a good colonization ability of the rhizosphere of lettuce with more than 106 colony forming units per g root dry mass two weeks after planting. An effect of the soil type on rhizosphere competence was observed for 3Re4-18 but not for RU47. In both experiments a comparable rhizosphere competence was observed and in the presence of the inoculants disease symptoms were either significantly reduced, or at least a non-significant trend was shown. Disease severity was highest in diluvial sand followed by alluvial loam and loess loam suggesting that the soil types differed in their conduciveness for bottom rot disease. Compared to effect of the soil type of the rhizosphere bacterial communities, the effects of the pathogen and the inoculants were less pronounced. The soil types had a surprisingly low influence on rhizosphere competence and biocontrol activity while they significantly affected

  1. Soil type dependent rhizosphere competence and biocontrol of two bacterial inoculant strains and their effects on the rhizosphere microbial community of field-grown lettuce.

    PubMed

    Schreiter, Susanne; Sandmann, Martin; Smalla, Kornelia; Grosch, Rita

    2014-01-01

    Rhizosphere competence of bacterial inoculants is assumed to be important for successful biocontrol. Knowledge of factors influencing rhizosphere competence under field conditions is largely lacking. The present study is aimed to unravel the effects of soil types on the rhizosphere competence and biocontrol activity of the two inoculant strains Pseudomonas jessenii RU47 and Serratia plymuthica 3Re4-18 in field-grown lettuce in soils inoculated with Rhizoctonia solani AG1-IB or not. Two independent experiments were carried out in 2011 on an experimental plot system with three soil types sharing the same cropping history and weather conditions for more than 10 years. Rifampicin resistant mutants of the inoculants were used to evaluate their colonization in the rhizosphere of lettuce. The rhizosphere bacterial community structure was analyzed by denaturing gradient gel electrophoresis of 16S rRNA gene fragments amplified from total community DNA to get insights into the effects of the inoculants and R. solani on the indigenous rhizosphere bacterial communities. Both inoculants showed a good colonization ability of the rhizosphere of lettuce with more than 10(6) colony forming units per g root dry mass two weeks after planting. An effect of the soil type on rhizosphere competence was observed for 3Re4-18 but not for RU47. In both experiments a comparable rhizosphere competence was observed and in the presence of the inoculants disease symptoms were either significantly reduced, or at least a non-significant trend was shown. Disease severity was highest in diluvial sand followed by alluvial loam and loess loam suggesting that the soil types differed in their conduciveness for bottom rot disease. Compared to effect of the soil type of the rhizosphere bacterial communities, the effects of the pathogen and the inoculants were less pronounced. The soil types had a surprisingly low influence on rhizosphere competence and biocontrol activity while they significantly affected

  2. Characterization and evaluation of Bacillus amyloliquefaciens strain WF02 regarding its biocontrol activities and genetic responses against bacterial wilt in two different resistant tomato cultivars.

    PubMed

    Huang, Chu-Ning; Lin, Chan-Pin; Hsieh, Feng-Chia; Lee, Sook-Kuan; Cheng, Kuan-Chen; Liu, Chi-Te

    2016-11-01

    Bacillus amyloliquefaciens strain WF02, isolated from soil collected at Wufeng Mountain, Taiwan, has siderophore-producing ability and in vitro antagonistic activity against bacterial wilt pathogen. To determine the impact of plant genotype on biocontrol effectiveness, we treated soil with this strain before infecting susceptible (L390) and moderately resistant (Micro-Tom) tomato cultivars with Ralstonia solanacearum strain Pss4. We also compared the efficacy of this strain with that of commercial Bacillus subtilis strain Y1336. Strain WF02 provided longer lasting protection against R. solanacearum than did strain Y1336 and controlled the development of wilt in both cultivars. To elucidate the genetic responses in these plants under WF02 treatment, we analyzed the temporal expression of defense-related genes in leaves. The salicylic acid pathway-related genes phenylalanine ammonia-lyase and pathogenesis-related protein 1a were up-regulated in both cultivars, whereas expression of the jasmonic acid pathway-related gene lipoxygenase was only elevated in the susceptible tomato cultivar (L390). These results suggest that WF02 can provide protection against bacterial wilt in tomato cultivars with different levels of disease resistance via direct and indirect modes of action.

  3. The development of genetic and molecular markers to register and commercialize Penicillium rubens (formerly Penicillium oxalicum) strain 212 as a biocontrol agent.

    PubMed

    Villarino, Maria; De Cal, Antonieta; Melgarejo, Paloma; Larena, Inmaculada; Espeso, Eduardo A

    2016-01-01

    Penicillium oxalicum strain 212 (PO212) is an effective biocontrol agent (BCA) against a large number of economically important fungal plant pathogens. For successful registration as a BCA in Europe, PO212 must be accurately identified. In this report, we describe the use of classical genetic and molecular markers to characterize and identify PO212 in order to understand its ecological role in the environment or host. We successfully generated pyrimidine (pyr-) auxotrophic mutants. In addition we also designed specific oligonucleotides for the pyrF gene at their untranslated regions for rapid and reliable identification and classification of strains of P. oxalicum and P. rubens, formerly P. chrysogenum. Using these DNA-based technologies, we found that PO212 is a strain of P. rubens, and is not a strain of P. oxalicum. This work presents PO212 as the unique P. rubens strain to be described as a BCA and the information contained here serves for its registration and commercialization in Europe. © 2015 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  4. Biocontrol activity of four non- and low-fermenting yeast strains against Aspergillus carbonarius and their ability to remove ochratoxin A from grape juice.

    PubMed

    Fiori, Stefano; Urgeghe, Pietro Paolo; Hammami, Walid; Razzu, Salvatorico; Jaoua, Samir; Migheli, Quirico

    2014-10-17

    Aspergillus spp. infection of grape may lead to ochratoxin A (OTA) contamination in processed beverages such as wine and grape juice. The aim of the current study was to evaluate the biocontrol potential of two non-fermenting (Cyberlindnera jadinii 273 and Candida friedrichii 778) and two low-fermenting (Candida intermedia 235 and Lachancea thermotolerans 751) yeast strains against the pathogenic fungus and OTA-producer Aspergillus carbonarius, and their ability to remove OTA from grape juice. Two strains, 235 and 751, showed a significant ability to inhibit A. carbonarius both on grape berries and in in vitro experiments. Neither their filtrate nor their autoclaved filtrate culture broth was able to prevent consistently pathogen growth. Volatile organic compounds (VOCs) produced by all four selected yeasts were likely able to consistently prevent pathogen sporulation in vitro. VOCs produced by the non-fermenting strain 778 also significantly reduced A. carbonarius vegetative growth. Three yeast strains (235, 751, and 778) efficiently adsorbed artificially spiked OTA from grape juice, while autoclaving treatment improved OTA adsorption capacity by all the four tested strains. Biological control of A. carbonarius and OTA-decontamination using yeast is proposed as an approach to meet the Islamic dietary laws concerning the absence of alcohol in halal beverages.

  5. Genomic and phenotypic characterization of a nonpigmented variant of Pantoea vagans biocontrol strain C9-1 lacking the 530-kb megaplasmid pPag3.

    PubMed

    Smits, Theo H M; Rezzonico, Fabio; Pelludat, Cosima; Goesmann, Alexander; Frey, Jürg E; Duffy, Brion

    2010-07-01

    A 530-kb megaplasmid pPag3 contributing 10.8% of the total genome of Pantoea vagans biocontrol strain C9-1 was sequenced. A rare nonpigmented variant C9-1W was obtained and shown to have lost pPag3, but retained all other plasmids (pPag1, pPag2). Phenotypic characterization of the variant confirmed the function of several annotated genes that may influence ecological fitness and efficacy. Metabolic profiling revealed important plasmid-based carbon utilization phenotypes. Plasmid loss resulted in thiamine auxotrophy, absence of carotenoid pigmentation, desferrioxamine diffusible siderophore biosynthesis, inherent ampicillin resistance and expression of AI-1 quorum-sensing signaling. This confirmed the functional expression of the corresponding genes located on pPag3 in P. vagans.

  6. Effect of organic fertilizers prepared from organic waste materials on the production of antibacterial volatile organic compounds by two biocontrol Bacillus amyloliquefaciens strains.

    PubMed

    Raza, Waseem; Wei, Zhong; Ling, Ning; Huang, Qiwei; Shen, Qirong

    2016-06-10

    Three organic fertilizers made of different animal and plant waste materials (BOFs) were evaluated for their effects on the production of antibacterial volatile organic compounds (VOCs) by two Bacillus amyloliquefaciens strains SQR-9 and T-5 against the tomato wilt pathogen Ralstonia solanacearum (RS). Both strains could produce VOCs that inhibited the growth and virulence traits of RS; however, in the presence of BOFs, the production of antibacterial VOCs was significantly increased. The maximum inhibition of growth and virulence traits of RS by VOCs of T-5 and SQR-9 was determined at 1.5% BOF2 and 2% BOF3, respectively. In case of strain T-5, 2-nonanone, nonanal, xylene, benzothiazole, and butylated hydroxy toluene and in case of strain SQR-9, 2-nonanone, nonanal, xylene and 2-undecanone were the main antibacterial VOCs whose production was increased in the presence of BOFs. The results of this study reveal another significance of using organic fertilizers to improve the antagonistic activity of biocontrol agents against phytopathogens.

  7. Selection of reliable reference genes for gene expression studies in Clonostachys rosea 67-1 under sclerotial induction.

    PubMed

    Sun, Zhan-Bin; Li, Shi-Dong; Sun, Man-Hong

    2015-07-01

    Reference genes are important to precisely quantify gene expression by real-time PCR. In order to identify stable and reliable expressed genes in mycoparasite Clonostachys rosea in different modes of nutrition, seven commonly used housekeeping genes, 18S rRNA, actin, β-tubulin, elongation factor 1, ubiquitin, ubiquitin-conjugating enzyme and glyceraldehyde-3-phosphate dehydrogenase, from the effective biocontrol isolate C. rosea 67-1 were tested for their expression under sclerotial induction and during vegetative growth on PDA medium. Analysis by three software programs showed that differences existed among the candidates. Elongation factor 1 was most stable; the M value in geNorm, SD value in Bestkeeper and stability value in Normfinder analysis were 0.405, 0.450 and 0.442, respectively, indicating that the gene elongation factor 1 could be used to normalize gene expression in C. rosea in both vegetative growth and parasitic process. By using elongation factor 1, the expression of a serine protease gene, sep, in different conditions was assessed, which was consistent with the transcriptomic data. This research provides an effective method to quantitate expression changes of target genes in C. rosea, and will assist in further investigation of parasitism-related genes of this fungus.

  8. The roles of inoculants' carbon source use in the biocontrol of potato scab disease.

    PubMed

    Sun, Pingping; Zhao, Xinbei; Shangguan, Nini; Chang, Dongwei; Ma, Qing

    2015-04-01

    Despite the application of multiple strains in the biocontrol of plant diseases, multistrain inoculation is still constrained by its inconsistency in the field. Nutrients, especially carbons, play an important role in the biocontrol processes. However, little work has been done on the systematic estimation of inoculants' carbon source use on biocontrol efficacies in vivo. In the present study, 7 nonpathogenic Streptomyces strains alone and in different combinations were inoculated as biocontrol agents against the potato scab disease, under field conditions and greenhouse treatments. The influence of the inoculants' carbon source use properties on biocontrol efficacies was investigated. The results showed that increasing the number of inoculated strains did not necessarily result in greater biocontrol efficacy in vivo. However, single strains with higher growth rates or multiple strains with less carbon source competition had positive effects on the biocontrol efficacies. These findings may shed light on optimizing the consistent biocontrol of plant disease with the consideration of inoculants' carbon source use properties.

  9. Effects of community versus single strain inoculants on the biocontrol of Salmonella and microbial community dynamics in alfalfa sprouts.

    PubMed

    Matos, Anabelle; Garland, Jay L

    2005-01-01

    Potential biological control inoculants, Pseudomonas fluorescens 2-79 and microbial communities derived from market sprouts or laboratory-grown alfalfa sprouts, were introduced into alfalfa seeds with and without a Salmonella inoculum. We examined their ability to inhibit the growth of this foodborne pathogen and assess the relative effects of the inoculants on the alfalfa microbial community structure and function. Alfalfa seeds contaminated with a Salmonella cocktail were soaked for 2 h in bacterial suspensions from each inoculant tested. Inoculated alfalfa seeds were grown for 7 days and sampled during days 1, 3, and 7. At each sampling, alfalfa sprouts were sonicated for 7 min to recover microflora from the surface, and the resulting suspensions were diluted and plated on selective and nonselective media. Total bacterial counts were obtained using acridine orange staining, and the percentage culturability was calculated. Phenotypic potential of sprout-associated microbial communities inoculated with biocontrol treatments was assessed using community-level physiological profiles based on patterns of use of 95 separate carbon sources in Biolog plates. Community-level physiological profiles were also determined using oxygen-sensitive fluorophore in BD microtiter plates to examine functional patterns in these communities. No significant differences in total and mesophilic aerobe microbial cell density or microbial richness resulting from the introduction of inoculants on alfalfa seeds with and without Salmonella were observed. P. fluorescens 2-79 exhibited the greatest reduction in the growth of Salmonella early during alfalfa growth (4.22 log at day 1), while the market sprout inoculum had the reverse effect, resulting in a maximum log reduction (5.48) of Salmonella on day 7. Community-level physiological profiles analyses revealed that market sprout communities peaked higher and faster compared with the other inoculants tested. These results suggest that

  10. Biocontrol Potential of Streptomyces hydrogenans Strain DH16 toward Alternaria brassicicola to Control Damping Off and Black Leaf Spot of Raphanus sativus.

    PubMed

    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.

  11. 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

  12. Potential of Epicoccum purpurascens Strain 5615 AUMC as a Biocontrol Agent of Pythium irregulare Root Rot in Three Leguminous Plants

    PubMed Central

    Koutb, Mostafa

    2010-01-01

    Epicoccum purpurascens stain 5615 AUMC was investigated for its biocontrol activity against root rot disease caused by Pythium irregulare. E. purpurascens greenhouse pathogenicity tests using three leguminous plants indicated that the fungus was nonpathogenic under the test conditions. The germination rate of the three species of legume seeds treated with a E. purpurascens homogenate increased significantly compared with the seeds infested with P. irregulare. No root rot symptoms were observed on seeds treated with E. purpurascens, and seedlings appeared more vigorous when compared with the non-treated control. A significant increase in seedling growth parameters (seedling length and fresh and dry weights) was observed in seedlings treated with E. purpurascens compared to pathogen-treated seedlings. Pre-treating the seeds with the bioagent fungus was more efficient for protecting seeds against the root rot disease caused by P. irregulare than waiting for disease dispersal before intervention. To determine whether E. purpurascens produced known anti-fungal compounds, an acetone extract of the fungus was analyzed by gas chromatography mass spectrometry. The extract revealed a high percentage of the cinnamic acid derivative (trimethylsiloxy) cinnamic acid methyl ester. The E. purpurascens isolate grew more rapidly than the P. irregulare pathogen in a dual culture on potato dextrose agar nutrient medium, although the two fungi grew similarly when cultured separately. This result may indicate antagonism via antibiosis or competition. PMID:23956668

  13. Biocontrol of Fusarium Crown and Root Rot and Promotion of Growth of Tomato by Paenibacillus Strains Isolated from Soil

    PubMed Central

    Xu, Sheng Jun

    2014-01-01

    In this study, bacterial strains were isolated from soils from 30 locations of Samcheok, Gangwon province. Of the isolated strains, seven showed potential plant growth promoting and antagonistic activities. Based on cultural and morphological characterization, and 16S rRNA gene sequencing, these strains were identified as Paenibacillus species. All seven strains produced ammonia, cellulase, hydrocyanic acid, indole-3-acetic acid, protease, phosphatase, and siderophores. They also inhibited the mycelial growth of Fusarium oxysporum f. sp. radicis-lycopersici in vitro. The seven Paenibacillus strains enhanced a range of growth parameters in tomato plants under greenhouse conditions, in comparison with non-inoculated control plants. Notably, treatment of tomato plants with one identified strain, P. polymyxa SC09-21, resulted in 80.0% suppression of fusarium crown and root rot under greenhouse conditions. The plant growth promoting and antifungal activity of P. polymyxa SC09-21 identified in this study highlight its potential suitability as a bioinoculant. PMID:25071385

  14. The plant pathogenic fungus Gaeumannomyces graminis var. tritici improves bacterial growth and triggers early gene regulations in the biocontrol strain Pseudomonas fluorescens Pf29Arp.

    PubMed

    Barret, M; Frey-Klett, P; Boutin, M; Guillerm-Erckelboudt, A-Y; Martin, F; Guillot, L; Sarniguet, A

    2009-01-01

    In soil, some antagonistic rhizobacteria contribute to reduce root diseases caused by phytopathogenic fungi. Direct modes of action of these bacteria have been largely explored; however, commensal interaction also takes place between these microorganisms and little is known about the influence of filamentous fungi on bacteria. An in vitro confrontation bioassay between the pathogenic fungus Gaeumannomyces graminis var. tritici (Ggt) and the biocontrol bacterial strain Pseudomonas fluorescens Pf29Arp was set up to analyse bacterial transcriptional changes induced by the fungal mycelium at three time-points of the interaction before cell contact and up until contact. For this, a Pf29Arp shotgun DNA microarray was constructed. Specifity of Ggt effect was assessed in comparison with one of two other filamentous fungi, Laccaria bicolor and Magnaporthe grisea. During a commensal interaction, Ggt increased the growth rate of Pf29Arp. Before contact, Ggt induced bacterial genes involved in mycelium colonization. At contact, genes encoding protein of stress response and a patatin-like protein were up-regulated. Among all the bacterial genes identified, xseB was specifically up-regulated at contact by Ggt but down-regulated by the other fungi. Data showed that the bacterium sensed the presence of the fungus early, but the main gene alteration occurred during bacterial-fungal cell contact.

  15. Transcriptomic profiling of microbe-microbe interactions reveals the specific response of the biocontrol strain P. fluorescens In5 to the phytopathogen Rhizoctonia solani.

    PubMed

    Hennessy, Rosanna C; Glaring, Mikkel A; Olsson, Stefan; Stougaard, Peter

    2017-08-10

    Few studies to date report the transcriptional response of biocontrol bacteria toward phytopathogens. In order to gain insights into the potential mechanism underlying the antagonism of the antimicrobial producing strain P. fluorescens In5 against the phytopathogens Rhizoctonia solani and Pythium aphanidermatum, global RNA sequencing was performed. Differential gene expression profiling of P. fluorescens In5 in response to either R. solani or P. aphanidermatum was investigated using transcriptome sequencing (RNA-seq). Total RNA was isolated from single bacterial cultures of P. fluorescens In5 or bacterial cultures in dual-culture for 48 h with each pathogen in biological triplicates. RNA-seq libraries were constructed following a default Illumina stranded RNA protocol including rRNA depletion and were sequenced 2 × 100 bases on Illumina HiSeq generating approximately 10 million reads per sample. No significant changes in global gene expression were recorded during dual-culture of P. fluorescens In5 with any of the two pathogens but rather each pathogen appeared to induce expression of a specific set of genes. A particularly strong transcriptional response to R. solani was observed and notably several genes possibly associated with secondary metabolite detoxification and metabolism were highly upregulated in response to the fungus. A total of 23 genes were significantly upregulated and seven genes were significantly downregulated with at least respectively a threefold change in expression level in response to R. solani compared to the no fungus control. In contrast, only one gene was significantly upregulated over threefold and three transcripts were significantly downregulated over threefold in response to P. aphanidermatum. Genes known to be involved in synthesis of secondary metabolites, e.g. non-ribosomal synthetases and hydrogen cyanide were not differentially expressed at the time points studied. This study demonstrates that genes possibly involved in

  16. Use of Lactobacillus plantarum Strains as a Bio-Control Strategy against Food-Borne Pathogenic Microorganisms

    PubMed Central

    Arena, Mattia Pia; Silvain, Amandine; Normanno, Giovanni; Grieco, Francesco; Drider, Djamel; Spano, Giuseppe; Fiocco, Daniela

    2016-01-01

    Lactobacillus plantarum is one of the most versatile species extensively used in the food industry both as microbial starters and probiotic microorganisms. Several L. plantarum strains have been shown to produce different antimicrobial compounds such as organic acids, hydrogen peroxide, diacetyl, and also bacteriocins and antimicrobial peptides, both denoted by a variable spectrum of action. In recent decades, the selection of microbial molecules and/or bacterial strains able to produce antagonistic molecules to be used as antimicrobials and preservatives has been attracting scientific interest, in order to eliminate or reduce chemical additives, because of the growing attention of consumers for healthy and natural food products. The aim of this work was to investigate the antimicrobial activity of several food-isolated L. plantarum strains, analyzed against the pathogenic bacteria Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7 and Staphylococcus aureus. Antagonistic activity was assayed by agar spot test and revealed that strain L. plantarum 105 had the strongest ability to contrast the growth of L. monocytogenes, while strains L. plantarum 106 and 107 were the most active microorganisms against E. coli O157:H7. The antimicrobial ability was also screened by well diffusion assay and broth micro-dilution method using cell-free supernatants (CFS) from each Lactobacillus strain. Moreover, the chemical nature of the molecules released in the CFS, and possibly underlying the antagonistic activity, was preliminary characterized by exposure to different constraints such as pH neutralization, heating, catalase, and proteinase treatments. Our data suggest that the ability of L. plantarum cultures to contrast pathogens growth in vitro depends, at least in part, on a pH-lowering effect of supernatants and/or on the presence of organic acids. Cluster analysis was performed in order to group L. plantarum strains according to their antimicrobial effect

  17. Use of Lactobacillus plantarum Strains as a Bio-Control Strategy against Food-Borne Pathogenic Microorganisms.

    PubMed

    Arena, Mattia Pia; Silvain, Amandine; Normanno, Giovanni; Grieco, Francesco; Drider, Djamel; Spano, Giuseppe; Fiocco, Daniela

    2016-01-01

    Lactobacillus plantarum is one of the most versatile species extensively used in the food industry both as microbial starters and probiotic microorganisms. Several L. plantarum strains have been shown to produce different antimicrobial compounds such as organic acids, hydrogen peroxide, diacetyl, and also bacteriocins and antimicrobial peptides, both denoted by a variable spectrum of action. In recent decades, the selection of microbial molecules and/or bacterial strains able to produce antagonistic molecules to be used as antimicrobials and preservatives has been attracting scientific interest, in order to eliminate or reduce chemical additives, because of the growing attention of consumers for healthy and natural food products. The aim of this work was to investigate the antimicrobial activity of several food-isolated L. plantarum strains, analyzed against the pathogenic bacteria Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7 and Staphylococcus aureus. Antagonistic activity was assayed by agar spot test and revealed that strain L. plantarum 105 had the strongest ability to contrast the growth of L. monocytogenes, while strains L. plantarum 106 and 107 were the most active microorganisms against E. coli O157:H7. The antimicrobial ability was also screened by well diffusion assay and broth micro-dilution method using cell-free supernatants (CFS) from each Lactobacillus strain. Moreover, the chemical nature of the molecules released in the CFS, and possibly underlying the antagonistic activity, was preliminary characterized by exposure to different constraints such as pH neutralization, heating, catalase, and proteinase treatments. Our data suggest that the ability of L. plantarum cultures to contrast pathogens growth in vitro depends, at least in part, on a pH-lowering effect of supernatants and/or on the presence of organic acids. Cluster analysis was performed in order to group L. plantarum strains according to their antimicrobial effect

  18. Evaluation of indigenous bacterial strains for biocontrol of the frogeye leaf spot of soya bean caused by Cercospora sojina.

    PubMed

    Simonetti, E; Carmona, M A; Scandiani, M M; García, A F; Luque, A G; Correa, O S; Balestrasse, K B

    2012-08-01

    Assessment of biological control of Cercospora sojina, causal agent of frogeye leaf spot (FLS) of soya bean, using three indigenous bacterial strains, BNM297 (Pseudomonas fluorescens), BNM340 and BNM122 (Bacillus amyloliquefaciens). From cultures of each bacterial strain, cell suspensions and cell-free supernatants were obtained and assayed to determine their antifungal activity against C. sojina. Both mycelial growth and spore germination in vitro were more strongly inhibited by bacterial cell suspensions than by cell-free supernatants. The Bacillus strains BNM122 and BNM340 inhibited the fungal growth to a similar degree (I ≈ 52-53%), while cells from P. fluorescens BNM297 caused a lesser reduction (I ≈ 32-34%) in the fungus colony diameter. The foliar application of the two Bacillus strains on soya bean seedlings, under greenhouse conditions, significantly reduced the disease severity with respect to control soya bean seedlings and those sprayed with BNM297. This last bacterial strain was not effective in controlling FLS in vivo. Our data demonstrate that the application of antagonistic bacteria may be a promising and environmentally friendly alternative to control the FLS of soya bean.   To our knowledge, this is the first report of biological control of C. sojina by using native Bacillus strains. © 2012 The Authors. Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.

  19. Methods for the Evaluation of the Bioactivity and Biocontrol Potential of Species of Trichoderma.

    PubMed

    Steyaert, Johanna; Hicks, Emily; Kandula, Janaki; Kandula, Diwakar; Alizadeh, Hossein; Braithwaite, Mark; Yardley, Jessica; Mendoza-Mendoza, Artemio

    2016-01-01

    Members of the genus Trichoderma comprise the majority of commercial fungal biocontrol agents of plant diseases. As such, there is a wealth of information available on the analysis of their biocontrol potential and the mechanisms behind their superior abilities. This chapter aims to summarize the most common methods utilized within a Trichoderma biocontrol program for assessing the biological properties of individual strains.

  20. Prevalence of type III secretion system in effective biocontrol pseudomonads.

    PubMed

    Almario, Juliana; Gobbin, Davide; Défago, Geneviève; Moënne-Loccoz, Yvan; Rezzonico, Fabio

    2014-05-01

    Functional type III secretion system (T3SS) genes are needed for effective biocontrol of Pythium damping-off of cucumber by Pseudomonas fluorescens KD, but whether biocontrol Pseudomonas strains with T3SS genes display overall a higher plant-protecting activity is unknown. The assessment of 198 biocontrol fluorescent pseudomonads originating from 60 soils worldwide indicated that 32% harbour the ATPase-encoding T3SS gene hrcN, which was most often found in tomato isolates. The hrcN(+) biocontrol strains (and especially those also producing 2,4-diacetylphloroglucinol and displaying 1-aminocyclopropane-1-carboxylate deaminase activity) displayed higher plant-protecting ability in comparison with hrcN(-) biocontrol strains, both in the Pythium/cucumber and Fusarium/cucumber pathosystems. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  1. Molecular Characterization and Identification of Biocontrol Isolates of Trichoderma spp.

    PubMed Central

    Hermosa, M. R.; Grondona, I.; Iturriaga, E. A.; Diaz-Minguez, J. M.; Castro, C.; Monte, E.; Garcia-Acha, I.

    2000-01-01

    The most common biological control agents (BCAs) of the genus Trichoderma have been reported to be strains of Trichoderma virens, T. harzianum, and T. viride. Since Trichoderma BCAs use different mechanisms of biocontrol, it is very important to explore the synergistic effects expressed by different genotypes for their practical use in agriculture. Characterization of 16 biocontrol strains, previously identified as “Trichoderma harzianum” Rifai and one biocontrol strain recognized as T. viride, was carried out using several molecular techniques. A certain degree of polymorphism was detected in hybridizations using a probe of mitochondrial DNA. Sequencing of internal transcribed spacers 1 and 2 (ITS1 and ITS2) revealed three different ITS lengths and four different sequence types. Phylogenetic analysis based on ITS1 sequences, including type strains of different species, clustered the 17 biocontrol strains into four groups: T. harzianum-T. inhamatum complex, T. longibrachiatum, T. asperellum, and T. atroviride-T. koningii complex. ITS2 sequences were also useful for locating the biocontrol strains in T. atroviride within the complex T. atroviride-T. koningii. None of the biocontrol strains studied corresponded to biotypes Th2 or Th4 of T. harzianum, which cause mushroom green mold. Correlation between different genotypes and potential biocontrol activity was studied under dual culturing of 17 BCAs in the presence of the phytopathogenic fungi Phoma betae, Rosellinia necatrix, Botrytis cinerea, and Fusarium oxysporum f. sp. dianthi in three different media. PMID:10788356

  2. Effects of Fungal Root Pathogens on the Population Dynamics of Biocontrol Strains of Fluorescent Pseudomonads in the Wheat Rhizosphere

    PubMed Central

    Mazzola, Mark; Cook, R. James

    1991-01-01

    The influences of Gaeumannomyces graminis var. tritici (which causes take-all of wheat), Rhizoctonia solani AG-8 (which causes rhizoctonia root rot of wheat), Pythium irregulare, P. aristosporum, and P. ultimum var. sporangiiferum (which cause pythium root rot of wheat) on the population dynamics of Pseudomonas fluorescens 2-79 and Q72a-80 (bicontrol strains active against take-all and pythium root rot of wheat, respectively) in the wheat rhizosphere were examined. Root infection by either G. graminis var. tritici or R. solani resulted in populations of both bacterial strains that were equal to or significantly larger than their respective populations maintained on roots in the absence of these pathogens. In contrast, the population of strain 2-79 was significantly smaller on roots in the presence of any of the three Pythium species than on noninfected roots and was often below the limits of detection (50 CFU/cm of root) on Pythium-infected roots after 40 days of plant growth. In the presence of either P. aristosporum or P. ultimum var. sporangiiferum, the decline in the population of Q72a-80 was similar to that observed on noninfected roots; however, the population of this strain declined more rapidly on roots infected by P. irregulare than on noninfected roots. Application of metalaxyl (which is selectively inhibitory to Pythium spp.) to soil naturally infestated with Pythium spp. resulted in significantly larger rhizosphere populations of the introduced bacteria over time than on plants grown in the same soil without metalaxyl. It is apparent that root infections by fungal pathogens may either enhance or depress the population of fluorescent pseudomonads introduced for their control, with different strains of pseudomonads reacting differentially to different genera and species of the root pathogens. PMID:16348532

  3. Effect of Meloidogyne incognita inoculum density and application rate of Paecilomyces lilacinus strain 251 on biocontrol efficacy and colonization of egg masses analyzed by real-time quantitative PCR.

    PubMed

    Kiewnick, S; Neumann, S; Sikora, R A; Frey, J E

    2011-01-01

    The fungal biocontrol agent, Paecilomyces lilacinus strain 251 (PL251), was evaluated for its potential to control the root-knot nematode Meloidogyne incognita on tomato at varying application rates and inoculum densities. Conversely to previous studies, significant dose-response relationships could not be established. However, we demonstrated that a preplanting soil treatment with the lowest dose of commercially formulated PL251 (2 × 10(5) CFU/g soil) was already sufficient to reduce root galling by 45% and number of egg masses by 69% when averaged over inoculum densities of 100 to 1,600 eggs and infective juveniles per 100 cm(3) of soil. To determine the role of colonization of M. incognita egg masses by PL251 for biocontrol efficacy, a real-time quantitative polymerase chain reaction (PCR) assay with a detection limit of 10 CFU/egg mass was used. Real-time PCR revealed a significant relationship between egg mass colonization by PL251 and the dose of product applied to soil but no correlation was found between fungal density and biocontrol efficacy or nematode inoculum level. These results demonstrate that rhizosphere competence is not the key mode of action for PL251 in controlling M. incognita on tomato.

  4. Survival of the rhizosphere-competent biocontrol strain Pseudomonas fluorescens NBRI2650 in the soil and phytosphere.

    PubMed

    Nautlyal, C Shekhar; Johri, J K; Singh, H B

    2002-07-01

    Pseudomonas fluorescens NBRI2650 was isolated after screening 360 bacterial strains from the rhizosphere of chickpea (Cicer arietinum L.) grown in fungal-disease-suppressive field soil. The strain was selected because of its high rhizosphere competence and ability to inhibit the growth of Fusarium oxysporum f.sp. ciceri, Rhizoctonia bataticola, and Pythium sp. under in vitro conditions. Survival and colonization of NBRI2650 in the phytosphere of chickpea, cotton (Gossypium hirsutum L.), cucumber (Cucumis sativus L.), and tomato (Lycopersicon seculentum Mill.) were monitored using a chromosomally located rifampicin-marked mutant P. fluorescens NBRI2650R. The strain showed variable ability to invade and survive in the phytosphere of different plants. Chickpea was used as a tester plant for further work, as it was not invaded by NBRI2650R. The interaction between NBRI2650R and F oxysporum fsp. ciceri was studied by both light microscopy and scanning electron microscopy. The lysis of the fungal cell wall by NBRI2650R was clearly demonstrated. Treatment of the chickpea seeds with NBRI2650R in prerelease experiments in the greenhouse using disease-conducive field soils from Jhansi and Kanpur resulted in increased plant growth and did not result in any perturbation of the indigenous microbial community that inhabited the rhizosphere of chickpea compared with nonbacterized seeds. Direct fermentation of diluted NBRI2650R on vermiculite without the need of expensive fermentors offers a reliable process for manufacturing bacterial inoculants in developing countries. Under field conditions, the horizontal and vertical movement of NBRI2650R was restricted to 30 and 60 cm, respectively, and the strain could not survive in the field during the 7 months before the chickpea could be planted for next cropping season. Field trials conducted at Jhansi, Kanpur, and Pantnagar resulted in higher grain yield increase in the bacteria-treated seed compared with the nonbacterized control

  5. Brevibacillus laterosporus strain BPM3, a potential biocontrol agent isolated from a natural hot water spring of Assam, India.

    PubMed

    Saikia, R; Gogoi, D K; Mazumder, S; Yadav, A; Sarma, R K; Bora, T C; Gogoi, B K

    2011-03-20

    A bacterial strain designated as BPM3 isolated from mud of a natural hot water spring of Nambar Wild Life Sanctuary, Assam, India, strongly inhibited growth of phytopathogenic fungi (Fusarium oxysporum f. sp. ciceri, F. semitectum, Magnaporthe grisea and Rhizoctonia oryzae) and gram-positive bacterium (Staphylococcus aureus). The maximum growth and antagonistic activity was recorded at 30°C, pH 8.5 when starch and peptone were amended as carbon and nitrogen sources, respectively. In greenhouse experiment, this bacterium (BPM3) suppressed blast disease of rice by 30-67% and protected the weight loss by 35-56.5%. The maximum disease protection (67%) and weight loss protection (56.5%) were recorded when the bacterium was applied before 2 days of the pathogen inoculation. Antifungal and antibacterial compounds were isolated from the bacterium which also inhibited the growth of these targeted pathogens. The compounds were purified and on spectroscopic analysis of a purified fraction having R(f) 0.22 which showed strong antifungal and antibacterial activity indicated the presence of C-H, carbonyl group, dimethyl group, -CH(2) and methyl group. The bacterium was characterized by morphological, biochemical and molecular approaches and confirmed that the strain BPM3 is Brevibacillus laterosporus.

  6. Draft Genome Sequence of Curtobacterium sp. Strain ER1/6, an Endophytic Strain Isolated from Citrus sinensis with Potential To Be Used as a Biocontrol Agent.

    PubMed

    Garrido, Leandro Maza; Alves, João Marcelo Pereira; Oliveira, Liliane Santana; Gruber, Arthur; Padilla, Gabriel; Araújo, Welington Luiz

    2016-11-17

    Herein, we report a draft genome sequence of the endophytic Curtobacterium sp. strain ER1/6, isolated from a surface-sterilized Citrus sinensis branch, and it presented the capability to control phytopathogens. Functional annotation of the ~3.4-Mb genome revealed 3,100 protein-coding genes, with many products related to known ecological and biotechnological aspects of this bacterium.

  7. Draft Genome Sequence of Curtobacterium sp. Strain ER1/6, an Endophytic Strain Isolated from Citrus sinensis with Potential To Be Used as a Biocontrol Agent

    PubMed Central

    Garrido, Leandro Maza; Alves, João Marcelo Pereira; Oliveira, Liliane Santana; Padilla, Gabriel

    2016-01-01

    Herein, we report a draft genome sequence of the endophytic Curtobacterium sp. strain ER1/6, isolated from a surface-sterilized Citrus sinensis branch, and it presented the capability to control phytopathogens. Functional annotation of the ~3.4-Mb genome revealed 3,100 protein-coding genes, with many products related to known ecological and biotechnological aspects of this bacterium. PMID:27856581

  8. Use of mixed cultures of biocontrol agents to control sheep nematodes.

    PubMed

    Baloyi, M A; Laing, M D; Yobo, K S

    2012-03-23

    Biological control is a promising non-chemical approach for the control of gastrointestinal nematodes of sheep. Use of combinations of biocontrol agents have been reported to be an effective method to increase the efficacy of biological control effects. In this study, combinations of either two Bacillus thuringiensis (Bt) or Clonostachys rosea (C. rosea) isolates and Bt+C. rosea isolates were evaluated in vitro in microtitre plates for their biocontrol activity on sheep nematodes. The Baermann technique was used to extract the surviving L3 larval stages of intestinal nematodes and counted under a dissecting microscope to determine the larval counts. Results indicate that there was a significant reduction of nematode counts due to combination of biocontrol agents (P<0.001). Combinations of Bt isolates reduced nematodes counts by 72.8%, 64% and 29.8%. The results revealed a control level of 57% when C. rosea isolates P3+P8 were combined. Combination of Bt and C. rosea isolates B10+P8 caused the greatest mortality of 76.7%. Most combinations were antagonistic, with only a few combinations showing an additive effect. None were synergistic. The isolate combinations were more effective than when isolates were used alone. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. Evaluation of Clonostachys rosea for control of plant-parasitic nematodes in soil and in roots of carrot and wheat.

    PubMed

    Iqbal, Mudassir; Dubey, Mukesh; McEwan, Kerstin; Menzel, Uwe; Andersson Franko, Mikael; Viketoft, Maria; Funck Jensen, Dan; Karlsson, Magnus

    2017-09-12

    Biological control is a promising approach to reduce plant diseases caused by nematodes. We tested the effect of the fungus Clonostachys rosea strain IK726 inoculation on nematode community composition in a naturally nematode infested soil in a pot experiment, and the effect of C. rosea on plant health. The numbers of plant-parasitic nematode genera extracted from soil and plant roots decreased by 40 to 73 % when C. rosea was applied, while genera of non-parasitic nematodes were not affected. Soil inoculation of C. rosea increased fresh shoot weight and shoot length of wheat plants by 20 and 24 %, respectively, while only shoot dry weight increased by 48 % in carrots. Light microscopy of in vitro C. rosea - nematode interactions did not reveal evidence of direct parasitism. However, culture filtrates of C. rosea growing in potato dextrose broth, malt extract broth and synthetic nutrient broth exhibited toxicity towards nematodes and immobilised 57, 62 and 100 % of the nematodes, respectively, within 48 h. This study demonstrates that C. rosea can control plant-parasitic nematodes and thereby improve plant growth. The most likely mechanism responsible for the antagonism is antibiosis through production of nematicidal compounds, rather than direct parasitism.

  10. Characterization of Two ABC Transporters from Biocontrol and Phytopathogenic Fusarium oxysporus

    USDA-ARS?s Scientific Manuscript database

    ABC transporter genes from four strains of Fusarium oxysporum [two biocontrol and two phytopathogenic (f. sp. lycopersici Race 1) isolates] indicated that this gene is well conserved. However, sequences of promoter regions of FoABC1 differed between 8 phytopathogenic and 11 biocontrol strains of F....

  11. The biocontrol bacterium Pseudomonas fluorescens Pf29Arp strain affects the pathogenesis-related gene expression of the take-all fungus Gaeumannomyces graminis var. tritici on wheat roots.

    PubMed

    Daval, Stéphanie; Lebreton, Lionel; Gazengel, Kévin; Boutin, Morgane; Guillerm-Erckelboudt, Anne-Yvonne; Sarniguet, Alain

    2011-12-01

    The main effects of antagonistic rhizobacteria on plant pathogenic fungi are antibiosis, fungistasis or an indirect constraint through the induction of a plant defence response. To explore different biocontrol mechanisms, an in vitro confrontation assay was conducted with the rhizobacterium Pseudomonas fluorescens Pf29Arp as a biocontrol agent of the fungus Gaeumannomyces graminis var. tritici (Ggt) on wheat roots. In parallel with the assessment of disease extension, together with the bacterial and fungal root colonization rates, the transcript levels of candidate fungal pathogenicity and plant-induced genes were monitored during the 10-day infection process. The bacterial inoculation of wheat roots with the Pf29Arp strain reduced the development of Ggt-induced disease expressed as attack frequency and necrosis length. The growth rates of Ggt and Pf29Arp, monitored through quantitative polymerase chain reaction of DNA amounts with a part of the Ggt 18S rDNA gene and a specific Pf29Arp strain detection probe, respectively, increased throughout the interactions. Bacterial antagonism and colonization had no significant effect on root colonization by Ggt. The expression of fungal and plant genes was quantified in planta by quantitative reverse transcription-polymerase chain reaction during the interactions thanks to the design of specific primers and an innovative universal reference system. During the early stages of the tripartite interaction, several of the fungal genes assayed were down-regulated by Pf29Arp, including two laccases, a β-1,3-exoglucanase and a mitogen-activated protein kinase. The plant host glutathione-S-transferase gene was induced by Ggt alone and up-regulated by Pf29Arp bacteria in interaction with the pathogen. We conclude that Pf29Arp antagonism acts through the alteration of fungal pathogenesis and probably through the activation of host defences.

  12. Efficacy of Eucalyptus cinerea as a Source of Bioactive Compounds for Curative Biocontrol of Crown Gall Caused by Agrobacterium tumefaciens Strain B6.

    PubMed

    Kahla, Yosra; Zouari-Bouassida, Karama; Rezgui, Fatma; Trigui, Mohamed; Tounsi, Slim

    2017-01-01

    This research investigated the Eucalyptus cinerea leaves efficiency in the Agrobacterium tumefaciens biocontrol, the causative agent of crown gall. GC-MS analysis of the essential oil (EO) showed that the main components were 1,8-cineole (61%) and camphene (15.13%). Thanks to its polyphenols, flavonoids, quinones, terpenoids, alkaloids, and tannins richness, the EtOAc-F exhibited the most potent antibacterial activity in vitro. Indeed, compared to the other fractions, it has the lowest MIC and MBC values of 0.312 mg/mL and 2.5 mg/mL, respectively. The GC-MS analysis of EtOAc-F confirmed its richness in antibacterial compounds including gallic acid (7.18%), shikimic acid (5.07%), and catechin (3.12%). The time-kill curve assay of EtOAc-F (2.5 mg/mL) showed a potent bactericidal effect after 20 min of direct contact with A. tumefaciens. In planta experiments, gall weights were significantly reduced when EtOAc-F was applied at 0.625 and 2.5 mg/wounds. Besides, the disease reduction rates in gall weight were 95% and 97.5%, respectively. Interestingly, no phytotoxic effect was observed since tomato seeds germinated in the presence of the different concentrations of EtOAc-F. These results suggest that EtOAc-F has a good potential to be a curative biocontrol agent for crown gall disease.

  13. Partial purification, characterization, and kinetic studies of a low-molecular-weight, alkali-tolerant chitinase enzyme from Bacillus subtilis JN032305, A potential biocontrol strain.

    PubMed

    Shivakumar, Srividya; Karmali, Anika Nayak; Ruhimbana, Charles

    2014-01-01

    A new alkalophilic low-molecular-mass chitinase of 14 kD from the potent biocontrol agent Bacillus subtilis JN032305 was partially purified and enzymology of the chitinase was studied. The enzyme showed optimal pH of 9.0 and temperature of 50°C. The enzyme was found stable during the 60-min incubation at 50 °C. The chitinase was inhibited by group specific agents like IAA, DAN, TLCK, and SDS and metal ions Mg(2+), Ca(2+), Fe(2+), Mn(2+), Ba(2+), and Hg(2+), whereas Zn(2+) did not show significant inhibitory effect against the chitinase. PMSF partially inhibited the enzyme. Substrates specificity tests indicated that the enzyme showed 75% of relative activity on glycol chitin, 58% on carboxymethylcellulose (CMC), 33% on chitin flakes, and 166% laminarin compared to that on colloidal chitin. The enzyme also hydrolyzed 4-methylumbelliferyl-N-acetyl-D-glucosaminide, indicating its chitobiase activity. The chitinase of this study has broad specificity, which could hydrolyze not only the glycosidic bond in GlcNAc-GlcNAc but also that of related carbohydrates with glycosidic linkages. The partially purified chitinase not only showed antifungal activity against Rhizoctonia solani and Colletotrichum gloeosporioides, two potent phytopathogens of chilli, but also increased the germination of chilli seeds when infected with the two potent phytopathogenic fungi.

  14. Biocontrol bacteria selected by a direct plant protection strategy against avocado white root rot show antagonism as a prevalent trait.

    PubMed

    González-Sánchez, M Á; Pérez-Jiménez, R M; Pliego, C; Ramos, C; de Vicente, A; Cazorla, F M

    2010-07-01

    This study was undertaken to study bacterial strains obtained directly for their efficient direct control of the avocado white root rot, thus avoiding prescreening by any other possible mechanism of biocontrol which could bias the selection. A collection of 330 bacterial isolates was obtained from the roots and soil of healthy avocado trees. One hundred and forty-three representative bacterial isolates were tested in an avocado/Rosellinia test system, resulting in 22 presumptive protective strains, all of them identified mainly as Pseudomonas and Bacillus species. These 22 candidate strains were screened in a more accurate biocontrol trial, confirming protection of some strains (4 out of the 22). Analyses of the potential bacterial traits involved in the biocontrol activity suggest that different traits could act jointly in the final biocontrol response, but any of these traits were neither sufficient nor generalized for all the active bacteria. All the protective strains selected were antagonistic against some fungal root pathogens. Diverse bacteria with biocontrol activity could be obtained by a direct plant protection strategy of selection. All the biocontrol strains finally selected in this work were antagonistic, showing that antagonism is a prevalent trait in the biocontrol bacteria selected by a direct plant protection strategy. This is the first report on the isolation of biocontrol bacterial strains using direct plant protection strategy in the system avocado/Rosellinia. Characterization of selected biocontrol bacterial strains obtained by a direct plant protection strategy showed that antagonism is a prevalent trait in the selected strains in this experimental system. This suggests that antagonism could be used as useful strategy to select biocontrol strains. © 2009 The Authors. Journal compilation © 2009 The Society for Applied Microbiology.

  15. Identification of a Novel Small Cysteine-Rich Protein in the Fraction from the Biocontrol Fusarium oxysporum Strain CS-20 that Mitigates Fusarium Wilt Symptoms and Triggers Defense Responses in Tomato

    PubMed Central

    Shcherbakova, Larisa A.; Odintsova, Tatyana I.; Stakheev, Alexander A.; Fravel, Deborah R.; Zavriev, Sergey K.

    2016-01-01

    The biocontrol effect of the non-pathogenic Fusarium oxysporum strain CS-20 against the tomato wilt pathogen F. oxysporum f. sp. lycopersici (FOL) has been previously reported to be primarily plant-mediated. This study shows that CS-20 produces proteins, which elicit defense responses in tomato plants. Three protein-containing fractions were isolated from CS-20 biomass using size exclusion chromatography. Exposure of seedling roots to one of these fractions prior to inoculation with pathogenic FOL strains significantly reduced wilt severity. This fraction initiated an ion exchange response in cultured tomato cells resulting in a reversible alteration of extracellular pH; increased tomato chitinase activity, and induced systemic resistance by enhancing PR-1 expression in tomato leaves. Two other protein fractions were inactive in seedling protection. The main polypeptide (designated CS20EP), which was specifically present in the defense-inducing fraction and was not detected in inactive protein fractions, was identified. The nucleotide sequence encoding this protein was determined, and its complete amino acid sequence was deduced from direct Edman degradation (25 N-terminal amino acid residues) and DNA sequencing. The CS20EP was found to be a small basic cysteine-rich protein with a pI of 9.87 and 23.43% of hydrophobic amino acid residues. BLAST search in the NCBI database showed that the protein is new; however, it displays 48% sequence similarity with a hypothetical protein FGSG_10784 from F. graminearum strain PH-1. The contribution of CS20EP to elicitation of tomato defense responses resulting in wilt mitigating is discussed. PMID:26779237

  16. Comparison of major biocontrol strains of non-aflatoxigenic Aspergillus flavus for the reduction of aflatoxins and cyclopiazonic acid in maize

    USDA-ARS?s Scientific Manuscript database

    Biological control of toxigenic Aspergillus flavus in maize through competitive displacement by non-aflatoxigenic strains was evaluated in a series of field studies. Four sets of experiments were conducted between 2007 to 2009 to assess the competitiveness of non-aflatoxigenic strains when challen...

  17. Biocontrol of Phytophthora Blight and Anthracnose in Pepper by Sequentially Selected Antagonistic Rhizobacteria against Phytophthora capsici.

    PubMed

    Sang, Mee Kyung; Shrestha, Anupama; Kim, Du-Yeon; Park, Kyungseok; Pak, Chun Ho; Kim, Ki Deok

    2013-06-01

    We previously developed a sequential screening procedure to select antagonistic bacterial strains against Phytophthora capsici in pepper plants. In this study, we used a modified screening procedure to select effective biocontrol strains against P. capsici; we evaluated the effect of selected strains on Phytophthora blight and anthracnose occurrence and fruit yield in pepper plants under field and plastic house conditions from 2007 to 2009. We selected four potential biocontrol strains (Pseudomonas otitidis YJR27, P. putida YJR92, Tsukamurella tyrosinosolvens YJR102, and Novosphingobium capsulatum YJR107) among 239 bacterial strains. In the 3-year field tests, all the selected strains significantly (P < 0.05) reduced Phytophthora blight without influencing rhizosphere microbial populations; they showed similar or better levels of disease suppressions than in metalaxyl treatment in the 2007 and 2009 tests, but not in the 2008 test. In the 2-year plastic house tests, all the selected strains significantly (P < 0.05) reduced anthracnose incidence in at least one of the test years, but their biocontrol activities were variable. In addition, strains YJR27, YJR92, and YJR102, in certain harvests, increased pepper fruit numbers in field tests and red fruit weights in plastic house tests. Taken together, these results indicate that the screening procedure is rapid and reliable for the selection of potential biocontrol strains against P. capsici in pepper plants. In addition, these selected strains exhibited biocontrol activities against anthracnose, and some of the strains showed plant growth-promotion activities on pepper fruit.

  18. Biocontrol of Phytophthora Blight and Anthracnose in Pepper by Sequentially Selected Antagonistic Rhizobacteria against Phytophthora capsici

    PubMed Central

    Sang, Mee Kyung; Shrestha, Anupama; Kim, Du-Yeon; Park, Kyungseok; Pak, Chun Ho; Kim, Ki Deok

    2013-01-01

    We previously developed a sequential screening procedure to select antagonistic bacterial strains against Phytophthora capsici in pepper plants. In this study, we used a modified screening procedure to select effective biocontrol strains against P. capsici; we evaluated the effect of selected strains on Phytophthora blight and anthracnose occurrence and fruit yield in pepper plants under field and plastic house conditions from 2007 to 2009. We selected four potential biocontrol strains (Pseudomonas otitidis YJR27, P. putida YJR92, Tsukamurella tyrosinosolvens YJR102, and Novosphingobium capsulatum YJR107) among 239 bacterial strains. In the 3-year field tests, all the selected strains significantly (P < 0.05) reduced Phytophthora blight without influencing rhizosphere microbial populations; they showed similar or better levels of disease suppressions than in metalaxyl treatment in the 2007 and 2009 tests, but not in the 2008 test. In the 2-year plastic house tests, all the selected strains significantly (P < 0.05) reduced anthracnose incidence in at least one of the test years, but their biocontrol activities were variable. In addition, strains YJR27, YJR92, and YJR102, in certain harvests, increased pepper fruit numbers in field tests and red fruit weights in plastic house tests. Taken together, these results indicate that the screening procedure is rapid and reliable for the selection of potential biocontrol strains against P. capsici in pepper plants. In addition, these selected strains exhibited biocontrol activities against anthracnose, and some of the strains showed plant growth-promotion activities on pepper fruit. PMID:25288942

  19. The Systematic Investigation of the Quorum Sensing System of the Biocontrol Strain Pseudomonas chlororaphis subsp. aurantiaca PB-St2 Unveils aurI to Be a Biosynthetic Origin for 3-Oxo-Homoserine Lactones

    PubMed Central

    Bauer, Judith S.; Hauck, Nils; Christof, Lisa; Mehnaz, Samina; Gust, Bertolt; Gross, Harald

    2016-01-01

    The shoot endophytic biocontrol strain Pseudomonas chlororaphis subsp. aurantiaca PB-St2 produces a wide range of exoproducts, including enzymes and antibiotics. The production of exoproducts is commonly tightly regulated. In order to get a deeper insight into the regulatory network of PB-St2, the strain was systematically investigated regarding its quorum sensing systems, both on the genetic and metabolic level. The genome analysis of PB-St2 revealed the presence of four putative acyl homoserine lactone (AHL) biosynthesis genes: phzI, csaI, aurI, and hdtS. LC-MS/MS analyses of the crude supernatant extracts demonstrated that PB-St2 produces eight AHLs. In addition, the concentration of all AHL derivatives was quantified time-resolved in parallel over a period of 42 h during the growth of P. aurantiaca PB-St2, resulting in production curves, which showed differences regarding the maximum levels of the AHLs (14.6 nM– 1.75 μM) and the production period. Cloning and heterologous overexpression of all identified AHL synthase genes in Escherichia coli proved the functionality of the resulting synthases PhzI, CsaI, and AurI. A clear AHL production pattern was assigned to each of these three AHL synthases, while the HdtS synthase did not lead to any AHL production. Furthermore, the heterologous expression study demonstrated unequivocally and for the first time that AurI directs the synthesis of two 3-oxo-AHLs. PMID:27861617

  20. Degradation of the fungal cell wall by clostridial strains isolated from soil subjected to biological soil disinfestation and biocontrol of Fusarium wilt disease of spinach.

    PubMed

    Ueki, Atsuko; Takehara, Toshiaki; Ishioka, Gen; Kaku, Nobuo; Ueki, Katsuji

    2017-10-01

    Biological soil disinfestation (BSD) involves elimination of soil-borne plant pathogens in an environmentally friendly manner. Two anaerobic bacterial strains (H110 and TB8) isolated from BSD-treated soil samples were analyzed for their roles in elimination of pathogenic fungi. The two strains were identified as Clostridium beijerinckii based on 16S rRNA gene sequences and phenotypic properties. The strains fermented various carbohydrates and produced acetate, butyrate, and n-butanol as major products as well as abundant gases (H2 and CO2). For evaluation of the antifungal potential of these strains, cells of a pathogen (spinach wilt disease, Fusarium oxysporum f. sp. spinaciae) were co-inoculated into anaerobic media with each anaerobic strain. After incubation for ~3 weeks at 30 °C, 10-30% of the cells of the pathogen survived when incubated without the anaerobic isolates, whereas the pathogen was eliminated when co-incubated with each anaerobe because of the growth of the anaerobic bacterium. It was found by microscopic examination that mycelial cells of the fungal pathogen were severely degraded during the first 3-7 days of the co-incubation. The two strains utilized major cell wall polysaccharides of ascomycetous fungi-chitosan and ß-1,3-glucan (curdlan and laminarin)-as fermentative substrates added to the medium. Furthermore, both isolates degraded a cell wall preparation isolated from the mycelium of the Fusarium pathogen of spinach wilt disease. We concluded that the two anaerobic strains kill the pathogen of spinach wilt disease by degrading major fungal cell wall components as antifungal activities.

  1. Genomic analysis of the biocontrol strain Pseudomonas fluorescens Pf29Arp with evidence of T3SS and T6SS gene expression on plant roots.

    PubMed

    Marchi, Muriel; Boutin, Morgane; Gazengel, Kévin; Rispe, Claude; Gauthier, Jean-Pierre; Guillerm-Erckelboudt, Anne-Yvonne; Lebreton, Lionel; Barret, Matthieu; Daval, Stéphanie; Sarniguet, Alain

    2013-06-01

    Several bacterial strains of the Pseudomonas genus provide plant growth stimulation, plant protection against pests or bioremediation. Among these bacteria, P. fluorescens Pf29Arp reduces the severity of take-all, a disease caused by the pathogenic fungus Gaeumannomyces graminis var. tritici (Ggt) on wheat roots. In this study, we obtained a draft genome of Pf29Arp and subsequent comparative genomic analyses have revealed that this bacterial strain is closely related to strains of the 'P. brassicacearum-like' subgroup including P. brassicacearum ssp. brassicacearum NFM421 and P. fluorescens F113. Despite an overall chromosomal organization similar to these strains, a number of features including antibiotic synthesis gene clusters from secondary metabolism are not found in the Pf29Arp genome. But Pf29Arp possesses different protein secretion systems including type III (T3SS) and type VI (T6SS) secretion systems. Pf29Arp is the first Pseudomonas sp. strain described with four T6SS clusters (cluster I, II, III and IV). In addition, some protein-coding genes involved in the assembly of these secretion systems are basally expressed during Pf29Arp colonization of healthy wheat roots and display different expression patterns on necrotized roots caused by Ggt. These data suggest a role of T3SS and T6SS in the Pf29Arp adaptation to different root environments.

  2. Transcriptomic profiling to identify genes involved in Fusarium mycotoxin Deoxynivalenol and Zearalenone tolerance in the mycoparasitic fungus Clonostachys rosea.

    PubMed

    Kosawang, Chatchai; Karlsson, Magnus; Jensen, Dan Funck; Dilokpimol, Adiphol; Collinge, David B

    2014-01-22

    Clonostachys rosea strain IK726 is a mycoparasitic fungus capable of controlling mycotoxin-producing Fusarium species, including F. graminearum and F. culmorum, known to produce Zearalenone (ZEA) and Deoxynivalenol (DON). DON is a type B trichothecene known to interfere with protein synthesis in eukaryotes. ZEA is a estrogenic-mimicing mycotoxin that exhibits antifungal growth. C. rosea produces the enzyme zearalenone hydrolase (ZHD101), which degrades ZEA. However, the molecular basis of resistance to DON in C. rosea is not understood. We have exploited a genome-wide transcriptomic approach to identify genes induced by DON and ZEA in order to investigate the molecular basis of mycotoxin resistance C. rosea. We generated DON- and ZEA-induced cDNA libraries based on suppression subtractive hybridization. A total of 443 and 446 sequenced clones (corresponding to 58 and 65 genes) from the DON- and ZEA-induced library, respectively, were analysed. DON-induced transcripts represented genes encoding metabolic enzymes such as cytochrome P450, cytochrome c oxidase and stress response proteins. In contrast, transcripts encoding the ZEA-detoxifying enzyme ZHD101 and those encoding a number of ATP-Binding Cassette (ABC) transporter transcripts were highly frequent in the ZEA-induced library. Subsequent bioinformatics analysis predicted that all transcripts with similarity to ABC transporters could be ascribed to only 2 ABC transporters genes, and phylogenetic analysis of the predicted ABC transporters suggested that they belong to group G (pleiotropic drug transporters) of the fungal ABC transporter gene family. This is the first report suggesting involvement of ABC transporters in ZEA tolerance. Expression patterns of a selected set of DON- and ZEA-induced genes were validated by the use of quantitative RT-PCR after exposure to the toxins. The qRT-PCR results obtained confirm the expression patterns suggested from the EST redundancy data. The present study identifies a

  3. Transcriptomic profiling to identify genes involved in Fusarium mycotoxin Deoxynivalenol and Zearalenone tolerance in the mycoparasitic fungus Clonostachys rosea

    PubMed Central

    2014-01-01

    Background Clonostachys rosea strain IK726 is a mycoparasitic fungus capable of controlling mycotoxin-producing Fusarium species, including F. graminearum and F. culmorum, known to produce Zearalenone (ZEA) and Deoxynivalenol (DON). DON is a type B trichothecene known to interfere with protein synthesis in eukaryotes. ZEA is a estrogenic-mimicing mycotoxin that exhibits antifungal growth. C. rosea produces the enzyme zearalenone hydrolase (ZHD101), which degrades ZEA. However, the molecular basis of resistance to DON in C. rosea is not understood. We have exploited a genome-wide transcriptomic approach to identify genes induced by DON and ZEA in order to investigate the molecular basis of mycotoxin resistance C. rosea. Results We generated DON- and ZEA-induced cDNA libraries based on suppression subtractive hybridization. A total of 443 and 446 sequenced clones (corresponding to 58 and 65 genes) from the DON- and ZEA-induced library, respectively, were analysed. DON-induced transcripts represented genes encoding metabolic enzymes such as cytochrome P450, cytochrome c oxidase and stress response proteins. In contrast, transcripts encoding the ZEA-detoxifying enzyme ZHD101 and those encoding a number of ATP-Binding Cassette (ABC) transporter transcripts were highly frequent in the ZEA-induced library. Subsequent bioinformatics analysis predicted that all transcripts with similarity to ABC transporters could be ascribed to only 2 ABC transporters genes, and phylogenetic analysis of the predicted ABC transporters suggested that they belong to group G (pleiotropic drug transporters) of the fungal ABC transporter gene family. This is the first report suggesting involvement of ABC transporters in ZEA tolerance. Expression patterns of a selected set of DON- and ZEA-induced genes were validated by the use of quantitative RT-PCR after exposure to the toxins. The qRT-PCR results obtained confirm the expression patterns suggested from the EST redundancy data. Conclusion The

  4. Draft Genome Sequence of Stenotrophomonas maltophilia Strain B418, a Promising Agent for Biocontrol of Plant Pathogens and Root-Knot Nematode.

    PubMed

    Wu, Yuanzheng; Wang, Yilian; Li, Jishun; Hu, Jindong; Chen, Kai; Wei, Yanli; Bazhanov, Dmitry P; Bazhanova, Alesia A; Yang, Hetong

    2015-02-19

    Stenotrophomonas maltophilia strain B418 was isolated from a barley rhizosphere in China. This bacterium exhibits broad-spectrum inhibitory activities against plant pathogens and root-knot nematode along with growth-promoting effects. Here, we present the draft genome sequence of S. maltophilia B418.

  5. Expression of a Serine Protease Gene prC Is Up-Regulated by Oxidative Stress in the Fungus Clonostachys rosea: Implications for Fungal Survival

    PubMed Central

    Liu, Wen-Jing; Zhou, Wei; Tao, Nan; Tu, Hui-Hui; Huang, Xiao-Wei; Yang, Jin-Kui; Zhang, Ke-Qin

    2010-01-01

    Background Soil fungi face a variety of environmental stresses such as UV light, high temperature, and heavy metals. Adaptation of gene expression through transcriptional regulation is a key mechanism in fungal response to environmental stress. In Saccharomyces cerevisiae, the transcription factors Msn2/4 induce stress-mediated gene expression by binding to the stress response element. Previous studies have demonstrated that the expression of extracellular proteases is up-regulated in response to heat shock in fungi. However, the physiological significance of regulation of these extracellular proteases by heat shock remains unclear. The nematophagous fungus Clonostachys rosea can secret an extracellular serine protease PrC during the infection of nematodes. Since the promoter of prC has three copies of the stress response element, we investigated the effect of environmental stress on the expression of prC. Methodology/Principal Findings Our results demonstrated that the expression of prC was up-regulated by oxidants (H2O2 or menadione) and heat shock, most likely through the stress response element. After oxidant treatment or heat shock, the germination of conidia in the wild type strain was significantly higher than that in the prC mutant strain in the presence of nematode cuticle. Interestingly, the addition of nematode cuticle significantly attenuated the production of reactive oxygen species (ROS) induced by oxidants and heat shock in the wild type strain, but not in prC mutant strain. Moreover, low molecule weight (<3 kD) degradation products of nematode cuticle suppressed the inhibitory effect of conidial germination induced by oxidants and heat shock. Conclusions/Significance These results indicate that PrC plays a protective role in oxidative stress in C. rosea. PrC degrades the nematode cuticle to produce degradation products, which in turn offer a protective effect against oxidative stress by scavenging ROS. Our study reveals a novel strategy for fungi to

  6. Biocontrol and plant growth promoting activities of a Streptomyces corchorusii strain UCR3-16 and preparation of powder formulation for application as biofertilizer agents for rice plant.

    PubMed

    Tamreihao, K; Ningthoujam, Debananda S; Nimaichand, Salam; Singh, Elangbam Shanta; Reena, Pascal; Singh, Salam Herojeet; Nongthomba, Upendra

    2016-11-01

    Streptomyces corchorusii strain UCR3-16, obtained from rice rhizospheric soils showed antifungal activities against 6 major rice fungal pathogens by diffusible and volatile compounds production. The strain was found positive for production of fungal cell wall degrading enzymes such as chitinase, β-1,3-glucanase, β-1,4-glucanase, lipase and protease. The strain was also positive for plant growth promoting traits. It produced up to 30.5μg/ml of IAA and solubilized a significant amount of inorganic phosphate (up to 102μg/ml). It also produced 69% siderophore units. The strain also produced ammonia and gave positive result for ACC deaminase activity. Highest vigor index of inoculated seedlings was observed when rice seeds were treated with cell suspension of UCR3-16 corresponding to 4.5×10(8)cfu/ml. Bioinoculant-treated seeds also showed similar results under pathogen challenged conditions. In pot trial experiments, UCR3-16-treated rice plants showed significantly increased growth and grain yield production. Powder formulation of the strain was developed using talcum and corn starch as carriers and the shelf-lives were monitored. Talcum formulation showed higher cell-count than corn starch even after 6 months of storage, and optimum condition for storage of the powder formulation were found to be at 4°C. Pot trial experiments using talcum powder formulation also showed significant positive effects on growth of rice plants. Field trial using talcum powder formulation also exhibited significant enhancement in shoot length and weight of shoot and root, and total grain yield and weight of grains in rice plants. Talcum formulation also significantly reduced the sheath blight disease in rice leaves. Copyright © 2016. Published by Elsevier GmbH.

  7. Posttranscriptional Repression of GacS/GacA-Controlled Genes by the RNA-Binding Protein RsmE Acting Together with RsmA in the Biocontrol Strain Pseudomonas fluorescens CHA0

    PubMed Central

    Reimmann, Cornelia; Valverde, Claudio; Kay, Elisabeth; Haas, Dieter

    2005-01-01

    In the plant-beneficial soil bacterium Pseudomonas fluorescens CHA0, the production of biocontrol factors (antifungal secondary metabolites and exoenzymes) is controlled at a posttranscriptional level by the GacS/GacA signal transduction pathway involving RNA-binding protein RsmA as a key regulatory element. This protein is assumed to bind to the ribosome-binding site of target mRNAs and to block their translation. RsmA-mediated repression is relieved at the end of exponential growth by two GacS/GacA-controlled regulatory RNAs RsmY and RsmZ, which bind and sequester the RsmA protein. A gene (rsmE) encoding a 64-amino-acid RsmA homolog was identified and characterized in strain CHA0. Overexpression of rsmE strongly reduced the expression of target genes (hcnA, for a hydrogen cyanide synthase subunit; aprA, for the main exoprotease; and phlA, for a component of 2,4-diacetylphloroglucinol biosynthesis). Single null mutations in either rsmA or rsmE resulted in a slight increase in the expression of hcnA, aprA, and phlA. By contrast, an rsmA rsmE double mutation led to strongly increased and advanced expression of these target genes and completely suppressed a gacS mutation. Both the RsmE and RsmA levels increased with increasing cell population densities in strain CHA0; however, the amount of RsmA showed less variability during growth. Expression of rsmE was controlled positively by GacA and negatively by RsmA and RsmE. Mobility shift assays demonstrated specific binding of RsmE to RsmY and RsmZ RNAs. The transcription and stability of both regulatory RNAs were strongly reduced in the rsmA rsmE double mutant. In conclusion, RsmA and RsmE together account for maximal repression in the GacS/GacA cascade of strain CHA0. PMID:15601712

  8. The use of newly isolated Streptomyces strain TN258 as potential biocontrol agent of potato tubers leak caused by Pythium ultimum.

    PubMed

    Sellem, Imen; Triki, Mohamed Ali; Elleuch, Lobna; Cheffi, Manel; Chakchouk, Ahlem; Smaoui, Slim; Mellouli, Lotfi

    2017-05-01

    An actinomycete strain designated TN258, was isolated from Tunisian Sahara soil and selected for its antagonistic activity, especially against Pythium ultimum (P. ultimum) causing potato tubers leak. Based on the results of cultural characteristic of TN258 strain, the 16S rRNA gene nucleotide sequence (1433 bp, accession n° HE600071) and the phylogenetic analysis, we propose the assignment of our new isolate bacterium as Streptomyces TN258 strain. After culture optimization, the inhibitory effect of TN258 free cell supernatant against P. ultimum was evaluated. As result, by application of 50% (v/v) from 25 mg ml(-1) of concentration, mycelial growth was totally inhibited with hyphal destruction. At the same concentration, the oospores were distorted and the germination was completely stopped. In potato tubers, Streptomyces TN258 filtrated supernatant, applied 24 h before inoculation by P. ultimum (preventive treatment group) was able to significantly decrease pathogen penetration by 62% and to reduce the percentage of weight loss by 59.43%, in comparison with non-treated group. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Conjunctively screening of biocontrol agents (BCAs) against fusarium root rot and fusarium head blight caused by Fusarium graminearum.

    PubMed

    Wang, Lu-Yao; Xie, Yue-Shen; Cui, Yuan-Yu; Xu, Jianjun; He, Wei; Chen, Huai-Gu; Guo, Jian-Hua

    2015-08-01

    Fusarium root-rot and fusarium head blight are plant diseases caused by Fusarium sp. in different growth periods of wheat, bring heavy losses to crop production in China. This research is aiming to screen biocontrol agents conjunctively for controlling these two diseases at the same time, as well as evaluate our previous BCAs (Biological Control Agents) screening strategies in more complex situation, considering biocontrol is well concerned as an environmental-friendly plant disease controlling method. Totally 966 bacterial isolates were screened from different parts of wheat tissues, of which potential biocontrol values were detected according to their abilities in antagonism inhibition and secreting extracellular hydrolytic enzyme. Biocontrol tests against fusarium root rot and fusarium head blight were carried out on 37 bacterial isolates with potential biocontrol capacity after pre-selection through ARDRA- and BOX-PCR analysis on strains with high assessment points. We acquired 10 BCAs with obvious biocontrol efficacy (more than 40%) in greenhouse and field tests. Pseudomonas fluorescens LY1-8 performed well in both two tests (biocontrol efficacy: 44.62% and 58.31%), respectively. Overall, correlation coefficient is 0.720 between assessment values of 37 tested BCA strains and their biocontrol efficacy in trails against fusarium root rot; correlation coefficient is 0.806 between their assessment values and biocontrol efficacy in trails against fusarium head blight. We acquired 10 well-performed potential BCAs, especially P. fluorescens LY1-8 displayed good biocontrol capacity against two different diseases on wheat. Biocontrol efficacies results in both greenhouse and field tests showed high positive correlation with assessment values (0.720 and 0.806), suggesting that the BCAs screening and assessing strategy previously developed in our lab is also adaptable for conjunctively screening BCAs for controlling both root and shoot diseases on wheat caused by same

  10. Clonostachys rosea demethiolase STR3 controls the conversion of methionine into methanethiol

    PubMed Central

    Jia, Kai-Zhi; Zhang, Quan; Sun, Lin-Yang; Xu, Yang-Hua; Li, Hong-Mei; Tang, Ya-Jie

    2016-01-01

    Eukaryote-derived methioninase, catalyzing the one-step degradation of methionine (Met) to methanethiol (MTL), has received much attention for its low immunogenic potential and use as a therapeutic agent against Met-dependent tumors. Although biological and chemical degradation pathways for Met-MTL conversion are proposed, the concrete molecular mechanism for Met-MTL conversion in eukaryotes is still unclear. Previous studies demonstrated that α-keto-methylthiobutyric acid (KMBA), the intermediate for Met-MTL conversion, was located extracellularly and the demethiolase STR3 possessed no activities towards Met, which rule out the possibility of intracellular Met-MTL conversion pathway inside eukaryotes. We report here that degradation of Met resulted in intracellular accumulation of KMBA in Clonostachys rosea. Addition of Met to culture media led to the production of MTL and downregulation of STR3, while incubation of Met with surrogate substrate α-ketoglutaric acid enhanced the synthesis of MTL and triggered the upregulation of STR3. Subsequent biochemical analysis with recombinant STR3 showed that STR3 directly converted both Met and its transamination product KMBA to MTL. These results indicated that STR3 as rate-limiting enzyme degrades Met and KMBA into MTL. Our findings suggest STR3 is a potential target for therapeutic agents against Met-dependent tumors and aging. PMID:26902928

  11. Clonostachys rosea demethiolase STR3 controls the conversion of methionine into methanethiol.

    PubMed

    Jia, Kai-Zhi; Zhang, Quan; Sun, Lin-Yang; Xu, Yang-Hua; Li, Hong-Mei; Tang, Ya-Jie

    2016-02-23

    Eukaryote-derived methioninase, catalyzing the one-step degradation of methionine (Met) to methanethiol (MTL), has received much attention for its low immunogenic potential and use as a therapeutic agent against Met-dependent tumors. Although biological and chemical degradation pathways for Met-MTL conversion are proposed, the concrete molecular mechanism for Met-MTL conversion in eukaryotes is still unclear. Previous studies demonstrated that α-keto-methylthiobutyric acid (KMBA), the intermediate for Met-MTL conversion, was located extracellularly and the demethiolase STR3 possessed no activities towards Met, which rule out the possibility of intracellular Met-MTL conversion pathway inside eukaryotes. We report here that degradation of Met resulted in intracellular accumulation of KMBA in Clonostachys rosea. Addition of Met to culture media led to the production of MTL and downregulation of STR3, while incubation of Met with surrogate substrate α-ketoglutaric acid enhanced the synthesis of MTL and triggered the upregulation of STR3. Subsequent biochemical analysis with recombinant STR3 showed that STR3 directly converted both Met and its transamination product KMBA to MTL. These results indicated that STR3 as rate-limiting enzyme degrades Met and KMBA into MTL. Our findings suggest STR3 is a potential target for therapeutic agents against Met-dependent tumors and aging.

  12. Potential biocontrol actinobacteria: Rhizospheric isolates from the Argentine Pampas lowlands legumes.

    PubMed

    Solans, Mariana; Scervino, Jose Martin; Messuti, María Inés; Vobis, Gernot; Wall, Luis Gabriel

    2016-11-01

    Control of fungal plant diseases by using naturally occurring non-pathogenic microorganisms represents a promising approach to biocontrol agents. This study reports the isolation, characterization, and fungal antagonistic activity of actinobacteria from forage soils in the Flooding Pampa, Argentina. A total of 32 saprophytic strains of actinobacteria were obtained by different isolation methods from rhizospheric soil of Lotus tenuis growing in the Salado River Basin. Based on physiological traits, eight isolates were selected for their biocontrol-related activities such as production of lytic extracellular enzymes, siderophores, hydrogen cyanide (HCN), and antagonistic activity against Cercospora sojina, Macrophomia phaseolina, Phomopsis sp., Fusarium oxysporum, and Fusarium verticilloides. These actinobacteria strains were characterized morphologically, physiologically, and identified by using molecular techniques. The characterization of biocontrol-related activities in vitro showed positive results for exoprotease, phospholipase, fungal growth inhibition, and siderophore production. However, none of the strains was positive for the production of hydrogen cyanide (HCN). Streptomyces sp. MM140 presented the highest index for biocontrol, and appear to be promising pathogenic fungi biocontrol agents. These results show the potential capacity of actinobacteria isolated from forage soils in the Argentine Pampas lowlands as promising biocontrol agents, and their future agronomic applications.

  13. Analysis of the pmsCEAB gene cluster involved in biosynthesis of salicylic acid and the siderophore pseudomonine in the biocontrol strain Pseudomonas fluorescens WCS374.

    PubMed

    Mercado-Blanco, J; van der Drift, K M; Olsson, P E; Thomas-Oates, J E; van Loon, L C; Bakker, P A

    2001-03-01

    Mutants of Pseudomonas fluorescens WCS374 defective in biosynthesis of the fluorescent siderophore pseudobactin still display siderophore activity, indicating the production of a second siderophore. A recombinant cosmid clone (pMB374-07) of a WCS374 gene library harboring loci necessary for the biosynthesis of salicylic acid (SA) and this second siderophore pseudomonine was isolated. The salicylate biosynthesis region of WCS374 was localized in a 5-kb EcoRI fragment of pMB374-07. The SA and pseudomonine biosynthesis region was identified by transfer of cosmid pMB374-07 to a pseudobactin-deficient strain of P. putida. Sequence analysis of the 5-kb subclone revealed the presence of four open reading frames (ORFs). Products of two ORFs (pmsC and pmsB) showed homologies with chorismate-utilizing enzymes; a third ORF (pmsE) encoded a protein with strong similarity with enzymes involved in the biosynthesis of siderophores in other bacterial species. The region also contained a putative histidine decarboxylase gene (pmsA). A putative promoter region and two predicted iron boxes were localized upstream of pmsC. We determined by reverse transcriptase-mediated PCR that the pmsCEAB genes are cotranscribed and that expression is iron regulated. In vivo expression of SA genes was achieved in P. putida and Escherichia coli cells. In E. coli, deletions affecting the first ORF (pmsC) diminished SA production, whereas deletion of pmsB abolished it completely. The pmsB gene induced low levels of SA production in E. coli when expressed under control of the lacZ promoter. Several lines of evidence indicate that SA and pseudomonine biosynthesis are related. Moreover, we isolated a Tn5 mutant (374-05) that is simultaneously impaired in SA and pseudomonine production.

  14. Analysis of the pmsCEAB Gene Cluster Involved in Biosynthesis of Salicylic Acid and the Siderophore Pseudomonine in the Biocontrol Strain Pseudomonas fluorescens WCS374

    PubMed Central

    Mercado-Blanco, Jesús; van der Drift, Koen M. G. M.; Olsson, Per E.; Thomas-Oates, Jane E.; van Loon, Leendert C.; Bakker, Peter A. H. M.

    2001-01-01

    Mutants of Pseudomonas fluorescens WCS374 defective in biosynthesis of the fluorescent siderophore pseudobactin still display siderophore activity, indicating the production of a second siderophore. A recombinant cosmid clone (pMB374-07) of a WCS374 gene library harboring loci necessary for the biosynthesis of salicylic acid (SA) and this second siderophore pseudomonine was isolated. The salicylate biosynthesis region of WCS374 was localized in a 5-kb EcoRI fragment of pMB374-07. The SA and pseudomonine biosynthesis region was identified by transfer of cosmid pMB374-07 to a pseudobactin-deficient strain of P. putida. Sequence analysis of the 5-kb subclone revealed the presence of four open reading frames (ORFs). Products of two ORFs (pmsC and pmsB) showed homologies with chorismate-utilizing enzymes; a third ORF (pmsE) encoded a protein with strong similarity with enzymes involved in the biosynthesis of siderophores in other bacterial species. The region also contained a putative histidine decarboxylase gene (pmsA). A putative promoter region and two predicted iron boxes were localized upstream of pmsC. We determined by reverse transcriptase-mediated PCR that the pmsCEAB genes are cotranscribed and that expression is iron regulated. In vivo expression of SA genes was achieved in P. putida and Escherichia coli cells. In E. coli, deletions affecting the first ORF (pmsC) diminished SA production, whereas deletion of pmsB abolished it completely. The pmsB gene induced low levels of SA production in E. coli when expressed under control of the lacZ promoter. Several lines of evidence indicate that SA and pseudomonine biosynthesis are related. Moreover, we isolated a Tn5 mutant (374-05) that is simultaneously impaired in SA and pseudomonine production. PMID:11222588

  15. Pseudomonas fluorescens F113 Mutant with Enhanced Competitive Colonization Ability and Improved Biocontrol Activity against Fungal Root Pathogens ▿

    PubMed Central

    Barahona, Emma; Navazo, Ana; Martínez-Granero, Francisco; Zea-Bonilla, Teresa; Pérez-Jiménez, Rosa María; Martín, Marta; Rivilla, Rafael

    2011-01-01

    Motility is one of the most important traits for efficient rhizosphere colonization by Pseudomonas fluorescens F113rif (F113). In this bacterium, motility is a polygenic trait that is repressed by at least three independent pathways, including the Gac posttranscriptional system, the Wsp chemotaxis-like pathway, and the SadB pathway. Here we show that the kinB gene, which encodes a signal transduction protein that together with AlgB has been implicated in alginate production, participates in swimming motility repression through the Gac pathway, acting downstream of the GacAS two-component system. Gac mutants are impaired in secondary metabolite production and are unsuitable as biocontrol agents. However, the kinB mutant and a triple mutant affected in kinB, sadB, and wspR (KSW) possess a wild-type phenotype for secondary metabolism. The KSW strain is hypermotile and more competitive for rhizosphere colonization than the wild-type strain. We have compared the biocontrol activity of KSW with those of the wild-type strain and a phenotypic variant (F113v35 [V35]) which is hypermotile and hypercompetitive but is affected in secondary metabolism since it harbors a gacS mutation. Biocontrol experiments in the Fusarium oxysporum f. sp. radicis-lycopersici/Lycopersicum esculentum (tomato) and Phytophthora cactorum/Fragaria vesca (strawberry) pathosystems have shown that the three strains possess biocontrol activity. Biocontrol activity was consistently lower for V35, indicating that the production of secondary metabolites was the most important trait for biocontrol. Strain KSW showed improved biocontrol compared with the wild-type strain, indicating that an increase in competitive colonization ability resulted in improved biocontrol and that the rational design of biocontrol agents by mutation is feasible. PMID:21685161

  16. Pseudomonas fluorescens F113 mutant with enhanced competitive colonization ability and improved biocontrol activity against fungal root pathogens.

    PubMed

    Barahona, Emma; Navazo, Ana; Martínez-Granero, Francisco; Zea-Bonilla, Teresa; Pérez-Jiménez, Rosa María; Martín, Marta; Rivilla, Rafael

    2011-08-01

    Motility is one of the most important traits for efficient rhizosphere colonization by Pseudomonas fluorescens F113rif (F113). In this bacterium, motility is a polygenic trait that is repressed by at least three independent pathways, including the Gac posttranscriptional system, the Wsp chemotaxis-like pathway, and the SadB pathway. Here we show that the kinB gene, which encodes a signal transduction protein that together with AlgB has been implicated in alginate production, participates in swimming motility repression through the Gac pathway, acting downstream of the GacAS two-component system. Gac mutants are impaired in secondary metabolite production and are unsuitable as biocontrol agents. However, the kinB mutant and a triple mutant affected in kinB, sadB, and wspR (KSW) possess a wild-type phenotype for secondary metabolism. The KSW strain is hypermotile and more competitive for rhizosphere colonization than the wild-type strain. We have compared the biocontrol activity of KSW with those of the wild-type strain and a phenotypic variant (F113v35 [V35]) which is hypermotile and hypercompetitive but is affected in secondary metabolism since it harbors a gacS mutation. Biocontrol experiments in the Fusarium oxysporum f. sp. radicis-lycopersici/Lycopersicum esculentum (tomato) and Phytophthora cactorum/Fragaria vesca (strawberry) pathosystems have shown that the three strains possess biocontrol activity. Biocontrol activity was consistently lower for V35, indicating that the production of secondary metabolites was the most important trait for biocontrol. Strain KSW showed improved biocontrol compared with the wild-type strain, indicating that an increase in competitive colonization ability resulted in improved biocontrol and that the rational design of biocontrol agents by mutation is feasible.

  17. An In vitro Study of Bio-Control and Plant Growth Promotion Potential of Salicaceae Endophytes.

    PubMed

    Kandel, Shyam L; Firrincieli, Andrea; Joubert, Pierre M; Okubara, Patricia A; Leston, Natalie D; McGeorge, Kendra M; Mugnozza, Giuseppe S; Harfouche, Antoine; Kim, Soo-Hyung; Doty, Sharon L

    2017-01-01

    Microbial communities in the endosphere of Salicaceae plants, poplar (Populus trichocarpa) and willow (Salix sitchensis), have been demonstrated to be important for plant growth promotion, protection from biotic and abiotic stresses, and degradation of toxic compounds. Our study aimed to investigate bio-control activities of Salicaceae endophytes against various soil borne plant pathogens including Rhizoctonia solani AG-8, Fusarium culmorum, Gaeumannomyces graminis var. tritici, and Pythium ultimum. Additionally, different plant growth promoting traits such as biological nitrogen fixation (BNF), indole-3-acetic acid (IAA) biosynthesis, phosphate solubilization, and siderophore production were assessed in all bio-control positive strains. Burkholderia, Rahnella, Pseudomonas, and Curtobacterium were major endophyte genera that showed bio-control activities in the in-vitro assays. The bio-control activities of Burkholderia strains were stronger across all tested plant pathogens as compared to other stains. Genomes of sequenced Burkholderia strains WP40 and WP42 were surveyed to identify the putative genes involved in the bio-control activities. The ocf and hcnABC gene clusters responsible for biosynthesis of the anti-fungal metabolites, occidiofungin and hydrogen cyanide, are present in the genomes of WP40 and WP42. Nearly all endophyte strains showing the bio-control activities produced IAA, solubilized tricalcium phosphate, and synthesized siderophores in the culture medium. Moreover, some strains reduced acetylene into ethylene in the acetylene reduction assay, a common assay used for BNF. Salicaceae endophytes could be useful for bio-control of various plant pathogens, and plant growth promotion possibly through the mechanisms of BNF, IAA production, and nutrient acquisition.

  18. An In vitro Study of Bio-Control and Plant Growth Promotion Potential of Salicaceae Endophytes

    PubMed Central

    Kandel, Shyam L.; Firrincieli, Andrea; Joubert, Pierre M.; Okubara, Patricia A.; Leston, Natalie D.; McGeorge, Kendra M.; Mugnozza, Giuseppe S.; Harfouche, Antoine; Kim, Soo-Hyung; Doty, Sharon L.

    2017-01-01

    Microbial communities in the endosphere of Salicaceae plants, poplar (Populus trichocarpa) and willow (Salix sitchensis), have been demonstrated to be important for plant growth promotion, protection from biotic and abiotic stresses, and degradation of toxic compounds. Our study aimed to investigate bio-control activities of Salicaceae endophytes against various soil borne plant pathogens including Rhizoctonia solani AG-8, Fusarium culmorum, Gaeumannomyces graminis var. tritici, and Pythium ultimum. Additionally, different plant growth promoting traits such as biological nitrogen fixation (BNF), indole-3-acetic acid (IAA) biosynthesis, phosphate solubilization, and siderophore production were assessed in all bio-control positive strains. Burkholderia, Rahnella, Pseudomonas, and Curtobacterium were major endophyte genera that showed bio-control activities in the in-vitro assays. The bio-control activities of Burkholderia strains were stronger across all tested plant pathogens as compared to other stains. Genomes of sequenced Burkholderia strains WP40 and WP42 were surveyed to identify the putative genes involved in the bio-control activities. The ocf and hcnABC gene clusters responsible for biosynthesis of the anti-fungal metabolites, occidiofungin and hydrogen cyanide, are present in the genomes of WP40 and WP42. Nearly all endophyte strains showing the bio-control activities produced IAA, solubilized tricalcium phosphate, and synthesized siderophores in the culture medium. Moreover, some strains reduced acetylene into ethylene in the acetylene reduction assay, a common assay used for BNF. Salicaceae endophytes could be useful for bio-control of various plant pathogens, and plant growth promotion possibly through the mechanisms of BNF, IAA production, and nutrient acquisition. PMID:28348550

  19. Environmental factors affect the activity of biocontrol agents against ochratoxigenic Aspergillus carbonarius on wine grape.

    PubMed

    De Curtis, F; de Felice, D V; Ianiri, G; De Cicco, V; Castoria, R

    2012-09-17

    The influence of temperature and relative humidity (RH) on the activity of three biocontrol agents-the yeast Metschnikowia pulcherrima LS16 and two strains of the yeast-like fungus Aureobasidium pullulans LS30 and AU34-2-against infection by A. carbonarius and ochratoxin A (OTA) accumulation in wine grape berries was investigated in lab-scale experiments. The presence of wounds on grape skin dramatically favored infection of berries by A. carbonarius strain A1102, since unwounded berries showed very low levels of infection at all conditions of RH and temperature tested. Artificially wounded berries pre-treated with the biocontrol agents were inoculated with the ochratoxigenic A. carbonarius strain A1102 and were incubated for 5 days at two levels of RH (60% and 100%) and three different temperatures (20, 25 and 30 °C). The three biocontrol agents were able to prevent infections at 60% RH and 20 °C. At 60% RH and 25 °C only strain AU34-2 achieved some protection on day 5, whereas at 30 °C a limited biocontrol efficacy was evident only up to day 2. At 100% RH, LS16, LS30 and AU34-2 showed effective protection of grape berries at 20 °C until the 5th day of incubation. The three biocontrol agents achieved significant protection at higher temperatures only until the 2nd day after the beginning of the experiment: all three strains at 25 °C, and only strain LS16 at 30 °C. After 5 days, the three biocontrol agents were able to significantly reduce the level of OTA in berries at all the conditions tested. This occurred even when protection from infection was not significant, except at 30 °C and 100% of RH for all the three strains, and at 25 °C and 100% of RH for strain LS16. The biocontrol agents displayed a higher rate of colonization on grape berries at 20 and 25 °C than at 30 °C. The higher value of RH (100%) appeared to increase the rate of colonization, in particular at 20 and 25 °C. Taken together, our results emphasize the significant influence of

  20. Predacious bacteria, Bdellovibrio with potential for biocontrol.

    PubMed

    Markelova, Natalia Y

    2010-11-01

    Bacteria of the genus of Bdellovibrio are highly motile Gram-negative predators of other Gram-negative bacteria causing lysis of their prey. Here we report results of studies on the interactions of Bdellovibrio with species of Alcaligenes, Campylobacter, Erwinia, Escherichia, Helicobacter, Pseudomonas, Legionella, and Shigella in agar lower, liquid media and cells attached to a surface. Helicobacter pylori was studied employing both actively growing and viable but nonculturable (VBNC) cells. The majority of the bacterial strains tested were found to be susceptible to Bdellovibrio. A significant observation was that Bdellovibrio attacked both actively growing and VBNC H. pylori, that phenomenon has never been reported. The results indicate that bdellovibrios have potential as biocontrol agents.

  1. Complete Genome Sequence of Ralstonia solanacearum FJAT-1458, a Potential Biocontrol Agent for Tomato Wilt

    PubMed Central

    Chen, Deju; Zhu, Yujing; Wang, Jieping; Chen, Zheng; Che, Jiamei; Zheng, Xuefang; Chen, Xiaoqiang

    2017-01-01

    ABSTRACT An avirulent strain of Ralstonia solanacearum FJAT-1458 was isolated from a living tomato. Here, we report the complete R. solanacearum FJAT-1458 genome sequence of 6,059,899 bp and 5,241 genes. This bacterial strain is a potential candidate as a biocontrol agent in the form of a plant vaccine for bacterial wilt. PMID:28385834

  2. Surfactin triggers biofilm formation of Bacillus subtilis in melon phylloplane and contributes to the biocontrol activity.

    PubMed

    Zeriouh, Houda; de Vicente, Antonio; Pérez-García, Alejandro; Romero, Diego

    2014-07-01

    The biocontrol activity of many Bacillus species has been traditionally related to the direct antagonism of pathogens. In previous works, we reported that B. subtilis strain UMAF6614 was an efficient biocontrol agent that produced bacillomycin, fengycin and surfactin lipopeptides. Bacillomycins and fengycins were shown to have antagonistic activity towards fungal and bacterial pathogens of cucurbits; however, the functionality of surfactin remained unclear. In this study, the role of surfactin in the biocontrol activity of this strain was investigated. We observed that a deficiency in surfactin production led to a partial reduction of disease suppression by this biocontrol agent, which coincided with a defect in biofilm formation and the colonization of the melon phylloplane. These effects were due to a dramatic reduction in the production of exopolysaccharide and the TasA protein, which are the two major components of the extracellular matrix. We propose that the biocontrol activity of this strain is the result of the coordinated action of the three families of lipopeptides. B. subtilis UMAF6614 produces surfactin to trigger biofilm formation on melon phylloplane, which ensures the long-term persistence and the adequate secretion of suppressive lipopeptides, bacillomycins and fengycins, which efficiently target pathogens. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

  3. Whole genome shotgun sequence of Bacillus amyloliquefaciens TF28, a biocontrol entophytic bacterium.

    PubMed

    Zhang, Shumei; Jiang, Wei; Li, Jing; Meng, Liqiang; Cao, Xu; Hu, Jihua; Liu, Yushuai; Chen, Jingyu; Sha, Changqing

    2016-01-01

    Bacillus amyloliquefaciens TF28 is a biocontrol endophytic bacterium that is capable of inhibition of a broad range of plant pathogenic fungi. The strain has the potential to be developed into a biocontrol agent for use in agriculture. Here we report the whole-genome shotgun sequence of the strain. The genome size of B. amyloliquefaciens TF28 is 3,987,635 bp which consists of 3754 protein-coding genes, 65 tandem repeat sequences, 47 minisatellite DNA, 2 microsatellite DNA, 63 tRNA, 7rRNA, 6 sRNA, 3 prophage and CRISPR domains.

  4. Bacteriophage biocontrol of foodborne pathogens.

    PubMed

    Kazi, Mustafa; Annapure, Uday S

    2016-03-01

    Bacteriophages are viruses that only infect bacterial cells. Phages are categorized based on the type of their life cycle, the lytic cycle cause lysis of the bacterium with the release of multiple phage particles where as in lysogenic phase the phage DNA is incorporated into the bacterial genome. Lysogeny does not result in lysis of the host. Lytic phages have several potential applications in the food industry as biocontrol agents, biopreservatives and as tools for detecting pathogens. They have also been proposed as alternatives to antibiotics in animal health. Two unique features of phage relevant for food safety are that they are harmless to mammalian cells and high host specificity, keeping the natural microbiota undisturbed. However, the recent approval of bacteriophages as food additives has opened the discussion about 'edible viruses'. This article reviews in detail the application of phages for the control of foodborne pathogens in a process known as "biocontrol".

  5. Antibiosis and bmyB Gene Presence As Prevalent Traits for the Selection of Efficient Bacillus Biocontrol Agents against Crown Gall Disease

    PubMed Central

    Frikha-Gargouri, Olfa; Ben Abdallah, Dorra; Bhar, Ilhem; Tounsi, Slim

    2017-01-01

    This study aimed to improve the screening method for the selection of Bacillus biocontrol agents against crown gall disease. The relationship between the strain biocontrol ability and their in vitro studied traits was investigated to identify the most important factors to be considered for the selection of effective biocontrol agents. In fact, previous selection procedure relying only on in vitro antibacterial activity was shown to be not suitable in some cases. A direct plant-protection strategy was performed to screen the 32 Bacillus biocontrol agent candidates. Moreover, potential in vitro biocontrol traits were investigated including biofilm formation, motility, hemolytic activity, detection of lipopeptide biosynthetic genes (sfp, ituC and bmyB) and production of antibacterial compounds. The obtained results indicated high correlations of the efficiency of the biocontrol with the reduction of gall weight (p = 0.000) and the antibacterial activity in vitro (p = 0.000). Moreover, there was strong correlations of the efficiency of the biocontrol (p = 0.004) and the reduction in gall weight (p = 0.000) with the presence of the bmyB gene. This gene directs the synthesis of the lipopeptide bacillomycin belonging to the iturinic family of lipopeptides. These results were also confirmed by the two-way hierarchical cluster analysis and the correspondence analysis showing the relatedness of these four variables. According to the obtained results a new screening procedure of Bacillus biocontrol agents against crown gall disease could be advanced consisting on two step selection procedure. The first consists on selecting strains with high antibacterial activity in vitro or those harbouring the bmyB gene. Further selection has to be performed on tomato plants in vivo. Moreover, based on the results of the biocontrol assay, five potent strains exhibiting high biocontrol abilities were selected. They were identified as Bacillus subtilis or Bacillus amyloliquefaciens. These

  6. Antibiosis and bmyB Gene Presence As Prevalent Traits for the Selection of Efficient Bacillus Biocontrol Agents against Crown Gall Disease.

    PubMed

    Frikha-Gargouri, Olfa; Ben Abdallah, Dorra; Bhar, Ilhem; Tounsi, Slim

    2017-01-01

    This study aimed to improve the screening method for the selection of Bacillus biocontrol agents against crown gall disease. The relationship between the strain biocontrol ability and their in vitro studied traits was investigated to identify the most important factors to be considered for the selection of effective biocontrol agents. In fact, previous selection procedure relying only on in vitro antibacterial activity was shown to be not suitable in some cases. A direct plant-protection strategy was performed to screen the 32 Bacillus biocontrol agent candidates. Moreover, potential in vitro biocontrol traits were investigated including biofilm formation, motility, hemolytic activity, detection of lipopeptide biosynthetic genes (sfp, ituC and bmyB) and production of antibacterial compounds. The obtained results indicated high correlations of the efficiency of the biocontrol with the reduction of gall weight (p = 0.000) and the antibacterial activity in vitro (p = 0.000). Moreover, there was strong correlations of the efficiency of the biocontrol (p = 0.004) and the reduction in gall weight (p = 0.000) with the presence of the bmyB gene. This gene directs the synthesis of the lipopeptide bacillomycin belonging to the iturinic family of lipopeptides. These results were also confirmed by the two-way hierarchical cluster analysis and the correspondence analysis showing the relatedness of these four variables. According to the obtained results a new screening procedure of Bacillus biocontrol agents against crown gall disease could be advanced consisting on two step selection procedure. The first consists on selecting strains with high antibacterial activity in vitro or those harbouring the bmyB gene. Further selection has to be performed on tomato plants in vivo. Moreover, based on the results of the biocontrol assay, five potent strains exhibiting high biocontrol abilities were selected. They were identified as Bacillus subtilis or Bacillus amyloliquefaciens. These

  7. Biocontrol of the Potato Blackleg and Soft Rot Diseases Caused by Dickeya dianthicola

    PubMed Central

    Raoul des Essarts, Yannick; Cigna, Jérémy; Quêtu-Laurent, Angélique; Caron, Aline; Munier, Euphrasie; Beury-Cirou, Amélie

    2015-01-01

    Development of protection tools targeting Dickeya species is an important issue in the potato production. Here, we present the identification and the characterization of novel biocontrol agents. Successive screenings of 10,000 bacterial isolates led us to retain 58 strains that exhibited growth inhibition properties against several Dickeya sp. and/or Pectobacterium sp. pathogens. Most of them belonged to the Pseudomonas and Bacillus genera. In vitro assays revealed a fitness decrease of the tested Dickeya sp. and Pectobacterium sp. pathogens in the presence of the biocontrol agents. In addition, four independent greenhouse assays performed to evaluate the biocontrol bacteria effect on potato plants artificially contaminated with Dickeya dianthicola revealed that a mix of three biocontrol agents, namely, Pseudomonas putida PA14H7 and Pseudomonas fluorescens PA3G8 and PA4C2, repeatedly decreased the severity of blackleg symptoms as well as the transmission of D. dianthicola to the tuber progeny. This work highlights the use of a combination of biocontrol strains as a potential strategy to limit the soft rot and blackleg diseases caused by D. dianthicola on potato plants and tubers. PMID:26497457

  8. Biocontrol of the Potato Blackleg and Soft Rot Diseases Caused by Dickeya dianthicola.

    PubMed

    Raoul des Essarts, Yannick; Cigna, Jérémy; Quêtu-Laurent, Angélique; Caron, Aline; Munier, Euphrasie; Beury-Cirou, Amélie; Hélias, Valérie; Faure, Denis

    2015-10-23

    Development of protection tools targeting Dickeya species is an important issue in the potato production. Here, we present the identification and the characterization of novel biocontrol agents. Successive screenings of 10,000 bacterial isolates led us to retain 58 strains that exhibited growth inhibition properties against several Dickeya sp. and/or Pectobacterium sp. pathogens. Most of them belonged to the Pseudomonas and Bacillus genera. In vitro assays revealed a fitness decrease of the tested Dickeya sp. and Pectobacterium sp. pathogens in the presence of the biocontrol agents. In addition, four independent greenhouse assays performed to evaluate the biocontrol bacteria effect on potato plants artificially contaminated with Dickeya dianthicola revealed that a mix of three biocontrol agents, namely, Pseudomonas putida PA14H7 and Pseudomonas fluorescens PA3G8 and PA4C2, repeatedly decreased the severity of blackleg symptoms as well as the transmission of D. dianthicola to the tuber progeny. This work highlights the use of a combination of biocontrol strains as a potential strategy to limit the soft rot and blackleg diseases caused by D. dianthicola on potato plants and tubers.

  9. Is the ability of biocontrol fluorescent pseudomonads to produce the antifungal metabolite 2,4-diacetylphloroglucinol really synonymous with higher plant protection?

    PubMed

    Rezzonico, Fabio; Zala, Marcello; Keel, Christoph; Duffy, Brion; Moënne-Loccoz, Yvan; Défago, Geneviève

    2007-01-01

    The antifungal compound 2,4-diacetylphloroglucinol (Phl) contributes to biocontrol in pseudomonads, but whether or not Phl(+) biocontrol pseudomonads display higher plant-protecting activity than Phl(-) biocontrol pseudomonads remains to be demonstrated. This issue was addressed by assessing 230 biocontrol fluorescent pseudomonads selected from a collection of 3132 bacterial isolates obtained from 63 soils worldwide. One-third of the biocontrol pseudomonads were Phl(+) and almost all Phl(+) isolates also produced hydrogen cyanide (HCN). The only Phl(+) HCN(-) strain did harbor hcn genes, but with the deletion of a 134 bp hcnC fragment corresponding to an ADP-binding motif. Statistical analysis of biocontrol isolate distributions indicated that Phl production ability was associated with superior disease suppression activity in the Pythium-cucumber and Fusarium-tomato pathosystems, but this was also the case with HCN production ability. However, HCN significance was not as strong, as indicated both by the comparison of Phl(-) HCN(+) and Phl(-) HCN(-) strains and by correlation analyses. This is the first population-level demonstration of the higher plant-protecting activity of Phl(+) biocontrol pseudomonads in comparison with Phl(-) biocontrol pseudomonads.

  10. Responses of Yeast Biocontrol Agents to Environmental Stress

    PubMed Central

    Sui, Yuan; Wisniewski, Michael; Droby, Samir

    2015-01-01

    Biological control of postharvest diseases, utilizing wild species and strains of antagonistic yeast species, is a research topic that has received considerable attention in the literature over the past 30 years. In principle, it represents a promising alternative to chemical fungicides for the management of postharvest decay of fruits, vegetables, and grains. A yeast-based biocontrol system is composed of a tritrophic interaction between a host (commodity), a pathogen, and a yeast species, all of which are affected by environmental factors such as temperature, pH, and UV light as well as osmotic and oxidative stresses. Additionally, during the production process, biocontrol agents encounter various severe abiotic stresses that also impact their viability. Therefore, understanding the ecological fitness of the potential yeast biocontrol agents and developing strategies to enhance their stress tolerance are essential to their efficacy and commercial application. The current review provides an overview of the responses of antagonistic yeast species to various environmental stresses, the methods that can be used to improve stress tolerance and efficacy, and the related mechanisms associated with improved stress tolerance. PMID:25710368

  11. Postharvest Biocontrol: Introspection and Paradigm Shifts

    USDA-ARS?s Scientific Manuscript database

    The use of postharvest biocontrol agents as an alternative to the synthetic, chemical fungicides on a widespread basis has many constraints. During the last twenty years, the field of postharvest biocontrol research has significantly grown and developed and seen the creation of several products. Des...

  12. Enhancement of Biocontrol Activities and Cyclic Lipopeptides Production by Chemical Mutagenesis of Bacillus subtilis XF-1, a Biocontrol Agent of Plasmodiophora brassicae and Fusarium solani.

    PubMed

    Li, Xing-Yu; Yang, Jing-Jing; Mao, Zi-Chao; Ho, Hon-Hing; Wu, Yi-Xing; He, Yue-Qiu

    2014-12-01

    Bacillus subtilis XF-1 has been used as a biocontrol agent of clubroot disease of crucifers infected by Plasmodiophora brassicae, an obligate pathogen. In order to maximize the growth inhibition of the pathogen, random mutagenesis using N-methyl-N'-nitro-N-nitrosoguanidine was applied to strain XF-1. The efficacy of 226 selected mutants was assessed against the growth of an indicator fungal pathogen: Fusarium solani using agar plate assay and the disruptive effects on the resting spores of P. brassicae. Four mutants exhibited inhibition activity significantly higher than the wild type. The cell extracts of these mutants and the XF-1 were subjected to matrix-assisted laser desorption ionization-time of flight mass spectra analysis, and three families of cyclic lipopeptides (CLPs) fengycin, surfactin and iturin were identified from the parental strain and the screened mutants. However, the relative contents and compound diversity changed after mutagenesis, and there was slight variation in the surfactin and fengycin. Notably, only 5 iturin components were discovered from the wild strain XF-1, but 13 were obtained from the mutant strains, and the relative CLPs contents of all mutant strains increased substantially. The results suggested that CLPs might be one of main biocontrol mechanisms of the clubroot disease by XF-1. The 4 mutants are far more effective than the parental strain, and they would be promising biocontrol candidates not only against P. brassicae but probably other plant diseases caused by fungi.

  13. [Correction of autonomic reactions parameters in organism of cosmonaut with adaptive biocontrol method

    NASA Technical Reports Server (NTRS)

    Kornilova, L. N.; Cowings, P. S.; Toscano, W. B.; Arlashchenko, N. I.; Korneev, D. Iu; Ponomarenko, A. V.; Salagovich, S. V.; Sarantseva, A. V.; Kozlovskaia, I. B.

    2000-01-01

    Presented are results of testing the method of adaptive biocontrol during preflight training of cosmonauts. Within the MIR-25 crew, a high level of controllability of the autonomous reactions was characteristic of Flight Commanders MIR-23 and MIR-25 and flight Engineer MIR-23, while Flight Engineer MIR-25 displayed a weak intricate dependence of these reactions on the depth of relaxation or strain.

  14. [Correction of autonomic reactions parameters in organism of cosmonaut with adaptive biocontrol method].

    PubMed

    Kornilova, L N; Cowings, P S; Toscano, W B; Arlashchenko, N I; Korneev, D Iu; Ponomarenko, A V; Salagovich, S V; Sarantseva, A V; Kozlovskaia, I B

    2000-01-01

    Presented are results of testing the method of adaptive biocontrol during preflight training of cosmonauts. Within the MIR-25 crew, a high level of controllability of the autonomous reactions was characteristic of Flight Commanders MIR-23 and MIR-25 and flight Engineer MIR-23, while Flight Engineer MIR-25 displayed a weak intricate dependence of these reactions on the depth of relaxation or strain.

  15. Pantoea applied genomics to understand and improve biocontrol activity against fire blight

    USDA-ARS?s Scientific Manuscript database

    Pantoea agglomerans and P. vagans (ex. Erwinia herbicola) are common epiphytes of pome fruit flowers and three strains (E325, P10c, C9-1) have been commercially developed as effective biocontrol products for managing fire blight (Erwinia amylovora). Antibiotics as a standard, reliable chemical optio...

  16. [Correction of autonomic reactions parameters in organism of cosmonaut with adaptive biocontrol method

    NASA Technical Reports Server (NTRS)

    Kornilova, L. N.; Cowings, P. S.; Toscano, W. B.; Arlashchenko, N. I.; Korneev, D. Iu; Ponomarenko, A. V.; Salagovich, S. V.; Sarantseva, A. V.; Kozlovskaia, I. B.

    2000-01-01

    Presented are results of testing the method of adaptive biocontrol during preflight training of cosmonauts. Within the MIR-25 crew, a high level of controllability of the autonomous reactions was characteristic of Flight Commanders MIR-23 and MIR-25 and flight Engineer MIR-23, while Flight Engineer MIR-25 displayed a weak intricate dependence of these reactions on the depth of relaxation or strain.

  17. Metschnikowia fructicola, a new ascosporic yeast with potential for biocontrol of postharvest fruit rots.

    PubMed

    Kurtzman, C P; Droby, S

    2001-11-01

    A new ascosporic yeast, Metschnikowia fructicola (type strain NRRL Y-27328, CBS 8853), is described and was isolated from grapes grown in central Israel. Preliminary tests indicate the new species has biocontrol activity against Botrytis rot of stored grapes. Phylogenetic analysis of domain D1/D2 26S rDNA sequences showed M. fructicola to be a sister species of M. pulcherrima.

  18. Heat shock improves stress tolerance and biocontrol performance of Rhodotorula mucilaginosa

    USDA-ARS?s Scientific Manuscript database

    Biological control of postharvest diseases, utilizing wild species and strains of antagonistic yeasts, is a research topic and eco-friendly management approach that has received considerable attention. A few yeast-based biocontrol products are available in the market. Ecological fitness, stress to...

  19. Biocontrol of verticillium wilt and colonization of cotton plants by an endophytic bacterial isolate.

    PubMed

    Li, C-H; Shi, L; Han, Q; Hu, H-L; Zhao, M-W; Tang, C-M; Li, S-P

    2012-09-01

    To explore biocontrol potential of 39 DAEB isolates (doubly antagonistic towards both Verticillium dahliae Kleb and Fusarium oxysporum) against verticillium wilt of cotton and to elucidate colonization and category characteristics of an endophytic bacterium with significant biocontrol activity. Thirty-nine antagonistic endophytic bacteria strains were tested for their ability to control verticillium wilt in cotton plants caused by a defoliating pathotype of V. dahliae 107 in cotton under controlled conditions. The biocontrol trial revealed that an endophytic bacterium, designated HA02, showed a significant biocontrol activity to V. dahliae 107. After cotton seedlings were inoculated with a gfp gene-tagged HA02 (HA02-gfp), HA02-gfp populations were higher in the root than in the stem; in addition, the HA02-gfp was distributed in the maturation zone of cotton root. Furthermore, HA02-gfp also colonized seedlings of maize, rape and soybean after the bacteria inoculation. Phylogenetic trees based on 16S rDNA sequences combined with morphological, physiological and identification showed that the bacterium belongs to the Enterobacter genus. Our results showed that only 1 of 39 DAEB isolates demonstrated more efficient biocontrol potential towards V. dahliae 107 in greenhouse and field trials. HA02-gfp mainly colonized cotton in roots. In addition, we quantitatively observed HA02 colonization in other hosts. HA02 belongs to the Enterobacter genus. This is the first study on biocontrol to defoliating pathotype of V. dahliae Kleb by endophytic bacteria. The HA02 showed effective biocontrol to V. dahliae 107 in greenhouse and field trials. Furthermore, we assessed the quantitative and qualitative colonization of HA02 in cotton seedlings. Our study provides basic information to further explore managing strategies to control this critical disease. © 2012 The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  20. Impact of a Recombinant Biocontrol Bacterium, Pseudomonas fluorescens pc78, on Microbial Community in Tomato Rhizosphere.

    PubMed

    Kong, Hyun Gi; Kim, Nam Hee; Lee, Seung Yeup; Lee, Seon-Woo

    2016-04-01

    Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer.

  1. Impact of a Recombinant Biocontrol Bacterium, Pseudomonas fluorescens pc78, on Microbial Community in Tomato Rhizosphere

    PubMed Central

    Kong, Hyun Gi; Kim, Nam Hee; Lee, Seung Yeup; Lee, Seon-Woo

    2016-01-01

    Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer. PMID:27147933

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

    PubMed

    Li, Boqiang; Peng, Huaimin; Tian, Shiping

    2016-01-01

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

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

    PubMed Central

    Li, Boqiang; Peng, Huaimin; Tian, Shiping

    2016-01-01

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

  4. Novel Routes for Improving Biocontrol Activity of Bacillus Based Bioinoculants

    PubMed Central

    Wu, Liming; Wu, Hui-Jun; Qiao, Junqing; Gao, Xuewen; Borriss, Rainer

    2015-01-01

    Biocontrol (BC) formulations prepared from plant-growth-promoting bacteria are increasingly applied in sustainable agriculture. Especially inoculants prepared from endospore-forming Bacillus strains have been proven as efficient and environmental-friendly alternative to chemical pesticides due to their long shelf life, which is comparable with that of agrochemicals. However, these formulations of the first generation are sometimes hampered in their action and do not fulfill in each case the expectations of the appliers. In this review we use the well-known plant-associated Bacillus amyloliquefaciens type strain FZB42 as example for the successful application of different techniques offered today by comparative, evolutionary and functional genomics, site-directed mutagenesis and strain construction including marker removal, for paving the way for preparing a novel generation of BC agents. PMID:26696998

  5. Assessments of the microbial biocontrol potential of novel strains of Beauveria bassiana s.l. against greenhouse shore fly, Scatella tenuicosta: virulence, mass production capacity, and effects on shore fly reproduction

    USDA-ARS?s Scientific Manuscript database

    The biological control potential of three strains of Beauveria bassiana s.l. originally isolated from the shore fly Scatella ternuicosta was assessed in a series of laboratory bioassays. Comparisons were made to two commercially available strains of B. bassiana. Two of the new strains proved 27–67 t...

  6. Effect of heat shock treatment on stress tolerance and biocontrol efficacy of biocontrol yeasts

    USDA-ARS?s Scientific Manuscript database

    Several different species of yeasts have been used as biocontrol agents against postharvest diseases of fruits and vegetables. Our current research is directed to develop a better understanding of yeast biology in relation to biocontrol activity and to develop strategies to improve the efficacy of ...

  7. Brazilian isolates of Clonostachys rosea: colonization under different temperature and moisture conditions and temporal dynamics on strawberry leaves.

    PubMed

    Cota, L V; Maffia, L A; Mizubuti, E S G

    2008-03-01

    In a research programme for managing diseases caused by Botryis cinerea, four isolates of the antagonistic fungus Clonostachys rosea (Cr) were obtained from different ecosystems in Brazil. We studied ecological requirements for the colonization of strawberry leaves by these isolates. Temperature effects on both mycelial growth in vitro and leaf colonization by Cr were studied. At 10 degrees C, growth on potato dextrose agar and colonization of leaf discs were poor. Optimum temperature for mycelial growth and leaf colonization was around 25 degrees C. The isolates were applied to leaves which were exposed to 0-48 h intervals of moisture. They were also applied to leaves which remained from 0 to 36 h without wetness. All isolates efficiently colonized leaves, regardless of moisture interval or the delay to begin wetness. Although all isolates survived in green leaves of whole plants, colonization decreased throughout a 49-day period. Brazilian isolates of Cr can establish and colonize strawberry leaves under a wide range of temperature and moisture conditions. It is expected that the Brazilian isolates of Cr will establish efficiently in strawberry leaves where they can compete with B. cinerea.

  8. Effect of Trichoderma harzianum on maize rhizosphere microbiome and biocontrol of Fusarium Stalk rot.

    PubMed

    Saravanakumar, Kandasamy; Li, Yaqian; Yu, Chuanjin; Wang, Qiang-Qiang; Wang, Meng; Sun, Jianan; Gao, Jin-Xin; Chen, Jie

    2017-05-11

    Fusarium stalk rot (FSR) caused by Fusarium graminearum (FG) significantly affects the productivity of maize grain crops. Application of agrochemicals to control the disease is harmful to environment. In this regard, use of biocontrol agent (BCA) is an alternative to agrochemicals. Although Trichoderma species are known as BCA, the selection of host-pathogen specific Trichoderma is essential for the successful field application. Hence, we screened a total of 100 Trichoderma isolates against FG, selected Trichoderma harzianum (CCTCC-RW0024) for greenhouse experiments and studied its effect on changes of maize rhizosphere microbiome and biocontrol of FSR. The strain CCTCC-RW0024 displayed high antagonistic activity (96.30%), disease reduction (86.66%), biocontrol-related enzyme and gene expression. The root colonization of the strain was confirmed by eGFP tagging and qRT-PCR analysis. Pyrosequencing revealed that exogenous inoculation of the strain in maize rhizosphere increased the plant growth promoting acidobacteria (18.4%), decreased 66% of FG, and also increased the plant growth. In addition, metabolites of this strain could interact with pathogenicity related transcriptional cofactor FgSWi6, thereby contributing to its inhibition. It is concluded that T. harzianum strain CCTCC-RW0024 is a potential BCA against FSR.

  9. Draft Genome Sequences of Pseudomonas fluorescens Strains PA4C2 and PA3G8 and Pseudomonas putida PA14H7, Three Biocontrol Bacteria against Dickeya Phytopathogens

    PubMed Central

    Cigna, Jérémy; Raoul des Essarts, Yannick; Mondy, Samuel; Hélias, Valérie; Beury-Cirou, Amélie

    2015-01-01

    Pseudomonas fluorescens strains PA4C2 and PA3G8 and Pseudomonas putida strain PA14H7 were isolated from potato rhizosphere and show an ability to inhibit the growth of Dickeya phytopathogens. Here, we report their draft genome sequences, which provide a basis for understanding the molecular mechanisms involved in antibiosis against Dickeya. PMID:25635023

  10. Preliminary in vitro insights into the use of natural fungal pathogens of leaf-cutting ants as biocontrol agents.

    PubMed

    Folgarait, Patricia; Gorosito, Norma; Poulsen, Michael; Currie, Cameron R

    2011-09-01

    Leaf-cutting ants are one of the main herbivores of the Neotropics, where they represent an important agricultural pest. These ants are particularly difficult to control because of the complex network of microbial symbionts. Leaf-cutting ants have traditionally been controlled through pesticide application, but there is a need for alternative, more environmentally friendly, control methods such as biological control. Potential promising biocontrol candidates include the microfungi Escovopsis spp. (anamorphic Hypocreales), which are specialized pathogens of the fungi the ants cultivate for food. These pathogens are suppressed through ant behaviors and ant-associated antibiotic-producing Actinobacteria. In order to be an effective biocontrol agent, Escovopsis has to overcome these defenses. Here, we evaluate, using microbial in vitro assays, whether defenses in the ant-cultivated fungus strain (Leucoagaricus sp.) and Actinobacteria from the ant pest Acromyrmex lundii have the potential to limit the use of Escovopsis in biocontrol. We also explore, for the first time, possible synergistic biocontrol between Escovopsis and the entomopathogenic fungus Lecanicillium lecanii. All strains of Escovopsis proved to overgrow A. lundii cultivar in less than 7 days, with the Escovopsis strain isolated from a different leaf-cutting ant species being the most efficient. Escovopsis challenged with a Streptomyces strain isolated from A. lundii did not exhibit significant growth inhibition. Both results are encouraging for the use of Escovopsis as a biocontrol agent. Although we found that L. lecanii can suppress the growth of the cultivar, it also had a negative impact on Escovopsis, making the success of simultaneous use of these two fungi for biocontrol of A. lundii questionable.

  11. Non-aflatoxigenic Aspergillus flavus as potential biocontrol agents to reduce aflatoxin contamination in peanuts harvested in Northern Argentina.

    PubMed

    Alaniz Zanon, María Silvina; Barros, Germán Gustavo; Chulze, Sofía Noemí

    2016-08-16

    Biological control is one of the most promising strategies for preventing aflatoxin contamination in peanuts at field stage. A population of 46 native Aspergillus flavus nonaflatoxin producers were analysed based on phenotypic, physiological and genetic characteristics. Thirty-three isolates were characterized as L strain morphotype, 3 isolates as S strain morphotype, and 10 isolates did not produce sclerotia. Only 11 of 46 non-aflatoxigenic isolates did not produce cyclopiazonic acid. The vegetative compatibility group (VCG) diversity index for the population was 0.37. For field trials we selected the non-aflatoxigenic A. flavus AR27, AR100G and AFCHG2 strains. The efficacy of single and mixed inocula as potential biocontrol agents in Northern Argentina was evaluated through a 2-year study (2014-2015). During the 2014 peanut growing season, most of the treatments reduced the incidence of aflatoxigenic strains in both soil and peanut kernel samples, and no aflatoxin was detected in kernels. During the 2015 growing season, there was a reduction of aflatoxigenic strains in kernel samples from the plots treated with the potential biocontrol agents. Reductions of aflatoxin contamination between 78.36% and 89.55% were observed in treated plots in comparison with the un-inoculated control plots. This study provides the first data on aflatoxin biocontrol based on competitive exclusion in the peanut growing region of Northern Argentina, and proposes bioproducts with potential use as biocontrol agents. Copyright © 2016. Published by Elsevier B.V.

  12. From biocontrol to cancer, probiotics and beyond.

    PubMed

    Dunne, Colum

    2013-01-01

    This invited commentary covers the period 1997-2012 and has seen changes in terminology that progressed from "basic" and "applied" to "translational" research. In the context of Bioengineered, these changes map readily onto the processes of identifying microbial characteristics appropriate for specific applications, isolation of suitable cultures, strain or genome manipulation and exploitation of these or their metabolomes across a range of settings.   To a great degree, this commentary and my career reflect an engagement with molecular microbiology and the trialling of bacteria and derived constructs in applications ranging from intensive-scale crop protection to amelioration of gastrointestinal disease. This engagement began with laboratory and field evaluations of biocontrol, specifically use of pseudomonads effective against nematode and fungal plant pathogens, characterization of mechanisms mediating beneficial effects of probiotic lactobacilli and bifidobacteria and assessment of functional foods in multinational clinical trials relating to inflammatory bowel disease. Subsequent work focused on (1) intellectual property (IP)-based medical devices for localized delivery of systemically toxic and gene cancer therapies; (2) growth of the science base supporting expansion of a multinational business including company acquisitions; (3) complementing existing inter-institutional research capabilities through development of a national industry-led collaboration; and, most recently, (4) strategic research programs at Ireland's newest medical school. My activities as outlined above parallel two distinct aspects of translational research: (1) involvement in knowledge-driven (commercial and research) organizations that brought together necessary resources and infrastructure and (2) availability of scale research funding from European Framework and Irish national programs.

  13. Indigenous bacteria may interfere with the biocontrol of plant diseases

    NASA Astrophysics Data System (ADS)

    Someya, Nobutaka; Akutsu, Katsumi

    2009-06-01

    Prodigiosin is a reddish antibiotic pigment that plays an important role in the biocontrol of plant diseases by the bacterium Serratia marcescens. However, its activity is unstable under agricultural conditions; further, it can be degraded by various environmental factors. To examine the effect of epiphytic microbes on the stability of prodigiosin used for biological control processes, we collected a total of 1,280 bacterial isolates from the phylloplane of cyclamen and tomato plants. Approximately 72% of the bacterial strains isolated from the cyclamen plants and 66% of those isolated from the tomato plants grew on minimal agar medium containing 100 μg ml-1 prodigiosin. Certain isolates obtained from both plant species exhibited prodigiosin-degrading activity. We compared the 16S rRNA gene sequences derived from the isolates with sequences in a database. The comparison revealed that the sequences determined for the prodigiosin-degrading isolates were homologous to those of the genera Pseudomonas, Caulobacter, Rhizobium, Sphingomonas, Janthinobacterium, Novosphingobium, and Rathayibacter. These results indicate that indigenous epiphytic microorganisms may interfere with the interaction between plant pathogens and biocontrol agents by degrading the antibiotics produced by the agents.

  14. Identification of Biocontrol Agents to Control the Fungal Pathogen, Geomyces destructans, in Bats

    NASA Astrophysics Data System (ADS)

    Braunstein, S.; Cheng, T.

    2013-12-01

    The fungal pathogen Geomyces destructans (Gd) causes the disease White-nose Syndrome (WNS) in bats and is estimated to have killed millions of bats since its emergence in North America in 2006. Gd is predicted to cause the local extinction of at least three bat species if rates of decline continue unabated. Given the devastating impacts of Gd to bat populations, identifying a viable method for controlling the pathogen is pertinent for conservation of affected bat species. Our work focuses on identifying naturally-occurring skin bacteria on bats that are antagonistic to Gd that could potentially be used as a biocontrol. We cultured bacteria from skin swabs taken from wild bats (Myotis lucifugus, Eptesicus fuscus, Myotis sodalis, Perimyotis subflavus). We conducted challenge experiments to identify bacterial strains that inhibited Gd growth. Bacteria that exhibited antifungal properties were identified using 16S and gyrB markers. Our methods identified several bacteria in the Pseudomonas fluorescens complex as potential biocontrol agents. Future work will continue to test the viability of these bacteria as biocontrol agents via experimental treatments with live captive bats. The failure of previous non-biocontrol methods highlights the importance of developing these bacteria as a biologically-friendly method for controlling Gd. A bat infected with Geomyces destructans. Photo by West Virginia Division of Natural Resources Bacterial culture from the swab of a bat's wings

  15. Potential of osmoadaptation for improving Pantoea agglomerans E325 as biocontrol agent for fire blight of apple and pear

    USDA-ARS?s Scientific Manuscript database

    Pantoea agglomerans biocontrol strain E325 is the active ingredient in a commercial product for fire blight, a destructive disease of apple and pear initiated by Erwinia amylovora in flowers. Osmoadaptation, involving the combination of saline osmotic stress and osmolyte amendment to growth media, w...

  16. Impacts of biocontrol products on Rhizoctonia disease of potato and soil microbial communities, and their persistence in soil

    USDA-ARS?s Scientific Manuscript database

    Four commercial biocontrol formulations (Bacillus subtilis GB03, Burkholderia ambifaria type Wisconsin isolate J82, Trichoderma virens Gl-21, and Trichoderma harzianum strain T-22), a chemical seed treatment (Topsin, mancozeb, and cymoxanil mixture, TMC), and a combination chemical/biological treatm...

  17. Biocontrol agents promote growth of potato pathogens, depending on environmental conditions.

    PubMed

    Cray, Jonathan A; Connor, Mairéad C; Stevenson, Andrew; Houghton, Jonathan D R; Rangel, Drauzio E N; Cooke, Louise R; Hallsworth, John E

    2016-05-01

    There is a pressing need to understand and optimize biological control so as to avoid over-reliance on the synthetic chemical pesticides that can damage environmental and human health. This study focused on interactions between a novel biocontrol-strain, Bacillus sp. JC12GB43, and potato-pathogenic Phytophthora and Fusarium species. In assays carried out in vitro and on the potato tuber, the bacterium was capable of near-complete inhibition of pathogens. This Bacillus was sufficiently xerotolerant (water activity limit for growth = 0.928) to out-perform Phytophthora infestans (~0.960) and challenge Fusarium coeruleum (~0.847) and Fusarium sambucinum (~0.860) towards the lower limits of their growth windows. Under some conditions, however, strain JC12GB43 stimulated proliferation of the pathogens: for instance, Fusarium coeruleum growth-rate was increased under chaotropic conditions in vitro (132 mM urea) by >100% and on tubers (2-M glycerol) by up to 570%. Culture-based assays involving macromolecule-stabilizing (kosmotropic) compatible solutes provided proof-of-principle that the Bacillus may provide kosmotropic metabolites to the plant pathogen under conditions that destabilize macromolecular systems of the fungal cell. Whilst unprecedented, this finding is consistent with earlier reports that fungi can utilize metabolites derived from bacterial cells. Unless the antimicrobial activities of candidate biocontrol strains are assayed over a full range of field-relevant parameters, biocontrol agents may promote plant pathogen infections and thereby reduce crop yields. These findings indicate that biocontrol activity, therefore, ought to be regarded as a mode-of-behaviour (dependent on prevailing conditions) rather than an inherent property of a bacterial strain. © 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  18. A new method to evaluate the biocontrol potential of single spore isolates of fungal entomopathogens

    PubMed Central

    Posada, Francisco J.; Vega, Fernando E.

    2005-01-01

    Fifty Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) strains isolated from the coffee berry borer were used to develop a novel screening method aimed at selecting strains with the highest biocontrol potential. The screening method is based on percent insect mortality, average survival time, mortality distribution, percent spore germination, fungal life cycle duration, and spore production on the insect. Based on these parameters, only 11 strains merited further study. The use of a sound scientific protocol for the selection of promising fungal entomopathogens should lead to more efficient use of time, labor, and financial resources in biological control programs. PMID:17119619

  19. A new method to evaluate the biocontrol potential of single spore isolates of fungal entomopathogens.

    PubMed

    Posada, Francisco J; Vega, Fernando E

    2005-12-06

    Fifty Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) strains isolated from the coffee berry borer were used to develop a novel screening method aimed at selecting strains with the highest biocontrol potential. The screening method is based on percent insect mortality, average survival time, mortality distribution, percent spore germination, fungal life cycle duration, and spore production on the insect. Based on these parameters, only 11 strains merited further study. The use of a sound scientific protocol for the selection of promising fungal entomopathogens should lead to more efficient use of time, labor, and financial resources in biological control programs.

  20. Evaluation of economically feasible, natural plant extract-based microbiological media for producing biomass of the dry rot biocontrol strain Pseudomonas fluorescens P22Y05 in liquid culture

    USDA-ARS?s Scientific Manuscript database

    The production of microbial biomass in liquid media often represents an indispensable step in the research and development of bacterial and fungal strains. Costs of commercially prepared nutrient media or purified media components, however, can represent a significant hurdle to conducting research i...

  1. Hrp mutant bacteria as biocontrol agents: toward a sustainable approach in the fight against plant pathogenic bacteria.

    PubMed

    Hanemian, Mathieu; Zhou, Binbin; Deslandes, Laurent; Marco, Yves; Trémousaygue, Dominique

    2013-10-01

    Sustainable agriculture necessitates development of environmentally safe methods to protect plants against pathogens. Among these methods, application of biocontrol agents has been efficiently used to minimize disease development. Here we review current understanding of mechanisms involved in biocontrol of the main Gram-phytopathogenic bacteria-induced diseases by plant inoculation with strains mutated in hrp (hypersensitive response and pathogenicity) genes. These mutants are able to penetrate plant tissues and to stimulate basal resistance of plants. Novel protection mechanisms involving the phytohormone abscisic acid appear to play key roles in the biocontrol of wilt disease induced by Ralstonia solanacearum in Arabidopsis thaliana. Fully understanding these mechanisms and extending the studies to other pathosystems are still required to evaluate their importance in disease protection.

  2. Biocontrol activity of a cold-adapted yeast from Tibet against gray mold in cherry tomato and its action mechanism.

    PubMed

    Hu, Hao; Wisniewski, Michael E; Abdelfattah, Ahmed; Zheng, Xiaodong

    2017-07-01

    Cold-adapted biocontrol yeast was selected from four yeast isolates from Tibet against gray mold of cherry tomato in cold storage. The strain numbered LB2 showed the best biocontrol activity and identified as Cryptococcus laurentii. Competition for nutrient, space, and induced fruit resistance was also its antagonistic mechanism. Compared with C. laurentii from sea-level place, the reason why LB2 had a better biocontrol activity was studied. More trehalose and proline in cell of LB2 made it exhibit a better cellular activity at low temperature, such as higher population dynamics in the wounds of cherry tomato and more biocontrol-related enzyme secretion, chitinase and β-glucanase. The better oxidative stress tolerance was another characteristic of LB2. Maybe because of the ideal culture condition, there was no obvious difference between these two yeasts in the growth in vitro test at low temperature. Although the same phenomenon existed in the low pH stress test, LB2 still had higher cell concentration under this stress. Comparative transcriptomics method was also applied to analyze the cell activity of LB2 and C. laurentii at different temperatures. The results showed that more active response in the intracellular structure and intracellular metabolic process to cold temperature made LB2 had a better activity. The present study indicated a possibility to select cold-adapted biocontrol yeast from Tibet and also showed its primary action mechanism.

  3. Commercialization of postharvest biocontrol: barriers and opportunities

    USDA-ARS?s Scientific Manuscript database

    The past twenty years has seen the field of postharvest biocontrol evolve from an unknown entity with one or two novel reports in the literature to a sophisticated science with strong research programs worldwide, hundreds of publications, patented technologies, and now several commercial products. ...

  4. Experiments with Biocontrol of Alternaria alternata

    USDA-ARS?s Scientific Manuscript database

    A biocontrol yeast (WRL-76) developed by USDA-ARS at the Western Regional Research lab was evaluated for control of Alternaria alternata on pistachio. The experiment was conducted over a four year period in a Madera Co., CA orchard. Counts of damaged vs. intact fruit clusters were taken on 100 contr...

  5. Biocontrol of Burkholderia cepacia complex bacteria and bacterial phytopathogens by Bdellovibrio bacteriovorus.

    PubMed

    McNeely, Damian; Chanyi, Ryan M; Dooley, James S; Moore, John E; Koval, Susan F

    2017-04-01

    Bdellovibrio and like organisms are predatory bacteria that have the unusual property of using the cytoplasmic constituents of other Gram-negative bacteria as nutrients. These predators may thus provide an alternative approach to the biocontrol of human and plant pathogens. Predators were isolated on Burkholderia cenocepacia K56-2 and J2315 as prey cells, in enrichment cultures with soil and sewage. Three isolates (DM7C, DM8A, and DM11A) were identified as Bdellovibrio bacteriovorus on the basis of morphology, a periplasmic life cycle, and 16S rRNA gene sequencing. The prey range of these isolates was tested on Burkholderia cepacia complex bacteria and several phytopathogenic bacteria of agricultural importance. Of 31 strains of the Burkholderia cepacia complex tested, only 4 were resistant to predation by strain DM7C. A subset of 9 of the prey tested were also susceptible to strains DM8A and DM11A. Of 12 phytopathogens tested, 4 were resistant to strains DM7C and DM8A, and only 2 were resistant to strain DM11A. Thus, Bdellovibrio bacteriovorus strains retrieved from environmental samples on 2 Burkholderia cenocepacia isolates from cystic fibrosis patients did not distinguish in their prey range between other isolates of that pathogen or phytopathogens. Such strains hold promise as potential wide-spectrum biocontrol agents.

  6. Genetic diversity and biocontrol potential of fluorescent pseudomonads producing phloroglucinols and hydrogen cyanide from Swiss soils naturally suppressive or conducive to Thielaviopsis basicola-mediated black root rot of tobacco.

    PubMed

    Ramette, Alban; Moënne-Loccoz, Yvan; Défago, Geneviève

    2006-03-01

    Pseudomonas populations producing the biocontrol compounds 2,4-diacetylphloroglucinol (Phl) and hydrogen cyanide (HCN) were found in the rhizosphere of tobacco both in Swiss soils suppressive to Thielaviopsis basicola and in their conducive counterparts. In this study, a collection of Phl+ HCN+Pseudomonas isolates from two suppressive and two conducive soils were used to assess whether suppressiveness could be linked to soil-specific properties of individual pseudomonads. The isolates were compared based on restriction analysis of the biocontrol genes phlD and hcnBC, enterobacterial repetitive intergenic consensus (ERIC)-PCR profiling and their biocontrol ability. Restriction analyses of phlD and hcnBC yielded very concordant relationships between the strains, and suggested significant population differentiation occurring at the soil level, regardless of soil suppressiveness status. This was corroborated by high strain diversity (ERIC-PCR) within each of the four soils and among isolates harboring the same phlD or hcnBC alleles. No correlation was found between the origin of the isolates and their biocontrol activity in vitro and in planta. Significant differences in T. basicola inhibition were however evidenced between the isolates when they were grouped according to their biocontrol alleles. Moreover, two main Pseudomonas lineages differing by the capacity to produce pyoluteorin were evidenced in the collection. Thus, Phl+ HCN+ pseudomonads from suppressive soils were not markedly different from those from nearby conducive soils. Therefore, as far as biocontrol pseudomonads are concerned, this work yields the hypothesis that the suppressiveness of Swiss soils may rely on the differential effects of environmental factors on the expression of key biocontrol genes in pseudomonads rather than differences in population structure of biocontrol Pseudomonas subcommunities or the biocontrol potential of individual Phl+ HCN+ pseudomonad strains.

  7. Effect of culture media and pH on the biomass production and biocontrol efficacy of a Metschnikowia pulcherrima strain to be used as a biofungicide for postharvest disease control.

    PubMed

    Spadaro, D; Ciavorella, A; Dianpeng, Z; Garibaldi, A; Gullino, M L

    2010-02-01

    Few strains of Metschnikowia pulcherrima (Pitt) M.W. Miller are under development for control of postharvest pathogens on fruit. A substrate was developed to optimize the biomass production of M. pulcherrima strain BIO126. Different complex nutrient sources, with or without pH control, were tested. Growth in yeast extract provided at concentrations > or =30 g*L-1 yielded the highest biomass. The addition of two carbon sources, D-mannitol and L-sorbose, at 5 g*L-1 each, significantly improved yeast growth. The greatest amount of yeast growth occurred when pH values of the medium ranged from 5.0 to 7.5. A combination of yeast extract, D-mannitol, and L-sorbose (YEMS), probably with diauxic utilization, showed a synergistic effect, widening the exponential phase (maximum specific growth rate of 0.45 h-1) and increasing the final cell number (1.5 x 109 cells*mL-1) and dry biomass (6.0 g*L-1) in well-controlled batch fermentation. In efficacy trials on 'Golden Delicious' apples, M. pulcherrima grown in YEMS effectively reduced incidence and severity of Botrytis cinerea (51.1% and 70.8%, respectively) and Penicillium expansum (41.7% and 14.0%, respectively). Also on 'Gala' apples, the best reduction of grey and blue mould incidence was obtained with cells grown in YEMS (58.1% and 50.5%, respectively).

  8. Evaluation of the Biocontrol Potential of Purpureocillium lilacinum QLP12 against Verticillium dahliae in Eggplant.

    PubMed

    Lan, Xingjie; Zhang, Jing; Zong, Zhaofeng; Ma, Qing; Wang, Yang

    2017-01-01

    A fungus with broad spectrum antifungal activity was isolated from the soil in Qinling Mountain, Shaanxi Province, in China. The fungus was identified as Purpureocillium lilacinum based on ITS rDNA gene analysis. The strain, coded as QLP12, showed high inhibition activity on fungal mycelium growth in vitro, especially to Mucor piriformis, Trichothecium roseum, Rhizoctonia solani, and Verticillium dahliae, and its potential for biocontrol efficacy of eggplant. Verticillium wilt disease caused by Verticillium dahliae among 10 fungal species tested was explored. In greenhouse experiments, QLP12 showed an excellent growth-promoting effect on eggplant seed germination (76.7%), bud growth (79.4%), chlorophyll content (47.83%), root activity (182.02%), and so on. QLP12 can colonize the eggplant interior and also develop in rhizosphere soil. In greenhouse, the incidence of Verticillium wilt decreased by 83.82% with pretreated QLP12 fermentation broth in the soil. In the field, QLP12 showed prominent biocontrol effects on Verticillium wilt by reducing the disease index over the whole growth period, a decline of 40.1%. This study showed that the strain QLP12 is not only an effective biocontrol agent for controlling Verticillium wilt of eggplant, but also a plant growth-promoting fungus that deserves to be further developed.

  9. Evaluation of the Biocontrol Potential of Purpureocillium lilacinum QLP12 against Verticillium dahliae in Eggplant

    PubMed Central

    Zhang, Jing; Zong, Zhaofeng

    2017-01-01

    A fungus with broad spectrum antifungal activity was isolated from the soil in Qinling Mountain, Shaanxi Province, in China. The fungus was identified as Purpureocillium lilacinum based on ITS rDNA gene analysis. The strain, coded as QLP12, showed high inhibition activity on fungal mycelium growth in vitro, especially to Mucor piriformis, Trichothecium roseum, Rhizoctonia solani, and Verticillium dahliae, and its potential for biocontrol efficacy of eggplant. Verticillium wilt disease caused by Verticillium dahliae among 10 fungal species tested was explored. In greenhouse experiments, QLP12 showed an excellent growth-promoting effect on eggplant seed germination (76.7%), bud growth (79.4%), chlorophyll content (47.83%), root activity (182.02%), and so on. QLP12 can colonize the eggplant interior and also develop in rhizosphere soil. In greenhouse, the incidence of Verticillium wilt decreased by 83.82% with pretreated QLP12 fermentation broth in the soil. In the field, QLP12 showed prominent biocontrol effects on Verticillium wilt by reducing the disease index over the whole growth period, a decline of 40.1%. This study showed that the strain QLP12 is not only an effective biocontrol agent for controlling Verticillium wilt of eggplant, but also a plant growth-promoting fungus that deserves to be further developed. PMID:28303252

  10. Isolation of Secondary Metabolites from the Soil-Derived Fungus Clonostachys rosea YRS-06, a Biological Control Agent, and Evaluation of Antibacterial Activity.

    PubMed

    Zhai, Ming-Ming; Qi, Feng-Ming; Li, Jie; Jiang, Chun-Xiao; Hou, Yue; Shi, Yan-Ping; Di, Duo-Long; Zhang, Ji-Wen; Wu, Quan-Xiang

    2016-03-23

    The fungus Clonostachys rosea is widely distributed all over the world. The destructive force of this fungus, as a biological control agent, is very strong to lots of plant pathogenic fungi. As part of the ongoing search for antibiotics from fungi obtained from soil samples, the secondary metabolites of C. rosea YRS-06 were investigated. Through efficient bioassay-guided isolation, three new bisorbicillinoids possessing open-ended cage structures, tetrahydrotrichodimer ether (1) and dihydrotrichodimer ether A and B (2 and 3), and 12 known compounds were obtained. Their structures were determined via extensive NMR, HR-ESI-MS, and CD spectroscopic analyses and X-ray diffraction data. Compounds 1-3 are rare bisorbicillinoids with a γ-pyrone moiety. The biological properties of 1-15 were evaluated against six different Gram-positive and Gram-negative bacteria. Bisorbicillinoids, 2-5, and TMC-151 C and E, 14 and 15, showed potent antibacterial activity.

  11. A numerical study of combined use of two biocontrol agents with different biocontrol mechanisms in controlling foliar pathogens.

    PubMed

    Xu, X-M; Jeffries, P; Pautasso, M; Jeger, M J

    2011-09-01

    Effective use of biocontrol agents is an important component of sustainable agriculture. A previous numerical study of a generic model showed that biocontrol efficacy was greatest for a single biocontrol agent (BCA) combining competition with mycoparasitism or antibiosis. This study uses the same mathematical model to investigate whether the biocontrol efficacy of combined use of two BCAs with different biocontrol mechanisms is greater than that of a single BCA with either or both of the two mechanisms, assuming that two BCAs occupy the same host tissue as the pathogen. Within the parameter values considered, a BCA with two biocontrol mechanisms always outperformed the combined use of two BCAs with a single but different biocontrol mechanism. Similarly, combined use of two BCAs with a single but different biocontrol mechanism is shown to be far less effective than that of a single BCA with both mechanisms. Disease suppression from combined use of two BCAs was very similar to that achieved by the more efficacious one. As expected, a higher BCA introduction rate led to increased disease suppression. Incorporation of interactions between two BCAs did not greatly affect the disease dynamics except when a mycoparasitic and, to a lesser extent, an antibiotic-producing BCA was involved. Increasing the competitiveness of a mycoparasitic BCA over a BCA whose biocontrol mechanism is either competition or antibiosis may lead to improved biocontrol initially and reduced fluctuations in disease dynamics. The present study suggests that, under the model assumptions, combined use of two BCAs with different biocontrol mechanisms in most cases only results in control efficacies similar to using the more efficacious one alone. These predictions are consistent with published experimental results, suggesting that combined use of BCAs should not be recommended without clear understanding of their main biocontrol mechanisms and relative competitiveness, and experimental evaluation.

  12. [Colonization characteristics of endophytic bacteria NJ13 in Panax ginseng and its biocontrol efficiency against Alternaria leaf spot of ginseng].

    PubMed

    Chen, Chang-Qing; Li, Tong; Li, Xin-Lian; Jiang, Yun; Tian, Lei; Xu, Peng

    2014-05-01

    To reveal the colonization characteristics in host of endophytic biocontrol bacteria NJ13 isolated from Panax ginseng, this study obtained the marked strain NJ13-R which was double antibiotic resistant to rifampicin and streptomycin through enhancing the method of inducing antibiotic. The colonization characteristics in ginseng and its biocontrol efficiency against Alternaria spot of ginseng in the field were studied. The results showed that the strain could colonize in root, stem and leaf of ginseng and the colonization amount was positive correlated with inoculation concentration. Meanwhile, the strain could infect and then transfer in different tissues of ginseng The colonization amount of strain in roots and leaves of ginseng increased first and then decreased. However, the tendency of colonization amount of strain in stems was ascend at first and then descend slowly, and was more than that in roots and leaves along with time, which had a preference to specific tissue of its host. In field experiment, the endophytic bacteria NJ13 was proved to be effective in controlling Alternaria leaf spot of ginseng. The biocontrol efficiency of fermentation broth at the concentration of 0.76 x 10(8) cfu x mL(-1) reached 75.62%, which was close to the controlling level (73.06%) of 0.67 mg x L(-1) 50% cyprodinil WG.

  13. Comparative Genomic Analysis of Pseudomonas chlororaphis PCL1606 Reveals New Insight into Antifungal Compounds Involved in Biocontrol.

    PubMed

    Calderón, Claudia E; Ramos, Cayo; de Vicente, Antonio; Cazorla, Francisco M

    2015-03-01

    Pseudomonas chlororaphis PCL1606 is a rhizobacterium that has biocontrol activity against many soilborne phytopathogenic fungi. The whole genome sequence of this strain was obtained using the Illumina Hiseq 2000 sequencing platform and was assembled using SOAP denovo software. The resulting 6.66-Mb complete sequence of the PCL1606 genome was further analyzed. A comparative genomic analysis using 10 plant-associated strains within the fluorescent Pseudomonas group, including the complete genome of P. chlororaphis PCL1606, revealed a diverse spectrum of traits involved in multitrophic interactions with plants and microbes as well as biological control. Phylogenetic analysis of these strains using eight housekeeping genes clearly placed strain PCL1606 into the P. chlororaphis group. The genome sequence of P. chlororaphis PCL1606 revealed the presence of sequences that were homologous to biosynthetic genes for the antifungal compounds 2-hexyl, 5-propyl resorcinol (HPR), hydrogen cyanide, and pyrrolnitrin; this is the first report of pyrrolnitrin encoding genes in this P. chlororaphis strain. Single-, double-, and triple-insertional mutants in the biosynthetic genes of each antifungal compound were used to test their roles in the production of these antifungal compounds and in antagonism and biocontrol of two fungal pathogens. The results confirmed the function of HPR in the antagonistic phenotype and in the biocontrol activity of P. chlororaphis PCL1606.

  14. Evaluation of economically feasible, natural plant extract-based microbiological media for producing biomass of the dry rot biocontrol strain Pseudomonas fluorescens P22Y05 in liquid culture.

    PubMed

    Khalil, Sadia; Ali, Tasneem Adam; Skory, Chris; Slininger, Patricia J; Schisler, David A

    2016-02-01

    The production of microbial biomass in liquid media often represents an indispensable step in the research and development of bacterial and fungal strains. Costs of commercially prepared nutrient media or purified media components, however, can represent a significant hurdle to conducting research in locations where obtaining these products is difficult. A less expensive option for providing components essential to microbial growth in liquid culture is the use of extracts of fresh or dried plant products obtained by using hot water extraction techniques. A total of 13 plant extract-based media were prepared from a variety of plant fruits, pods or seeds of plant species including Allium cepa (red onion bulb), Phaseolus vulgaris (green bean pods), and Lens culinaris (lentil seeds). In shake flask tests, cell production by potato dry rot antagonist Pseudomonas fluorescens P22Y05 in plant extract-based media was generally statistically indistinguishable from that in commercially produced tryptic soy broth and nutrient broth as measured by optical density and colony forming units/ml produced (P ≤ 0.05, Fisher's protected LSD). The efficacy of biomass produced in the best plant extract-based media or commercial media was equivalent in reducing Fusarium dry rot by 50-96% compared to controls. In studies using a high-throughput microbioreactor, logarithmic growth of P22Y05 in plant extract-based media initiated in 3-5 h in most cases but specific growth rate and the time of maximum OD varied as did the maximum pH obtained in media. Nutrient analysis of selected media before and after cell growth indicated that nitrogen in the form of NH4 accumulated in culture supernatants, possibly due to unbalanced growth conditions brought on by a scarcity of simple sugars in the media tested. The potential of plant extract-based media to economically produce biomass of microbes active in reducing plant disease is considerable and deserves further research.

  15. Metabolism of zearalenone by genetically modified organisms expressing the detoxification gene from Clonostachys rosea.

    PubMed

    Takahashi-Ando, Naoko; Ohsato, Shuichi; Shibata, Takehiko; Hamamoto, Hiroshi; Yamaguchi, Isamu; Kimura, Makoto

    2004-06-01

    Zearalenone (ZEN) is converted to a nontoxic product by a lactonohydololase encoded by zhd101. An enhanced green fluorescent protein (EGFP) gene was fused to zhd101 (i.e., egfp::zhd101) and expressed in Escherichia coli. Both recombinant ZHD101 and EGFP::ZHD101 were purified to homogeneity and characterized. Maximal activity of ZHD101 toward ZEN was measured at approximately 37 to 45 degrees C and pH 10.5 (k(cat) at 30 degrees C, 0.51 s(-1)). The enzyme was irreversibly inactivated at pH values below 4.5 or by treatment with serine protease inhibitors. ZHD101 was also active against five ZEN cognates, although the efficiencies were generally low; e.g., the k(cat) was highest with zearalanone (1.5 s(-1)) and lowest with beta-zearalenol (0.075 s(-1)). EGFP::ZHD101 had properties similar to those of the individual proteins with regard to the EGFP fluorescence and lactonohydrolase activity. Fortuitously, EGFP::ZHD101 exhibited a good correlation between the fluorescence intensity and reaction velocity under various pH conditions. We therefore used egfp::zhd101 to visually monitor the lactonohydrolase activity in genetically modified organisms and evaluated the usefulness of zhd101 for in vivo detoxification of ZEN. While recombinant E. coli and transgenic rice calluses exhibited strong EGFP fluorescence and completely degraded ZEN in liquid media, recombinant Saccharomyces cerevisiae gave poor fluorescence and did not eliminate all the toxicity of the mycotoxin in the medium; i.e., the rest of ZEN was transformed into an unfavorable substrate, beta-zearalenol, by an as-yet-unidentified reductase and remained in the medium. Even so, as much as 75% of ZEN was detoxified by the yeast transformant, which is better than the detoxification system in which food-grade Lactobacillus strains are used (H. El-Nezami, N. Polychronaki, S. Salminen, and H. Mykkuäne, Appl. Environ. Microbiol. 68:3545-3549, 2002). An appropriate combination of a candidate host microbe and the codon

  16. Metabolism of Zearalenone by Genetically Modified Organisms Expressing the Detoxification Gene from Clonostachys rosea

    PubMed Central

    Takahashi-Ando, Naoko; Ohsato, Shuichi; Shibata, Takehiko; Hamamoto, Hiroshi; Yamaguchi, Isamu; Kimura, Makoto

    2004-01-01

    Zearalenone (ZEN) is converted to a nontoxic product by a lactonohydololase encoded by zhd101. An enhanced green fluorescent protein (EGFP) gene was fused to zhd101 (i.e., egfp::zhd101) and expressed in Escherichia coli. Both recombinant ZHD101 and EGFP::ZHD101 were purified to homogeneity and characterized. Maximal activity of ZHD101 toward ZEN was measured at approximately 37 to 45°C and pH 10.5 (kcat at 30°C, 0.51 s−1). The enzyme was irreversibly inactivated at pH values below 4.5 or by treatment with serine protease inhibitors. ZHD101 was also active against five ZEN cognates, although the efficiencies were generally low; e.g., the kcat was highest with zearalanone (1.5 s−1) and lowest with β-zearalenol (0.075 s−1). EGFP::ZHD101 had properties similar to those of the individual proteins with regard to the EGFP fluorescence and lactonohydrolase activity. Fortuitously, EGFP::ZHD101 exhibited a good correlation between the fluorescence intensity and reaction velocity under various pH conditions. We therefore used egfp::zhd101 to visually monitor the lactonohydrolase activity in genetically modified organisms and evaluated the usefulness of zhd101 for in vivo detoxification of ZEN. While recombinant E. coli and transgenic rice calluses exhibited strong EGFP fluorescence and completely degraded ZEN in liquid media, recombinant Saccharomyces cerevisiae gave poor fluorescence and did not eliminate all the toxicity of the mycotoxin in the medium; i.e., the rest of ZEN was transformed into an unfavorable substrate, β-zearalenol, by an as-yet-unidentified reductase and remained in the medium. Even so, as much as 75% of ZEN was detoxified by the yeast transformant, which is better than the detoxification system in which food-grade Lactobacillus strains are used (H. El-Nezami, N. Polychronaki, S. Salminen, and H. Mykkuäne, Appl. Environ. Microbiol. 68:3545-3549, 2002). An appropriate combination of a candidate host microbe and the codon-optimized synthetic gene

  17. Interaction of media on production and biocontrol efficacy of Pseudomonas fluorescens and Bacillus subtilis against grey mould of apple.

    PubMed

    Peighamy-Ashnaei, S; Sharifi-Tehrani, A; Ahmadzadeh, M; Behboudi, K

    2008-01-01

    The medium has a profound effect on biocontrol agents, including ability to grow and effectiveness in disease control. In this study, growth and antagonistic efficacy of strains P-5 and P-35 (P. fluorescens), B-3 and B-16 (B. subtilis) were evaluated in combinations of two carbon (sucrose and molasses) and two nitrogen (urea and yeast extract) sources to optimize control of Botrytis cinerea on apple. All of the strains were grown in different liquid media (pH = 6.9) including: sucrose + yeast extract, molasses of sugar beet + yeast extract in 2:1 and 1:1 w/w ratios, molasses of sugar beet + urea, molasses, malt extract and nutrient broth. Apples (Golden Delicious) were inoculated by a 25-microl suspension of 10(6) spores of B. cinerea per ml, wounding each fruit (in two sites separately). Then a 25-microl suspension of each strain, containing 2 x 10(8) cfu ml(-1) grown in each of the above culture media, was applied to each wound. Results indicated that Molasses + Yeast extract (1:1 w/w) medium supported rapid growth in all of the strains. The final growth of B. subtilis B-16 in Molasses + Yeast extract (1:1 w/w) medium was 5 x 10(9) cfu ml(-1). After ten days, all of the strains significantly inhibited pathogenicity of B. cinerea on apples. The biocontrol efficacy of B. subtilis B-3 in Molasses + Yeast extract (1:1 w/w) medium reduced the severity of grey mould from 100% (inoculated control) to less than 26.9%. After 20 days, Strain B-3 showed a considerable biocontrol efficacy in Molasses medium and reduced the severity of grey mould from 100% (inoculated control) to less than 38.2%. The results obtained in this study could be used to provide a reliable basis for the increase of population of biocontrol agents in fermentation process.

  18. Phylogeny of certain biocontrol agents with special reference to nematophagous fungi based on RAPd.

    PubMed

    Jarullah, B M S; Subramanian, R B; Jummanah, M S J

    2005-01-01

    A number of phylogenetic studies have been carried out on biocontrol agents having similar biological control activity. However, no work has been carried out to determine the phylogenetic relationship amongst various groups of biological control agents with varied biocontrol properties. Our aim was to derive a phylogenetic relationship between diverse biocontrol agents belonging to the deuteromycetes and determine its correlation with their spore morphology and their biocontrol activity. RAPD was used to assess genomic variability in fungi used as biological control agents which included ten isolates of nematophagous fungi such as Arthrobotrys sp., Duddingtonia sp., Paecilomyces sp. and Verticillium sp., along with two isolates of fungal biocontrol agents such as Trichoderma sp. and two isolates of entomopathogenic fungi including Beauveria sp. A plant pathogenic fungus, Verticillium alboatrum was also included to increase the diversity of Deuteromycetes used. A similarity matrix was created using Jaccard's similarity coefficient & clustering was done using unweighted pair group arithmetic mean method (UPGMA). The final dendogram was created using a combination of two programs, Freetree and TreeExplorer. The phylogenetic tree constructed from the RAPD data showed marked genetic variability among different strains of the same species. The spore morphologies of all these fungi were also studied. The phylogenetic pattern could be correlated with the conidial and conidiophore morphology, a criterion commonly used for the classification of fungi in general and Deuteromycetes in particular. Interestingly, the inferred phylogeny showed no significant grouping based on either their biological control properties or the trapping structures amongst the nematophagous fungi as reported earlier by other workers. The phylogenetic pattern was also similar to the tree obtained by comparing the 18S rRNA sequences from the database. The result clearly indicates that the classical

  19. Importance of prumycin produced by Bacillus amyloliquefaciens SD-32 in biocontrol against cucumber powdery mildew disease.

    PubMed

    Tanaka, Keijitsu; Fukuda, Mutsumi; Amaki, Yusuke; Sakaguchi, Takatoshi; Inai, Koji; Ishihara, Atsushi; Nakajima, Hiromitsu

    2017-05-30

    Powdery mildew disease of cucurbits is caused mainly by Podosphaera fusca, which is one of the most important limiting factors in cucurbit production worldwide. Previously we reported that Bacillus amyloliquefaciens biocontrol strain SD-32 produces C17 bacillomycin D and [Ile]surfactin, and that these metabolites play important roles in SD-32's biocontrol over cucumber gray mold disease. Our further investigation demonstrated that the culture broth and its supernatant suppressed cucumber powdery mildew disease in greenhouse experiments. However, the active principle(s) remained unknown. The active compound was isolated from the culture supernatant after anti-powdery mildew disease activity-guided purification and identified as prumycin. Prumycin significantly suppressed the disease, whereas bacillomycin D and [Ile]surfactin did not. Prumycin did not induce the expression of plant defense genes (PR1a and VSP1), suggesting that it does not act via plant defense response. Light microscopic observations of prumycin-treated cucumber cotyledon suggested that prumycin inhibits the conidial germination of P. fusca. This study demonstrates that prumycin is a major factor in SD-32's suppression of cucumber powdery mildew disease. Our findings shed light for the first time on prumycin's role in biocontrol by Bacillus against this disease. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  20. Bacteriophage-Based Bacterial Wilt Biocontrol for an Environmentally Sustainable Agriculture.

    PubMed

    Álvarez, Belén; Biosca, Elena G

    2017-01-01

    Bacterial wilt diseases caused by Ralstonia solanacearum, R. pseudosolanacearum, and R. syzygii subsp. indonesiensis (former R. solanacearum species complex) are among the most important plant diseases worldwide, severely affecting a high number of crops and ornamentals. Difficulties of bacterial wilt control by non-biological methods are related to effectiveness, bacterial resistance and environmental impact. Alternatively, a great many biocontrol strategies have been carried out, with the advantage of being environmentally friendly. Advances in bacterial wilt biocontrol include an increasing interest in bacteriophage-based treatments as a promising re-emerging strategy. Bacteriophages against the bacterial wilt pathogens have been described with either lytic or lysogenic effect but, they were proved to be active against strains belonging to R. pseudosolanacearum and/or R. syzygii subsp. indonesiensis, not to the present R. solanacearum species, and only two of them demonstrated successful biocontrol potential in planta. Despite the publication of three patents on the topic, until now no bacteriophage-based product is commercially available. Therefore, there is still much to be done to incorporate valid bacteriophages in an integrated management program to effectively fight bacterial wilt in the field.

  1. Bacteriophage-Based Bacterial Wilt Biocontrol for an Environmentally Sustainable Agriculture

    PubMed Central

    Álvarez, Belén; Biosca, Elena G.

    2017-01-01

    Bacterial wilt diseases caused by Ralstonia solanacearum, R. pseudosolanacearum, and R. syzygii subsp. indonesiensis (former R. solanacearum species complex) are among the most important plant diseases worldwide, severely affecting a high number of crops and ornamentals. Difficulties of bacterial wilt control by non-biological methods are related to effectiveness, bacterial resistance and environmental impact. Alternatively, a great many biocontrol strategies have been carried out, with the advantage of being environmentally friendly. Advances in bacterial wilt biocontrol include an increasing interest in bacteriophage-based treatments as a promising re-emerging strategy. Bacteriophages against the bacterial wilt pathogens have been described with either lytic or lysogenic effect but, they were proved to be active against strains belonging to R. pseudosolanacearum and/or R. syzygii subsp. indonesiensis, not to the present R. solanacearum species, and only two of them demonstrated successful biocontrol potential in planta. Despite the publication of three patents on the topic, until now no bacteriophage-based product is commercially available. Therefore, there is still much to be done to incorporate valid bacteriophages in an integrated management program to effectively fight bacterial wilt in the field. PMID:28769942

  2. Response to Thomas et al.: Biocontrol and indirect effects

    Treesearch

    Dean E. Pearson; Ragan M. Callaway

    2004-01-01

    In a recent TREE article [1], we identified three categories of unintended indirect effects that can arise from host-specific biological control agents: (i) ecological replacement; (ii) compensatory responses; and (iii) food-web interactions. Although our review focused on the biocontrol of plant pests, we suggested these concepts also apply to the biocontrol...

  3. Recent taxonomy changes and their impact on biocontrol agents

    USDA-ARS?s Scientific Manuscript database

    The revolution in DNA sequencing technology has led to and improved understanding of genetics and taxonomy of biocontrol agents. Our lab recently reported the genomes of some important Bacillus bacterial biocontrol agents, which in turn resulted in a change of taxonomy for these commercially importa...

  4. Selection and evaluation of micro-organisms for biocontrol of Verticillium dahliae in olive.

    PubMed

    Varo, A; Raya-Ortega, M C; Trapero, A

    2016-09-01

    To identify potential biological control agents against Verticillium wilt in olive through a mass screening approach. A total of 47 strains and nine mixtures of micro-organisms were evaluated against Verticillium dahliae in a three stage screening: (i) in vitro, by the effect on the mycelial growth and spore germination of the pathogen; (ii) in natural infested soil, by the effect on the reduction of microsclerotia of the pathogen; (iii) in planta, by the effect on the infection of olive plants under controlled conditions. Various fungal and bacterial strains and mixtures inhibited the pathogen and showed consistent biocontrol activity against Verticillium wilt of olive. The screening has resulted in promising fungi and bacteria strains with antagonistic activity against Verticillium, such as two non-pathogenic Fusarium oxysporum, one Phoma sp., one Pseudomonas fluorescens and two mixtures of micro-organisms that may possess multiple modes of action. This study provides a practical basis for the potential use of selected strains as biocontrol agents for the protection of olive plants against V. dahliae infection. In addition, our study presented an effective method to evaluate antagonistic micro-organisms of V. dahliae in olive. © 2016 The Society for Applied Microbiology.

  5. Lactic acid bacteria from fresh fruit and vegetables as biocontrol agents of phytopathogenic bacteria and fungi.

    PubMed

    Trias, Rosalia; Bañeras, Lluís; Montesinos, Emilio; Badosa, Esther

    2008-12-01

    This study evaluated the efficacy of lactic acid bacteria (LAB) isolated from fresh fruits and vegetables as biocontrol agents against the phytopathogenic and spoilage bacteria and fungi, Xanthomonas campestris, Erwinia carotovora, Penicillium expansum, Monilinia laxa, and Botrytis cinerea. The antagonistic activity of 496 LAB strains was tested in vitro and all tested microorganisms except P. expansum were inhibited by at least one isolate. The 496 isolates were also analyzed for the inhibition of P. expansum infection in wounds of Golden Delicious apples. Four strains (TC97, AC318, TM319, and FF441) reduced the fungal rot diameter of the apples by 20%; only Weissella cibaria strain TM128 decreased infection levels by 50%. Cell-free supernatants of selected antagonistic bacteria were studied to determine the nature of the antimicrobial compounds produced. Organic acids were the preferred mediators of inhibition but hydrogen peroxide was also detected when strains BC48, TM128, PM141 and FF441 were tested against E. carotovora. While previous reports of antifungal activity by LAB are scarce, our results support the potential of LAB as biocontrol agents against postharvest rot.

  6. Influence of Pythium oligandrum Biocontrol on Fungal and Oomycete Population Dynamics in the Rhizosphere▿

    PubMed Central

    Vallance, Jessica; Le Floch, Gaétan; Déniel, Franck; Barbier, Georges; Lévesque, C. André; Rey, Patrice

    2009-01-01

    Fungal and oomycete populations and their dynamics were investigated following the introduction of the biocontrol agent Pythium oligandrum into the rhizosphere of tomato plants grown in soilless culture. Three strains of P. oligandrum were selected on the basis of their ability to form oospores (resting structures) and to produce tryptamine (an auxin-like compound) and oligandrin (a glycoprotein elicitor). Real-time PCR and plate counting demonstrated the persistence of large amounts of the antagonistic oomycete in the rhizosphere throughout the cropping season (April to September). Inter-simple-sequence-repeat analysis of the P. oligandrum strains collected from root samples at the end of the cropping season showed that among the three strains used for inoculation, the one producing the smallest amount of oospores was detected at 90%. Single-strand conformational polymorphism analysis revealed increases in the number of members and the complexity of the fungal community over time. There were no significant differences between the microbial ecosystems inoculated with P. oligandrum and those that were not treated, except for a reduction of Pythium dissotocum (ubiquitous tomato root minor pathogen) populations in inoculated systems during the last 3 months of culture. These findings raise interesting issues concerning the use of P. oligandrum strains producing elicitor and auxin molecules for plant protection and the development of biocontrol. PMID:19447961

  7. Combinations of Biocontrol Agents for Management of Plant-Parasitic Nematodes and Soilborne Plant-Pathogenic Fungi

    PubMed Central

    Meyer, Susan L. F.; Roberts, Daniel P.

    2002-01-01

    Numerous microbes are antagonistic to plant-parasitic nematodes and soilborne plant-pathogenic fungi, but few of these organisms are commercially available for management of these pathogens. Inconsistent performance of applied biocontrol agents has proven to be a primary obstacle to the development of successful commercial products. One of the strategies for overcoming inconsistent performance is to combine the disease-suppressive activity of two (or more) beneficial microbes in a biocontrol preparation. Such combinations have potential for more extensive colonization of the rhizosphere, more consistent expression of beneficial traits under a broad range of soil conditions, and antagonism to a larger number of plant pests or pathogens than strains applied individually. Conversely, microbes applied in combination also may have antagonistic interactions with each other. Increased, decreased, and unaltered suppression of the target pathogen or pest has been observed when biocontrol microbes have been applied in combination. Unfortunately, the ecological basis for increased or decreased suppression has not been determined in many cases and needs further consideration. The complexity of interactions involved in the application of multiple organisms for biological control has slowed progress toward development of successful formulations. However, this approach has potential for overcoming some of the efficacy problems that occur with application of individual biocontrol agents. PMID:19265899

  8. The phosphotransferase system gene ptsI in the endophytic bacterium Bacillus cereus is required for biofilm formation, colonization, and biocontrol against wheat sharp eyespot.

    PubMed

    Xu, Yu-Bin; Chen, Mai; Zhang, Ying; Wang, Miao; Wang, Ying; Huang, Qiu-bin; Wang, Xue; Wang, Gang

    2014-05-01

    Natural resistance of wheat plants to wheat sharp eyespot is inadequate, and new strategies for controlling the disease are required. Biological control is an alternative and attractive way of reducing the use of chemicals in agriculture. In this study, we investigated the biocontrol properties of endophytic bacterium Bacillus cereus strain 0-9, which was isolated from the root systems of healthy wheat varieties. The phosphotransferase system is a major regulator of carbohydrate metabolism in bacteria. Enzyme I is one of the protein components of this system. Specific disruption and complementation of the enzyme I-coding gene ptsI from B. cereus was achieved through homologous recombination. Disruption of ptsI in B. cereus caused a 70% reduction in biofilm formation, a 30.4% decrease in biocontrol efficacy, and a 1000-fold reduction in colonization. The growth of ΔptsI mutant strain on G-tris synthetic medium containing glucose as the exclusive carbon source was also reduced. Wild-type properties could be restored to the ΔptsI mutant strain by ptsI complementation. These results suggested that ptsI may be one of the key genes involved in biofilm formation, colonization, and biocontrol of B. cereus and that B. cereus wild-type strain 0-9 may be an ideal biocontrol agent for controlling wheat sharp eyespot.

  9. Genotypic comparison of Pantoea agglomerans plant and clinical strains

    PubMed Central

    2009-01-01

    Background Pantoea agglomerans strains are among the most promising biocontrol agents for a variety of bacterial and fungal plant diseases, particularly fire blight of apple and pear. However, commercial registration of P. agglomerans biocontrol products is hampered because this species is currently listed as a biosafety level 2 (BL2) organism due to clinical reports as an opportunistic human pathogen. This study compares plant-origin and clinical strains in a search for discriminating genotypic/phenotypic markers using multi-locus phylogenetic analysis and fluorescent amplified fragment length polymorphisms (fAFLP) fingerprinting. Results Majority of the clinical isolates from culture collections were found to be improperly designated as P. agglomerans after sequence analysis. The frequent taxonomic rearrangements underwent by the Enterobacter agglomerans/Erwinia herbicola complex may be a major problem in assessing clinical associations within P. agglomerans. In the P. agglomerans sensu stricto (in the stricter sense) group, there was no discrete clustering of clinical/biocontrol strains and no marker was identified that was uniquely associated to clinical strains. A putative biocontrol-specific fAFLP marker was identified only in biocontrol strains. The partial ORF located in this band corresponded to an ABC transporter that was found in all P. agglomerans strains. Conclusion Taxonomic mischaracterization was identified as a major problem with P. agglomerans, and current techniques removed a majority of clinical strains from this species. Although clear discrimination between P. agglomerans plant and clinical strains was not obtained with phylogenetic analysis, a single marker characteristic of biocontrol strains was identified which may be of use in strain biosafety determinations. In addition, the lack of Koch's postulate fulfilment, rare retention of clinical strains for subsequent confirmation, and the polymicrobial nature of P. agglomerans clinical reports

  10. Endemic plants harbour specific Trichoderma communities with an exceptional potential for biocontrol of phytopathogens.

    PubMed

    Zachow, Christin; Berg, Christian; Müller, Henry; Monk, Jana; Berg, Gabriele

    2016-10-10

    Trichoderma strains exhibit enormous potential for applications in biotechnology, in particular as biocontrol agents against pathogens. However, little is known about the diversity of plant-associated Trichoderma communities at a global scale and their antagonistic spectrum. In order to gather information about structure and function, we compared Trichoderma biomes of endemic (Aeonium, Diospyros, Hebe, Rhododendron) and cosmopolitan plants (Zea mays) in a global study encompassing the area Northwest Africa to New Zealand via the European Alps and Madagascar. At the quantitative level we found no differences between cosmopolitan and endemic plants. Statistically significant differences were detected at the qualitative level: Trichoderma populations of endemic plants were highly specific and diverse with hot spots appearing in Madagascar and New Zealand. By contrast, maize plants from all sites shared the majority of Trichoderma species (65.5%). Interestingly, the high above ground biodiversity in ecosystems containing endemic plants was confirmed by a high below ground Trichoderma diversity. Despite the differences, we found a global Trichoderma core community shared by all analysed plants, which was dominated by T. koningii and T. koningiopsis. Amplicon-based network analyses revealed a high similarity between maize Trichoderma grown world-wide and distinct populations of endemic plants. Furthermore, Trichoderma strains from endemic plants showed a higher antagonistic activity against fungal pathogens compared to maize-associated strains. Our results showed that endemic plants are associated with a specific Trichoderma microbiome which possesses a high antagonistic activity indicating that it has potential to be used for biocontrol purposes.

  11. Diversity, Biocontrol, and Plant Growth Promoting Abilities of Xylem Residing Bacteria from Solanaceous Crops

    PubMed Central

    Achari, Gauri A.

    2014-01-01

    Eggplant (Solanum melongena L.) is one of the solanaceous crops of economic and cultural importance and is widely cultivated in the state of Goa, India. Eggplant cultivation is severely affected by bacterial wilt caused by Ralstonia solanacearum that colonizes the xylem tissue. In this study, 167 bacteria were isolated from the xylem of healthy eggplant, chilli, and Solanum torvum Sw. by vacuum infiltration and maceration. Amplified rDNA restriction analysis (ARDRA) grouped these xylem residing bacteria (XRB) into 38 haplotypes. Twenty-eight strains inhibited growth of R. solanacearum and produced volatile and diffusible antagonistic compounds and plant growth promoting substances in vitro. Antagonistic strains XB86, XB169, XB177, and XB200 recorded a biocontrol efficacy greater than 85% against BW and exhibited 12%–22 % increase in shoot length in eggplant in the greenhouse screening. 16S rRNA based identification revealed the presence of 23 different bacterial genera. XRB with high biocontrol and plant growth promoting activities were identified as strains of Staphylococcus sp., Bacillus sp., Streptomyces sp., Enterobacter sp., and Agrobacterium sp. This study is the first report on identity of bacteria from the xylem of solanaceous crops having traits useful in cultivation of eggplant. PMID:24963298

  12. 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.

  13. Biocontrol: Bacillus penetrans and Related Parasites of Nematodes

    PubMed Central

    Sayre, R. M.

    1980-01-01

    Bacillus penetrans Mankau, 1975, previously described as Duboscqia penetrans Thorne 1940, is a candidate agent for biocontrol of nematodes. This review considers the life stages of this bacterium: vegetative growth phase, colony fragmentation, sporogenesis, soil phase, spore attachment, and penetration into larvae of root-knot nematodes. The morphology of the microthallus colonies and the unusual external features of the spore are discussed. Taxonomic affinities with the actinomycetes, particularly with the genus Pasteuria, are considered. Also discussed are other soil bacterial species that are potential biocontrol agents. Products of their bacterial fermentation in soil are toxic to nematodes, making them effective biocontrol agents. PMID:19300701

  14. Nonribosomal peptides, key biocontrol components for Pseudomonas fluorescens In5, isolated from a Greenlandic suppressive soil.

    PubMed

    Michelsen, Charlotte F; Watrous, Jeramie; Glaring, Mikkel A; Kersten, Roland; Koyama, Nobuhiro; Dorrestein, Pieter C; Stougaard, Peter

    2015-03-17

    Potatoes are cultivated in southwest Greenland without the use of pesticides and with limited crop rotation. Despite the fact that plant-pathogenic fungi are present, no severe-disease outbreaks have yet been observed. In this report, we document that a potato soil at Inneruulalik in southern Greenland is suppressive against Rhizoctonia solani Ag3 and uncover the suppressive antifungal mechanism of a highly potent biocontrol bacterium, Pseudomonas fluorescens In5, isolated from the suppressive potato soil. A combination of molecular genetics, genomics, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) imaging mass spectrometry (IMS) revealed an antifungal genomic island in P. fluorescens In5 encoding two nonribosomal peptides, nunamycin and nunapeptin, which are key components for the biocontrol activity by strain In5 in vitro and in soil microcosm experiments. Furthermore, complex microbial behaviors were highlighted. Whereas nunamycin was demonstrated to inhibit the mycelial growth of R. solani Ag3, but not that of Pythium aphanidermatum, nunapeptin instead inhibited P. aphanidermatum but not R. solani Ag3. Moreover, the synthesis of nunamycin by P. fluorescens In5 was inhibited in the presence of P. aphanidermatum. Further characterization of the two peptides revealed nunamycin to be a monochlorinated 9-amino-acid cyclic lipopeptide with similarity to members of the syringomycin group, whereas nunapeptin was a 22-amino-acid cyclic lipopeptide with similarity to corpeptin and syringopeptin. Crop rotation and systematic pest management are used to only a limited extent in Greenlandic potato farming. Nonetheless, although plant-pathogenic fungi are present in the soil, the farmers do not experience major plant disease outbreaks. Here, we show that a Greenlandic potato soil is suppressive against Rhizoctonia solani, and we unravel the key biocontrol components for Pseudomonas fluorescens In5, one of the potent biocontrol bacteria

  15. Rhizobia: a potential biocontrol agent for soilborne fungal pathogens.

    PubMed

    Das, Krishnashis; Prasanna, Radha; Saxena, Anil Kumar

    2017-03-12

    Rhizobia are a group of organisms that are well known for their ability to colonize root surfaces and form symbiotic associations with legume plants. They not only play a major role in biological nitrogen fixation but also improve plant growth and reduce disease incidence in various crops. Rhizobia are known to control the growth of many soilborne plant pathogenic fungi belonging to different genera like Fusarium, Rhizoctonia, Sclerotium, and Macrophomina. Antagonistic activity of rhizobia is mainly attributed to production of antibiotics, hydrocyanic acid (HCN), mycolytic enzymes, and siderophore under iron limiting conditions. Rhizobia are also reported to induce systemic resistance and enhance expression of plant defense-related genes, which effectively immunize the plants against pathogens. Seed bacterization with appropriate rhizobial strain leads to elicitation and accumulation of phenolic compounds, isoflavonoid phytoalexins, and activation of enzymes like L-phenylalanine ammonia lyase (PAL), chalcone synthase (CHS), peroxidase (POX), polyphenol oxidase (PPO), and others involved in phenylpropanoid and isoflavonoid pathways. Development of Rhizobium inoculants with dual attributes of nitrogen fixation and antagonism against phytopathogens can contribute to increased plant growth and productivity. This compilation aims to bring together the available information on the biocontrol facet of rhizobia and identify research gaps and effective strategies for future research in this area.

  16. Selection and characterization of Argentine isolates of Trichoderma harzianum for effective biocontrol of Septoria leaf blotch of wheat.

    PubMed

    Stocco, Marina C; Mónaco, Cecilia I; Abramoff, Cecilia; Lampugnani, Gladys; Salerno, Graciela; Kripelz, Natalia; Cordo, Cristina A; Consolo, Verónica F

    2016-03-01

    Species of the genus Trichoderma are economically important as biocontrol agents, serving as a potential alternative to chemical control. The applicability of Trichoderma isolates to different ecozones will depend on the behavior of the strains selected from each zone. The present study was undertaken to isolate biocontrol populations of Trichoderma spp. from the Argentine wheat regions and to select and characterize the best strains of Trichoderma harzianum by means of molecular techniques. A total of 84 out of the 240 strains of Trichoderma were able to reduce the disease severity of the leaf blotch of wheat. Thirty-seven strains were selected for the reduction equal to or greater than 50% of the severity, compared with the control. The percentage values of reduction of the pycnidial coverage ranged between 45 and 80%. The same last strains were confirmed as T. harzianum by polymerase chain reaction amplification of internal transcribed spacers, followed by sequencing. Inter-simple sequence repeat was used to examine the genetic variability among isolates. This resulted in a total of 132 bands. Further numerical analysis revealed 19 haplotypes, grouped in three clusters (I, II, III). Shared strains, with different geographical origins and isolated in different years, were observed within each cluster. The origin of the isolates and the genetic group were partially related. All isolates from Paraná were in cluster I, all isolates from Lobería were in cluster II, and all isolates from Pergamino and Santa Fe were in cluster III. Our results suggest that the 37 native strains of T. harzianum are important in biocontrol programs and could be advantageous for the preparation of biopesticides adapted to the agroecological conditions of wheat culture.

  17. Biological control of soilborne diseases in organic potato production using hypovirulent strains of Rhizoctonia solani

    USDA-ARS?s Scientific Manuscript database

    Soilborne diseases are persistent problems in potato production and alternative management practices are needed, particularly in organic production, where control options are limited. Selected biocontrol organisms, including two naturally-occurring hypovirulent strains of Rhizoctonia solani (Rhs1a1 ...

  18. Biocontrol interventions for inactivation of foodborne pathogens on produce

    USDA-ARS?s Scientific Manuscript database

    Post-harvest interventions for control of foodborne pathogens on minimally processed foods are crucial for food safety. Biocontrol interventions have the primary objective of developing novel antagonists in combinations with physical and chemical interventions to inactivate pathogenic microbes. Ther...

  19. Biocontrol potential of Halotolerant bacterial chitinase from high yielding novel Bacillus Pumilus MCB-7 autochthonous to mangrove ecosystem.

    PubMed

    Rishad, K S; Rebello, Sharrel; Shabanamol, P S; Jisha, M S

    2017-04-01

    The multifaceted role of chitinase in medicine, agriculture, environmental remediation and various other industries greatly demands the isolation of high yielding chitinase producing microorganisms with improved properties. The current study aimed to investigate the isolation, characterization and biocontrol prospective of chitinase producing bacterial strains autochthonous to the extreme conditions of mangrove ecosystems. Among the 51 bacterial isolates screened, Bacillus pumilus MCB-7 with highest chitinase production potential was identified and confirmed by 16S rDNA typing. Chitinase enzyme of MCB-7 was purified; the chitin degradation was evaluated by SEM and LC-MS. Unlike previously reported B.pumilus isolates, MCB-7 exhibited highest chitinase activity of 3.36U/mL, active even at high salt concentrations and temperature up to 60°C. The crude as well as purified enzyme showed significant antimycotic activity against agricultural pathogens such as Aspergillus flavus, Aspergillus niger, Aspergillus fumigatus, Ceratorhiza hydrophila and Fusarium oxysporum. The enzyme also exhibited biopesticidal role against larvae of Scirpophaga incertulas (Walker). [Lep.: Pyralidae], a serious agricultural pest of rice. The high chitinolytic and antimycotic potential of MCB-7 increases the prospects of the isolate as an excellent biocontrol agent. To the best of our knowledge, this is the first report of high chitinase yielding Bacillus pumilus strain from mangrove ecosystem with a biocontrol role against phytopathogenic fungi and insect larval pests. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Bacteriophage cocktail for biocontrol of Salmonella in dried pet food.

    PubMed

    Heyse, Serena; Hanna, Leigh Farris; Woolston, Joelle; Sulakvelidze, Alexander; Charbonneau, Duane

    2015-01-01

    Human salmonellosis has been associated with contaminated pet foods and treats. Therefore, there is interest in identifying novel approaches for reducing the risk of Salmonella contamination within pet food manufacturing environments. The use of lytic bacteriophages shows promise as a safe and effective way to mitigate Salmonella contamination in various food products. Bacteriophages are safe, natural, highly targeted antibacterial agents that specifically kill bacteria and can be targeted to kill food pathogens without affecting other microbiota. In this study, we show that a cocktail containing six bacteriophages had a broadspectrum activity in vitro against a library of 930 Salmonella enterica strains representing 44 known serovars. The cocktail was effective against 95% of the strains in this tested library. In liquid culture dose-ranging experiments, bacteriophage cocktail concentrations of ≥10(8) PFU/ml inactivated more than 90% of the Salmonella population (10(1) to 10(3) CFU/ml). Dried pet food inoculated with a mixture containing equal proportions of Salmonella serovars Enteritidis (ATCC 4931), Montevideo (ATCC 8387), Senftenberg (ATCC 8400), and Typhimurium (ATCC 13311) and then surface treated with the six-bacteriophage cocktail (≥2.5 ± 1.5 × 10(6) PFU/g) achieved a greater than 1-log (P < 0.001) reduction compared with the phosphate-buffered saline-treated control in measured viable Salmonella within 60 min. Moreover, this bacteriophage cocktail reduced natural contamination in samples taken from an undistributed lot of commercial dried dog food that tested positive for Salmonella. Our results indicate that bacteriophage biocontrol of S. enterica in dried pet food is technically feasible.

  1. Bio-Control of Salmonella Enteritidis in Foods Using Bacteriophages.

    PubMed

    Bao, Hongduo; Zhang, Pengyu; Zhang, Hui; Zhou, Yan; Zhang, Lili; Wang, Ran

    2015-08-24

    Two lytic phages, vB_SenM-PA13076 (PA13076) and vB_SenM-PC2184 (PC2184), were isolated from chicken sewage and characterized with host strains Salmonella Enteritidis (SE) ATCC13076 and CVCC2184, respectively. Transmission electron microscopy revealed that they belonged to the family Myoviridae. The lytic abilities of these two phages in liquid culture showed 104 multiplicity of infection (MOI) was the best in inhibiting bacteria, with PC2184 exhibiting more activity than PA13076. The two phages exhibited broad host range within the genus Salmonella. Phage PA13076 and PC2184 had a lytic effect on 222 (71.4%) and 298 (95.8%) of the 311 epidemic Salmonella isolates, respectively. We tested the effectiveness of phage PA13076 and PC2184 as well as a cocktail combination of both in three different foods (chicken breast, pasteurized whole milk and Chinese cabbage) contaminated with SE. Samples were spiked with 1 × 10(4) CFU individual SE or a mixture of strains (ATCC13076 and CVCC2184), then treated with 1 × 10(8) PFU individual phage or a two phage cocktail, and incubated at 4 °C or 25 °C for 5 h. In general, the inhibitory effect of phage and phage cocktail was better at 4 °C than that at 25 °C, whereas the opposite result was observed in Chinese cabbage, and phage cocktail was better than either single phage. A significant reduction in bacterial numbers (1.5-4 log CFU/sample, p < 0.05) was observed in all tested foods. The two phages on the three food samples were relatively stable, especially at 4 ºC, with the phages exhibiting the greatest stability in milk. Our research shows that our phages have potential effectiveness as a bio-control agent of Salmonella in foods.

  2. Antagonistic and Biocontrol Potential of Trichoderma asperellum ZJSX5003 Against the Maize Stalk Rot Pathogen Fusarium graminearum.

    PubMed

    Li, Yaqian; Sun, Ruiyan; Yu, Jia; Saravanakumar, Kandasamy; Chen, Jie

    2016-09-01

    The efficacy of seven strains of Trichoderma asperellum collected from the fields in Southern China was assessed against Fusarium graminearum (FG) the causal agent of corn stalk rot of maize were in vitro for their antagonistic properties followed by statistical model of principal compound analysis to identify the beneficial antagonist T. asperellum strain. The key factors of antagonist activity were attributed to a total of 13 factors including cell wall degrading enzymes (chitnase, protease and β-glucanases), secondary metabolites and peptaibols and these were analyzed from eight strains of Trichoderma. A linear regression model demonstrated that interaction of enzymes and secondary metabolites of T. asperellum strain ZJSX5003 enhanced the antagonist activity against FG. Further, this strain displayed a disease reduction of 71 % in maize plants inoculated with FG compared to negative control. Pointing out that the T. asperellum strain ZJSX5003 is a potential source for the development of a biocontrol agent against corn stalk rot.

  3. Bacteriophages as biocontrol agents in food.

    PubMed

    Hudson, J A; Billington, C; Carey-Smith, G; Greening, G

    2005-02-01

    Bacteriophages possess attributes that appear to be attractive to those searching for novel ways to control foodborne pathogens and spoilage organisms. These phages have a history of safe use, can be highly host specific, and replicate in the presence of a host. Campylobacter, Salmonella, and Listeria monocytogenes and various spoilage organisms have responded to phage control on some foods. However, the use of phages as biocontrol agents is complicated by factors such as an apparent requirement for a threshold level of host before replication can proceed and by suboptimal performance, at best, at temperatures beneath the optimum for the host. This review is a summary of the information on these issues and includes brief descriptions of alternative phage-based strategies for control of foodborne pathogens.

  4. Protozoa Drive the Dynamics of Culturable Biocontrol Bacterial Communities.

    PubMed

    Müller, Maren Stella; Scheu, Stefan; Jousset, Alexandre

    2013-01-01

    Some soil bacteria protect plants against soil-borne diseases by producing toxic secondary metabolites. Such beneficial biocontrol bacteria can be used in agricultural systems as alternative to agrochemicals. The broad spectrum toxins responsible for plant protection also inhibit predation by protozoa and nematodes, the main consumers of bacteria in soil. Therefore, predation pressure may favour biocontrol bacteria and contribute to plant health. We analyzed the effect of Acanthamoeba castellanii on semi-natural soil bacterial communities in a microcosm experiment. We determined the frequency of culturable bacteria carrying genes responsible for the production of the antifungal compounds 2,4-diacetylphloroglucinol (DAPG), pyrrolnitrin (PRN) and hydrogen cyanide (HCN) in presence and absence of A. castellanii. We then measured if amoebae affected soil suppressiveness in a bioassay with sugar beet seedlings confronted to the fungal pathogen Rhizoctonia solani. Amoebae increased the frequency of both DAPG and HCN positive bacteria in later plant growth phases (2 and 3 weeks), as well as the average number of biocontrol genes per bacterium. The abundance of DAPG positive bacteria correlated with disease suppression, suggesting that their promotion by amoebae may enhance soil health. However, the net effect of amoebae on soil suppressiveness was neutral to slightly negative, possibly because amoebae slow down the establishment of biocontrol bacteria on the recently emerged seedlings used in the assay. The results indicate that microfaunal predators foster biocontrol bacterial communities. Understanding interactions between biocontrol bacteria and their predators may thus help developing environmentally friendly management practices of agricultural systems.

  5. Protozoa Drive the Dynamics of Culturable Biocontrol Bacterial Communities

    PubMed Central

    Müller, Maren Stella; Scheu, Stefan; Jousset, Alexandre

    2013-01-01

    Some soil bacteria protect plants against soil-borne diseases by producing toxic secondary metabolites. Such beneficial biocontrol bacteria can be used in agricultural systems as alternative to agrochemicals. The broad spectrum toxins responsible for plant protection also inhibit predation by protozoa and nematodes, the main consumers of bacteria in soil. Therefore, predation pressure may favour biocontrol bacteria and contribute to plant health. We analyzed the effect of Acanthamoeba castellanii on semi-natural soil bacterial communities in a microcosm experiment. We determined the frequency of culturable bacteria carrying genes responsible for the production of the antifungal compounds 2,4-diacetylphloroglucinol (DAPG), pyrrolnitrin (PRN) and hydrogen cyanide (HCN) in presence and absence of A. castellanii. We then measured if amoebae affected soil suppressiveness in a bioassay with sugar beet seedlings confronted to the fungal pathogen Rhizoctonia solani. Amoebae increased the frequency of both DAPG and HCN positive bacteria in later plant growth phases (2 and 3 weeks), as well as the average number of biocontrol genes per bacterium. The abundance of DAPG positive bacteria correlated with disease suppression, suggesting that their promotion by amoebae may enhance soil health. However, the net effect of amoebae on soil suppressiveness was neutral to slightly negative, possibly because amoebae slow down the establishment of biocontrol bacteria on the recently emerged seedlings used in the assay. The results indicate that microfaunal predators foster biocontrol bacterial communities. Understanding interactions between biocontrol bacteria and their predators may thus help developing environmentally friendly management practices of agricultural systems. PMID:23840423

  6. Nonribosomal peptides and polyketides of Burkholderia: new compounds potentially implicated in biocontrol and pharmaceuticals.

    PubMed

    Esmaeel, Qassim; Pupin, Maude; Jacques, Philippe; Leclère, Valérie

    2017-05-25

    Bacteria belonging to the genus Burkholderia live in various ecological niches and present a significant role in the environments through the excretion of a wide variety of secondary metabolites including modular nonribosomal peptides (NRPs) and polyketides (PKs). These metabolites represent a widely distributed biomedically and biocontrol important class of natural products including antibiotics, siderophores, and anticancers as well as biopesticides that are considered as a novel source that can be used to defend ecological niche from competitors and to promote plant growth. The aim of this review is to present all NRPs produced or potentially produced by strains of Burkholderia, as NRPs represent a major source of active compounds implicated in biocontrol. The review is a compilation of results from a large screening we have performed on 48 complete sequenced genomes available in NCBI to identify NRPS gene clusters, and data found in the literature mainly because some interesting compounds are produced by strains not yet sequenced. In addition to NRPs, hybrids NRPs/PKs are also included. Specific features about biosynthetic gene clusters and structures of the modular enzymes responsible for the synthesis, the biological activities, and the potential uses in agriculture and pharmaceutical of NRPs and hybrids NRPs/PKs will also be discussed.

  7. Carbon requirements of some nematophagous, entomopathogenic and mycoparasitic hyphomycetes as fungal biocontrol agents.

    PubMed

    Sun, ManHong; Liu, XingZhong

    2006-05-01

    Thirty-three carbon sources were evaluated for their effects on spore germination, hyphal growth and sporulation of 11 fungal biocontrol agents, i.e. the nematophagous fungi Paecilomyces lilacinus, Pochonia chlamydosporia, Hirsutella rhossiliensis, H. minnesotensis and Arkansas Fungus 18, the entomopathogenic fungi Lecanicillium lecanii, Beauveria bassiana and Metarhizium anisopliae, and the mycoparasitic fungus Trichoderma viride. Variations in carbon requirements were found among the fungal species or strains tested. All strains studied except for T. viride grew on most carbon sources, although B. bassiana had more fastidious requirements for spore germination. Monosaccharides and disaccharides were suitable for fungal growth. For most isolates, D-glucose, D-mannose, sucrose and trehalose were superior to pectin and soluble starch among the polysaccharides and lactic acid among the organic acids. Both ethanol and methanol could accelerate growth of most isolates but not biomass. D-mannose, D-fructose and D-xylose were excellent carbon sources for sporulation, while D-glucose, sucrose, cellobiose, trehalose, chitin, dextrin, gelatin and lactic acid were better for some isolates. Neither sorbic acid nor linoleic acid could be utilized as a single carbon source. These findings provided a better understanding of the nutritional requirements of different fungal biocontrol agents that can benefit the mass production process.

  8. Biocontrol and plant growth-promoting activity of rhizobacteria from Chinese fields with contaminated soils.

    PubMed

    Wang, Xuefei; Mavrodi, Dmitri V; Ke, Linfeng; Mavrodi, Olga V; Yang, Mingming; Thomashow, Linda S; Zheng, Na; Weller, David M; Zhang, Jibin

    2015-05-01

    The aim of this study was to inventory the types of plant growth-promoting rhizobacteria (PGPR) present in the rhizosphere of plants grown in soils contaminated with heavy metals, recalcitrant organics, petroleum sewage or salinity in China. We screened 1223 isolates for antifungal activity and about 24% inhibited Rhizoctonia solani or Sclerotinia sclerotiorum. Twenty-four strains inhibitory to R. solani, Gaeumannomyces graminis var. tritici and/or S. sclerotiorum and representing the dominant morphotypes were assayed for PGPR activity. Seven strains contained phlD, prnD, pltC or phzF genes and produced the antibiotics 2,4-diacetylphloroglucinol, pyrrolnitrin, pyoluteorin and phenazines respectively. Six strains contained acdS, which encodes 1-aminocyclopropane-1-carboxylic acid deaminase. Phylogenetic analysis of 16S rDNA and phlD, phzF and acdS genes demonstrated that some strains identified as Pseudomonas were similar to model PGPR strains Pseudomonas protegens Pf-5, Pseudomonas chlororaphis subsp. aureofaciens 30-84 and P. brassicacearum Q8r1-96. Pseudomonas protegens- and P. chlororaphis-like strains had the greatest biocontrol activity against Rhizoctonia root rot and take-all of wheat. Pseudomonas protegens and P. brassicacearum-like strains showed the greatest promotion of canola growth. Our results indicate that strains from contaminated soils are similar to well-described PGPR found in agricultural soils worldwide.

  9. Biocontrol and plant growth-promoting activity of rhizobacteria from Chinese fields with contaminated soils

    PubMed Central

    Wang, Xuefei; Mavrodi, Dmitri V; Ke, Linfeng; Mavrodi, Olga V; Yang, Mingming; Thomashow, Linda S; Zheng, Na; Weller, David M; Zhang, Jibin

    2015-01-01

    The aim of this study was to inventory the types of plant growth-promoting rhizobacteria (PGPR) present in the rhizosphere of plants grown in soils contaminated with heavy metals, recalcitrant organics, petroleum sewage or salinity in China. We screened 1223 isolates for antifungal activity and about 24% inhibited Rhizoctonia solani or Sclerotinia sclerotiorum. Twenty-four strains inhibitory to R. solani, Gaeumannomyces graminis var. tritici and/or S. sclerotiorum and representing the dominant morphotypes were assayed for PGPR activity. Seven strains contained phlD, prnD, pltC or phzF genes and produced the antibiotics 2,4-diacetylphloroglucinol, pyrrolnitrin, pyoluteorin and phenazines respectively. Six strains contained acdS, which encodes 1-aminocyclopropane-1-carboxylic acid deaminase. Phylogenetic analysis of 16S rDNA and phlD, phzF and acdS genes demonstrated that some strains identified as Pseudomonas were similar to model PGPR strains Pseudomonas protegens Pf-5, Pseudomonas chlororaphis subsp. aureofaciens 30–84 and P. brassicacearum Q8r1-96. Pseudomonas protegens- and P. chlororaphis-like strains had the greatest biocontrol activity against Rhizoctonia root rot and take-all of wheat. Pseudomonas protegens and P. brassicacearum-like strains showed the greatest promotion of canola growth. Our results indicate that strains from contaminated soils are similar to well-described PGPR found in agricultural soils worldwide. Growth-promoting rhizobacteria in polluted soils PMID:25219642

  10. Quorum sensing is a key regulator for the antifungal and biocontrol activity of chitinase-producing Chromobacterium sp. C61.

    PubMed

    Kim, In Seon; Yang, Si Young; Park, Seur Kee; Kim, Young Cheol

    2017-01-01

    Chromobacterium sp. strain C61 has strong biocontrol activity; however, the genetic and biochemical determinants of its plant disease suppression activity are not well understood. Here, we report the identification and characterization of two new determinants of its biocontrol activity. Transposon mutagenesis was used to identify mutants that were deficient in fungal suppression. One of these mutants had an insertion in a homologue of depD, a structural gene in the dep operon, that encodes a protein involved in non-ribosomal peptide synthesis. In the second mutant, the insertion was in a homologue of the luxI gene, which encodes a homoserine lactone synthase. The luxI(-) and depD(-) mutants had no antifungal activity in vitro and a dramatically reduced capacity to suppress various plant diseases in planta. Antifungal production and biocontrol were restored by complementation of the luxI(-) mutant. Other phenotypes associated with effective biological control, including motility and lytic enzyme secretion, were also affected by the luxI mutation. Biochemical analysis of ethyl acetate extracts of culture filtrates of the mutant and wild-type strains showed that a key antifungal compound, chromobactomycin, was produced by wild-type C61 and the complemented luxI(-) mutant, but not by the luxI(-) or depD(-) mutant. These data suggest that multiple biocontrol-related phenotypes are regulated by homoserine lactones in C61. Thus, quorum sensing plays an essential role in the biological control potential of diverse bacterial lineages. © 2016 BSPP and John Wiley & Sons Ltd.

  11. Biocontrol of Bacillus subtilis against infection of Arabidopsis roots by Pseudomonas syringae is facilitated by biofilm formation and surfactin production.

    PubMed

    Bais, Harsh Pal; Fall, Ray; Vivanco, Jorge M

    2004-01-01

    Relatively little is known about the exact mechanisms used by Bacillus subtilis in its behavior as a biocontrol agent on plants. Here, we report the development of a sensitive plant infection model demonstrating that the bacterial pathogen Pseudomonas syringae pv tomato DC3000 is capable of infecting Arabidopsis roots both in vitro and in soil. Using this infection model, we demonstrated the biocontrol ability of a wild-type B. subtilis strain 6051 against P. syringae. Arabidopsis root surfaces treated with B. subtilis were analyzed with confocal scanning laser microscopy to reveal a three-dimensional B. subtilis biofilm. It is known that formation of biofilms by B. subtilis is a complex process that includes secretion of surfactin, a lipopeptide antimicrobial agent. To determine the role of surfactin in biocontrol by B. subtilis, we tested a mutant strain, M1, with a deletion in a surfactin synthase gene and, thus, deficient in surfactin production. B. subtilis M1 was ineffective as a biocontrol agent against P. syringae infectivity in Arabidopsis and also failed to form robust biofilms on either roots or inert surfaces. The antibacterial activity of surfactin against P. syringae was determined in both broth and agar cultures and also by live-dead staining methods. Although the minimum inhibitory concentrations determined were relatively high (25 microg mL(-1)), the levels of the lipopeptide in roots colonized by B. subtilis are likely to be sufficient to kill P. syringae. Our results collectively indicate that upon root colonization, B. subtilis 6051 forms a stable, extensive biofilm and secretes surfactin, which act together to protect plants against attack by pathogenic bacteria.

  12. CHARACTERIZATION AND BIOCONTROL POTENT OF STREPTOMYCES SP. ISOLATED FROM THE RHIZOSPHERE OF ONONIS ANGUSTISSIMA LAM.

    PubMed

    Ghadbane, M; Belhadj, H; Medjekal, S; Harzallah, D

    2015-01-01

    A total of 40 actinomycetes isolated from rhizosphere soils of Ononis angustissima Lam. were in vitro tested for their antagonism against deferent pathogenic microorganisms by streak assay. Among the isolates, four (21, 2A26, 1B10 and 2C34) present a potent antagonism against both pathogenic bacteria and fungi, they were selected, identified by 16S rDNA sequence analysis and phenotypic properties, and tested for their antimicrobial activity as well as their biocontrol potential against Chickpea (Cicer arietinum L.) pathogenic fungus (Fusarium oxysporum). Cultural characteristic studies strongly suggested that these strains belong to the genus Streptomyces. The four Streptomyces sp., solubilize phosphate and produce extracellular fungal cell-wall degrading enzymes chitinase and protease, as well as a marked production of acid-β-indole acetic (AIA). The nucleotide sequence of the 16S rRNA gene of Streptomyces sp. strains 21, 2A26, 1B10 and 2C34 exhibited close similarity (62-75%) with Streptomyces parvulus MARS 16S rRNA genes. The inhibition was higher against fungi and Gram+ bacteria, while Gram- bacteria were less inhibited. The growth of the plant pathogenic fungus Fusarium oxysporum was considerably inhibited in the presence of the strains 21, 2A26, 1B10 and 2C34 culture supernatant. These studies revealed that the presence of the Streptomyces strains in the soil significantly promoted the growth of the Chickpea plants. These results indicate that the Streptomyces strains isolated for rhizosphere from Ononis angustissima Lam. growing in arid conditions in southern Algeria (Sahara) could be an interesting source for antimicrobial bioactive substances and as biocontrol agents.

  13. Numerical studies of biocontrol efficacies of foliar plant pathogens in relation to the characteristics of a biocontrol agent.

    PubMed

    Xu, X-M; Salama, N; Jeffries, P; Jeger, M J

    2010-08-01

    A previously published generic mathematic model has been used in a numerical study to understand the dynamics of foliar pathogens in relation to mechanisms, and timing and coverage of biocontrol agent (BCA) applications. With the model parameter values used, it was demonstrated that a BCA possessing either competition or induced resistance as the main mechanism of biological control was more effective in reducing disease development than a BCA with either mycoparasitism or antibiosis as its mechanism. Application coverage, ranging from 50 to 90%, had little effect on biocontrol efficacy, particularly for a BCA with competition and induced resistance as the main mechanism of biocontrol. Conversely, delayed application of BCA had more profound effects on biocontrol efficacy for those with competition or induced resistance as their main mechanism than those with mycoparasitism and antibiosis. Biocontrol efficacy was greatest for a single BCA combining competition with mycoparasitism or antibiosis. The efficacy for a single BCA combining induced resistance with competition critically depended on application time; the efficacy was greatly reduced for delayed applications. The present study suggests that development of an effective strategy for BCA application is critically dependent upon our quantitative understanding of several key biocontrol processes and their interactions. Without reliable quantitative estimation of these processes, it is impossible to make quantitative predictions about biological control and hence to optimize BCA application strategies.

  14. Tailoring biocontrol to maximize top-down effects: on the importance of underlying site fertility.

    PubMed

    Hovick, Stephen M; Carson, Walter P

    2015-01-01

    The degree to which biocontrol agents impact invasive plants varies widely across landscapes, often for unknown reasons. Understanding this variability can help optimize invasive species management while also informing our understanding of trophic linkages. To address these issues, we tested three hypotheses with contrasting predictions regarding the likelihood of biocontrol success. (1) The biocontrol effort hypothesis: invasive populations are regulated primarily by top-down effects, predicting that increased biocontrol efforts alone (e.g., more individuals of a given biocontrol agent or more time since agent release) will enhance biocontrol success. (2) The relative fertility hypothesis: invasive populations are regulated primarily by bottom-up effects, predicting that nutrient enrichment will increase dominance by invasives and thus reduce biocontrol success, regardless of biocontrol efforts. (3) The fertility-dependent biocontrol effort hypothesis: top-down effects will only regulate invasive populations if bottom-up effects are weak. It predicts that greater biocontrol efforts will increase biocontrol success, but only in low-nutrient sites. To test these hypotheses, we surveyed 46 sites across three states with prior releases of Galerucella beetles, the most common biocontrol agents used against invasive purple loosestrife (Lythrum salicaria). We found strong support for the fertility-dependent biocontrol effort hypothesis, as biocontrol success occurred most often with greater biocontrol efforts, but only in low-fertility sites. This result held for early stage metrics of biocontrol success (higher Galerucella abundance) and ultimate biocontrol outcomes (decreased loosestrife plant size and abundance). Presence of the invasive grass Phalaris arundinacea was also inversely related to loosestrife abundance, suggesting that biocontrol-based reductions in loosestrife made secondary invasion by P. arundinacea more likely. Our data suggest that low-nutrient sites

  15. Transcriptional reprogramming underpins enhanced plant growth promotion by the biocontrol fungus Trichoderma hamatum GD12 during antagonistic interactions with Sclerotinia sclerotiorum in soil.

    PubMed

    Shaw, Sophie; Le Cocq, Kate; Paszkiewicz, Konrad; Moore, Karen; Winsbury, Rebecca; de Torres Zabala, Marta; Studholme, David J; Salmon, Deborah; Thornton, Christopher R; Grant, Murray R

    2016-12-01

    The free-living soil fungus Trichoderma hamatum strain GD12 is notable amongst Trichoderma strains in both controlling plant diseases and stimulating plant growth, a property enhanced during its antagonistic interactions with pathogens in soil. These attributes, alongside its markedly expanded genome and proteome compared with other biocontrol and plant growth-promoting Trichoderma strains, imply a rich potential for sustainable alternatives to synthetic pesticides and fertilizers for the control of plant disease and for increasing yields. The purpose of this study was to investigate the transcriptional responses of GD12 underpinning its biocontrol and plant growth promotion capabilities during antagonistic interactions with the pathogen Sclerotinia sclerotiorum in soil. Using an extensive mRNA-seq study capturing different time points during the pathogen-antagonist interaction in soil, we show that dynamic and biphasic signatures in the GD12 transcriptome underpin its biocontrol and plant (lettuce) growth-promoting activities. Functional predictions of differentially expressed genes demonstrate the enrichment of transcripts encoding proteins involved in transportation and oxidation-reduction reactions during both processes and an over-representation of siderophores. We identify a biphasic response during biocontrol characterized by a significant induction of transcripts encoding small-secreted cysteine-rich proteins, secondary metabolite-producing gene clusters and genes unique to GD12. These data support the hypothesis that Sclerotinia biocontrol is mediated by the synthesis and secretion of antifungal compounds and that GD12's unique reservoir of uncharacterized genes is actively recruited during the effective biological control of a plurivorous plant pathogen. © 2016 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.

  16. Characterization of Biocontrol Traits in Heterorhabditis floridensis: A Species with Broad Temperature Tolerance.

    PubMed

    Shapiro-Ilan, David I; Blackburn, Dana; Duncan, Larry; El-Borai, Fahiem E; Koppenhöfer, Heather; Tailliez, Patrick; Adams, Byron J

    2014-12-01

    Biological characteristics of two strains of the entomopathogenic nematode, Heterorhabditis floridensis (332 isolated in Florida and K22 isolated in Georgia) were described. The identity of the nematode's symbiotic bacteria was elucidated and found to be Photorhabdus luminescens subsp. luminescens. Beneficial traits pertinent to biocontrol (environmental tolerance and virulence) were characterized. The range of temperature tolerance in the H. floridensis strains was broad and showed a high level of heat tolerance. The H. floridensis strains caused higher mortality or infection in G. mellonella at 30°C and 35°C compared with S. riobrave (355), a strain widely known to be heat tolerant, and the H. floridensis strains were also capable of infecting at 17°C whereas S. riobrave (355) was not. However, at higher temperatures (37°C and 39°C), though H. floridensis readily infected G. mellonella, S. riobrave strains caused higher levels of mortality. Desiccation tolerance in H. floridensis was similar to Heterorhabditis indica (Hom1) and S. riobrave (355) and superior to S. feltiae (SN). H. bacteriophora (Oswego) and S. carpocapsae (All) exhibited higher desiccation tolerance than the H. floridensis strains. The virulence of H. floridensis to four insect pests (Aethina tumida, Conotrachelus nenuphar, Diaprepes abbreviatus, and Tenebrio molitor) was determined relative to seven other nematodes: H. bacteriophora (Oswego), H. indica (Hom1), S. carpocapsae (All), S. feltiae (SN), S. glaseri (4-8 and Vs strains), and S. riobrave (355). Virulence to A. tumida was similar among the H. floridensis strains and other nematodes except S. glaseri (Vs), S. feltiae, and S. riobrave failed to cause higher mortality than the control. Only H. bacteriophora, H. indica, S. feltiae, S. riobrave, and S. glaseri (4-8) caused higher mortality than the control in C. nenuphar. All nematodes were pathogenic to D. abbreviatus though S. glaseri (4-8) and S. riobrave (355) were the most virulent

  17. Isolation, identification, and biocontrol of antagonistic bacterium against Botrytis cinerea after tomato harvest.

    PubMed

    Shi, Jun-Feng; Sun, Chang-Qing

    2017-06-03

    Tomato is one of the most important vegetables in the world. Decay after harvest is a major issue in the development of tomato industry. Currently, the most effective method for controlling decay after harvest is storage of tomato at low temperature combined with usage of chemical bactericide; however, long-term usage of chemical bactericide not only causes pathogen resistance but also is harmful for human health and environment. Biocontrol method for the management of disease after tomato harvest has great practical significance. In this study, antagonistic bacterium B-6-1 strain was isolated from the surface of tomato and identified as Enterobacter cowanii based on morphological characteristics and physiological and biochemical features combined with sequence analysis of 16SrDNA and ropB gene and construction of dendrogram. Effects of different concentrations of antagonistic bacterium E. cowanii suspension on antifungal activity after tomato harvest were analyzed by mycelium growth rate method. Results revealed that antifungal activity was also enhanced with increasing concentrations of antagonistic bacterium; inhibitory rates of 1×10(5) colony-forming units (cfu)/mL antagonistic bacterial solution on Fusarium verticillioides, Alternaria tenuissima, and Botrytis cinerea were 46.31%, 67.48%, and 75.67%, respectively. By using in vivo inoculation method, it was further confirmed that antagonistic bacterium could effectively inhibit the occurrence of B. cinerae after tomato harvest, biocontrol effect of 1×10(9)cfu/mL zymotic fluid reached up to 95.24%, and antagonistic bacterium E. cowanii has biocontrol potential against B. cinerea after harvest of fruits and vegetables. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  18. Changing paradigms of biocontrol in the developing and the developed world

    USDA-ARS?s Scientific Manuscript database

    Microbially based biocontrol of plant diseases differs fundamentally from chemical control systems, but pesticide regulations for microbial and chemical pesticides remain similar. However, the nature and advantages of chemicals and microbials are almost totally different. Biocontrol production, dist...

  19. Biocontrol of tomato wilt disease by Bacillus subtilis isolates from natural environments depends on conserved genes mediating biofilm formation

    PubMed Central

    Liu, Hongxia; Kolter, Roberto; Losick, Richard; Guo, Jian-hua

    2014-01-01

    Summary Bacillus subtilis and other Bacilli have long been used as biological control agents against plant bacterial diseases but the mechanisms by which the bacteria confer protection are not well understood. Our goal in this study was to isolate strains of B. subtilis that exhibit high levels of biocontrol efficacy from natural environments and to investigate the mechanisms by which these strains confer plant protection. We screened a total of sixty isolates collected from various locations across China and obtained six strains that exhibited above 50% biocontrol efficacy on tomato plants against the plant pathogen Ralstonia solanacearum under greenhouse conditions. These wild strains were able to form robust biofilms both in defined medium and on tomato plant roots and exhibited strong antagonistic activities against various plant pathogens in plate assays. We show that plant protection by those strains depended on widely conserved genes required for biofilm formation, including regulatory genes and genes for matrix production. We provide evidence suggesting that matrix production is critical for bacterial colonization on plant root surfaces. Finally, we have established a model system for studies of B. subtilis-tomato plant interactions in protection against a plant pathogen. PMID:22934631

  20. Pseudomonas protegens CS1 from the lemon phyllosphere as a candidate for citrus canker biocontrol agent.

    PubMed

    Michavila, G; Adler, C; De Gregorio, P R; Lami, M J; Caram Di Santo, M C; Zenoff, A M; de Cristobal, R E; Vincent, P A

    2017-07-01

    Citrus canker is a worldwide-distributed disease caused by Xanthomonas citri subsp. citri. One of the most used strategies to control the disease is centred on copper-based compounds that cause environmental problems. Therefore, it is of interest to develop new strategies to manage the disease. Previously, we reported the ability of the siderophore pyochelin, produced by the opportunistic human pathogen Pseudomonas aeruginosa, to inhibit in vitro several bacterial species, including X. citri subsp. citri. The action mechanism, addressed with the model bacterium Escherichia coli, was connected to the generation of reactive oxygen species (ROS). This work aimed to find a non-pathogenic strain from the lemon phyllosphere that would produce pyochelin and therefore serve in canker biocontrol. An isolate that retained its capacity to colonise the lemon phyllosphere and inhibit X. citri subsp. citri was selected and characterised as Pseudomonas protegens CS1. From a liquid culture of this strain, the active compound was purified and identified as the pyochelin enantiomer, enantio-pyochelin. Using the producing strain and the pure compound, both in vitro and in vivo, we determined that the action mechanism of X. citri subsp. citri inhibition also involved the generation of ROS. Finally, the potential application of P. protegens CS1 was evaluated by spraying the bacterium in a model that mimics the natural X. citri subsp. citri infection. The ability of P. protegens CS1 to reduce canker formation makes this strain an interesting candidate as a biocontrol agent. © 2017 German Botanical Society and The Royal Botanical Society of the Netherlands.

  1. Interplay between wheat cultivars, biocontrol pseudomonads, and soil.

    PubMed

    Meyer, Joana Beatrice; Lutz, Matthias Peter; Frapolli, Michele; Péchy-Tarr, Maria; Rochat, Laurène; Keel, Christoph; Défago, Geneviève; Maurhofer, Monika

    2010-09-01

    There is a significant potential to improve the plant-beneficial effects of root-colonizing pseudomonads by breeding wheat genotypes with a greater capacity to sustain interactions with these bacteria. However, the interaction between pseudomonads and crop plants at the cultivar level, as well as the conditions which favor the accumulation of beneficial microorganisms in the wheat rhizosphere, is largely unknown. Therefore, we characterized the three Swiss winter wheat (Triticum aestivum) cultivars Arina, Zinal, and Cimetta for their ability to accumulate naturally occurring plant-beneficial pseudomonads in the rhizosphere. Cultivar performance was measured also by the ability to select for specific genotypes of 2,4-diacetylphloroglucinol (DAPG) producers in two different soils. Cultivar-specific differences were found; however, these were strongly influenced by the soil type. Denaturing gradient gel electrophoresis (DGGE) analysis of fragments of the DAPG biosynthetic gene phlD amplified from natural Pseudomonas rhizosphere populations revealed that phlD diversity substantially varied between the two soils and that there was a cultivar-specific accumulation of certain phlD genotypes in one soil but not in the other. Furthermore, the three cultivars were tested for their ability to benefit from Pseudomonas inoculants. Interestingly, Arina, which was best protected against Pythium ultimum infection by inoculation with Pseudomonas fluorescens biocontrol strain CHA0, was the cultivar which profited the least from the bacterial inoculant in terms of plant growth promotion in the absence of the pathogen. Knowledge gained of the interactions between wheat cultivars, beneficial pseudomonads, and soil types allows us to optimize cultivar-soil combinations for the promotion of growth through beneficial pseudomonads. Additionally, this information can be implemented by breeders into a new and unique breeding strategy for low-input and organic conditions.

  2. Biocontrol Agents Increase the Specific Rate of Patulin Production by Penicillium expansum but Decrease the Disease and Total Patulin Contamination of Apples.

    PubMed

    Zheng, Xiangfeng; Yang, Qiya; Zhang, Xiaoyun; Apaliya, Maurice T; Ianiri, Giuseppe; Zhang, Hongyin; Castoria, Raffaello

    2017-01-01

    Synthetic fungicides are commonly employed for the control of postharvest diseases of fruits. However, due to health concerns about the use of these chemicals, alternative control methods including biocontrol based on antagonistic yeasts are gaining in popularity. In this study, we investigated the effects of two biocontrol yeasts, Rhodotorula mucilaginosa strain 3617 and Rhodotorula kratochvilovae strain LS11, on blue mold and patulin (PAT) contamination caused by Penicillium expansum strains PY and FS7 in artificially inoculated Fuji apples stored at 20°C for 9 days. To correlate the development of the P. expansum strains in yeast-treated and untreated apples with PAT production, we quantified their biomass in the infected fruits using a recently published quantitative real-time polymerase chain reaction method based on specific primers for patF, a gene from P. expansum that is involved in PAT biosynthesis. Both yeasts significantly reduced the disease incidence caused by the two strains of P. expansum up to 5-7 days of incubation, and lowered their biomass and the progression of symptoms up to 9 days. Interestingly, both yeasts strains increased the rate of PAT production (expressed as ng patulin/μg fungal DNA) by the two pathogenic strains. Nevertheless, both biocontrol agents reduced the total PAT contamination, especially in the case of P. expansum strain FS7, the higher PAT producer of the two tested P. expansum strains. Comparing between the yeast strains, R. kratochvilovae LS11 was more effective than R. mucilaginosa 3617 for the control of P. expansum.

  3. Biocontrol Agents Increase the Specific Rate of Patulin Production by Penicillium expansum but Decrease the Disease and Total Patulin Contamination of Apples

    PubMed Central

    Zheng, Xiangfeng; Yang, Qiya; Zhang, Xiaoyun; Apaliya, Maurice T.; Ianiri, Giuseppe; Zhang, Hongyin; Castoria, Raffaello

    2017-01-01

    Synthetic fungicides are commonly employed for the control of postharvest diseases of fruits. However, due to health concerns about the use of these chemicals, alternative control methods including biocontrol based on antagonistic yeasts are gaining in popularity. In this study, we investigated the effects of two biocontrol yeasts, Rhodotorula mucilaginosa strain 3617 and Rhodotorula kratochvilovae strain LS11, on blue mold and patulin (PAT) contamination caused by Penicillium expansum strains PY and FS7 in artificially inoculated Fuji apples stored at 20°C for 9 days. To correlate the development of the P. expansum strains in yeast-treated and untreated apples with PAT production, we quantified their biomass in the infected fruits using a recently published quantitative real-time polymerase chain reaction method based on specific primers for patF, a gene from P. expansum that is involved in PAT biosynthesis. Both yeasts significantly reduced the disease incidence caused by the two strains of P. expansum up to 5–7 days of incubation, and lowered their biomass and the progression of symptoms up to 9 days. Interestingly, both yeasts strains increased the rate of PAT production (expressed as ng patulin/μg fungal DNA) by the two pathogenic strains. Nevertheless, both biocontrol agents reduced the total PAT contamination, especially in the case of P. expansum strain FS7, the higher PAT producer of the two tested P. expansum strains. Comparing between the yeast strains, R. kratochvilovae LS11 was more effective than R. mucilaginosa 3617 for the control of P. expansum. PMID:28713362

  4. Identification of Aspergillus flavus isolates as potential biocontrol agents of aflatoxin contamination in crops.

    PubMed

    Rosada, L J; Sant'anna, J R; Franco, C C S; Esquissato, G N M; Santos, P A S R; Yajima, J P R S; Ferreira, F D; Machinski, M; Corrêa, B; Castro-Prado, M A A

    2013-06-01

    Aspergillus flavus, a haploid organism found worldwide in a variety of crops, including maize, cottonseed, almond, pistachio, and peanut, causes substantial and recurrent worldwide economic liabilities. This filamentous fungus produces aflatoxins (AFLs) B1 and B2, which are among the most carcinogenic compounds from nature, acutely hepatotoxic and immunosuppressive. Recent efforts to reduce AFL contamination in crops have focused on the use of nonaflatoxigenic A. flavus strains as biological control agents. Such agents are applied to soil to competitively exclude native AFL strains from crops and thereby reduce AFL contamination. Because the possibility of genetic recombination in A. flavus could influence the stability of biocontrol strains with the production of novel AFL phenotypes, this article assesses the diversity of vegetative compatibility reactions in isolates of A. flavus to identify heterokaryon self-incompatible (HSI) strains among nonaflatoxigenic isolates, which would be used as biological controls of AFL contamination in crops. Nitrate nonutilizing (nit) mutants were recovered from 25 A. flavus isolates, and based on vegetative complementation between nit mutants and on the microscopic examination of the number of hyphal fusions, five nonaflatoxigenic (6, 7, 9 to 11) and two nontoxigenic (8 and 12) isolates of A. flavus were phenotypically characterized as HSI. Because the number of hyphal fusions is reduced in HSI strains, impairing both heterokaryon formation and the genetic exchanges with aflatoxigenic strains, the HSI isolates characterized here, especially isolates 8 and 12, are potential agents for reducing AFL contamination in crops.

  5. Biocontrol: Fungi as Nematode Control Agents

    PubMed Central

    Mankau, R.

    1980-01-01

    The fungal antagonists of nematodes consist of a great variety of organisms belonging to widely divergent orders and families of fungi. They include the nematode-trapping fungi, endoparasitic fungi, parasites of nematode eggs and cysts, and fungi which produce metabolites toxic to nematodes. The diversity, adaptations, and distribution of nematode-destroying fungi and taxonomic problems encountered in their study are reviewed. The importance of nemato-phagous fungi in soil biology, with special emphasis on their relationship to populations of plant-parasitic nematodes, is considered. While predacious fungi have long been investigated as possible biocontrol agents and have often exhibited spectacular results in vitro, their performance in field studies has generated little enthusiasm among nematologists. To date no species has demonstrated control of any plant pest to a degree achieved with nematicides, but recent studies have provided a much clearer concept of possibilities and problems in the applied use of fungal antagonists. The discovery of new species, which appear to control certain pests effectively under specific conditions, holds out some promise that fungi may be utilized as alternatives to chemical control after a more thorough and expanded study of their biology and ecology. PMID:19300699

  6. Vorticella sp: Prospective Mosquito Biocontrol Agent

    PubMed Central

    Patil, Chandrashekhar Devidas; Narkhede, Chandrakant Prakash; Suryawanshi, Rahul Khushal; Patil, Satish Vitthal

    2016-01-01

    Background: Considering the disadvantages of chemical insecticides, we aimed to evaluate Vorticella parasites for control of mosquito larvae of Anopheles stephensi and Aedes aegypti at different larval stages. Methods: Vorticella sp infected mosquito larvae were crushed in the 0.85% saline and homogenized well to get Vorticella in suspension. The effects of Vorticella sp infections on larval development were investigated by inoculating protozoan on different larval instars of An. stephensi and Ae. aegypti and observed under light microscope. Lethal time of the Vorticella infected larvae at different stages was calculated. Results: First and 2nd larval instars of both An. stephensi and Ae. aegypti did not show signs of infection by Vorticella sp., whereas 3rd instars of An. stephensi showed more Vorticella infection than those of Ae. aegypti. However, 4th larval instars of both mosquitoes were heavily infected with Vorticella parasite which was responsible for sluggish movements of larvae and eventually death. Moreover, parasites (Vorticella spp) were responsible for more than 90% reduction in adult emergence for both infected An. stephensi and Ae. aegypti. Conclusion: This study provides insights for mosquito larvicidal action of surface parasite Vorticella on different larval stages of An. stephensi and Ae. Aegypti. It could be suggested as a potential candidate in mosquito biocontrol programs. PMID:28032113

  7. Survival of the biocontrol agents Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116 on the spikes of barley in the field*

    PubMed Central

    Zhang, Xin; Zhang, Bing-xin; Zhang, Zhen; Shen, Wei-feng; Yang, Ching-hong; Yu, Jing-quan; Zhao, Yu-hua

    2005-01-01

    Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing plasmid pRP22-GFP was constructed for monitoring the colonization of two biocontrol agents, Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116, on the spikes of barley and their effect on suppression of FHB. Survival and colonization of the Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116 strains on spikes of barley were observed by tracking the bacterial transformants with GFP expression. Our field study revealed that plasmid pRP22-GFP was stably maintained in the bacterial strains without selective pressure. The retrieved GFP-tagged strains showed that the bacterial population fluctuation accorded with that of the rain events. Furthermore, both biocontrol strains gave significant protection against FHB on spikes of barley in fields. The greater suppression of barley FHB disease was resulted from the treatment of barley spikes with biocontrol agents before inoculation with F. graminearum. PMID:16052710

  8. Controlled release of Pantoea agglomerans E325 for biocontrol of fire blight disease of apple.

    PubMed

    Kim, In-Yong; Pusey, Paul Lawrence; Zhao, Youfu; Korban, Schuyler S; Choi, Hyungsoo; Kim, Kyekyoon Kevin

    2012-07-10

    Microencapsulation and controlled release of the biocontrol agent Pantoea agglomerans strain E325 (E325), an antagonist to the bacterial plant pathogen Erwinia amylovora that causes fire blight, a devastating disease of apple and pear, have been investigated. Uniform core-shell alginate microcapsules (AMCs), 60-300 μm in diameter, were fabricated to encapsulate E325 within the core, along with nutrients, to preserve viability and promote proliferation. Controlled release of E325 was achieved by separately adjusting alginate concentrations in the shell and core solutions, and by modifying the AMC size. Viability of E325 was monitored via fluorescent staining, revealing either lack of or minimal stress during or after encapsulation. Proliferation of E325 within AMCs, followed by their subsequent release, and colonization activities within confines of apple flowers were studied under different encapsulation conditions using rfp-labeled E325 to obtain highly promising results. This study provided a 'proof of concept' of the successful use of a microencapsulated biocontrol agent, E325, against E. amylovora, and could serve as a model for further studies on the development of effective plant disease management strategies.

  9. Identification of a novel fungus, Trichoderma asperellum GDFS1009, and comprehensive evaluation of its biocontrol efficacy.

    PubMed

    Wu, Qiong; Sun, Ruiyan; Ni, Mi; Yu, Jia; Li, Yaqian; Yu, Chuanjin; Dou, Kai; Ren, Jianhong; Chen, Jie

    2017-01-01

    Due to its efficient broad-spectrum antimicrobial activity, Trichoderma has been established as an internationally recognized biocontrol fungus. In this study, we found and identified a novel strain of Trichoderma asperellum, named GDFS1009. The mycelium of T. asperellum GDFS1009 exhibits a high growth rate, high sporulation capacity, and strong inhibitory effects against pathogens that cause cucumber fusarium wilt and corn stalk rot. T. asperellum GDFS1009 secretes chitinase, glucanase, and protease, which can degrade the cell walls of fungi and contribute to mycoparasitism. The secreted xylanases are good candidates for inducing plant resistance and enhancing plant immunity against pathogens. RNA sequencing (RNA-seq) and gas chromatography-mass spectrometry (GC-MS) showed that T. asperellum GDFS1009 produces primary metabolites that are precursors of antimicrobial compounds; it also produces a variety of antimicrobial secondary metabolites, including polyketides and alkanes. In addition, this study speculated the presence of six antimicrobial peptides via ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS/MS). Future studies should focus on these antimicrobial metabolites for facilitating widespread application in the field of agricultural bio-control.

  10. Identification of a novel fungus, Trichoderma asperellum GDFS1009, and comprehensive evaluation of its biocontrol efficacy

    PubMed Central

    Wu, Qiong; Sun, Ruiyan; Ni, Mi; Yu, Jia; Li, Yaqian; Yu, Chuanjin; Dou, Kai; Ren, Jianhong; Chen, Jie

    2017-01-01

    Due to its efficient broad-spectrum antimicrobial activity, Trichoderma has been established as an internationally recognized biocontrol fungus. In this study, we found and identified a novel strain of Trichoderma asperellum, named GDFS1009. The mycelium of T. asperellum GDFS1009 exhibits a high growth rate, high sporulation capacity, and strong inhibitory effects against pathogens that cause cucumber fusarium wilt and corn stalk rot. T. asperellum GDFS1009 secretes chitinase, glucanase, and protease, which can degrade the cell walls of fungi and contribute to mycoparasitism. The secreted xylanases are good candidates for inducing plant resistance and enhancing plant immunity against pathogens. RNA sequencing (RNA-seq) and gas chromatography-mass spectrometry (GC-MS) showed that T. asperellum GDFS1009 produces primary metabolites that are precursors of antimicrobial compounds; it also produces a variety of antimicrobial secondary metabolites, including polyketides and alkanes. In addition, this study speculated the presence of six antimicrobial peptides via ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS/MS). Future studies should focus on these antimicrobial metabolites for facilitating widespread application in the field of agricultural bio-control. PMID:28644879

  11. Bridgehead Effect in the Worldwide Invasion of the Biocontrol Harlequin Ladybird

    PubMed Central

    Lombaert, Eric; Guillemaud, Thomas; Cornuet, Jean-Marie; Malausa, Thibaut; Facon, Benoît; Estoup, Arnaud

    2010-01-01

    Recent studies of the routes of worldwide introductions of alien organisms suggest that many widespread invasions could have stemmed not from the native range, but from a particularly successful invasive population, which serves as the source of colonists for remote new territories. We call here this phenomenon the invasive bridgehead effect. Evaluating the likelihood of such a scenario is heuristically challenging. We solved this problem by using approximate Bayesian computation methods to quantitatively compare complex invasion scenarios based on the analysis of population genetics (microsatellite variation) and historical (first observation dates) data. We applied this approach to the Harlequin ladybird Harmonia axyridis (HA), a coccinellid native to Asia that was repeatedly introduced as a biocontrol agent without becoming established for decades. We show that the recent burst of worldwide invasions of HA followed a bridgehead scenario, in which an invasive population in eastern North America acted as the source of the colonists that invaded the European, South American and African continents, with some admixture with a biocontrol strain in Europe. This demonstration of a mechanism of invasion via a bridgehead has important implications both for invasion theory (i.e., a single evolutionary shift in the bridgehead population versus multiple changes in case of introduced populations becoming invasive independently) and for ongoing efforts to manage invasions by alien organisms (i.e., heightened vigilance against invasive bridgeheads). PMID:20305822

  12. Implications of European corn borer, Ostrinia nubilalis, infestation in an Aspergillus flavus-biocontrolled corn agroecosystem.

    PubMed

    Mencarelli, Mariangela; Accinelli, Cesare; Vicari, Alberto

    2013-09-01

    A novel biocontrol strategy consisting of field application of bioplastic-based granules inoculated with a non-toxigenic Aspergillus flavus L. strain has recently been shown to be effective for reducing aflatoxin contamination in corn. This study focused on other factors that may affect the feasibility of this biocontrol technique, and more specifically the role of the European corn borer (ECB), Ostrinia nubilalis H., in the dispersal and infestation of A. flavus in corn and its impact on crop yield. In spite of the high percentage of corn ears showing larval feeding damage, ECB-bored kernels accounted for only 3 and 4% in 2009 and 2010 respectively. Most of the damaged kernels were localised in the ear tip or immediately below. More precisely, the average incidence of ECB-bored kernels in the upper end of the ear was 32%. However, less than 5% of kernels from the central body of the ear, which includes the majority of kernels, were injured by ECB. Although ECB larvae showed a high tolerance to aflatoxin B1 and thus had the potential to serve as vectors of the mould, fungal infection of kernels was poorly associated with insect damage. ECB infestation resulted in grain yield losses not exceeding 2.5%. © 2012 Society of Chemical Industry.

  13. Phaeobacter inhibens as biocontrol agent against Vibrio vulnificus in oyster models.

    PubMed

    Porsby, Cisse Hedegaard; Gram, Lone

    2016-08-01

    Molluscan shellfish can cause food borne diseases and here we investigated if addition of Vibrio-antagonising bacteria could reduce Vibrio vulnificus in model oyster systems and prevent its establishment in live animals. Phaeobacter inhibens, which produces an antibacterial compound, tropodithietic acid (TDA), inhibited V. vulnificus as did pure TDA (MIC of 1-3.9 μM). P. inhibens DSM 17395 (at 10(6) cfu/ml) eradicated 10(5) cfu/ml V. vulnificus CMCP6 (a rifampicin resistant variant) from a co-culture oyster model system (oyster juice) whereas the pathogen grew to 10(7) cfu/ml when co-cultured with a TDA negative Phaeobacter mutant. P. inhibens grew well in oyster juice to 10(8) CFU/ml and sterile filtered samples from these cultures were inhibitory to Vibrio spp. P. inhibens established itself in live European flat oysters (Ostrea edulis) and remained at 10(5) cfu/g for five days. However, the presence of P. inhibens could not prevent subsequently added V. vulnificus from entering the live animals, likely because of too low levels of the biocontrol strain. Whilst the oyster model studies provided indication that P. inhibens DSM 17395 could be a good candidate as biocontrol agent against V. vulnificus further optimization is need in the actual animal rearing situation.

  14. Bridgehead effect in the worldwide invasion of the biocontrol harlequin ladybird.

    PubMed

    Lombaert, Eric; Guillemaud, Thomas; Cornuet, Jean-Marie; Malausa, Thibaut; Facon, Benoît; Estoup, Arnaud

    2010-03-17

    Recent studies of the routes of worldwide introductions of alien organisms suggest that many widespread invasions could have stemmed not from the native range, but from a particularly successful invasive population, which serves as the source of colonists for remote new territories. We call here this phenomenon the invasive bridgehead effect. Evaluating the likelihood of such a scenario is heuristically challenging. We solved this problem by using approximate Bayesian computation methods to quantitatively compare complex invasion scenarios based on the analysis of population genetics (microsatellite variation) and historical (first observation dates) data. We applied this approach to the Harlequin ladybird Harmonia axyridis (HA), a coccinellid native to Asia that was repeatedly introduced as a biocontrol agent without becoming established for decades. We show that the recent burst of worldwide invasions of HA followed a bridgehead scenario, in which an invasive population in eastern North America acted as the source of the colonists that invaded the European, South American and African continents, with some admixture with a biocontrol strain in Europe. This demonstration of a mechanism of invasion via a bridgehead has important implications both for invasion theory (i.e., a single evolutionary shift in the bridgehead population versus multiple changes in case of introduced populations becoming invasive independently) and for ongoing efforts to manage invasions by alien organisms (i.e., heightened vigilance against invasive bridgeheads).

  15. Biocontrol of Sugarcane Smut Disease by Interference of Fungal Sexual Mating and Hyphal Growth Using a Bacterial Isolate.

    PubMed

    Liu, Shiyin; Lin, Nuoqiao; Chen, Yumei; Liang, Zhibin; Liao, Lisheng; Lv, Mingfa; Chen, Yufan; Tang, Yingxin; He, Fei; Chen, Shaohua; Zhou, Jianuan; Zhang, Lianhui

    2017-01-01

    Sugarcane smut is a fungal disease caused by Sporisorium scitamineum, which can cause severe economic losses in sugarcane industry. The infection depends on the mating of bipolar sporida to form a dikaryon and develops into hyphae to penetrate the meristematic tissue of sugarcane. In this study, we set to isolate bacterial strains capable of blocking the fungal mating and evaluate their potential in control of sugarcane smut disease. A bacterial isolate ST4 from rhizosphere displayed potent inhibitory activity against the mating of S. scitamineum bipolar sporida and was selected for further study. Phylogenetic analyses and biochemical characterization showed that the isolate was most similar to Pseudomonas guariconensis. Methanol extracts from minimum and potato dextrose agar (PDA) agar medium, on which strain ST4 has grown, showed strong inhibitory activity on the sexual mating of S. scitamineum sporida, without killing the haploid cells MAT-1 or MAT-2. Further analysis showed that only glucose, but not sucrose, maltose, and fructose, could support strain ST4 to produce antagonistic chemicals. Consistent with the above findings, greenhouse trials showed that addition of 2% glucose to the bacterial inoculum significantly increased the strain ST4 biocontrol efficiency against sugarcane smut disease by 77% than the inoculum without glucose. The results from this study depict a new strategy to screen for biocontrol agents for control and prevention of the sugarcane smut disease.

  16. Effects of carbon concentration and carbon to nitrogen ratio on the growth and sporulation of several biocontrol fungi.

    PubMed

    Gao, Li; Sun, Man H; Liu, Xing Z; Che, Yong S

    2007-01-01

    Effects of carbon concentration and carbon to nitrogen (C:N) ratio on six biocontrol fungal strains are reported in this paper. All fungal strains had extensive growth on the media supplemented with 6-12 gl(-1) carbon and C:N ratios from 10:1 to 80:1, and differed in nutrient requirements for sporulation. Except for the two strains of Paecilomyces lilacinus, all selected fungi attained the highest spore yields at a C:N ratio of 160:1 when the carbon concentration was 12 gl(-1) for Metarhizium anisopliae SQZ-1-21, 6 gl(-1) for M. anisopliae RS-4-1 and Trichoderma viride TV-1, and 8 gl(-1) for Lecanicillium lecanii CA-1-G. The optimal conditions for P. lilacinus sporulation were 8 gl(-1) carbon with a C:N ratio of 10:1 for M-14 and 12 gl(-1) carbon with a C:N ratio of 20:1 for IPC-P, respectively. The results indicated that the influence of carbon concentration and C:N ratio on fungal growth and sporulation is strain dependent; therefore, consideration for the complexity of nutrient requirements is essential for improving yields of fungal biocontrol agents.

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

    PubMed

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

    2013-04-01

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

  18. Selection of a biocontrol agent based on a potential mechanism of action: degradation of nicotinic acid, a growth factor essential for Erwinia amylovora.

    PubMed

    Paternoster, Thomas; Défago, Geneviève; Duffy, Brion; Gessler, Cesare; Pertot, Ilaria

    2010-12-01

    This work describes a medium-based screening method for selecting microbial biocontrol agents against Erwinia amylovora based on the degradation of a specific growth factor. Erwinia amylovora, the causal agent of the devastating fire blight disease, requires nicotinic acid or nicotinamide as an essential growth factor. Potential biocontrol agents are either selected for antimicrobial production in plate or directly on immature pears or apple blossoms. In this work, we have attempted to streamline the selection of a new potential biocontrol agent with a lower risk of non-target effects by isolation based on the ability to degrade nicotinic acid in vitro, using therefore few plant materials. A total of 735 bacteria and 1237 yeast were isolated from apple blossoms and pre-screened for nicotinic acid-degradation. Pseudomonas rhizosphaerae strain JAN was able to degrade both nicotinic acid and nicotinamide. Mutants deficient in this ability were constructed. JAN, but not the mutants, controlled E. amylovora on pear slices. On detached apple blossoms, JAN colonized apple hypanthia and strongly suppressed E. amylovora growth. Under greenhouse conditions, JAN was more effective in controlling blossom blight than P. fluorescens A506, a commercial biocontrol agent of fire blight unable to degrade nicotinic acid and nicotinamide.

  19. Phytohormone profiles induced by trichoderma isolates correspond with their biocontrol and plant growth-promoting activity on melon plants.

    PubMed

    Martínez-Medina, Ainhoa; Del Mar Alguacil, Maria; Pascual, Jose A; Van Wees, Saskia C M

    2014-07-01

    The application of Trichoderma strains with biocontrol and plant growth-promoting capacities to plant substrates can help reduce the input of chemical pesticides and fertilizers in agriculture. Some Trichoderma isolates can directly affect plant pathogens, but they also are known to influence the phytohormonal network of their host plant, thus leading to an improvement of plant growth and stress tolerance. In this study, we tested whether alterations in the phytohormone signature induced by different Trichoderma isolates correspond with their ability for biocontrol and growth promotion. Four Trichoderma isolates were collected from agricultural soils and were identified as the species Trichoderma harzianum (two isolates), Trichoderma ghanense, and Trichoderma hamatum. Their antagonistic activity against the plant pathogen Fusarium oxysporum f. sp. melonis was tested in vitro, and their plant growth-promoting and biocontrol activity against Fusarium wilt on melon plants was examined in vivo, and compared to that of the commercial strain T. harzianum T-22. Several growth- and defense-related phytohormones were analyzed in the shoots of plants that were root-colonized by the different Trichoderma isolates. An increase in auxin and a decrease in cytokinins and abscisic acid content were induced by the isolates that promoted the plant growth. Principal component analysis (PCA) was used to evaluate the relationship between the plant phenotypic and hormonal variables. PCA pointed to a strong association of auxin induction with plant growth stimulation by Trichoderma. Furthermore, the disease-protectant ability of the Trichoderma strains against F. oxysporum infection seems to be more related to their induced alterations in the content of the hormones abscisic acid, ethylene, and the cytokinin trans-zeatin riboside than to the in vitro antagonism activity against F. oxysporum.

  20. Native Killer Yeasts as Biocontrol Agents of Postharvest Fungal Diseases in Lemons.

    PubMed

    Perez, María Florencia; Contreras, Luciana; Garnica, Nydia Mercedes; Fernández-Zenoff, María Verónica; Farías, María Eugenia; Sepulveda, Milena; Ramallo, Jacqueline; Dib, Julián Rafael

    2016-01-01

    Economic losses caused by postharvest diseases represent one of the main problems of the citrus industry worldwide. The major diseases affecting citrus are the "green mold" and "blue mold", caused by Penicillium digitatum and P. italicum, respectively. To control them, synthetic fungicides are the most commonly used method. However, often the emergence of resistant strains occurs and their use is becoming more restricted because of toxic effects and environmental pollution they generate, combined with trade barriers to international markets. The aim of this work was to isolate indigenous killer yeasts with antagonistic activity against fungal postharvest diseases in lemons, and to determine their control efficiency in in vitro and in vivo assays. Among 437 yeast isolates, 8.5% show to have a killer phenotype. According to molecular identification, based on the 26S rDNA D1/D2 domain sequences analysis, strains were identified belonging to the genera Saccharomyces, Wickerhamomyces, Kazachstania, Pichia, Candida and Clavispora. Killers were challenged with pathogenic molds and strains that caused the maximum in vitro inhibition of P. digitatum were selected for in vivo assays. Two strains of Pichia and one strain of Wickerhamomyces depicted a significant protection (p <0.05) from decay by P. digitatum in assays using wounded lemons. Thus, the native killer yeasts studied in this work showed to be an effective alternative for the biocontrol of postharvest fungal infections of lemons and could be promising agents for the development of commercial products for the biological control industry.

  1. Native Killer Yeasts as Biocontrol Agents of Postharvest Fungal Diseases in Lemons

    PubMed Central

    Garnica, Nydia Mercedes; Fernández-Zenoff, María Verónica; Farías, María Eugenia; Sepulveda, Milena; Ramallo, Jacqueline; Dib, Julián Rafael

    2016-01-01

    Economic losses caused by postharvest diseases represent one of the main problems of the citrus industry worldwide. The major diseases affecting citrus are the "green mold" and "blue mold", caused by Penicillium digitatum and P. italicum, respectively. To control them, synthetic fungicides are the most commonly used method. However, often the emergence of resistant strains occurs and their use is becoming more restricted because of toxic effects and environmental pollution they generate, combined with trade barriers to international markets. The aim of this work was to isolate indigenous killer yeasts with antagonistic activity against fungal postharvest diseases in lemons, and to determine their control efficiency in in vitro and in vivo assays. Among 437 yeast isolates, 8.5% show to have a killer phenotype. According to molecular identification, based on the 26S rDNA D1/D2 domain sequences analysis, strains were identified belonging to the genera Saccharomyces, Wickerhamomyces, Kazachstania, Pichia, Candida and Clavispora. Killers were challenged with pathogenic molds and strains that caused the maximum in vitro inhibition of P. digitatum were selected for in vivo assays. Two strains of Pichia and one strain of Wickerhamomyces depicted a significant protection (p <0.05) from decay by P. digitatum in assays using wounded lemons. Thus, the native killer yeasts studied in this work showed to be an effective alternative for the biocontrol of postharvest fungal infections of lemons and could be promising agents for the development of commercial products for the biological control industry. PMID:27792761

  2. Draft Genome Sequence of a Natural Root Isolate, Bacillus subtilis UD1022, a Potential Plant Growth-Promoting Biocontrol Agent

    PubMed Central

    Bishnoi, Usha

    2015-01-01

    Bacillus subtilis, which belongs to the phylum Firmicutes, is the most widely studied Gram-positive model organism. It is found in a wide variety of environments and is particularly abundant in soils and in the gastrointestinal tracts of ruminants and humans. Here, we present the complete genome sequence of the newly described B. subtilis strain UD1022. The UD1022 genome consists of a 4.025-Mbp chromosome, and other major findings from our analysis will provide insights into the genomic basis of it being a plant growth-promoting rhizobacterium (PGPR) with biocontrol potential. PMID:26159522

  3. Biological control of chestnut blight in Croatia: an interaction between host sweet chestnut, its pathogen Cryphonectria parasitica and the biocontrol agent Cryphonectria hypovirus 1.

    PubMed

    Krstin, Ljiljana; Katanić, Zorana; Ježić, Marin; Poljak, Igor; Nuskern, Lucija; Matković, Ivana; Idžojtić, Marilena; Ćurković-Perica, Mirna

    2017-03-01

    Chestnut blight, caused by the fungus Cryphonectria parasitica, is a severe chestnut disease that can be controlled with naturally occurring hypoviruses in many areas of Europe. The aim of this research was to measure the effect of different Cryphonectria hypovirus 1 (CHV1) strains on the growth of the fungal host and select strains that could potentially be used for human-mediated biocontrol in forests and orchards, and to investigate whether and how chestnut-fungus-virus interactions affect the development and growth of the lesion area on cut stems. Two Croatian CHV1 strains (CR23 and M56/1) were selected as potential biocontrol agents. The sequencing of CHV1/ORF-A showed that both of these virus strains belonged to the Italian subtype of CHV1. In vitro transfection of selected virus strains from hypovirulent to genetically diverse virus-free fungal isolates and subsequent inoculation of all virus/fungus combinations on stems of genetically diverse sweet chestnut trees revealed that Croatian virus strain CR23 had an equally hypovirulent effect on the host as the strong French strain CHV1-EP713, while M56/1 had a weaker effect. Furthermore, it was shown that in some cases the same hypovirus/fungus combinations induced various degrees of canker development on different chestnut genotypes. Some CHV1 strains belonging to the Italian subtype have similar hypovirulent effects on C. parasitica to those belonging to the French subtype. Furthermore, chestnut susceptibility and recovery could be influenced by the response of chestnut trees to particular hypovirulent C. parasitica isolates, and virus-fungus-chestnut interactions could have significant implications for the success of chestnut blight biocontrol. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  4. Cow dung extract: a medium for the growth of pseudomonads enhancing their efficiency as biofertilizer and biocontrol agent in rice.

    PubMed

    Srivastava, Rashmi; Aragno, Michel; Sharma, A K

    2010-09-01

    Some pseudomands are being utilized as biofertilizers and biopesticides because of their role in plant growth promotion and plant protection against root parasites, respectively. Two strains of Pseudomonas, P. jessenii LHRE62 and P. synxantha HHRE81, recovered from wheat rhizosphere, have shown their potential in field bioinoculation tests under rice-wheat and pulse-wheat rotation systems. Normally, pseudomonads are cultivated on synthetic media-like King's B and used for inoculation on seeds/soil drench with talcum or charcoal as carrier material. Cow dung is being used for different purposes from the ancient time and has a significant role in crop growth because of the content in humic compounds and fertilizing bioelements available in it. Here, cow dung extract was tested as a growth medium for strains LHRE62 and HHRE81, in comparison with growth in King's B medium. The log phase was delayed by 2 h as compared to growth in King's B medium. The bacterial growth yield, lower in plain cow dung extract as compared to King's B medium, was improved upon addition of different carbon substrates. Growth of rice var. Pant Dhan 4 in pot cultures was increased using liquid formulation of cow dung extract and bacteria as foliar spray, compared to their respective controls. Biocontrol efficacy of the bioagents was assessed by challenging rice crop with Rhizoctonia solani, a sheath blight pathogen. The growth promotion and biocontrol efficiencies were more pronounced in the case of mixed inocula of strains LHRE62 and HHRE81.

  5. Molecular evolution and phylogenetic analysis of biocontrol genes acquired from SCoT polymorphism of mycoparasitic Trichoderma koningii inhibiting phytopathogen Rhizoctonia solani Kuhn.

    PubMed

    Gajera, H P; Hirpara, Darshna G; Katakpara, Zinkal A; Patel, S V; Golakiya, B A

    2016-11-01

    The biocontrol agent Trichoderma (T. harzianum, T. viride, T. virens, T. hamantum, T. koningii, T. pseudokoningii and Trichoderma species) inhibited variably (15.32 - 88.12%) the in vitro growth of Rhizoctonia solani causing root rot in cotton. The T. koningii MTCC 796 evidenced highest (88.12%) growth inhibition of test pathogen followed by T. viride NBAII Tv23 (85.34%). Scanning electron microscopic study confirmed mycoparasitism for MTCC 796 and Tv23 which were capable of completely overgrowing on R. solani by degrading mycelia, coiling around the hyphae with hook-like structures. The antagonists T. harzianum NBAII Th1 and, T. virens NBAII Tvs12 exhibited strong antibiosis and formed 2-4 mm zone of inhibition for 70.28% and 46.62%, respectively growth inhibition of test pathogen. Mycoparasitism is a strong mode of action for biocontrol activity compared with antibiosis. The antagonists Trichoderma strains were performed for start codon targeted (SCoT) polymorphism to acquire biocontrol genes from potent antagonist. The six unique SCoT fragments amplified by genomic DNA of best mycoparasitic antagonist MTCC 796 strain are subjected to DNA sequencing resulted to confirm two functional sequences for activity related to biocontrol genes. The phylogenetic and molecular evolution of functional 824 bp of SCoT-3(920) and 776 bp of SCoT-6(806) fragments signify sequence homology with biocontrol genes endochitinase (partial cds of 203 amino acids) and novel hmgR genes (partial cds of 239 amino acids), respectively and the same were annotated and deposited in NCBI GenBank database. The hmgR gene is liable to be express hmg - CoA reductase which is a key enzyme for regulation of terpene biosynthesis and mycoparasitic strains produced triterpenes during antagonism to inhibit growth of fungal pathogen as evidenced with GC-MS profile. The biocontrol genes are found in best antagonist T. koningii MTCC 796 for mycoparasitic activity to restrain the growth of test pathogen R

  6. Antifungal Activity of Isolated Bacillus amyloliquefaciens SYBC H47 for the Biocontrol of Peach Gummosis

    PubMed Central

    Zhang, Yanzhou; Wei, Zhiwen; Guan, Zhengbing; Cai, Yujie; Liao, Xiangru

    2016-01-01

    The gummosis disease is caused by Botryosphaeria dothidea (Moug. ex. Fr) Ces. et de Not., and it is one of the most important diseases of stone fruits worldwide. The use of biocontrol as an alternative approach to synthetic chemical fungicides has aroused general concern about how to control plant diseases that are caused by phytopathogens. The aim of this study is to isolate Bacillus strains from raw honeys with the capacity to inhibit B. dothidea and to explore the mechanisms by which they could be used in the biocontrol of peach gummosis. Bacillus amyloliquefaciens SYBC H47 was isolated and identified on the basis of its physiological and biochemical characteristics and its 16S rRNA and gyrB gene sequences. The cell suspension and the cell-free supernatant of its culture showed significant antifungal activity against Aspergillus niger, Mucor racemosus, Fusarium oxysporum, Penicillium citrinum, and Candida albicans by agar-diffusion assays. The primary antifungal substances were bacillomycin L, fengycin, and surfactin, which were analyzed by HPLC LC/ESI-MS/MS. Bacillomycin L showed the best inhibitory effect against conidial germination of B. dothidea, followed by fengycin and surfactin. Surfactin had limited effects on mycelial growth, contrary to those of bacillomycin L and fengycin. However, a mixture of the three lipopeptides had a synergistic effect that disrupted the structure of the conidia and mycelia. In order to reduce the production cost, the use of waste frying peanut oil and soy oil as the sole carbon source increased the lipopeptide yield levels by approximately 17% (2.42 g/L) and 110% (4.35 g/L), respectively. In a field trial, the decreases in the infected gummosis rate (IGR) and the disease severity index (DSI) through cell suspension treatments were 20% and 57.5% (in 2014), respectively, and 40% and 57.5% (in 2015), respectively, in comparison with the control. In conclusion, B. amyloliquefaciens SYBC H47 could inhibit the germination of conidia

  7. Pathology smorgasboard: Biocontrol, pathogen movement, and recent fumigation results.

    USDA-ARS?s Scientific Manuscript database

    Research on soilborne pathogens, disease control, and new forest diseases of interest were presented at the Western Forest and Conservation Nursery Association meeting in 2016. Research topics included reduced-rate soil fumigation, Pythium diversity and biocontrol, pathogen movement among nurseries,...

  8. Challenges and opportunities for the commercialization of postharvest biocontrol

    USDA-ARS?s Scientific Manuscript database

    The past twenty years has seen the field of postharvest biocontrol evolve into a sophisticated science with global research programs worldwide, numerous yearly publications, patented technologies, and the development of new commercial products. The use of these products, however, still remains limi...

  9. Screening mycotoxins for quorum inhibition in a biocontrol bacterial endophyte

    USDA-ARS?s Scientific Manuscript database

    Bacterial endophytes are used as biocontrol organisms for plant pathogens such as the maize endophyte Fusarium verticillioides and its production of fumonisin mycotoxins. However, such applications are not always predictable and efficient. Bacteria communicate via cell-dependent signals, which are r...

  10. Metacridamides A and B from the biocontrol fungus metarhizium acridum

    USDA-ARS?s Scientific Manuscript database

    Metarhizium acridum, an entomopathogenic fungus, has been commercialized and used successfully for biocontrol of grasshopper pests in Africa and Australia. As part of an effort to catalog the secondary metabolites of this fungus we discovered that its conidia produce two novel 17-membered macrocycl...

  11. Grouper as a natural biocontrol of invasive lionfish.

    PubMed

    Mumby, Peter J; Harborne, Alastair R; Brumbaugh, Daniel R

    2011-01-01

    Lionfish (Pterois volitans/miles) have invaded the majority of the Caribbean region within five years. As voracious predators of native fishes with a broad habitat distribution, lionfish are poised to cause an unprecedented disruption to coral reef diversity and function. Controls of lionfish densities within its native range are poorly understood, but they have been recorded in the stomachs of large-bodied Caribbean groupers. Whether grouper predation of lionfish is sufficient to act as a biocontrol of the invasive species is unknown, but pest biocontrol by predatory fishes has been reported in other ecosystems. Groupers were surveyed along a chain of Bahamian reefs, including one of the region's most successful marine reserves which supports the top one percentile of Caribbean grouper biomass. Lionfish biomass exhibited a 7-fold and non-linear reduction in relation to the biomass of grouper. While Caribbean grouper appear to be a biocontrol of invasive lionfish, the overexploitation of their populations by fishers, means that their median biomass on Caribbean reefs is an order of magnitude less than in our study. Thus, chronic overfishing will probably prevent natural biocontrol of lionfishes in the Caribbean.

  12. Grouper as a Natural Biocontrol of Invasive Lionfish

    PubMed Central

    Mumby, Peter J.; Harborne, Alastair R.; Brumbaugh, Daniel R.

    2011-01-01

    Lionfish (Pterois volitans/miles) have invaded the majority of the Caribbean region within five years. As voracious predators of native fishes with a broad habitat distribution, lionfish are poised to cause an unprecedented disruption to coral reef diversity and function. Controls of lionfish densities within its native range are poorly understood, but they have been recorded in the stomachs of large-bodied Caribbean groupers. Whether grouper predation of lionfish is sufficient to act as a biocontrol of the invasive species is unknown, but pest biocontrol by predatory fishes has been reported in other ecosystems. Groupers were surveyed along a chain of Bahamian reefs, including one of the region's most successful marine reserves which supports the top one percentile of Caribbean grouper biomass. Lionfish biomass exhibited a 7-fold and non-linear reduction in relation to the biomass of grouper. While Caribbean grouper appear to be a biocontrol of invasive lionfish, the overexploitation of their populations by fishers, means that their median biomass on Caribbean reefs is an order of magnitude less than in our study. Thus, chronic overfishing will probably prevent natural biocontrol of lionfishes in the Caribbean. PMID:21731769

  13. Weed biocontrol in the EU: from serendipity to strategy

    USDA-ARS?s Scientific Manuscript database

    Biological control of weeds is a globally-recognized approach to the management of the worst invasive plants in the world. Unfortunately, accidental introduction of agents account for most weed biocontrol in the EU, but do include a number of current or emerging successes. From the redistribution of...

  14. Desirable Traits of a Good Biocontrol Agent against Verticillium Wilt.

    PubMed

    Deketelaere, Silke; Tyvaert, Lien; França, Soraya C; Höfte, Monica

    2017-01-01

    The soil-borne fungus Verticillium causes serious vascular disease in a wide variety of annual crops and woody perennials. Verticillium wilt is notoriously difficult to control by conventional methods, so there is great potential for biocontrol to manage this disease. In this study we aimed to review the research about Verticillium biocontrol to get a better understanding of characteristics that are desirable in a biocontrol agent (BCA) against Verticillium wilt. We only considered studies in which the BCAs were tested on plants. Most biocontrol studies were focused on plants of the Solanaceae, Malvaceae, and Brassicaceae and within these families eggplant, cotton, and oilseed rape were the most studied crops. The list of bacterial BCAs with potential against Verticillium was dominated by endophytic Bacillus and Pseudomonas isolates, while non-pathogenic xylem-colonizing Verticillium and Fusarium isolates topped the fungal list. Predominant modes of action involved in biocontrol were inhibition of primary inoculum germination, plant growth promotion, competition and induced resistance. Many BCAs showed in vitro antibiosis and mycoparasitism but these traits were not correlated with activity in vivo and there is no evidence that they play a role in planta. Good BCAs were obtained from soils suppressive to Verticillium wilt, disease suppressive composts, and healthy plants in infested fields. Desirable characteristics in a BCA against Verticillium are the ability to (1) affect the survival or germination of microsclerotia, (2) colonize the xylem and/or cortex and compete with the pathogen for nutrients and/or space, (3) induce resistance responses in the plant and/or (4) promote plant growth. Potential BCAs should be screened in conditions that resemble the field situation to increase the chance of successful use in practice. Furthermore, issues such as large scale production, formulation, preservation conditions, shelf life, and application methods should be

  15. Desirable Traits of a Good Biocontrol Agent against Verticillium Wilt

    PubMed Central

    Deketelaere, Silke; Tyvaert, Lien; França, Soraya C.; Höfte, Monica

    2017-01-01

    The soil-borne fungus Verticillium causes serious vascular disease in a wide variety of annual crops and woody perennials. Verticillium wilt is notoriously difficult to control by conventional methods, so there is great potential for biocontrol to manage this disease. In this study we aimed to review the research about Verticillium biocontrol to get a better understanding of characteristics that are desirable in a biocontrol agent (BCA) against Verticillium wilt. We only considered studies in which the BCAs were tested on plants. Most biocontrol studies were focused on plants of the Solanaceae, Malvaceae, and Brassicaceae and within these families eggplant, cotton, and oilseed rape were the most studied crops. The list of bacterial BCAs with potential against Verticillium was dominated by endophytic Bacillus and Pseudomonas isolates, while non-pathogenic xylem-colonizing Verticillium and Fusarium isolates topped the fungal list. Predominant modes of action involved in biocontrol were inhibition of primary inoculum germination, plant growth promotion, competition and induced resistance. Many BCAs showed in vitro antibiosis and mycoparasitism but these traits were not correlated with activity in vivo and there is no evidence that they play a role in planta. Good BCAs were obtained from soils suppressive to Verticillium wilt, disease suppressive composts, and healthy plants in infested fields. Desirable characteristics in a BCA against Verticillium are the ability to (1) affect the survival or germination of microsclerotia, (2) colonize the xylem and/or cortex and compete with the pathogen for nutrients and/or space, (3) induce resistance responses in the plant and/or (4) promote plant growth. Potential BCAs should be screened in conditions that resemble the field situation to increase the chance of successful use in practice. Furthermore, issues such as large scale production, formulation, preservation conditions, shelf life, and application methods should be

  16. Host-Range Dynamics of Cochliobolus lunatus: From a Biocontrol Agent to a Severe Environmental Threat

    PubMed Central

    Louis, Bengyella; Waikhom, Sayanika Devi; Roy, Pranab; Bhardwaj, Pardeep Kumar; Sharma, Chandradev K.; Singh, Mohendro Wakambam; Talukdar, Narayan Chandra

    2014-01-01

    We undertook an investigation to advance understanding of the host-range dynamics and biocontrol implications of Cochliobolus lunatus in the past decade. Potato (Solanum tuberosum L) farms were routinely surveyed for brown-to-black leaf spot disease caused by C. lunatus. A biphasic gene data set was assembled and databases were mined for reported hosts of C. lunatus in the last decade. The placement of five virulent strains of C. lunatus causing foliar necrosis of potato was studied with microscopic and phylogenetic tools. Analysis of morphology showed intraspecific variations in stromatic tissues among the virulent strains causing foliar necrosis of potato. A maximum likelihood inference based on GPDH locus separated C. lunatus strains into subclusters and revealed the emergence of unclustered strains. The evolving nutritional requirement of C. lunatus in the last decade is exhibited by the invasion of vertebrates, invertebrates, dicots, and monocots. Our results contribute towards a better understanding of the host-range dynamics of C. lunatus and provide useful implications on the threat posed to the environment when C. lunatus is used as a mycoherbicide. PMID:24987680

  17. Health economic impacts and cost-effectiveness of aflatoxin reduction strategies in Africa: Case studies in biocontrol and postharvest interventions

    PubMed Central

    Wu, Felicia; Khlangwiset, Pornsri

    2010-01-01

    Advances in health economics have proven useful in evaluating the cost-effectiveness of interventions, where the benefit usually takes the form of improved health outcomes rather than market outcomes. We perform health-based cost-effectiveness analyses of two potential aflatoxin control strategies in Africa: 1) pre-harvest biocontrol, using atoxigenic strains of Aspergillus flavus to competitively exclude toxigenic strains from colonizing maize in Nigeria, and 2) postharvest interventions in a package to reduce aflatoxin accumulation in groundnuts in Guinea. We describe how health benefits gained from each intervention, in terms of fewer aflatoxin-induced hepatocellular carcinoma (HCC) cases, can be compared with costs of implementing the interventions. We find that both interventions would be extremely cost-effective if applied widely in African agriculture. That is, the monetized value of lives saved and quality of life gained by reducing aflatoxin-induced HCC far exceeds the cost of either biocontrol or the postharvest intervention package to achieve those health benefits. The estimated cost-effectiveness ratio (CER; gross domestic product multiplied by disability-adjusted life years saved per unit cost) for biocontrol in Nigerian maize ranges from 5.10 to 24.8; while the estimated CER for the postharvest intervention package in Guinean groundnuts ranges from 0.21 to 2.08. Any intervention with a CER greater than 1 is considered by the World Health Organization to be “very cost-effective,” while an intervention with a CER greater than 0.33 is considered “cost-effective.” Aside from cost-effectiveness, public health interventions must be readily accepted by the public, and must have financial and infrastructural support to be feasible in the parts of the world where they are most needed. PMID:20234965

  18. Inhibition coefficient and molecular diversity of multi stress tolerant Trichoderma as potential biocontrol agent against Sclerotium rolfsii Sacc.

    PubMed

    Hirpara, Darshna G; Gajera, H P; Hirapara, Jaydeep G; Golakiya, B A

    2017-08-31

    Trichoderma is one of the most exploited biocontrol agent for the management of plant diseases. Twenty strains of Trichoderma (six of T. harzianum, four of T. viride, three of T. virens, three of T. koningii, each one of T. hamatum, T. reesei, T. parceramosum and Trichoderma spp.) subjected to in vitro antagonism up to 12days after inoculation against Sclerotium rolfsii Sacc. causing stem rot in groundnut. A new concept was developed to determine inhibition coefficient representing pathogen biology and biocontrol related biophysical variables. Results explained differential inhibition coefficient of test pathogen by Trichoderma antagonists. The inhibition coefficient of test pathogen was examined highest (91.13%) by T. virens NBAII Tvs12 followed by T. virens MTCC 794 (89.33%) and T. koningii MTCC 796 (62.39%). Microscopic study confirmed biocontrol mechanism as mycoparasitism for Tvs12 and antibiosis for T. koningii MTCC 796. The sclerotial biogenesis of test pathogen was elevated during weak antagonism and diminished in interactions with strong antagonists. The inhibition coefficient of S. rolfsii was significantly negatively correlated with sclerotia formation and lipid peroxidation during the antagonism. Trichoderma strains were screened for fungicides (carbendazim and tebuconazole, thiram and mancozeb) and abiotic stress (drought and salt) tolerance. Results indicated that T. koningii MTCC 796 efficiently grew better than the other strains with maximum radial growth under adverse conditions. The genetic variability among the Trichoderma was determined using 34 gene specific markers which amplified 146 alleles. The SSR similarities explained substantial diversity (15 to 87%) across Trichoderma strains and pathogen S. rolfsii. Principal coordinates analysis (PCA) were comparable to the cluster analysis and first three most informative PC components explained 64.45% of the total variation. In PCA, potent antagonists appear to be distinct from other strains. Five

  19. Influence of diluent and sample processing methods on the recovery of the biocontrol agent Pantoea agglomerans CPA-2 from different fruit surfaces.

    PubMed

    Torres, R; Viñas, I; Usall, J; Remón, D; Teixidó, N

    2012-08-01

    Determining the populations of biocontrol agents applied as a postharvest treatment on fruit surfaces is fundamental to the assessment of the microorganisms' ability to colonise and persist on fruit. To obtain maximum recovery, we must develop a methodology that involves both diluent and processing methods and that does not affect the viability of the microorganisms. The effect of diluent composition was evaluated using three diluents: phosphate buffer, peptone saline and buffered peptone saline. An additional study was performed to compare three processing methods (shaking plus sonication, stomaching and shaking plus centrifugation) on the recovery efficiency of Pantoea agglomerans strain CPA-2 from apples, oranges, nectarines and peaches treated with this biocontrol agent. Overall, slight differences occurred among diluents, although the phosphate buffer maintained the most ideal pH for CPA-2 growth (between 5.2 and 6.2). Stomaching, using the phosphate buffer as diluent, was the best procedure for recovering and enumerating the biocontrol agent; this fact suggested that no lethal effects from naturally occurring antimicrobial compounds present on the fruit skins and/or produced when the tissues were disrupted affected the recovery of the CPA-2 cells, regardless of fruit type. The growth pattern of CPA-2 on fruits maintained at 20°C and under cold conditions was similar to that obtained in previous studies, which confirms the excellent adaptation of this strain to conditions commonly used for fruit storage.

  20. Isolation of Ralstonia solanacearum-infecting bacteriophages from tomato fields in Chiang Mai, Thailand, and their experimental use as biocontrol agents.

    PubMed

    Bhunchoth, A; Phironrit, N; Leksomboon, C; Chatchawankanphanich, O; Kotera, S; Narulita, E; Kawasaki, T; Fujie, M; Yamada, T

    2015-04-01

    To isolate and characterize novel bacteriophages infecting the phytopathogen, Ralstonia solanacearum, and to evaluate them as resources with potential uses in the biocontrol of bacterial wilt. Fourteen phages infecting R. solanacearum were isolated from soil samples collected in Chiang Mai, Thailand. The phages showed different host ranges when tested against 59 R. solanacearum strains isolated from Thailand and Japan. These phages were characterized as nine podoviruses and five myoviruses based on their morphology. Podovirus J2 in combination with another podovirus (φRSB2) lysed host cells very efficiently in contaminated soil. J2 treatment prevented wilting of tomato plants infected with a highly virulent R. solanacearum strain. Treatment with J2 effectively reduced the amount of the bacterial wilt pathogen in contaminated soil and prevented bacterial wilt of tomato in pot experiments. Myovirus J6 possessed jumbo phage features, giving a unique opportunity to study its utilization as a biocontrol agent. As exemplified by J2, the phages isolated in this study represent valuable resources with potential uses in biocontrol of bacterial wilt. A rare jumbo phage J6 served as a valuable subject to understand and utilize this new group of phages. © 2015 The Society for Applied Microbiology.

  1. Experimental evidence for the phenotypic impact of admixture between wild and biocontrol Asian ladybird (Harmonia axyridis) involved in the European invasion.

    PubMed

    Turgeon, J; Tayeh, A; Facon, B; Lombaert, E; De Clercq, P; Berkvens, N; Lundgren, J G; Estoup, A

    2011-05-01

    Hybridization can fuel evolutionary processes during biological invasions. The harlequin ladybird Harmonia axyridis has long been used as a biocontrol agent before the species became invasive worldwide. Previous analysis based on microsatellite data has shown that European invasive populations bear traces of admixture between an eastern North American source, which is at the origin of the worldwide invasion, and biocontrol strains used in Europe. In this study, we tested the hypothesis that this early admixture event may have fostered the European invasion by impacting on the phenotypes of wild European populations. Mean life history traits of experimental F(1) hybrids are compared with pure parental sources and wild European crosses. Our results reveal a biased impact whereby North American beetles benefitted from being admixed with European biocontrol strains. Resemblance between experimental hybrids and wild European invasive crosses further suggests a long-lasting effect of admixture that may still be at work and fostering invasiveness. © 2011 The Authors. Journal of Evolutionary Biology © 2011 European Society For Evolutionary Biology.

  2. Pea Broth Enhances the Biocontrol Efficacy of Lysobacter capsici AZ78 by Triggering Cell Motility Associated with Biogenesis of Type IV Pilus

    PubMed Central

    Tomada, Selena; Puopolo, Gerardo; Perazzolli, Michele; Musetti, Rita; Loi, Nazia; Pertot, Ilaria

    2016-01-01

    Bacterial cells can display different types of motility, due to the presence of external appendages such as flagella and type IV pili. To date, little information on the mechanisms involved in the motility of the Lysobacter species has been available. Recently, L. capsici AZ78, a biocontrol agent of phytopathogenic oomycetes, showed the ability to move on jellified pea broth. Pea broth medium improved also the biocontrol activity of L. capsici AZ78 against Plasmopara viticola under greenhouse conditions. Noteworthy, the quantity of pea residues remaining on grapevine leaves fostered cell motility in L. capsici AZ78. Based on these results, this unusual motility related to the composition of the growth medium was investigated in bacterial strains belonging to several Lysobacter species. The six L. capsici strains tested developed dendrite-like colonies when grown on jellified pea broth, while the development of dendrite-like colonies was not recorded in the media commonly used in motility assays. To determine the presence of genes responsible for biogenesis of the flagellum and type IV pili, the genome of L. capsici AZ78 was mined. Genes encoding structural components and regulatory factors of type IV pili were upregulated in L. capsici AZ78 cells grown on the above-mentioned medium, as compared with the other tested media. These results provide new insight into the motility mechanism of L. capsici members and the role of type IV pili and pea compounds on the epiphytic fitness and biocontrol features of L. capsici AZ78. PMID:27507963

  3. The type III secretion system of biocontrol Pseudomonas fluorescens KD targets the phytopathogenic Chromista Pythium ultimum and promotes cucumber protection.

    PubMed

    Rezzonico, Fabio; Binder, Christian; Défago, Geneviève; Moënne-Loccoz, Yvan

    2005-09-01

    The type III secretion system (TTSS) is used by Proteobacteria for pathogenic or symbiotic interaction with plant and animal hosts. Recently, TTSS genes thought to originate from the phytopathogen Pseudomonas syringae were evidenced in Pseudomonas fluorescens KD, which protects cucumber from the oomycete Pythium ultimum (kingdom Chromista/Stramenopila). However, it is not known whether the TTSS contributes to plant protection by the bacterium and, if so, whether it targets the plant or the phytopathogen. Inactivation of TTSS gene hrcV following the insertion of an omega cassette strongly reduced the biocontrol activity of the pseudomonad against P. ultimum on cucumber when compared with the wild type, but had no effect on its root-colonization ability. Analysis of a plasmid-based transcriptional hrpJ'-inaZ reporter fusion revealed that expression in strain KD of the operon containing hrcV was strongly stimulated in vitro and in situ by the oomycete and not by the plant. In vitro, both strain KD and its hrcV mutant reduced the activity level of the pectinase polygalacturonase (a key pathogenicity factor) from P. ultimum, but the reduction was much stronger with the wild type. Together, these results show that the target range of bacterial TTSS is not restricted to plants and animals but also can include members of Chromista/Stramenopila, and suggest that virulence genes acquired horizontally from phytopathogenic bacteria were functionally recycled in biocontrol saprophytic Pseudomonas spp., resulting in enhanced plant protection by the latter.

  4. Effect of bacterial antagonists on lettuce: active biocontrol of Rhizoctonia solani and negligible, short-term effects on nontarget microorganisms.

    PubMed

    Scherwinski, Katja; Grosch, Rita; Berg, Gabriele

    2008-04-01

    The aim of this study was to assess the biocontrol efficacy against Rhizoctonia solani of three bacterial antagonists introduced into naturally Rhizoctonia-infested lettuce fields and to analyse their impact on the indigenous plant-associated bacteria and fungi. Lettuce seedlings were inoculated with bacterial suspensions of two endophytic strains, Serratia plymuthica 3Re4-18 and Pseudomonas trivialis 3Re2-7, and with the rhizobacterium Pseudomonas fluorescens L13-6-12 7 days before and 5 days after planting in the field. Similar statistically significant biocontrol effects were observed for all applied bacterial antagonists compared with the uninoculated control. Single-strand conformation polymorphism analysis of 16S rRNA gene or ITS1 fragments revealed a highly diverse rhizosphere and a less diverse endophytic microbial community for lettuce. Representatives of several bacterial (Alpha-, Beta- and Gammaproteobacteria, Firmicutes, Bacteriodetes), fungal (Ascomycetes, Basidiomycetes) and protist (Oomycetes) groups were present inside or on lettuce plants. Surprisingly, given that lettuce is a vegetable that is eaten raw, species of genera such as Flavobacterium, Burkholderia, Staphylococcus, Cladosporium and Aspergillus, which contain potentially human pathogenic strains, were identified. Analysis of the indigenous bacterial and endophytic fungal populations revealed only negligible, short-term effects resulting from the bacterial treatments, and that they were more influenced by field site, plant growth stage and microenvironment.

  5. Effects of rhizobacteria Paenibacillus polymyxa APEC136 and >Bacillus subtilis APEC170 on biocontrol of postharvest pathogens of apple fruits.

    PubMed

    Kim, Young Soo; Balaraju, Kotnala; Jeon, Yongho

    In this study, plant growth-promoting rhizobacteria (PGPR) were evaluated as potential biocontrol agents against postharvest pathogens of apple fruits. In vitro bioassays revealed that, out of 30 isolates screened, isolates APEC136 and APEC170 had the most significant inhibitory effects against the mycelial growth of several fungal pathogens. Analysis of 16S ribosomal RNA (rRNA) sequences identified the two effective isolates as Paenibacillus polymyxa and Bacillus subtilis, respectively. The two strains showed greater growth in brain-heart infusion broth than in other growth media. Treatment of harvested apples with suspensions of either strain reduced the symptoms of anthracnose disease caused by two fungal pathogens, Colletotrichum gloeosporioides and Colletotrichum acutatum, and white rot disease caused by Botryosphaeria dothidea. Increased productions of amylase and protease by APEC136, and increased productions of chitinase, amylase, and protease by APEC170 might have been responsible for inhibiting mycelial growth. The isolates caused a greater reduction in the growth of white rot than of anthracnose. These results indicate that the isolates APEC136 and APEC170 are promising agents for the biocontrol of anthracnose and white rot diseases in apples after harvest, and suggest that these isolates may be useful in controlling these diseases under field conditions.

  6. Effects of rhizobacteria Paenibacillus polymyxa APEC136 and Bacillus subtilis APEC170 on biocontrol of postharvest pathogens of apple fruits*

    PubMed Central

    Kim, Young Soo; Balaraju, Kotnala; Jeon, Yongho

    2016-01-01

    In this study, plant growth-promoting rhizobacteria (PGPR) were evaluated as potential biocontrol agents against postharvest pathogens of apple fruits. In vitro bioassays revealed that, out of 30 isolates screened, isolates APEC136 and APEC170 had the most significant inhibitory effects against the mycelial growth of several fungal pathogens. Analysis of 16S ribosomal RNA (rRNA) sequences identified the two effective isolates as Paenibacillus polymyxa and Bacillus subtilis, respectively. The two strains showed greater growth in brain-heart infusion broth than in other growth media. Treatment of harvested apples with suspensions of either strain reduced the symptoms of anthracnose disease caused by two fungal pathogens, Colletotrichum gloeosporioides and Colletotrichum acutatum, and white rot disease caused by Botryosphaeria dothidea. Increased productions of amylase and protease by APEC136, and increased productions of chitinase, amylase, and protease by APEC170 might have been responsible for inhibiting mycelial growth. The isolates caused a greater reduction in the growth of white rot than of anthracnose. These results indicate that the isolates APEC136 and APEC170 are promising agents for the biocontrol of anthracnose and white rot diseases in apples after harvest, and suggest that these isolates may be useful in controlling these diseases under field conditions. PMID:27921398

  7. Survival and ecological fitness of Pseudomonas fluorescens genetically engineered with dual biocontrol mechanisms.

    PubMed

    Bainton, N J; Lynch, J M; Naseby, D; Way, J A

    2004-10-01

    competitiveness around the root systems of plants even with the introduction of the Phl locus. Thus, strain Pa21 possesses the qualities necessary to provide effective integrated biocontrol, through maintaining both its wt trait of competitive exclusion on the plant roots, while also expressing the genes from the F113 biocontrol strain for Phl production. Interestingly, however, an additional beneficial trait appears to emerge with the strain Pa21's lowered survival competence compared with SBW25 EeZY6KX in the rhizosphere soil. With fears of the spread of genetically modified organisms and persistence in the soil, this trait may be of some ecological and commercial benefit and becomes a candidate for further investigation and possible exploitation.

  8. Biocontrol of the internalization of Salmonella enterica and Enterohaemorrhagic Escherichia coli in mung bean sprouts with an endophytic Bacillus subtilis.

    PubMed

    Shen, Zhenyu; Mustapha, Azlin; Lin, Mengshi; Zheng, Guolu

    2017-06-05

    Internalization of Salmonella enterica and enterohaemorrhagic Escherichia coli (EHEC) in seed sprouts poses a health risk to consumers, and the conventional sanitization methods are not always effective to reduce this risk. This study initiated a biocontrol approach to limit the internalization using endophytic Bacillus subtilis strains, which were isolated from the inner tissue of mung bean seeds or lettuce stems. By using the deferred agar method, 12 strains of B. subtilis out of 94 putative Bacillus isolates displayed inhibitory activity against at least one of the pathogenic indicators, S. enterica Typhimurium ATCC 14028 and E. coli O157:H7 505B. Two B. subtilis isolates (LCA1 and M24) showed a broad inhibitory spectrum against multiple strains of S. enterica and EHEC, Staphylococcus aureus sp., Klebsiella pneumoniae ATCC 700603, and Listeria monocytogenes Scott A, while the laboratory B. subtilis strain 168 was only moderately inhibitory against L. monocytogenes. To facilitate the tracking of the three B. subtilis strains (LCA1, M24, and 168) in the mung bean sprouts, the three strains were genetically engineered to carry the chloramphenicol acetyltransferase (cat), generating the strains LCA1-cat, M24-cat, and 168-cat, respectively. Data of the study using the cat-tagged strains demonstrated that both the two vegetable-associated and the laboratory B. subtilis strains could internalize in mung bean sprouts during the sprouting, but the latter displayed about 1.2 lg CFU/g of seeds lower in internalization. Overall, the presence of the three B. subtilis strains could significantly reduce the internalization of S. enterica or EHEC cocktail in mung bean sprouts during the sprouting. Among them, LCA1 showed the greatest inhibition against the EHEC cocktails with a reduction of about 2.0lg CFU/g of seeds by the end of sprouting (day 5), while 168 had the smallest reduction at about 0.6lg CFU/g of seeds. In addition, the three strains demonstrated a similar

  9. Reaction of Diaporthe longicolla to a strain of Sarocladium kiliense

    USDA-ARS?s Scientific Manuscript database

    Phomopsis seed decay (PSD) of soybean [Glycine max (L.) Merr.] is a seedborne fungal disease caused by Diaporthe (syn. Phomopsis) longicolla that causes yield losses and reduced seed quality. Biocontrol of this pathogen by a strain of Acremonium strictum isolated from a culture of D. longicolla was ...

  10. Postharvest biocontrol ability of killer yeasts against Monilinia fructigena and Monilinia fructicola on stone fruit.

    PubMed

    Grzegorczyk, Monika; Żarowska, Barbara; Restuccia, Cristina; Cirvilleri, Gabriella

    2017-02-01

    The antagonistic effects of Debaryomyces hansenii KI2a, D. hansenii MI1a and Wickerhamomyces anomalus BS91 were tested against Monilinia fructigena and Monilinia fructicola in in vitro and in vivo trials. All yeast strains demonstrated antifungal activity at different levels depending on species, strain and pathogen. D hansenii KI2a and W. anomalus BS91 showed the highest biocontrol activity in vitro; the production of hydrolytic enzymes, killer toxins and volatile organic compounds (VOCs) were hypothesized as their main mechanisms of action against pathogens. D hansenii KI2a and W. anomalus BS91 significantly reduced brown rot incidence and severity on peach and plum fruits artificially inoculated with M. fructigena and M. fructicola, especially when applied 24 h before pathogen inoculation. On the opposite, D. hansenii MI1a exhibited weak antagonistic activity towards M. fructigena on peach and plum fruits and was ineffective against M. fructicola. The noticeable ability of W. anomalus BS91 to control brown rot could be also correlated with its high capacity to colonize the wound tissue and to increase its population density. Accordingly, the antagonistic strains of D. hansenii and W. anomalus could be proposed as active ingredients for the development of biofungicides against Monilinia species that are responsible for considerable economic losses in stone fruit crops. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Candida parapsilosis as a Potent Biocontrol Agent against Growth and Aflatoxin Production by Aspergillus Species

    PubMed Central

    Niknejad, F; Zaini, F; Faramarzi, MA; Amini, M; Kordbacheh, P; Mahmoudi, M; Safara, M

    2012-01-01

    Background: Aflatoxin contamination of food and feed stuff is a serious health problem and significant economic concerns. In the present study, the inhibitory effect of Candida parapsilosis IP1698 on mycelial growth and aflatoxin production in aflatoxigenic strains of Aspergillus species was investigated. Methods: Mycelial growth inhibitions of nine strains of aflatoxigenic and non-aflatoxigenic Aspergillus species in the presence of C. parapsilosis investigated by pour plate technique at different pH, temperature and time of incubation. Reduction of aflatoxin was evaluated in co-cultured fungi in yeast extract sucrose broth after seven days of incubation using HPLC method. The data were analyzed by SPSS 11.5. Results: The presence of the C. parapsilosis at different pH did not affect significantly the growth rate of Aspergillus isolates. On the other hand, temperature and time of incubation showed to be significantly effective when compared to controls without C. parapsilosis (P≤0.05). In aflatoxigenic strains, minimum percentage of reductions in total aflatoxin and B1, B2, G1, G2 fractions were 92.98, 92.54, 77.48, 54.54 and 72.22 and maximum percentage of reductions were 99.59, not detectable, 94.42, and not detectable in both G1 and G2, respectively. Conclusion: C. parapsilosis might employ as a good biocontrol agent against growth and aflatoxin production by aflatoxigenic Aspergillus species PMID:23308351

  12. Can Plant Microbiome Studies Lead to Effective Biocontrol of Plant Diseases?

    PubMed

    Ellis, Jeffrey G

    2017-03-01

    In this review, the wisdom and efficacy of studies seeking disease attenuating microbes and microbiomes only in healthy plant communities is questioned and an alternative view is posited, namely that success in biocontrol of crop diseases may also come from studies of microbiota, or at least individual species isolates, associated with diseased plants. In support of this view, I summarize the current extensive knowledge of the biology behind what is probably the most successful biocontrol of a plant disease, namely the biocontrol of crown gall of stone fruit using non-pathogenic Rhizobium rhizogenes K84, in which the biocontrol agent itself came from a diseased plant.

  13. Genome Sequencing and Comparative Analysis of the Biocontrol Agent Trichoderma harzianum sensu stricto TR274

    SciTech Connect

    Steindorff, Andrei S.; Noronha, Elilane F.; Ulhoa, Cirano J.; Kuo, Alan; Salamov, Asaf A.; Haridas, Sajeet; Riley, Robert W.; Druzhinina, Irina S.; Kubicek, Christian P.; Grigoriev, Igor V.

    2015-03-17

    Biological control is a complex process which requires many mechanisms and a high diversity of biochemical pathways. The species of Trichoderma harzianum are well known for their biocontrol activity against many plant pathogens. To gain new insights into the biocontrol mechanism used by T. harzianum, we sequenced the isolate TR274 genome using Illumina. The assembly was performed using AllPaths-LG with a maximum coverage of 100x. The assembly resulted in 2282 contigs with a N50 of 37033bp. The genome size generated was 40.8 Mb and the GC content was 47.7%, similar to other Trichoderma genomes. Using the JGI Annotation Pipeline we predicted 13,932 genes with a high transcriptome support. CEGMA tests suggested 100% genome completeness and 97.9% of RNA-SEQ reads were mapped to the genome. The phylogenetic comparison using orthologous proteins with all Trichoderma genomes sequenced at JGI, corroborates the Trichoderma (T. asperellum and T. atroviride), Longibrachiatum (T. reesei and T. longibrachiatum) and Pachibasium (T. harzianum and T. virens) section division described previously. The comparison between two Trichoderma harzianum species suggests a high genome similarity but some strain-specific expansions. Analyses of the secondary metabolites, CAZymes, transporters, proteases, transcription factors were performed. The Pachybasium section expanded virtually all categories analyzed compared with the other sections, specially Longibrachiatum section, that shows a clear contraction. These results suggests that these proteins families have an important role in their respective phenotypes. Future analysis will improve the understanding of this complex genus and give some insights about its lifestyle and the interactions with the environment.

  14. Physiological characteristics of the biocontrol yeast Pichia anomala J121.

    PubMed

    Fredlund, Elisabeth; Druvefors, Ulrika; Boysen, Marianne E; Lingsten, Karl-Johan; Schnürer, Johan

    2002-08-01

    The yeast Pichia anomala J121 prevents mold spoilage and enhances preservation of moist grain in malfunctioning storage systems. Development of P. anomala J121 as a biocontrol agent requires in-depth knowledge about its physiology. P. anomala J121 grew under strictly anaerobic conditions, at temperatures between 3 degrees C and 37 degrees C, at pH values between 2.0 and 12.4, and at a water activity of 0.92 (NaCl) and 0.85 (glycerol). It could assimilate a wide range of C- and N-sources and produce killer toxin. A selective medium containing starch, nitrate, acetic acid, and chloramphenicol was developed for P. anomala. P. anomala was equally sensitive as Candida albicans to common antifungal compounds. Growth ability at a range of environmental conditions contributes to the competitive ability of the biocontrol yeast P. anomala J121.

  15. Viral biocontrol: grand experiments in disease emergence and evolution.

    PubMed

    Di Giallonardo, Francesca; Holmes, Edward C

    2015-02-01

    Although viral emergence is commonly associated with cross-species transmission, the processes and determinants of viral evolution in a novel host environment are poorly understood. We address key questions in virus emergence and evolution using data generated from two unique natural experiments: the deliberate release of myxoma virus (MYXV) and rabbit hemorrhagic disease virus (RHDV) as biological control (biocontrol) agents against the European rabbit in Australia, and which have been of enormous benefit to Australia's ecosystem and agricultural industries. Notably, although virulence evolution in MYXV and RHDV followed different trajectories, a strongly parallel evolutionary process was observed in Australia and Europe. These biocontrol agents were also characterized by a lack of transmission to nontarget host species, suggesting that there are major barriers to successful emergence. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Viral biocontrol: grand experiments in disease emergence and evolution

    PubMed Central

    Di Giallonardo, Francesca; Holmes, Edward C.

    2014-01-01

    Although viral emergence is commonly associated with cross-species transmission, the processes and determinants of viral evolution in a novel host environment are poorly understood. We address key questions in virus emergence and evolution using data generated from two unique natural experiments: the deliberate release of myxoma virus (MYXV) and rabbit hemorrhagic disease virus (RHDV) as biological control (biocontrol) agents against the European rabbit in Australia, and which have been of enormous benefit to Australia’s ecosystem and agricultural industries. Notably, although virulence evolution in MYXV and RHDV followed different trajectories, a strongly parallel evolutionary process was observed in Australia and Europe. These biocontrol agents were also characterised by a lack of transmission to non-target host species, suggesting that there are major barriers to successful emergence. PMID:25455418

  17. Characterization of endophytic strains of Bacillus mojavensis and their production of surfactin isomers

    USDA-ARS?s Scientific Manuscript database

    Bacillus subtilis consists of a large collection of strains from which several cryptic species have been delineated, and most of these along with strains within the species are important biocontrol agents. Bacillus mojavensis, a species recently distinguished from this broad Bacillus subtilis grou...

  18. From biocontrol to cancer, probiotics and beyond

    PubMed Central

    Dunne, Colum

    2013-01-01

    This invited commentary covers the period 1997–2012 and has seen changes in terminology that progressed from “basic” and “applied” to “translational” research. In the context of Bioengineered, these changes map readily onto the processes of identifying microbial characteristics appropriate for specific applications, isolation of suitable cultures, strain or genome manipulation and exploitation of these or their metabolomes across a range of settings. PMID:23247300

  19. Complete genome sequence of Bacillus velezensis M75, a biocontrol agent against fungal plant pathogens, isolated from cotton waste.

    PubMed

    Kim, Sang Yoon; Lee, Sang Yeob; Weon, Hang-Yeon; Sang, Mee Kyung; Song, Jaekyeong

    2017-01-10

    Bacillus species have been widely used as biological control agents in agricultural fields due to their ability to suppress plant pathogens. Bacillus velezensis M75 was isolated from cotton waste used for mushroom cultivation in Korea, and was found to be antagonistic to fungal plant pathogens. Here, we report the complete genome sequence of the M75 strain, which has a 4,007,450-bp single circular chromosome with 3921 genes and a G+C content of 46.60%. The genome contained operons encoding various non-ribosomal peptide synthetases and polyketide synthases, which are responsible for the biosynthesis of secondary metabolites. Our results will provide a better understanding of the genome of B. velezensis strains for their application as biocontrol agents against fungal plant pathogens in agricultural fields.

  20. Selection rhizosphere-competent microbes for development of microbial products as biocontrol agents

    NASA Astrophysics Data System (ADS)

    Mashinistova, A. V.; Elchin, A. A.; Gorbunova, N. V.; Muratov, V. S.; Kydralieva, K. A.; Khudaibergenova, B. M.; Shabaev, V. P.; Jorobekova, Sh. J.

    2009-04-01

    Rhizosphere-borne microorganisms reintroduced to the soil-root interface can establish without inducing permanent disturbance in the microbial balance and effectively colonise the rhizosphere due to carbon sources of plant root exudates. A challenge for future development of microbial products for use in agriculture will be selection of rhizosphere-competent microbes that both protect the plant from pathogens and improve crop establishment and persistence. In this study screening, collection, identification and expression of stable and technological microbial strains living in soils and in the rhizosphere of abundant weed - couch-grass Elytrigia repens L. Nevski were conducted. A total of 98 bacteria isolated from the rhizosphere were assessed for biocontrol activity in vitro against phytopathogenic fungi including Fusarium culmorum, Fusarium heterosporum, Fusarium oxysporum, Drechslera teres, Bipolaris sorokiniana, Piricularia oryzae, Botrytis cinerea, Colletothrichum atramentarium and Cladosporium sp., Stagonospora nodorum. Biocontrol activity were performed by the following methods: radial and parallel streaks, "host - pathogen" on the cuts of wheat leaves. A culture collection comprising 64 potential biocontrol agents (BCA) against wheat and barley root diseases has been established. Of these, the most effective were 8 isolates inhibitory to at least 4 out of 5 phytopathogenic fungi tested. The remaining isolates inhibited at least 1 of 5 fungi tested. Growth stimulating activity of proposed rhizobacteria-based preparations was estimated using seedling and vegetative pot techniques. Seeds-inoculation and the tests in laboratory and field conditions were conducted for different agricultural crops - wheat and barley. Intact cells, liquid culture filtrates and crude extracts of the four beneficial bacterial strains isolated from the rhizosphere of weed were studied to stimulate plant growth. As a result, four bacterial strains selected from rhizosphere of weed

  1. A mutant of the nematophagous fungus Paecilomyces lilacinus (Thom) is a novel biocontrol agent for Sclerotinia sclerotiorum.

    PubMed

    Yang, Fan; Abdelnabby, Hazem; Xiao, Yannong

    2015-12-01

    Sclerotinia sclerotiorum causes severe stem rot and yield loss in oilseed rape (Brassica napus L.) and other crops worldwide. Extensive studies have been conducted on Paecilomyces lilacinus as a nematophagous bioagent. However, no reports stated the effect of P. lilacinus as a biocontrol agent against oilseed rape rot S. sclerotiorum. This study describes such effect in lab and field trials using the new transformant pt361 derived from the wild strain P. lilacinus 36-1. Unlike the wild-type strain, the mutant pt361 showed high antagonistic effect against S. Sclerotiorum A. Under lab conditions, the pt361 inhibited (65%) radial mycelial growth of S. sclerotiorum in dual culture test producing 5.9 mm inhibition zone IZ in front of the S. sclerotiorum colony. Moreover, the cell-free filtrate of pt361 culture showed strong inhibitory effects (60.3-100%) on mycelial growth of S. sclerotiorum. In leaf detached assay, pt361 significantly (p < 0.05) inhibited (40.4-97.9%) the extension of the leaf spots caused by S. sclerotiorum A at all tested concentrations. The genomic DNA sequences of the inserted T-DNA flanking obtained from pt361 strain was cloned, verified as a glycoside hydrolase 31 family by homologous analysis with other fungal strains, and named PGH31 (2556bp). Secondary structure prediction showed a domain (Glycoside hydrolase31). Three years field trial confirmed that the cell-free filtrates or spores suspension of pt361 achieved significant (p < 0.05) suppression of oilseed rape stem rot, promoted growth and increased yield compared to the control and exceeded, at dose 100%, the action of the fungicide procymidone(®). In conclusion, the mutant pt361 of P. lilacinus is a novel and promising biocontrol agent against oilseed rape Sclerotinia stem rot. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Endophytic colonization and biocontrol performance of Pseudomonas fluorescens PICF7 in olive (Olea europaea L.) are determined neither by pyoverdine production nor swimming motility.

    PubMed

    Maldonado-González, M Mercedes; Schilirò, Elisabetta; Prieto, Pilar; Mercado-Blanco, Jesús

    2015-09-01

    Pseudomonas fluorescens PICF7 is an indigenous inhabitant of olive (Olea europaea L.) rhizosphere, able to display endophytic lifestyle in roots, to induce a wide range of defence responses upon colonization of this organ and to exert effective biological control against Verticillium wilt of olive (VWO) (Verticillium dahliae). We aimed to evaluate the involvement of specific PICF7 phenotypes in olive root colonization and VWO biocontrol effectiveness by generating mutants impaired in swimming motility (fliI) or siderophore pyoverdine production (pvdI). Besides, the performance of mutants with diminished in vitro growth in potato dextrose agar medium (gltA) and cysteine (Cys) auxotrophy was also assessed. Results showed that olive root colonization and VWO biocontrol ability of the fliI, pvdI and gltA mutants did not significantly differ from that displayed by the parental strain PICF7. Consequently, altered in vitro growth, swimming motility and pyoverdine production contribute neither to PICF7 VWO suppressive effect nor to its colonization ability. In contrast, the Cys auxotroph mutant showed reduced olive root colonization capacity and lost full biocontrol efficacy. Moreover, confocal laser scanning microscopy revealed that all mutants tested were able to endophytically colonize root tissue to the same extent as wild-type PICF7, discarding these traits as relevant for its endophytic lifestyle.

  3. Application of terminal restriction fragment length polymorphism (T-RFLP) analysis to monitor effect of biocontrol agents on rhizosphere microbial community of hot pepper (Capsicum annuum L.).

    PubMed

    Kim, Young Tae; Cho, Myoungho; Jeong, Je Yong; Lee, Hyang Burm; Kim, Seung Bum

    2010-10-01

    Microbial communities in hot pepper (Capsicum annuum L.) cultivation fields under different cultivation methods were investigated by terminal restriction fragment length polymorphism (T-RFLP) analysis. Rhizosphere soil and leaf samples were collected from control, conventional and nature-friendly cultivation fields between May and July, 2009. Two Bacillus subtilis strains were applied to nature-friendly cultivation fields as biocontrol agents during the sampling period. Relative abundances of bacteria and plant pathogenic fungi related T-RFs were also measured to monitor the effect of biocontrol agents on potential plant pathogenic fungi. In the principal component analysis (PCA) based on T-RFLP profiles, the microbial communities from rhizosphere soil samples in July, including bacteria and fungi, showed distinct difference between nature-friendly cultivation fields and other cultivation fields. However, there was no correlation between cultivation methods and leaf microbial communities at any sampling period. Changes in the abundance of bacteria related T-RF in the rhizosphere of nature-friendly cultivation fields were observed clearly two months after application of biocontrol agent, while the abundance of plant pathogenic fungi related T-RFs significantly decreased.

  4. [Effect of agrochemicals and bio-control productions on soil nematode community dynamics].

    PubMed

    Zhang, Wanmin; Duan, Yuxi; Chen, Lijie; Liang, Chen

    2002-05-01

    Dynamics of soil nematode communities amended with agrochemicals and bio-control preparations were investigated in a soybean field. The results showed that the frequency of plant non-parasitic nematodes were obviously higher in soil amended with bio-control preparations (Doufeng 1) than with urea and herbicide, however, that of plant parasitic nematodes exhibited an inverse trend.

  5. Proactive Management of Fermentation and Formulation Interactions to Improve Biocontrol Product Performance

    USDA-ARS?s Scientific Manuscript database

    The key components of biocontrol product development—discovery, fermentation, and formulation—are interactively linked to each other and ultimately to product performance. To identify biocontrol agents suited for commercial development, our discovery programs utilize a cumulative ranking system tha...

  6. Yeast microflora of nectarines and their potential for biocontrol of brown rot

    USDA-ARS?s Scientific Manuscript database

    Resident fruit microflora has been the source of biocontrol agents for the control of postharvest decays of fruits and the active ingredient in commercialized biocontrol products. With the exception of grapes and apples, information on the resident microflora of other fruits is only fragmentary; ho...

  7. Cystofilobasidium infirmominiatum as a biocontrol agent of postharvest diseases of apple and citrus

    USDA-ARS?s Scientific Manuscript database

    Two psycrotrophic yeasts isolated from the surface of lemons have been selected as biocontrol agents of the most common postharvest diseases of apples and citrus that develops during cold storage. The biocontrol yeasts were identified as Cystofilobasidium infirmominiatum and Leucosporidium scottii....

  8. Biocontrol agent Bacillus amyloliquefaciens LJ02 induces systemic resistance against cucurbits powdery mildew

    PubMed Central

    Li, Yunlong; Gu, Yilin; Li, Juan; Xu, Mingzhu; Wei, Qing; Wang, Yuanhong

    2015-01-01

    Powdery mildew is a fungal disease found in a wide range of plants and can significantly reduce crop yields. Bacterial strain LJ02 is a biocontrol agent (BCA) isolated from a greenhouse in Tianjin, China. In combination of morphological, physiological, biochemical and phylogenetic analyses, strain LJ02 was classified as a new member of Bacillus amyloliquefaciens. Greenhouse trials showed that LJ02 fermentation broth (LJ02FB) can effectively diminish the occurrence of cucurbits powdery mildew. When treated with LJ02FB, cucumber seedlings produced significantly elevated production of superoxide dismutase, peroxidase, polyphenol oxidase and phenylalanine ammonia lyase as compared to that of the control. We further confirmed that the production of free salicylic acid (SA) and expression of one pathogenesis-related (PR) gene PR-1 in cucumber leaves were markedly elevated after treating with LJ02FB, suggesting that SA-mediated defense response was stimulated. Moreover, LJ02FB-treated cucumber leaves could secrete resistance-related substances into rhizosphere that inhibit the germination of fungi spores and the growth of pathogens. Finally, we separated bacterium and its fermented substances to test their respective effects and found that both components have SA-inducing activity and bacterium plays major roles. Altogether, we identified a BCA against powdery mildew and its mode of action by inducing systemic resistance such as SA signaling pathway. PMID:26379654

  9. Biocontrol agent Bacillus amyloliquefaciens LJ02 induces systemic resistance against cucurbits powdery mildew.

    PubMed

    Li, Yunlong; Gu, Yilin; Li, Juan; Xu, Mingzhu; Wei, Qing; Wang, Yuanhong

    2015-01-01

    Powdery mildew is a fungal disease found in a wide range of plants and can significantly reduce crop yields. Bacterial strain LJ02 is a biocontrol agent (BCA) isolated from a greenhouse in Tianjin, China. In combination of morphological, physiological, biochemical and phylogenetic analyses, strain LJ02 was classified as a new member of Bacillus amyloliquefaciens. Greenhouse trials showed that LJ02 fermentation broth (LJ02FB) can effectively diminish the occurrence of cucurbits powdery mildew. When treated with LJ02FB, cucumber seedlings produced significantly elevated production of superoxide dismutase, peroxidase, polyphenol oxidase and phenylalanine ammonia lyase as compared to that of the control. We further confirmed that the production of free salicylic acid (SA) and expression of one pathogenesis-related (PR) gene PR-1 in cucumber leaves were markedly elevated after treating with LJ02FB, suggesting that SA-mediated defense response was stimulated. Moreover, LJ02FB-treated cucumber leaves could secrete resistance-related substances into rhizosphere that inhibit the germination of fungi spores and the growth of pathogens. Finally, we separated bacterium and its fermented substances to test their respective effects and found that both components have SA-inducing activity and bacterium plays major roles. Altogether, we identified a BCA against powdery mildew and its mode of action by inducing systemic resistance such as SA signaling pathway.

  10. Evaluation of yeasts from Tibetan fermented products as agents for biocontrol of blue mold of Nashi pear fruits*

    PubMed Central

    Hu, Hao; Xu, Yang; Lu, Huang-ping; Xiao, Rui; Zheng, Xiao-dong; Yu, Ting

    2015-01-01

    A total of 20 strains of yeast isolated from Tibetan fermented products were screened for antagonism against blue mold of pear caused by Penicillium expansum. Six isolates that inhibited incidence of postharvest decay by 35% or more were selected for further screening. Among them, the most effective was Rhodotorula mucilaginosa. The results showed that washed cell suspensions of R. mucilaginosa yielded better antagonistic efficacy than unwashed cell-culture mixtures, cell-free culture filtrates, and autoclaved cell cultures. Biocontrol activity improved with increasing concentrations of incubated cells. The best concentration was 1×108 cells/ml, at which the incidence of decay was only 16.7% after 6 d of incubation. The germination of conidia of P. expansum in vitro was significantly inhibited by both washed cell-suspensions and unwashed cell-culture mixtures. Rapid colonization by yeast at different concentrations showed a relationship between yeast-cell concentration and biocontrol activity. Although the titratable acidity of pear fruits increased after treatment, R. mucilaginosa did not affect the total soluble solids or ascorbic acid content. This is the first study to report that the yeast R. mucilaginosa from Tibet Autonomous Region of China may have potential as an antagonist to control the postharvest decay of pear fruits. PMID:25845361

  11. Loss of GltB Inhibits Biofilm Formation and Biocontrol Efficiency of Bacillus subtilis Bs916 by Altering the Production of γ-Polyglutamate and Three Lipopeptides.

    PubMed

    Zhou, Huafei; Luo, Chuping; Fang, Xianwen; Xiang, Yaping; Wang, Xiaoyu; Zhang, Rongsheng; Chen, Zhiyi

    2016-01-01

    This study examined the contribution of GltB on biofilm formation and biocontrol efficiency of B. subtilis Bs916. The gltB gene was identified through a biofilm phenotype screen and a bioinformatics analysis of serious biofilm formation defects, and then a gltB single knockout mutant was constructed using homologous recombination. This mutant demonstrated severe deficits in biofilm formation and colonisation along with significantly altered production ofγ-polyglutamate (γ-PGA) and three lipopeptide antibiotics (LPs) as measured by a transcriptional analysis of both the wild type B. subtilis Bs916 and the gltB mutant. Consequently, the mutant strain retained almost no antifungal activity against Rhizoctonia solani and exhibited decreased biocontrol efficiency against rice sheath blight. Very few gltB mutant cells colonised the rice stem, and they exhibited no significant nutrient chemotaxis compared to the wild type B. subtilis Bs916. The mechanism underlying these deficits in the gltB mutant appears to be decreased significantly in production of γ-PGA and a reduction in the production of both bacillomycin L and fengycin. Biofilm restoration of gltB mutant by additionγ-PGA in the EM medium demonstrated that biofilm formation was able to restore significantly at 20 g/L. GltB regulates biofilm formation by altering the production ofγ-PGA, the LPs bacillomycin L and fengcin and influences bacterial colonisation on the rice stem, which consequently leads to poor biocontrol efficiency against rice sheath blight. This is the first report of a key regulatory protein (GltB) that is involved in biofilm regulation and its regulation mechanism and biocontrol efficiency by B. subtilis.

  12. Loss of GltB Inhibits Biofilm Formation and Biocontrol Efficiency of Bacillus subtilis Bs916 by Altering the Production of γ-Polyglutamate and Three Lipopeptides

    PubMed Central

    Luo, Chuping; Fang, Xianwen; Xiang, Yaping; Wang, Xiaoyu; Zhang, Rongsheng; Chen, Zhiyi

    2016-01-01

    Aims This study examined the contribution of GltB on biofilm formation and biocontrol efficiency of B. subtilis Bs916. Methods and Results The gltB gene was identified through a biofilm phenotype screen and a bioinformatics analysis of serious biofilm formation defects, and then a gltB single knockout mutant was constructed using homologous recombination. This mutant demonstrated severe deficits in biofilm formation and colonisation along with significantly altered production ofγ-polyglutamate (γ-PGA) and three lipopeptide antibiotics (LPs) as measured by a transcriptional analysis of both the wild type B. subtilis Bs916 and the gltB mutant. Consequently, the mutant strain retained almost no antifungal activity against Rhizoctonia solani and exhibited decreased biocontrol efficiency against rice sheath blight. Very few gltB mutant cells colonised the rice stem, and they exhibited no significant nutrient chemotaxis compared to the wild type B. subtilis Bs916. The mechanism underlying these deficits in the gltB mutant appears to be decreased significantly in production of γ-PGA and a reduction in the production of both bacillomycin L and fengycin. Biofilm restoration of gltB mutant by additionγ-PGA in the EM medium demonstrated that biofilm formation was able to restore significantly at 20 g/L. Conclusions GltB regulates biofilm formation by altering the production ofγ-PGA, the LPs bacillomycin L and fengcin and influences bacterial colonisation on the rice stem, which consequently leads to poor biocontrol efficiency against rice sheath blight. Significance and Impact of Study This is the first report of a key regulatory protein (GltB) that is involved in biofilm regulation and its regulation mechanism and biocontrol efficiency by B. subtilis. PMID:27223617

  13. The Potential Use of Wolbachia-Based Mosquito Biocontrol Strategies for Japanese Encephalitis

    PubMed Central

    Jeffries, Claire L.; Walker, Thomas

    2015-01-01

    Japanese encephalitis virus (JEV) is a zoonotic pathogen transmitted by the infectious bite of Culex mosquitoes. The virus causes the development of the disease Japanese encephalitis (JE) in a small proportion of those infected, predominantly affecting children in eastern and southern Asia. Annual JE incidence estimates range from 50,000–175,000, with 25%–30% of cases resulting in mortality. It is estimated that 3 billion people live in countries in which JEV is endemic. The virus exists in an enzootic transmission cycle, with mosquitoes transmitting JEV between birds as reservoir hosts and pigs as amplifying hosts. Zoonotic infection occurs as a result of spillover events from the main transmission cycle. The reservoir avian hosts include cattle egrets, pond herons, and other species of water birds belonging to the family Ardeidae. Irrigated rice fields provide an ideal breeding ground for mosquitoes and attract migratory birds, maintaining the transmission of JEV. Although multiple vaccines have been developed for JEV, they are expensive and require multiple doses to maintain efficacy and immunity. As humans are a “dead-end” host for the virus, vaccination of the human population is unlikely to result in eradication. Therefore, vector control of the principal mosquito vector, Culex tritaeniorhynchus, represents a more promising strategy for reducing transmission. Current vector control strategies include intermittent irrigation of rice fields and space spraying of insecticides during outbreaks. However, Cx. Tritaeniorhynchus is subject to heavy exposure to pesticides in rice fields, and as a result, insecticide resistance has developed. In recent years, significant advancements have been made in the potential use of the bacterial endosymbiont Wolbachia for mosquito biocontrol. The successful transinfection of Wolbachia strains from Drosophila flies to Aedes (Stegomyia) mosquitoes has resulted in the generation of “dengue-refractory” mosquito lines

  14. Biocontrol in an impulsive predator-prey model.

    PubMed

    Terry, Alan J

    2014-10-01

    We study a model for biological pest control (or "biocontrol") in which a pest population is controlled by a program of periodic releases of a fixed yield of predators that prey on the pest. Releases are represented as impulsive increases in the predator population. Between releases, predator-pest dynamics evolve according to a predator-prey model with some fairly general properties: the pest population grows logistically in the absence of predation; the predator functional response is either of Beddington-DeAngelis type or Holling type II; the predator per capita birth rate is bounded above by a constant multiple of the predator functional response; and the predator per capita death rate is allowed to be decreasing in the predator functional response and increasing in the predator population, though the special case in which it is constant is permitted too. We prove that, when the predator functional response is of Beddington-DeAngelis type and the predators are not sufficiently voracious, then the biocontrol program will fail to reduce the pest population below a particular economic threshold, regardless of the frequency or yield of the releases. We prove also that our model possesses a pest-eradication solution, which is both locally and globally stable provided that predators are sufficiently voracious and that releases occur sufficiently often. We establish, curiously, that the pest-eradication solution can be locally stable whilst not being globally stable, the upshot of which is that, if we delay a biocontrol response to a new pest invasion, then this can change the outcome of the response from pest eradication to pest persistence. Finally, we state a number of specific examples for our model, and, for one of these examples, we corroborate parts of our analysis by numerical simulations.

  15. Modulation of Human Immune Response by Fungal Biocontrol Agents

    PubMed Central

    Konstantinovas, Cibele; de Oliveira Mendes, Tiago A.; Vannier-Santos, Marcos A.; Lima-Santos, Jane

    2017-01-01

    Although the vast majority of biological control agents is generally regarded as safe for humans and environment, the increased exposure of agriculture workers, and consumer population to fungal substances may affect the immune system. Those compounds may be associated with both intense stimulation, resulting in IgE-mediated allergy and immune downmodulation induced by molecules such as cyclosporin A and mycotoxins. This review discusses the potential effects of biocontrol fungal components on human immune responses, possibly associated to infectious, inflammatory diseases, and defective defenses. PMID:28217107

  16. Modulation of Human Immune Response by Fungal Biocontrol Agents.

    PubMed

    Konstantinovas, Cibele; de Oliveira Mendes, Tiago A; Vannier-Santos, Marcos A; Lima-Santos, Jane

    2017-01-01

    Although the vast majority of biological control agents is generally regarded as safe for humans and environment, the increased exposure of agriculture workers, and consumer population to fungal substances may affect the immune system. Those compounds may be associated with both intense stimulation, resulting in IgE-mediated allergy and immune downmodulation induced by molecules such as cyclosporin A and mycotoxins. This review discusses the potential effects of biocontrol fungal components on human immune responses, possibly associated to infectious, inflammatory diseases, and defective defenses.

  17. Biocontrol of Escherichia coli O157

    PubMed Central

    Boyacioglu, Olcay; Sharma, Manan; Sulakvelidze, Alexander; Goktepe, Ipek

    2013-01-01

    The effect of a bacteriophage cocktail (EcoShield™) that is specific against Escherichia coli O157:H7 was evaluated against a nalidixic acid-resistant enterohemorrhagic E. coli O157:H7 RM4407 (EHEC) strain on leafy greens stored under either (1) ambient air or (2) modified atmosphere (MA; 5% O2/35% CO2/60% N2). Pieces (~2 × 2 cm2) of leafy greens (lettuce and spinach) inoculated with 4.5 log CFU/cm2 EHEC were sprayed with EcoShield™ (6.5 log PFU/cm2). Samples were stored at 4 or 10°C for up to 15 d. On spinach, the level of EHEC declined by 2.38 and 2.49 log CFU/cm2 at 4 and 10°C, respectively, 30 min after phage application (p ≤ 0.05). EcoShield™ was also effective in reducing EHEC on the surface of green leaf lettuce stored at 4°C by 2.49 and 3.28 log units in 30 min and 2 h, respectively (p ≤ 0.05). At 4°C under atmospheric air, the phage cocktail significantly (p ≤ 0.05) lowered the EHEC counts in one day by 1.19, 3.21 and 3.25 log CFU/cm2 on spinach, green leaf and romaine lettuce, respectively compared with control (no bacteriophage) treatments. When stored under MA at 4°C, phages reduced (p ≤ 0.05) EHEC populations by 2.18, 3.50 and 3.13 log CFU/cm2, on spinach, green leaf and romaine lettuce. At 10°C, EHEC reductions under atmospheric air storage were 1.99, 3.90 and 3.99 log CFU/cm2 (p ≤ 0.05), while population reductions under MA were 3.08, 3.89 and 4.34 logs on spinach, green leaf and romaine lettuce, respectively, compared with controls (p ≤ 0.05). The results of this study showed that bacteriophages were effective in reducing the levels of E. coli O157:H7 on fresh leafy produce, and that the reduction was further improved when produce was stored under the MA conditions. PMID:23819107

  18. Evaluation of rhizospheric Pseudomonas and Bacillus as biocontrol tool for Xanthomonas campestris pv campestris.

    PubMed

    Mishra, Shruti; Arora, Naveen K

    2012-02-01

    Xanthomonas campestris pv campestris (Xcc), causing black rot, is one of the most yield-limiting and destructive pathogens of cruciferous crops. The intention of this study was to evaluate the potential of rhizobacteria in black rot management. Fifty-four isolates from rhizosphere soil of Brassica campestris were screened against Xcc. Two isolates namely, KA19 and SE, with inhibition radius >11 mm were selected. The combined use of them produced an average inhibition zone of 18.1 ± 1.4 mm radius (P < 0.05). 16S rRNA gene sequencing and phylogenetic analysis identified KA19 and SE as the nearest homologs (>99.4%) of Pseudomonas aeruginosa and Bacillus thuringiensis, respectively. In greenhouse study, both isolates were effective (P < 0.05) in reducing black rot lesions compared to untreated control involving either a foliar spray or the combined seed soak and soil drench. However, the combined strains (KA19 + SE) were significantly more effective (P < 0.05) when the mode of application was combined seed and soil drench. The lipid content of seeds increased significantly with the application of these strains, especially with SE alone and in combination. After 9 weeks, the Xcc population was significantly lower in soil treated with combined strains (P < 0.05). KA19 produced extracellular siderophores, influenced by various carbon sources and identified as 4-hydroxy-2-nonyl-quinoline by NMR. In Bacillus SE, two antibacterial factors corresponding to autolysins (β-N-acetylglucosaminidase) and AHL-lactonases were established. This study would strengthen our understanding for application of different rhizobacteria with various active principles like Pseudomonas and Bacillus as ingredients of a biocontrol mixture.

  19. Biocontrol of postharvest Rhizopus decay of peaches with Pichia caribbica.

    PubMed

    Xu, Baitian; Zhang, Hongyin; Chen, Keping; Xu, Qin; Yao, Yao; Gao, Hui

    2013-08-01

    A new yeast antagonist, Pichia caribbica, isolated in our laboratory from the soil collected from unsprayed orchards, was evaluated for its biocontrol capability against Rhizopus stolonifer on peaches and the possible mechanisms involved. The decay incidence and lesion diameter of Rhizopus decay of peaches treated by P. caribbica were significantly reduced compared with the control fruits, and the higher the concentration of P. caribbica, the better the efficacy of the biocontrol. Rapid colonization of the yeast in peach wounds stored at 25 °C was observed. In peaches, the activities of peroxidase (POD), catalase (CAT), and phenylalanine ammonia-lyase (PAL) were significantly induced by P. caribbica treatment compared to those of the control fruits. All these results indicated that P. caribbica has a great potential for the development of commercial formulations to control postharvest Rhizopus decay of peaches. Its modes of action were based on competition for space and nutrients with pathogens, inducement of activities of defense-related enzymes such as POD, CAT, and PAL of peaches.

  20. Efficacy of Bacillus amyloliquefaciens as biocontrol agent to fight fungal diseases of maize under tropical climates: from lab to field assays in south Kivu.

    PubMed

    Kulimushi, Parent Zihalirwa; Basime, Géant Chuma; Nachigera, Gustave Mushagalusa; Thonart, Philippe; Ongena, Marc

    2017-06-09

    In the province of South Kivu (Democratic Republic of Congo), warm and humid climatic conditions favor the development and spreading of phytopathogens. The resulting diseases cause important losses in production both in crop and after harvest. In this study, we wanted to evaluate the potential of Bacillus amyloliquefaciens as biocontrol agent to fight some newly isolated endemic fungal pathogens infesting maize. The strain S499 has been selected based on its high in vitro antagonistic activity correlating with a huge potential to secrete fungitoxic lipopeptides upon feeding on maize root exudates. Biocontrol activity of S499 was further tested on infected plantlets in growth chamber and on plants grown under field conditions over an entire cropping period. We observed a strong protective effect of this strain evaluated at two different locations with specific agro-ecological conditions. Interestingly, disease protection was associated with higher yields and our data strongly suggest that, in addition to directly inhibit pathogens, the strain may also act as biofertilizer through the solubilization of phosphorus and/or by producing plant growth hormones in the rhizosphere. This work supports the hope of exploiting such technologically advantageous bacilli for the sake of sustainable local production of this important crop in central Africa.

  1. Biocontrol activity and patulin-removal effects of Bacillus subtilis, Rhodobacter sphaeroides and Agrobacterium tumefaciens against Penicillium expansum.

    PubMed

    Wang, Y; Yuan, Y; Liu, B; Zhang, Z; Yue, T

    2016-11-01

    This study was conducted to evaluate the biocontrol potential of Bacillus subtilis CICC 10034, Rhodobacter sphaeroides CGMCC 1.2182 and Agrobacterium tumefaciens CGMCC 1.2554 against patulin (PAT)-producer Penicillium expansum and their ability to remove PAT. Bacillus subtilis effectively inhibited P. expansum both on apples and in in vitro experiments, which reduced the rot diameter on apples by 38% compared with the control. The reduction was followed by those induced by A. tumefaciens (27·63%) and R. sphaeroides (23·67%). None of the cell-free supernatant (CFS) was able to prevent pathogen growth. Three antagonists could suppress PAT production by P. expansum on apples by 98·5, 93·7 and 94·99% after treatment with B. subtilis, R. sphaeroides and A. tumefaciens respectively. In addition, the three strains led to a 0·56-1·47 log CFU g(-1) reduction in colony number of P. expansum on apples. Survival of antagonists on apple wounds revealed their tolerance to PAT. Furthermore, both live and autoclaved cells of three strains efficiently adsorbed artificially spiked PAT from medium. The selected antagonists could be applied before harvesting to control apple infection by PAT-producing fungi and also during processing to act as PAT detoxifiers. Since little information related to the capability of R. sphaeroides and A. tumefaciens to inhibit P. expansum is currently available, the results of this study provide some new perspectives to the biocontrol field. © 2016 The Society for Applied Microbiology.

  2. Bacillus amyloliquefaciens GA1 as a source of potent antibiotics and other secondary metabolites for biocontrol of plant pathogens

    PubMed Central

    2009-01-01

    Background Phytopathogenic fungi affecting crop and post-harvested vegetables are a major threat to food production and food storage. To face these drawbacks, producers have become increasingly dependent on agrochemicals. However, intensive use of these compounds has led to the emergence of pathogen resistance and severe negative environmental impacts. There are also a number of plant diseases for which chemical solutions are ineffective or non-existent as well as an increasing demand by consumers for pesticide-free food. Thus, biological control through the use of natural antagonistic microorganisms has emerged as a promising alternative to chemical pesticides for more rational and safe crop management. Results The genome of the plant-associated B. amyloliquefaciens GA1 was sample sequenced. Several gene clusters involved in the synthesis of biocontrol agents were detected. Four gene clusters were shown to direct the synthesis of the cyclic lipopeptides surfactin, iturin A and fengycin as well as the iron-siderophore bacillibactin. Beside these non-ribosomaly synthetised peptides, three additional gene clusters directing the synthesis of the antibacterial polyketides macrolactin, bacillaene and difficidin were identified. Mass spectrometry analysis of culture supernatants led to the identification of these secondary metabolites, hence demonstrating that the corresponding biosynthetic gene clusters are functional in strain GA1. In addition, genes encoding enzymes involved in synthesis and export of the dipeptide antibiotic bacilysin were highlighted. However, only its chlorinated derivative, chlorotetaine, could be detected in culture supernatants. On the contrary, genes involved in ribosome-dependent synthesis of bacteriocin and other antibiotic peptides were not detected as compared to the reference strain B. amyloliquefaciens FZB42. Conclusion The production of all of these antibiotic compounds highlights B. amyloliquefaciens GA1 as a good candidate for the

  3. The biocontrol endophytic bacterium Pseudomonas fluorescens PICF7 induces systemic defense responses in aerial tissues upon colonization of olive roots.

    PubMed

    Gómez-Lama Cabanás, Carmen; Schilirò, Elisabetta; Valverde-Corredor, Antonio; Mercado-Blanco, Jesús

    2014-01-01

    Pseudomonas fluorescens PICF7, a native olive root endophyte and effective biocontrol agent (BCA) against Verticillium wilt of olive, is able to trigger a broad range of defense responses in root tissues of this woody plant. In order to elucidate whether strain PICF7 also induces systemic defense responses in above-ground organs, aerial tissues of olive plants grown under non-gnotobiotic conditions were collected at different time points after root bacterization with this endophytic BCA. A suppression subtractive hybridization (SSH) cDNA library, enriched in up-regulated genes, was generated. This strategy enabled the identification of 376 ESTs (99 contigs and 277 singlets), many of them related to response to different stresses. Five ESTs, involved in defense responses, were selected to carry out time-course quantitative real-time PCR (qRT-PCR) experiments aiming to: (1) validate the induction of these genes, and (2) shed light on their expression pattern along time (from 1 to 15 days). Induction of olive genes potentially coding for lipoxygenase 2, catalase, 1-aminocyclopropane-1-carboxylate oxidase, and phenylananine ammonia-lyase was thus confirmed at some time points. Computational analysis also revealed that different transcription factors were up-regulated in olive aerial tissues (i.e., JERF, bHLH, WRKY), as previously reported for roots. Results confirmed that root colonization by this endophytic bacterium does not only trigger defense responses in this organ but also mounts a wide array of systemic defense responses in distant tissues (stems, leaves). This sheds light on how olive plants respond to the "non-hostile" colonization by a bacterial endophyte and how induced defense response can contribute to the biocontrol activity of strain PICF7.

  4. The biocontrol endophytic bacterium Pseudomonas fluorescens PICF7 induces systemic defense responses in aerial tissues upon colonization of olive roots

    PubMed Central

    Gómez-Lama Cabanás, Carmen; Schilirò, Elisabetta; Valverde-Corredor, Antonio; Mercado-Blanco, Jesús

    2014-01-01

    Pseudomonas fluorescens PICF7, a native olive root endophyte and effective biocontrol agent (BCA) against Verticillium wilt of olive, is able to trigger a broad range of defense responses in root tissues of this woody plant. In order to elucidate whether strain PICF7 also induces systemic defense responses in above-ground organs, aerial tissues of olive plants grown under non-gnotobiotic conditions were collected at different time points after root bacterization with this endophytic BCA. A suppression subtractive hybridization (SSH) cDNA library, enriched in up-regulated genes, was generated. This strategy enabled the identification of 376 ESTs (99 contigs and 277 singlets), many of them related to response to different stresses. Five ESTs, involved in defense responses, were selected to carry out time-course quantitative real-time PCR (qRT-PCR) experiments aiming to: (1) validate the induction of these genes, and (2) shed light on their expression pattern along time (from 1 to 15 days). Induction of olive genes potentially coding for lipoxygenase 2, catalase, 1-aminocyclopropane-1-carboxylate oxidase, and phenylananine ammonia-lyase was thus confirmed at some time points. Computational analysis also revealed that different transcription factors were up-regulated in olive aerial tissues (i.e., JERF, bHLH, WRKY), as previously reported for roots. Results confirmed that root colonization by this endophytic bacterium does not only trigger defense responses in this organ but also mounts a wide array of systemic defense responses in distant tissues (stems, leaves). This sheds light on how olive plants respond to the “non-hostile” colonization by a bacterial endophyte and how induced defense response can contribute to the biocontrol activity of strain PICF7. PMID:25250017

  5. Biosurfactants in plant-Pseudomonas interactions and their importance to biocontrol.

    PubMed

    D'aes, Jolien; De Maeyer, Katrien; Pauwelyn, Ellen; Höfte, Monica

    2010-06-01

    Production of biosurfactants is a common feature in bacteria, and in particular in plant-associated species. These bacteria include many plant beneficial and plant pathogenic Pseudomonas spp., which produce primarily cyclic lipopeptide and rhamnolipid type biosurfactants. Pseudomonas-derived biosurfactants are involved in many important bacterial functions. By modifying surface properties, biosurfactants can influence common traits such as surface motility, biofilm formation and colonization. Biosurfactants can alter the bio-availability of exogenous compounds, such as nutrients, to promote their uptake, and of endogenous metabolites, including phenazine antibiotics, resulting in an enhanced biological activity. Antibiotic activity of biosurfactants towards microbes could play a role in intraspecific competition, self-defence and pathogenesis. In addition, bacterial surfactants can affect plants in different ways, either protecting them from disease, or acting as a toxin in a plant-pathogen interaction. Biosurfactants are involved in the biocontrol activity of an increasing number of Pseudomonas strains. Consequently, further insight into the roles and activities of surfactants produced by bacteria could provide means to optimize the use of biological control as an alternative crop protection strategy.

  6. Characterization of Novel Trichoderma asperellum Isolates to Select Effective Biocontrol Agents Against Tomato Fusarium Wilt

    PubMed Central

    El_Komy, Mahmoud H.; Saleh, Amgad A.; Eranthodi, Anas; Molan, Younes Y.

    2015-01-01

    The use of novel isolates of Trichoderma with efficient antagonistic capacity against Fusarium oxysporum f. sp. lycopersici (FOL) is a promising alternative strategy to pesticides for tomato wilt management. We evaluated the antagonistic activity of 30 isolates of T. asperellum against 4 different isolates of FOL. The production of extracellular cell wall degrading enzymes of the antagonistic isolates was also measured. The random amplified polymorphic DNA (RAPD) method was applied to assess the genetic variability among the T. asperellum isolates. All of the T. asperellum isolates significantly reduced the mycelial growth of FOL isolates but the amount of growth reduction varied significantly as well. There was a correlation between the antagonistic capacity of T. asperellum isolates towards FOL and their lytic enzyme production. Isolates showing high levels of chitinase and β-1,3-glucanase activities strongly inhibited the growth of FOL isolates. RAPD analysis showed a high level of genetic variation among T. asperellum isolates. The UPGMA dendrogram revealed that T. asperellum isolates could not be grouped by their anta- gonistic behavior or lytic enzymes production. Six isolates of T. asperellum were highly antagonistic towards FOL and potentially could be used in commercial agriculture to control tomato wilt. Our results are consistent with the conclusion that understanding the genetic variation within Trichoderma isolates and their biochemical capabilities are required for the selection of effective indigenous fungal strains for the use as biocontrol agents. PMID:25774110

  7. Isolation and identification of biocontrol agent Streptomyces rimosus M527 against Fusarium oxysporum f. sp. cucumerinum.

    PubMed

    Lu, Dandan; Ma, Zheng; Xu, Xianhao; Yu, Xiaoping

    2016-08-01

    Actinomycetes have received considerable attention as biocontrol agents against fungal plant pathogens and as plant growth promoters. In this study, a total of 320 actinomycetes were isolated from various habitats in China. Among which, 77 strains have been identified as antagonistic activities against Fusarium oxysporum f. sp. cucumerinum which usually caused fusarium wilt of cucumber. Of these, isolate actinomycete M527 not only displayed broad-spectrum antifungal activity but also showed the strongest antagonistic activity against the spore germination of F. oxysporum f. sp. cucumerinum. In pot experiments, the results indicated that isolate M527 could promote the shoot growth and prevent the development of the disease on cucumber caused by F. oxysporum f. sp. cucumerinum. The control efficacy against seedling fusarium wilt of cucumber after M527 fermentation broth root-irrigation was up to 72.1% as compared to control. Based on 16S rDNA sequence analysis, the isolate M527 was identified as Streptomyces rimosus. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. DNA polymorphisms and biocontrol of Bacillus antagonistic to citrus bacterial canker with indication of the interference of phyllosphere biofilms.

    PubMed

    Huang, Tzu-Pi; Tzeng, Dean Der-Syh; Wong, Amy C L; Chen, Chun-Han; Lu, Kuan-Min; Lee, Ya-Huei; Huang, Wen-Di; Hwang, Bing-Fang; Tzeng, Kuo-Ching

    2012-01-01

    Citrus bacterial canker caused by Xanthomonas axonopodis pv. citri is a devastating disease resulting in significant crop losses in various citrus cultivars worldwide. A biocontrol agent has not been recommended for this disease. To explore the potential of bacilli native to Taiwan to control this disease, Bacillus species with a broad spectrum of antagonistic activity against various phytopathogens were isolated from plant potting mixes, organic compost and the rhizosphere soil. Seven strains TKS1-1, OF3-16, SP4-17, HSP1, WG6-14, TLB7-7, and WP8-12 showing superior antagonistic activity were chosen for biopesticide development. The genetic identity based on 16S rDNA sequences indicated that all seven native strains were close relatives of the B. subtilis group and appeared to be discrete from the B. cereus group. DNA polymorphisms in strains WG6-14, SP4-17, TKS1-1, and WP8-12, as revealed by repetitive sequence-based PCR with the BOXA1R primers were similar to each other, but different from those of the respective Bacillus type strains. However, molecular typing of the strains using either tDNA-intergenic spacer regions or 16S-23S intergenic transcribed spacer regions was unable to differentiate the strains at the species level. Strains TKS1-1 and WG6-14 attenuated symptom development of citrus bacterial canker, which was found to be correlated with a reduction in colonization and biofilm formation by X. axonopodis pv. citri on leaf surfaces. The application of a Bacillus strain TKS1-1 endospore formulation to the leaf surfaces of citrus reduced the incidence of citrus bacterial canker and could prevent development of the disease.

  9. DNA Polymorphisms and Biocontrol of Bacillus Antagonistic to Citrus Bacterial Canker with Indication of the Interference of Phyllosphere Biofilms

    PubMed Central

    Huang, Tzu-Pi; Tzeng, Dean Der-Syh; Wong, Amy C. L.; Chen, Chun-Han; Lu, Kuan-Min; Lee, Ya-Huei; Huang, Wen-Di; Hwang, Bing-Fang; Tzeng, Kuo-Ching

    2012-01-01

    Citrus bacterial canker caused by Xanthomonas axonopodis pv. citri is a devastating disease resulting in significant crop losses in various citrus cultivars worldwide. A biocontrol agent has not been recommended for this disease. To explore the potential of bacilli native to Taiwan to control this disease, Bacillus species with a broad spectrum of antagonistic activity against various phytopathogens were isolated from plant potting mixes, organic compost and the rhizosphere soil. Seven strains TKS1-1, OF3-16, SP4-17, HSP1, WG6-14, TLB7-7, and WP8-12 showing superior antagonistic activity were chosen for biopesticide development. The genetic identity based on 16S rDNA sequences indicated that all seven native strains were close relatives of the B. subtilis group and appeared to be discrete from the B. cereus group. DNA polymorphisms in strains WG6-14, SP4-17, TKS1-1, and WP8-12, as revealed by repetitive sequence-based PCR with the BOXA1R primers were similar to each other, but different from those of the respective Bacillus type strains. However, molecular typing of the strains using either tDNA-intergenic spacer regions or 16S–23S intergenic transcribed spacer regions was unable to differentiate the strains at the species level. Strains TKS1-1 and WG6-14 attenuated symptom development of citrus bacterial canker, which was found to be correlated with a reduction in colonization and biofilm formation by X. axonopodis pv. citri on leaf surfaces. The application of a Bacillus strain TKS1-1 endospore formulation to the leaf surfaces of citrus reduced the incidence of citrus bacterial canker and could prevent development of the disease. PMID:22848728

  10. In vitro biocontrol activity of halotolerant Streptomyces aureofaciens K20: A potent antagonist against Macrophomina phaseolina (Tassi) Goid.

    PubMed

    Shrivastava, P; Kumar, R; Yandigeri, M S

    2017-01-01

    A halotolerant actinobacterial strain isolated from salinity affected soil of Eastern Indo-Gangetic plains (IGP), Uttar Pradesh, India, was characterised for its antagonistic potential against Macrophomina phaseolina by dual-culture assay. It was shown to effectively inhibit the growth of M. phaseolina with an inhibition zone of 27 ± 1.33 mm. Further the actinobacterial strain was evaluated for its plant growth promoting (PGP) properties and its ability to produce biocontrol related extracellular enzymes viz. amylase, protease, cellulase, chitinase, gelatinase and urease. The results revealed that the actinobacterial strain had PGP potential along with positive assay for amylase, chitinase and urease. The interaction study between antagonist strain and fungal pathogen, performed by scanning electron microscopy technique revealed that the actinobacterium was able to damage fungal mycelia may be due to chitinase, establishing its role as a potential antagonist against M. phaseolina. The actinobacterial isolate was characterised by 16S rDNA gene sequencing, and was identified as Streptomyces genera. The identified gene sequence was deposited to NCBI GenBank with an accession number KP331758.

  11. Health economic impacts and cost-effectiveness of aflatoxin-reduction strategies in Africa: case studies in biocontrol and post-harvest interventions.

    PubMed

    Wu, F; Khlangwiset, P

    2010-04-01

    Advances in health economics have proven useful in evaluating the cost-effectiveness of interventions, where the benefit usually takes the form of improved health outcomes rather than market outcomes. The paper performs health-based cost-effectiveness analyses of two potential aflatoxin control strategies in Africa: (1) pre-harvest biocontrol, using atoxigenic strains of Aspergillus flavus competitively to exclude toxigenic strains from colonizing maize in Nigeria; and (2) post-harvest interventions in a package to reduce aflatoxin accumulation in groundnuts in Guinea. It is described how health benefits gained from each intervention, in terms of fewer aflatoxin-induced hepatocellular carcinoma cases, can be compared with the costs of implementing the interventions. It is found that both interventions would be extremely cost-effective if applied widely in African agriculture. That is, the monetized value of lives saved and quality of life gained by reducing aflatoxin-induced hepatocellular carcinoma far exceeds the cost of either biocontrol or the post-harvest intervention package to achieve those health benefits. The estimated cost-effectiveness ratio (CER; gross domestic product multiplied by disability-adjusted life years saved per unit cost) for biocontrol in Nigerian maize ranged from 5.10 to 24.8; while the estimated CER for the post-harvest intervention package in Guinean groundnuts ranged from 0.21 to 2.08. Any intervention with a CER > 1 is considered by the World Health Organization (WHO) to be 'very cost-effective', while an intervention with a CER > 0.33 is considered 'cost-effective'. Aside from cost-effectiveness, public health interventions must be readily accepted by the public, and must have financial and infrastructural support to be feasible in the parts of the world where they are most needed.

  12. Liquid cultivation techniques for enhancing the effectiveness of yeast biocontrol agents

    USDA-ARS?s Scientific Manuscript database

    Biocontrol agents will gain broader acceptance into integrated pest management programs if products with enhanced efficacy and consistency of performance are developed. Though given comparatively little research attention, liquid cultivation environments and protocols can and should be optimized to...

  13. Proactive management of fermentation and formulation interactions to improve biocontrol product performance

    USDA-ARS?s Scientific Manuscript database

    The key components of biocontrol product development; discovery, fermentation, and formulation, are interactively linked to each other and ultimately, to product performance. The fermentation environment can be managed to maximize the quantity and quality of biomass and bioproducts produced which, ...

  14. Phyllostictines A-D, Oxazatricycloalkenones Produced by Phyllosticta cirsii, A Potential Mycoherbicide for Cirsium arvense Biocontrol

    USDA-ARS?s Scientific Manuscript database

    Phyllosticta cirsii, a fungal pathogen isolated from Cirsium arvense and proposed as biocontrol agent of this noxious perennial weed, produces in liquid cultures different phytotoxic metabolites with potential herbicidal activity. Four new oxazatricycloalkenones, named phyllostictines A-D, were isol...

  15. Risk assessment: progress of quarantine biocontrol research on Chinese Tallow, Melaleuca, and Downy Rose Myrtle

    USDA-ARS?s Scientific Manuscript database

    Risk assessments of two biocontrol candidates for Chinese tallow, Triadica sebifera (Euphoriales: Euphorbiaceae), and one for Melaleuca, Melaleuca quinquenervia (Myrtales: Myrtaceae), were conducted during 2009 and continuing into 2010 by USDA scientists located at the Florida Department of Agricul...

  16. Evaluation of the atoxigenic Aspergillus flavus strain AF36 in pistachio orchards

    USDA-ARS?s Scientific Manuscript database

    The atoxigenic strain Aspergillus flavus AF36, which has been extensively used as a biocontrol agent in commercial corn and cotton fields to reduce aflatoxin contamination, was applied in research pistachio orchards from 2002 to 2005 and in commercial pistachio orchards from 2008 to 2011. AF36 was a...

  17. Detection and quantification by PCR assay of the biocontrol agent Pantoea agglomerans CPA-2 on apples.

    PubMed

    Soto-Muñoz, Lourdes; Teixidó, Neus; Usall, Josep; Viñas, Inmaculada; Torres, Rosario

    2014-04-03

    The registration of biological control agents requires the development of monitoring systems to detect and quantify the agent in the environment. Pantoea agglomerans CPA-2 is an effective biocontrol agent for postharvest diseases of citrus and pome fruits. The monitoring of CPA-2 in postharvest semi-commercial trials was evaluated by Rodac impression plates and the colonies isolated were confirmed by conventional PCR using the SCAR primers PAGA1 and PAGB1. Samples were taken from different surfaces that had contact with CPA-2, the surrounding environment and working clothes worn by handlers. Moreover, population dynamics of the strain CPA-2 were determined on apple surfaces using both the classical plating technique and real-time quantitative PCR (qPCR). A qPCR assay using a 3'-minor groove-binding (MGB) probe was developed for the specific detection and quantification of P. agglomerans strain CPA-2. Based on the nucleotide sequence of a SCAR fragment of CPA-2, one primer set and TaqMan MGB probe were designed. The primers SP2-F/SP2-R and the TaqMan MGB probe showed a specific detection of strain CPA-2 on apple surfaces, which was verified tested against purified DNA from 17 strains of P. agglomerans, 4 related Pantoea species, and 21 bacterial strains from other genera isolated from whole and also freshly-cut fruit and vegetables. The detection level was approximately 10(3) cells per reaction, and the standard curve was linear within a range of 5log units. Results from semi-commercial trials showed that CPA-2 had a low impact. The maximum persistence of P. agglomerans CPA-2 was not longer than 5days in plastic boxes stored at 0°C. Significant differences in CPA-2 population level dynamics were observed in results obtained by qPCR and dilution plating. These differences may indicate the presence of non-degraded DNA from non-viable cells. In conclusion, qPCR is a novel potential tool to quickly and specifically monitor recent surface colonisation by CPA-2

  18. Biocontrol of fouling pests: Effect of diversity, identity and density of control agents.

    PubMed

    Atalah, Javier; Newcombe, Emma M; Zaiko, Anastasija

    2016-04-01

    Augmentative biocontrol, using native natural enemies, has been suggested as a promising tool to control marine biofouling pests on artificial structures. However, there are still important knowledge gaps to be addressed before biocontrol can be considered as a management tool. In a field experiment on floating marine structures we examined intra- and interspecific consumer interactions among biocontrol agents on different surface orientations. We tested the effect of identity, density and diversity of three invertebrates (the 11-arm seastar Coscinasterias muricata, the sea urchin Evechinus chloroticus and the gastropod Cook's turban Cookia sulcata) to reduce established biofouling and to prevent fouling growth on defouled surfaces. High densities of biocontrol agents were not more effective at fouling control (cover and biomass) than low densities. Nor did multi-species treatments function more effectively than mono-specific ones. However, biocontrol agent identity was important, with the 11-arm seastar and Cook's turban being the most effective at fouling reduction and prevention, respectively. Surface orientation had a strong effect on the effectiveness of control agents, with the best results obtained on vertical compared to diagonal and underside surfaces. This study confirmed the potential of biocontrol as a management tool for marine pest, indicating that identity is more important than richness and density of control agents. It also highlighted the limitations of this approach on diagonal and underside surfaces, where control agents have limited retention ability. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. The effect of locust bean gum (LBG)-based edible coatings carrying biocontrol yeasts against Penicillium digitatum and Penicillium italicum causal agents of postharvest decay of mandarin fruit.

    PubMed

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

    2016-09-01

    Strains belonging to Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans species were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic molds Penicillium digitatum and Penicillium italicum. Moreover, studies aimed at screening the antifungal activity of selected yeast strains in vivo conditions against P. digitatum and P. italicum, and investigated the efficacy of a polysaccharidic matrix, locust bean gum (LBG), enriched with the tested BCAs, in controlling postharvest decays in artificially inoculated mandarins. The population dynamics of BCAs on wounds and the magnitude of peroxidase (POD) and superoxide dismutase (SOD) in fruit tissues were also investigated after treatments of mandarins with antagonistic yeasts. W. anomalus BS91, M. pulcherrima MPR3 and A. pullulans PI1 provided excellent control of postharvest decays caused by P. digitatum and P. italicum on mandarins, both when the yeasts were used alone and in combination with LBG, which enhanced the yeast cell viability over time. Finally, the increased activity of POD and lower decrease in SOD activity in response to BCAs application in mandarin fruits confirmed their involvement in the biocontrol mechanism. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. The major volatile compound 2-phenylethanol from the biocontrol yeast, Pichia anomala, inhibits growth and expression of aflatoxin biosynthetic genes of Aspergillus flavus.

    PubMed

    Hua, Sui Sheng T; Beck, John J; Sarreal, Siov Bouy L; Gee, Wai

    2014-05-01

    Aspergillus flavus is a ubiquitous saprophyte that is able to produce the most potent natural carcinogenic compound known as aflatoxin B1 (AFB1). This toxin frequently contaminates crops including corn, cotton, peanuts, and tree nuts causing substantial economic loss worldwide. Consequently, more than 100 countries have strict regulations limiting AFB1 in foodstuffs and feedstuffs. Plants and microbes are able to produce volatile compounds that act as a defense mechanism against other organisms. Pichia anomala strain WRL-076 is a biocontrol yeast currently being tested to reduce AF contamination of tree nuts in California. We used the SPME-GC/MS analysis and identified the major volatile compound produced by this strain to be 2-phenylethanol (2-PE). It inhibited spore germination and AF production of A. flavus. Inhibition of AF formation by 2-PE was correlated with significant down regulation of clustering AF biosynthesis genes as evidenced by several to greater than 10,000-fold decrease in gene expression. In a time-course analysis we found that 2-PE also altered the expression patterns of chromatin modifying genes, MYST1, MYST2, MYST3, gcn5, hdaA and rpdA. The biocontrol capacity of P. anomala can be attributed to the production of 2-PE, which affects spore germination, growth, toxin production, and gene expression in A. flavus.

  1. Multifunctional exopolysaccharides from Pseudomonas aeruginosa PF23 involved in plant growth stimulation, biocontrol and stress amelioration in sunflower under saline conditions.

    PubMed

    Tewari, Sakshi; Arora, Naveen Kumar

    2014-10-01

    Isolate PF23 selected from among 110 fluorescent pseudomonads, identified as Pseudomonas aeruginosa, displayed salinity tolerance and exopolysaccharides (EPS) production up to 2,000 mM NaCl concentration. EPS-defective mutant PF23(EPS-) of the isolate showed 86 % reduction in EPS production in comparison with wild strain. Defect in EPS production brought loss in salt tolerance capability. Purified EPS obtained from PF23 displayed multiple roles. At low concentration EPS functioned as biocontrol agent, at high concentration EPS behaved as osmoprotective or stress ameliorating metabolite and when introduced in saline soil, served as a plant growth promotor along with seed biopriming agent. Both in planta and in vivo studies were performed taking sunflower as a test crop and it was observed that PF23 showed plant growth promotion and significant biocontrol potential against dreadful phytopathogen Macrophomina phaseolina (under saline conditions). The mutant PF23(EPS-) was ineffective under saline conditions both in growth enhancement as well as in disease suppression. The study reports a potent strain, Pseudomonas aeruginosa PF23, capable of enhancing production of sunflower crop in semiarid regions and minimizing the incidence of charcoal rot disease in sunflower.

  2. Mutation of rpiA in Enterobacter cloacae decreases seed and root colonization and biocontrol of damping-off caused by Pythium ultimum on cucumber.

    PubMed

    Lohrke, Scott M; Dery, Pierre D; Li, Wei; Reedy, Ralph; Kobayashi, Donald Y; Roberts, Daniel R

    2002-08-01

    Strains of Enterobacter cloacae show promise as biocontrol agents for Pythium ultimum-induced damping-off on cucumber and other crops. E. cloacae A145 is a mini-Tn5 Km transposon mutant of strain 501R3 that was significantly reduced in suppression of damping-off on cucumber caused by P. ultimum. Strain A145 was deficient in colonization of cucumber, sunflower, and wheat seeds and significantly reduced in colonization of corn and cowpea seeds relative to strain 501R3. Populations of strain A145 were also significantly lower than those of strain 501R3 at all sampling times in cucumber, wheat, and sunflower rhizosphere. Populations of strain A145 were not detectable in any rhizosphere after 42 days, while populations of strain 501R3 remained at substantial levels throughout all experiments. Molecular characterization of strain A145 indicated mini-Tn5 Km was inserted in a region of the E. cloacae genome with a high degree of DNA and amino acid sequence similarity to rpiA, which encodes ribose-5-phosphate isomerase. In Escherichia coli, RpiA catalyzes the interconversion of ribose-5-phosphate and ribulose-5-phosphate and is a key enzyme in the pentose phosphate pathway. Ribose-5-phosphate isomerase activity in cell lysates from strain A145 was approximately 3.5% of that from strain 501R3. In addition, strain A145 was a ribose auxotroph, as expected for an rpiA mutant. Introduction of a 1.0-kb DNA fragment containing only the rpiA homologue into strain A145 restored ribose phosphate isomerase activity, prototrophy, seedling colonization, and disease suppression to levels similar to those associated with strain 501R3. Experiments reported here indicate a key role for rpiA and possibly the pentose phosphate pathway in suppression of damping-off and colonization of subterranean portions of plants by E. cloacae.

  3. Whole-Genome Shotgun Sequence of Bacillus mojavensis Strain RRC101, an Endophytic Bacterium Antagonistic to the Mycotoxigenic Endophytic Fungus Fusrium verticillioides

    USDA-ARS?s Scientific Manuscript database

    Here we report the whole genome shotgun sequence of Bacillus mojavensis strain RRC101, isolated from a maize kernel. This strain is antagonistic to the mycotoxigenic plant pathogen Fusarium verticillioides, and grows within maize tissue, suggesting potential as an endophytic biocontrol agent....

  4. Heterologous expression of VHb can improve the yield and quality of biocontrol fungus Paecilomyces lilacinus, during submerged fermentation.

    PubMed

    Zhang, Shumeng; Wang, Jieping; Wei, Yale; Tang, Qing; Ali, Maria Kanwal; He, Jin

    2014-10-10

    Paecilomyces lilacinus is an egg-parasitic fungus which is effective against plant-parasitic nematodes and it has been successfully commercialized for the control of many plant-parasitic nematodes. However, during the large-scale industrial fermentation process of the filamentous fungus, the dissolved oxygen supply is a limiting factor, which influences yield, product quality and production cost. To solve this problem, we intended to heterologously express VHb in P. lilacinus ACSS. After optimizing the vgb gene, we fused it with a selection marker gene nptII, a promoter PgpdA and a terminator TtrpC. The complete expression cassette PgpdA-nptII-vgb-TtrpC was transferred into P. lilacinus ACSS by Agrobacterium tumefaciens-mediated transformation. Consequently, we successfully screened an applicable fungus strain PNVT8 which efficiently expressed VHb. The submerged fermentation experiments demonstrated that the expression of VHb not only increased the production traits of P. lilacinus such as biomass and spore production, but also improved the beneficial product quality and application value, due to the secretion of more protease and chitinase. It can be speculated that the recombinant strain harboring vgb gene will have a growth advantage over the original strain under anaerobic conditions in soil and therefore will possess higher biocontrol efficiency against plant-parasitic nematodes.

  5. Biocontrol of Postharvest Anthracnose of Mango Fruit with Debaryomyces Nepalensis and Effects on Storage Quality and Postharvest Physiology.

    PubMed

    Luo, Shanshan; Wan, Bin; Feng, Shuhan; Shao, Yuanzhi

    2015-11-01

    Anthracnose is presently recognized as one of the most important postharvest disease of mango worldwide. To control the disease, chemical fungicides for a long time was widely used among fruit farmers, but recently found that pathogen had developed increasingly resistance to it. With people's growing desire of healthy and green food, finding new and environmentally friendly biological control approach was very necessary. In this paper, we provided a kind of new antagonistic yeast which enriched the strain resources and the efficacy of Debaryomyces nepalensis against postharvest anthracnose of mango fruit and the influence on quality parameters were investigated. The results showed that the decay incidence and lesion diameter of postharvest anthracnose of mango treated by D. nepalensis were significantly reduced compared with the control fruit stored at 25 °C for 30 d or at 15 °C for 40 d, and the higher concentration of D. nepalensis was, the better the efficacy of the biocontrol was. Study also found that 1 h was the best treatment duration and antagonistic yeast inoculated earlier had good biocontrol effect on anthracnose. Meanwhile, treatment by D. nepalensis could significantly reduce postharvest anthracnose of mango, delay the decrease in firmness, TSS, TA, and ascorbic acid value, and do not impair surface color during postharvest storage. Moreover, the increase in MDA (malondialdehyde) content and increase in cell membrane permeability of fruit treated by D. nepalensis was highly inhibited. The results suggested D. nepalensis treatment could not only maintain storage quality of mango fruit, but also decrease the decay incidence to anthracnose disease. All these results indicated that D. nepalensis has great potential for development of commercial formulations to control postharvest pathogens of mango fruit.

  6. Impact of Biocontrol Pseudomonas fluorescens CHA0 and a Genetically Modified Derivative on the Diversity of Culturable Fungi in the Cucumber Rhizosphere

    PubMed Central

    Girlanda, M.; Perotto, S.; Moenne-Loccoz, Y.; Bergero, R.; Lazzari, A.; Defago, G.; Bonfante, P.; Luppi, A. M.

    2001-01-01

    Little is known about the effects of Pseudomonas biocontrol inoculants on nontarget rhizosphere fungi. This issue was addressed using the biocontrol agent Pseudomonas fluorescens CHA0-Rif, which produces the antimicrobial polyketides 2,4-diacetylphloroglucinol (Phl) and pyoluteorin (Plt) and protects cucumber from several fungal pathogens, including Pythium spp., as well as the genetically modified derivative CHA0-Rif(pME3424). Strain CHA0-Rif(pME3424) overproduces Phl and Plt and displays improved biocontrol efficacy compared with CHA0-Rif. Cucumber was grown repeatedly in the same soil, which was left uninoculated, was inoculated with CHA0-Rif or CHA0-Rif(pME3424), or was treated with the fungicide metalaxyl (Ridomil). Treatments were applied to soil at the start of each 32-day-long cucumber growth cycle, and their effects on the diversity of the rhizosphere populations of culturable fungi were assessed at the end of the first and fifth cycles. Over 11,000 colonies were studied and assigned to 105 fungal species (plus several sterile morphotypes). The most frequently isolated fungal species (mainly belonging to the genera Paecilomyces, Phialocephala, Fusarium, Gliocladium, Penicillium, Mortierella, Verticillium, Trichoderma, Staphylotrichum, Coniothyrium, Cylindrocarpon, Myrothecium, and Monocillium) were common in the four treatments, and no fungal species was totally suppressed or found exclusively following one particular treatment. However, in each of the two growth cycles studied, significant differences were found between treatments (e.g., between the control and the other treatments and/or between the two inoculation treatments) using discriminant analysis. Despite these differences in the composition and/or relative abundance of species in the fungal community, treatments had no effect on species diversity indices, and species abundance distributions fit the truncated lognormal function in most cases. In addition, the impact of treatments at the 32-day

  7. The SRAP based molecular diversity related to antifungal and antioxidant bioactive constituents for biocontrol potentials of Trichoderma against Sclerotium rolfsii Scc.

    PubMed

    Hirpara, Darshna G; Gajera, H P; Bhimani, R D; Golakiya, B A

    2016-08-01

    The study was performed to examine 11 isolates of Trichoderma for their bio-control potentials against Sclerotium rolfsii Sacc. causing stem rot in groundnut. The antagonists Trichoderma were subjected to sequence related amplified polymorphism (SRAP) based molecular diversity analysis and compared with their hardness to S. rolfsii with respect to secretary antifungal and antioxidant profile. T. virens NBAII Tvs 12 evident highest (87.91 %) growth inhibition of test pathogen followed by T. koningii MTCC 796 (67.03 %) at 7 days after inoculation (DAI). Microscopic study confirmed biocontrol mechanism as mycoparasitism for Tvs 12 and antibiosis for MTCC 796. The growth inhibition of test pathogen was significantly negatively correlated with sclerotia formation and lipid peroxidation during antagonism due to release of secretary bioactive antioxidants by antagonists to terminate oxidative burst generated by S. rolfsii and causing inhibition of sclerotium formation. The GC-MS profile identified antifungal and antioxidant constituents hexadecane, 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester, 1-hexadecanesulfonyl chloride, and octadecane in potent antagonists Tvs 12; and nonacosane and octadecane in MTCC 796 along with two novel compounds 1-pentadecene and 1-heneicosyl formate for biocontrol activity. Molecular diversity of Trichoderma isolates associated with antagonistic activity was assessed by SRAP markers. The 115 primer combinations generate total 1328 amplified products of which, 1095 are shared polymorphic and 199 are unique polymorphic. The 15 SRAP combinations produced 18 bands to diagnose best antagonist Tvs 12 and 13 SRAP combinations generated 19 unique bands for identification of MTCC 796. The mycoparasitic antagonist Tvs 12 would be the best antagonist and released unique antifungal and antioxidant constituents to combat pathogen infection. The SRAP based genetic diversity indicates Tvs12 strain clustered with T. viride NBAII Tv23 and shared

  8. Genomic and metabolic traits endow Bacillus velezensis CC09 with a potential biocontrol agent in control of wheat powdery mildew disease.

    PubMed

    Cai, Xun-Chao; Liu, Chang-Hong; Wang, Bao-Tong; Xue, Ya-Rong

    2017-03-01

    Bacillus velezensis CC09, which was isolated from healthy leaves of Cinnamomum camphora and previously identified as Bacillus amyloliquefaciens CC09, shows great potential as a new biocontrol agent, in control of many phytopathogenic diseases. To extend our understanding of the potential antifungal capacities, we did a whole genome analysis of strain CC09. Result shows that strain CC09 has a relatively large genome size (4.17Mb) with an average GC content of 46.1%, and 4021 predicted genes. Thirteen secondary metabolites encoding clusters have been identified within the genome of B. velezensis CC09 using genome mining technique. Data of comparative genomic analysis indicated that 3 of the clusters are conserved by all strains of B. velezensis, B. amyloliquefaciens and B. subtilis 168, 9 by B. velezensis and B. amyloliquefaciens, and 2 by all strains of B. velezensis. Another 2 clusters encoding NRPS (Non-Ribosomal Peptide Synthetases) and NRPS-TransATPKS (NRPS and trans-Acyl Transferase Polyketide Synthetases) respectively are observed only in 15 B. velezensis strains, which might lead to the synthesis of novel bioactive compounds and could be explored as antimicrobial agents in the future. These clusters endow B. velezensis CC09 with strong and broad antimicrobial activities, for example, in control of wheat powdery mildew disease. Moreover, our data further confirmed the taxonomy of strain CC09 is a member of B. velezensis rather than a strain of B. amyloliquefaciens based on core genome sequence analysis using phylogenomic approach.

  9. The Potential of Streptomyces as Biocontrol Agents against the Rice Blast Fungus, Magnaporthe oryzae (Pyricularia oryzae)

    PubMed Central

    Law, Jodi Woan-Fei; Ser, Hooi-Leng; Khan, Tahir M.; Chuah, Lay-Hong; Pusparajah, Priyia; Chan, Kok-Gan; Goh, Bey-Hing; Lee, Learn-Han

    2017-01-01

    Rice is a staple food source for more than three billion people worldwide. However, rice is vulnerable to diseases, the most destructive among them being rice blast, which is caused by the fungus Magnaporthe oryzae (anamorph Pyricularia oryzae). This fungus attacks rice plants at all stages of development, causing annual losses of approximately 10–30% in various rice producing regions. Synthetic fungicides are often able to effectively control plant diseases, but some fungicides result in serious environmental and health problems. Therefore, there is growing interest in discovering and developing new, improved fungicides based on natural products as well as introducing alternative measures such as biocontrol agents to manage plant diseases. Streptomyces bacteria appear to be promising biocontrol agents against a wide range of phytopathogenic fungi, which is not surprising given their ability to produce various bioactive compounds. This review provides insight into the biocontrol potential of Streptomyces against the rice blast fungus, M. oryzae. The ability of various Streptomyces spp. to act as biocontrol agents of rice blast disease has been studied by researchers under both laboratory and greenhouse/growth chamber conditions. Laboratory studies have shown that Streptomyces exhibit inhibitory activity against M. oryzae. In greenhouse studies, infected rice seedlings treated with Streptomyces resulted in up to 88.3% disease reduction of rice blast. Studies clearly show that Streptomyces spp. have the potential to be used as highly effective biocontrol agents against rice blast disease; however, the efficacy of any biocontrol agent may be affected by several factors including environmental conditions and methods of application. In order to fully exploit their potential, further studies on the isolation, formulation and application methods of Streptomyces along with field experiments are required to establish them as effective biocontrol agents. PMID:28144236

  10. The Potential of Streptomyces as Biocontrol Agents against the Rice Blast Fungus, Magnaporthe oryzae (Pyricularia oryzae).

    PubMed

    Law, Jodi Woan-Fei; Ser, Hooi-Leng; Khan, Tahir M; Chuah, Lay-Hong; Pusparajah, Priyia; Chan, Kok-Gan; Goh, Bey-Hing; Lee, Learn-Han

    2017-01-01

    Rice is a staple food source for more than three billion people worldwide. However, rice is vulnerable to diseases, the most destructive among them being rice blast, which is caused by the fungus Magnaporthe oryzae (anamorph Pyricularia oryzae). This fungus attacks rice plants at all stages of development, causing annual losses of approximately 10-30% in various rice producing regions. Synthetic fungicides are often able to effectively control plant diseases, but some fungicides result in serious environmental and health problems. Therefore, there is growing interest in discovering and developing new, improved fungicides based on natural products as well as introducing alternative measures such as biocontrol agents to manage plant diseases. Streptomyces bacteria appear to be promising biocontrol agents against a wide range of phytopathogenic fungi, which is not surprising given their ability to produce various bioactive compounds. This review provides insight into the biocontrol potential of Streptomyces against the rice blast fungus, M. oryzae. The ability of various Streptomyces spp. to act as biocontrol agents of rice blast disease has been studied by researchers under both laboratory and greenhouse/growth chamber conditions. Laboratory studies have shown that Streptomyces exhibit inhibitory activity against M. oryzae. In greenhouse studies, infected rice seedlings treated with Streptomyces resulted in up to 88.3% disease reduction of rice blast. Studies clearly show that Streptomyces spp. have the potential to be used as highly effective biocontrol agents against rice blast disease; however, the efficacy of any biocontrol agent may be affected by several factors including environmental conditions and methods of application. In order to fully exploit their potential, further studies on the isolation, formulation and application methods of Streptomyces along with field experiments are required to establish them as effective biocontrol agents.

  11. Indirect effects of biocontrol of an invasive riparian plant (Tamarix) alters habitat and reduces herpetofauna abundance

    USGS Publications Warehouse

    Bateman, H.L.; Merritt, D.M.; Glenn, E.P.; Nagler, P.L.

    2014-01-01

    The biological control agent (tamarisk leaf beetle, Diorhabda spp.) is actively being used to defoliate exotic saltcedar or tamarisk (Tamarix spp.) in riparian ecosystems in western USA. The Virgin River in Arizona and Nevada is a system where tamarisk leaf beetle populations are spreading. Saltcedar biocontrol, like other control methods, has the potential to affect non-target species. Because amphibians and reptiles respond to vegetation changes in habitat and forage in areas where beetles are active, herpetofauna are model taxa to investigate potential impacts of biocontrol defoliation. Our objectives related herpetofauna abundance to vegetation cover and indices (normalized difference vegetation index, NDVI; enhanced vegetation index, EVI) and timing of biocontrol defoliation. We captured herpetofauna and ground-dwelling arthropods in trap arrays and measured vegetation using remotely sensed images and on-the-ground measurements at 16–21 sites 2 years before (2009–2010) and 2 years following (2011–2012) biocontrol defoliation. Following defoliation, riparian stands (including stands mixed with native and exotic trees and stands of monotypic exotic saltcedar) had significantly lower NDVI and EVI values and fewer captures of marked lizards. Total captures of herpetofauna (toads, lizards, and snakes) were related to higher vegetation cover and sites with a lower proportion of saltcedar. Our results suggest that effects of biocontrol defoliation are likely to be site-specific and depend upon the proportion of native riparian trees established prior to biocontrol introduction and defoliation. The mechanisms by which habitat structure, microclimate, and ultimately vertebrate species are affected by exotic plant biocontrol riparian areas should be a focus of natural-resource managers.

  12. Biocontrol of collar rot disease of betelvine (Piper betle L.) caused by Sclerotium rolfsii by using rhizosphere-competent Pseudomonas fluorescens NBRI-N6 and P. fluorescens NBRI-N.

    PubMed

    Singh, Anand; Mehta, Sangeeta; Singh, Harikesh Bahadur; Nautiyal, Chandra Shekhar

    2003-08-01

    Collar rot disease of betelvine (Piper betle L.) caused by Sclerotium rolfsii is difficult to control by conventional means by use of chemicals; therefore, use of biocontrol agents is desirable. In the present study, 186 bacterial strains of different morphological types were screened for their biocontrol activity against S. rolfsii under in vitro conditions. Two strains, Pseudomonas fluorescens NBRI-N6 and P. fluorescens NBRI-N, were selected for further studies because of their ability to inhibit the mycelial growth of the pathogen significantly. Spontaneous rifampicin-resistant (Rif) derivatives of P. fluorescens NBRI-N6 and P. fluorescens NBRI-N showing growth rate and membrane protein composition comparable to the wild type were selected to facilitate their monitoring in the rhizosphere. Field trials demonstrated that strain P. fluorescens NBRI-N6 was better than P. fluorescens NBRI-N in increasing the yield of betelvine significantly, whereas a consortium of the two strains controlled the disease more than either of the strains. The screening method should prove useful in identifying rhizosphere bacteria with the greatest potential for controlling diseases caused by phytopathogenic fungi.

  13. Use of AFLP for differentiation of Metschnikowia pulcherrima strains for postharvest disease biological control.

    PubMed

    Spadaro, D; Sabetta, W; Acquadro, A; Portis, E; Garibaldi, A; Gullino, M L

    2008-01-01

    Metschnikowia pulcherrima occurs naturally on fruits, buds and floral parts of apple trees. Some strains are effective as biocontrol agents against postharvest decay of apples and other fruits. The usefulness of the amplified fragment length polymorphism (AFLP) technique was evaluated for the genetic analysis of 26 strains of M. pulcherrima, isolated from different sources in different geographical regions. With six AFLP primer pairs, 729 polymorphic bands were scored. The technique showed a high discriminatory power. Genetic relationships between strains were also estimated using AFLP. All the isolates from the carposphere of apple, previously tested as biocontrol agents, were grouped in a single cluster with a high bootstrap value (97), indicating robustness and reproducibility. AFLP patterns could clearly distinguish the different strains and research is in progress to use some putative specific bands for single tag sequence (STS) conversion to develop isolate-specific markers.

  14. Biocontrol of larval mosquitoes by Acilius sulcatus (Coleoptera: Dytiscidae)

    PubMed Central

    Chandra, Goutam; Mandal, Samir K; Ghosh, Arup K; Das, Dipanwita; Banerjee, Siddhartha S; Chakraborty, Sumanta

    2008-01-01

    Background Problems associated with resistant mosquitoes and the effects on non-target species by chemicals, evoke a reason to find alternative methods to control mosquitoes, like the use of natural predators. In this regard, aquatic coleopterans have been explored less compared to other insect predators. In the present study, an evaluation of the role of the larvae of Acilius sulcatus Linnaeus 1758 (Coleoptera: Dytiscidae) as predator of mosquito immatures was made in the laboratory. Its efficacy under field condition was also determined to emphasize its potential as bio-control agent of mosquitoes. Methods In the laboratory, the predation potential of the larvae of A. sulcatus was assessed using the larvae of Culex quinquefasciatus Say 1823 (Diptera: Culicidae) as prey at varying predator and prey densities and available space. Under field conditions, the effectiveness of the larvae of A. sulcatus was evaluated through augmentative release in ten cemented tanks hosting immatures of different mosquito species at varying density. The dip density changes in the mosquito immatures were used as indicator for the effectiveness of A. sulcatus larvae. Results A single larva of A. sulcatus consumed on an average 34 IV instar larvae of Cx. quinquefasciatus in a 24 h period. It was observed that feeding rate of A. sulcatus did not differ between the light-on (6 a.m. – 6 p.m.), and dark (6 p.m. – 6 a.m.) phases, but decreased with the volume of water i.e., space availability. The prey consumption of the larvae of A. sulcatus differed significantly (P < 0.05) with different prey, predator and volume combinations, revealed through univariate ANOVA. The field study revealed a significant decrease (p < 0.05) in larval density of different species of mosquitoes after 30 days from the introduction of A. sulcatus larvae, while with the withdrawal, a significant increase (p < 0.05) in larval density was noted indicating the efficacy of A. sulcatus in regulating mosquito

  15. Biocontrol treatments confer protection against Verticillium dahliae infection of potato by inducing antimicrobial metabolites.

    PubMed

    El Hadrami, A; Adam, L R; Daayf, F

    2011-03-01

    Verticillium wilt, caused by Verticillium dahliae Kleb., is a serious potato (Solanum tuberosum L.) disease worldwide, and biocontrol represents a promising eco-friendly strategy to reduce its impact. We used extracts from Canada milk vetch (CMV) and a set of four V. dahliae-antagonistic bacterial strains to coat potato seeds at planting and examined the degree of protection provided against V. dahliae as well as accumulation of soluble phenolics as markers for induced resistance. All tested treatments were effective in reducing disease severity, and CMV showed the highest level of protection. In this treatment, flavonol-glycoside rutin was a highly abundant compound induced in potato tissues, with levels two to three times higher than those detected in noninoculated controls and V. dahliae-inoculated plants. We investigated dose-dependent effects of rutin on V. dahliae growth and sporulation in vitro and in planta. The effect of rutin on mycelial growth was inconsistent between disk assay and amended medium experiments. On the other hand, significant reduction of V. dahliae sporulation in vitro was consistently observed starting at 300 and 100 μM for isolates Vd-9 and Vd-21, respectively. We successfully detected 2-protocatechuoylphloroglucinolcarboxylic acid (2-PCPGCA) using ultra-performance liquid chromatography tandem mass spectrometry, indicating that V. dahliae dioxygenally oxidizes quercetin. Quercetin, as an aglycone, is freed from the sugar moiety by glucosidases and rhamnosidases produced by the fungus and is a substrate for quercetinases. The occurrence of quercetinases in V. dahliae provides a background to formulate a hypothesis about how by-product 2-PCPGCA may be interfering with potato defenses.

  16. Bacteriophage biocontrol of Listeria monocytogenes on soft ripened white mold and red-smear cheeses

    PubMed Central

    Guenther, Susanne

    2011-01-01

    Soft-ripened cheeses belong to the type of food most often contaminated with Listeria monocytogenes, and they have been implicated in several outbreaks of listeriosis. Bacteriophages represent an attractive way to combat foodborne pathogens without affecting other properties of the food. We used the broad host range, virulent Listeria phage A511 for control of L. monocytogenes during the production and ripening phases of both types of soft-ripened cheeses, white mold (Camembert-type) cheese, as well as washed-rind cheese with a red-smear surface (Limburger-type). The surfaces of young, unripened cheese were inoculated with 101–103 cfu/cm2 L. monocytogenes strains Scott A (serovar 4b) or CNL 103/2005 (serovar 1/2a). Phage was applied at defined time points thereafter, in single or repeated treatments, at 3 × 108 or 1 × 109 pfu/cm2. With Scott A (103 cfu/cm2) and a single dose of A511 (3 × 108 pfu/cm2) on camembert-type cheese, viable counts dropped 2.5 logs at the end of the 21 day ripening period. Repeated phage application did not further inhibit the bacteria, whereas a single higher dose (1 × 109 pfu/cm2) was found to be more effective. On red-smear cheese ripened for 22 days, Listeria counts were down by more than 3 logs. Repeated application of A511 further delayed re-growth of Listeria, but did not affect bacterial counts after 22 days. With lower initial Listeria contamination (101–102 cfu/cm2), viable counts dropped below the limit of detection, corresponding to more than 6 logs reduction compared to the control. Our data clearly demonstrate the potential of bacteriophage for biocontrol of L. monocytogenes in soft cheese. PMID:22334865

  17. Leuconostoc citreum MB1 as biocontrol agent of Listeria monocytogenes in milk.

    PubMed

    Pujato, Silvina A; del L Quiberoni, Andrea; Candioti, Mario C; Reinheimer, Jorge A; Guglielmotti, Daniela M

    2014-05-01

    Cell-free supernatant from Leuconostoc citreum MB1 revealed specific antilisterial activity. Preliminary studies demonstrated the proteinaceous, heat-stable, bacteriocin-like trait of the antimicrobial components present in the supernatant. Determination of the genes encoding bacteriocins by PCR and DNA sequencing led to amplification products highly homologous with leucocin A (found in diverse Leuconostoc species) and UviB (found in Leuc. citreum KM20) sequences. Additionally, antimicrobial activity of cell-free supernatant from Leuc. citreum MB1 was revealed by an inhibition halo of the SDS-PAGE gel subjected to a direct detection using Listeria monocytogenes as indicator strain. Different assays were carried out to assess the capacity of Leuc.citreum MB1 to control List. monocytogenes growth: (i) inactivation kinetics of the pathogen by antilisterial compounds present in concentrated cell-free supernatant from Leuc. citreum MB1, (ii) evaluation of optimal Leuc. citreum MB1 initial concentration to obtain maximum List. monocytogenes ATCC 15313 inhibition, and (iii) biocontrol of List. monocytogenes ATCC 15313 with Leuc. citreum MB1 during growth in milk at refrigeration temperature. According to our results, it is unquestionable that at least one bacteriocin is active in Leuc. citreum MB1, since important antilisterial activity was verified either in its cell-free supernatant or in co-culture experiments. Co-culture tests showed that ∼107 CFU/ml Leuc. citreum MB1 was the optimal initial concentration to obtain maximum pathogen inhibition. Moreover, Leuc. citreum MB1 was able to delay List. monocytogenes growth at refrigerated temperature.

  18. Draft Genome Sequence of Burkholderia cordobensis Type Strain LMG 27620, Isolated from Agricultural Soils in Argentina

    PubMed Central

    Draghi, Walter Omar; Mancini Villagra, Ulises M.; Wall, Luis Gabriel

    2015-01-01

    Bacteria of the genus Burkholderia are commonly found in diverse ecological niches in nature. We report here the draft genome sequence of Burkholderia cordobensis type strain LMG 27620, isolated from agricultural soil in Córdoba, Argentina. This strain harbors several genes involved in chitin utilization and phenol degradation, which make it an interesting candidate for biocontrol purposes and xenobiotic degradation in polluted environments. PMID:26494680

  19. Novel role for reactive oxygen species (ROS) in host-antagonistic yeast-pathogen interactions in postharvest biocontrol systems

    USDA-ARS?s Scientific Manuscript database

    To achieve the full potential of postharvest biocontrol microorganisms as a viable commercial technology, fundamental knowledge on their mechanisms of action is crucial. After more than 20 years of postharvest biocontrol research, there is still limited understanding of interactions taking place be...

  20. Cost modeling of pseudomonoas fluorescens and pseudomonoas chlororphis biocontrol for competitive exclusion of salmonella enterica on tomatoes

    USDA-ARS?s Scientific Manuscript database

    Biocontrol measures may enhance postharvest interventions, however; published research on process-based models for biocontrol of foodborne pathogens on produce is limited. The aim of this research was to develop cost model estimates for competitive exclusion process using Pseudomonas fluorescens and...

  1. Exploring ComQXPA quorum-sensing diversity and biocontrol potential of Bacillus spp. isolates from tomato rhizoplane

    PubMed Central

    Oslizlo, A; Stefanic, P; Vatovec, S; Beigot Glaser, S; Rupnik, M; Mandic-Mulec, I

    2015-01-01

    Bacillus subtilis is a widespread and diverse bacterium t exhibits a remarkable intraspecific diversity of the ComQXPA quorum-sensing (QS) system. This manifests in the existence of distinct communication groups (pherotypes) that can efficiently communicate within a group, but not between groups. Similar QS diversity was also found in other bacterial species, and its ecological and evolutionary meaning is still being explored. Here we further address the ComQXPA QS diversity among isolates from the tomato rhizoplane, a natural habitat of B. subtilis, where these bacteria likely exist in their vegetative form. Because this QS system regulates production of anti-pathogenic and biofilm-inducing substances such as surfactins, knowledge on cell–cell communication of this bacterium within rhizoplane is also important from the biocontrol perspective. We confirm the presence of pherotype diversity within B. subtilis strains isolated from a rhizoplane of a single plant. We also show that B. subtilis rhizoplane isolates show a remarkable diversity of surfactin production and potential plant growth promoting traits. Finally, we discover that effects of surfactin deletion on biofilm formation can be strain specific and unexpected in the light of current knowledge on its role it this process. PMID:25757097

  2. Structure, chemistry, and biological activity of pseudophomins A and B, new cyclic lipodepsipeptides isolated from the biocontrol bacterium Pseudomonas fluorescens.

    PubMed

    Pedras, M Soledade C; Ismail, Nargis; Quail, J Wilson; Boyetchko, Susan M

    2003-04-01

    Pseudophomins A and B are cyclic lipodepsipeptides isolated from Pseudomonas fluorescens strain BRG100, a bacterium with potential application for biocontrol of plant pathogens and weeds. Their chemical structures were established by a combination of spectroscopic data, X-ray crystallography, and selective chemical degradation. This unique chemical degradation allowed the unambiguous determination of the absolute configuration of the amino acid residue Leu-1, due to gamma-lactam formation followed by selective cleavage of the adjacent N(8)-C(7) bond. To the best of our knowledge this is the first application of gamma-lactam formation to the determination of absolute configuration of an adjacent amino acid. Pseudophomin B showed higher antifungal activity against the phytopathogens Phoma lingam/Leptosphaeria maculans and Sclerotinia sclerotiorum than pseudophomin A, and is likely to be the main component responsible for the antifungal activity of EtOAc extracts of strain BRG100. By contrast, pseudophomin A showed stronger inhibition of green foxtail (Setaria viridis) root germination than pseudophomin B.

  3. Exploring ComQXPA quorum-sensing diversity and biocontrol potential of Bacillus spp. isolates from tomato rhizoplane.

    PubMed

    Oslizlo, A; Stefanic, P; Vatovec, S; Beigot Glaser, S; Rupnik, M; Mandic-Mulec, I

    2015-05-01

    Bacillus subtilis is a widespread and diverse bacterium t exhibits a remarkable intraspecific diversity of the ComQXPA quorum-sensing (QS) system. This manifests in the existence of distinct communication groups (pherotypes) that can efficiently communicate within a group, but not between groups. Similar QS diversity was also found in other bacterial species, and its ecological and evolutionary meaning is still being explored. Here we further address the ComQXPA QS diversity among isolates from the tomato rhizoplane, a natural habitat of B. subtilis, where these bacteria likely exist in their vegetative form. Because this QS system regulates production of anti-pathogenic and biofilm-inducing substances such as surfactins, knowledge on cell-cell communication of this bacterium within rhizoplane is also important from the biocontrol perspective. We confirm the presence of pherotype diversity within B. subtilis strains isolated from a rhizoplane of a single plant. We also show that B. subtilis rhizoplane isolates show a remarkable diversity of surfactin production and potential plant growth promoting traits. Finally, we discover that effects of surfactin deletion on biofilm formation can be strain specific and unexpected in the light of current knowledge on its role it this process.

  4. Thiamine-Auxotrophic Mutants of Pseudomonas fluorescens CHA0 Are Defective in Cell-Cell Signaling and Biocontrol Factor Expression

    PubMed Central

    Dubuis, Christophe; Rolli, Joëlle; Lutz, Matthias; Défago, Geneviève; Haas, Dieter

    2006-01-01

    In the biocontrol strain Pseudomonas fluorescens CHA0, the Gac/Rsm signal transduction pathway positively controls the synthesis of antifungal secondary metabolites and exoenzymes. In this way, the GacS/GacA two-component system determines the expression of three small regulatory RNAs (RsmX, RsmY, and RsmZ) in a process activated by the strain's own signal molecules, which are not related to N-acyl-homoserine lactones. Transposon Tn5 was used to isolate P. fluorescens CHA0 insertion mutants that expressed an rsmZ-gfp fusion at reduced levels. Five of these mutants were gacS negative, and in them the gacS mutation could be complemented for exoproduct and signal synthesis by the gacS wild-type allele. Furthermore, two thiamine-auxotrophic (thiC) mutants that exhibited decreased signal synthesis in the presence of 5 × 10−8 M thiamine were found. Under these conditions, a thiC mutant grew normally but showed reduced expression of the three small RNAs, the exoprotease AprA, and the antibiotic 2,4-diacetylphloroglucinol. In a gnotobiotic system, a thiC mutant was impaired for biological control of Pythium ultimum on cress. Addition of excess exogenous thiamine restored all deficiencies of the mutant. Thus, thiamine appears to be an important factor in the expression of biological control by P. fluorescens. PMID:16597964

  5. Genome analysis reveals insights of the endophytic Bacillus toyonensis BAC3151 as a potentially novel agent for biocontrol of plant pathogens.

    PubMed

    Lopes, Ralf; Cerdeira, Louise; Tavares, Grace S; Ruiz, Jeronimo C; Blom, Jochen; Horácio, Elvira C A; Mantovani, Hilário C; Queiroz, Marisa Vieira de

    2017-09-25

    Diseases caused by phytopathogenic microorganisms account for enormous losses for agribusiness. Although Bacillus species are recognized as being antimicrobial producers and some may provide benefits to plants, the association between Bacillus toyonensis and plants has not been studied. In this study, the whole-genome sequenced endophytic B. toyonensis BAC3151, which has demonstrated antimicrobial activity and quorum sensing inhibition of phytopathogenic bacteria, was investigated for its potential for the production of compounds for biocontrol of plant pathogens. Four whole-genome sequenced B. toyonensis strains shared 3811 protein-coding DNA sequences (CDSs), while strain-specific CDSs, such as biosynthetic gene clusters of antimicrobials, were associated with specific chromosomal regions and mobile genetic elements of the strains. B. toyonensis strains had a higher frequency of putative bacteriocins gene clusters than that of Bacillus species traditionally used for the production of antimicrobials. In addition, gene clusters potentially involved in the production of novel bacteriocins were found in BAC3151, as well as biosynthetic genes of several other compounds, including non-ribosomal peptides, N-acyl homoserine lactonase and chitinases, revealing a genetic repertoire for antimicrobial synthesis greater than that of other Bacillus strains that have demonstrated effective activity against phytopathogens. This study showed for the first time that B. toyonensis has potential to produce various antimicrobials, and the analyses performed indicated that the endophytic strain BAC3151 can be useful for the development of new strategies to control microbial diseases in plants that are responsible for large damages in agricultural crops.

  6. Ochratoxin A management in vineyards by Lobesia botrana biocontrol.

    PubMed

    Cozzi, Giuseppe; Somma, Stefania; Haidukowski, Miriam; Logrieco, Antonio F

    2013-01-02

    Grape berries attacked by Lobesia botrana larvae are more easily infected by Aspergillus section Nigri (black aspergilli) ochratoxigenic species. Two-year field trials were carried out in Apulia (Italy) to evaluate a bioinsecticide control strategy against L. botrana and the indirect effect on reducing ochratoxin A (OTA) contamination in vineyards. A commercial Bacillus thuringiensis formulate and an experimental Beauveria bassiana (ITEM-1559) formulate were tested in two vineyards cultivated with the same grape variety, Negroamaro, but with two different training systems (espalier and little-arbor techniques). In both years and training systems the treatments by B. bassiana ITEM-1559 significantly controlled L. botrana larvae attacks with effectiveness similar to B. thuringensis (more than 20%). A significant reduction of OTA concentrations (up to 80% compared to untreated controls) was observed only in the first year in both training systems, when the metereological parameters prior to harvest were more favorable to the insect attack. Results of field trials showed that B. bassiana ITEM-1559 is a valid bioinsecticide against L. botrana and that grape moth biocontrol is a strategy to reduce OTA contamination in vineyard in seasons with heavy natural infestation.

  7. Ochratoxin A Management in Vineyards by Lobesia botrana Biocontrol

    PubMed Central

    Cozzi, Giuseppe; Somma, Stefania; Haidukowski, Miriam; Logrieco, Antonio F.

    2013-01-01

    Grape berries attacked by Lobesia botrana larvae are more easily infected by Aspergillus section Nigri (black aspergilli) ochratoxigenic species. Two-year field trials were carried out in Apulia (Italy) to evaluate a bioinsecticide control strategy against L. botrana and the indirect effect on reducing ochratoxin A (OTA) contamination in vineyards. A commercial Bacillus thuringiensis formulate and an experimental Beauveria bassiana (ITEM-1559) formulate were tested in two vineyards cultivated with the same grape variety, Negroamaro, but with two different training systems (espalier and little-arbor techniques). In both years and training systems the treatments by B. bassiana ITEM-1559 significantly controlled L. botrana larvae attacks with effectiveness similar to B. thuringensis (more than 20%). A significant reduction of OTA concentrations (up to 80% compared to untreated controls) was observed only in the first year in both training systems, when the metereological parameters prior to harvest were more favorable to the insect attack. Results of field trials showed that B. bassiana ITEM-1559 is a valid bioinsecticide against L. botrana and that grape moth biocontrol is a strategy to reduce OTA contamination in vineyard in seasons with heavy natural infestation. PMID:23282670

  8. [The prospects of using transgenic insects in biocontrol programs].

    PubMed

    Tkachuk, A P; Kim, M V; Savitskiĭ, V Iu; Savitskiĭ, M Iu

    2011-01-01

    Methods of biocontrol are widely used for suppression of pests and human disease vectors. One of the key methods is insects sterilization (sterile insect technique--SIT), which currently is accomplished by irradiation. Radiation-exposed insects have reduced fitness so theis competitive abilities are diminished as compared to insects from wild populations. Modern bioengineering is capable of producing transgenic insects with predetermined traits, and by now the schemes for getting sterile insects without exposure to radiation are developed. Another area of modern studies is producing insects that are unable to transmit diseases malaria, for example. In the present review the implementation and perspectives are outlined for replacement of Anopheles wild populations with transgenic mosquitos. The main way for delivering the genetic material to recipient's genome is using transposon-based constructs. The markers of transgenesis are described. The potential danger for the environment of transgenic constructs remobilization and the necessity of their stabilization within the genome are emphasized. The existing methods of stabilization which involve the deletion of transposon terminal inverted repeats are described.

  9. Insight into tick biocontrol with special regard to fungi.

    PubMed

    Lonc, Elżbieta; Guz-Regner, Katarzyna; Kiewra, Dorota; Szczepańska, Anna

    2014-01-01

    The epidemiological and epizootic importance of ticks has been known for a few decades since of the discovery of their role as vectors of many new diseases, and the better detection of those already known. Given the durability of chemical preparations in the environment and the increasing problem of developing tick resistance, natural strategies for biological control are sought. A promising alternative to chemical pesticides is the use of entomopathogenic organisms for effective integrated pest management of low environmental impact. A number of promising microbes have been identified during the search for effective means of controlling the tick population, but the knowledge about the impact of these pathogens on the environment and other non-target organisms is still insufficient. Previous research has still not provided a definite answer about the safety of their use. It is known, however,that the chemicals which are currently used have a negative impact on the environment and/or cause resistance. No efficient biocompound has yet been devised for commercial use. Potential microorganisms for tick biocontrol (mainly bacteria and fungi) are natural tick pathogens, living in the same environment. With their adhesive properties, and their ability to digest the cuticle, they may constitute an appropriate ingredient of bioacaricides. Until now, fungal insecticides have been used only to control crop pests.

  10. Complete genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium of Calendula officinalis

    SciTech Connect

    Koeberl, Martina; White, Richard A.; Erschen, Sabine; Spanberger, Nora; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-13

    The genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium (PGPR) with broad-spectrum antagonistic activities against plant pathogenic fungi, bacteria and nematodes, consists of a single 3.9 Mb circular chromosome. The genome reveals genes putatively responsible for its promising biocontrol and PGP properties.

  11. Complete genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium of Calendula officinalis

    DOE PAGES

    Köberl, Martina; White, Richard A.; Erschen, Sabine; ...

    2015-08-13

    The genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium (PGPR) with broad-spectrum antagonistic activity against plant-pathogenic fungi, bacteria, and nematodes, consists of a single 3.9-Mb circular chromosome. The genome reveals genes putatively responsible for its promising biocontrol and PGP properties.

  12. Use of a Granular Bioplastic Formulation for Carrying Conidia of a Non-aflatoxigenic Strain of Aspergillus flavus

    USDA-ARS?s Scientific Manuscript database

    Previous research demonstrated that aflatoxin contamination in corn grown in Mississippi is reduced by field application of wheat grains pre-inoculated with the non-aflatoxigenic Aspergillus flavus strain NRRL 30797. To facilitate field applications of the biocontrol isolate, a series of laboratory ...

  13. Players over the Surface: Unraveling the Role of Exopolysaccharides in Zinc Biosorption by Fluorescent Pseudomonas Strain Psd

    PubMed Central

    Upadhyay, Anamika; Kochar, Mandira; Rajam, Manchikatla V.; Srivastava, Sheela

    2017-01-01

    Fluorescent Pseudomonas strain Psd is a soil isolate, possessing multiple plant growth promoting (PGP) properties and biocontrol potential. In addition, the strain also possesses high Zn2+ biosorption capability. In this study, we have investigated the role exopolysaccharides (EPS) play in Zn2+ biosorption. We have identified that alginates are the prime components contributing to Zn2+ biosorption. Deletion of the alg8 gene, which codes for a sub-unit of alginate polymerase, led to a significant reduction in EPS production by the organism. We have also demonstrated that the increased alginate production in response to Zn2+ exposure leads to improved biofilm formation by the strain. In the alg8 deletion mutant, however, biofilm formation was severely compromised. Further, we have studied the functional implications of Zn2+ biosorption by Pseudomonas strain Psd by demonstrating the effect on the PGP and biocontrol potential of the strain. PMID:28286498

  14. Myco-Biocontrol of Insect Pests: Factors Involved, Mechanism, and Regulation

    PubMed Central

    Sandhu, Sardul Singh; Sharma, Anil K.; Beniwal, Vikas; Goel, Gunjan; Batra, Priya; Kumar, Anil; Jaglan, Sundeep; Sharma, A. K.; Malhotra, Sonal

    2012-01-01

    The growing demand for reducing chemical inputs in agriculture and increased resistance to insecticides have provided great impetus to the development of alternative forms of insect-pest control. Myco-biocontrol offers an attractive alternative to the use of chemical pesticides. Myco-biocontrol agents are naturally occurring organisms which are perceived as less damaging to the environment. Their mode of action appears little complex which makes it highly unlikely that resistance could be developed to a biopesticide. Past research has shown some promise of the use of fungi as a selective pesticide. The current paper updates us about the recent progress in the field of myco-biocontrol of insect pests and their possible mechanism of action to further enhance our understanding about the biological control of insect pests. PMID:22567344

  15. Antibiosis by Pantoea agglomerans biocontrol strain E325 against Erwinia amylovora on apple blossom stigmas

    USDA-ARS?s Scientific Manuscript database

    Pantoea agglomerans E325, the active ingredient in a commercial product for fire blight control, was previously shown in vitro to produce a unique alkaline- and phosphate-sensitive antibiotic specific to Erwinia amylovora. Antibiosis was evaluated as a mode of antagonism on blossom stigmas using two...

  16. Essential oils from Algerian species of Mentha as new bio-control agents against phytopathogen strains.

    PubMed

    Benomari, Fatima Zahra; Andreu, Vanessa; Kotarba, Jules; Dib, Mohammed El Amine; Bertrand, Cédric; Muselli, Alain; Costa, Jean; Djabou, Nassim

    2017-09-02

    Chemical composition and antifungal activity of essential oils of Algerian Mentha species were studied. Chemical compositions of different Mentha species oils (Mentha rotundifolia, M. spicata, M. pulegium, and M. piperita) were investigated by capillary GC and GC/MS, and their antifungal activities were evaluated by means of paper disc diffusion method and minimum inhibitory concentration (MIC) assays. In total, 98 components from all Mentha species were identified. All oils were rich in monoterpene-oxygenated components. In addition, we reported fumigant antifungal activity of Algerian Mentha essential oils against four fungi: Botrytis cinerea, Penicillium expansum, Monilinia laxa, and M. fructigena. All oils demonstrated very good inhibition especially against B. cinerea, M. laxa, and M. fructigena. Both Monilinia fungi were extremely sensitive to all Algerian Mentha oils, which suggests that Mentha essential oils have the potential to be used as bio-pesticides to protect fruit trees, such as apple and pear trees, and provides an alternative to chemical pesticides.

  17. Characterization of strains of Bacillus mojavensis for biocontrol of a maize pathogen

    USDA-ARS?s Scientific Manuscript database

    The endophytic bacterium, Bacillus mojavensis controls fungal diseases in maize and other plants. The bacterium and its cultural extracts have been shown to be antagonistic to a pathogenic and mycotoxic fungus, Fusarium verticillioides. Extracts prepared from cultures of B. mojavensis contained an...

  18. Biosynthetic Mechanism for Sunscreens of the Biocontrol Agent Lysobacter enzymogenes

    PubMed Central

    Li, Yaoyao; Wang, Yansheng; Wang, Yulan; Wright, Stephen; Li, Yuezhong; Shen, Yuemao; Liu, Fengquan; Du, Liangcheng

    2013-01-01

    Lysobacter are ubiquitous environmental bacteria emerging as novel biocontrol agents and new sources of anti-infectives. So far, very little effort has been invested in the study of the biology of these Gram-negative gliding bacteria. Many Lysobacter species are characterized by their yellow-orange appearance. Using transposon mutagenesis, we identified a stand-alone polyketide synthase (PKS) gene cluster required for the pigment production in L. enzymogenes OH11. The yellow pigments were abolished in the “white” mutants generated by target-specific deletions of ketosynthase (KS), acyl carrier protein, or ketoreductase. Spectroscopic data suggested that the pigments belong to xanthomonadin-like aryl polyenes. Polyene-type polyketides are known to be biosynthesized by modular PKS (Type I), not by stand-alone PKS (Type II) which always contain the heterodimer KS-CLF (chain-length factor) as the key catalytic component. Remarkably, this aryl polyene PKS complex only contains the KS (ORF17), but not the CLF. Instead, a hypothetical protein (ORF16) is located immediately next to ORF17. ORF16–17 homologs are widespread in numerous uncharacterized microbial genomes, in which an ORF17 homolog is always accompanied by an ORF16 homolog. The deletion of ORF16 eliminated pigment production, and homology modeling suggested that ORF16 shares a structural similarity to the N-terminal half of CLF. A point-mutation of glutamine (Q166A) that is the conserved active site of known CLF abolished pigment production. The “white” mutants are significantly more sensitive to UV/visible light radiation or H2O2 treatment than the wild type. These results unveil the first example of Type II PKS-synthesized polyene pigments and show that the metabolites serve as Lysobacter “sunscreens” that are important for the survival of these ubiquitous environmental organisms. PMID:23826105

  19. Tick control: trapping, biocontrol, host management and other alternative strategies

    USGS Publications Warehouse

    Ginsberg, Howard S.; Edited by Sonenshine, Daniel E.; Roe, R. Michael

    2014-01-01

    Biology of Ticks is the most comprehensive work on tick biology and tick-borne diseases. This second edition is a multi-authored work, featuring the research and analyses of renowned experts across the globe. Spanning two volumes, the book examines the systematics, biology, structure, ecological adaptations, evolution, genomics and the molecular processes that underpin the growth, development and survival of these important disease-transmitting parasites. Also discussed is the remarkable array of diseases transmitted (or caused) by ticks, as well as modern methods for their control. This book should serve as a modern reference for students, scientists, physicians, veterinarians and other specialists. Volume I covers the biology of the tick and features chapters on tick systematics, tick life cycles, external and internal anatomy, and others dedicated to specific organ systems, specifically, the tick integument, mouthparts and digestive system, salivary glands, waste removal, salivary glands, respiratory system, circulatory system and hemolymph, fat body, the nervous and sensory systems and reproductive systems. Volume II includes chapters on the ecology of non-nidicolous and nidicolous ticks, genetics and genomics (including the genome of the Lyme disease vector Ixodes scapularis) and immunity, including host immune responses to tick feeding and tick-host interactions, as well as the tick's innate immune system that prevents and/or controls microbial infections. Six chapters cover in depth the many diseases caused by the major tick-borne pathogens, including tick-borne protozoa, viruses, rickettsiae of all types, other types of bacteria (e.g., the Lyme disease agent) and diseases related to tick paralytic agents and toxins. The remaining chapters are devoted to tick control using vaccines, acaricides, repellents, biocontrol, and, finally, techniques for breeding ticks in order to develop tick colonies for scientific study.

  20. Anolis lizards as biocontrol agents in mainland and island agroecosystems.

    PubMed

    Monagan, Ivan V; Morris, Jonathan R; Davis Rabosky, Alison R; Perfecto, Ivette; Vandermeer, John

    2017-04-01

    Our knowledge of ecological interactions that bolster ecosystem function and productivity has broad applications to the management of agricultural systems. Studies suggest that the presence of generalist predators in agricultural landscapes leads to a decrease in the abundance of herbivorous pests, but our understanding of how these interactions vary across taxa and along gradients of management intensity and eco-geographic space remains incomplete. In this study, we assessed the functional response and biocontrol potential of a highly ubiquitous insectivore (lizards in the genus Anolis) on the world's most important coffee pest, the coffee berry borer (Hypothalemus hampei). We conducted field surveys and laboratory experiments to examine the impact of land-use intensification on species richness and abundance of anoles and the capacity of anoles to reduce berry borer infestations in mainland and island coffee systems. Our results show that anoles significantly reduce coffee infestation rates in laboratory settings (Mexico, p = .03, F = 5.13 df = 1, 35; Puerto Rico, p = .014, F = 8.82, df = 1, 10) and are capable of consuming coffee berry borers in high abundance. Additionally, diversified agroecosystems bolster anole abundance, while high-intensity practices, including the reduction of vegetation complexity and the application of agrochemicals were associated with reduced anole abundance. The results of this study provide supporting evidence of the positive impact of generalist predators on the control of crop pests in agricultural landscapes, and the role of diversified agroecosystems in sustaining both functionally diverse communities and crop production in tropical agroecosystems.

  1. The ability of a cold-adapted Rhodotorula mucilaginosa strain from Tibet to control blue mold in pear fruit.

    PubMed

    Hu, Hao; Yan, Fujie; Wilson, Charles; Shen, Qing; Zheng, Xiaodong

    2015-12-01

    Cold-adapted yeasts were isolated from soil samples collected in Tibet and evaluated as potential biocontrol agents against blue mold (Penicillium expansum) of pear fruit in cold storage. YC1, an isolate identified as Rhodotorula mucilaginosa, was found to exhibit the greatest biocontrol activity among the different isolates that were screened. A washed cell suspension of YC1 exhibited the best biocontrol activity among three different preparations that were used in the current study. A concentration of 10(8) cells/ml reduced the incidence of decay to 35 %, compared to the control where decay incidence was 100 %. A higher intracellular level of trehalose and a higher proportion of polyunsaturated acids present in YC1, was associated with increased the tolerance of this strain to low temperatures, relative to the other strains that were evaluated. The increased tolerance to low temperature allowed the YC1 strain of yeast to more effectively compete for nutrients and space in wounded pear fruit that had been inoculated with spores of P. expansum and placed in cold storage. The present study demonstrated the ability to select cold-adapted yeasts from cold climates and use them as biocontrol agents of postharvest diseases of fruit placed in cold storage.

  2. Nutrient Effects on Biocontrol of Penicillium roqueforti by Pichia anomala J121 during Airtight Storage of Wheat

    PubMed Central

    Druvefors, Ulrika Ädel; Passoth, Volkmar; Schnürer, Johan

    2005-01-01

    The biocontrol yeast Pichia anomala inhibits the growth of a variety of mold species. We examined the mechanism underlying the inhibition of the grain spoilage mold Penicillium roqueforti by the biocontrol yeast P. anomala J121 during airtight storage. The biocontrol effect in a model grain silo with moist wheat (water activity of 0.96) was enhanced when complex medium, maltose, or glucose was added. Supplementation with additional nitrogen or vitamin sources did not affect the biocontrol activity of the yeast. The addition of complex medium or glucose did not significantly influence the yeast cell numbers in the silos, whether in the presence or absence of P. roqueforti. Mold growth was not influenced by the addition of nutrients, if cultivated without yeast. The products of glucose metabolism, mainly ethanol and ethyl acetate, increased after glucose addition to P. anomala-inoculated treatments. Our results suggest that neither competition for nutrients nor production of a glucose-repressible cell wall lytic enzyme is the main mode of action of biocontrol by P. anomala in this grain system. Instead, the mold-inhibiting effect probably is due to the antifungal action of metabolites, most likely a combination of ethyl acetate and ethanol, derived from glycolysis. The discovery that sugar amendments enhance the biocontrol effect of P. anomala suggests novel ways of formulating biocontrol yeasts. PMID:15812013

  3. Cytokinin production by Pseudomonas fluorescens G20-18 determines biocontrol activity against Pseudomonas syringae in Arabidopsis

    PubMed Central

    Großkinsky, Dominik K.; Tafner, Richard; Moreno, María V.; Stenglein, Sebastian A.; García de Salamone, Inés E.; Nelson, Louise M.; Novák, Ondřej; Strnad, Miroslav; van der Graaff, Eric; Roitsch, Thomas

    2016-01-01

    Plant beneficial microbes mediate biocontrol of diseases by interfering with pathogens or via strengthening the host. Although phytohormones, including cytokinins, are known to regulate plant development and physiology as well as plant immunity, their production by microorganisms has not been considered as a biocontrol mechanism. Here we identify the ability of Pseudomonas fluorescens G20-18 to efficiently control P. syringae infection in Arabidopsis, allowing maintenance of tissue integrity and ultimately biomass yield. Microbial cytokinin production was identified as a key determinant for this biocontrol effect on the hemibiotrophic bacterial pathogen. While cytokinin-deficient loss-of-function mutants of G20-18 exhibit impaired biocontrol, functional complementation with cytokinin biosynthetic genes restores cytokinin-mediated biocontrol, which is correlated with differential cytokinin levels in planta. Arabidopsis mutant analyses revealed the necessity of functional plant cytokinin perception and salicylic acid-dependent defence signalling for this biocontrol mechanism. These results demonstrate microbial cytokinin production as a novel microbe-based, hormone-mediated concept of biocontrol. This mechanism provides a basis to potentially develop novel, integrated plant protection strategies combining promotion of growth, a favourable physiological status and activation of fine-tuned direct defence and abiotic stress resilience. PMID:26984671

  4. Nutrient effects on biocontrol of Penicillium roqueforti by Pichia anomala J121 during airtight storage of wheat.

    PubMed

    Druvefors, Ulrika Adel; Passoth, Volkmar; Schnürer, Johan

    2005-04-01

    The biocontrol yeast Pichia anomala inhibits the growth of a variety of mold species. We examined the mechanism underlying the inhibition of the grain spoilage mold Penicillium roqueforti by the biocontrol yeast P. anomala J121 during airtight storage. The biocontrol effect in a model grain silo with moist wheat (water activity of 0.96) was enhanced when complex medium, maltose, or glucose was added. Supplementation with additional nitrogen or vitamin sources did not affect the biocontrol activity of the yeast. The addition of complex medium or glucose did not significantly influence the yeast cell numbers in the silos, whether in the presence or absence of P. roqueforti. Mold growth was not influenced by the addition of nutrients, if cultivated without yeast. The products of glucose metabolism, mainly ethanol and ethyl acetate, increased after glucose addition to P. anomala-inoculated treatments. Our results suggest that neither competition for nutrients nor production of a glucose-repressible cell wall lytic enzyme is the main mode of action of biocontrol by P. anomala in this grain system. Instead, the mold-inhibiting effect probably is due to the antifungal action of metabolites, most likely a combination of ethyl acetate and ethanol, derived from glycolysis. The discovery that sugar amendments enhance the biocontrol effect of P. anomala suggests novel ways of formulating biocontrol yeasts.

  5. Cytokinin production by Pseudomonas fluorescens G20-18 determines biocontrol activity against Pseudomonas syringae in Arabidopsis.

    PubMed

    Großkinsky, Dominik K; Tafner, Richard; Moreno, María V; Stenglein, Sebastian A; García de Salamone, Inés E; Nelson, Louise M; Novák, Ondřej; Strnad, Miroslav; van der Graaff, Eric; Roitsch, Thomas

    2016-03-17

    Plant beneficial microbes mediate biocontrol of diseases by interfering with pathogens or via strengthening the host. Although phytohormones, including cytokinins, are known to regulate plant development and physiology as well as plant immunity, their production by microorganisms has not been considered as a biocontrol mechanism. Here we identify the ability of Pseudomonas fluorescens G20-18 to efficiently control P. syringae infection in Arabidopsis, allowing maintenance of tissue integrity and ultimately biomass yield. Microbial cytokinin production was identified as a key determinant for this biocontrol effect on the hemibiotrophic bacterial pathogen. While cytokinin-deficient loss-of-function mutants of G20-18 exhibit impaired biocontrol, functional complementation with cytokinin biosynthetic genes restores cytokinin-mediated biocontrol, which is correlated with differential cytokinin levels in planta. Arabidopsis mutant analyses revealed the necessity of functional plant cytokinin perception and salicylic acid-dependent defence signalling for this biocontrol mechanism. These results demonstrate microbial cytokinin production as a novel microbe-based, hormone-mediated concept of biocontrol. This mechanism provides a basis to potentially develop novel, integrated plant protection strategies combining promotion of growth, a favourable physiological status and activation of fine-tuned direct defence and abiotic stress resilience.

  6. Functional and structural microbial diversity in organic and conventional viticulture: organic farming benefits natural biocontrol agents.

    PubMed

    Schmid, Florian; Moser, Gerit; Müller, Henry; Berg, Gabriele

    2011-03-01

    Statistically significant differences in the structure and function of above-ground grapevine-associated microorganisms from organically and conventionally managed vineyards were found. Aureobasidium pullulans, a copper-detoxifying fungus and biocontrol agent, plays a key role in explaining these differences. The black fungus was strongly enriched in the communities of organically managed plants and yielded a higher indigenous antiphytopathogenic potential.

  7. Stress tolerance and biocontrol performance of the yeast antagonist, Candida diversa, change with morphology transition.

    PubMed

    Li, Guangkun; Chi, Mengshan; Chen, Huizhen; Sui, Yuan; Li, Yan; Liu, Yongsheng; Zhang, Xiaojing; Sun, Zhiqiang; Liu, Guoqing; Wang, Qi; Liu, Jia

    2016-02-01

    As an eco-friendly management method, biological control of postharvest diseases, utilizing antagonistic yeasts, is a research topic receiving considerable attention. Detailed knowledge on the biology of yeast antagonists is crucial when considering their potential application and development as biocontrol products. Changes in the growth form, such as single-cell to pseudohyphae, have been associated with the mode of action in postharvest biocontrol yeasts. In this study, the antagonistic yeast, Candida diversa, reversibly shifted from a single-cell morphology on yeast peptone dextrose (YPD) medium with 2 % agar to a pseudohyphal morphology on YPD with 0.3 % agar. The tolerance of the pseudohyphal form to heat and oxidative stresses, as well as the biocontrol efficacy against Botrytis cinerea on apple and kiwifruit stored at 25 and 4 °C, was significantly higher as compared to the single-cell form. This study provides new information on the ability of C. diversa to change its morphology and the impact of the morphology shift on stress tolerance and biocontrol performance.

  8. [Effect of adaptive biocontrol sessions in the presence of strong professional skill].

    PubMed

    Antonov, A A; Ershova, T A; Dudukin, A V

    2011-01-01

    Effect of adaptive biocontrol on efficiency of the operator possessing stable skills in simulating manual control of Soyuz hang-up and berthing to the International space station (ISS) was studied. Job quality was evaluated by the time, energy and precision parameters. The investigation used an ISS computer model and mock-ups of the board operating controls.

  9. Miconia biocontrol: Where are we going and when will we get there?

    Treesearch

    M.T. Johnson

    2010-01-01

    We have made much progress in evaluating potential agents for biocontrol of miconia, and several appear likely to be suitable for future introduction to Hawaiÿi. Unfortunately, none of them is an obvious silver bullet. We face the challenge of prioritizing the existing candidates and inventing the combination of agents that will achieve our goals. Now is an opportune...

  10. Yeast Biocontrol of a Fungal Plant Disease: A Model for Studying Organism Interrelationships

    ERIC Educational Resources Information Center

    Chanchaichaovivat, Arun; Panijpan, Bhinyo; Ruenwongsa, Pintip

    2008-01-01

    An experiment on the action of the yeast, "Saccharomyces cerevisiae", against a fungal plant disease is proposed for secondary students (Grade 11) to support their study of organism interrelationship. This biocontrol experiment serves as the basis for discussing relationships among three organisms (red chilli fruit, "Saccharomyces cerevisiae," and…

  11. Linking habitat management and conservation biocontrol through gut-content analysis

    USDA-ARS?s Scientific Manuscript database

    Increasing the habitat diversity of agricultural fields can lead to more effective biocontrol of arthropod pests. Annual cropping systems are exposed to frequent disturbance and lack habitat diversity; therefore it is important to develop strategies that can improve ecosystem services such as bioco...

  12. What magnitude are observed non-target impacts from weed biocontrol?

    USDA-ARS?s Scientific Manuscript database

    A systematic review focused by plant on non-target impacts from agents deliberately introduced for the biological control of weeds found significant non-target impacts to be rare. The magnitude of direct impact of 43 biocontrol agents on 140 non-target plants was retrospectively categorized using a ...

  13. Antagonistic activity of Bacillus subtilis SB1 and its biocontrol effect on tomato bacterial wilt

    USDA-ARS?s Scientific Manuscript database

    A potential biocontrol agent of bacterial wilt, Bacillus subtilis SB1, isolated from tomato roots, showed a broad-spectrum of antimicrobial activity in in vitro experiments. It inhibited the growth of many plant pathogens, including Ralstonia solanacearum, Xanthomonas oryzae pv. oryzae, Fusarium ox...

  14. Agar composition affects in vitro screening of biocontrol activity of antagonistic microorganisms.

    PubMed

    Bosmans, L; De Bruijn, I; De Mot, R; Rediers, H; Lievens, B

    2016-08-01

    Agar-based screening assays are the method of choice when evaluating antagonistic potential of bacterial biocontrol-candidates against pathogens. We showed that when using the same medium, but different agar compositions, the activity of a bacterial antagonist against Agrobacterium was strongly affected. Consequently, results from in vitro screenings should be interpreted cautiously. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Yeast Biocontrol of a Fungal Plant Disease: A Model for Studying Organism Interrelationships

    ERIC Educational Resources Information Center

    Chanchaichaovivat, Arun; Panijpan, Bhinyo; Ruenwongsa, Pintip

    2008-01-01

    An experiment on the action of the yeast, "Saccharomyces cerevisiae", against a fungal plant disease is proposed for secondary students (Grade 11) to support their study of organism interrelationship. This biocontrol experiment serves as the basis for discussing relationships among three organisms (red chilli fruit, "Saccharomyces cerevisiae," and…

  16. Transcriptome analysis of grapefruit flavedo in response to application of the yeast biocontrol agent Metschnikowia fructicola

    USDA-ARS?s Scientific Manuscript database

    Yeasts used to control postharvest pathogens have been shown to induce several biochemical defense responses in surface wounds of fruit. The capability to elicit these responses in fruit tissue has been regarded as one of the possible mechanisms of action by which yeast biocontrol agent inhibit the...

  17. Cucumber rhizosphere microbial community response to biocontrol agent Bacillus subtilis B068150

    USDA-ARS?s Scientific Manuscript database

    Gram-positive bacteria Bacillus subtilis B068150 has been used as a biocontrol agent against the pathogen Fusarium oxysporum f. sp. Cucumerinum. However, their survival ability in cucumber rhizosphere and non-rhizosphere as well as their influence on native microbial communities has not been fully i...

  18. Evaluation of Poultry Litter for Biocontrol of Sclerotia of Sclerotium rolfsii in Soil

    USDA-ARS?s Scientific Manuscript database

    Poultry litter (PL), a major byproduct produced in large quantities on corporate poultry farms for which new uses are needed, was evaluated for potential use as a biocontrol material against sclerotia of Sclerotium rolfsii in soil. Survival of sclerotia was evaluated following their incubation with...

  19. Ecological Considerations in Producing and Formulating Fungal Entomopathogens for Use in Insect Biocontrol

    USDA-ARS?s Scientific Manuscript database

    Insect pests persist in a wide variety of agricultural, arboreal, and urban environments. Effective control with fungal entomopathogens using inundation biocontrol requires an understanding of the ecology of the target insect, fungal pathogen, and the insect-pathogen interaction. Historically, the...

  20. Ecological Considerations in Producing and Formulating Fungal Entomopathogens for Use in Insect Biocontrol

    USDA-ARS?s Scientific Manuscript database

    Insect pests persist in a wide-variety of agricultural, arboreal, and urban environments. Effective control with fungal entomopathogens using inundation biocontrol requires an understanding of the ecology of the target insect, fungal pathogen, and the insect-pathogen interaction. Historically, the d...

  1. Lipopeptides, a novel protein, and volatile compounds contribute to the antifungal activity of the biocontrol agent Bacillus atrophaeus CAB-1.

    PubMed

    Zhang, Xiaoyun; Li, Baoqing; Wang, Ye; Guo, Qinggang; Lu, Xiuyun; Li, Shezeng; Ma, Ping

    2013-11-01

    Bacillus atrophaeus CAB-1 displays a high inhibitory activity against various fungal pathogens and suppresses cucumber powdery mildew and tomato gray mold. We extracted and identified lipopeptides and secreted proteins and volatile compounds produced by strain CAB-1 to investigate the mechanisms involved in its biocontrol performance. In vitro assays indicated all three types of products contributed to the antagonistic activity against the fungal pathogen Botrytis cinerea. Each of these components also effectively prevented the occurrence of the cucumber powdery mildew caused by Sphaerotheca fuliginea under greenhouse conditions. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry revealed that the major bioactive lipopeptide was fengycin A (C15-C17). We isolated the crude-secreted proteins of CAB-1 and purified a fraction with antifungal activity. This protein sequence shared a high identity with a putative phage-related pre-neck appendage protein, which has not been reported as an antifungal factor. The volatile compounds produced by CAB-1 were complex, including a range of alcohols, phenols, amines, and alkane amides. O-anisaldehyde represented one of the most abundant volatiles with the highest inhibition on the mycelial growth of B. cinerea. To our knowledge, this is the first report on profiling three types of antifungal substances in Bacilli and demonstrating their contributions to plant disease control.

  2. Biocontrol of Late Blight (Phytophthora capsici) Disease and Growth Promotion of Pepper by Burkholderia cepacia MPC-7

    PubMed Central

    Sopheareth, Mao; Chan, Sarun; Naing, Kyaw Wai; Lee, Yong Seong; Hyun, Hae Nam; Kim, Young Cheol; Kim, Kil Yong

    2013-01-01

    A chitinolytic bacterial strain having strong antifungal activity was isolated and identified as Burkholderia cepacia MPC-7 based on 16S rRNA gene analysis. MPC-7 solubilized insoluble phosphorous in hydroxyapatite agar media. It produced gluconic acid and 2-ketogluconic acid related to the decrease in pH of broth culture. The antagonist produced benzoic acid (BA) and phenylacetic acid (PA). The authentic compounds, BA and PA, showed a broad spectrum of antimicrobial activity against yeast, several bacterial and fungal pathogens in vitro. To demonstrate the biocontrol efficiency of MPC-7 on late blight disease caused by Phytophthora capsici, pepper plants in pot trials were treated with modified medium only (M), M plus zoospore inoculation (MP), MPC-7 cultured broth (B) and B plus zoospore inoculation (BP). With the sudden increase in root mortality, plants in MP wilted as early as five days after pathogen inoculation. However, plant in BP did not show any symptom of wilting until five days. Root mortality in BP was markedly reduced for as much as 50%. Plants in B had higher dry weight, P concentration in root, and larger leaf area compared to those in M and MP. These results suggested that B. cepacia MPC-7 should be considered as a candidate for the biological fertilizer as well as antimicrobial agent for pepper plants. PMID:25288930

  3. Biocontrol potential of Trichoderma harzianum isolate T-aloe against Sclerotinia sclerotiorum in soybean.

    PubMed

    Zhang, Fuli; Ge, Honglian; Zhang, Fan; Guo, Ning; Wang, Yucheng; Chen, Long; Ji, Xiue; Li, Chengwei

    2016-03-01

    Sclerotinia stem rot, caused by Sclerotinia sclerotiorum (Lib.) de Bary is a major disease of soybean (Glycine max (L.) Merr.). At present, we revealed the three-way interaction between Trichoderma harzianum T-aloe, pathogen S. sclerotiorum and soybean plants in order to demonstrate biocontrol mechanism and evaluate biocontrol potential of T-aloe against S. sclerotiorum in soybean. In our experiments, T-aloe inhibited the growth of S. sclerotiorum with an efficiency of 56.3% in dual culture tests. T-aloe hyphae grew in parallel or intertwined with S. sclerotiorum hyphae and produced hooked contact branches, indicating mycoparasitism. Plate tests showed that T-aloe culture filtrate inhibited S. sclerotiorum growth with an inhibition efficiency of 51.2% and sclerotia production. T-aloe pretreatment showed growth-promoting effect on soybean plants. The activities of peroxidase, superoxide dismutase, and catalase increased, and the hydrogen peroxide (H2O2) as well as the superoxide radical (O2(-)) content in soybean leaves decreased after T-aloe pretreatment in response to S. sclerotiorum pathogen challenge. T-aloe treatment diminished damage caused by pathogen stress on soybean leaf cell membrane, and increased chlorophyll as well as total phenol contents. The defense-related genes PR1, PR2, and PR3 were expressed in the leaves of T-aloe-treated plants. In summary, T-aloe displayed biocontrol potential against S. sclerotiorum. This is the first report of unraveling biocontrol potential of Trichoderma Spp. to soybean sclerotinia stem rot from the three-way interaction between the biocontrol agent, pathogen S. sclerotiorum and soybean plants. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  4. Selection of the N-Acylhomoserine Lactone-Degrading Bacterium Alteromonas stellipolaris PQQ-42 and of Its Potential for Biocontrol in Aquaculture

    PubMed Central

    Torres, Marta; Rubio-Portillo, Esther; Antón, Josefa; Ramos-Esplá, Alfonso A.; Quesada, Emilia; Llamas, Inmaculada

    2016-01-01

    The production of virulence factors by many pathogenic microorganisms depends on the intercellular communication system called quorum sensing, which involves the production and release of signal molecules known as autoinducers. Based on this, new-therapeutic strategies have emerged for the treatment of a variety of infections, such as the enzymatic degradation of signaling molecules, known as quorum quenching (QQ). In this study, we present the screening of QQ activity amongst 450 strains isolated from a bivalve hatchery in Granada (Spain), and the selection of the strain PQQ-42, which degrades a wide range of N-acylhomoserine lactones (AHLs). The selected strain, identified as Alteromonas stellipolaris, degraded the accumulation of AHLs and reduced the production of protease and chitinase and swimming motility of a Vibrio species in co-cultivation experiments in vitro. In the bio-control experiment, strain PQQ-42 significantly reduced the pathogenicity of Vibrio mediterranei VibC-Oc-097 upon the coral Oculina patagonica showing a lower degree of tissue damage (29.25 ± 14.63%) in its presence, compared to when the coral was infected with V. mediterranei VibC-Oc-097 alone (77.53 ± 13.22%). Our results suggest that this AHL-degrading bacterium may have biotechnological applications in aquaculture. PMID:27242684

  5. Selection of the N-Acylhomoserine Lactone-Degrading Bacterium Alteromonas stellipolaris PQQ-42 and of Its Potential for Biocontrol in Aquaculture.

    PubMed

    Torres, Marta; Rubio-Portillo, Esther; Antón, Josefa; Ramos-Esplá, Alfonso A; Quesada, Emilia; Llamas, Inmaculada

    2016-01-01

    The production of virulence factors by many pathogenic microorganisms depends on the intercellular communication system called quorum sensing, which involves the production and release of signal molecules known as autoinducers. Based on this, new-therapeutic strategies have emerged for the treatment of a variety of infections, such as the enzymatic degradation of signaling molecules, known as quorum quenching (QQ). In this study, we present the screening of QQ activity amongst 450 strains isolated from a bivalve hatchery in Granada (Spain), and the selection of the strain PQQ-42, which degrades a wide range of N-acylhomoserine lactones (AHLs). The selected strain, identified as Alteromonas stellipolaris, degraded the accumulation of AHLs and reduced the production of protease and chitinase and swimming motility of a Vibrio species in co-cultivation experiments in vitro. In the bio-control experiment, strain PQQ-42 significantly reduced the pathogenicity of Vibrio mediterranei VibC-Oc-097 upon the coral Oculina patagonica showing a lower degree of tissue damage (29.25 ± 14.63%) in its presence, compared to when the coral was infected with V. mediterranei VibC-Oc-097 alone (77.53 ± 13.22%). Our results suggest that this AHL-degrading bacterium may have biotechnological applications in aquaculture.

  6. Complete Genome Sequence of Micromonospora Strain L5, a Potential Plant-Growth-Regulating Actinomycete, Originally Isolated from Casuarina equisetifolia Root Nodules

    SciTech Connect

    Hirsch, A. M.; Alvarado, J.; Bruce, D.; Chertkov, O.; De Hoff, P. L.; Detter, J. C.; Fujishige, N. A.; Goodwin, L. A.; Han, J.; Han, S.; Ivanova, N.; Land, M. L.; Lum, M. R.; Milani-Nejad, N.; Nolan, M.; Pati, A.; Pitluck, S.; Tran, S. S.; Woyke, T.; Valdes, M.

    2013-08-29

    Micromonospora species live in diverse environments and exhibit a broad range of functions including antibiotic production, biocontrol, and ability to degrade complex polysaccharides. To learn more about these versatile actinomycetes, we sequenced the genome of strain L5, originally isolated from root nodules of an actinorhizal plant growing in Mexico.

  7. Complete Genome Sequence of Micromonospora Strain L5, a Potential Plant-Growth-Regulating Actinomycete, Originally Isolated from Casuarina equisetifolia Root Nodules

    PubMed Central

    Alvarado, Johana; Bruce, David; Chertkov, Olga; De Hoff, Peter L.; Detter, John C.; Fujishige, Nancy A.; Goodwin, Lynne A.; Han, James; Han, Shunsheng; Ivanova, Natalia; Land, Miriam L.; Lum, Michelle R.; Milani-Nejad, Nima; Nolan, Matt; Pati, Amrita; Pitluck, Sam; Tran, Stephen S.; Woyke, Tanja; Valdés, Maria

    2013-01-01

    Micromonospora species live in diverse environments and exhibit a broad range of functions, including antibiotic production, biocontrol, and degradation of complex polysaccharides. To learn more about these versatile actinomycetes, we sequenced the genome of strain L5, originally isolated from root nodules of an actinorhizal plant growing in Mexico. PMID:24072863

  8. Draft Genome Sequence of Streptomyces sp. Strain Wb2n-11, a Desert Isolate with Broad-Spectrum Antagonism against Soilborne Phytopathogens

    DOE PAGES

    Köberl, Martina; White, Richard A.; Erschen, Sabine; ...

    2015-08-06

    Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria, and nematodes. The 8.2-Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants.

  9. Complete genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium of Calendula officinalis

    SciTech Connect

    Köberl, Martina; White, Richard A.; Erschen, Sabine; Spanberger, Nora; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-13

    The genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium (PGPR) with broad-spectrum antagonistic activity against plant-pathogenic fungi, bacteria, and nematodes, consists of a single 3.9-Mb circular chromosome. The genome reveals genes putatively responsible for its promising biocontrol and PGP properties.

  10. Co-inoculating of aflatoxigenic and non-aflatoxigenic strains of Aspergillus flavus to study fungal invasion, colonization, and competition in maize kernels

    USDA-ARS?s Scientific Manuscript database

    A currently utilized pre-harvest bio-control method involves field inoculations with non-aflatoxigenic Aspergillus flavus, a tactic shown to strategically displace the native aflatoxin producing strain and effectively decrease aflatoxin contamination in corn. The present in situ study focuses on tra...

  11. Draft Genome Sequence of Streptomyces sp. Strain Wb2n-11, a Desert Isolate with Broad-Spectrum Antagonism against Soilborne Phytopathogens

    PubMed Central

    White, Richard A.; Erschen, Sabine; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-01-01

    Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria, and nematodes. The 8.2-Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants. PMID:26251492

  12. Draft Genome Sequence of Streptomyces sp. Strain Wb2n-11, a Desert Isolate with Broad-Spectrum Antagonism against Soilborne Phytopathogens

    SciTech Connect

    Köberl, Martina; White, Richard A.; Erschen, Sabine; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-06

    Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria, and nematodes. The 8.2-Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants.

  13. Draft Genome Sequence of Streptomyces sp. Strain Wb2n-11, a Desert Isolate with Broad-Spectrum Antagonism against Soilborne Phytopathogens

    SciTech Connect

    Köberl, Martina; White, Richard A.; Erschen, Sabine; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-06

    Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria and nematodes. The 8.2 Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants.

  14. The genome sequence of the biocontrol fungus Metarhizium anisopliae and comparative genomics of Metarhizium species.

    PubMed

    Pattemore, Julie A; Hane, James K; Williams, Angela H; Wilson, Bree A L; Stodart, Ben J; Ash, Gavin J

    2014-08-07

    Metarhizium anisopliae is an important fungal biocontrol agent of insect pests of agricultural crops. Genomics can aid the successful commercialization of biopesticides by identification of key genes differentiating closely related species, selection of virulent microbial isolates which are amenable to industrial scale production and formulation and through the reduction of phenotypic variability. The genome of Metarhizium isolate ARSEF23 was recently published as a model for M. anisopliae, however phylogenetic analysis has since re-classified this isolate as M. robertsii. We present a new annotated genome sequence of M. anisopliae (isolate Ma69) and whole genome comparison to M. robertsii (ARSEF23) and M. acridum (CQMa 102). Whole genome analysis of M. anisopliae indicates significant macrosynteny with M. robertsii but with some large genomic inversions. In comparison to M. acridum, the genome of M. anisopliae shares lower sequence homology. While alignments overall are co-linear, the genome of M. acridum is not contiguous enough to conclusively observe macrosynteny. Mating type gene analysis revealed both MAT1-1 and MAT1-2 genes present in M. anisopliae suggesting putative homothallism, despite having no known teleomorph, in contrast with the putatively heterothallic M. acridum isolate CQMa 102 (MAT1-2) and M. robertsii isolate ARSEF23 (altered MAT1-1). Repetitive DNA and RIP analysis revealed M. acridum to have twice the repetitive content of the other two species and M. anisopliae to be five times more RIP affected than M. robertsii. We also present an initial bioinformatic survey of candidate pathogenicity genes in M. anisopliae. The annotated genome of M. anisopliae is an important resource for the identification of virulence genes specific to M. anisopliae and development of species- and strain- specific assays. New insight into the possibility of homothallism and RIP affectedness has important implications for the development of M. anisopliae as a

  15. Growth kinetics and efficacy as parameters for ranking and selecting biocontrol agents that reduce pink rot in stored potatoes

    USDA-ARS?s Scientific Manuscript database

    Increased production of organic agricultural products and the relative ineffectiveness of traditional control measures support development of new biocontrol technologies for use against pink rot infections in storage. The microbiota of 84 different agricultural soils was individually transferred to...

  16. Lentinula edodes enhances the biocontrol activity of Cryptococcus laurentii against Penicillium expansum contamination and patulin production in apple fruits.

    PubMed

    Tolaini, V; Zjalic, S; Reverberi, M; Fanelli, C; Fabbri, A A; Del Fiore, A; De Rossi, P; Ricelli, A

    2010-04-15

    Penicillium expansum is a post-harvest pathogen of apples which can produce the hazardous mycotoxin patulin. The yeast Cryptococcus laurentii (LS28) is a biocontrol agent able to colonize highly oxidative environments such as wounds in apples. In this study culture filtrates of the basidiomycete Lentinula edodes (LF23) were used to enhance the biocontrol activity of LS28. In vitro L. edodes culture filtrates improved the growth of C. laurentii and the activity of its catalase, superoxide dismutase and glutathione peroxidase, which play a key role in oxidant scavenging. In addition, LF23 also delayed P. expansum conidia germination. The biocontrol effect of LS28 used together with LF23 in wounded apples improved the inhibition of P. expansum growth and patulin production in comparison with LS28 alone, under both experimental and semi-commercial conditions. The biocontrol effect was confirmed by a semi-quantitative PCR analysis set up for monitoring the growth of P. expansum.

  17. Phenotypic comparison of clinical and plant-beneficial strains of Pantoea agglomerans.

    PubMed

    Bonaterra, Anna; Badosa, Esther; Rezzonico, Fabio; Duffy, Brion; Montesinos, Emilio

    2014-06-01

    Certain strains of Pantoea are used as biocontrol agents for the suppression of plant diseases. However, their commercial registration is hampered in some countries because of biosafety concerns. This study compares clinical and plant-beneficial strains of P. agglomerans and related species using a phenotypic analysis approach in which plant-beneficial effects, adverse effects in nematode models, and toxicity were evaluated. Plant-beneficial effects were determined as the inhibition of apple fruit infection by Penicillium expansum and apple flower infection by Erwinia amylovora. Clinical strains had no general inhibitory activity against infection by the fungal or bacterial plant pathogens, as only one clinical strain inhibited P. expansum and three inhibited E. amylovora. By contrast, all biocontrol strains showed activity against at least one of the phytopathogens, and three strains were active against both. The adverse effects in animals were evaluated in the plant-parasitic nematode Meloidogyne javanica and the bacterial-feeding nematode Caenorhabditis elegans. Both models indicated adverse effects of the two clinical strains but not of any of the plant-beneficial strains. Toxicity was evaluated by means of hemolytic activity in blood, and genotoxicity with the Ames test. None of the strains, whether clinical or plant-beneficial, showed any evidence of toxicity. Copyright© by the Spanish Society for Microbiology and Institute for Catalan Studies.

  18. Colonization of Tomato Plants by Two Agrocin-Producing Strains of Agrobacterium tumefaciens

    PubMed Central

    Macrae, Sharmane; Thomson, Jennifer A.; Van Staden, Johannes

    1988-01-01

    For a bacterium to be a successful biocontrol agent against crown gall disease, it must produce an effective agrocin specific for Agrobacterium tumefaciens and be able to colonize host plants efficiently. The colonization abilities of K84 and J73, successful and potential biocontrolling strains, respectively, were compared both in vivo and in vitro. Both strains produced fibrils attaching them to tomato root surfaces and had similar colonization efficiencies up to 14 days after inoculation. However, the ability of J73 to colonize plants for longer periods was significantly less than that of K84. Thus, the presence of fibrils is not sufficient to ensure colonization. No correlation was found between hydrophobicity and colonization. Images PMID:16347802

  19. The Potential Application and Risks Associated With the Use of Predatory Bacteria as a Biocontrol Agent Against Wound Infections

    DTIC Science & Technology

    2014-09-01

    Associated With the Use of Predatory Bacteria as a Biocontrol Agent Against Wound Infections PRINCIPAL INVESTIGATOR: Daniel E Kadouri, Ph.D...W81XWH-12-2-0067 4. TITLE AND SUBTITLE The Potential Application and Risks Associated With the Use of Predatory Bacteria as a Biocontrol Agent...serve as a novel therapeutic agent to control wound-related bacterial infections. In a previous study, we confirmed that predatory bacteria Bdellovibrio

  20. Isolation and Characterization of phiLLS, a Novel Phage with Potential Biocontrol Agent against Multidrug-Resistant Escherichia coli

    PubMed Central

    Amarillas, Luis; Rubí-Rangel, Lucia; Chaidez, Cristobal; González-Robles, Arturo; Lightbourn-Rojas, Luis; León-Félix, Josefina

    2017-01-01

    Foodborne diseases are a serious and growing problem, and the incidence and prevalence of antimicrobial resistance among foodborne pathogens is reported to have increased. The emergence of antibiotic-resistant bacterial strains demands novel strategies to counteract this epidemic. In this regard, lytic bacteriophages have reemerged as an alternative for the control of pathogenic bacteria. However, the effective use of phages relies on appropriate biological and genomic characterization. In this study, we present the isolation and characterization of a novel bacteriophage named phiLLS, which has shown strong lytic activity against generic and multidrug-resistant Escherichia coli strains. Transmission electron microscopy of phiLLS morphology revealed that it belongs to the Siphoviridae family. Furthermore, this phage exhibited a relatively large burst size of 176 plaque-forming units per infected cell. Phage phiLLS significantly reduced the growth of E. coli under laboratory conditions. Analyses of restriction profiles showed the presence of submolar fragments, confirming that phiLLS is a pac-type phage. Phylogenetic analysis based on the amino acid sequence of large terminase subunits confirmed that this phage uses a headful packaging strategy to package their genome. Genomic sequencing and bioinformatic analysis showed that phiLLS is a novel bacteriophage that is most closely related to T5-like phages. In silico analysis indicated that the phiLLS genome consists of 107,263 bp (39.0 % GC content) encoding 160 putative ORFs, 16 tRNAs, several potential promoters and transcriptional terminators. Genome analysis suggests that the phage phiLLS is strictly lytic without carrying genes associated with virulence factors and/or potential immunoreactive allergen proteins. The bacteriophage isolated in this study has shown promising results in the biocontrol of bacterial growth under in vitro conditions, suggesting that it may prove useful as an alternative agent for the

  1. Draft Genome Sequence of Plant Growth-Promoting Rhizobacterium Pantoea sp. Strain AS-PWVM4.

    PubMed

    Khatri, Indu; Kaur, Sukhvir; Devi, Usha; Kumar, Navinder; Sharma, Deepak; Subramanian, Srikrishna; Saini, Adesh K

    2013-12-05

    Nonpathogenic Pantoea spp. have been shown to confer biofertilizer and biocontrol activities, indicating their potential for increasing crop yield. Herein, we provide the high-quality genome sequence of Pantoea sp. strain AS-PWVM4, a Gram-negative motile plant growth-promoting rhizobacterium isolated from a pomegranate plant. The 4.9-Mb genome contains genes related to plant growth promotion and the synthesis of siderophores.

  2. Biocontrol Potentials of Antimicrobial Peptide Producing Bacillus Species: Multifaceted Antagonists for the Management of Stem Rot of Carnation Caused by Sclerotinia sclerotiorum

    PubMed Central

    Vinodkumar, S.; Nakkeeran, S.; Renukadevi, P.; Malathi, V. G.

    2017-01-01

    Bacillus species are widely exploited as biocontrol agents because of their efficiency in impeding various plant pathogens with multifaceted approach. In this study, Bacillus species were isolated from rhizosphere of various plants viz., carnations, cotton, turmeric, and bananas in Tamil Nadu state of India. Their potential to control the mycelial growth of Sclerotinia sclerotiorum was assessed in vitro by dual plate and partition plate techniques. B. amyloliquefaciens strain VB7 was much effective in inhibiting mycelial growth (45% inhibition of over control) and sclerotial production (100%). PCR detection of AMP genes revealed that B. amyloliquefaciens (VB7) had a maximum of 10 diverse antibiotic biosynthesis genes, namely, ituD, ipa14, bacA, bacD, bamC, sfP, spaC, spaS, alba, and albF, that resulted in production of the antibiotics iturin, bacilysin, bacillomycin, surfactin, subtilin, and subtilosin. Further, metabolites from B. amyloliquefaciens strains VB2 and VB7, associated with inhibition of S. sclerotiorum, were identified as phenols and fatty acids by gas chromatography mass spectrometry (GC-MS). Delivery of bacterial suspension of the effective strains of Bacillus spp. as root dip was found promising for the management of stem rot of cultivated carnations. Minimal percent disease incidence (4.6%) and maximum plant growth promotion was observed in the plants treated with B. amyloliquefaciens (VB7). PMID:28392780

  3. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7

    PubMed Central

    Maldonado-González, M. Mercedes; Bakker, Peter A. H. M.; Prieto, Pilar; Mercado-Blanco, Jesús

    2015-01-01

    The effective management of Verticillium wilts (VW), diseases affecting many crops and caused by some species of the soil-borne fungus Verticillium, is problematic. The use of microbial antagonists to control these pathologies fits modern sustainable agriculture criteria. Pseudomonas fluorescens PICF7 is an endophytic bacterium isolated from olive roots with demonstrated ability to control VW of olive caused by the highly virulent, defoliating (D) pathotype of Verticillium dahliae Kleb. However, the study of the PICF7-V. dahliae-olive tripartite interaction poses difficulties because of the inherent characteristics of woody, long-living plants. To overcome these problems we explored the use of the model plant Arabidopsis thaliana. Results obtained in this study showed that: (i) olive D and non-defoliating V. dahliae pathotypes produce differential disease severity in A. thaliana plants; (ii) strain PICF7 is able to colonize and persist in the A. thaliana rhizosphere but is not endophytic in Arabidopsis; and (iii) strain PICF7 controls VW in Arabidopsis. Additionally, as previously observed in olive, neither swimming motility nor siderophore production by PICF7 are required for VW control in A. thaliana, whilst cysteine auxotrophy decreased the effectiveness of PICF7. Moreover, when applied to the roots PICF7 controlled Botrytis cinerea infection in the leaves of Arabidopsis, suggesting that this strain is able to induce systemic resistance. A. thaliana is therefore a suitable alternative to olive bioassays to unravel biocontrol traits involved in biological control of V. dahliae by P. fluorescens PICF7. PMID:25904904

  4. Population dynamics of Aspergillus flavus following biocontrol treatment of corn

    USDA-ARS?s Scientific Manuscript database

    Aspergillus flavus is a fungal pathogen of many agronomically important crops worldwide. We sampled A. flavus strains from a cornfield in Rocky Mount, North Carolina, over a period of two years. The field was planted in 2010 and plots were inoculated at tasselling with either AF36 or NRRL 21882 (=Af...

  5. Disruption of Gene pqqA or pqqB Reduces Plant Growth Promotion Activity and Biocontrol of Crown Gall Disease by Rahnella aquatilis HX2

    PubMed Central

    Hale, Lauren; Wu, Wenliang; Guo, Yanbin

    2014-01-01

    Rahnella aquatilis strain HX2 has the ability to promote maize growth and suppress sunflower crown gall disease caused by Agrobacterium vitis, A. tumefaciens, and A. rhizogenes. Pyrroloquinoline quinone (PQQ), a cofactor of aldose and alcohol dehydrogenases, is required for the synthesis of an antibacterial substance, gluconic acid, by HX2. Mutants of HX2 unable to produce PQQ were obtained by in-frame deletion of either the pqqA or pqqB gene. In this study, we report the independent functions of pqqA and pqqB genes in relation to PQQ synthesis. Interestingly, both the pqqA and pqqB mutants of R. aquatilis eliminated the ability of strain HX2 to produce antibacterial substance, which in turn, reduced the effectiveness of the strain for biological control of sunflower crown gall disease. The mutation also resulted in decreased mineral phosphate solubilization by HX2, which reduced the efficacy of this strain as a biological fertilizer. These functions were restored by complementation with the wild-type pqq gene cluster. Additionally, the phenotypes of HX2 derivatives, including colony morphology, growth dynamic, and pH change of culture medium were impacted to different extents. Our findings suggested that pqqA and pqqB genes individually play important functions in PQQ biosynthesis and are required for antibacterial activity and phosphorous solubilization. These traits are essential for R. aquatilis efficacy as a biological control and plant growth promoting strain. This study enhances our fundamental understanding of the biosynthesis of an environmentally significant cofactor produced by a promising biocontrol and biological fertilizer strain. PMID:25502691

  6. Comparative Genomics of Bacillus amyloliquefaciens Strains Reveals a Core Genome with Traits for Habitat Adaptation and a Secondary Metabolites Rich Accessory Genome

    PubMed Central

    Belbahri, Lassaad; Chenari Bouket, Ali; Rekik, Imen; Alenezi, Faizah N.; Vallat, Armelle; Luptakova, Lenka; Petrovova, Eva; Oszako, Tomasz; Cherrad, Semcheddine; Vacher, Sébastien; Rateb, Mostafa E.

    2017-01-01

    The Gram positive, non-pathogenic endospore-forming soil inhabiting prokaryote Bacillus amyloliquefaciens is a plant growth-promoting rhizobacterium. Bacillus amyloliquefaciens processes wide biocontrol abilities and numerous strains have been reported to suppress diverse bacterial, fungal and fungal-like pathogens. Knowledge about strain level biocontrol abilities is warranted to translate this knowledge into developing more efficient biocontrol agents and bio-fertilizers. Ever-expanding genome studies of B. amyloliquefaciens are showing tremendous increase in strain-specific new secondary metabolite clusters which play key roles in the suppression of pathogens and plant growth promotion. In this report, we have used genome mining of all sequenced B. amyloliquefaciens genomes to highlight species boundaries, the diverse strategies used by different strains to promote plant growth and the diversity of their secondary metabolites. Genome composition of the targeted strains suggest regions of genomic plasticity that shape the structure and function of these genomes and govern strain adaptation to different niches. Our results indicated that B. amyloliquefaciens: (i) suffer taxonomic imprecision that blurs the debate over inter-strain genome diversity and dynamics, (ii) have diverse strategies to promote plant growth and development, (iii) have an unlocked, yet to be delimited impressive arsenal of secondary metabolites and products, (iv) have large number of so-called orphan gene clusters, i.e., biosynthetic clusters for which the corresponding metabolites are yet unknown, and (v) have a dynamic pan genome with a secondary metabolite rich accessory genome. PMID:28824571

  7. The requirement for the LysR-type regulator PtrA for Pseudomonas chlororaphis PA23 biocontrol revealed through proteomic and phenotypic analysis

    PubMed Central

    2014-01-01

    Background Pseudomonas chlororaphis strain PA23 is a biocontrol agent capable of suppressing the fungal pathogen Sclerotinia sclerotiorum. This bacterium produces the antibiotics phenazine and pyrrolnitrin together with other metabolites believed to contribute to biocontrol. A mutant no longer capable of inhibiting fungal growth was identified harboring a transposon insertion in a gene encoding a LysR-type transcriptional regulator (LTTR), designated ptrA (Pseudomonas transcriptional regulator). Isobaric tag for relative and absolute quantitation (iTRAQ) based protein analysis was used to reveal changes in protein expression patterns in the ptrA mutant compared to the PA23 wild type. Results Relative abundance profiles showed 59 differentially-expressed proteins in the ptrA mutant, which could be classified into 16 clusters of orthologous groups (COGs) based on their predicted functions. The largest COG category was the unknown function group, suggesting that many yet-to-be identified proteins are involved in the loss of fungal activity. In the secondary metabolite biosynthesis, transport and catabolism COG, seven proteins associated with phenazine biosynthesis and chitinase production were downregulated in the mutant. Phenotypic assays confirmed the loss of phenazines and chitinase activity. Upregulated proteins included a lipoprotein involved in iron transport, a flagellin and hook-associated protein and four proteins categorized into the translation, ribosome structure and biogenesis COG. Phenotypic analysis revealed that the mutant exhibited increased siderophore production and flagellar motility and an altered growth profile, supporting the proteomic findings. Conclusion PtrA is a novel LTTR that is essential for PA23 fungal antagonism. Differential protein expression was observed across 16 COG categories suggesting PtrA is functioning as a global transcriptional regulator. Changes in protein expression were confirmed by phenotypic assays that showed reduced

  8. Use of green fluorescent protein-based reporters to monitor balanced production of antifungal compounds in the biocontrol agent Pseudomonas fluorescens CHA0.

    PubMed

    Baehler, E; Bottiglieri, M; Péchy-Tarr, M; Maurhofer, M; Keel, C

    2005-01-01

    To develop reporter constructs based on stable and unstable variants of the green fluorescent protein (GFP) for monitoring balanced production of antifungal compounds that are crucial for the capacity of the root-colonizing Pseudomonas fluorescens strain CHA0 to control plant diseases caused by soil-borne pathogenic fungi. Pseudomonas fluorescens CHA0 produces the three antifungal metabolites 2,4-diacetylphloroglucinol (DAPG), pyoluteorin (PLT) and pyrrolnitrin (PRN). The gfp[mut3] and gfp[AAV] reporter genes were fused to the promoter regions of the DAPG, PLT and PRN biosynthetic genes. The reporter fusions were then used to follow the kinetics of expression of the three antifungal metabolites in a microplate assay. DAPG and PLT were found to display an inverse relationship in which each metabolite activates its own biosynthesis while repressing the synthesis of the other metabolite. PRN appears not to be involved in this balance. However, the microbial and plant phenolic metabolite salicylate was found to interfere with the expression of both DAPG and PLT. The results obtained provide evidence that P. fluorescens CHA0 may keep the antifungal compounds DAPG and PLT at a fine-tuned balance that can be affected by certain microbial and plant phenolics. To our knowledge, the present study is the first to use stable and unstable GFP variants to study antibiotic gene expression in a biocontrol pseudomonad. The developed reporter fusions will be a highly valuable tool to study in situ expression of this bacterial biocontrol trait on plant roots, i.e. at the site of pathogen suppression.

  9. Genetic engineering of fungal biocontrol agents to achieve greater efficacy against insect pests.

    PubMed

    St Leger, Raymond J; Wang, Chengshu

    2010-01-01

    Molecular biology methods have elucidated pathogenic processes in several fungal biocontrol agents including two of the most commonly applied entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana. In this review, we describe how a combination of molecular techniques has: (1) identified and characterized genes involved in infection; (2) manipulated the genes of the pathogen to improve biocontrol performance; and (3) allowed expression of a neurotoxin from the scorpion Androctonus australis. The complete sequencing of four exemplar species of entomopathogenic fungi including B. bassiana and M. anisopliae will be completed in 2010. Coverage of these genomes will help determine the identity, origin, and evolution of traits needed for diverse lifestyles and host switching. Such knowledge combined with the precision and malleability of molecular techniques will allow design of multiple pathogens with different strategies to be used for different ecosystems and avoid the possibility of the host developing resistance.

  10. A transformation system for the biocontrol yeast, Candida oleophila, based on hygromycin B resistance.

    PubMed

    Yehuda, H; Droby, S; Wisniewski, M; Goldway, M

    2001-12-01

    Lithium acetate transformation and electroporation were applied to the biocontrol yeast, Candida oleophila. The hygromycin B resistance gene, flanked by the phosphoglycerate kinase promoter and terminator of Candida tropicalis, served as the genetic selection marker. The transformation efficiency of electroporation was almost 400 times more efficient than that of the lithium acetate method. While incorporation of DNA, flanked by a sequence endogenous to C. oleophila, transpired apparently by homologous recombination, the integration of DNA (that did not contain C. oleophila DNA) occurred at random. Whereas transformants were observed with a linear segment of the plasmid, none were detected with the undigested plasmid. This system provides both a tool for the molecular analysis of the biocontrol mechanism of C. oleophila and a means of tagging C. oleophila for field studies.

  11. Biocontrol efficacy of different isolates of Trichoderma against soil borne pathogen Rhizoctonia solani.

    PubMed

    Asad, Saeed Ahmad; Ali, Naeem; Hameed, Abdul; Khan, Sabaz Ali; Ahmad, Rafiq; Bilal, Muhammad; Shahzad, Muhammad; Tabassum, Ayesha

    2014-01-01

    In this study, the biocontrol abilities of water-soluble and volatile metabolites of three different isolates of Trichoderma (T. asperellum, T. harzianum and Trichoderma spp.) against soil borne plant pathogen Rhizoctonia solani were investigated both in vitro and in vivo. The results showed for the first time that mycelial growth inhibition of the pathogen was 74.4-67.8% with water-soluble metabolites as compared to 15.3-10.6% with volatile metabolites in vitro. In vivo antagonistic activity of Trichoderma isolates against R. solani was evaluated on bean plants under laboratory and greenhouse conditions. We observed that T. asperellum was more effective and consistent, lowering disease incidence up to 19.3% in laboratory and 30.5% in green house conditions. These results showed that three isolates of Trichoderma could be used as effective biocontrol agents against R. solani.

  12. Unrecognized impact of a biocontrol agent on the spread rate of an invasive thistle.

    PubMed

    Marchetto, Katherine M; Shea, Katriona; Kelly, Dave; Groenteman, Ronny; Sezen, Zeynep; Jongejans, Eelke

    2014-07-01

    Herbivores may significantly reduce plant populations by reducing seed set; however, we know little of their impact on seed movement. We show for the first time that the receptacle-feeding weevil Rhinocyllus conicus not only reduces seed production by the invasive thistle Carduus nutans but also inhibits release and subsequent wind dispersal of seeds. These effects generate large, though different, impacts on spatial spread and local abundance in two populations with differing demography, located in the United States and New Zealand. Furthermore, the mechanism is context dependent, with the largest effects through increased terminal velocity in the United States but through reduced seed production in New Zealand. Our results show that the benefit of biocontrol programs may have been underestimated; screenings of potential biocontrol agents should examine effects on pest dispersal and spread, as well as on abundance.

  13. Endophytic fungi associated with Monarda citriodora, an aromatic and medicinal plant and their biocontrol potential.

    PubMed

    Katoch, Meenu; Pull, Shipra

    2017-12-01

    The Food and Agriculture Organization has estimated that every year considerable losses of the food crops occur due to plant diseases. Although fungicides are extensively used for management of plant diseases, they are expensive and hazardous to the environment and human health. Alternatively, biological control is the safe way to overcome the effects of plant diseases and to sustain agriculture. Since Monarda citriodora Cerv. ex Lag. (Lamiaceae/Labiatae) is known for its antifungal properties, it was chosen for the study. The isolation of endophytic fungi from M. citriodora and assessing their biocontrol potential. The isolated endophytes were characterized using ITS-5.8 S rDNA sequencing. Their biocontrol potential was assessed using different antagonistic assays against major plant pathogens. Twenty-eight endophytes representing 11 genera were isolated, of which, around 82% endophytes showed biocontrol potential against plant pathogens. MC-2 L (Fusarium oxysporum), MC-14 F (F. oxysporum), MC-22 F (F. oxysporum) and MC-25 F (F. redolens) displayed significant antagonistic activity against all the tested pathogens. Interestingly, MC-10 L (Muscodor yucatanensis) completely inhibited the growth of Sclerotinia sp., Colletotrichum capsici, Aspergillus flavus and A. fumigatus in dual culture assay, whereas MC-8 L (A. oryzae) and MC-9 L (Penicillium commune) completely inhibited the growth of the Sclerotinia sp. in fumigation assay. Endophytes MC-2 L, MC-14 F, MC-22 F and MC-25 F could effectively be used to control broad range of phytopathogens, while MC-10 L, MC-8 L and MC-9 L could be used to control specific pathogens. Secondly, endophytes showing varying degrees of antagonism in different assays represented the chemo-diversity not only as promising biocontrol agents but also as a resource of defensive and bioactive metabolites.

  14. Increasing corn for biofuel production reduces biocontrol services in agricultural landscapes

    PubMed Central

    Landis, Douglas A.; Gardiner, Mary M.; van der Werf, Wopke; Swinton, Scott M.

    2008-01-01

    Increased demand for corn grain as an ethanol feedstock is altering U.S. agricultural landscapes and the ecosystem services they provide. From 2006 to 2007, corn acreage increased 19% nationally, resulting in reduced crop diversity in many areas. Biological control of insects is an ecosystem service that is strongly influenced by local landscape structure. Here, we estimate the value of natural biological control of the soybean aphid, a major pest in agricultural landscapes, and the economic impacts of reduced biocontrol caused by increased corn production in 4 U.S. states (Iowa, Michigan, Minnesota, and Wisconsin). For producers who use an integrated pest management strategy including insecticides as needed, natural suppression of soybean aphid in soybean is worth an average of $33 ha−1. At 2007–2008 prices these services are worth at least $239 million y−1 in these 4 states. Recent biofuel-driven growth in corn planting results in lower landscape diversity, altering the supply of aphid natural enemies to soybean fields and reducing biocontrol services by 24%. This loss of biocontrol services cost soybean producers in these states an estimated $58 million y−1 in reduced yield and increased pesticide use. For producers who rely solely on biological control, the value of lost services is much greater. These findings from a single pest in 1 crop suggest that the value of biocontrol services to the U.S. economy may be underestimated. Furthermore, we suggest that development of cellulosic ethanol production processes that use a variety of feedstocks could foster increased diversity in agricultural landscapes and enhance arthropod-mediated ecosystem services. PMID:19075234

  15. [Ecological aspects of mosquito biocontrol with implementation of GPS/GIS].

    PubMed

    Lonc, Elzbieta; Rydzanicz, Katarzyna; Jawień, Piotr

    2010-01-01

    Mosquitoes bionomics and vector role as well as integrated control methods with GPS/GIS techniques were presented. Special attention was put on GIS which enables analysis of biological and environmental data generated by GPS (Global Positioning System). Combined with data from surveillance and management activities, those techniques provide a powerful tool for the precise analysis of mosquito development, breeding sites, and effective biocontrol effects on maps.

  16. Genetic Transformation of the Biocontrol Fungus Gliocladium virens to Benomyl Resistance

    PubMed Central

    Ossanna, Nina; Mischke, Sue

    1990-01-01

    Methodology was developed to isolate and regenerate protoplasts from the biocontrol fungus Gliocladium virens and to transform them to benomyl resistance with a Neurospora crassa β-tubulin gene. Southern blots demonstrated that multiple copies of the vector integrated into the chromosomal DNA of stable biotypes but not of abortive transformants. Analysis of nuclear condition in vegetative and asexual structures demonstrated that no structure of G. virens is dependably uninucleate and thus preferentially suitable for transformation. Images PMID:16348312

  17. Cannibalism in invasive, native and biocontrol populations of the harlequin ladybird

    PubMed Central

    2014-01-01

    Background Cannibalism is widespread in both vertebrates and invertebrates but its extent is variable between and within species. Cannibalism depends on population density and nutritional conditions, and could be beneficial during colonisation of new environments. Empirical studies are needed to determine whether this trait might facilitate invasion of a new area in natural systems. We investigated whether the propensity for cannibalism in H. axyridis differs both between native and invasive populations and between invasive populations from the core and from the front of the invasive area in Western Europe. We also compared the propensity for cannibalism of these natural populations with that of laboratory-reared biocontrol populations. We measured the cannibalism rates of eggs by first instar larvae and adult females at two different individual densities of ladybirds from three types of population (invasive, native and biocontrol), in laboratory-controlled conditions. Results Cannibalism was significantly greater in larvae from invasive populations compared to native or biocontrol populations, but there was no difference in cannibalism rates between populations from the core or front of the invaded range. Cannibalism was significantly lower in larvae from biocontrol populations compared to wild (invasive and native) populations. No differences in cannibalism rates of adult females were found between any populations. While high population density significantly increased cannibalism in both larvae and adults, the norm of reaction of cannibalism to individual density did not change significantly during the invasion and/or laboratory rearing processes. Conclusion This study is the first to provide evidence for a higher propensity for cannibalism in invasive populations compared to native ones. Our experiments also shed light on the difference in cannibalism evolution with respect to life stages. However, we are still at an early stage in understanding the underlying

  18. Cannibalism in invasive, native and biocontrol populations of the harlequin ladybird.

    PubMed

    Tayeh, Ashraf; Estoup, Arnaud; Lombaert, Eric; Guillemaud, Thomas; Kirichenko, Natalia; Lawson-Handley, Lori; De Clercq, Patrick; Facon, Benoît

    2014-02-05

    Cannibalism is widespread in both vertebrates and invertebrates but its extent is variable between and within species. Cannibalism depends on population density and nutritional conditions, and could be beneficial during colonisation of new environments. Empirical studies are needed to determine whether this trait might facilitate invasion of a new area in natural systems. We investigated whether the propensity for cannibalism in H. axyridis differs both between native and invasive populations and between invasive populations from the core and from the front of the invasive area in Western Europe. We also compared the propensity for cannibalism of these natural populations with that of laboratory-reared biocontrol populations. We measured the cannibalism rates of eggs by first instar larvae and adult females at two different individual densities of ladybirds from three types of population (invasive, native and biocontrol), in laboratory-controlled conditions. Cannibalism was significantly greater in larvae from invasive populations compared to native or biocontrol populations, but there was no difference in cannibalism rates between populations from the core or front of the invaded range. Cannibalism was significantly lower in larvae from biocontrol populations compared to wild (invasive and native) populations. No differences in cannibalism rates of adult females were found between any populations. While high population density significantly increased cannibalism in both larvae and adults, the norm of reaction of cannibalism to individual density did not change significantly during the invasion and/or laboratory rearing processes. This study is the first to provide evidence for a higher propensity for cannibalism in invasive populations compared to native ones. Our experiments also shed light on the difference in cannibalism evolution with respect to life stages. However, we are still at an early stage in understanding the underlying mechanisms and several

  19. Plant microbial diversity is suggested as the key to future biocontrol and health trends.

    PubMed

    Berg, Gabriele; Köberl, Martina; Rybakova, Daria; Müller, Henry; Grosch, Rita; Smalla, Kornelia

    2017-05-01

    The microbiome of plants plays a crucial role in both plant and ecosystem health. Rapid advances in multi-omics tools are dramatically increasing access to the plant microbiome and consequently to the identification of its links with diseases and to the control of those diseases. Recent insights reveal a close, often symbiotic relationship between microorganisms and plants. Microorganisms can stimulate germination and plant growth, prevent diseases, and promote stress resistance and general fitness. Plants and their associated microorganisms form a holobiont and have to be considered as co-evolved species assemblages consisting of bacterial, archaeal and diverse eukaryotic species. The beneficial interplay of the host and its microbiome is responsible for maintaining the health of the holobiont, while diseases are often correlated with microbial dysbioses. Microbial diversity was identified as a key factor in preventing diseases and can be implemented as a biomarker in plant protection strategies. Targeted and predictive biocontrol approaches are possible by developing microbiome-based solutions. Moreover, combined breeding and biocontrol strategies maintaining diversity and ecosystem health are required. The analysis of plant microbiome data has brought about a paradigm shift in our understanding of its role in health and disease and has substantial consequences for biocontrol and health issues. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Influence of Oxidative Stress on Biocontrol Activity of Cryptococcus laurentii against Blue Mold on Peach Fruit

    PubMed Central

    Zhang, Zhanquan; Chen, Jian; Li, Boqiang; He, Chang; Chen, Yong; Tian, Shiping

    2017-01-01

    The limitations of chemical fungicides for the control of postharvest diseases have recently become more apparent. The utilization of antagonistic microorganisms is a promising alternative to that of fungicides to control postharvest decay. In previous studies, the antagonistic yeast Cryptococcus laurentii has shown excellent effects of biocontrol and great potential for practical application. Adverse conditions, such as oxidative stress, limit the practical application of antagonistic yeast. In this study, we investigated the oxidative stress tolerance of C. laurentii and the associated mechanisms. The results indicated that exogenous oxidative stress has a significant effect on the viability and biocontrol efficiency of C. laurentii. H2O2-induced oxidative stress led to the accumulation of reactive oxygen species. The results of flow cytometric analysis suggested that apoptosis is responsible for the reduced survival rate of C. laurentii under oxidative stress. Using tests of antioxidant activity, we found that C. laurentii could employ enzymatic systems to resist exogenous oxidative stress. The addition of exogenous glutathione, a non-enzymatic antioxidant, to the media can significantly enhance oxidative tolerance and biocontrol efficiency of C. laurentii. PMID:28210254

  1. Mechanism of H₂O₂-induced oxidative stress regulating viability and biocontrol ability of Rhodotorula glutinis.

    PubMed

    Chen, Jian; Li, Boqiang; Qin, Guozheng; Tian, Shiping

    2015-01-16

    The use of antagonistic yeasts to control postharvest pathogens is a promising alternative to fungicides. The effectiveness of the antagonists against fungal pathogens is greatly dependent on their viability, which is usually mediated by reactive oxygen species (ROS). Here, we investigated the effects of H₂O₂-induced oxidative stress on the viability and biocontrol efficacy of Rhodotorula glutinis and, using flow cytometric analysis, observed the changes of ROS accumulation and apoptosis in the yeast cells with or without H₂O