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Sample records for phytopathogen verticillium longisporum

  1. Identification and Differentiation of Verticillium Species and V. longisporum Lineages by Simplex and Multiplex PCR Assays

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    Inderbitzin, Patrik; Davis, R. Michael; Bostock, Richard M.; Subbarao, Krishna V.

    2013-01-01

    Accurate species identification is essential for effective plant disease management, but is challenging in fungi including Verticillium sensu stricto (Ascomycota, Sordariomycetes, Plectosphaerellaceae), a small genus of ten species that includes important plant pathogens. Here we present fifteen PCR assays for the identification of all recognized Verticillium species and the three lineages of the diploid hybrid V. longisporum. The assays were based on DNA sequence data from the ribosomal internal transcribed spacer region, and coding and non-coding regions of actin, elongation factor 1-alpha, glyceraldehyde-3-phosphate dehydrogenase and tryptophan synthase genes. The eleven single target (simplex) PCR assays resulted in amplicons of diagnostic size for V. alfalfae, V. albo-atrum, V. dahliae including V. longisporum lineage A1/D3, V. isaacii, V. klebahnii, V. nonalfalfae, V. nubilum, V. tricorpus, V. zaregamsianum, and Species A1 and Species D1, the two undescribed ancestors of V. longisporum. The four multiple target (multiplex) PCR assays simultaneously differentiated the species or lineages within the following four groups: Verticillium albo-atrum, V. alfalfae and V. nonalfalfae; Verticillium dahliae and V. longisporum lineages A1/D1, A1/D2 and A1/D3; Verticillium dahliae including V. longisporum lineage A1/D3, V. isaacii, V. klebahnii and V. tricorpus; Verticillium isaacii, V. klebahnii and V. tricorpus. Since V. dahliae is a parent of two of the three lineages of the diploid hybrid V. longisporum, no simplex PCR assay is able to differentiate V. dahliae from all V. longisporum lineages. PCR assays were tested with fungal DNA extracts from pure cultures, and were not evaluated for detection and quantification of Verticillium species from plant or soil samples. The DNA sequence alignments are provided and can be used for the design of additional primers. PMID:23823707

  2. The ascomycete Verticillium longisporum is a hybrid and a plant pathogen with an expanded host range.

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    Patrik Inderbitzin

    Full Text Available Hybridization plays a central role in plant evolution, but its overall importance in fungi is unknown. New plant pathogens are thought to arise by hybridization between formerly separated fungal species. Evolution of hybrid plant pathogens from non-pathogenic ancestors in the fungal-like protist Phytophthora has been demonstrated, but in fungi, the most important group of plant pathogens, there are few well-characterized examples of hybrids. We focused our attention on the hybrid and plant pathogen Verticillium longisporum, the causal agent of the Verticillium wilt disease in crucifer crops. In order to address questions related to the evolutionary origin of V. longisporum, we used phylogenetic analyses of seven nuclear loci and a dataset of 203 isolates of V. longisporum, V. dahliae and related species. We confirmed that V. longisporum was diploid, and originated three different times, involving four different lineages and three different parental species. All hybrids shared a common parent, species A1, that hybridized respectively with species D1, V. dahliae lineage D2 and V. dahliae lineage D3, to give rise to three different lineages of V. longisporum. Species A1 and species D1 constituted as yet unknown taxa. Verticillium longisporum likely originated recently, as each V. longisporum lineage was genetically homogenous, and comprised species A1 alleles that were identical across lineages.

  3. Bacillus thuringiensis and Bacillus weihenstephanensis Inhibit the Growth of Phytopathogenic Verticillium Species

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    Hollensteiner, Jacqueline; Wemheuer, Franziska; Harting, Rebekka; Kolarzyk, Anna M.; Diaz Valerio, Stefani M.; Poehlein, Anja; Brzuszkiewicz, Elzbieta B.; Nesemann, Kai; Braus-Stromeyer, Susanna A.; Braus, Gerhard H.; Daniel, Rolf; Liesegang, Heiko

    2017-01-01

    Verticillium wilt causes severe yield losses in a broad range of economically important crops worldwide. As many soil fumigants have a severe environmental impact, new biocontrol strategies are needed. Members of the genus Bacillus are known as plant growth-promoting bacteria (PGPB) as well as biocontrol agents of pests and diseases. In this study, we isolated 267 Bacillus strains from root-associated soil of field-grown tomato plants. We evaluated the antifungal potential of 20 phenotypically diverse strains according to their antagonistic activity against the two phytopathogenic fungi Verticillium dahliae and Verticillium longisporum. In addition, the 20 strains were sequenced and phylogenetically characterized by multi-locus sequence typing (MLST) resulting in 7 different Bacillus thuringiensis and 13 Bacillus weihenstephanensis strains. All B. thuringiensis isolates inhibited in vitro the tomato pathogen V. dahliae JR2, but had only low efficacy against the tomato-foreign pathogen V. longisporum 43. All B. weihenstephanensis isolates exhibited no fungicidal activity whereas three B. weihenstephanensis isolates showed antagonistic effects on both phytopathogens. These strains had a rhizoid colony morphology, which has not been described for B. weihenstephanensis strains previously. Genome analysis of all isolates revealed putative genes encoding fungicidal substances and resulted in identification of 304 secondary metabolite gene clusters including 101 non-ribosomal polypeptide synthetases and 203 ribosomal-synthesized and post-translationally modified peptides. All genomes encoded genes for the synthesis of the antifungal siderophore bacillibactin. In the genome of one B. thuringiensis strain, a gene cluster for zwittermicin A was detected. Isolates which either exhibited an inhibitory or an interfering effect on the growth of the phytopathogens carried one or two genes encoding putative mycolitic chitinases, which might contribute to antifungal activities

  4. Bacillus thuringiensis and Bacillus weihenstephanensis Inhibit the Growth of Phytopathogenic Verticillium Species.

    Science.gov (United States)

    Hollensteiner, Jacqueline; Wemheuer, Franziska; Harting, Rebekka; Kolarzyk, Anna M; Diaz Valerio, Stefani M; Poehlein, Anja; Brzuszkiewicz, Elzbieta B; Nesemann, Kai; Braus-Stromeyer, Susanna A; Braus, Gerhard H; Daniel, Rolf; Liesegang, Heiko

    2016-01-01

    Verticillium wilt causes severe yield losses in a broad range of economically important crops worldwide. As many soil fumigants have a severe environmental impact, new biocontrol strategies are needed. Members of the genus Bacillus are known as plant growth-promoting bacteria (PGPB) as well as biocontrol agents of pests and diseases. In this study, we isolated 267 Bacillus strains from root-associated soil of field-grown tomato plants. We evaluated the antifungal potential of 20 phenotypically diverse strains according to their antagonistic activity against the two phytopathogenic fungi Verticillium dahliae and Verticillium longisporum. In addition, the 20 strains were sequenced and phylogenetically characterized by multi-locus sequence typing (MLST) resulting in 7 different Bacillus thuringiensis and 13 Bacillus weihenstephanensis strains. All B. thuringiensis isolates inhibited in vitro the tomato pathogen V. dahliae JR2, but had only low efficacy against the tomato-foreign pathogen V. longisporum 43. All B. weihenstephanensis isolates exhibited no fungicidal activity whereas three B. weihenstephanensis isolates showed antagonistic effects on both phytopathogens. These strains had a rhizoid colony morphology, which has not been described for B. weihenstephanensis strains previously. Genome analysis of all isolates revealed putative genes encoding fungicidal substances and resulted in identification of 304 secondary metabolite gene clusters including 101 non-ribosomal polypeptide synthetases and 203 ribosomal-synthesized and post-translationally modified peptides. All genomes encoded genes for the synthesis of the antifungal siderophore bacillibactin. In the genome of one B. thuringiensis strain, a gene cluster for zwittermicin A was detected. Isolates which either exhibited an inhibitory or an interfering effect on the growth of the phytopathogens carried one or two genes encoding putative mycolitic chitinases, which might contribute to antifungal activities

  5. Growth of Verticillium longisporum in Xylem Sap of Brassica napus is Independent from Cultivar Resistance but Promoted by Plant Aging.

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    Lopisso, Daniel Teshome; Knüfer, Jessica; Koopmann, Birger; von Tiedemann, Andreas

    2017-09-01

    As Verticillium stem striping of oilseed rape (OSR), a vascular disease caused by Verticillium longisporum, is extending into new geographic regions and no control with fungicides exists, the demand for understanding mechanisms of quantitative resistance increases. Because V. longisporum is strictly limited to the xylem and resistance is expressed in the systemic stage post root invasion, we investigated a potential antifungal role of soluble constituents and nutritional conditions in xylem sap as determinants of cultivar resistance of OSR to V. longisporum. Assessment of biometric and molecular genetic parameters applied to describe V. longisporum resistance (net area under disease progress curve, stunting, stem thickness, plant biomass, and V. longisporum DNA content) showed consistent susceptibility of cultivar 'Falcon' in contrast to two resistant genotypes, 'SEM' and 'Aviso'. Spectrophotometric analysis revealed a consistently stronger in vitro growth of V. longisporum in xylem sap extracted from OSR compared with the water control. Further comparisons of fungal growth in xylem sap of different cultivars revealed the absence of constitutive or V. longisporum induced antifungal activity in the xylem sap of resistant versus susceptible genotypes. The similar growth of V. longisporum in xylem sap, irrespective of cultivar, infection with V. longisporum and xylem sap filtration, was correlated with about equal amounts of total soluble proteins in xylem sap from these treatments. Interestingly, compared with younger plants, xylem sap from older plants induced significantly stronger fungal growth. Growth enhancement of V. longisporum in xylem sap of aging plants was reflected by increased contents of carbohydrates, which was consistent in mock or V. longisporum-infected plants and independent from cultivar resistance. The improved nutritional conditions in the xylem of more mature plants may explain the late appearance of disease symptoms, which are observed only in

  6. Verticillium longisporum infection induces organ-specific glucosinolate degradation in Arabidopsis thaliana

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    Katja eWitzel

    2015-07-01

    Full Text Available The species Verticillium represents a group of highly destructive fungal pathogens, responsible for vascular wilt in a number of crops. The host response to infection by V. longisporum at the level of secondary plant metabolites has not been well explored. Natural variation in the glucosinolate (GLS composition of four Arabidopsis thaliana accessions was characterized: the accessions Bur-0 and Hi-0 accumulated alkenyl GLS, while 3-hydroxypropyl GLS predominated in Kn-0 and Ler-0. With respect to GLS degradation products, Hi-0 and Kn-0 generated mainly isothiocyanates, whereas Bur-0 released epithionitriles and Ler-0 nitriles. An analysis of the effect on the composition of both GLS and its breakdown products in the leaf and root following the plants' exposure to V. longisporum revealed a number of organ- and accession-specific alterations. In the less disease susceptible accessions Bur-0 and Ler-0, colonization depressed the accumulation of GLS in the rosette leaves but accentuated it in the roots. In contrast, in the root, the level of GLS breakdown products in three of the four accessions fell, suggestive of their conjugation or binding to a fungal target molecule(s. The plant-pathogen interaction influenced both the organ- and accession-specific formation of GLS degradation products.

  7. The Vascular Pathogen Verticillium longisporum Does Not Affect Water Relations and Plant Responses to Drought Stress of Its Host, Brassica napus.

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    Lopisso, Daniel Teshome; Knüfer, Jessica; Koopmann, Birger; von Tiedemann, Andreas

    2017-04-01

    Verticillium longisporum is a host-specific vascular pathogen of oilseed rape (Brassica napus L.) that causes economic crop losses by impairing plant growth and inducing premature senescence. This study investigates whether plant damage through Verticillium stem striping is due to impaired plant water relations, whether V. longisporum affects responses of a susceptible B. napus variety to drought stress, and whether drought stress, in turn, affects plant responses to V. longisporum. Two-factorial experiments on a susceptible cultivar of B. napus infected or noninfected with V. longisporum and exposed to three watering levels (30, 60, and 100% field capacity) revealed that drought stress and V. longisporum impaired plant growth by entirely different mechanisms. Although both stresses similarly affected plant growth parameters (plant height, hypocotyl diameter, and shoot and root dry matter), infection of B. napus with V. longisporum did not affect any drought-related physiological or molecular genetic plant parameters, including transpiration rate, stomatal conductance, photosynthesis rate, water use efficiency, relative leaf water content, leaf proline content, or the expression of drought-responsive genes. Thus, this study provides comprehensive physiological and molecular genetic evidence explaining the lack of wilt symptoms in B. napus infected with V. longisporum. Likewise, drought tolerance of B. napus was unaffected by V. longisporum, as was the level of disease by drought conditions, thus excluding a concerted action of both stresses in the field. Although it is evident that drought and vascular infection with V. longisporum impair plant growth by different mechanisms, it remains to be determined by which other factors V. longisporum causes crop loss.

  8. Verticillium longisporum Infection Affects the Leaf Apoplastic Proteome, Metabolome, and Cell Wall Properties in Arabidopsis thaliana

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    Floerl, Saskia; Majcherczyk, Andrzej; Possienke, Mareike; Feussner, Kirstin; Tappe, Hella; Gatz, Christiane; Feussner, Ivo; Kües, Ursula; Polle, Andrea

    2012-01-01

    Verticillium longisporum (VL) is one of the most devastating diseases in important oil crops from the family of Brassicaceae. The fungus resides for much time of its life cycle in the extracellular fluid of the vascular system, where it cannot be controlled by conventional fungicides. To obtain insights into the biology of VL-plant interaction in the apoplast, the secretome consisting of the extracellular proteome and metabolome as well as cell wall properties were studied in the model Brassicaceae, Arabidopsis thaliana. VL infection resulted in increased production of cell wall material with an altered composition of carbohydrate polymers and increased lignification. The abundance of several hundred soluble metabolites changed in the apoplast of VL-infected plants including signalling and defence compounds such as glycosides of salicylic acid, lignans and dihydroxybenzoic acid as well as oxylipins. The extracellular proteome of healthy leaves was enriched in antifungal proteins. VL caused specific increases in six apoplast proteins (three peroxidases PRX52, PRX34, P37, serine carboxypeptidase SCPL20, α-galactosidase AGAL2 and a germin-like protein GLP3), which have functions in defence and cell wall modification. The abundance of a lectin-like, chitin-inducible protein (CILLP) was reduced. Since the transcript levels of most of the induced proteins were not elevated until late infection time points (>20 dpi), whereas those of CILLP and GLP3 were reduced at earlier time points, our results may suggest that VL enhances its virulence by rapid down-regulation and delay of induction of plant defence genes. PMID:22363647

  9. RabGAP22 is required for defense to the vascular pathogen Verticillium longisporum and contributes to stomata immunity.

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    Jonas Roos

    Full Text Available Verticillium longisporum is a soil-borne pathogen with a preference for plants within the family Brassicaceae. Following invasion of the roots, the fungus proliferates in the plant vascular system leading to stunted plant growth, chlorosis and premature senescence. RabGTPases have been demonstrated to play a crucial role in regulating multiple responses in plants. Here, we report on the identification and characterization of the Rab GTPase-activating protein RabGAP22 gene from Arabidopsis, as an activator of multiple components in the immune responses to V. longisporum. RabGAP22Pro :GUS transgenic lines showed GUS expression predominantly in root meristems, vascular tissues and stomata, whereas the RabGAP22 protein localized in the nucleus. Reduced RabGAP22 transcript levels in mutants of the brassinolide (BL signaling gene BRI1-associated receptor kinase 1, together with a reduction of fungal proliferation following BL pretreatment, suggested RabGAP22 to be involved in BL-mediated responses. Pull-down assays revealed serine:glyoxylate aminotransferase (AGT1 as an interacting partner during V. longisporum infection and bimolecular fluorescence complementation (BiFC showed the RabGAP22-AGT1 protein complex to be localized in the peroxisomes. Further, fungal-induced RabGAP22 expression was found to be associated with elevated endogenous levels of the plant hormones jasmonic acid (JA and abscisic acid (ABA. An inadequate ABA response in rabgap22-1 mutants, coupled with a stomata-localized expression of RabGAP22 and impairment of guard cell closure in response to V. longisporum and Pseudomonas syringae, suggest that RabGAP22 has multiple roles in innate immunity.

  10. Transposable elements in phytopathogenic Verticillium spp.: insights into genome evolution and inter- and intra-specific diversification

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    Amyotte Stefan G

    2012-07-01

    Full Text Available Abstract Background Verticillium dahliae (Vd and Verticillium albo-atrum (Va are cosmopolitan soil fungi causing very disruptive vascular diseases on a wide range of crop plants. To date, no sexual stage has been identified in either microorganism suggesting that somatic mutation is a major force in generating genetic diversity. Whole genome comparative analysis of the recently sequenced strains VdLs.17 and VaMs.102 revealed that non-random insertions of transposable elements (TEs have contributed to the generation of four lineage-specific (LS regions in VdLs.17. Results We present here a detailed analysis of Class I retrotransposons and Class II “cut-and-paste” DNA elements detected in the sequenced Verticillium genomes. We report also of their distribution in other Vd and Va isolates from various geographic origins. In VdLs.17, we identified and characterized 56 complete retrotransposons of the Gypsy-, Copia- and LINE-like types, as well as 34 full-length elements of the “cut-and-paste” superfamilies Tc1/mariner, Activator and Mutator. While Copia and Tc1/mariner were present in multiple identical copies, Activator and Mutator sequences were highly divergent. Most elements comprised complete ORFs, had matching ESTs and showed active transcription in response to stress treatment. Noticeably, we found evidences of repeat-induced point mutation (RIP only in some of the Gypsy retroelements. While Copia-, Gypsy- and Tc1/mariner-like transposons were prominent, a large variation in presence of the other types of mobile elements was detected in the other Verticillium spp. strains surveyed. In particular, neither complete nor defective “cut-and-paste” TEs were found in VaMs.102. Conclusions Copia-, Gypsy- and Tc1/mariner-like transposons are the most wide-spread TEs in the phytopathogens V. dahliae and V. albo-atrum. In VdLs.17, we identified several retroelements and “cut-and-paste” transposons still potentially active. Some of these

  11. "Cryptic" group-I introns in the nuclear SSU-rRNA gene of Verticillium dahliae.

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    Papaioannou, Ioannis A; Dimopoulou, Chrysoula D; Typas, Milton A

    2014-08-01

    Group-I introns are widespread--though irregularly distributed--in eukaryotic organisms, and they have been extensively used for discrimination and phylogenetic analyses. Within the Verticillium genus, which comprises important phytopathogenic fungi, a group-I intron was previously identified in the SSU-rRNA (18S) gene of only V. longisporum. In this work, we aimed at elucidating the SSU-located intron distribution in V. dahliae and other Verticillium species, and the assessment of heterogeneity regarding intron content among rDNA repeats of fungal strains. Using conserved PCR primers for the amplification of the SSU gene, a structurally similar novel intron (sub-group IC1) was detected in only a few V. dahliae isolates. However, when intron-specific primers were used for the screening of a diverse collection of Verticillium isolates that originally failed to produce intron-containing SSU amplicons, most were found to contain one or both intron types, at variable rDNA repeat numbers. This marked heterogeneity was confirmed with qRT-PCR by testing rDNA copy numbers (varying from 39 to 70 copies per haploid genome) and intron copy ratios in selected isolates. Our results demonstrate that (a) IC1 group-I introns are not specific to V. longisporum within the Verticillium genus, (b) V. dahliae isolates of vegetative compatibility groups (VCGs) 4A and 6, which bear the novel intron at most of their rDNA repeats, are closely related, and (c) there is considerable intra-genomic heterogeneity for the presence or absence of introns among the ribosomal repeats. These findings underline that distributions of introns in the highly heterogeneous repetitive rDNA complex should always be verified with sensitive methods to avoid misleading conclusions for the phylogeny of fungi and other organisms.

  12. Functional analysis of the tomato Ve resistance locus against Verticillium wilt

    NARCIS (Netherlands)

    Fradin, E.F.

    2011-01-01

    Verticillium dahliae, V. albo-atrum and V. longisporum are soil-borne plant pathogens that are responsible for Verticillium wilt diseases in temperate and subtropical regions. Collectively they can infect over 200 hosts, including many economically important crops. Chapter 1 is a “pathogen pro

  13. Genetic dissection of Verticillium wilt resistance mediated by tomato Ve1

    NARCIS (Netherlands)

    Fradin, E.F.; Zhang, Z.; Juarez Ayala, J.C.; Castroverde, C.C.M.; Nazar, R.N.; Robb, J.; Liu, Chun-Ming; Thomma, B.P.H.J.

    2009-01-01

    Vascular wilt diseases caused by soil-borne pathogens are among the most devastating plant diseases worldwide. The Verticillium genus includes vascular wilt pathogens with a wide host range. Although V. longisporum infects various hosts belonging to the Cruciferaceae, V. dahliae and V. albo-atrum ca

  14. Analysis of a MULE-cyanide hydratase gene fusion in Verticillium dahliae

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    The genome of the phytopathogenic fungus Verticillium dahliae encodes numerous Class II “cut-and-paste” transposable elements, including those of a small group of MULE transposons. We have previously identified a fusion event between a MULE transposon sequence and sequence encoding a cyanide hydrata...

  15. Biosynthetic arginine decarboxylase in phytopathogenic fungi.

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    Khan, A J; Minocha, S C

    1989-01-01

    It has been reported that while bacteria and higher plants possess two different pathways for the biosynthesis of putrescine, via ornithine decarboxylase (ODC) and arginine decarboxylase (ADC); the fungi, like animals, only use the former pathway. We found that contrary to the earlier reports, two of the phytopathogenic fungi (Ceratocystis minor and Verticillium dahliae) contain significant levels of ADC activity with very little ODC. The ADC in these fungi has high pH optimum (8.4) and low Km (0.237 mM for C. minor, 0.103 mM for V. dahliae), and is strongly inhibited by alpha-difluoromethylarginine (DFMA), putrescine and spermidine, further showing that this enzyme is probably involved in the biosynthesis of polyamines and not in the catabolism of arginine as in Escherichia coli. The growth of these fungi is strongly inhibited by DFMA while alpha-difluoromethylornithine (DFMO) has little effect.

  16. Control of Verticillium Yellows in Chinese Cabbage by the Dark Septate Endophytic Fungus LtVB3.

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    Narisawa, K; Usuki, F; Hashiba, T

    2004-05-01

    ABSTRACT Three hundred forty-nine fungal endophytes were obtained from a total of 1,214 root segments of eggplant, melon, barley, and Chinese cabbage grown as bait plants in a mixed soil made up of samples from different forest soils in Alberta and British Columbia, Canada. Three of the 349 isolates, when inoculated in axenically reared Chinese cabbage seedlings grown in petri dishes, almost completely suppressed the effects of a postinoculated and virulent strain of Verticillium longisporum. Two isolates effective against the pathogen were Phialocephala fortinii, which had been obtained from the roots of eggplant and Chinese cabbage. The third isolate was a dark septate endophytic (DSE) fungus obtained from barley roots. Hyphae of P. fortinii grew along the surface of the root and formed microsclerotia on or in the epidermal layer. Hyphae of the DSE fungus heavily colonized root cells of the cortex. Seedlings grown for 1 week in the presence of the endophytes were then challenged with the Verticillium pathogen. In DSE-treated roots, some of cell walls in the epidermal and cortical layers showed cell wall appositions and thickenings, which appeared to limit the ingress of the pathogen into adjacent cells. Such marked host reactions were not observed in the root cells colonized by P. fortinii. Chinese cabbage preinoculated with the above endophytes and, for comparison, a previously reported disease-suppressive fungal endophyte, Heteroconium chaetospira, were transplanted into the field and disease symptoms were assessed. The DSE could most effectively inhibit the development of Verticillium yellows, with reductions in the percentages of external and internal disease symptoms of 84 and 88%, respectively. The protective values against the disease are extremely high compared with those of other isolates. Most of the DSE-treated plants in the plots achieved marketable quality.

  17. Genetic dissection of Verticillium wilt resistance mediated by tomato Ve1.

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    Fradin, Emilie F; Zhang, Zhao; Juarez Ayala, Juan C; Castroverde, Christian D M; Nazar, Ross N; Robb, Jane; Liu, Chun-Ming; Thomma, Bart P H J

    2009-05-01

    Vascular wilt diseases caused by soil-borne pathogens are among the most devastating plant diseases worldwide. The Verticillium genus includes vascular wilt pathogens with a wide host range. Although V. longisporum infects various hosts belonging to the Cruciferaceae, V. dahliae and V. albo-atrum cause vascular wilt diseases in over 200 dicotyledonous species, including economically important crops. A locus responsible for resistance against race 1 strains of V. dahliae and V. albo-atrum has been cloned from tomato (Solanum lycopersicum) only. This locus, known as Ve, comprises two closely linked inversely oriented genes, Ve1 and Ve2, that encode cell surface receptor proteins of the extracellular leucine-rich repeat receptor-like protein class of disease resistance proteins. Here, we show that Ve1, but not Ve2, provides resistance in tomato against race 1 strains of V. dahliae and V. albo-atrum and not against race 2 strains. Using virus-induced gene silencing in tomato, the signaling cascade downstream of Ve1 is shown to require both EDS1 and NDR1. In addition, NRC1, ACIF, MEK2, and SERK3/BAK1 also act as positive regulators of Ve1 in tomato. In conclusion, Ve1-mediated resistance signaling only partially overlaps with signaling mediated by Cf proteins, type members of the receptor-like protein class of resistance proteins.

  18. A novel method for rapidly isolating microbes that suppress soil-borne phytopathogens

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    Cooper, Sarah; Agnew, Linda; Pereg, Lily

    2016-04-01

    Seedling establishment faces a large number of challenges related to soil physical properties as well as to fungal root diseases. It is extremely difficult to eliminate fungal pathogens from soils where their populations are established due to the persistent nature of their spores and since fumigation of resident fungi is very ineffective in clay-containing soils. Therefore it is necessary to find ways to overcome disease in areas where the soils are infected with fungal phytopathogens. The phenomenon of disease suppressive soils, where the pathogen is present but no disease observed, suggests that microbial antagonism in the soil may lead to the suppression of the growth of fungal pathogens. There are also cases in the literature where soil microorganisms were isolated that suppress the growth of phytopathogens. Antibiosis is one of the most important mechanisms responsible for fungal antagonism, with some significant antifungal compounds involved including antibiotics, volatile organic compounds, hydrogen cyanide and lytic enzymes. Isolation of pathogen-suppressive microorganisms from the soil is time consuming and tedious. We established a simple method for direct isolation of soil microbes (bacteria and fungi) that suppress fungal phytopathogens as well as procedures for confirmation of disease suppression. We will discuss such methods, which were so far tested with the cotton fungal pathogens Thielaviopsis basicola, Verticillium dahliae and Fusarium oxysporum and Verticillium fungicola. We have isolated a diversity of T. basicola-suppressive fungi and bacteria from two vastly different soil types. Identification of the antagonistic isolates revealed that they are a diverse lot, some belong to groups known to be suppressive of a wide range of fungal pathogens, endorsing the power of this technique to rapidly and directly isolate soil-borne microbes antagonistic to a wide variety of fungal pathogens.

  19. Expression of BvGLP-1 encoding a germin-like protein from sugar beet in Arabidopsis thaliana leads to resistance against phytopathogenic fungi.

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    Knecht, Katrin; Seyffarth, Monique; Desel, Christine; Thurau, Tim; Sherameti, Irena; Lou, Binggan; Oelmüller, Ralf; Cai, Daguang

    2010-04-01

    Nematode (Heterodera schachtii) resistance in sugar beet (Beta vulgaris) is controlled by a single dominant resistance gene, Hs1(pro-1). BvGLP-1 was cloned from resistant sugar beet. The BvGLP-1 messenger (m)RNA is highly upregulated in the resistant plants after nematode infection, suggesting its role in the Hs1(pro-1) mediated resistance. BvGLP-1 exhibits sequence homology to a set of plant germin-like proteins (GLP), from which several have proved to be functional in plant basal or defense resistance against fungal pathogens. To test whether BvGLP-1 is also involved in the plant-fungus interaction, we transferred BvGLP-1 into Arabidopsis and challenged the transgenic plants with the pathogenic fungi Verticillium longisporum and Rhizoctonia solani as well as with the beneficial endophytic fungus Piriformospora indica. The expression of BvGLP-1 in Arabidopsis elevated the H(2)O(2) content and conferred significant resistance to V. longisporum and R. solani but did not affect the beneficial interaction with P. indica in seedlings. Microscopic observations revealed a dramatic reduction in the amount of hyphae of the pathogenic fungi on the root surface as well as of fungal mycelium developed inside the roots of transgenic Arabidopsis compared with wild-type plants. Molecular analysis demonstrated that the BvGLP-1 expression in Arabidopsis constitutively activates the expression of a subset of plant defense-related proteins such as PR-1 to PR-4 and PDF1.2 but not PDF2.1 and PDF2.3. In contrast, the PDF2.1 mRNA level was downregulated. These data suggest an important role of BvGLP-1 in establishment of plant defense responses, which follow specific signaling routes that diverge from those induced by the beneficial fungus.

  20. Biological control agent against verticillium wilt

    CSIR Research Space (South Africa)

    Gumede, WHN

    2006-04-01

    Full Text Available Verticillium wilt Presented at the 14th Biennial congress of the South African Society for Microbiology WHN Gumede CSIR Biosciences 11-04-06 Slide 2 © CSIR 2006 www.csir.co.za INTRODUCTION • Historical background • Application...

  1. Ecology and biological control of Verticillium dahliae

    NARCIS (Netherlands)

    Soesanto, L.

    2000-01-01

    The dynamics of Verticillium dahliae , the causal agent of wilt disease in many crops including potato, cotton, and olive, were investigated. Its biological control with Talaromyces flavus with or without additional Pseudomonas fluorescens was attempted. Arabidopsis thaliana was selected as a bioass

  2. Verticillium wilt in trees. Detection, prediction and disease management

    NARCIS (Netherlands)

    Goud, J.C.

    2003-01-01

    Verticillium dahliae , the causal agent of verticillium wilt, is the cause of high losses in a number of crops, especially nursery trees. Existing methods for quantification of V. dahliae microsclerotia in the soil were compared. The distinction of V. dahliae and V. tricorpus was studied on two semi

  3. QTL mapping and candidate gene discovery in potato for resistance to the Verticillium wilt pathogen Verticillium dahliae

    Science.gov (United States)

    Verticillium wilt (VW) of potato (Solanum tuberosum), caused by fungal pathogens, Verticillium dahliae and V. albo atrum, is a disease of major significance throughout the potato growing regions in the world. In the past, researchers have focused on the Ve gene, which is a major dominant gene that c...

  4. First report of Verticillium wilt caused by Verticillium nonalfalfae on tree-of-heaven (Ailanthus altissima) in Ohio

    Science.gov (United States)

    J. Rebbeck; M.A. Malone; D.P.G. Short; M.T. Kasson; E.S. O' Neal; D.D. Davis

    2013-01-01

    Verticillium wilt of the highly invasive tree-of-heaven [Ailanthus altissima (Mill.) Swingle], caused by Verticillium nonalfalfae Interbitzin et al. (1), formerly classified as V. albo-atrum Reinke and Berthold, has been reported in the United States from two states: Pennsylvania (2) and Virginia (3). Infected...

  5. Desirable Traits of a Good Biocontrol Agent against Verticillium Wilt.

    Science.gov (United States)

    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

  6. Desirable Traits of a Good Biocontrol Agent against Verticillium Wilt

    Directory of Open Access Journals (Sweden)

    Silke Deketelaere

    2017-07-01

    Full Text Available 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

  7. Distribution of phytopathogenic bacteria in infested seeds

    Science.gov (United States)

    Populations of phytopathogenic bacteria representing five host-pathogen combinations were assessed to determine if there was a mathematical relationship common across seedborne bacterial diseases. Bacterial populations were estimated from naturally-infested seeds of cowpea (Vigna unguiculata), peppe...

  8. Molecular determinants of resistance to Verticillium dahliae in potato

    Science.gov (United States)

    A constant evolutionary arms race between host resistance genes and pathogen effectors determine adaptive fitness. Therefore, identification of both host resistance genes and pathogen effectors is important in devising effective strategies to control disease. In tomato, resistance to Verticillium da...

  9. Identification and characterization of a pathogenicity-related gene VdCYP1 from Verticillium dahliae

    Science.gov (United States)

    Zhang, Dan-Dan; Wang, Xin-Yan; Chen, Jie-Yin; Kong, Zhi-Qiang; Gui, Yue-Jing; Li, Nan-Yang; Bao, Yu-Ming; Dai, Xiao-Feng

    2016-01-01

    Verticillium dahliae is a phytopathogenic fungus that causes vascular wilt disease in a wide variety of crop plants, thereby causing extensive economic loss. In present study, one V. dahliae T-DNA mutant M01C06 showed the pathogenicity loss on cotton, and the expression of a flanking gene encoding cytochrome P450 monooxygenase (P450, VdCYP1) was strongly repressed. P450s of fungi could affect the fungal pathogenicity by involving in the synthesis of secondary metabolites. However, there was no report about the pathogenic function of P450s in V. dahliae. VdCYP1 gene deletion and complementation experiments confirmed that VdCYP1 was the pathogenicity-related gene in V. dahliae. A comparison of culture supernatants of the VdCYP1 deletion mutants and wild-type strains indicates that at least 14 kinds of secondary metabolites syntheses were affected due to VdCYP1 gene deletion. One of these compounds, sulfacetamide, had the ability to induce the necrosis and wilting symptoms in cotton. Above results indicate that VdCYP1 could participate in pathogenesis by involving the secondary metabolism in V. dahliae, such as the compound sulfacetamide. In conclusion, VdCYP1 acts as an important pathogenicity-related factor to involve in secondary metabolism that likely contributes to the pathogenic process in V. dahliae. PMID:27329129

  10. Differentiation of Phytopathogenic Agrobacterium spp.

    Directory of Open Access Journals (Sweden)

    Nemanja Kuzmanović

    2011-01-01

    Full Text Available Due to the difficulties in differentiation of phytopathogenic Agrobacterium spp. and lack of a standardized protocol, we carried out selection and evaluation of suitable methods based on the bacterial physiological, genetic and pathogenic properties. Strains of Agrobacterium tumefaciens, A. rhizogenes and A. vitis were differentiated using standard bacteriological and molecular methods. The biochemical and physiological tests confirmed authenticity of the strains. Two duplex PCR methods were conducted with four different primer pairs. In all strains, presence of plasmid virD2 and virC pathogenicity genes was detected. Chromosomal pehA gene was determined in A. vitis strain. Pathogenicity was confirmed on carrot slices and young plants of tomato and sunflower. Strains of A. tumefaciens and A. vitis were pathogenic on all test plants, while strain of A. rhizogenes induced characteristic symptoms only on carrot slices. The tests used in this study provided reliable discrimination between the three species and confirmed their identity as tumorigenic (TiAgrobacterium tumefaciens and A. vitis, and rhizogenic (Ri A. rhizogenes.

  11. Verticillium wilt in nursery trees: damage thresholds, spatial and temporal aspects

    NARCIS (Netherlands)

    Goud, J.C.; Termorshuizen, A.J.; Bruggen, van A.H.C.

    2011-01-01

    Verticillium wilt can cause high losses in tree nurseries. To be able to predict disease and unravel disease dynamics over time and space, the relationship between verticillium wilt and soil inoculum densities of Verticillium dahliae and the nematode Pratylenchus fallax was studied in two 4-year fie

  12. Molecular research and genetic engineering of resistance to Verticillium wilt in cotton: A review

    Science.gov (United States)

    Verticillium dahliae, a soil-borne pathogen, causes Verticillium wilt, one of the most serious diseases in cotton, deleteriously influencing the crop’s production and quality. Verticillium wilt has become a major obstacle in cotton production since Helicoverpa armigera, the cotton bollworm, became e...

  13. Interfamily Transfer of Tomato Ve1 Mediates Verticillium Resistance in Arabidopsis

    NARCIS (Netherlands)

    Fradin, E.F.; Abd-El-Haliem, A.; Masini, L.; Berg, van den G.C.M.; Joosten, M.H.A.J.; Thomma, B.P.H.J.

    2011-01-01

    Vascular wilts caused by soil-borne fungal species of the Verticillium genus are devastating plant diseases. The most common species, Verticillium dahliae and Verticillium albo-atrum, have broad host ranges and are notoriously difficult to control. Therefore, genetic resistance is the preferred meth

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

    Directory of Open Access Journals (Sweden)

    Gabriel Moura Mascarin

    2016-09-01

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

  15. Potential antagonism of some Trichoderma strains isolated from Moroccan soil against three phytopathogenic fungi of great economic importance

    Directory of Open Access Journals (Sweden)

    Wafaa MOKHTARI

    2017-09-01

    Full Text Available In this study, 17 Trichoderma strains were isolated from different soils (crop fields and Argan forests in Morocco. Purified monospore cultures were identified using molecular methods and tested for their potential antagonism against three phytopathogenic fungi (Fusarium oxyxporum, verticillium dahlia and rhizoctonia solani. After DNA extraction, translation elongation factor (tef1 was amplified in extracts of 17 strains, sequenced and compared with their ex-types. As a result, three species were identified among the strains, which clustered in two different subclades of Trichoderma: the species T. afroharzianum, and T. guizhouense belong to the Harzianum clade, while T. longibrachiatum belongs to the Longibrachiatum clade. Investigation of potential antagonistic effects of these strains against the soil-borne phytopathogens F. oxysporum, R. solani and V. dahliae was conducted in a dual culture plate assay, using 17 promising Trichoderma strains that have been selected based on a polymerase chain reaction (PCR screening approach. In vitro, Trichoderma isolates showed effective antagonistic performance by decreasing soil borne pathogens mycelium radial growth. Trichoderma afroharzianum showed the highest Percentage of Radial Inhibition Growth (PRIG %. The highest PRIG% = 98% was for 8A2.3 isolate against R. solani and the lowest PRIG%= 67% for T9i10 against F. oxysporum. On the other hand, T9i12, which is T. reesei species, led to a high radial inhibition of pathogens’ mycelium.

  16. [Antagonistic interactions between saprotrophic fungi and geohelminths. 1. Saprotrophic fungi in the biological control of phytopathogenic geohelminths].

    Science.gov (United States)

    Mazurkiewicz-Zapałowicz, Kinga; Kołodziejczyk, Lidia

    2009-01-01

    The state of knowledge on the possible antagonism between soil saprotrophic fungi and phytopathogenic nematodes of the genera Meloidogyne, Heterodera, and Globodera is reviewed basing on the literature and our own research. Mycelial colonisation of various developmental stages of these geohelminths is the most common factor thought to reduce their populations in nature. The following parasitic fungi can be found on the cysts, eggs, as well as the larvae of the nematodes: Paecilomyces lilacinus, Verticillium chlamydosporium, Cylindrocarpon destructans, Pochonia chlamydosporia, Fusarium spp., and Penicillium spp. The fungi invade the nematodes, such as Heterodera, Globodera, or Meloidogyne, "passively" penetrating through the natural orifices of the cysts, eggs, and larvae of the host. Equally frequent, however, is a biochemical action of the fungi prior to colonisation, which is linked with production of mycotoxirls or hydrolytic enzymes. Such an active way of fungal penetration of various stages of the phytopathogenic nematodes has been observed in Pochonia chlamydosporia, Penicillium verrucosum var. cyclopium, P. frequentans, Sclerotinia rolfsii, Rhizoctonia solani, and Fusarium spp. Triacylglycerols (TAG), phenols, as well as trichothecene, T-2, have been found in the metabolites extracted from mycelia of these species. Predation by fungi is also a factor that may reduce a population of phytopathogenic nematodes. This form of antagonism is characteristic for nematicidal fungi of the genera Arthrobotrys and Dactylella. These fungi form shrinking rings and hooks in their mycelia by which the fungus entangles and paralyses a migrating form of nematode. Despite the fact that the antagonism between fungi and nematodes is a commonly occurring phenomenon observed in the soil, the nematicidal and nematotoxic properties of fungi have not a wide application in biological plant protection. Up till now, only the bionematicides based on Arthrobotrys robusta (Royal 300 and

  17. A synthetic antimicrobial peptide BTD-S expressed in Arabidopsis thaliana confers enhanced resistance to Verticillium dahliae.

    Science.gov (United States)

    Li, Feng; Shen, Hao; Wang, Ming; Fan, Kai; Bibi, Noreen; Ni, Mi; Yuan, Shuna; Wang, Xuede

    2016-08-01

    BTD-S is a synthetic non-cyclic θ-defensin derivative which was previously designed in our laboratory based on baboon θ-defensins (BTDs). It shows robust antimicrobial activity against economically important phytopathogen, Verticillium dahliae. Here, we deduced the coding nucleotide sequence of BTD-S and introduced the gene into wild-type (ecotype Columbia-0) Arabidopsis thaliana plants. Results demonstrated that BTD-S-transgenic lines displayed in bioassays inhibitory effects on the growth of V. dahliae in vivo and in vitro. Based on symptom severity, enhanced resistance was found in a survey of BTD-S-transgenic lines. Besides, crude protein extracts from root tissues of BTD-S-transformed plants significantly restricted the growth of fungal hyphae and the germination of conidia. Also, fungal biomass over time determined by real-time PCR demonstrated the overgrowth of V. dahliae in wild-type plants 2-3 weeks after inoculation, while almost no fungal DNA was detected in aerial tissues of their transgenic progenitors. The result suggested that fungus failed to invade and progress acropetally up to establish a systemic infection in BTD-S-transgenic plants. Moreover, the assessment of basal defense responses was performed in the leaves of WT and BTD-S-transgenic plants. The mitigated oxidative stress and low antioxidase level in BTD-S-transgenic plants revealed that BTD-S acts via permeabilizing target microbial membranes, which is in a category different from hypersensitive response-dependent defense. Taken together, our results demonstrate that BTD-S is a promising gene to be explored for transgenic engineering for plant protection against Verticillium wilt.

  18. The Mitogen-Activated Protein Kinase Kinase VdPbs2 of Verticillium dahliae Regulates Microsclerotia Formation, Stress Response, and Plant Infection

    Science.gov (United States)

    Tian, Longyan; Wang, Yonglin; Yu, Jun; Xiong, Dianguang; Zhao, Hengjun; Tian, Chengming

    2016-01-01

    Verticillium dahliae, a ubiquitous phytopathogenic fungus, forms resting structures, known as microsclerotia that play crucial roles in Verticillium wilt diseases. VdHog1, a mitogen-activated protein kinase (MAPK), controls microsclerotia formation, virulence, and stress response in V. dahliae. In this study, we present detailed evidence that the conserved upstream component of VdHog1, VdPbs2, is a key regulator of microsclerotia formation, oxidative stress and fungicide response and plant virulence in V. dahliae. We identified VdPbs2, homologous to the yeast MAPK kinase Pbs2. Similar to the VdHog1 deletion mutant, VdPbs2 deletion strains exhibited delayed melanin synthesis and reduced formation of microsclerotia. When exposed to stresses, VdPbs2 mutants were more sensitive than the wild type to osmotic agents and peroxide, but more resistant to inhibitors of cell wall synthesis and some fungicides. Finally, VdPbs2 deletion mutants exhibited reduced virulence on smoke tree and tobacco seedlings. When taken together, we implicate that VdPbs2 and VdHog1 function in a cascade that regulates microsclerotia formation and virulence, but not all VdHog1 dependent functions are VdPbs2 regulated. This study thus provides novel insights into the signal transduction mechanisms that regulate microsclerotia formation and pathogenesis in this fungus. PMID:27729908

  19. Antimycotic activities of selected plant flora, growing wild in Lebanon, against phytopathogenic fungi.

    Science.gov (United States)

    Abou-Jawdah, Yusuf; Sobh, Hana; Salameh, Abdu

    2002-05-22

    Petroleum ether (PE) and methanolic extracts of nine wild plant species were tested in vitro for their antimycotic activity against eight phytopathogenic fungi. The efficacy of PE extracts against all pathogens tested was higher than that of methanolic extracts. Wild marjoram (Origanum syriacum) PE extract showed the highest and widest range of activity. It resulted in complete inhibition of mycelial growth of six of eight fungi tested and also gave nearly complete inhibition of spore germination of the six fungi included in the assay, namely, Botrytis cinerea, Alternaria solani, Penicillium sp., Cladosporium sp., Fusarium oxysporum f. sp. melonis, and Verticillium dahlia. The other plant extracts showed differential activities in the spore germination test, but none was highly active against mycelial growth. Inula viscosa and Mentha longifolia were highly effective (>88%) in spore germination tests against five of six fungi tested, whereas Centaurea pallescens, Cichorium intybus, Eryngium creticum, Salvia fruticosa, and Melia azedarach showed >95% inhibition of spore germination in at least two fungi. Foeniculum vulgare showed the least antimycotic activity. Fractionation followed by autobiography on TLC plates using Cladosporium sp. as a test organism showed that O. syriacum PE extracts contained three inhibition zones, and those of Inula viscosa and Cichorium intybus, two, whereas the PE extracts of the remaining plants showed each one inhibition zone. Some of the major compounds present in these inhibition zones were identified by GC-MS. The possibility for using these extracts, or their mixtures, to control plant diseases is discussed.

  20. Repetitive elements, architects of genomic variation in Verticillium

    Science.gov (United States)

    Vascular wilt pathogens in the genus Verticillium show considerable variation with respect to their host ranges, genomic organization, and the variety and number of transposable elements (TEs) that they carry. These families of TE sequences were first documented in the wide host range, plant pathog...

  1. Insertional mutagenesis in the vascular wilt pathogen Verticillium dahliae

    NARCIS (Netherlands)

    Santhanam, P.

    2014-01-01

    Vascular wilt diseases caused by soil-borne pathogens are among the most devastating plant diseases worldwide. The ascomycete fungus Verticillium dahliae causes vascular wilt diseases in hundreds of dicotyledonous plant species, including important crops such as eggplant, lettuce, olive, spinach

  2. Effect of solarization with fresh chicken manure on verticillium wilt ...

    African Journals Online (AJOL)

    BİM

    2016-12-28

    Dec 28, 2016 ... possible to use solar energy instead of fumigation, and it is also beneficial to use it ...... be overcome by supplementation of green plant or .... Verticillium wilt on resistant tomato cultivars in California: Virulence of isolates from ...

  3. BIOLOGICAL CONTRAOL OF PHYTOPATHOGENS USING ANTAGONIST TRICHODERMA VIRIDE

    OpenAIRE

    D.S.R. RAJENDRA SINGH; SHAIK SAYEED; K. BRUNDA EVI; B. BHADRAIAH

    2006-01-01

    Antagonistic fungus i.e. Trichoderma viride was tested in vitro against seven phytopathogens viz., Aspergillus niger, A. fumigatus, Macrophimina phaseolina, Fusarium oxysporum, F. solani, Paecilomyces varoti and sclerotium rolfsii. Trichoderma viride exhibited the antagonistic effect against these phytopathogens. Under dual culture the hyphal growth of the phytopathogens was inhibited at the zone of contact with the hyphae of the antagonist. Microscopic examination revealed that hyphal tips o...

  4. Selection for resistance to Verticillium wilt caused by race 2 isolates of Verticillium dahliae in accessions of lettuce (Lactuca sativa L.).

    Science.gov (United States)

    Verticillium wilt of lettuce caused by Verticillium dahliae can cause severe economic damage to lettuce producers. The pathogen exists as two races (races 1 and 2) in lettuce, and complete resistance to race 1 is known. Resistance to race 2 isolates has not been reported, and production of race 1 re...

  5. Iceberg lettuce breeding lines with resistance to Verticillium wilt caused by race 1 isolates of Verticillium dahliae.

    Science.gov (United States)

    The Agricultural Research Service, United States Department of Agriculture and the University of California, Davis, announce the release of two breeding lines of lettuce (Lactuca sativa L.). Lines RH08-0472 and RH08-0475 are F9 iceberg type lettuce breeding lines with resistance to Verticillium wil...

  6. Genomics spurs rapid advances in our understanding of the biology of vascular wilt pathogens in the genus Verticillium

    NARCIS (Netherlands)

    Klimes, A.; Dobinson, K.F.; Thomma, B.; Klosterman, S.J.

    2015-01-01

    The availability of genomic sequences of several Verticillium species triggered an explosion of genome-scale investigations of mechanisms fundamental to the Verticillium life cycle and disease process. Comparative genomics studies have revealed evolutionary mechanisms, such as hybridization and inte

  7. Resistance to Verticillium dahliae (Kleb.) in the strawberry breeding lines.

    Science.gov (United States)

    Zebrowska, J; Hortyński, J; Cholewa, T; Honcz, K

    2006-01-01

    Verticillium species are soil-borne fungi with worldwide distribution, causing vascular disease that results in severe yield and quality losses in fruit and nut crops, legumes, vegetables, forest trees, and woody and herbaceous ornamentals. Most crop diseases are caused by the two species Verticillium dahliae Klebahn and V. albo-atrum Reinke and Berthier, which differ in morphology, host range, and growth characteristics. The control of Verticillium spp. is especially difficult because they can survive in the soil as resting structures for several years. Cultivation of resistant plant material is the most effective method of the disease elimination. Resistance to Verticillium dahliae Kleb. was examined in the four strawberry breeding lines i.e.'Kent S1', 'Kent o.p.', 'Plena S1', 'Plena o.p'. The strawberry isolate of cv. 'Elsanta' no.1093 of Verticillium dahliae from Pathogen Gene Bank (Poland-Poznań) was used throughout this study. Seedlings of strawberry breeding lines were used for in vitro inoculation at stage of 4 leaves. Their roots were dipped for approximately 1 min in conidial suspensions (inoculum concentration--60 spores at 100x magnification in the microscopic field). Observations of disease symptoms were performed at 15, 30, 45, 60 and 75 days post inoculation. Extent of leaf chlorosis was rated on a scale of 0-4 in which: 0 no symptom. 1 up to 25% chlorotic leaves. 2 up to 50% chlorotic leaves. 3 up to 75% chlorotic leaves. 4 up to 100% chlorotic leaves. Plant response to in vitro inoculation of V. dahliae was different and depended on the breeding line. The most susceptible breeding line was 'Plena S1' and the most resistant was the line 'Kent o.p'. The line 'Kent S1' was more susceptible than the last one, but much more resistant than the line 'Plena o.p'. Seedlings without disease symptoms were observed in all examined lines at 15 and 30 days post inoculation At 45 days post inoculation no plant without disease symptoms was observed. Disease

  8. Entomopathogenic Potential of Verticillium and Acremonium Species (Deuteromycotina: Hyphomycetes)

    DEFF Research Database (Denmark)

    Steenberg, Tove; Humber, Richard A

    1999-01-01

    entomopathogen. The fungi were identified as Verticillium fusisporum, Veritcillium psalliotae, Verticillium lamellicola, and species of Acremonium. Isolates of these fungi were bioassayed against the sweet-potato whitefly (Bemisia tabaci) and against the housefly (Musca domestica) to examine...... their entomopathogenicity. A test was also conducted with a coleopteran (lesser mealworm, Alphitobius diaperinus) to further evaluate the host range for some of the fungi. V. lamellicola was not pathogenic to the two species of insects treated, while varying levels of pathogenicity were shown for the other species....... In general, V. lecanii was the most pathogenic species. Immature whiteflies appeared to be more susceptible to fungal infection than adult houseflies, and the host range for several of the fungi also included lesser mealworm....

  9. Genetics of the partial resistance against race 2 of Verticillium dahliae in lettuce

    Science.gov (United States)

    Lettuce (Lactuca sativa L.) production on the Coastal California is threatened by Verticillium wilt, a soil borne fungal disease caused by Verticillium dahliae that diminishes yield and quality. Two races of V. dahliae were identified on lettuce, race 1 and race 2. Complete resistance to race 1 is c...

  10. Mutational analysis of the Ve1 immune receptor that mediates Verticillium resistance in tomato

    NARCIS (Netherlands)

    Zhang, Z.; Song, Y.; Liu, Chun-Ming; Thomma, B.P.H.J.

    2014-01-01

    Pathogenic Verticillium species are economically important plant pathogens that cause vascular wilt diseases in hundreds of plant species. The Ve1 gene of tomato confers resistance against race 1 strains of Verticillium dahliae and V. albo-atrum. Ve1 encodes an extracellular leucine-rich repeat (eLR

  11. Identification of genetic determinants in potato for resistance to Verticillium wilt

    Science.gov (United States)

    Despite decades of research to control Verticillium wilt (VW), which is caused by the soil-borne fungal pathogens Verticillium dahliae and V. albo-atrum, this disease continues to be a recurrent problem for potato production throughout North America. It can result in yield losses of up to 50% and is...

  12. Breeding and genetics of lettuce for resistance against race 2 Verticillium wilt

    Science.gov (United States)

    Verticillium wilt, caused by Verticillium dahliae Kleb., is an economically important disease of lettuce in central coastal California. Most isolates of the pathogen detected in the Salinas Valley belong to race 1 for which complete resistance exists. However, adequate level of resistance is not ava...

  13. Population biology of Verticillium dahliae isolates from lettuce in the Sallinas Valley of Californis.

    Science.gov (United States)

    Verticillium dahliae is a soil borne fungus and the primary causal agent of Verticillium wilt, which affects many crops worldwide. Many crops grown in the Salinas Valley (SV) of California, including strawberry and lettuce (Lactuca sativa), are susceptible to V. dahliae and severe outbreaks are comm...

  14. Complete Genome Sequence of Phytopathogenic Pectobacterium atrosepticum Bacteriophage Peat1.

    Science.gov (United States)

    Kalischuk, Melanie; Hachey, John; Kawchuk, Lawrence

    2015-08-13

    Pectobacterium atrosepticum is a common phytopathogen causing significant economic losses worldwide. To develop a biocontrol strategy for this blackleg pathogen of solanaceous plants, P. atrosepticum bacteriophage Peat1 was isolated and its genome completely sequenced. Interestingly, morphological and sequence analyses of the 45,633-bp genome revealed that phage Peat1 is a member of the family Podoviridae and most closely resembles the Klebsiella pneumoniae bacteriophage KP34. This is the first published complete genome sequence of a phytopathogenic P. atrosepticum bacteriophage, and details provide important information for the development of biocontrol by advancing our understanding of phage-phytopathogen interactions.

  15. Nachweis von Verticillium dahliae an Pfefferminze (Mentha x piperita L.

    Directory of Open Access Journals (Sweden)

    Gärber, Ute

    2014-09-01

    Full Text Available In Beständen von Pfefferminze wurden Krankheitssymptome beobachtet, die sich in Wuchshemmungen, Stängelverbräunungen, Blattchlorosen und Welke zeigten. Teilweise starben die Pflanzen bzw. einzelne Triebe ab. Als Ursache konnte Verticillium dahliae nachgewiesen werden. In einem Infektionsversuch im Gewächshaus wurden sieben Wochen alte Pfefferminzpflanzen der Sorte Multimentha durch Angießen mit einer Konidiensuspension (106 Konidien/ml Suspension inokuliert. Erste Krankheitssymptome erschienen acht Wochen nach Inokulation. Innerhalb kurzer Zeit nahm der Befall stark zu. Aus den kranken Pflanzenteilen wie Stängel und Blatt konnte der Erreger reisoliert werden.

  16. Maintenance of sex-related genes and the co-occurrence of both mating types in Verticillium dahliae.

    Directory of Open Access Journals (Sweden)

    Dylan P G Short

    Full Text Available Verticillium dahliae is a cosmopolitan, soilborne fungus that causes a significant wilt disease on a wide variety of plant hosts including economically important crops, ornamentals, and timber species. Clonal expansion through asexual reproduction plays a vital role in recurring plant epidemics caused by this pathogen. The recent discovery of recombination between clonal lineages and preliminary investigations of the meiotic gene inventory of V. dahliae suggest that cryptic sex appears to be rare in this species. Here we expanded on previous findings on the sexual nature of V. dahliae. Only 1% of isolates in a global collection of 1120 phytopathogenic V. dahliae isolates contained the MAT1-1 idiomorph, whereas 99% contained MAT1-2. Nine unique multilocus microsatellite types comprised isolates of both mating types, eight of which were collected from the same substrate at the same time. Orthologs of 88 previously characterized sex-related genes from fungal model systems in the Ascoymycota were identified in the genome of V. dahliae, out of 93 genes investigated. Results of RT-PCR experiments using both mating types revealed that 10 arbitrarily chosen sex-related genes, including MAT1-1-1 and MAT1-2-1, were constitutively expressed in V. dahliae cultures grown under laboratory conditions. Ratios of non-synonymous (amino-acid altering to synonymous (silent substitutions in V. dahliae MAT1-1-1 and MAT1-2-1 sequences were indistinguishable from the ratios observed in the MAT genes of sexual fungi in the Pezizomycotina. Patterns consistent with strong purifying selection were also observed in 18 other arbitrarily chosen V. dahliae sex-related genes, relative to the patterns in orthologs from fungi with known sexual stages. This study builds upon recent findings from other laboratories and mounts further evidence for an ancestral or cryptic sexual stage in V. dahliae.

  17. A Glutamic Acid-Rich Protein Identified in Verticillium dahliae from an Insertional Mutagenesis Affects Microsclerotial Formation and Pathogenicity

    Science.gov (United States)

    Li, Guo-Ying; Jia, Pei-Song; Li, Hui; Zhao, Yun-Long; Zhao, Pan; Xia, Gui-Xian; Guo, Hui-Shan

    2010-01-01

    Verticillium dahliae Kleb. is a phytopathogenic fungus that causes wilt disease in a wide range of crops, including cotton. The life cycle of V. dahliae includes three vegetative phases: parasitic, saprophytic and dormant. The dormant microsclerotia are the primary infectious propagules, which germinate when they are stimulated by root exudates. In this study, we report the first application of Agrobacterium tumefaciens-mediated transformation (ATMT) for construction of insertional mutants from a virulent defoliating isolate of V. dahliae (V592). Changes in morphology, especially a lack of melanized microsclerotia or pigmentation traits, were observed in mutants. Together with the established laboratory unimpaired root dip-inoculation approach, we found insertional mutants to be affected in their pathogenicities in cotton. One of the genes tagged in a pathogenicity mutant encoded a glutamic acid-rich protein (VdGARP1), which shared no significant similarity to any known annotated gene. The vdgarp1 mutant showed vigorous mycelium growth with a significant delay in melanized microsclerotial formation. The expression of VdGARP1 in the wild type V529 was organ-specific and differentially regulated by different stress agencies and conditions, in addition to being stimulated by cotton root extract in liquid culture medium. Under extreme infertile nutrient conditions, VdGARP1 was not necessary for melanized microsclerotial formation. Taken together, our data suggest that VdGARP1 plays an important role in sensing infertile nutrient conditions in infected cells to promote a transfer from saprophytic to dormant microsclerotia for long-term survival. Overall, our findings indicate that insertional mutagenesis by ATMT is a valuable tool for the genome-wide analysis of gene function and identification of pathogenicity genes in this important cotton pathogen. PMID:21151869

  18. Assessment of Tolerance Level of some Cotton (Gossypium hirsutum L. Varieties against Verticillium wilt (Verticillium dahliae Kleb. Disease

    Directory of Open Access Journals (Sweden)

    Emine KARADEMIR

    2010-06-01

    Full Text Available The objective of this study was to assess the tolerance level of some cotton varieties against Verticillium wilt (Verticillium dahliae Kleb. disease. Verticillium wilt is one of the major constraint diseases of cotton production worldwide and also in Turkey. The study was carried out at the Southeastern Anatolia Agricultural Research Institute’s naturally infected experimental area during 2004-2006. In this study, 10 different commercial cotton varieties were used as plant material. The experimental design was a randomized complete-block with four replications. During the cotton growing season, foliar disease index (FDI, vascular disease index (VDI and vascular disease rate (VDR were observed in addition to seed cotton yield and some fiber quality characteristics. According to the results, it was determined that with regards to FDI, VDI and VDR, the most tolerant varieties were ‘GW-Teks’, ‘GW-Golda’ and ‘Carmen’, while the most sensitive varieties were ‘Maraş 92’, ‘Sayar 314’ and ‘Stoneville 453’. The other varieties had moderate tolerance levels. The highest seed cotton yield and lint yield were obtained from ‘DP-Deltaopal’ and ‘Stoneville 453’. These results showed that some sensitive varieties had high yield; the reason for this situation may be related with early or late occurrence of the disease. The result of this study indicated that ‘GW-Teks’, ‘GW-Golda’ and ‘Carmen’ varieties must be preferred for infected areas; on the other hand, ‘DP-Deltaopal’ and ‘Stoneville 453’ can be recommended and grown in uninfected areas. Additionally, ‘Carmen’, ‘GW-Teks’ and ‘GW-Golda’ varieties can be used as material for improving disease resistance in cotton breeding programs.

  19. Molecular Cloning and Characterization of Genes Involved in Cotton (Gossypium barbadense L.) Response to Verticillium dahliae

    Institute of Scientific and Technical Information of China (English)

    XU Li; ZHANG Xian-long; ZHU Long-fu; TU Li-li

    2008-01-01

    @@ Verticillium dahliae Kleb.is a necrotrophic plant pathogen which causes serious soil borne vascular disease in cotton.The molecular basis the defense response of cotton to this pathogen is poorly understood.

  20. Transcriptome profiling of Gossypium barbadense inoculated with Verticillium dahliae provides a resource for cotton improvement

    OpenAIRE

    2013-01-01

    Background Verticillium wilt, caused by the fungal pathogen Verticillium dahliae, is the most severe disease in cotton (Gossypium spp.), causing great lint losses worldwide. Disease management could be achieved in the field if genetically improved, resistant plants were used. However, the interaction between V. dahliae and cotton is a complicated process, and its molecular mechanism remains obscure. To understand better the defense response to this pathogen as a means for obtaining more toler...

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

    Science.gov (United States)

    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.

  2. BASIDIOMYCETE-BASED METHOD FOR BIOCONTROL OF PHYTOPATHOGENIC NEMATODES

    Directory of Open Access Journals (Sweden)

    Tiberius BALAEŞ

    2015-12-01

    Full Text Available Phytopathogenic nematodes represent one of the most important groups of pathogens in crops. The use of chemical to control the nematodes attack in crops is decreasing every year due to the concern of the toxicity and side effects of such compounds. In the course for finding alternatives to the use of chemicals, biological control of nematodes is gaining much attention. Some saprotrophic fungi are able to feed on invertebrates, thus becoming efficient agents of control. In this study, three species of basidiomycetes were analyzed for their potential to be used as control agents of phytopathogenic nematodes. Through on in vitro investigation of these potential, one strain – Gymnopilus junonius was further selected for a pot test against Meloidogyne incognita, a very important phytopathogenic species of nematodes. The fungal treatment strongly decreased the M. incognita population on the tested pots, proving the potential of G. junonius strain to be used in biocontrol.

  3. Preservação de fungos fitopatogênicos habitantes do solo Storage of soilborne phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    César J. Bueno

    2006-03-01

    Full Text Available A preservação de fungos fitopatogênicos por longos períodos de tempo é importante para que pesquisas possam ser realizadas a qualquer momento. Os fungos habitantes do solo são organismos que podem produzir estruturas de resistência em face de situações adversas, tais como ausência de hospedeiros e ou condições climáticas desfavoráveis para a sua sobrevivência. O objetivo deste trabalho foi desenvolver metodologias de preservação de estruturas de resistência para os fungos Fusarium oxysporum f.sp. lycopersici raça 2, Macrophomina phaseolina, Rhizoctonia solani AG4 HGI, Sclerotium rolfsii, Sclerotinia sclerotiorum e Verticillium dahliae. O delineamento foi inteiramente casualizado, com um método de produção de estruturas para cada fungo, submetido a três tratamentos [temperatura ambiente de laboratório (28±2ºC, de geladeira (5ºC e de freezer (-20ºC] e com dois frascos por temperatura. Mensalmente, e por um período de um ano, a sobrevivência e o vigor das colônias de cada patógeno foram avaliadas em meios de cultura específicos. Testes de patogenicidade foram realizados após um ano de preservação, com as estruturas que sobreviveram aos melhores tratamentos (temperatura para todos os fungos. As melhores temperaturas (tratamentos para preservar os fungos foram: a F. oxysporum f.sp. lycopersici em temperatura de refrigeração e de freezer (5,2 e 2,9 x 10³ufc.g-1 de talco, respectivamente; b M. phaseolina em temperatura de refrigeração [100% de sobrevivência (S e índice 3 de vigor (V] e S. rolfsii em temperatura ambiente (74,4% S e 1 V e c S. sclerotiorum e V. dahliae, ambos em temperatura de freezer (100% S e 3 V. Após um ano de preservação, somente V. dahliae perdeu a patogenicidade na metodologia desenvolvida.Preservation of soilborne phytopathogenic fungi for long periods of time is important so that researches can be followed up at any moment. Soilborne phytopathogenic fungi are organisms that can

  4. Identification of lettuce genes differentially expressed in a Verticillium dahliae-lettuce interaction by suppression subtractive hybridization

    Science.gov (United States)

    Verticillium wilt, caused by the fungus Verticillium dahliae, is an emerging threat to the U.S. lettuce industry. Lettuce germplasm with resistance to race 1of V. dahliae is available for breeding programs, although germplasm with resistance to race 2 of the pathogen has not been identified. The obj...

  5. Genetics of resistance in lettuce to races 1 and 2 of Verticillium dahliae from different host species

    Science.gov (United States)

    Race 1 resistance against Verticillium dahliae in lettuce was originally shown in the cultivar La Brillante to be conditioned by a single dominant gene (Verticillium resistance 1, Vr1). Multiple, morphologically diverse sources of germplasm have been identified as resistant to race 1. In this study...

  6. Fifteen years of verticillium wilt of lettuce in america’s salad bowl: a tale of immigration, subjugation and abatement.

    Science.gov (United States)

    Lettuce is a popular leafy vegetable that is globally cultivated. The US ranks second in production, with coastal California producing half of the US supply. In 1995, Verticillium wilt caused by the soil borne fungus Verticillium dahliae was identified as a disease of lettuce in coastal California, ...

  7. Genetic dissection of tetraploid cotton resistant to Verticillium wilt using interspecific chromosome segment introgression lines

    Institute of Scientific and Technical Information of China (English)

    Peng; Wang; Zhiyuan; Ning; Ling; Lin; Hong; Chen; Hongxian; Mei; Jun; Zhao; Bingliang; Liu; Xin; Zhang; Wangzhen; Guo; Tianzhen; Zhang

    2014-01-01

    Verticillium wilt(caused by the pathogen Verticillium dahliae) is of high concern for cotton producers and consumers. The major strategy for controlling this disease is the development of resistant cotton(Gossypium spp.) cultivars. We used interspecific chromosome segment introgression lines(CSILs) to identify quantitative trait loci(QTL) associated with resistance to Verticillium wilt in cotton grown in greenhouse and inoculated with three defoliating V. dahliae isolates. A total of 42 QTL, including 23 with resistance-increasing and 19 with resistancedecreasing, influenced host resistance against the three isolates. These QTL were identified and mapped on 18 chromosomes(chromosomes A1, A3, A4, A5, A7, A8, A9, A12, A13, D1, D2,D3, D4, D5, D7, D8, D11, and D12), with LOD values ranging from 3.00 to 9.29. Among the positive QTL with resistance-increasing effect, 21 conferred resistance to only one V. dahliae isolate, suggesting that resistance to V. dahliae conferred by most QTL is pathogen isolate-specific. The At subgenome of cotton had greater effect on resistance to Verticillium wilt than the Dt subgenome. We conclude that pyramiding different resistant QTL could be used to breed cotton cultivars with broad-spectrum resistance to Verticillium wilt.

  8. Cotton GhBAK1 Mediates Verticillium Wilt Resistance and Cell Death

    Institute of Scientific and Technical Information of China (English)

    Xiquan Gao; Fangjun Li; Maoying Li; Ali S.Kianinejad; Jane K.Dever; Terry A.Wheeler; Zhaohu LP

    2013-01-01

    Virus-induced gene silencing (VIGS) offers a powerful approach for functional analysis of individual genes by knocking down their expression.We have adopted this approach to dissect gene functions in cotton resistant to Verticillium wilt,one of the most devastating diseases worldwide.We showed hera that highly efficient VIGS was obtained in a cotton breeding line (CA4002) with partial resistance to Verticillium wilt,and GhMKK2 and Gh Ve 1 are required for its resistance to Verticillium wilt.Arabidopsis AtBAK1/SERK3,a central regulator in plant disease resistance,belongs to a subfamily of somatic embryogenesis receptor kinases (SERKs) with five members,AtSERK1 to AtSERK5.Two BAK1 orthologs and one SERK1 ortholog were identified in the cotton genome.Importantly,GhBAK1 is required for CA4002 resistance to Verticillium wilt.Surprisingly,silencing of GhBAK1 is sufficient to trigger cell death accompanied with production of reactive oxygen species in cotton.This result is distinct from Arabidopsis in which AtBAK1 and AtSERK4 play redundant functions in cell death control.Apparently,cotton has only evolved SERK1 and BAK1 whereas AtSERK4/5 are newly evolved genes in Arabidopsis.Our studies indicate the functional importance of BAK1 in Verticillium wilt resistance and suggest the dynamic evolution of SERK family members in different plant species.

  9. The effects of cotton root exudates on the growth and development of Verticillium dahliae

    Institute of Scientific and Technical Information of China (English)

    Yuxiang WU; Weiping FANG; Shuijin ZHU; Kuiying JIN; Daofan JI

    2008-01-01

    The effects of upland cotton root exudates on the growth and development of Verticillium dahliae were studied, through the compared analysis of the root exudates components between the resistant and suscept-ive cotton materials, using a pair of resistant and sus-ceptive isogenic lines to Verticillium wilt, Z5629 and Z421, as well as 4 other upland cotton cultivars with different resistant levels of Verticillium wilt. The results showed that the amino acids in the root exudates of the resistant cultivars were much less than that of the sus-ceptible ones. Compared with the susceptible ones, there were a lack of aspartic acid, threonine, glutamic acid, alanine, isoleucine, leucine, phenylalanine, lysine and proline in the root exudates from the resistant cul-tivars. On the contrary, arginine was lacking in the sus-ceptive cultivars. The saccharide types in the root exudates were no different between the two kinds of cultivars, but the contents of glucose, fructose and suc-rose in the root exudates of the susceptible varieties were much higher than those in the resistant ones. The experiment of Verticillium dahliae culture showed that the cotton root exudates from resistant cultivars can effectively restrain the spore germination and mycelium growth of Verticillium dahliae, and the argi-nine was the leading amino acid in this inhibitory action, besides the nutrition of the root exudates. However, the cotton root exudates from the susceptive cotton cultivars can improve the growth and develop-ment of Verticillium dahliae effectively; among the amino acid in the exudates, alanine was the most active one in this stimulating function.

  10. AAC as a Potential Target Gene to Control Verticillium dahliae

    Directory of Open Access Journals (Sweden)

    Xiaofeng Su

    2017-01-01

    Full Text Available Verticillium dahliae invades the roots of host plants and causes vascular wilt, which seriously diminishes the yield of cotton and other important crops. The protein AAC (ADP, ATP carrier is responsible for transferring ATP from the mitochondria into the cytoplasm. When V. dahliae protoplasts were transformed with short interfering RNAs (siRNAs targeting the VdAAC gene, fungal growth and sporulation were significantly inhibited. To further confirm a role for VdAAC in fungal development, we generated knockout mutants (ΔVdACC. Compared with wild-type V. dahliae (Vd wt, ΔVdAAC was impaired in germination and virulence; these impairments were rescued in the complementary strains (ΔVdAAC-C. Moreover, when an RNAi construct of VdAAC under the control of the 35S promoter was used to transform Nicotiana benthamiana, the expression of VdAAC was downregulated in the transgenic seedlings, and they had elevated resistance against V. dahliae. The results of this study suggest that VdAAC contributes to fungal development, virulence and is a promising candidate gene to control V. dahliae. In addition, RNAi is a highly efficient way to silence fungal genes and provides a novel strategy to improve disease resistance in plants.

  11. ANTAGONISMO IN VITRO DE Trichoderma spp. A Verticillium dahliae KLEB

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    MARTINS-CORDER M.P.

    1998-01-01

    Full Text Available O presente trabalho visou testar e selecionar isolados selvagens de Trichoderma spp. quanto à capacidade antagônica ao fungo fitopatogênico Verticillium dahliae. Os ensaios in vitro consistiram de testes para avaliar a capacidade hiperparasítica e de antibiose através da produção de metabólitos voláteis e não voláteis. Pela técnica de culturas pareadas, 47 isolados de diversas espécies de Trichoderma foram avaliados e, pelos resultados obtidos, 20 foram selecionados. Através do teste de antibiose, selecionaram-se 7 isolados: T15P e Tal-1 (T. viride, TW6 e CNP311A (T. koningii, CNP17 e TCII (T. harzianum e Tal-10 (T. aureoviride os quais inibiram completamente o crescimento micelial de V. dahliae através da produção de metabólitos. Observações microscópicas demonstraram interações de hifas entre Trichoderma sp. e V. dahliae, tais como: enrolamento, crescimento paralelo de ambos, formação de ganchos.

  12. Combined effects of arthropod herbivores and phytopathogens on plant performance

    DEFF Research Database (Denmark)

    Hauser, Thure Pavlo; Christensen, Stina; Heimes, Christine

    2013-01-01

    1. Many plants are simultaneously attacked by arthropod herbivores and phytopathogens. These may affect each other directly and indirectly, enhancing or reducing the amount of plant resources they each consume. Ultimately, this may reduce or enhance plant performance relative to what should be ex....... However, as interactive impacts also differed among environments and parasite manipulation methods, this suggests that the ability of plants to compensate such losses may depend on environmental conditions and probably also overall infection load.......1. Many plants are simultaneously attacked by arthropod herbivores and phytopathogens. These may affect each other directly and indirectly, enhancing or reducing the amount of plant resources they each consume. Ultimately, this may reduce or enhance plant performance relative to what should...... be expected from the added impacts of herbivore and pathogen when they attack alone. 2. Previous studies have suggested synergistic and antagonistic impacts on plant performance from certain combinations of arthropods and pathogens, for example, synergistic impacts from necrotrophic pathogens together...

  13. Identification of molecular markers associated with Verticillium wilt resistance in alfalfa (Medicago sativa L.) using high-resolution melting.

    Science.gov (United States)

    Zhang, Tiejun; Yu, Long-Xi; McCord, Per; Miller, David; Bhamidimarri, Suresh; Johnson, David; Monteros, Maria J; Ho, Julie; Reisen, Peter; Samac, Deborah A

    2014-01-01

    Verticillium wilt, caused by the soilborne fungus, Verticillium alfalfae, is one of the most serious diseases of alfalfa (Medicago sativa L.) worldwide. To identify loci associated with resistance to Verticillium wilt, a bulk segregant analysis was conducted in susceptible or resistant pools constructed from 13 synthetic alfalfa populations, followed by association mapping in two F1 populations consisted of 352 individuals. Simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers were used for genotyping. Phenotyping was done by manual inoculation of the pathogen to replicated cloned plants of each individual and disease severity was scored using a standard scale. Marker-trait association was analyzed by TASSEL. Seventeen SNP markers significantly associated with Verticillium wilt resistance were identified and they were located on chromosomes 1, 2, 4, 7 and 8. SNP markers identified on chromosomes 2, 4 and 7 co-locate with regions of Verticillium wilt resistance loci reported in M. truncatula. Additional markers identified on chromosomes 1 and 8 located the regions where no Verticillium resistance locus has been reported. This study highlights the value of SNP genotyping by high resolution melting to identify the disease resistance loci in tetraploid alfalfa. With further validation, the markers identified in this study could be used for improving resistance to Verticillium wilt in alfalfa breeding programs.

  14. Identification of molecular markers associated with Verticillium wilt resistance in alfalfa (Medicago sativa L. using high-resolution melting.

    Directory of Open Access Journals (Sweden)

    Tiejun Zhang

    Full Text Available Verticillium wilt, caused by the soilborne fungus, Verticillium alfalfae, is one of the most serious diseases of alfalfa (Medicago sativa L. worldwide. To identify loci associated with resistance to Verticillium wilt, a bulk segregant analysis was conducted in susceptible or resistant pools constructed from 13 synthetic alfalfa populations, followed by association mapping in two F1 populations consisted of 352 individuals. Simple sequence repeat (SSR and single nucleotide polymorphism (SNP markers were used for genotyping. Phenotyping was done by manual inoculation of the pathogen to replicated cloned plants of each individual and disease severity was scored using a standard scale. Marker-trait association was analyzed by TASSEL. Seventeen SNP markers significantly associated with Verticillium wilt resistance were identified and they were located on chromosomes 1, 2, 4, 7 and 8. SNP markers identified on chromosomes 2, 4 and 7 co-locate with regions of Verticillium wilt resistance loci reported in M. truncatula. Additional markers identified on chromosomes 1 and 8 located the regions where no Verticillium resistance locus has been reported. This study highlights the value of SNP genotyping by high resolution melting to identify the disease resistance loci in tetraploid alfalfa. With further validation, the markers identified in this study could be used for improving resistance to Verticillium wilt in alfalfa breeding programs.

  15. Bioprospection of yeasts as biocontrol agents against phytopathogenic molds

    Directory of Open Access Journals (Sweden)

    Márcia Maria Rosa-Magri

    2011-02-01

    Full Text Available Yeasts isolated from sugar cane and maize rhizosphere, leaves and stalks were screened against the phytopathogenic molds Colletotrichum sublineolum and Colletotrichum graminicola, both causal agents of the anthracnose disease in sorghum and maize, respectively. Strains identified as Torulaspora globosa and Candida intermedia were able to inhibit the mold growth, with the first species also exhibiting killer activity. No previous report on the application and potentiality of these yeasts as biocontrol agents were found neither the killer phenotype in Torulaspora globosa.

  16. Bioprospection of yeasts as biocontrol agents against phytopathogenic molds

    OpenAIRE

    Márcia Maria Rosa-Magri; Sâmia Maria Tauk-Tornisielo; Sandra Regina Ceccato-Antonini

    2011-01-01

    Yeasts isolated from sugar cane and maize rhizosphere, leaves and stalks were screened against the phytopathogenic molds Colletotrichum sublineolum and Colletotrichum graminicola, both causal agents of the anthracnose disease in sorghum and maize, respectively. Strains identified as Torulaspora globosa and Candida intermedia were able to inhibit the mold growth, with the first species also exhibiting killer activity. No previous report on the application and potentiality of these yeasts as bi...

  17. Complete Genome Sequence of Phytopathogenic Pectobacterium atrosepticum Bacteriophage Peat1

    OpenAIRE

    Kalischuk, Melanie; Hachey, John; Kawchuk, Lawrence

    2015-01-01

    Pectobacterium atrosepticum is a common phytopathogen causing significant economic losses worldwide. To develop a biocontrol strategy for this blackleg pathogen of solanaceous plants, P. atrosepticum bacteriophage Peat1 was isolated and its genome completely sequenced. Interestingly, morphological and sequence analyses of the 45,633-bp genome revealed that phage Peat1 is a member of the family Podoviridae and most closely resembles the Klebsiella pneumoniae bacteriophage KP34. This is the fir...

  18. [BACILLUS STRAINS'S SCREENING--ACTIVE ANTAGONISTS OF BACTERIAL AND FUNGAL PHYTOPATHOGENS].

    Science.gov (United States)

    Grabova, A Yu; Dragovoz, I V; Kruchkova, L A; Pasichnik, L A; Avdeeva, L V

    2015-01-01

    Antagonistic activity 100 strains of Bacillus bacteria towards to museum and actual strains of phytopathogenic bacteria and fungy was defined. Relation between level of antagonistic activity to phytopathogenic bacteria and genus accessory of the last was shown. The medium level of antagonism to fungal phytopathogens at 30% of the studied strains of Bacillus bacteria was shown. 5 strains of Bacillus sp. with high and medium levels of antagonism to phytopathogens bacterial and fungy nature was selected and considered as perspective for creation of biological preparations for plant protection.

  19. The inheritance of resistance to Verticillium wilt caused by race 1 isolates of Verticillium dahliae in the lettuce cultivar La Brillante.

    Science.gov (United States)

    Hayes, Ryan J; McHale, Leah K; Vallad, Gary E; Truco, Maria Jose; Michelmore, Richard W; Klosterman, Steve J; Maruthachalam, Karunakaran; Subbarao, Krishna V

    2011-08-01

    Verticillium wilt of lettuce caused by Verticillium dahliae can cause severe economic damage to lettuce producers. Complete resistance to race 1 isolates is available in Lactuca sativa cultivar (cv.) La Brillante and understanding the genetic basis of this resistance will aid development of new resistant cultivars. F(1) and F(2) families from crosses between La Brillante and three iceberg cultivars as well as a recombinant inbred line population derived from L. sativa cv. Salinas 88 × La Brillante were evaluated for disease incidence and disease severity in replicated greenhouse and field experiments. One hundred and six molecular markers were used to generate a genetic map from Salinas 88 × La Brillante and for detection of quantitative trait loci. Segregation was consistent with a single dominant gene of major effect which we are naming Verticillium resistance 1 (Vr1). The gene described large portions of the phenotypic variance (R(2) = 0.49-0.68) and was mapped to linkage group 9 coincident with an expressed sequence tag marker (QGD8I16.yg.ab1) that has sequence similarity with the Ve gene that confers resistance to V. dahliae race 1 in tomato. The simple inheritance of resistance indicates that breeding procedures designed for single genes will be applicable for developing resistant cultivars. QGD8I16.yg.ab1 is a good candidate for functional analysis and development of markers suitable for marker-assisted selection.

  20. Preventing an increase in Verticillium wilt incidence in spinach seed production

    DEFF Research Database (Denmark)

    Olesen, Merete Halkjær; Deleuran, Lise Christina; Gislum, René;

    2014-01-01

    A semifield assay was conducted from 2009 to 2011 to distinguish between different preventive methods of reducing Verticillium spp. in spinach seed production. The seed treatments for controlling seed infection levels included Thiram, Signum, Trichoderma harzianum, Gliocladium roseum and Natural II...

  1. Verticillium dahliae LysM effectors differentially contribute to virulence on plant hosts

    NARCIS (Netherlands)

    Kombrink, Anja; Rovenich, Hanna; Shi, Xiaoqian; Rojas-Padilla, Eduardo; Berg-Velthuis, van den Grardy; Domazakis, Emmanouil; Jonge, De Ronnie; Valkenburg, Dirk-Jan; Sánchez-Vallet, Andrea; Seidl, Michael F.; Thomma, Bart P.H.J.

    2017-01-01

    Chitin-binding LysM effectors contribute to virulence of various plant pathogenic fungi that are causal agents of foliar diseases. Here, we report on LysM effectors of the soil-borne fungal vascular wilt pathogen Verticillium dahliae. Comparative genomics revealed three core LysM effectors that are

  2. Potato stem cuttings to study Verticillium dahliae infection for resistance breeding and ‘omics’ studies

    Science.gov (United States)

    The use of consistent and effective methods for early discrimination of resistance to pathogens and selection of appropriate times for tissue sampling are important for experiments focused on global gene expression and metabolomics. Assays for resistance to the vascular pathogen Verticillium dahliae...

  3. The Vayg1 gene controls microsclerotia development and melanin production in Verticillium dahliae

    Science.gov (United States)

    The fungus Verticillium dahliae causes plant vascular diseases on economically important crops and ornamental plants worldwide. This fungus produces darkly pigmented resting structures known as microsclerotia, which are able to survive for years in the soil, and represent a one of the defining char...

  4. Verticillium dahliae Infects, Alters Plant Biomass, and Produces Inoculum on Rotation Crops.

    Science.gov (United States)

    Wheeler, D L; Johnson, D A

    2016-06-01

    Verticillium wilt, caused by Verticillium dahliae, reduces yields of potato and mint. Crop rotation is a potential management tactic for Verticillium wilt; however, the wide host range of V. dahliae may limit the effectiveness of this tactic. The hypothesis that rotation crops are infected by V. dahliae inoculum originating from potato and mint was tested by inoculation of mustards, grasses, and Austrian winter pea with eight isolates of V. dahliae. Inoculum density was estimated from plants and soil. Typical wilt symptoms were not observed in any rotation crop but plant biomass of some crops was reduced, not affected, or increased by infection of specific isolates. Each isolate was host-specific and infected a subset of the rotation crops tested but microsclerotia from at least one isolate were observed on each rotation crop. Some isolates were host-adapted and differentially altered plant biomass or produced differential amounts of inoculum on rotation crops like arugula and Austrian winter pea, which supported more inoculum of specific isolates than potato. Evidence of asymptomatic and symptomatic infection and differential inoculum formation of V. dahliae on rotation crops presented here will be useful in designing rotations for management of Verticillium wilt.

  5. Optimization of Verticillium lecanii spore production in solid-state fermentation on sugarcane bagasse

    NARCIS (Netherlands)

    Shi, Y.; Xu, X.; Zhu, Y.

    2009-01-01

    Verticillium lecanii is an entomopathogen with high potential in biological control of pests. We developed a solid-state fermentation with sugarcane bagasse as carrier absorbing liquid medium to propagate V. lecanii spores. Using statistical experimental design, we optimized the medium composition f

  6. Efficacy of resistance selection to Verticillium wilt in strawberry (Fragaria x ananassa Duch. tissue culture

    Directory of Open Access Journals (Sweden)

    Jadwiga I. Żebrowska

    2012-12-01

    Full Text Available The soil-borne pathogenic fungus Verticillium dahliae Kleb. causes economic losses in crops in temperate regions of the world and hence is the most studied species. Strawberry (Fragaria x ananassa Duch. belongs to plant species susceptible to Verticillium dahliae, although the response to infection caused by this pathogen is varied and depends on the cultivar. Due to a lack of efficient methods in Verticillium wilt elimination, the selection of genetically resistant plant material is a priority direction in breeding programs. Efficacy of resistance selection to Verticillium dahliae Kleb. in strawberry tissue culture was examined on the basis of response to in vitro infection by this pathogenic fungus in two tissue cultured strawberry cultivars, i.e. 'Filon' and 'Teresa'. Culture was conducted for 16 months in an environmentally controlled growth room at 18-20°C, 60-70% relative humidity and light intensity of 100 µm E × m-2 × s-1 on a 16h light / 8h dark cycle. Subcultures were proliferated every 6 weeks on modified Murashige and Skoog medium. Four hundred microplants from each tissue cultured cultivar were inoculated under in vitro conditions at the 4-leaf stage with a homogenate of liquid mycelium of Verticillium dahliae serving as the selecting agent. Disease symptoms were observed at 15, 30, 45, 60, and 75th days post inoculation. The extent of leaf chlorosis was rated on a scale of 0-4. At day 75th post inoculation, the percentage of totally chlorotic plants in micropropagated cv. Teresa reached the value of 76.27%, whereas the proportion of such plants in inoculated tissue cultured cv. Filon reached the value of 89.40%. Also, the index of infection calculated for very severe disease symptoms in the subclone 'Teresa' reached the mean value lower when compared with that calculated for subclone 'Filon' (0.0962 and 0.1150, respectively. These results suggested that the micropropagated cv. Teresa exhibited higher genetic resistance to the

  7. SSH reveals a linkage between a senescence-associated protease and Verticillium wilt symptom development in lettuce (Lactuca sativa)

    Science.gov (United States)

    Suppression subtractive hybridization (SSH) was employed to identify lettuce (Lactuca sativa) genes that are differentially expressed in symptomatic leaves infected with Verticillium dahliae. Genes expressed only in symptomatic leaves included those with homology to pathogenesis-related (PR) protei...

  8. Comparison of Crop Rotation for Verticillium Wilt Management and Effect on Pythium Species in Conventional and Organic Strawberry Production

    OpenAIRE

    Subbarao, Krishna V.

    2009-01-01

    The effects of broccoli and lettuce rotations on population densities of Verticillium dahliae and Pythium spp. in soil and on strawberry (Fragaria × ananassa) growth, yield, and Verticillium wilt were evaluated in conventional and organic production systems in California for 2 years. Under both management systems, strawberry was planted after two successive crops of broccoli or lettuce. The control treatment in the conventional field was strawberry planted in soils fumigated with methyl bromi...

  9. Extracellular Membrane Vesicles and Phytopathogenicity of Acholeplasma laidlawii PG8

    Directory of Open Access Journals (Sweden)

    Vladislav M. Chernov

    2012-01-01

    Full Text Available For the first time, the phytopathogenicity of extracellular vesicles of Acholeplasma laidlawii PG8 (a ubiquitous mycoplasma that is one of the five common species of cell culture contaminants and is a causative agent for phytomycoplasmoses in Oryza sativa L. plants was studied. Data on the ability of extracellular vesicles of Acholeplasma laidlawii PG8 to penetrate from the nutrient medium into overground parts of Oryza sativa L. through the root system and to cause alterations in ultrastructural organization of the plants were presented. As a result of the analysis of ultrathin leaf sections of plants grown in medium with A. laidlawii PG8 vesicles, we detected significant changes in tissue ultrastructure characteristic to oxidative stress in plants as well as their cultivation along with bacterial cells. The presence of nucleotide sequences of some mycoplasma genes within extracellular vesicles of Acholeplasma laidlawii PG8 allowed a possibility to use PCR (with the following sequencing to perform differential detection of cells and bacterial vesicles in samples under study. The obtained data may suggest the ability of extracellular vesicles of the mycoplasma to display in plants the features of infection from the viewpoint of virulence criteria—invasivity, infectivity—and toxigenicity—and to favor to bacterial phytopathogenicity.

  10. Mapping QTL associated with Verticillium dahliae resistance in the cultivated strawberry (Fragaria × ananassa).

    Science.gov (United States)

    Antanaviciute, L; Šurbanovski, N; Harrison, N; McLeary, K J; Simpson, D W; Wilson, F; Sargent, D J; Harrison, R J

    2015-01-01

    A biparental cross of octoploid strawberry segregating for resistance to Verticillium dahliae, the causative agent of Verticillium wilt, was screened under field conditions for three seasons. Average wilt scores were significantly associated with multiple QTL, which were mostly significant across all years. Markers significantly associated with the traits were used to screen material with known wilt resistance and susceptibility phenotypes. A clear and statistically significant relationship was observed between resistant, tolerant and susceptible material and the total number of markers present in the different resistance classes. In field situations resistance QTL appear to behave in an additive manner. These markers are abundant in the cultivated strawberry germplasm indicating that, despite the large number of markers, clear genetic gain is possible through marker-assisted breeding.

  11. Biocontrol potential of Trichoderma harzianum in controlling wilt disease of pistachio caused by Verticillium dahliae

    Directory of Open Access Journals (Sweden)

    Fotoohiyan Zeinab

    2017-06-01

    Full Text Available Verticillium wilt caused by Verticillium dahliae, is one of the most devastating diseases in pistachio orchards in the world including Iran. In search for an effective non-chemical strategy for the management of this disease, we evaluated the biocontrol potential of Trichoderma harzianum isolates obtained from the rhizosphere of healthy pistachio trees in different locations of the Kerman province of Iran against V. dahliae under laboratory and greenhouse conditions. Dual culture tests in the laboratory were conducted in a completely randomized design using 72 T. harzianum isolates. Twenty isolates showed the highest in vitro antagonistic activity. The results indicated that all 20 isolates were capable of inhibiting the mycelial growth of V. dahliae significantly. Among them, isolates Tr8 and Tr19 were the most effective by 88.89% and 85.12% inhibition, respectively. Extracted cell free metabolites of all effective isolates also inhibited the growth of V. dahliae in the culture medium significantly. According to the results, isolates Tr4 and Tr6 inhibited fungal pathogen growth by 94.94% and 88.15% respectively, through production of non-volatile metabolites. In the evaluation of volatile metabolites, isolates Tr5 and Tr4 were the most effective by 26.27% and 24.49% growth inhibition, respectively. Based on the results of the in vitro experiments, the five most effective isolates were selected for evaluation under greenhouse conditions for their biocontrol potential in controlling Verticillium wilt of pistachio. Results of the greenhouse, (in vivo experiments were positive and indicated that the occurrence of wilt disease in plants treated with the antagonists alone or in combination with pathogenic fungus was lower than in plants inoculated with pathogen alone. The overall results of this study suggest that Trichoderma fungal antagonist may be an effective biocontrol agent for the control of Verticillium wilt of pistachio.

  12. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7

    NARCIS (Netherlands)

    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 PIC

  13. Phytopathogenic micromycetes in central Poland. I. Peronosporales and Erysiphales

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    Małgorzata Ruszkiewicz-Michalska

    2014-08-01

    Full Text Available The present paper begins a new series of studies investigating the occurrence of phytopathogenic micromycetes in Central Poland. Fungi of the orders Peronosporales and Erysiphalas are discussed in part one. Relevant knowledge on the subject is surveyed, and a list of published records (46 taxa as well as the findings collected by the present authors (99 taxa is provided. The list comprises 2 species new for biota of Poland - Microsphaera deutziae Bunkina and Microsphaera elevata Burrill, 10 rare and many common fungal species that had not been previously re00rded in this area as well as 21 plant taxa, mostly species of deliberate or accidental anthropogenic origin, that are new hosts of the parasites formerly listed in Poland.

  14. Growth rate inhibition of phytopathogenic fungi by characterized chitosans

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    Enio N. Oliveira Junior

    2012-06-01

    Full Text Available The inhibitory effects of fifteen chitosans with different degrees of polymerization (DP and different degrees of acetylation (F A on the growth rates (GR of four phytopathogenic fungi (Alternaria alternata, Botrytis cinerea, Penicillium expansum, and Rhizopus stolonifer were examined using a 96-well microtiter plate and a microplate reader. The minimum inhibitory concentrations (MICs of the chitosans ranged from 100 µg × mL-1 to 1,000 µg × mL-1 depending on the fungus tested and the DP and F A of the chitosan. The antifungal activity of the chitosans increased with decreasing F A. Chitosans with low F A and high DP showed the highest inhibitory activity against all four fungi. P. expansum and B. cinerea were relatively less susceptible while A. alternata and R. stolonifer were relatively more sensitive to the chitosan polymers. Scanning electron microscopy of fungi grown on culture media amended with chitosan revealed morphological changes.

  15. Phytopathogen effectors subverting host immunity: different foes, similar battleground.

    Science.gov (United States)

    Dou, Daolong; Zhou, Jian-Min

    2012-10-18

    Phytopathogenic bacteria, fungi, and oomycetes invade and colonize their host plants through distinct routes. These pathogens secrete diverse groups of effector proteins that aid infection and establishment of different parasitic lifestyles. Despite this diversity, a comparison of different plant-pathogen systems has revealed remarkable similarities in the host immune pathways targeted by effectors from distinct pathogen groups. Immune signaling pathways mediated by pattern recognition receptors, phytohormone homeostasis or signaling, defenses associated with host secretory pathways and pathogen penetrations, and plant cell death represent some of the key processes controlling disease resistance against diverse pathogens. These immune pathways are targeted by effectors that carry a wide range of biochemical functions and are secreted by completely different pathogen groups, suggesting that these pathways are a common battleground encountered by many plant pathogens. Copyright © 2012 Elsevier Inc. All rights reserved.

  16. A unique endoglucanase-encoding gene cloned from the phytopathogenic fungus Macrophomina phaseolina.

    OpenAIRE

    Wang, H; Jones, R W

    1995-01-01

    The deduced amino acid sequence derived from a Macrophomina phaseolina beta-1,4-endoglucanase-encoding gene revealed 48% identity (over 119 amino acids) with egl1 from the phytopathogen Pseudomonas solanacearum. Its similarity to saprophyte endoglucanases was not significant. Its minimum substrate size, unlike that of any known saprophyte endoglucanase, was cellopentaose. The unique characteristics of M. phaseolina egl1-encoded endoglucanase suggest that it is phytopathogen specific.

  17. A unique endoglucanase-encoding gene cloned from the phytopathogenic fungus Macrophomina phaseolina.

    Science.gov (United States)

    Wang, H; Jones, R W

    1995-05-01

    The deduced amino acid sequence derived from a Macrophomina phaseolina beta-1,4-endoglucanase-encoding gene revealed 48% identity (over 119 amino acids) with egl1 from the phytopathogen Pseudomonas solanacearum. Its similarity to saprophyte endoglucanases was not significant. Its minimum substrate size, unlike that of any known saprophyte endoglucanase, was cellopentaose. The unique characteristics of M. phaseolina egl1-encoded endoglucanase suggest that it is phytopathogen specific.

  18. Hyphopodium-Specific VdNoxB/VdPls1-Dependent ROS-Ca2+ Signaling Is Required for Plant Infection by Verticillium dahliae.

    Directory of Open Access Journals (Sweden)

    Yun-Long Zhao

    2016-07-01

    Full Text Available Verticillium dahliae is a phytopathogenic fungus obligate in root infection. A few hyphopodia differentiate from large numbers of hyphae after conidia germination on the root surface for further infection. However, the molecular features and role of hyphopodia in the pathogenicity of V. dahliae remain elusive. In this study, we found that the VdPls1, a tetraspanin, and the VdNoxB, a catalytic subunit of membrane-bound NADPH oxidases for reactive oxygen species (ROS production, were specifically expressed in hyphopodia. VdPls1 and VdNoxB highly co-localize with the plasma membrane at the base of hyphopodia, where ROS and penetration pegs are generated. Mutant strains, VdΔnoxb and VdΔpls1, in which VdPls1 and VdNoxB were deleted, respectively, developed defective hyphpodia incapable of producing ROS and penetration pegs. Defective plasma membrane localization of VdNoxB in VdΔpls1 demonstrates that VdPls1 functions as an adaptor protein for the recruitment and activation of the VdNoxB. Furthermore, in VdΔnoxb and VdΔpls1, tip-high Ca2+ accumulation was impaired in hyphopodia, but not in vegetative hyphal tips. Moreover, nuclear targeting of VdCrz1 and activation of calcineurin-Crz1 signaling upon hyphopodium induction in wild-type V. dahliae was impaired in both knockout mutants, indicating that VdPls1/VdNoxB-dependent ROS was specifically required for tip-high Ca2+ elevation in hyphopodia to activate the transcription factor VdCrz1 in the regulation of penetration peg formation. Together with the loss of virulence of VdΔnoxb and VdΔpls1, which are unable to initiate colonization in cotton plants, our data demonstrate that VdNoxB/VdPls1-mediated ROS production activates VdCrz1 signaling through Ca2+ elevation in hyphopodia, infectious structures of V. dahliae, to regulate penetration peg formation during the initial colonization of cotton roots.

  19. Temperature modulates the secretome of the phytopathogenic fungus Lasiodiplodia theobromae

    Directory of Open Access Journals (Sweden)

    Carina Félix

    2016-08-01

    Full Text Available Environmental alterations modulate host-microorganism interactions. Little is known about how climate changes can trigger pathogenic features on symbiont or mutualistic microorganisms. Current climate models predict increased environmental temperatures.The exposing of phytopathogens to these changing conditions can have particularly relevant consequences for economically important species and for humans. The impact on pathogen/host interaction and the shift on their biogeographical range can induce different levels of virulence in new hosts, allowing massive losses in agricultural and health fields.Lasiodiplodia theobromae is a phytopathogenic fungus responsible for a number of diseases in various plants. It has also been described as an opportunist pathogen in humans, causing infections with different levels of severity. Lasiodiplodia theobromae has a high capacity of adaptation to different environments, such as woody plants, moist argillaceous soils or even humans, being able to grow and infect hosts in a wide range of temperatures (9ºC-39ºC. Nonetheless, the effect of an increase of temperature, as predicted in climate change models, on L. theobromae is unknown.Here we explore the effect of temperature on two strains of L. theobromae – an environmental strain CAA019, and a clinical strain, CBS339.90.We show that both strains are cytotoxic to mammalian cells but while the environmental strain is cytotoxic mainly at 25ºC, the clinical strain is cytotoxic mainly at 30ºC and 37ºC.Extracellular gelatinolytic, xylanolytic, amylolytic and cellulolytic activities at 25ºC and 37ºC were characterized by zymography and the secretome of both strains grown at 25ºC, 30ºC and 37ºC were characterized by electrophoresis and by Orbitrap LC-MS/MS. More than 75% of the proteins were identified, mostly enzymes (glycosyl hydrolases and proteases. The strains showed different protein profiles, which were affected by growth temperature. Also, strain

  20. Arbuscular mycorrhizal fungi (AMF) as bio protector agents against wilt induced by Verticillium spp. in pepper

    Energy Technology Data Exchange (ETDEWEB)

    Goicoechea, N.; Garmendia, I.; Sanchez-Diaz, M.; Aguirreolea, J.

    2010-07-01

    Verticillium dahliae Kleb. is a vascular pathogen that alters water status and growth of pepper plants and causes drastic reductions in yield. Its control is difficult because it can survive in field soil for several years. The application of arbuscular mycorrhizal fungi (AMF) as bio protector agents against V. dahliae is an alternative to the use of chemicals which, in addition, is more respectful with the environment. The establishment of the mutualistic association of plant roots and AMF involves a continuous cellular and molecular dialogue between both symbionts that includes the pre activation of plant defense responses that may enhance the resistance or tolerance of mycorrhizal plants to soil-borne pathogens. Some AMF can improve the resistance of Capsicum annuum L. against V. dahliae. This is especially relevant for pepper cultivars (i.e. cv. Piquillo) that exhibit high susceptibility to this pathogen. Compared with non-mycorrhizal plants, mycorrhizal pepper can exhibit more balanced antioxidant metabolism in leaves along the first month after pathogen inoculation, which may contribute to delay both the development of disease symptoms and the decrease of photosynthesis in Verticillium-inoculated plants with the subsequent benefit for yield. In stems, mycorrhizal pepper show earlier and higher deposition of lignin in xylem vessels than non mycorrhizal plants, even in absence of the pathogen. Moreover, AMF can induce new isoforms of acidic chitinases and superoxide dismutase in roots. Mycorrhizal-specific induction of these enzymatic activities together with enhanced peroxidase and phenylalanine ammonia-lyase in roots may also be involved in the bio protection of Verticillium-induced wilt in pepper by AMF. (Author) 81 refs.

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

    Science.gov (United States)

    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.

  2. Potential of Endophytic Fungi Isolated from Cotton Roots for Biological Control against Verticillium Wilt Disease

    Science.gov (United States)

    Wang, Lingfei; Li, Zhifang; Shi, Yongqiang; Zhao, LiHong; Feng, Zili; Zhu, Heqin

    2017-01-01

    Verticillium wilt is a soil-borne disease, and severely limits the development of cotton production. To investigate the role of endophytic fungi on Verticillium wilt, CEF-818 (Penicillium simplicissimum), CEF-714 (Leptosphaeria sp.), CEF-642 (Talaromyces flavus.) and CEF-193 (Acremonium sp.) isolated from cotton roots were used to assess their effects against cotton wilt disease caused by a defoliating V. dahliae strain Vd080. In the greenhouse, all treatments significantly reduced disease incidence and disease index, with the control efficacy ranging from 26% (CEF-642) to 67% (CEF-818) at 25 days (d) after inoculation. In the disease nursery, compared to controls (with disease incidence of 33.8% and disease index of 31), CEF-818, CEF-193, CEF-714 and CEF-642 provided a protection effect of 69.5%, 69.2%, 54.6% and 45.7%, respectively. Especially, CEF-818 and CEF-714 still provided well protection against Verticillium wilt with 46.9% and 56.6% or 14.3% and 33.7% at the first peak of the disease in heavily infected field, respectively (in early July). These results indicated that these endophytes not only delayed but also reduced wilt symptoms on cotton. In the harvest, the available cotton bolls of plant treated with CEF-818 and CEF-714 increased to 13.1, and 12.2, respectively. And the seed cotton yield significantly increased after seed bacterization with CEF-818 (3442.04 kg/ha) compared to untreated control (3207.51 kg/ha) by 7.3%. Furtherly, CEF-818 and CET-714 treatment increased transcript levels for PAL, PPO, POD, which leads to the increase of cotton defense reactions. Our results indicate that seed treatment of cotton plants with CEF-818 and CET-714 can help in the biocontrol of V. dahliae and improve seed cotton yield in cotton fields. This study provided a better understanding of cotton-endophyte interactions which will aid in developing effective biocontrol agents for Verticillium wilt of cotton in futhre. PMID:28107448

  3. Physiological mechanisms involved in resistance to cotton verticillium wilt induced by AM fungi

    Institute of Scientific and Technical Information of China (English)

    LIU Bing-jiang; LIU Run-jin

    2004-01-01

    @@ It was proved that arbuscular mycorrhizal (AM) fungi played an important role in increasing plant resistance to soilborne pathogens, especially when plants were pre-inoculated with AM fungi.Mechanisms involved in this phenomenon are not yet well understood. On the basis of the former experiment results in our lab, effects of AM fungi on cotton Verticillium wilt and the mechanisms of increasing disease resisitance by the tested fungi were studied in pot culture under greenhouse conditions. Two cotton cutivars Litai 8 and 86-1 which are susceptible to Verticillium dahliae were pre-inoculated with Glomus fasiculatum, and Gigaspora margarita, then inoculated with the strain of Verticillium dahliae, namely "An-Yang" (belong to intermediate virulent type) 30 days after the former inoculation. Results showed that AM fungi could improve the growth and development of cotton plants, increase plants dry mass, decrease incidence and disease index of Verticillium wilt of cotton plants, inhibit the infection and development of V. dahliae to different extent in the rhizosphere of cotton pre-inoculated with AM fungi, while the colonization and spore numbers of AM fungi were not reduced significantly by this pathogen. The defence enzymes, such as phenylalanine ammonia-lyase (PAL), chitinase, β-1, 3-glucanase, peroxidase, polyphenoloxidase (PPO) were induced, and their activities and peak increased by AM fungi in roots and leaves, and the increasing speed and peak of the enzyme activity were higher in treatment with AM fungus preinoculation than the inoculation with only V. dahliae, which suggested that defense response was activated by AM fungi, and then made the cotton to react strongly and rapidly to the infection of V. dahliae. In addition, AM fungi decreased the content of malondiadehyde (MDA) in cotton roots and leaves,protected membrane system and alleviated the damage caused by the pathogen. The AM fungus,Glomus fasiculatum showed the superior effects of biological

  4. Paecilomyces lilacinus and Verticillium chlamydosporium Fungi as Biological Control of Fasciolosis

    Directory of Open Access Journals (Sweden)

    Riza Zainuddin Ahmad

    2013-09-01

    Full Text Available Fasciolosis is a worm disease caused of Fasciola gigantica and an important problem in husbandry especially for cattle. Controlling of this worm disease can be conducted by prevention and treatment. The use of antihelminthic is commonly causes a resistance problem. Natural control by mold such as Paecilomyces lilacinus and Verticillium chlamydosporium can be applied to reduce egg of F. gigantica. Although it was recently found, in vitro study gave satisfied result. This gives a new hope in controlling the disease although the extend application still needs to be studied. This paper discussed about the use of P. lilacinus and V. chlamydosporium for reducing F. gigantica population.

  5. Soil temperature determines the reaction of olive cultivars to Verticillium dahliae pathotypes.

    Directory of Open Access Journals (Sweden)

    Rocío Calderón

    Full Text Available Development of Verticillium wilt in olive, caused by the soil-borne fungus Verticillium dahliae, can be influenced by biotic and environmental factors. In this study we modeled i the combined effects of biotic factors (i.e., pathotype virulence and cultivar susceptibility and abiotic factors (i.e., soil temperature on disease development and ii the relationship between disease severity and several remote sensing parameters and plant stress indicators.Plants of Arbequina and Picual olive cultivars inoculated with isolates of defoliating and non-defoliating V. dahliae pathotypes were grown in soil tanks with a range of soil temperatures from 16 to 32°C. Disease progression was correlated with plant stress parameters (i.e., leaf temperature, steady-state chlorophyll fluorescence, photochemical reflectance index, chlorophyll content, and ethylene production and plant growth-related parameters (i.e., canopy length and dry weight.Disease development in plants infected with the defoliating pathotype was faster and more severe in Picual. Models estimated that infection with the defoliating pathotype was promoted by soil temperatures in a range of 16 to 24°C in cv. Picual and of 20 to 24°C in cv. Arbequina. In the non-defoliating pathotype, soil temperatures ranging from 16 to 20°C were estimated to be most favorable for infection. The relationship between stress-related parameters and disease severity determined by multinomial logistic regression and classification trees was able to detect the effects of V. dahliae infection and colonization on water flow that eventually cause water stress.Chlorophyll content, steady-state chlorophyll fluorescence, and leaf temperature were the best indicators for Verticillium wilt detection at early stages of disease development, while ethylene production and photochemical reflectance index were indicators for disease detection at advanced stages. These results provide a better understanding of the differential

  6. Polyamine Metabolism in Fungi with Emphasis on Phytopathogenic Species

    Directory of Open Access Journals (Sweden)

    Laura Valdés-Santiago

    2012-01-01

    Full Text Available Polyamines are essential metabolites present in all living organisms, and this subject has attracted the attention of researchers worldwide interested in defining their mode of action in the variable cell functions in which they are involved, from growth to development and differentiation. Although the mechanism of polyamine synthesis is almost universal, different biological groups show interesting differences in this aspect that require to be further analyzed. For these studies, fungi represent interesting models because of their characteristics and facility of analysis. During the last decades fungi have contributed to the understanding of polyamine metabolism. The use of specific inhibitors and the isolation of mutants have allowed the manipulation of the pathway providing information on its regulation. During host-fungus interaction polyamine metabolism suffers striking changes in response to infection, which requires examination. Additionally the role of polyamine transporter is getting importance because of its role in polyamine regulation. In this paper we analyze the metabolism of polyamines in fungi, and the difference of this process with other biological groups. Of particular importance is the difference of polyamine biosynthesis between fungi and plants, which makes this process an attractive target for the control of phytopathogenic fungi.

  7. Data integration and knowledge management to facilitate research on plant-pathogen interactions: case study Solanum tuberosum – Verticillium

    Science.gov (United States)

    Verticillium wilt is an important disease causing considerable economic losses in potato production globally. Soil and environmental conditions affect symptom expression and the effect of the disease on yield. A ten year field trial has been conducted in order to better understand the dynamics of wi...

  8. Media for efficient generating nitrate- - nonutilizing (NIT) mutants of Verticillium dahliae, Colletotrichum gloeosporioides, Colletotrichum lindemuthianum and Fusarium oxysporum.

    NARCIS (Netherlands)

    Rataj-Guranowska, M.; Pieczul, K.; Nowak, E.; Hiemstra, J.A.; Drapikowska, M.

    2001-01-01

    The effect of several media amended wit potassium chlorate (1.5% and 6%) on generation of nit mutants, especially nit M mutants from Verticillium dahliae, Colletotrichum gloeosporioides, Colletotrichum lindemuthianum and Fusarium oxysporum were studied. For all species minimal medium with 6% chlorat

  9. Improvement of the efficacy of Verticillium lecanii used in biocontrol of Sphaerotheca fuliginea by addition of oil formulations

    NARCIS (Netherlands)

    Verhaar, M.A.; Hijwegen, T.; Zadoks, J.C.

    1999-01-01

    Biocontrol of cucumber powdery mildew, Sphaerotheca fuliginea, by Verticillium lecanii is seriously hampered at low humidities. The effect is especially marked at low humidity (60% RH) during the three hours following the application of V. lecanii spores suspended in water. Formulations of V.

  10. Media for efficient generating nitrate- - nonutilizing (NIT) mutants of Verticillium dahliae, Colletotrichum gloeosporioides, Colletotrichum lindemuthianum and Fusarium oxysporum.

    NARCIS (Netherlands)

    Rataj-Guranowska, M.; Pieczul, K.; Nowak, E.; Hiemstra, J.A.; Drapikowska, M.

    2001-01-01

    The effect of several media amended wit potassium chlorate (1.5% and 6%) on generation of nit mutants, especially nit M mutants from Verticillium dahliae, Colletotrichum gloeosporioides, Colletotrichum lindemuthianum and Fusarium oxysporum were studied. For all species minimal medium with 6%

  11. Comparative Proteomic Analysis of Gossypium thurberi in Response to Verticillium dahliae Inoculation

    Directory of Open Access Journals (Sweden)

    Weiping Fang

    2015-10-01

    Full Text Available Verticillium wilt is threatening cotton productivity globally. This disease is caused by soil-borne Verticillium dahliae which directly infects cotton roots, and exclusively colonizes and occludes xylem vessels, finally resulting in necrosis, defoliation, and most severely, plant death. For the first time, iTRAQ (isobaric tags for relative and absolute quantification was applied to screen the differentially expressed proteins of Gossypium thurberi inoculated with V. dahliae. A total of 6533 proteins were identified from the roots of G. thurberi after inoculation with V. dahliae, and 396 showed up- and 279 down-regulated in comparison to a mock-inoculated roots. Of these identified proteins, the main functional groups were those involved in cell wall organization and reinforcement, disease-resistant chemicals of secondary metabolism, phytohormone signaling, pathogenesis-related proteins, and disease-resistant proteins. Physiological and biochemical analysis showed that peroxidase activity, which promotes the biosynthesis and accumulation of lignin, was induced early in the hypocotyl after inoculation with V. dahliae. Similarly, salicylic acid also accumulated significantly in hypocotyl of the seedlings after inoculation. These findings provide an important knowledge of the molecular events and regulatory networks occurring during G. thurberi-V. dahliae interaction, which may provide a foundation for breeding disease-resistance in cotton.

  12. Mapping and Quantitative Trait Loci Analysis of Verticillium Wilt Resistance Genes in Cotton

    Institute of Scientific and Technical Information of China (English)

    Hong-Mei Wang; Zhong-Xu Lin; Xian-Long Zhang; Wei Chen; Xiao-Ping Guo; Yi-Chun Nie; Yun-Hai Li

    2008-01-01

    Verticillium wilt is one of the most serious constraints to cotton production in almost all of the cotton-growing countries. In this study, "XinLuZao1" (XLZ1), a susceptible cultivar Gossypium hirsutum L. and "Hai7124" (H7124), a resistant line G. barbadense, and their F2:3 families were used to map and study the disease Index induced by verticillium wilt. A total of 430 SSR loci were mapped into 41 linkage groups; the map spanned 3 745.9 cM and the average distance between adjacent loci was 8.71 cM. Four and five quantitative trait loci (QTLs) were detected based on the disease index investigated on July 22 and August 24 in 2004, respectively. These nine QTLs explained 10.63-28.83% of the phenotypic variance, six of them were located on the D sub-genome. Two QTLs located In the same marker intervals may partly explain the significant correlation of the two traits. QTLs explaining large phenotypic variation were identified in this study, which may be quite useful in cotton anti-disease breeding.

  13. Effects of methyl jasmonate and gums formed in stone fruit trees on in vitro ethylene production by mycelium of verticillium dahliae and alternaria alternata

    National Research Council Canada - National Science Library

    Elżbieta Węgrzynowicz-Lesiak; Anna Jarecka Boncela; Justyna Góraj; Marian Saniewski

    2013-01-01

    .... In this study, we describe the effect of methyl jasmonate (JA-Me) and gums formed in stone fruit trees on the growth and in vitro ethylene production by mycelium of Verticillium dahliae and Alternaria alternata...

  14. Draft Genome Sequence of the Rhizobacterium Pseudomonas chlororaphis PCL1601, Displaying Biocontrol against Soilborne Phytopathogens.

    Science.gov (United States)

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

    2017-04-06

    In this study, we present the draft genome sequence of the bacterial strain Pseudomonas chlororaphis PCL1601. This bacterium was isolated from the rhizosphere of healthy avocado trees and displayed antagonistic and biological control activities against different soilborne phytopathogenic fungi and oomycetes. Copyright © 2017 Vida et al.

  15. Fungicide sensitivity of selected Verticillium fungicola isolates from Agaricus bisporus farms

    Directory of Open Access Journals (Sweden)

    Potočnik Ivana

    2008-01-01

    Full Text Available Five isolates of Verticillium fungicola, isolated from diseased fruiting bodies of Agaricus bisporus collected from mushroom farms in Serbia during 2002-2003, were studied. By observing their colony morphology under different growth conditions and their pathogenic characteristics, the isolates were identified as V. fungicola var. fungicola. The peat/lime casing was the primary source of infection. Testing of sensitivity to selected fungicides showed that all isolates were highly resistant to benomyl (EC50 values were higher than 200.00 mg/l, moderately sensitive to iprodione (EC50 values were between 11.93 and 22.80 mg/l, and highly sensitive to prochloraz-Mn (EC50 values were less than 3.00 mg/l.

  16. THE INSECT PATHOGENIC FUNGUS Verticillium lecanii (Zimm. Viegas AND ITS USE FOR PESTS CONTROL: A REVIEW

    Directory of Open Access Journals (Sweden)

    Thiery B C ALAVO

    2015-08-01

    Full Text Available Chemical insecticides play an important role in the control of plant damage and plant diseases. However, extensive use of these products has led to the disruption of ecosystems because of several reasons such as death of non-target species, accumulation of pesticide residues in the environment and food, and buildup of pesticide resistance in the target species. Biological control is one of the alternatives to chemical pesticides and it can be described as the limitation of the abundance of living organisms and their products by other living organisms. Predators, parasitoids, fungi and other beneficial organisms can be used for the biocontrol of insect pests. The fungus Verticillium lecanii is one of the members of Deuteromycetes and it can be used for crop protection. This paper is a review of the international literature related to V. lecanii for the bio-control of insects of agricultural importance.

  17. Development of an assay for rapid detection and quantification of Verticillium dahliae in soil.

    Science.gov (United States)

    Bilodeau, Guillaume J; Koike, Steven T; Uribe, Pedro; Martin, Frank N

    2012-03-01

    ABSTRACT Verticillium dahliae is responsible for Verticillium wilt on a wide range of hosts, including strawberry, on which low soil inoculum densities can cause significant crop loss. Determination of inoculum density is currently done by soil plating but this can take 6 to 8 weeks to complete and delay the grower's ability to make planting decisions. To provide a faster means for estimating pathogen populations in the soil, a multiplexed TaqMan real-time polymerase chain reaction (PCR) assay based on the ribosomal DNA (rDNA) intergenic spacer (IGS) was developed for V. dahliae. The assay was specific for V. dahliae and included an internal control for evaluation of inhibition due to the presence of PCR inhibitors in DNA extracted from soil samples. An excellent correlation was observed in regression analysis (R(2) = 0.96) between real-time PCR results and inoculum densities determined by soil plating in a range of field soils with pathogen densities as low as 1 to 2 microsclerotia/g of soil. Variation in copy number of the rDNA was also evaluated among isolates by SYBR Green real-time PCR amplification of the V. dahliae-specific amplicon compared with amplification of several single-copy genes and was estimated to range from ≈24 to 73 copies per haploid genome, which translated into possible differences in results among isolates of ≈1.8 cycle thresholds. Analysis of the variation in results of V. dahliae quantification among extractions of the same soil sample indicated that assaying four replicate DNA extractions for each field sample would provide accurate results. A TaqMan assay also was developed to help identify colonies of V. tricorpus on soil plates.

  18. Evidences for involvement of endogenous cAMP in Arabidopsis defense responses to Verticillium toxins

    Institute of Scientific and Technical Information of China (English)

    Jing JIANG; Ling Wen FAN; Wei Hua WU

    2005-01-01

    Although there were reports suggesting the involvement of endogenous cAMP in plant defense signaling cascades,there is no direct evidence supporting this notion yet and the detailed mechanism is unclear. In the present study, we have used pathogenic fungi Verticillium dahliae and Arabidopsis plants as a model system of plant-microb interaction to demonstrate the function of endogenous cAMP in Arabidopsis defense responses. Both V. dahliae inoculation and Verticillium toxins injection induced typical "wilt" symptoms in Arabidopsis seedlings. When either 8-Br-AMP (a membrane permeable cAMP analogue) or salicylic acid (SA) was applied to Arabidopsis, the plants became resistant to V. dahliae toxins. However, addition of 8-Br-AMP did not increase the resistance of Arabidopsis transgenic plants deficient in SA to the toxins, suggesting that cAMP might act upstream of SA in plant defense signaling pathway.Indeed, 8-Br-cAMP and forskolin, an activator of adenylyl cyclase, significantly stimulated the endogenous SA level in plants, whereas DDA, an inhibitor of adenylyl cyclase dramatically reduced toxin-induced SA increase. Both the endogenous cAMP and SA increased significantly in Arabidopsis seedlings treated with toxins. Furthermore, transcription level of pathogenesis-related protein 1 gene (PR1) was strongly induced by both 8-Br-cAMP and the toxin treatment. Taken together, our data demonstrate that endogenous cAMP is involved in plant defense responses against Verticilliumsecreted toxins by regulating the production of the known signal SA in plant defense pathway.

  19. Isolation and characterization of a chitinase gene from entomopathogenic fungus Verticillium lecanii Isolamento e caracterização de um gene de quitinase do fungo entomopatogênico Verticillium lecanii

    OpenAIRE

    Yanping Zhu; Jieru Pan; Junzhi Qiu; Xiong Guan

    2008-01-01

    Entomopathogenic fungus Verticillium lecanii is a promising whitefly and aphid control agent. Chitinases secreted by this insect pathogen have considerable importance in the biological control of some insect pests. An endochitinase gene Vlchit1 from the fungus was cloned and overexpressed in Escherichia coli. The Vlchit1 gene not only contains an open reading frame (ORF) which encodes a protein of 423 amino acids (aa), but also is interrupted by three short introns. A homology modelling of Vl...

  20. Antibacterial activities of Ligaria cuneifolia and Jodina rhombifolia leaf extracts against phytopathogenic and clinical bacteria.

    Science.gov (United States)

    Soberón, José R; Sgariglia, Melina A; Dip Maderuelo, María R; Andina, María L; Sampietro, Diego A; Vattuone, Marta A

    2014-11-01

    Six plant extracts prepared from Ligaria cuneifolia and Jodina rhombifolia were screened for their potential antimicrobial activities against phytopathogens and clinically standard reference bacterial strains. Bioautography and broth microdilution methods were used to study samples antibacterial activities against 7 bacterial strains. The minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of samples were attained. An antibacterial activity guided isolation and identification of active compounds was carried out for L. cuneifolia methanolic extract (LCME). Both methanolic and aqueous extracts from L. cuneifolia showed inhibitory activities against phytopathogenic bacteria, with MICs ranging from 2.5 to 156 μg mL(-1) for LCME and 5 mg mL(-1) for the aqueous extract. None of the three J. rhombifolia extracts showed significant antibacterial activities against phytopathogenic strains (MIC > 5 mg mL(-1)), except for the aqueous extracts against Pseudomonas syringae (MIC = 312 μg mL(-1)). Only LCME showed bactericidal activities against phytopathogenic strains (MBCs = 78 μg mL(-1)). The LCME exhibited significant inhibitory activity against reference clinical strains: Escherichia coli (MIC = 156 μg mL(-1)) and Staphylococcus aureus (MIC = 78 μg mL(-1), MBC = 312 μg mL(-1)). LCME active compounds were identified as flavonol mono and diglycosides, and gallic acid. The antibacterial activity of purified compounds was also evaluated. A synergistic effect against S. aureus was found between gallic acid and a quercetin glycoside. Hence, anti-phytopathogenic bacteria potential compounds isolated from L. cuneifolia could be used as an effective source against bacterial diseases in plants. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  1. In Vitro and In Vivo Tests for Parasitism of Verticillium psalliotae Treschow on Hemileia Vastatrix BERK. and BR.

    Directory of Open Access Journals (Sweden)

    Mahfud, M. C.

    2006-01-01

    Full Text Available The fungus Verticillium psalliotae Treschow is known to grow on the rust sori masses of Hemileia vastatrix in coffee plantings in Malaysia. However, published reports on this parasite or other biocontrol organisms present naturally in the coffee ecosystems in Malaysia are lacking. In vitro and in vivo studies were conducted to elucidate evidence of parasitism of V. psalliotae on H. vastatrix. The host-mycoparasite interaction conducted in vitro resulted a significant reduction in germination of Hemileia uredespores and an increase in germination of Verticillium conidia. A significant reduction in leaf rust severity by the presence of V. psalliotae, particularly when V. psalliotae was applied 24 hours before the inoculation of H. vastatrix, demonstrated an evidence parasitism in vivo. The evidences were supported by SEM observation of penetration and damage to Hemileia uredospores by hyphae of V. psalliotae.

  2. Identification of miRNAs and Their Targets in Cotton Inoculated with Verticillium dahliae by High-Throughput Sequencing and Degradome Analysis

    Directory of Open Access Journals (Sweden)

    Yujuan Zhang

    2015-06-01

    Full Text Available MicroRNAs (miRNAs are a group of endogenous small non-coding RNAs that play important roles in plant growth, development, and stress response processes. Verticillium wilt is a vascular disease in plants mainly caused by Verticillium dahliae Kleb., the soil-borne fungal pathogen. However, the role of miRNAs in the regulation of Verticillium defense responses is mostly unknown. This study aimed to identify new miRNAs and their potential targets that are involved in the regulation of Verticillium defense responses. Four small RNA libraries and two degradome libraries from mock-infected and infected roots of cotton (both Gossypium hirsutum L. and Gossypium barbadense L. were constructed for deep sequencing. A total of 140 known miRNAs and 58 novel miRNAs were identified. Among the identified miRNAs, many were differentially expressed between libraries. Degradome analysis showed that a total of 83 and 24 genes were the targets of 31 known and 14 novel miRNA families, respectively. Gene Ontology analysis indicated that many of the identified miRNA targets may function in controlling root development and the regulation of Verticillium defense responses in cotton. Our findings provide an overview of potential miRNAs involved in the regulation of Verticillium defense responses in cotton and the interactions between miRNAs and their corresponding targets. The profiling of these miRNAs lays the foundation for further understanding of the function of small RNAs in regulating plant response to fungal infection and Verticillium wilt in particular.

  3. Expression of baculovirus anti-apoptotic genes p35 and op-iap in cotton (Gossypium hirsutum L. enhances tolerance to verticillium wilt.

    Directory of Open Access Journals (Sweden)

    Juan Tian

    Full Text Available BACKGROUND: Programmed cell death plays an important role in mediating plant adaptive responses to the environment such as the invasion of pathogens. Verticillium wilt, caused by the necrotrophic pathogen Verticillium dahliae, is a serious vascular disease responsible for great economic losses to cotton, but the molecular mechanisms of verticillium disease and effective, safe methods of resistance to verticillium wilt remain unexplored. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we introduced baculovirus apoptosis inhibitor genes p35 and op-iap into the genome of cotton via Agrobacterium-mediated transformation and analyzed the response of transgenic plants to verticillium wilt. Results showed that p35 and op-iap constructs were stably integrated into the cotton genome, expressed in the transgenic lines, and inherited through the T(3 generation. The transgenic lines had significantly increased tolerance to verticillium wilt throughout the developmental stages. The disease index of T(1-T(3 generation was lower than 19, significantly (P<0.05 better than the negative control line z99668. After treatment with 250 mg/L VD-toxins for 36 hours, DNA from negative control leaves was fragmented, whereas fragmentation in the transgenic leaf DNA did not occur. The percentage of cell death in transgenic lines increased by 7.11% after 60 mg/L VD-toxin treatment, which was less than that of the negative control lines's 21.27%. This indicates that p35 and op-iap gene expression partially protects cells from VD-toxin induced programmed cell death (PCD. CONCLUSION/SIGNIFICANCE: Verticillium dahliae can trigger plant cells to die through induction of a PCD mechanism involved in pathogenesis. This paper provides a potential strategy for engineering broad-spectrum necrotrophic disease resistance in plants.

  4. Genetic structure, linkage disequilibrium and association mapping of Verticillium wilt resistance in elite cotton (Gossypium hirsutum L.) germplasm population.

    Science.gov (United States)

    Zhao, Yunlei; Wang, Hongmei; Chen, Wei; Li, Yunhai

    2014-01-01

    Understanding the population structure and linkage disequilibrium in an association panel can effectively avoid spurious associations and improve the accuracy in association mapping. In this study, one hundred and fifty eight elite cotton (Gossypium hirsutum L.) germplasm from all over the world, which were genotyped with 212 whole genome-wide marker loci and phenotyped with an disease nursery and greenhouse screening method, were assayed for population structure, linkage disequilibrium, and association mapping of Verticillium wilt resistance. A total of 480 alleles ranging from 2 to 4 per locus were identified from all collections. Model-based analysis identified two groups (G1 and G2) and seven subgroups (G1a-c, G2a-d), and differentiation analysis showed that subgroup having a single origin or pedigree was apt to differentiate with those having a mixed origin. Only 8.12% linked marker pairs showed significant LD (Pmapping, which widely were distributed among 15 chromosomes. Among which 10 marker loci were found to be consistent with previously identified QTLs and 32 were new unreported marker loci, and QTL clusters for Verticillium wilt resistanc on Chr.16 were also proved in our study, which was consistent with the strong linkage in this chromosome. Our results would contribute to association mapping and supply the marker candidates for marker-assisted selection of Verticillium wilt resistance in cotton.

  5. Plant carbohydrate scavenging through tonB-dependent receptors: a feature shared by phytopathogenic and aquatic bacteria

    National Research Council Canada - National Science Library

    Blanvillain, Servane; Meyer, Damien; Boulanger, Alice; Lautier, Martine; Guynet, Catherine; Denancé, Nicolas; Vasse, Jacques; Lauber, Emmanuelle; Arlat, Matthieu

    2007-01-01

    .... Analysis of the genome of the phytopathogenic bacterium Xanthomonas campestris pv. campestris (Xcc), predicts 72 TBDRs. Such an overrepresentation is common in Xanthomonas species but is limited to only a small number of bacteria...

  6. Genome Sequence of the Endophytic Bacterium Bacillus thuringiensis Strain KB1, a Potential Biocontrol Agent against Phytopathogens

    OpenAIRE

    Jeong, Haeyoung; Jo, Sung Hee; Hong, Chi Eun; Park, Jeong Mee

    2016-01-01

    Bacillus thuringiensis is the most widely known microbial pesticide used in agricultural applications. Herein, we report a draft genome sequence of the endophytic bacterium Bacillus thuringiensis strain KB1, which exhibits antagonism against phytopathogens.

  7. Genome Sequence of the Endophytic Bacterium Bacillus thuringiensis Strain KB1, a Potential Biocontrol Agent against Phytopathogens.

    Science.gov (United States)

    Jeong, Haeyoung; Jo, Sung Hee; Hong, Chi Eun; Park, Jeong Mee

    2016-04-21

    ITALIC! Bacillus thuringiensisis the most widely known microbial pesticide used in agricultural applications. Herein, we report a draft genome sequence of the endophytic bacterium ITALIC! Bacillus thuringiensisstrain KB1, which exhibits antagonism against phytopathogens.

  8. Island cotton Gbve1 gene encoding a receptor-like protein confers resistance to both defoliating and non-defoliating isolates of Verticillium dahliae.

    Directory of Open Access Journals (Sweden)

    Baolong Zhang

    Full Text Available Verticillium wilt caused by soilborne fungus Verticillium dahliae could significantly reduce cotton yield. Here, we cloned a tomato Ve homologous gene, Gbve1, from an island cotton cultivar that is resistant to Verticillium wilt. We found that the Gbve1 gene was induced by V. dahliae and by phytohormones salicylic acid, jasmonic acid, and ethylene, but not by abscisic acid. The induction of Gbve1 in resistant cotton was quicker and stronger than in Verticillium-susceptible upland cotton following V. dahliae inoculation. Gbve1 promoter-driving GUS activity was found exclusively in the vascular bundles of roots and stems of transgenic Arabidopsis. Virus-induced silencing of endogenous genes in resistant cotton via targeting a fragment of the Gbve1 gene compromised cotton resistance to V. dahliae. Furthermore, we transformed the Gbve1 gene into Arabidopsis and upland cotton through Agrobacterium-mediated transformation. Overexpression of the Gbve1 gene endowed transgenic Arabidopsis and upland cotton with resistance to high aggressive defoliating and non-defoliating isolates of V. dahliae. And HR-mimic cell death was observed in the transgenic Arabidopsis. Our results demonstrate that the Gbve1 gene is responsible for resistance to V. dahliae in island cotton and can be used for breeding cotton varieties that are resistant to Verticillium wilt.

  9. Chemical Composition and Antibacterial Activity of Essential Oils of Two Species of Lamiaceae against Phytopathogenic Bacteria.

    Science.gov (United States)

    Gormez, Arzu; Bozari, Sedat; Yanmis, Derya; Gulluce, Medine; Sahin, Fikrettin; Agar, Guleray

    2015-01-01

    In this study, we aimed to determine chemical composition and antibacterial activities of Satureja hortensis and Calamintha nepeta against to 20 phytopathogenic bacteria causing serious crop loss. The essential oils of S. hortensis and C. nepeta were isolated by the hydrodistillation method and the chemical composition of the essential oils were analyzed by GC-MS. The antibacterial properties of the essential oils were evaluated against 20 phytopathogenic bacteria through Disc diffusion assay and micro dilution assay. The results revealed that the essential oils of S. hortensis and C. nepeta have significant antibacterial activity. Furthermore, the findings of the study are valuable for future investigations focusing on the alternative natural compounds to control plant diseases.

  10. [Cellulase and xylanase activity of phytopathogenic and endophytic fungal strains of Alternaria alternata (Fr.) Keissler].

    Science.gov (United States)

    Kurchenko, I M; Sokolova, O V; Zhdanova, N M; Iarynchyn, A M; Iovenko, O M

    2008-01-01

    A comparative analysis of cellulase and xylanase activity of 25 fungal strains of phytopathogenic and endophytic Alternaria alternata had been realized for the first time using the qualitative reactions. The rate of their linear growth on the media with carboxymethylcellulose or xylane had been studied. The cellulase and xylanase activities clearly depended on the distinct strain. The absence of distinct dependence of cellulase and xylanase activities on the species and organs of host plants was demonstrated. The majority of investigated strains of A. alternata did not possess a cellulase activity or the latter was low, but as a whole the phytopathogenic strains were more active than endophytic ones. Xylanase activity was considerable for the fungal strains of all trophyc groups. It was shown that the level of xylanase activity cannot become a biochemical marker of the A. alternata isolate pathogenicity.

  11. Differences in stability of seed-associated microbial assemblages in response to invasion by phytopathogenic microorganisms

    OpenAIRE

    Samir Rezki; Claire Campion; Beatrice Iacomi-Vasilescu; Anne Preveaux; Youness Toualbia; Sophie Bonneau; Martial Briand; Emmanuelle Laurent; Gilles Hunault; Philippe Simoneau; Marie-Agnès Jacques; Matthieu Barret

    2016-01-01

    Seeds are involved in the vertical transmission of microorganisms from one plant generation to another and consequently act as reservoirs for the plant microbiota. However, little is known about the structure of seed-associated microbial assemblages and the regulators of assemblage structure. In this work, we have assessed the response of seed associated microbial assemblages of Raphanus sativus to invading phytopathogenic agents, the bacterial strain Xanthomonas campestris pv. campestris (Xc...

  12. Antifungal activity of essential oil isolated from Ocimum gratissimum L. (eugenol chemotype) against phytopathogenic fungi

    OpenAIRE

    Terezinha de Jesus Faria; Rafael Sottero Ferreira; Lidiane Yassumoto; José Roberto Pinto de Souza; Noemia Kazue Ishikawa; Aneli de Melo Barbosa

    2006-01-01

    An investigation of antifungal activity of the essential oil obtained by steam-distillation (1.1% w/w) of the aerial parts of Ocimum gratissimum and of an ethanolic extract from the steam-distillation residue was carried out using the agar diffusion method. The results revealed that the essential oil inhibited the growth of all fungi tested, including the phytopathogens, Botryosphaeria rhodina, Rhizoctonia sp. and two strains of Alternaria sp., while the extract from the residue was inactive....

  13. Protoplast transformation as a potential platform for exploring gene function in Verticillium dahliae.

    Science.gov (United States)

    Rehman, Latifur; Su, Xiaofeng; Guo, Huiming; Qi, Xiliang; Cheng, Hongmei

    2016-07-26

    Large efforts have focused on screening for genes involved in the virulence and pathogenicity of Verticillium dahliae, a destructive fungal pathogen of numerous plant species that is difficult to control once the plant is infected. Although Agrobacterium tumefaciens-mediated transformation (ATMT) has been widely used for gene screening, a quick and easy method has been needed to facilitate transformation. High-quality protoplasts, with excellent regeneration efficiency (65 %) in TB3 broth (yeast extract 30 g, casamino acids 30 g and 200g sucrose in 1L H20), were generated using driselase (Sigma D-9515) and transformed with the GFP plasmid or linear GFP cassette using PEG or electroporation. PEG-mediated transformation yielded 600 transformants per microgram DNA for the linear GFP cassette and 250 for the GFP plasmid; electroporation resulted in 29 transformants per microgram DNA for the linear GFP cassette and 24 for the GFP plasmid. To determine whether short interfering RNAs (siRNAs) can be delivered to the protoplasts and used for silencing genes, we targeted the GFP gene of Vd-GFP (V. dahliae GFP strain obtained in this study) by delivering one of four different siRNAs-19-nt duplex with 2-nt 3' overhangs (siRNA-gfp1, siRNA-gfp2, siRNA-gfp3 and siRNA-gfp4)-into the Vd-GFP protoplasts using PEG-mediated transformation. Up to 100 % silencing of GFP was obtained with siRNA-gfp4; the other siRNAs were less effective (up to 10 % silencing). Verticillium transcription activator of adhesion (Vta2) gene of V. dahliae was also silenced with four siRNAs (siRNA-vta1, siRNA-vta2, siRNA-vta3 and siRNA-vta4) independently and together using the same approach; siRNA-vta1 had the highest silencing efficiency as assessed by colony diameter and quantitative real time PCR (qRT-PCR) analysis. Our quick, easy transformation method can be used to investigate the function of genes involved in growth, virulence and pathogenicity of V. dahliae.

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

    Science.gov (United States)

    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.

  15. QTL Analysis of Fiber Yield and Quality and Resistance to Verticillium Wilt Using Gossypium hirsutum and G.barbadense Advanced Backcross Populations

    Institute of Scientific and Technical Information of China (English)

    LI Ai-guo; WANG Tao; YUAN You-lu; LIU Guang-ping; ZHANG Bao-cai; LI Jun-wen; SHI Yu-zhen; LIU Ai-ying; YANG Ze-mao; LIU Zhi; YU Xiao-nan

    2008-01-01

    @@ To introgress elite QTL alleles of Gossypium barbadense L.for fiber yield and quality and resistance to Verticillium wilt into G.hirsutum L.,enlarge the genetic base of G.hirsutum,and provide new germplasm resources for the variety development,the cultivars Zhongmiansuo 36,G.hirsutum,and Hai 1,G.barbadense,were used as recurrent and donor parent,respectively,to produce BC1 F1,BC1S1,and BC2F1 populations.QTL analyses of fiber quality,yield-related,Key words: interspecific backcrosss; AB-QTL; fiber quality; yield; Verticillium wilt resistance

  16. Development of modified medium for the enhancement in antifungal activity of P. steckii (MF1 mangrove fungi against verticillium wilt pathogenic fungi of rose

    Directory of Open Access Journals (Sweden)

    Jyotasanmayee Sabat

    2009-08-01

    Full Text Available The antifungal activity of Penicillium steckii (MF1 against pathogenic fungi of rose causing verticillium wilt was evaluated under differential nutrient condition. A suitable and modified medium was obtained in which test fungus exhibited enhanced antifungal activity and produced larger inhibition zone against the verticillium fungus. The study carried out to modify the medium components at level of nitrogen, carbon source and their concentration on agar plates. Addition of amino acids, metals and plant growth hormones did not show positive effect but glycerol enhanced the antifungal activity very much.

  17. [Construction and analysis of SSH library of Gossypium barbadense upon infection with Verticillium dahliae].

    Science.gov (United States)

    Zhu, Long-Fu; Tu, Li-Li; Zhang, Xian-Long; Nie, Yi-Chun; Guo, Xiao-Ping; Xia, Qi-Zhong

    2005-05-01

    Roots were collected from the seedlings inoculated with pathogen Verticillium dahliae after 2, 4, 8, 12, 24, 48, 72 and 96 hours for total RNA extraction. The cDNAs from the inoculated seedlings were used as the tester and those from the control seedlings as the driver. SSH method was employed to find the differently expressed cDNAs responding to the pathogen. T/A clone library was constructed containing 534 clones. The cDNA inserts were amplified from the bacterial clones directly with M13 primers by PCR. The size of the products ranged 0.2 - 1.2 kb with an average size of 0.5 kb. The SSH products were dotted on nylon filters, and the positive clones were screened by virtual Northern blotting with probes of the two kinds of initiative cDNAs. Totally 78 clones which were up-regulated and putatively involved in the defense response of G. barbadense were identified and sequenced. Sequence similarity searches were performed with the Blastn and Blastx. Most of them showed high or partial homology to genes or ESTs induced by different stresses in Arabidopsis thaliana and other species,such as the pathogenesis-related 10 family of G. hirsumtum and disease resistance-responsive family protein in Arabidopsis thaliana. The results would be helpful to understand the molecular mechanisms of disease response in cotton.

  18. Biochemical indices of hop resistance to Verticillium albo-atrum and Fusarium sambucinum

    Directory of Open Access Journals (Sweden)

    Jan Piotrowski

    2013-12-01

    Full Text Available The contents of total phenolic compounds, chlorogenic acid and peroxidase activity as well as monophenols to polyphenols ratio were studies in the suckers of the hops as indices of resistance to Verticillium albo-atrum and Fusarium sambucinum. The suckers of hop taken in the early spring from the healthy and infected plots were used in the experiments. As a research material were included cv. 'Northern Brewer' - a wilt tolerant variety, two wild susceptible varieties - cv. 'Lubelski' and cv. 'Brewers Gold', four breeding clones and one male plant. It was found that, 'Northern Brewer' contains more total phenolic compounds, rnonophenols and chlorogenic acid, and in particular considerably higher peroxidase activity as compared to cv. 'Lubelski'. Taking into consideration the contents of these compounds, in the majority of cases, the new breeding clones were similar to the mother variety 'Northern Brewer'. It seems resonable to assume, that the new clones should be more wilt tolerant than varieties and populations cultivated in our country.

  19. Investigating Agrobacterium-mediated transformation of Verticillium albo-atrum on plant surfaces.

    Directory of Open Access Journals (Sweden)

    Claire J Knight

    Full Text Available BACKGROUND: Agrobacterium tumefaciens has long been known to transform plant tissue in nature as part of its infection process. This natural mechanism has been utilised over the last few decades in laboratories world wide to genetically manipulate many species of plants. More recently this technology has been successfully applied to non-plant organisms in the laboratory, including fungi, where the plant wound hormone acetosyringone, an inducer of transformation, is supplied exogenously. In the natural environment it is possible that Agrobacterium and fungi may encounter each other at plant wound sites, where acetosyringone would be present, raising the possibility of natural gene transfer from bacterium to fungus. METHODOLOGY/PRINCIPAL FINDINGS: We investigate this hypothesis through the development of experiments designed to replicate such a situation at a plant wound site. A. tumefaciens harbouring the plasmid pCAMDsRed was co-cultivated with the common plant pathogenic fungus Verticillium albo-atrum on a range of wounded plant tissues. Fungal transformants were obtained from co-cultivation on a range of plant tissue types, demonstrating that plant tissue provides sufficient vir gene inducers to allow A. tumefaciens to transform fungi in planta. CONCLUSIONS/SIGNIFICANCE: This work raises interesting questions about whether A. tumefaciens may be able to transform organisms other than plants in nature, or indeed should be considered during GM risk assessments, with further investigations required to determine whether this phenomenon has already occurred in nature.

  20. Molecular mapping of Verticillium wilt resistance QTL clustered on chromosomes D7 and D9 in upland cotton

    Institute of Scientific and Technical Information of China (English)

    JIANG Feng; ZHAO Jun; ZHOU Lei; GUO WangZhen; ZHANG TianZhen

    2009-01-01

    Verticillium wilt is s destructive disease with international consequences for cotton production. Breeding broad-spectrum resistant cultivars is considered to be one of the most effective means for reducing crop losses. A resistant cotton cultivar, 60182, was crossed with a susceptible cultivar, Junmian 1, to identify markers for Verticillium resistance genes and validate the mode of its inheritance. Genetic segregation analysis for Verticillium wilt resistance was evaluated based upon infected leaf percentage in the seedling stage using major gene-polygene mixed inheritance models and joint analysis of P_1, P_2, F_1, B_1, B_2 and F_2 populations obtained from the cultivar cross. We found that resistance of upland cotton cultivar 60182 to isolates BP2, VD8 and T9, and their isoconcentration mixture was controlled by two major genes with additive-dominance-epistatic effects, and the inheritance of the major gene was dominant. Furthermore, a genetic linkage map was constructed using F_2 segregating population and resistance phenotypic data were obtained using F_(2:3) families inoculated with different isolates and detected in different developmental stages. The genetic linkage map with 139 loci was comprised of 31 linkage groups covering 1165 cM, with an average distance of 8.38 cM between two markers, or 25.89% of the cotton genome length. From 60182, we found 4 QTL on chromosome D7 and 4 QTL on D9 for BP2, 5 QTL on D7 and 9 QTL on D9 for VD8, 4 QTL on D7 and 5 QTL on D9 for T9 and 3 QTL on D7 and 7 QTL on D7 for mixed pathogens. The QTL mapping results revealed that QTL clusters with high contribution rates were screened simultaneously on chromosomes D9 and D7 by multiple interval mapping (CIM), whether from resistance phenotypic data from different developmental stages or for different isolates. The result is consistent with the genetic model of two major genes in 60182 and suggests broad-spectrum resistance to both defoliating isolates of V. dahliae and

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

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Glaring, Mikkel Andreas; Olsson, Stefan

    2017-01-01

    BACKGROUND: 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...

  2. The impact of genotyping-by-sequencing pipelines on SNP discovery and identification of markers associated verticillium wilt resistance in autotetraploid alfalfa (sedicago sativa l.)

    Science.gov (United States)

    Verticillium wilt (VW) of alfalfa is a soilborne disease that causes severe yield loss in alfalfa. To identify molecular markers associated with VW resistance, an integrated framework of genome-wide association study (GWAS) with high-throughput genotyping by sequencing (GBS) was used for mapping lo...

  3. Late summer disease symptoms in western Washington red raspberry fields associated with co-occurrence of Phytophthora rubi, Verticillium dahliae, and Pratylenchus penetrans

    Science.gov (United States)

    60% of the $109 million processed by the U.S. red raspberry industry is in northern Washington. In 2012, late summer disease symptoms were observed in many raspberry fields. These symptoms were initially attributed to Verticillium dahliae, but other soilborne pathogens (Phytophthora rubi, Pratylench...

  4. Genotyping-by-sequencing based genome-wide association studies on Verticillium wilt resistance in heterozygous autotetraploid alfalfa (Medicago sativa L.)

    Science.gov (United States)

    Verticillium wilt (VW) is a fungal disease which causes severe yield loss in alfalfa. The most effective method to control the disease is through the development and use of resistant varieties. Identification of gene loci linked to VW resistance will facilitate breeding for the disease-resistant al...

  5. Evolution of the metabolic and regulatory networks associated with oxygen availability in two phytopathogenic enterobacteria

    Directory of Open Access Journals (Sweden)

    Babujee Lavanya

    2012-03-01

    Full Text Available Abstract Background Dickeya dadantii and Pectobacterium atrosepticum are phytopathogenic enterobacteria capable of facultative anaerobic growth in a wide range of O2 concentrations found in plant and natural environments. The transcriptional response to O2 remains under-explored for these and other phytopathogenic enterobacteria although it has been well characterized for animal-associated genera including Escherichia coli and Salmonella enterica. Knowledge of the extent of conservation of the transcriptional response across orthologous genes in more distantly related species is useful to identify rates and patterns of regulon evolution. Evolutionary events such as loss and acquisition of genes by lateral transfer events along each evolutionary branch results in lineage-specific genes, some of which may have been subsequently incorporated into the O2-responsive stimulon. Here we present a comparison of transcriptional profiles measured using densely tiled oligonucleotide arrays for two phytopathogens, Dickeya dadantii 3937 and Pectobacterium atrosepticum SCRI1043, grown to mid-log phase in MOPS minimal medium (0.1% glucose with and without O2. Results More than 7% of the genes of each phytopathogen are differentially expressed with greater than 3-fold changes under anaerobic conditions. In addition to anaerobic metabolism genes, the O2 responsive stimulon includes a variety of virulence and pathogenicity-genes. Few of these genes overlap with orthologous genes in the anaerobic stimulon of E. coli. We define these as the conserved core, in which the transcriptional pattern as well as genetic architecture are well preserved. This conserved core includes previously described anaerobic metabolic pathways such as fermentation. Other components of the anaerobic stimulon show variation in genetic content, genome architecture and regulation. Notably formate metabolism, nitrate/nitrite metabolism, and fermentative butanediol production, differ between E

  6. A Novel and Effective Streptomyces sp. N2 Against Various Phytopathogenic Fungi.

    Science.gov (United States)

    Xu, Bo; Chen, Wei; Wu, Zhi-ming; Long, Yue; Li, Kun-tai

    2015-11-01

    Phytopathogenic fungi would induce a variety of plant diseases, resulting in a severe reduction of agricultural output. However, the current plant disease control is mainly dependent on the environmentally and healthily hazardous chemical fungicides. Thus, the present work aimed to isolate an effective antagonistic microorganism against various soilborne phytopathogenic fungi. By dual culture with Rhizoctonia solani, a novel Streptomyces specie, Streptomyces sp. N2, was screened out from a total of 167 isolated actinomycetes, which displayed a strong inhibitory effect on R. solani (26.85 ± 1.35 mm of inhibition zone diameter). By means of macroporous resin and silica gel column chromatography coupled with preparative HPLC, an antifungal metabolite (3-methyl-3,5-amino-4-vinyl-2-pyrone, C6H7O2N) was isolated and purified from Streptomyces sp. N2. The bioassay results showed that the purified antifungal metabolite could not only possess a broad-spectrum inhibitory effect on a range of plant pathogenic fungi in vitro (e.g., R. solani, Pyricularia grisea, Fusarium oxysporum f. sp. niveum, F. oxysporum f. sp. vasinfectum, Penicillium italicum, and Colletotrichum gloeosporioides), but also had a significantly effective in vivo biocontrol efficacy on grape fruits anthracnose caused by C. gloeosporioides. Microscopic observation indicated that the antifungal metabolite from Streptomyces sp. N2 would exert its antimicrobial activity by disorganizing the cytoplasmic organelles of phytopathogenic fungi. The above results suggested that Streptomyces sp. N2 was one of promising fungicide for biocontrol of fungal plant diseases, especially due to its broad-spectrum and effective antagonist on various plant pathogens.

  7. Role of brassinosteroids in alleviating toxin-induced stress of Verticillium dahliae on cotton callus growth.

    Science.gov (United States)

    Bibi, Noreen; Ahmed, Imrul Mosaddek; Fan, Kai; Dawood, Muhammad; Li, Feng; Yuan, Shuna; Wang, Xuede

    2017-03-29

    Brassinosteroids are well known to mitigate biotic stresses; however, their role to induce tolerance against Verticillium dahliae is unknown. The current study employed V. dahliae (Vd) toxin as pathogen-free model system to induce stress on cotton callus growth, and its amelioration was investigated using 24-epibrassinolide (EBR). Results revealed that EBR has ameliorative effects against Vd toxin with greater seen effect when callus was treated with EBR prior to its exposure to Vd toxin (pre-EBR treatment) than EBR applied along with Vd toxin simultaneously (co-EBR treatment). Pre-EBR-treated calli remained green, while 65 and 90% callus browning was observed in co-EBR- and Vd toxin-alone-treated callus, respectively. Likewise, the fresh weight of the pre-EBR-treated callus was 52% higher than Vd toxin-alone treatment, whereas this increase was only 23% in co-EBR-treated callus. Meanwhile, EBR treatment of the cotton callus has also increased the contents of chlorophylls a and b, carotenoids, total phenols, flavonoids, soluble sugars, and proteins and increased the activity of enzymes involved in secondary metabolism like polyphenol oxidase (PPO), phenylalanine ammonialyase (PAL), cinnamyl alchol dehydrogenase (CAD), and shikimate dehydrogenase (SKDH) over Vd toxin-alone treatment with higher increments being observed in pre-EBR-treated callus. Furthermore, EBR treatment mimicked the DNA damage and improved the structure of mitochondria, granum, stroma thylakoids, and the attachment of ribosomes with the endoplasmic reticulum. This EBR-mediated mitigation was primarily associated with substantially increased contents of photosynthetic pigments and regulation of secondary metabolism.

  8. Verticillium dahliae LysM effectors differentially contribute to virulence on plant hosts.

    Science.gov (United States)

    Kombrink, Anja; Rovenich, Hanna; Shi-Kunne, Xiaoqian; Rojas-Padilla, Eduardo; van den Berg, Grardy C M; Domazakis, Emmanouil; de Jonge, Ronnie; Valkenburg, Dirk-Jan; Sánchez-Vallet, Andrea; Seidl, Michael F; Thomma, Bart P H J

    2017-05-01

    Chitin-binding lysin motif (LysM) effectors contribute to the virulence of various plant-pathogenic fungi that are causal agents of foliar diseases. Here, we report the LysM effectors of the soil-borne fungal vascular wilt pathogen Verticillium dahliae. Comparative genomics revealed three core LysM effectors that are conserved in a collection of V. dahliae strains. Remarkably, and in contrast with the previously studied LysM effectors of other plant pathogens, no expression of core LysM effectors was monitored in planta in a taxonomically diverse panel of host plants. Moreover, targeted deletion of the individual LysM effector genes in V. dahliae strain JR2 did not compromise virulence in infections on Arabidopsis, tomato or Nicotiana benthamiana. Interestingly, an additional lineage-specific LysM effector is encoded in the genome of V. dahliae strain VdLs17, but not in any other V. dahliae strain sequenced to date. Remarkably, this lineage-specific effector is expressed in planta and contributes to the virulence of V. dahliae strain VdLs17 on tomato, but not on Arabidopsis or N. benthamiana. Functional analysis revealed that this LysM effector binds chitin, is able to suppress chitin-induced immune responses and protects fungal hyphae against hydrolysis by plant hydrolytic enzymes. Thus, in contrast with the core LysM effectors of V. dahliae, this lineage-specific LysM effector of strain VdLs17 contributes to virulence in planta. © 2016 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.

  9. Genome-wide identification, phylogeny and expression profile of vesicle fusion components in Verticillium dahliae.

    Directory of Open Access Journals (Sweden)

    Xue Yang

    Full Text Available Vesicular trafficking plays a crucial role in protein localization and movement, signal transduction, and multiple developmental processes in eukaryotic cells. Vesicle fusion is the final and key step in vesicle-mediated trafficking and mainly relies on SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors, the regulators including SM (Sec1/Munc18 family proteins, Rab GTPases and exocyst subunits. Verticillium dahliae is a widespread soil fungus that causes disruptive vascular diseases on a wide range of plants. To date, no genes involved in vesicular fusion process have been identified and characterized in V. dahliae. The recent publication of the draft genome sequence of V. dahliae allowed us to conduct a genome-wide identification, phylogeny and expression profile of genes encoding vesicular fusion components. Using compared genomics and phylogenetic methods, we identified 44 genes encoding vesicle fusion components in the V. dahliae genome. According to the structural features of their encoded proteins, the 44 V. dahliae genes were classified into 22 SNAREs (6 Qa-, 4 Qb-, 6 Qc-, 1 Qbc- and 5 R-types, 4 SM family proteins, 10 Rab GTPases and 8 exocyst proteins. Based on phylogeny and motif constitution analysis, orthologs of vesicle fusion component in filamentous fungi were generally clustered together into the same subclasses with well-supported bootstrap values. Analysis of the expression profiles of these genes indicated that many of them are significantly differentially expressed during vegetative growth and microsclerotia formation in V. dahliae. The analysis show that many components of vesicle fusion are well conserved in filamentous fungi and indicate that vesicle fusion plays a critical role in microsclerotia formation of smoke tree wilt fungus V. dahliae. The genome-wide identification and expression analysis of components involved in vesicle fusion should facilitate research in this gene family and give

  10. Interaction of the psychrotroph Pseudomonas fluorescens In5 with phytopathogens in cold soils

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Olsson, Stefan; Stougaard, Peter

    is disease suppressive owing to the presence of diverse antimicrobial microorganisms. Using culture-based approaches, the psychrotroph Pseudomonas fluorescens In5 was previously isolated from the soil microbiome. The aim of this study is to unravel key biocontrol traits underpinning the antagonistic activity...... of this cold-active bacterium against phytopathogens. Method: A combination of different technologies including genomics, transcriptomics and metabolomics are being used to explore the interaction of the psychrotroph P. fluorescens In5 in dual-culture with diverse plant pathogens. To date, molecular genetics...

  11. [Effect of biopreparations on dynamics of the number of bacteria and phytopathogenic fungi in potato agroecosystem].

    Science.gov (United States)

    Patyka, N V; Borodaĭ, V V; Zhitkevich, N V; Khomenko, E V; Gnatiuk, T T; Koltunov, V A; Patyka, V F

    2012-01-01

    Application of biological preparations such as Phytotsid and Planryz favoures the growth of the general number of soil's bacteria population compared with control by 13.0-36.1% in the variant of potato variety Scarbnytsya and by 4.5-24.6% of potato variety Oberig. It also decreases 1.2-1.8 times the number of soil phytopathogens--Fusarium and Alternaria. During the application of Rovral Akvaflo the Shenon's ecological index of species biodiversity was lower than during the bioprepation application. One could observe a decrease of species biodiversity and dominance of dark pigmentation in fungi--Alternaria sp., Cladosporium sp., Phoma sp., Doratomyces sp., and pigmented bacteria.

  12. Isolation and characterization of microsatellite markers from the phytopathogenic fungus Alternaria dauci.

    Science.gov (United States)

    Benichou, Soumaya; Dongo, Anita; Henni, Djamel Eddine; Peltier, Didier; Simoneau, Philippe

    2009-01-01

    Eleven polymorphic microsatellite markers were isolated from the necrotrophic phytopathogenic fungus Alternaria dauci based on enriched genomic libraries. In order to assess allelic variability, the microsatellite loci were analysed in a collection of 43 isolates. The number of detected alleles in 11 loci ranged from two to 24 (mean 10.4). Test of cross-species amplification and sequencing of the resulting amplicons showed that some of these microsatellites could be used in different species such as Alternaria solani, Alternaria bataticola and Alternaria zinniae. © 2008 The Authors. Journal compilation © 2008 Blackwell Publishing Ltd.

  13. Systemic responses in a tolerant olive (Olea europaea L. cultivar upon root colonization by the vascular pathogen Verticillium dahliae

    Directory of Open Access Journals (Sweden)

    Carmen Gómez-Lama Cabanás

    2015-09-01

    Full Text Available Verticillium wilt of olive (VWO is caused by the vascular pathogen Verticillium dahliae. One of the best VWO management measures is the use of tolerant cultivars; however, our knowledge on VWO tolerance/resistance genetics is very limited. A transcriptomic analysis was conducted to (i identify systemic defence responses induced/repressed in aerial tissues of the tolerant cultivar Frantoio upon root colonization by V. dahliae, and (ii determine the expression pattern of selected defence genes in olive cultivars showing differential susceptibility to VWO. Two suppression subtractive hybridization cDNA libraries, enriched in up-regulated (FU and down-regulated (FD genes respectively, were generated from ‘Frantoio’ aerial tissues. Results showed that broad systemic transcriptomic changes are taking place during V. dahliae-‘Frantoio’ interaction. A total of 585 FU and 381 FD unigenes were identified, many of them involved in defence response to (abiotic stresses. Selected genes were then used to validate libraries and evaluate their temporal expression pattern in ‘Frantoio’. Four defence genes were analyzed in cultivars Changlot Real (tolerant and Picual (susceptible. An association between GRAS1 and DRR2 gene expression patterns and susceptibility to VWO was observed, suggesting that these transcripts could be further evaluated as markers of the tolerance level of olive cultivars to V. dahliae.

  14. Verticase:a Fibrinolytic Enzyme Produced by Verticillium sp.Tj33,an Endophyte of Trachelospermum jasminoides

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Plant endophytes are among the most important resources of biologically active metabolites.Twenty-three endophyte strains residing in Trachelospermum jasminoides were cultivated In with the cultures assayed for the fibrinolytic substance production.As a result,the culture of Verticillium sp.Tj33 was shown to be the most active.A fibrinolytic enzyme designated as verticase was subsequently purified from the supernatant of Verticillium sp.culture broth by a combination of DEAE-52,Sephadex G-75 and hydrophobic column chromatographies.Verticase,with its molecular mass of 31 kDa and pl of 8.5,was demonstrated to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing electrophoresis.Verticase is an enzyme that hydrolyzes fibrin directly without activation of plaminogen.It was stable in a broad pH range from 4 through to 11 with the optimal reaction pH value and temperature shown to be around 9-10 and 50-60℃,respectively.The fibrinolytic activity of verticase was severely inhibited by phenylmethylsulfony fluoride,Indicating that verticase was a serine protease.

  15. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7

    Directory of Open Access Journals (Sweden)

    M. Mercedes eMaldonado-González

    2015-04-01

    Full Text Available The effective management of Verticillium wilts, 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 Verticillium wilt 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 (ND 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 Verticillium wilt (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. Arabidopsis thaliana is therefore a suitable alternative to olive bioassays to unravel biocontrol traits involved in biological control of V. dahliae by P. fluorescens PICF7.

  16. Antifungal potential of Bacillus vallismortis R2 against different phytopathogenic fungi

    Energy Technology Data Exchange (ETDEWEB)

    Kaur, P.K.; Kaur, J.; Saini, H.S.

    2015-07-01

    The cash crops grown in an agro-climatic region are prone to infection by various fungal pathogens. The use of chemical fungicides over the years has resulted in emergence of resistant fungal strains, thereby necessitating the development of effective and environmental friendly alternatives. The natural antagonistic interactions among different microbial populations have been exploited as an eco-friendly approach for controlling fungal pathogens resistant to synthetic chemicals. Morphologically distinct bacterial cultures (150), isolated from rhizospheric soils of wheat, rice, onion and tomato plants were screened for their antifungal potential against seven phytopathogenic fungi prevalent in the State of Punjab (India). The bacterial isolate R2, identified as Bacillus vallismortis, supported more than 50% inhibition of different phytopathogenic fungi (Alternaria alternata, Rhizoctonia oryzae, Fusarium oxysporum, Fusarium moniliforme, Colletotrichum sp, Helminthosporium sp and Magnaporthe grisea) in dual culture plate assay. The thin layer chromatography based bio-autography of acid-precipitated biomolecules (APB) indicated the presence of more than one type of antifungal molecule, as evidenced from zones of inhibition against the respective fungal pathogen. The initial analytical studies indicated the presence of surfactin, iturin A and fengycin-like compounds in APB. The antifungal activity of whole cells and APB of isolate R2 was evaluated by light and scanning electron microscopy. The wheat grains treated with APB and exposed to spores of A. alternata showed resistance to the development of black point disease, thereby indicating the potential application of R2 and its biomolecules at field scale level. (Author)

  17. In Vitro Antifungal Activity of Sanguinarine and Chelerythrine Derivatives against Phytopathogenic Fungi

    Directory of Open Access Journals (Sweden)

    Yan-Ni Ma

    2012-11-01

    Full Text Available In order to understand the antifungal activity of some derivatives of sanguinarine (S and chelerythrine (C and their structure-activity relationships, sixteen derivatives of S and C were prepared and evaluated for in vitro antifungal activity against seven phytopathogenic fungi by the mycelial growth rate method. The results showed that S, C and their 6-alkoxy dihydro derivatives S1–S4, C1–C4 and 6-cyanodihydro derivatives S5, C5 showed significant antifungal activity at 100 µg/mL against all the tested fungi. For most tested fungi, the median effective concentrations of S, S1, C and C1 were in a range of 14–50 µg/mL. The structure-activity relationship showed that the C=N+ moiety was the determinant for the antifungal activity of S and C. S1–S5 and C1–C5 could be considered as the precursors of S and C, respectively. Thus, the present results strongly suggested that S and C or their derivatives S1–S5 and C1–C5 should be considered as good lead compounds or model molecules to develop new anti-phytopathogenic fungal agents.

  18. The DNA damage response signaling cascade regulates proliferation of the phytopathogenic fungus Ustilago maydis in planta.

    Science.gov (United States)

    de Sena-Tomás, Carmen; Fernández-Álvarez, Alfonso; Holloman, William K; Pérez-Martín, José

    2011-04-01

    In the phytopathogenic fungus Ustilago maydis, the dikaryotic state dominates the period of growth occurring during the infectious phase. Dikaryons are cells in which two nuclei, one from each parent cell, share a single cytoplasm for a period of time without undergoing nuclear fusion. In fungal cells, maintenance of the dikaryotic state requires an intricate cell division process that often involves the formation of a structure known as the clamp connection as well as the sorting of one of the nuclei to this structure to ensure that each daughter dikaryon inherits a balance of each parental genome. Here, we describe an atypical role of the DNA damage checkpoint kinases Chk1 and Atr1 during pathogenic growth of U. maydis. We found that Chk1 and Atr1 collaborate to control cell cycle arrest during the induction of the virulence program in U. maydis and that Chk1 and Atr1 work together to control the dikaryon formation. These findings uncover a link between a widely conserved signaling cascade and the virulence program in a phytopathogen. We propose a model in which adjustment of the cell cycle by the Atr1-Chk1 axis controls fidelity in dikaryon formation. Therefore, Chk1 and Atr1 emerge as critical cell type regulators in addition to their roles in the DNA damage response.

  19. Antifungal potential of Bacillus vallismortis R2 against different phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    Preet K. Kaur

    2015-06-01

    Full Text Available The cash crops grown in an agro-climatic region are prone to infection by various fungal pathogens. The use of chemical fungicides over the years has resulted in emergence of resistant fungal strains, thereby necessitating the development of effective and environmental friendly alternatives. The natural antagonistic interactions among different microbial populations have been exploited as an eco-friendly approach for controlling fungal pathogens resistant to synthetic chemicals. Morphologically distinct bacterial cultures (150, isolated from rhizospheric soils of wheat, rice, onion and tomato plants were screened for their antifungal potential against seven phytopathogenic fungi prevalent in the State of Punjab (India. The bacterial isolate R2, identified as Bacillus vallismortis, supported more than 50% inhibition of different phytopathogenic fungi (Alternaria alternata, Rhizoctonia oryzae, Fusarium oxysporum, Fusarium moniliforme, Colletotrichum sp, Helminthosporium sp and Magnaporthe griseain dual culture plate assay. The thin layer chromatography based bio-autography of acid-precipitated biomolecules (APB indicated the presence of more than one type of antifungal molecule, as evidenced from zones of inhibition against the respective fungal pathogen. The initial analytical studies indicated the presence of surfactin, iturin A and fengycin-like compounds in APB. The antifungal activity of whole cells and APB of isolate R2 was evaluated by light and scanning electron microscopy. The wheat grains treated with APB and exposed to spores of A. alternata showed resistance to the development of black point disease, thereby indicating the potential application of R2 and its biomolecules at field scale level.

  20. Role of iron homeostasis in the virulence of phytopathogenic bacteria: an 'à la carte' menu.

    Science.gov (United States)

    Franza, Thierry; Expert, Dominique

    2013-05-01

    The interaction between pathogenic microbes and their hosts is determined by survival strategies on both sides. As a result of its redox properties, iron is vital for the growth and proliferation of nearly all organisms, including pathogenic bacteria. In bacteria-vertebrate interactions, competition for this essential metal is critical for the outcome of the infection. The role of iron in the virulence of plant pathogenic bacteria has only been explored in a few pathosystems in the past. However, in the last 5 years, intensive research has provided new insights into the mechanisms of iron homeostasis in phytopathogenic bacteria that are involved in virulence. This review, which includes important plant pathosystems, discusses the recent advances in the understanding of iron transport and homeostasis during plant pathogenesis. By summarizing the recent progress, we wish to provide an updated view clarifying the various roles played by this metal in the virulence of bacterial phytopathogens as a nutritional and regulatory element. The complex intertwining of iron metabolism and oxidative stress during infection is emphasized.

  1. Multiple effects of Bacillus amyloliquefaciens volatile compounds: plant growth promotion and growth inhibition of phytopathogens.

    Science.gov (United States)

    Asari, Shashidar; Matzén, Staffan; Petersen, Mikael Agerlin; Bejai, Sarosh; Meijer, Johan

    2016-06-01

    Biotic interactions through volatile organic compounds (VOC) are frequent in nature. This investigation aimed to study the role of ITALIC! BacillusVOC for the beneficial effects on plants observed as improved growth and pathogen control. Four ITALIC! Bacillus amyloliquefacienssubsp. ITALIC! plantarumstrains were screened for VOC effects on ITALIC! Arabidopsis thalianaCol-0 seedlings and ITALIC! Brassicafungal phytopathogens. VOC from all four ITALIC! Bacillusstrains could promote growth of ITALIC! Arabidopsisplants resulting in increased shoot biomass but the effects were dependent on the growth medium. Dose response studies with UCMB5113 on MS agar with or without root exudates showed significant plant growth promotion even at low levels of bacteria. ITALIC! BacillusVOC antagonized growth of several fungal pathogens ITALIC! in vitro However, the plant growth promotion efficacy and fungal inhibition potency varied among the ITALIC! Bacillusstrains. VOC inhibition of several phytopathogens indicated efficient microbial antagonism supporting high rhizosphere competence of the ITALIC! Bacillusstrains. GC-MS analysis identified several VOC structures where the profiles differed depending on the growth medium. The ability of ITALIC! Bacillusstrains to produce both volatile and soluble compounds for plant growth promotion and disease biocontrol provides examples of rhizosphere microbes as an important ecosystem service with high potential to support sustainable crop production.

  2. Effect of anaerobic soil disinfestation and vermicompost on soilborne phytopathogenic agents under tree-crop nursery conditions

    Science.gov (United States)

    Anaerobic soil disinfestation (ASD) is a fumigation-independent management strategy for controlling soilborne pathogens. Walnut nurseries currently employ preplant fumigation to control soilborne phytopathogens and weeds, and may be amenable to use ASD instead. We investigated the potential of ASD a...

  3. Antimicrobial activity of essential oils of Thymus vulgaris and Origanum vulgare on phytopathogenic strains isolated from soybean.

    Science.gov (United States)

    Oliva, M de las M; Carezzano, M E; Giuliano, M; Daghero, J; Zygadlo, J; Bogino, P; Giordano, W; Demo, M

    2015-05-01

    The aim of this work was to study the antimicrobial activity of essential oils obtained from Thymus vulgaris (thyme) and Origanum vulgare (oregano) on phytopathogenic Pseudomonas species isolated from soybean. Strains with characteristics of P. syringae were isolated from leaves of soybean plants with blight symptoms. Ten of these could be identified in Group Ia of LOPAT as P. syringae. Six of these were confirmed as P. syringae using 16S rRNA, indicating the presence of these phytopathogenic bacteria in east and central Argentina. All the phytopathogenic bacteria were re-isolated and identified from the infected plants. MIC values for thyme were 11.5 and 5.7 mg·ml(-1) on P. syringae strains, while oregano showed variability in the inhibitory activity. Both essential oils inhibited all P. syringae strains, with better inhibitory activity than the antibiotic streptomycin. The oils were not bactericidal for all pseudomonads. Both oils contained high carvacrol (29.5% and 19.7%, respectively) and low thymol (1.5%). Natural products obtained from aromatic plants represent potential sources of molecules with biological activity that could be used as new alternatives for the treatment of phytopathogenic bacteria infections.

  4. Soil metagenomics to identify novel mechanisms of antagonism and antifungal activity for the improved control of phytopathogens

    NARCIS (Netherlands)

    Overbeek, van L.S.; Smalla, K.; Bailey, M.; Jansson, J.; Sjöling, S.; Vogel, T.; Nalin, R.; Elsas, van J.D.

    2008-01-01

    Objective: This project will explore & exploit the anti-phytopathogen diversity present in selected European suppressive soils representing a wide geographical spread. The untapped functional potential present in the unculturable part of the microbiota of these soils will be accessed using metag

  5. A comparison of real-time PCR protocols for the quantitative monitoring of asymptomatic olive infections by Verticillium dahliae pathotypes.

    Science.gov (United States)

    Gramaje, D; Pérez-Serrano, V; Montes-Borrego, M; Navas-Cortés, J A; Jiménez-Díaz, R M; Landa, B B

    2013-10-01

    Early, specific, and accurate in planta detection and quantification of Verticillium dahliae are essential to prevent the spread of Verticillium wilt in olive using certified pathogen-free planting material and development of resistance. We comparatively assessed the accuracy, specificity, and efficiency of eight real-time quantitative polymerase chain reaction protocols published since 2002 for the specific detection and quantification of V. dahliae in various host plant species and in soil, using a background of DNAs extracted from olive roots, stems, and leaves. Results showed that some of those protocols were not specific for V. dahliae or were inhibited when using backgrounds other than water. Ranking of protocols according to a weighted score system placed protocols TAQ (based on intergenic spacer ribosomal DNA target gene) and SYBR-4 (based on the β-tubulin 2 target gene) first in sensitivity and efficiency for the quantification of V. dahliae DNA in small amounts and different types of olive tissues (root and stem) tested. Use of TAQ and SYBR-4 protocols allowed accurate quantification of V. dahliae DNA regardless of the background DNA, with a detection limit being fixed at a cycle threshold of 36 (≈18 fg for SYBR-4 and 15 fg for TAQ) of V. dahliae. The amount of DNA from defoliating (D) and nondefoliating (ND) V. dahliae pathotypes was monitored in Verticillium wilt-resistant 'Frantoio' olive using the TAQ and SYBR-4 protocols. In the infection bioassay, higher amounts of D V. dahliae DNA were measured in olive stems, whereas the average amount of fungal DNA in roots was higher for ND-infected plants than D-infected ones. Overall, V. dahliae DNA amounts in all olive tissues tested tended to slightly decrease or remain stable by the end of the experiment (35 days after inoculation). The SYBR-4 and TAQ protocols further enabled detection of V. dahliae in tissues of symptomless plants, suggesting that both techniques can be useful for implementing

  6. Phytopathogenic Bacteria

    NARCIS (Netherlands)

    Wolf, van der J.M.; Boer, de S.H.

    2015-01-01

    A few hundred bacterial species, belonging to the Proteobacteria, Mollecutes and Actinomycetes cause a large number of different plant diseases, some of which are devastating for agricultural crops. Symptoms of bacterial plant diseases are diverse and include necrosis, tissue maceration, wilting, an

  7. Phytopathogenic Bacteria

    NARCIS (Netherlands)

    Wolf, van der J.M.; Boer, de S.H.

    2015-01-01

    A few hundred bacterial species, belonging to the Proteobacteria, Mollecutes and Actinomycetes cause a large number of different plant diseases, some of which are devastating for agricultural crops. Symptoms of bacterial plant diseases are diverse and include necrosis, tissue maceration, wilting,

  8. Comparative pathogenicity, biocontrol efficacy, and multilocus sequence typing of Verticillium nonalfalfae from the invasive Ailanthus altissima and other hosts.

    Science.gov (United States)

    Kasson, M T; Short, D P G; O'Neal, E S; Subbarao, K V; Davis, D D

    2014-03-01

    Verticillium wilt, caused by Verticillium nonalfalfae, is currently killing tens of thousands of highly invasive Ailanthus altissima trees within the forests in Pennsylvania, Ohio, and Virginia and is being considered as a biological control agent of Ailanthus. However, little is known about the pathogenicity and virulence of V. nonalfalfae isolates from other hosts on Ailanthus, or the genetic diversity among V. nonalfalfae from confirmed Ailanthus wilt epicenters and from locations and hosts not associated with Ailanthus wilt. Here, we compared the pathogenicity and virulence of several V. nonalfalfae and V. alfalfae isolates, evaluated the efficacy of the virulent V. nonalfalfae isolate VnAa140 as a biocontrol agent of Ailanthus in Pennsylvania, and performed multilocus sequence typing of V. nonalfalfae and V. alfalfae. Inoculations of seven V. nonalfalfae and V. alfalfae isolates from six plant hosts on healthy Ailanthus seedlings revealed that V. nonalfalfae isolates from hosts other than Ailanthus were not pathogenic on Ailanthus. In the field, 100 canopy Ailanthus trees were inoculated across 12 stands with VnAa140 from 2006 to 2009. By 2011, natural spread of the fungus had resulted in the mortality of >14,000 additional canopy Ailanthus trees, 10,000 to 15,000 Ailanthus sprouts, and nearly complete eradication of Ailanthus from several smaller inoculated stands, with the exception of a few scattered vegetative sprouts that persisted in the understory for several years before succumbing. All V. nonalfalfae isolates associated with the lethal wilt of Ailanthus, along with 18 additional isolates from 10 hosts, shared the same multilocus sequence type (MLST), MLST 1, whereas three V. nonalfalfae isolates from kiwifruit shared a second sequence type, MLST 2. All V. alfalfae isolates included in the study shared the same MLST and included the first example of V. alfalfae infecting a non-lucerne host. Our results indicate that V. nonalfalfae is host adapted and

  9. Anthelmintic efficacy of gold nanoparticles derived from a phytopathogenic fungus, Nigrospora oryzae.

    Directory of Open Access Journals (Sweden)

    Pradip Kumar Kar

    Full Text Available Exploring a green chemistry approach, this study brings to the fore, the anthelmintic efficacy of gold nanoparticles, highlighting the plausible usage of myconanotechnology. Gold nanoparticles of ∼6 to ∼18 nm diameter were synthesized by treating the mycelia-free culture filtrate of the phytopathogenic fungus with gold chloride. Their size and morphology were confirmed by UV-Vis spectroscopy, DLS data, AFM and TEM images. The XRD studies reveal a crystalline nature of the nanoparticles, which are in cubic phase. The FTIR spectroscopic studies before and after the formation of nanoparticles show the presence of possible functional groups responsible for the bio-reduction and capping of the synthesized gold nanoparticles. The latter were tested as vermifugal agents against a model cestode Raillietina sp., an intestinal parasite of domestic fowl. Further, ultrastructural and biochemical parameters were used to corroborate the efficacy study.

  10. "Light-tagged" bacteriophage as a diagnostic tool for the detection of phytopathogens.

    Science.gov (United States)

    Schofield, David; Bull, Carolee T; Rubio, Isael; Wechter, W Patrick; Westwater, Caroline; Molineux, Ian J

    2013-01-01

    Detection of the phytopathogen Pseudomonas cannabina pv alisalensis, the causal agent of bacterial blight of crucifers is essential for managing this disease. A phage-based diagnostic assay was developed that detects and identifies P. cannabina pv alisalensis from cultures and diseased plant specimens. A recombinant "light-tagged" reporter phage was generated by integrating the luxAB genes into the P. cannabina pv alisalensis phage PBSPCA1 genome. PBSPCA1::luxAB is viable, stable and detects P. cannabina pv alisalensis within minutes and with high sensitivity by conferring a bioluminescent signal. Detection is dependent on cell viability since cells treated with a bactericidal disinfectant are unable to elicit a signal. Importantly, the reporter phage detects P. cannabina pv alisalensis from diseased plant specimens indicating the potential of the diagnostic for disease identification. The reporter phage displays promise for the rapid and specific diagnostic detection of cultivated isolates, and infected plant specimens.

  11. A Survey of Phytopathogenic Fungi and Oomycetes in Riyadh, Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Arya Widyawan

    2015-06-01

    Full Text Available A survey of phytopathogenic fungi and Oomyceteswas conducted in Riyadh, Kingdom of Saudi Arabia duringOctober 2008 – May 2009. Total of 223 samples were collectedfrom four regions; Al-Kharj, Oyaynah, Old Diriyah, and Al Amariyah. Isolation was done using Potato Dextrose Agar(PDA. Infected parts were cut then sterilized in chlorox(10%, then were put in petridish that contain PDA andincubated at 25-27 °C. A total twelve genera of fungi andsingle genera of Oomycetes were isolated from the infectedplants and identifi ed as Fusarium spp., Alternaria spp.,Helmintosphorium (Bipolaris spp., Sclerotium spp., Rhizoctoniaspp., Cladosporium spp., Mauginiella scattae, Erysiphe spp.,Leveillula spp., Macrophomina phaseolina, Ustilago spp.,Ulocladium spp., and Phytium spp.

  12. Effect of fragarin on the cytoplasmic membrane of the phytopathogen Clavibacter michiganensis.

    Science.gov (United States)

    Filippone, M P; Diaz-Ricci, J C; Castagnaro, A P; Farías, R N

    2001-07-01

    Fragarin, an antibiotic that was isolated and purified from a soluble fraction of strawberry leaves, may be a new type of preformed antimicrobial compound (phytoanticipin). Here, we report that the growth and oxygen consumption of the phytopathogenic bacterium Clavibacter michiganensis were rapidly inhibited after the addition of fragarin to cultures. Also, dissipation of the membrane potential and an increase of cell membrane permeability were observed in the presence of fragarin. The ability of fragarin to dissipate the membrane potential was confirmed with the use of small unilamellar liposomes made with lipids extracted from C. michiganensis. Our results suggest that fragarin is able to act at the membrane level, and that this action is correlated with a decrease in cell viability.

  13. BIOTRANSFORMATION OF FERULIC ACID BY THE PHYTOPATHOGENIC FUNGI Colletotrichum acutatum AND Lasiodiplodia theobromae

    Directory of Open Access Journals (Sweden)

    Manuel Alejandro Numpaque

    2016-01-01

    Full Text Available The microbial transformation of ferulic acid (FA offers a cleaner, more economical alternative for the natural production of flavorings and fragrances. In the present study, the biotransformation of FA using the filamentous phytopathogenic fungi Colletotrichum acutatum and Lasiodiplodia theobromae was researched. Initially, the toxicity of FA against both fungi was evaluated; the FA displayed a moderate toxicity (total inhibition at concentrations ≥ 2000 mg L-1 and apparently a detoxification mechanism was present. Afterwards, the microorganisms were incubated with the substrate at room conditions using a Czapek-Dox culture medium. The results demonstrated that the FA was mainly converted to 4-vinylguaiacol, reaching the highest abundance within the first 48 hours. To a lesser extent, acetovanillone, ethylguaiacol, and vanillin, among others, were produced. Interestingly, the compounds generated in the biotransformation of FA with C. acutatum and L. theobromae have been used as flavorings. Based on the identified metabolites, a possible metabolic pathway was proposed.

  14. Purification and characterization of beta-N-acetylhexosaminidase from the phytopathogenic fungus Bipolaris sorokiniana.

    Science.gov (United States)

    Geimba, M P; Riffel, A; Brandelli, A

    1998-10-01

    N-acetylhexosaminidase (HEX) from the phytopathogenic fungus Bipolaris sorokiniana was isolated and characterized. The production of HEX by B. sorokiniana was not altered by growing on different carbon sources. Enzyme purification was carried out by sequential liquid chromatography on Sephacryl S-200 HR, and p-aminobenzyl-2-acetamido-2-deoxy-beta-D-thioglucopyranoside agarose. The purification was about 70-fold, with a yield of 41%, determined with p-nitrophenyl-N-acetylglucosaminide as substrate. The enzyme had pH and temperature optima of 4.5 and 55 degrees C, respectively. The molecular weight of non-denatured enzyme was estimated as 120,000 Da by gel filtration chromatography, and about 55,000 Da by SDS-PAGE. The fungal HEX had glycosylated residues as evidenced by binding to Concanavalin-A. Bipolaris sorokiniana enzyme was also active with p-nitrophenyl-chitobioside and p-nitrophenyl-N-acetylgalactosaminide as substrates.

  15. Soil suppressiveness and functional diversity of the soil microflora in organic farming systems

    NARCIS (Netherlands)

    Postma, J.; Schilder, M.T.; Bloem, J.; Leeuwen, van W.K.

    2008-01-01

    Arable fields of 10 organic farms from different locations in The Netherlands were sampled in three subsequent years. The soil samples were analysed for disease suppressiveness against Rhizoctonia solani AG2.2IIIB in sugar beet, Streptomyces scabies in radish and Verticillium longisporum in oilseed

  16. Dysfunctionality of the xylem in Olea europaea L. Plants associated with the infection process by Verticillium dahliae Kleb. Role of phenolic compounds in plant defense mechanism.

    Science.gov (United States)

    Báidez, Ana G; Gómez, Pedro; Del Río, José A; Ortuño, Ana

    2007-05-02

    Xylem ultrastructural modification and the possible participation of phenolic compounds in the natural defense or resistance mechanisms of olive plants infected with Verticillium dahliae Kleb. were studied. Microscopic study showed that the mycelium propagated and passed from one element to another through the pit. The formation of tyloses and aggregates contributed to obstruction of the xylem lumen. In vivo changes in the levels of these phenolic compounds in infected olive plants and their antifungal activity against Verticillium dahliae Kleb., as revealed by in vitro study, strongly suggest that they are involved in natural defense or resistance mechanisms in this plant material, the most active being quercetin and luteolin aglycons, followed by rutin, oleuropein, luteolin-7-glucoside, tyrosol, p-coumaric acid, and catechin. .

  17. Postharvest Survival of Porcine Sapovirus, a Human Norovirus Surrogate, on Phytopathogen-Infected Leafy Greens.

    Science.gov (United States)

    Esseili, Malak A; Chin, Ashlina; Saif, Linda; Miller, Sally A; Qu, Feng; Lewis Ivey, Melanie L; Wang, Qiuhong

    2015-08-01

    Leafy greens are increasingly being recognized as an important vehicle for human noroviruses (HuNoV), which cause recurring gastroenteritis outbreaks. Leafy greens often become infected by phytopathogens in the field, which may cause symptoms on the edible parts. Whether plant pathogen infections enhance the survival of HuNoV on leafy greens is unknown. Lettuce and spinach plants were infected with a bacterium, Xanthomonas campestris pv. vitians strain 701a, and with Cucumber mosaic virus strain Fny, respectively. The survival rate of porcine sapovirus (SaV), a HuNoV surrogate, on infected and noninfected postharvest leaves was then assessed. In addition, acibenzolar-S-methyl, a commercial chemical elicitor of plant systemic defense, was used to assess whether stimulating the plant host defense affects the postharvest survival of SaV. Leaves harvested from control and treated plants were inoculated with SaV and incubated for 7 days at 4°C. The infectivity (tissue culture infectious dose affecting 50% of the culture [TCID50]/ml) and RNA (genomic equivalent/ml) titers of SaV were assayed using immunohistochemistry staining and SaV-specific TaqMan real-time reverse transcription PCR. Our results showed that cucumber mosaic virus Fny induced mild, nonnecrotic symptoms on spinach leaves and had no effect on SaV survival. In contrast, X. campestris pv. vitians 701a induced small localized necrotic lesions and significantly enhanced SaV survival on lettuce leaves. Treatment with acibenzolar-S-methyl was effective in reducing X. campestris pv. vitians 701a-induced lesions on infected lettuce plants but had no direct effect on SaV survival when used on healthy lettuce plants. These findings indicate that phytopathogen-induced necrotic lesions may enhance the postharvest survival of HuNoV on lettuce leaves. Therefore, preventive measures aiming to maintain healthy plants and minimize preharvest biological damage are expected to improve the safety of leafy greens.

  18. In vitro photodynamic inactivation of conidia of the phytopathogenic fungus Colletotrichum graminicola with cationic porphyrins.

    Science.gov (United States)

    Vandresen, Camila Chevonica; Gonçalves, Alan Guilherme; Ducatti, Diogo Ricardo Bazan; Murakami, Fabio Seigi; Noseda, Miguel Daniel; Duarte, Maria Eugenia Rabello; Barreira, Sandra Mara Woranovicz

    2016-05-11

    Photodynamic inactivation (PDI) is an efficient approach for the elimination of a series of microorganisms; however, PDI involving phytopathogenic filamentous fungi is scarce in the literature. In the present study, we have demonstrated the photoinactivating properties of five cationic meso-(1-methyl-4-pyridinio)porphyrins on conidia of the phytopathogen Colletotrichum graminicola. For this purpose, photophysical properties (photostability and (1)O2 singlet production) of the porphyrins under study were first evaluated. PDI assays were then performed with a fluence of 30, 60, 90 and 120 J cm(-2) and varying the porphyrin concentration from 1 to 25 μmol L(-1). Considering the lowest concentration that enabled the best photoinactivation, with the respective lowest effective irradiation time, the meso-(1-methyl-4-pyridinio)porphyrins herein studied could be ranked as follows: triple-charged 4 (1 μmol L(-1) with a fluence of 30 J cm(-2)) > double-charged-trans2 (1 μmol L(-1) with 60 J cm(-2)) > tetra-charged 5 (15 μmol L(-1) with 90 J cm(-2)) > mono-charged 1 (25 μmol L(-1) with 120 J cm(-2)). Double-charged-cis-porphyrin 3 inactivated C. graminicola conidia in the absence of light. Evaluation of the porphyrin binding to the conidia and fluorescence microscopic analysis were also performed, which were in agreement with the PDI results. In conclusion, the cationic porphyrins herein studied were considered efficient photosensitizers to inactivate C. graminicola conidia. The amount and position of positive charges are related to the compounds' amphiphilicity and therefore to their photodynamic activity.

  19. Trichoderma sp Native from Chili Region of Poanas, Durango, Mexico Antagonist against Phytopathogen Fungi

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    Gabriela B. Valencia

    2011-01-01

    Full Text Available Problem statement: Presence of Trichoderma spp. in agricultural soils decrease incidence of diseases by phytopathogen fungi. Sanity diagnostic require to know if exist beneficial microorganism and what agricultural practices help to their propagation. Approach: Samples (30 were taken from soils and sick plants of ten sites in four localities of Valley of Poanas. Phytophthora capsici Leo, Rhizoctonia solani Kuhn and Trichoderma sp were isolated in agar V8 and were identified by microscopy. Results: In the 30 samples analyzed the presence of Phytophthora capsici Leo and Rhizoctonia solani Kuhn was determined. Two isolations of Trichoderma sp were obtained from soil, they had antagonist activity against to P. capsici and R. solani on agar-V8 medium and showed chitinase activity. Sugar production in chitinase (10 mg.mL-1 by crude extract of Trichoderma growth in basal medium more chitin was determined. The average of sugar production from strains were 0.1175 and 0.1125 mg.mL-1 and standard deviations were 0.0567 and 0.0567 in four repetition. Interviews were applied to fifty farmers about cultivars and cultivation practices. At least seven types of chili were cultivated in the region of the Valley of Poanas, inorganic fertilization, irrigation systems by channel, gates and pumps were used. One hundred percent of farmers reported diseases of Damping off and Phytophthora root. Biocides were not used to control these diseases. Conclusion: The natural presence of Trichoderma spp was detected in Valley of Poanas, but some practices as inorganic fertilization and irrigation system can be contributing to propagation of phytopathogen fungi.

  20. Fungal phytopathogens encode functional homologues of plant rapid alkalinization factor (RALF) peptides.

    Science.gov (United States)

    Thynne, Elisha; Saur, Isabel M L; Simbaqueba, Jaime; Ogilvie, Huw A; Gonzalez-Cendales, Yvonne; Mead, Oliver; Taranto, Adam; Catanzariti, Ann-Maree; McDonald, Megan C; Schwessinger, Benjamin; Jones, David A; Rathjen, John P; Solomon, Peter S

    2016-06-13

    In this article, we describe the presence of genes encoding close homologues of an endogenous plant peptide, rapid alkalinization factor (RALF), within the genomes of 26 species of phytopathogenic fungi. Members of the RALF family are key growth factors in plants, and the sequence of the RALF active region is well conserved between plant and fungal proteins. RALF1-like sequences were observed in most cases; however, RALF27-like sequences were present in the Sphaerulina musiva and Septoria populicola genomes. These two species are pathogens of poplar and, interestingly, the closest relative to their respective RALF genes is a poplar RALF27-like sequence. RALF peptides control cellular expansion during plant development, but were originally defined on the basis of their ability to induce rapid alkalinization in tobacco cell cultures. To test whether the fungal RALF peptides were biologically active in plants, we synthesized RALF peptides corresponding to those encoded by two sequenced genomes of the tomato pathogen Fusarium oxysporum f. sp. lycopersici. One of these peptides inhibited the growth of tomato seedlings and elicited responses in tomato and Nicotiana benthamiana typical of endogenous plant RALF peptides (reactive oxygen species burst, induced alkalinization and mitogen-activated protein kinase activation). Gene expression analysis confirmed that a RALF-encoding gene in F. oxysporum f. sp. lycopersici was expressed during infection on tomato. However, a subsequent reverse genetics approach revealed that the RALF peptide was not required by F. oxysporum f. sp. lycopersici for infection on tomato roots. This study has demonstrated the presence of functionally active RALF peptides encoded within phytopathogens that harbour an as yet undetermined role in plant-pathogen interactions.

  1. Synthesis and biological evaluation of arylpyrazoles as fungicides against phytopathogenic fungi.

    Science.gov (United States)

    Zhang, Jin; Peng, Ju-Fang; Wang, Tao; Kang, Yang; Jing, Sisi; Zhang, Zun-Ting

    2017-05-01

    3-phenol-1H-pyrazoles (2), 4-halogeno-3-phenol-1H-pyrazoles (3) and 2-(1-phenol-1H-pyrazol-5-yl)phenols (4) were prepared by the condensation of (E)-3-(dimethylamino)-1-phenylprop-2-en-1-ones and hydrazine hydrate or phenylhydrazine in good yields. They were evaluated against five phytopathogens fungi, namely Cytospora sp., Colletotrichum gloeosporioides, Botrytis cinerea, Alternaria solani and Fusarium solani in vitro. Most of the above-mentioned compounds exhibited activities. For example, 4-chloro-2-(1H-pyrazol-3-yl)phenol (3k) and 4-bromo-3-phenol-1H-pyrazole (3b) showed good and broad-spectrum antifungal properties against Cytospora sp., C. gloeosporioides, Botrytis cinerea, Alternaria solani and F. Solani with [Formula: see text] values ranging from 4.66 to 12.47 [Formula: see text]g/mL. The results showed that pyrazoles with one aryl group at 3-position (2 and 3) exhibited better antibacterial activity than those with two aryl substituents (4). In addition, the existence of an electron-withdrawing group, a substituent on the ortho-position of phenol ring or a halogen atom at the 4-position of the pyrazole enhanced the antifungal activity of pyrazoles 2 and 3. A series of arylpyrazole derivatives was facilely prepared and was evaluated against five phytopathogens fungi including Cytospora sp., Colletotrichum gloeosporioides, Botrytis cinerea, Alternaria solani, and Fusarium solani in vitro. Most of those compounds exhibited remarkable antifungal activities and were superior to the positive control hymexazol.

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

    OpenAIRE

    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...

  3. The significance of ecology in the development of Verticillium chlamydosporium as a biological agent against root-knot nematodes (Meloidogyne spp.).

    OpenAIRE

    Leij, van der, T.

    1992-01-01

    A thorough understanding of the interactions which occur between nematode parasites and nematode pests and the influence of biotic and abiotic factors on these interactions, is essential in the development of biological control agents for nematodes. The aim of this study was to develop a particular isolate of the nematophagous fungus Verticillium chlamydosporium as a biological control agent for root-knot nematodes. The work has gained insight into some of the key factors which govern the eff...

  4. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7

    Science.gov (United States)

    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

  5. Comportamento de variedades paulistas de algodoeiro em solo infestado pelo fungo causador da murcha verticilar Comparative reactions of some cotton varieties to Verticillium wilt in field conditions

    Directory of Open Access Journals (Sweden)

    Edivaldo Cia

    1970-01-01

    Full Text Available Foi estudado o comportamento de variedades paulistas de algodoeiro em terreno infestado pelo fungo causador da "murcha de Verticillium", no Estado de São Paulo. As variedades IAC RM3 e IAC RM4, resistentes à Fusariose, destacaram-se em produção e apresentaram as menores porcentagens de plantas com sintomas externos da doença estudada. Os resultados sugerem que tais variedades constituem também fontes genéticas de resistência à murcha verticilar.A study was made on the reactions of the cotton varieties commonly cultivated in the State of São Paulo (Brazil to Verticillium wilt under field conditions. The effects of the disease were evaluated by determining the percentages of plants showing the symptoms of the Verticilliumwilt and by measuring the yield of each variety. The Fusarium resistant varieties IAC RM3 and IAC RM4 gave the highest yields and the lowest percentages of plants infected by Verticilliumwilt. It is suggested that these varieties might represent sources of resistance to Verticillium wilt.

  6. Major phytopathogens and strains from cocoa (Theobroma cacao L.) are differentiated by MALDI-MS lipid and/or peptide/protein profiles.

    Science.gov (United States)

    Dos Santos, Fábio Neves; Tata, Alessandra; Belaz, Kátia Roberta Anacleto; Magalhães, Dilze Maria Argôlo; Luz, Edna Dora Martins Newman; Eberlin, Marcos Nogueira

    2017-03-01

    Phytopathogens are the main disease agents that promote attack of cocoa plantations in all tropical countries. The similarity of the symptoms caused by different phytopathogens makes the reliable identification of the diverse species a challenge. Correct identification is important in the monitoring and management of these pests. Here we show that matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) in combination with multivariate data analysis is able to rapidly and reliably differentiate cocoa phytopathogens, namely Moniliophthora perniciosa, Phytophthora palmivora, P. capsici, P. citrophthora, P. heveae, Ceratocystis cacaofunesta, C. paradoxa, and C. fimbriata. MALDI-MS reveals unique peptide/protein and lipid profiles which differentiate these phytopathogens at the level of genus, species, and single strain coming from different hosts or cocoa tissues collected in several plantations/places. This fast methodology based on molecular biomarkers is also shown to be sufficiently reproducible and selective and therefore seems to offer a suitable tool to guide the correct application of sanitary defense approaches for infected cocoa plantations. International trading of cocoa plants and products could also be efficiently monitored by MALDI-MS. It could, for instance, prevent the entry of new phytopathogens into a country, e.g., as in the case of Moniliophthora roreri fungus that is present in all cocoa plantations of countries bordering Brazil, but that has not yet attacked Brazilian plantations. Graphical Abstract Secure identification of phytopathogens attacking cocoa plantations has been demonstrated via typical chemical profiles provided by mass spectrometric screening.

  7. Isolation and characterization of a chitinase gene from entomopathogenic fungus Verticillium lecanii Isolamento e caracterização de um gene de quitinase do fungo entomopatogênico Verticillium lecanii

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    Yanping Zhu

    2008-06-01

    Full Text Available Entomopathogenic fungus Verticillium lecanii is a promising whitefly and aphid control agent. Chitinases secreted by this insect pathogen have considerable importance in the biological control of some insect pests. An endochitinase gene Vlchit1 from the fungus was cloned and overexpressed in Escherichia coli. The Vlchit1 gene not only contains an open reading frame (ORF which encodes a protein of 423 amino acids (aa, but also is interrupted by three short introns. A homology modelling of Vlchit1 protein showed that the chitinase Vlchit1 has a (α/β8 TIM barrel structure. Overexpression test and Enzymatic activity assay indicated that the Vlchit1 is a functional enzyme that can hydrolyze the chitin substrate, so the Vlchit1 gene can service as a useful gene source for genetic manipulation leading to strain improvement of entomopathogenic fungi or constructing new transgenic plants with resistance to various fungal and insects pests.O fungo entomopatogênico Verticillium lecanii é um agente promissor no controle da mosca-branca e do pulgão. As quitinases secretadas por esse patógeno de insetos têm uma grande importância no controle biológico de doenças causadas por insetos. Um gene de endoquitinase Vlchit1 desse fungo foi clonado e expresso em Escherichia coli. O gene Vlchit contém não apenas um ORF que codifica uma proteína de 423 aminoácidos, mas também é interrompido por três pequenos introns. A modelagem de homologia da proteína Vlchit1indicou que a quitinase Vlchit1 tem uma estrutura (α/β 8 TIM barrel. Testes de expressão e de atividade enzimática indicaram que Vlchit1 é uma enzima funcional que hidroliza quitina, portanto o gene Vlchit pode ser um gene útil para manipulação genética para melhoramento de cepas de fungos entomopatogênicos ou para a construção de novas plantas transgênicas com resistência a várias doenças causadas por fungos e insetos.

  8. Cytoplasmic- and extracellular-proteome analysis of Diplodia seriata: a phytopathogenic fungus involved in grapevine decline

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    Cobos Rebeca

    2010-09-01

    Full Text Available Abstract Background The phytopathogenic fungus Diplodia seriata, whose genome remains unsequenced, produces severe infections in fruit trees (fruit blight and grapevines. In this crop is recognized as one of the most prominent pathogens involved in grapevine trunk disease (or grapevine decline. This pathology can result in the death of adult plants and therefore it produces severe economical losses all around the world. To date no genes or proteins have been characterized in D. seriata that are involved in the pathogenicity process. In an effort to help identify potential gene products associated with pathogenicity and to gain a better understanding of the biology of D. seriata, we initiated a proteome-level study of the fungal mycelia and secretome. Results Intracellular and secreted proteins from D. seriata collected from liquid cultures were separated using two-dimensional gel electrophoresis. About 550 cytoplasmic proteins were reproducibly present in 3 independent extractions, being 53 identified by peptide mass fingerprinting and tandem mass spectrometry. The secretome analysis showed 75 secreted proteins reproducibly present in 3 biological replicates, being 16 identified. Several of the proteins had been previously identified as virulence factors in other fungal strains, although their contribution to pathogenicity in D. seriata remained to be analyzed. When D. seriata was grown in a medium supplemented with carboxymethylcellulose, 3 proteins were up-regulated and 30 down-regulated. Within the up-regulated proteins, two were identified as alcohol dehydrogenase and mitochondrial peroxyrredoxin-1, suggesting that they could play a significant role in the pathogenicity process. As for the 30 down-regulated proteins, 9 were identified being several of them involved in carbohydrate metabolism. Conclusions This study is the first report on proteomics on D. seriata. The proteomic data obtained will be important to understand the pathogenicity

  9. Detection of Verticillium wilt of olive trees and downy mildew of opium poppy using hyperspectral and thermal UAV imagery

    Science.gov (United States)

    Calderón Madrid, Rocío; Navas Cortés, Juan Antonio; Montes Borrego, Miguel; Landa del Castillo, Blanca Beatriz; Lucena León, Carlos; Jesús Zarco Tejada, Pablo

    2014-05-01

    The present study explored the use of high-resolution thermal, multispectral and hyperspectral imagery as indicators of the infections caused by Verticillium wilt (VW) in olive trees and downy mildew (DM) in opium poppy fields. VW, caused by the soil-borne fungus Verticillium dahliae, and DM, caused by the biotrophic obligate oomycete Peronospora arborescens, are the most economically limiting diseases of olive trees and opium poppy, respectively, worldwide. V. dahliae infects the plant by the roots and colonizes its vascular system, blocking water flow and eventually inducing water stress. P. arborescens colonizes the mesophyll, appearing the first symptoms as small chlorotic leaf lesions, which can evolve to curled and thickened tissues and systemic infections that become deformed and necrotic as the disease develops. The work conducted to detect VW and DM infection consisted on the acquisition of time series of airborne thermal, multispectral and hyperspectral imagery using 2-m and 5-m wingspan electric Unmanned Aerial Vehicles (UAVs) in spring and summer of three consecutive years (2009 to 2011) for VW detection and on three dates in spring of 2009 for DM detection. Two 7-ha commercial olive orchards naturally infected with V. dahliae and two opium poppy field plots artificially infected by P. arborescens were flown. Concurrently to the airborne campaigns, olive orchards and opium poppy fields were assessed "in situ" to assess actual VW severity and DM incidence. Furthermore, field measurements were conducted at leaf and crown level. The field results related to VW detection showed a significant increase in crown temperature (Tc) minus air temperature (Ta) and a decrease in leaf stomatal conductance (G) as VW severity increased. This reduction in G was associated with a significant increase in the Photochemical Reflectance Index (PRI570) and a decrease in chlorophyll fluorescence. DM asymptomatic leaves showed significantly higher NDVI and lower green/red index

  10. Liquid Swine Manure Can Kill Verticillium dahliae Microsclerotia in Soil by Volatile Fatty Acid, Nitrous Acid, and Ammonia Toxicity.

    Science.gov (United States)

    Conn, Kenneth L; Tenuta, Mario; Lazarovits, George

    2005-01-01

    ABSTRACT In previous studies, liquid swine manure (LSM) was sometimes shown to reduce Verticillium wilt of potato caused by Verticillium dahliae. We also observed that microsclerotia of this fungus died within 1 day, or between 3 and 6 weeks, after addition of LSM to some acid soils and within 1 week in some alkaline soils. In this study, we demonstrated that a volatile fatty acid (VFA) mixture with an identical concentration of VFAs as that found in an effective LSM reduced germination in an acid soil (pH 5.1) to the same extent as the LSM after 1 day of exposure. Germination was reduced by 45, 75, and 90% in the 10, 20, and 40% ([wt/wt] soil moisture) treatments, respectively, with the latter being equivalent to an application of 80 hl/ha. Addition to this acid soil of 19 LSMs (30% [wt/wt] soil moisture) collected from different producers resulted in complete kill of microsclerotia with 12 manures. Effective manures had a total concentration of nonionized forms of VFAs in soil solution of 2.7 mM or higher. In some acid soils (pH 5.8), addition of LSM (40% [wt/wt] soil moisture) did not kill microsclerotia until 3 to 6 weeks later. Here, a reduction in viability of microsclerotia was attributed to the accumulation of 0.06 mM nitrous acid in the soil solution at 4 weeks. When an LSM was added (40% [wt/wt] soil moisture) to an alkaline soil (pH 7.9) where VFAs are not toxic, microsclerotia germination was reduced by 80% after 1 week. Here the pH increased to 8.9 and the concentration of ammonia reached 30 mM in the soil solution. An ammonium chloride solution having an equivalent concentration of ammonium as the manure was shown to have the same spectrum of toxicity as the manure in assays ranging from pH 7 to 9, both in solutions and above the solutions. At pH 9, the concentration of ammonia reached 18 mM and 100% mortality of microsclerotia occurred. Thus, in acid soils, LSM can kill microsclerotia of V. dahliae by VFA and/or nitrous acid toxicity and in alkaline

  11. Preliminary Study on Screening and Antagonistic Mechanism of Indicator Microbe for Wilting Toxin from Verticillium dahliae%大丽轮枝菌致萎毒力指示菌的筛选及拮抗作用初探

    Institute of Scientific and Technical Information of China (English)

    王世英; 姜军坡; 李术娜; 朱宝成; 薛庆林

    2010-01-01

    为了筛选可指示Verticillium dahliae致萎毒素毒力大小的指示菌.并对致萎毒素抑制指示菌生长的机理进行初步探讨,采用指示菌平板打孔法,对黑曲霉(Aspergillus niger)BF-3758、枯草芽孢杆菌(Bacillus subtilis)、金黄色葡萄球(Staphylococcus aurcus)、啤酒酵母(Saccharomyces cerevisiae)等12种具有微生物种属代表性的菌株进行筛选:采用拮抗试验、致萎试验结合显微观察的方法,初步探索了Verticillium dahliae致萎毒素对指示菌生长的抑制作用.结果表明,Verticillium dahliae粗毒素对Aspergillus niger BF-3758的生长具有较强的抑制作用,且该抑制作用和粗毒素对离体叶片的致萎作用具有一致性,并存在一定的量效关系,Aspergillus niger BF-3758菌株可以作为Verticillium dahliae致萎毒素的毒力指示菌;Verticillium dahliae粗毒素可抑制Aspergillus niger BF-3758的孢子萌发和菌丝生长,对成熟菌丝也有破坏作用.上述研究可为评价Verticillium dahliae的致病力提供一种操作简便、耗时短、费用低的生物学方法.

  12. Trichoderma species mediated differential tolerance against biotic stress of phytopathogens in Cicer arietinum L.

    Science.gov (United States)

    Saxena, Amrita; Raghuwanshi, Richa; Singh, Harikesh Bahadur

    2015-02-01

    Trichoderma spp. have been reported to aid in imparting biotic as well as abiotic tolerance to plants. However, there are only few reports unfolding the differential ability of separate species of Trichoderma genera generally exploited for their biocontrol potential in this framework. A study was undertaken to evaluate the biocontrol potential of different Trichoderma species namely T. harzianum, T. asperellum, T. koningiopsis, T. longibrachiatum, and T. aureoviride as identified in the group of indigenous isolates from the agricultural soils of Eastern Uttar Pradesh, India. Their biocontrol potential against three major soilborne phytopathogens, i.e., Sclerotium rolfsii, Sclerotinia sclerotiorum, and Colletotrichum capsici was confirmed by dual culture plate technique. Efficient mycoparasitic ability was further assessed in all the isolates in relation to chitinase, β-1,3 glucanase, pectinase, lipase, amylase, and cellulase production while equally consistent results were obtained for their probable phosphate solubilization and indole acetic acid (IAA) production abilities. The selected isolates were further subjected to test their ability to promote plant growth, to reduce disease incidence and to tolerate biotic stress in terms of lignification pattern against S. rolfsii in chickpea plants. Among the identified Trichoderma species, excellent results were observed for T. harzianum and T. koningiopsis indicating better biocontrol potential of these species in the group and thus exhibiting perspective for their commercial exploitation.

  13. Efeito de herbicidas sobre agentes fitopatogênicos = Effect of herbicides on phytopathogenic agents

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    Daniel Dias Rosa

    2010-07-01

    Full Text Available Na agricultura moderna, diversas tecnologias auxiliam no aumento daprodutividade, sendo o herbicida uma delas, mas existem consequências atreladas ao seu uso, como os diversos efeitos sobre organismos não alvos. Neste trabalho, objetivou-se verificar esses efeitos sobre agentes fitopatogênicos, assim como avaliar o efeito do herbicida glyphosate sobre diversas doenças, em plantas de soja transgênicas.Verificou-se forte ação fungicida com o uso do herbicida glyphosate, assim como os outros avaliados “in vitro”, sobre os fungos testados, e os mesmos resultados foram observados nas plantas em condição de campo.In modern agriculture, several technologies have helped increase productivity, and herbicide is one of them. However, there are consequences linked to its use, such as the various effects on non-target organisms. The purpose of this work was to verify these effects on phytopathogenic agents, as well as assess the effect of glyphosate on diseases in transgenic soybean. There was a strong fungicide action using glyphosate herbicide as well as with the others evaluated in vitro regarding fungi tested. The same results were observed in plants in field conditions.

  14. Antagonism and Molecular Identification of an Antibiotic Bacterium BS04 Against Phytopathogenic Fungi and Bacteria

    Institute of Scientific and Technical Information of China (English)

    Xie Jing(谢晶); Ge Shaorong; Tao Yong; Gao Ping; Liu Kun; Liu Shigui

    2004-01-01

    Through a modified agar well diffusion assay, antagonism of bacterium BS04 is tested. The data show that BS04 has antibiotic activity against phytopathogenic fungi and bacteria, including Phoma wasabiae Yokogi, Cochlibolus Heterostrophu, Exserohilum Turcicum, Curuvularia Lunata (Walk) Boed, Thantephorus cucumris, Fusarium graminearum, Xanthomonas axonopodis pv. Citri (Hasse) Dye and Xanthomonas zingiberi (Uyeda) Savulescu. The products of bacterium BS04 can endure the treatment of a wide range of pH, and maintain the antibiotic activity after treatment of 100℃ for 30 min. The result suggests that bacterium BS04 has the potential as a promising biocontrol agent. In order to determine the taxonomic placement, the molecular identification of BS04 is performed. The comparative analysis of 16s rDNA sequences indicates that the 16s rDNA sequence of BS04 is highly homologous with sequences of typical Paenibacillus bacteria from the RPD library (from 92% to 99%). And the constructed phylogenetic tree by using maximum-likelihood method with Bootstrap Trial 1000 proves that BS04 is subjected to Paenibacillus polymyxa.

  15. Differences in stability of seed-associated microbial assemblages in response to invasion by phytopathogenic microorganisms

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    Samir Rezki

    2016-04-01

    Full Text Available Seeds are involved in the vertical transmission of microorganisms from one plant generation to another and consequently act as reservoirs for the plant microbiota. However, little is known about the structure of seed-associated microbial assemblages and the regulators of assemblage structure. In this work, we have assessed the response of seed-associated microbial assemblages of Raphanus sativus to invading phytopathogenic agents, the bacterial strain Xanthomonas campestris pv. campestris (Xcc 8004 and the fungal strain Alternaria brassicicola Abra43. According to the indicators of bacterial (16S rRNA gene and gyrB sequences and fungal (ITS1 diversity employed in this study, seed transmission of the bacterial strain Xcc 8004 did not change the overall composition of resident microbial assemblages. In contrast seed transmission of Abra43 strongly modified the richness and structure of fungal assemblages without affecting bacterial assemblages. The sensitivity of seed-associated fungal assemblage to Abra43 is mostly related to changes in relative abundance of closely related fungal species that belong to the Alternaria genus. Variation in stability of the seed microbiota in response to Xcc and Abra43 invasions could be explained by differences in seed transmission pathways employed by these micro-organisms, which ultimately results in divergence in spatio-temporal colonization of the seed habitat.

  16. Differences in stability of seed-associated microbial assemblages in response to invasion by phytopathogenic microorganisms.

    Science.gov (United States)

    Rezki, Samir; Campion, Claire; Iacomi-Vasilescu, Beatrice; Preveaux, Anne; Toualbia, Youness; Bonneau, Sophie; Briand, Martial; Laurent, Emmanuelle; Hunault, Gilles; Simoneau, Philippe; Jacques, Marie-Agnès; Barret, Matthieu

    2016-01-01

    Seeds are involved in the vertical transmission of microorganisms from one plant generation to another and consequently act as reservoirs for the plant microbiota. However, little is known about the structure of seed-associated microbial assemblages and the regulators of assemblage structure. In this work, we have assessed the response of seed-associated microbial assemblages of Raphanus sativus to invading phytopathogenic agents, the bacterial strain Xanthomonas campestris pv. campestris (Xcc) 8004 and the fungal strain Alternaria brassicicola Abra43. According to the indicators of bacterial (16S rRNA gene and gyrB sequences) and fungal (ITS1) diversity employed in this study, seed transmission of the bacterial strain Xcc 8004 did not change the overall composition of resident microbial assemblages. In contrast seed transmission of Abra43 strongly modified the richness and structure of fungal assemblages without affecting bacterial assemblages. The sensitivity of seed-associated fungal assemblage to Abra43 is mostly related to changes in relative abundance of closely related fungal species that belong to the Alternaria genus. Variation in stability of the seed microbiota in response to Xcc and Abra43 invasions could be explained by differences in seed transmission pathways employed by these micro-organisms, which ultimately results in divergence in spatio-temporal colonization of the seed habitat.

  17. Transmission of Fusarium boothii mycovirus via protoplast fusion causes hypovirulence in other phytopathogenic fungi.

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    Kyung-Mi Lee

    Full Text Available There is increasing concern regarding the use of fungicides to control plant diseases, whereby interest has increased in the biological control of phytopathogenic fungi by the application of hypovirulent mycoviruses as a possible alternative to fungicides. Transmission of hypovirulence-associated double-stranded RNA (dsRNA viruses between mycelia, however, is prevented by the vegetative incompatibility barrier that often exists between different species or strains of filamentous fungi. We determined whether protoplast fusion could be used to transmit FgV1-DK21 virus, which is associated with hypovirulence on F. boothii (formerly F. graminearum strain DK21, to F. graminearum, F. asiaticum, F. oxysporum f. sp. lycopersici, and Cryphonectria parasitica. Relative to virus-free strains, the FgV1-DK21 recipient strains had reduced growth rates, altered pigmentation, and reduced virulence. These results indicate that protoplast fusion can be used to introduce FgV1-DK21 dsRNA into other Fusarium species and into C. parasitica and that FgV1-DK21 can be used as a hypovirulence factor and thus as a biological control agent.

  18. Emerging phytopathogen Macrophomina phaseolina: biology, economic importance and current diagnostic trends.

    Science.gov (United States)

    Kaur, Surinder; Dhillon, Gurpreet Singh; Brar, Satinder Kaur; Vallad, Gary Edward; Chand, Ramesh; Chauhan, Vijay Bahadur

    2012-05-01

    Macrophomina phaseolina (Tassi) Goid. is an important phytopathogenic fungus, infecting a large number of plant species and surviving for up to 15 years in the soil as a saprophyte. Although considerable research related to the biology and ecology of Macrophomina has been conducted, it continues to cause huge economic losses in many crops. Research is needed to improve the identification and characterization of genetic variability within their epidemiological and pathological niches. Better understanding of the variability within the pathogen population for traits that influence fitness and soil survival will certainly lead to improved management strategies for Macrophomina. In this context, the present review discusses various biological aspects and distribution of M. phaseolina throughout the world and their importance to different plant species. Accurate identification of the fungus has been aided with the use of nucleic acid-based molecular techniques. The development of PCR-based methods for identification and detection of M. phaseolina are highly sensitive and specific. Early diagnosis and accurate detection of pathogens is an essential step in plant disease management as well as quarantine. The progress in the development of various molecular tools used for the detection, identification and characterization of Macrophomina isolates were also discussed.

  19. Isolation of Bacteria with Antifungal Activity against the Phytopathogenic Fungi Stenocarpella maydis and Stenocarpella macrospora

    Science.gov (United States)

    Petatán-Sagahón, Iván; Anducho-Reyes, Miguel Angel; Silva-Rojas, Hilda Victoria; Arana-Cuenca, Ainhoa; Tellez-Jurado, Alejandro; Cárdenas-Álvarez, Isabel Oyuki; Mercado-Flores, Yuridia

    2011-01-01

    Stenocarpella maydis and Stenocarpella macrospora are the causal agents of ear rot in corn, which is one of the most destructive diseases in this crop worldwide. These fungi are important mycotoxin producers that cause different pathologies in farmed animals and represent an important risk for humans. In this work, 160 strains were isolated from soil of corn crops of which 10 showed antifungal activity against these phytopathogens, which, were identified as: Bacillus subtilis, Pseudomonas spp., Pseudomonas fluorescens, and Pantoea agglomerans by sequencing of 16S rRNA gene and the phylogenetic analysis. From cultures of each strain, extracellular filtrates were obtained and assayed to determine antifungal activity. The best filtrates were obtained in the stationary phase of B. subtilis cultures that were stable to the temperature and extreme pH values; in addition they did not show a cytotoxicity effect against brine shrimp and inhibited germination of conidia. The bacteria described in this work have the potential to be used in the control of white ear rot disease. PMID:22016606

  20. In Vitro Antifungal Activity of Burkholderia gladioli pv. agaricicola against Some Phytopathogenic Fungi

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    Hazem S. Elshafie

    2012-12-01

    Full Text Available The trend to search novel microbial natural biocides has recently been increasing in order to avoid the environmental pollution from use of synthetic pesticides. Among these novel natural biocides are the bioactive secondary metabolites of Burkholderia gladioli pv. agaricicola (Bga. The aim of this study is to determine antifungal activity of Bga strains against some phytopathogenic fungi. The fungicidal tests were carried out using cultures and cell-free culture filtrates against Botrytis cinerea, Aspergillus flavus, Aspergillus niger, Penicillium digitatum, Penicillium expansum, Sclerotinia sclerotiorum and Phytophthora cactorum. Results demonstrated that all tested strains exert antifungal activity against all studied fungi by producing diffusible metabolites which are correlated with their ability to produce extracellular hydrolytic enzymes. All strains significantly reduced the growth of studied fungi and the bacterial cells were more bioactive than bacterial filtrates. All tested Bulkholderia strains produced volatile organic compounds (VOCs, which inhibited the fungal growth and reduced the growth rate of Fusarium oxysporum and Rhizoctonia solani. GC/MS analysis of VOCs emitted by strain Bga 11096 indicated the presence of a compound that was identified as 1-methyl-4-(1-methylethenyl-cyclohexene, a liquid hydrocarbon classified as cyclic terpene. This compound could be responsible for the antifungal activity, which is also in agreement with the work of other authors.

  1. The effects of some polyamine biosynthetic inhibitors on growth and morphology of phytopathogenic fungi.

    Science.gov (United States)

    Rajam, M V; Galston, A W

    1985-01-01

    We have studied the effects of two polyamine biosynthetic inhibitors, alpha-difluoromethylornithine (DFMO) and alpha-difluoromethylarginine (DFMA), and of polyamines (PAs), alone and in combination, on mycelial growth and morphology of four phytopathogenic fungi: Botrytis sp, B. cinerea, Rhizoctonia solani and Monilinia fructicola. The inhibitors were added to a Czapek agar medium to get final concentrations of 0.1, 0.5 and 1.0 mM. DFMO and DFMA, suicide inhibitors of ornithine decarboxylase (ODC) and arginine decarboxylase (ADC) respectively, inhibited mycelial growth strongly; the effect was generally more pronounced with DFMA than with DFMO, but each fungus had its own response pattern. The addition of the PAs putrescine (Put) and spermidine (Spd) to the culture medium resulted in a promotion of growth. In Botrytis sp and Monilinia fructicola exposed to inhibitors plus PAs, mycelial growth was actually increased above control values. Mycelial morphology was altered and cell size dramatically reduced in plates containing inhibitors alone, whereas with PAs alone, or in combination with inhibitors, morphology was normal, but cell length and diameters increased considerably. These results suggest that PAs are essential for growth in fungal mycelia. The inhibition caused by DFMA may be due to its arginase-mediated conversion to DFMO.

  2. Propiconazole inhibits the sterol 14α-demethylase in Glomus irregulare like in phytopathogenic fungi.

    Science.gov (United States)

    Calonne, Maryline; Sahraoui, Anissa Lounès-Hadj; Campagnac, Estelle; Debiane, Djouher; Laruelle, Frédéric; Grandmougin-Ferjani, Anne; Fontaine, Joël

    2012-04-01

    The increasing concentrations impact (0.02, 0.2 and 2 mg L(-1)) of a Sterol Biosynthesis Inhibitor (SBI) fungicide, propiconazole, was evaluated on development and sterol metabolism of two non-target organisms: mycorrhizal or non-mycorrhizal transformed chicory roots and the arbuscular mycorrhizal fungus (AMF) Glomus irregulare using monoxenic cultures. In this work, we provide the first evidence of a direct impact of propiconazole on the AMF by disturbing its sterol metabolism. A significant decrease in end-products sterols contents (24-methylcholesterol and in 24-ethylcholesterol) was observed concomitantly to a 24-methylenedihydrolanosterol accumulation indicating the inhibition of a key enzyme in sterol biosynthesis pathway, the sterol 14α-demethylase like in phytopathogenic fungi. A decrease in end-product sterol contents in propiconazole-treated roots was also observed suggesting a slowing down of the sterol metabolism in plant. Taken together, our findings suggest that the inhibition of the both AM symbiotic partners development by propiconazole results from their sterol metabolism alterations.

  3. Volatile organic compounds produced by the phytopathogenic bacterium Xanthomonas campestris pv. vesicatoria 85-10

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    Teresa Weise

    2012-04-01

    Full Text Available Xanthomonas campestris is a phytopathogenic bacterium and causes many diseases of agricultural relevance. Volatiles were shown to be important in inter- and intraorganismic attraction and defense reactions. Recently it became apparent that also bacteria emit a plethora of volatiles, which influence other organisms such as invertebrates, plants and fungi. As a first step to study volatile-based bacterial–plant interactions, the emission profile of Xanthomonas c. pv. vesicatoria 85-10 was determined by using GC/MS and PTR–MS techniques. More than 50 compounds were emitted by this species, the majority comprising ketones and methylketones. The structure of the dominant compound, 10-methylundecan-2-one, was assigned on the basis of its analytical data, obtained by GC/MS and verified by comparison of these data with those of a synthetic reference sample. Application of commercially available decan-2-one, undecan-2-one, dodecan-2-one, and the newly synthesized 10-methylundecan-2-one in bi-partite Petri dish bioassays revealed growth promotions in low quantities (0.01 to 10 μmol, whereas decan-2-one at 100 μmol caused growth inhibitions of the fungus Rhizoctonia solani. Volatile emission profiles of the bacteria were different for growth on media (nutrient broth with or without glucose.

  4. Volatile organic compounds produced by the phytopathogenic bacterium Xanthomonas campestris pv. vesicatoria 85-10.

    Science.gov (United States)

    Weise, Teresa; Kai, Marco; Gummesson, Anja; Troeger, Armin; von Reuß, Stephan; Piepenborn, Silvia; Kosterka, Francine; Sklorz, Martin; Zimmermann, Ralf; Francke, Wittko; Piechulla, Birgit

    2012-01-01

    Xanthomonas campestris is a phytopathogenic bacterium and causes many diseases of agricultural relevance. Volatiles were shown to be important in inter- and intraorganismic attraction and defense reactions. Recently it became apparent that also bacteria emit a plethora of volatiles, which influence other organisms such as invertebrates, plants and fungi. As a first step to study volatile-based bacterial-plant interactions, the emission profile of Xanthomonas c. pv. vesicatoria 85-10 was determined by using GC/MS and PTR-MS techniques. More than 50 compounds were emitted by this species, the majority comprising ketones and methylketones. The structure of the dominant compound, 10-methylundecan-2-one, was assigned on the basis of its analytical data, obtained by GC/MS and verified by comparison of these data with those of a synthetic reference sample. Application of commercially available decan-2-one, undecan-2-one, dodecan-2-one, and the newly synthesized 10-methylundecan-2-one in bi-partite Petri dish bioassays revealed growth promotions in low quantities (0.01 to 10 μmol), whereas decan-2-one at 100 μmol caused growth inhibitions of the fungus Rhizoctonia solani. Volatile emission profiles of the bacteria were different for growth on media (nutrient broth) with or without glucose.

  5. ANTIFUNGAL EFFICACY OF AQUEOUS EXTRACTS OF NEEM CAKE, KARANJ CAKE AND VERMICOMPOST AGAINST SOME PHYTOPATHOGENIC FUNGI

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    JAIPAL SINGH CHOUDHARY

    2013-01-01

    Full Text Available Several agro-based waste and byproducts are known and cited to play an important role in the management ofplant diseases in ancient texts. They act directly or indirectly on plant pathogens to inhibit the growth andmultiplication or by inducing resistance in crop plants. In the present experiments, aqueous extracts of neemcake, karanj cake and vermicompost were tested against some important phytopathogenic fungi viz.,Helminthosporium pennisetti, Curvularia lunata and Colletotrichum gloeosporioides f. sp. mangiferae for theirantifungal activities. H. pennisetti was found to be most sensitive one followed by C. gloeosporioides f. sp.mangiferae and C. lunata against all the tested drugs. Against C. gloeosporioides f. sp. mangiferae, aqueous extractof karanj cake was most effective where GI50 was found to be 0.41% drug concentration followed by neem cake(0.46% and vermicompost (0.86%. In case of C. lunata and H. pennisetti, neem cake extract was most effectivewith GI50 value of 0.27% and 0.11% respectively. The GI50 values noted for C. lunata with extracts of karanj cake(0.70%, vermicompost (0.88% and for H. pennisetti were (0.20% and (0.22% respectively.

  6. Synthesis of chitosan based nanoparticles and their in vitro evaluation against phytopathogenic fungi.

    Science.gov (United States)

    Saharan, Vinod; Mehrotra, Akanksha; Khatik, Rajesh; Rawal, Pokhar; Sharma, S S; Pal, Ajay

    2013-11-01

    The main aim of present study was to prepare chitosan, chitosan-saponin and Cu-chitosan nanoparticles to evaluate their in vitro antifungal activities. Various nanoparticles were prepared using ionic gelation method by interaction of chitosan, sodium tripolyphosphate, saponin and Cu ions. Their particle size, polydispersity index, zeta potential and structures were confirmed by DLS, FTIR, TEM and SEM. The antifungal properties of nanoparticles against phytopathogenic fungi namely Alternaria alternata, Macrophomina phaseolina and Rhizoctonia solani were investigated at various concentrations ranging from 0.001 to 0.1%. Among the various formulations of nanoparticles, Cu-chitosan nanoparticles were found most effective at 0.1% concentration and showed 89.5, 63.0 and 60.1% growth inhibition of A. alternata, M. phaseolina and R. solani, respectively in in vitro model. At the same concentration, Cu-chitosan nanoparticles also showed maximum of 87.4% inhibition rate of spore germination of A. alternata. Chitosan nanoparticles showed the maximum growth inhibitory effects (87.6%) on in vitro mycelial growth of M. phaseolina at 0.1% concentration. From our study it is evident that chitosan based nanoparticles particularly chitosan and Cu-chitosan nanoparticles have tremendous potential for further field screening towards crop protection.

  7. Production of lipopeptides among Bacillus strains showing growth inhibition of phytopathogenic fungi.

    Science.gov (United States)

    Velho, R V; Medina, L F C; Segalin, J; Brandelli, A

    2011-07-01

    The biological activity and the presence of genes sfp and ituD (surfactin and iturin A) among Bacillus strains isolated from the Amazon basin were determined. Bacillus spp. were tested for hemolytic activity and inhibition of fungal growth by agar plate assays in parallel with PCR for identification of sfp and ituD genes. All strains tested produced surface-active compounds, giving evidence by lysis of erythrocytes and emulsifying activity on mineral oil and soybean oil. These strains of Bacillus caused growth inhibition of several phytopathogenic fungi, including Fusarium spp., Aspergillus spp., and Bipolaris sorokiniana. The presence of genes ituD and sfp was confirmed by PCR and sequence analysis. The only exception was Bacillus sp. P34 that lacks sfp gene. Lipopeptides were isolated from culture supernatants and analyzed by mass spectrometry. Characteristic m/z peaks for surfactin and iturin were observed, and some strains also produced fengycin and bacillomycin. The remarkable antifungal activity showed by the strains could be associated with the co-production of three or more lipopeptide antibiotics. Screening for novel bacteria producing useful biosurfactants or biocontrol agents for agriculture is a topic of greatest importance to eliminate chemical pollutants.

  8. Enhancement of the Antibacterial Activity of Silver Nanoparticles against Phytopathogenic Bacterium Ralstonia solanacearum by Stabilization

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    Juanni Chen

    2016-01-01

    Full Text Available In this paper, the enhanced antibacterial activity of silver nanoparticles (AgNPs against the phytopathogenic bacterium Ralstonia solanacearum after stabilization using selected surfactants (SDS, SDBS, TX-100, and Tween 80 was examined, in comparison with silver ion. Tween 80 was found to be the most preferable stabilizer of AgNPs due to the beneficial synergistic effects of the AgNPs and surfactant. However, all the surfactants nearly had no effects on the antibacterial activity of Ag+. In vitro, Tween 80-stabilized AgNPs showed the highest bactericidal activity against R. solanacearum. Further measurements using TEM, fluorescence microscopy, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE revealed that though Ag+ and Tween 80-Ag+ induced high toxicity, Tween 80-stabilized AgNPs displayed most severe damage when in direct contact with cells, causing mechanistic injury to the cell membrane and strongly modifying and destructing the cellular proteins. Meanwhile, in vivo, the pot experiments data indicated that the control efficiency of Tween 80-stabilized AgNPs on tobacco bacterial wilt was 96.71%, 90.11%, and 84.21%, at 7 days, 14 days, and 21 days, respectively. Based on the results evidencing their advantageous low dosage requirements and strong antimicrobial activity, Tween 80-stabilized AgNPs are a promising antibacterial agent for use in alternative crop disease control approaches.

  9. Isolation of Bacteria with Antifungal Activity against the Phytopathogenic Fungi Stenocarpella maydis and Stenocarpella macrospora

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    Yuridia Mercado-Flores

    2011-08-01

    Full Text Available Stenocarpella maydis and Stenocarpella macrospora are the causal agents of ear rot in corn, which is one of the most destructive diseases in this crop worldwide. These fungi are important mycotoxin producers that cause different pathologies in farmed animals and represent an important risk for humans. In this work, 160 strains were isolated from soil of corn crops of which 10 showed antifungal activity against these phytopathogens, which, were identified as: Bacillus subtilis, Pseudomonas spp., Pseudomonas fluorescens, and Pantoea agglomerans by sequencing of 16S rRNA gene and the phylogenetic analysis. From cultures of each strain, extracellular filtrates were obtained and assayed to determine antifungal activity. The best filtrates were obtained in the stationary phase of B. subtilis cultures that were stable to the temperature and extreme pH values; in addition they did not show a cytotoxicity effect against brine shrimp and inhibited germination of conidia. The bacteria described in this work have the potential to be used in the control of white ear rot disease.

  10. Isolation of bacteria with antifungal activity against the phytopathogenic fungi Stenocarpella maydis and Stenocarpella macrospora.

    Science.gov (United States)

    Petatán-Sagahón, Iván; Anducho-Reyes, Miguel Angel; Silva-Rojas, Hilda Victoria; Arana-Cuenca, Ainhoa; Tellez-Jurado, Alejandro; Cárdenas-Álvarez, Isabel Oyuki; Mercado-Flores, Yuridia

    2011-01-01

    Stenocarpella maydis and Stenocarpella macrospora are the causal agents of ear rot in corn, which is one of the most destructive diseases in this crop worldwide. These fungi are important mycotoxin producers that cause different pathologies in farmed animals and represent an important risk for humans. In this work, 160 strains were isolated from soil of corn crops of which 10 showed antifungal activity against these phytopathogens, which, were identified as: Bacillus subtilis, Pseudomonas spp., Pseudomonas fluorescens, and Pantoea agglomerans by sequencing of 16S rRNA gene and the phylogenetic analysis. From cultures of each strain, extracellular filtrates were obtained and assayed to determine antifungal activity. The best filtrates were obtained in the stationary phase of B. subtilis cultures that were stable to the temperature and extreme pH values; in addition they did not show a cytotoxicity effect against brine shrimp and inhibited germination of conidia. The bacteria described in this work have the potential to be used in the control of white ear rot disease.

  11. Synthesis and characterization of silver nanoparticles: effect on phytopathogen Colletotrichum gloesporioides

    Energy Technology Data Exchange (ETDEWEB)

    Aguilar-Mendez, Miguel A., E-mail: maguilarme@ipn.mx; San Martin-Martinez, Eduardo; Ortega-Arroyo, Lesli [Instituto Politecnico Nacional, Centro de Investigacion en Ciencia Aplicada y Tecnologia Avanzada (Mexico); Cobian-Portillo, Georgina [Instituto Politecnico Nacional, Centro Interdisciplinario de Investigacion para el Desarrollo Integral Regional (Mexico); Sanchez-Espindola, Esther [Instituto Politecnico Nacional, Escuela Nacional de Ciencias Biologicas, Prolongacion Manuel M. Carpio s/n, esq. Plan de Ayala (Mexico)

    2011-06-15

    Colloidal silver nanoparticles were synthesized by reducing silver nitrate solutions with glucose, in the presence of gelatin as capping agent. The obtained nanoparticles were characterized by means of UV-Vis spectroscopy, transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) spectroscopy. The response surface methodology (RSM) was also used to determine the influence of the variables on the size of the nanoparticles. The antifungal activity of the silver nanoparticles was evaluated on the phytopathogen Colletotrichum gloesporioides, which causes anthracnose in a wide range of fruits. The UV-Vis spectra indicated the formation of silver nanoparticles preferably spherical and of relatively small size (<20 nm). The above-mentioned was confirmed by TEM, observing a size distribution of 5-24 nm. According to RSM the synthesis variables influenced on the size of the silver nanoparticles. By means of FTIR spectroscopy it was determined that gelatin, through their amide and hydroxyl groups, interacts with nanoparticles preventing their agglomeration. The growth of C. gloesporioides in the presence of silver nanoparticles was significantly delayed in a dose dependent manner.

  12. Antagonism of Bacillus spp. isolated from marine biofilms against terrestrial phytopathogenic fungi.

    Science.gov (United States)

    Ortega-Morales, B O; Ortega-Morales, F N; Lara-Reyna, J; De la Rosa-García, S C; Martínez-Hernández, A; Montero-M, Jorge

    2009-01-01

    We aimed at determining the antagonistic behavior of bacteria derived from marine biofilms against terrestrial phytopathogenic fungi. Some bacteria closely related to Bacillus mojavensis (three isolates) and Bacillus firmus (one isolate) displayed antagonistic activity against Colletotrichum gloeosporioides ATCC 42374, selected as first screen organism. The four isolates were further quantitatively tested against C. gloeosporioides, Colletotrichum fragariae, and Fusarium oxysporum on two culture media, potato dextrose agar (PDA) and a marine medium-based agar [yeast extract agar (YEA)] at different times of growth of the antagonists (early, co-inoculation with the pathogen and late). Overall antagonistic assays showed differential susceptibility among the pathogens as a function of the type of culture media and time of colonization (P Bacillus sp. MC3B-22 displayed a greater antagonistic effect than the commercial biocontrol strain Bacillus subtilis G03 (Kodiak). Further incubation studies and scanning electronic microscopy revealed that Bacillus sp. MC3B-22 was able to colonize, multiply, and inhibit C. gloeosporioides ATCC 42374 when tested in a mango leaf assay, showing its potential for fungal biocontrol. Additional studies are required to definitively identify the active isolates and to determine their mode of antifungal action, safety, and biocompatibility.

  13. The Innate Immune Signaling System as a Regulator of Disease Resistance and Induced Systemic Resistance Activity Against Verticillium dahliae.

    Science.gov (United States)

    Gkizi, Danai; Lehmann, Silke; L'Haridon, Floriane; Serrano, Mario; Paplomatas, Epaminondas J; Métraux, Jean-Pierre; Tjamos, Sotirios E

    2016-04-01

    In the last decades, the plant innate immune responses against pathogens have been extensively studied, while biocontrol interactions between soilborne fungal pathogens and their hosts have received much less attention. Treatment of Arabidopsis thaliana with the nonpathogenic bacterium Paenibacillus alvei K165 was shown previously to protect against Verticillium dahliae by triggering induced systemic resistance (ISR). In the present study, we evaluated the involvement of the innate immune response in the K165-mediated protection of Arabidopsis against V. dahliae. Tests with Arabidopsis mutants impaired in several regulators of the early steps of the innate immune responses, including fls2, efr-1, bak1-4, mpk3, mpk6, wrky22, and wrky29 showed that FLS2 and WRKY22 have a central role in the K165-triggered ISR, while EFR1, MPK3, and MPK6 are possible susceptibility factors for V. dahliae and bak1 shows a tolerance phenomenon. The resistance induced by strain K165 is dependent on both salicylate and jasmonate-dependent defense pathways, as evidenced by an increased transient accumulation of PR1 and PDF1.2 transcripts in the aerial parts of infected plants treated with strain K165.

  14. PENGARUH INFESTASI VERTICILLIUM LECANII TERHADAP KEPARAHAN PENYAKIT KARAT DAUN KOPI PADA TANAMAN DAN KETERJADIAN KOLONINYA PADA DAUN

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    Cipta Ginting .

    2011-11-01

    Full Text Available Effect of infestation of Verticillium lecanii on the severity of coffee leaf rust on plants and the incidence of its colony on coffee leaves.  V. lecanii often antagonizes H. vastatrix on lesions caused by the pathogen on coffee leaves in the field.  The objectives of this study were to evaluate the efficacy of V. lecanii to control disease severity on coffee plants in a green house and to compare the incidence of V. lecanii colonies on coffee leaves naturally infested by V. lecanii in the field and that in the leaves with additional infestation in the laboratory.  For efficacy test and additional infestation in the laboratory, conidia were harvested from YMPD medium.  Suspension with 107 conidia ml-1 was sprayed into coffee foliage until both surfaces of the leaves were saturated with the suspension.  After the leaves were air dried, they were sprayed with urediospores (104 ml-1.  The results show that disease severity in plants treated with V. lecanii and control plants was not significantly different (P < 0.05, except that obtained at the fourth week when disease severity on the treated plants was significantly lower that that in control plants.  The number of V. lecanii colonies on coffee leaves naturally infested with the biocontrol agent was not increased by additional infestation with V. lecanii conidia in the laboratory.

  15. Discovery and identification of candidate genes from the chitinase gene family for Verticillium dahliae resistance in cotton.

    Science.gov (United States)

    Xu, Jun; Xu, Xiaoyang; Tian, Liangliang; Wang, Guilin; Zhang, Xueying; Wang, Xinyu; Guo, Wangzhen

    2016-06-29

    Verticillium dahliae, a destructive and soil-borne fungal pathogen, causes massive losses in cotton yields. However, the resistance mechanism to V. dahilae in cotton is still poorly understood. Accumulating evidence indicates that chitinases are crucial hydrolytic enzymes, which attack fungal pathogens by catalyzing the fungal cell wall degradation. As a large gene family, to date, the chitinase genes (Chis) have not been systematically analyzed and effectively utilized in cotton. Here, we identified 47, 49, 92, and 116 Chis from four sequenced cotton species, diploid Gossypium raimondii (D5), G. arboreum (A2), tetraploid G. hirsutum acc. TM-1 (AD1), and G. barbadense acc. 3-79 (AD2), respectively. The orthologous genes were not one-to-one correspondence in the diploid and tetraploid cotton species, implying changes in the number of Chis in different cotton species during the evolution of Gossypium. Phylogenetic classification indicated that these Chis could be classified into six groups, with distinguishable structural characteristics. The expression patterns of Chis indicated their various expressions in different organs and tissues, and in the V. dahliae response. Silencing of Chi23, Chi32, or Chi47 in cotton significantly impaired the resistance to V. dahliae, suggesting these genes might act as positive regulators in disease resistance to V. dahliae.

  16. Perturbations in the Primary Metabolism of Tomato and Arabidopsis thaliana Plants Infected with the Soil-Borne Fungus Verticillium dahliae.

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    Anja Buhtz

    Full Text Available The hemibiotrophic soil-borne fungus Verticillium dahliae is a major pathogen of a number of economically important crop species. Here, the metabolic response of both tomato and Arabidopsis thaliana to V. dahliae infection was analysed by first using non-targeted GC-MS profiling. The leaf content of both major cell wall components glucuronic acid and xylose was reduced in the presence of the pathogen in tomato but enhanced in A. thaliana. The leaf content of the two tricarboxylic acid cycle intermediates fumaric acid and succinic acid was increased in the leaf of both species, reflecting a likely higher demand for reducing equivalents required for defence responses. A prominent group of affected compounds was amino acids and based on the targeted analysis in the root, it was shown that the level of 12 and four free amino acids was enhanced by the infection in, respectively, tomato and A. thaliana, with leucine and histidine being represented in both host species. The leaf content of six free amino acids was reduced in the leaf tissue of diseased A. thaliana plants, while that of two free amino acids was raised in the tomato plants. This study emphasizes the role of primary plant metabolites in adaptive responses when the fungus has colonized the plant.

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

    Institute of Scientific and Technical Information of China (English)

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

    2016-01-01

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

  18. Genotyping-by-sequencing-based genome-wide association studies on Verticillium wilt resistance in autotetraploid alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Yu, Long-Xi; Zheng, Ping; Zhang, Tiejun; Rodringuez, Jonas; Main, Dorrie

    2017-02-01

    Verticillium wilt (VW) is a fungal disease that causes severe yield losses in alfalfa. The most effective method to control the disease is through the development and use of resistant varieties. The identification of marker loci linked to VW resistance can facilitate breeding for disease-resistant alfalfa. In the present investigation, we applied an integrated framework of genome-wide association with genotyping-by-sequencing (GBS) to identify VW resistance loci in a panel of elite alfalfa breeding lines. Phenotyping was performed by manual inoculation of the pathogen to healthy seedlings, and scoring for disease resistance was carried out according to the standard test of the North America Alfalfa Improvement Conference (NAAIC). Marker-trait association by linkage disequilibrium identified 10 single nucleotide polymorphism (SNP) markers significantly associated with VW resistance. Alignment of the SNP marker sequences to the M. truncatula genome revealed multiple quantitative trait loci (QTLs). Three, two, one and five markers were located on chromosomes 5, 6, 7 and 8, respectively. Resistance loci found on chromosomes 7 and 8 in the present study co-localized with the QTLs reported previously. A pairwise alignment (blastn) using the flanking sequences of the resistance loci against the M. truncatula genome identified potential candidate genes with putative disease resistance function. With further investigation, these markers may be implemented into breeding programmes using marker-assisted selection, ultimately leading to improved VW resistance in alfalfa. PUBLISHED 2016. THIS ARTICLE IS A U.S. GOVERNMENT WORK AND IS IN THE PUBLIC DOMAIN IN THE USA.

  19. Functional analysis of autophagy genes via Agrobacterium-mediated transformation in the vascular Wilt fungus Verticillium dahliae.

    Science.gov (United States)

    Zhou, Lei; Zhao, Jun; Guo, Wangzhen; Zhang, Tianzhen

    2013-08-20

    Autophagy is a widely conserved intracellular process for degradation and recycling of proteins, organelles and cytoplasm in eukaryotic organisms and is now emerging as an important process in foliar infection by many plant pathogenic fungi. However, the role of autophagy in soil-borne fungal physiology and infection biology is poorly understood. Here, we report the establishment of an Agrobacterium tumefaciens-mediated transformation (ATMT) system and its application to investigate two autophagy genes, VdATG8 and VdATG12, by means of targeted gene replacement and complementation. Transformation of a cotton-infecting Verticillium dahliae strain Vd8 with a novel binary vector pCOM led to the production of 384 geneticin-resistant transformants per 1 × 10(6) conidia. V. dahliae mutants lacking either VdATG8 or VdATG12 exhibited reduced conidiation and impaired aerial hyphae production. Disease development on Arabidopsis plants was slightly delayed when inoculated with VdATG8 or VdATG12 gene deletion mutants, compared with the wild-type and gene complemented strains. Surprisingly, in vitro inoculation with unimpaired roots revealed that the abilities of root invasion were not affected in gene deletion mutants. These results indicate that autophagy is necessary for aerial hyphae development and plant colonization but not for root infection in V. dahliae.

  20. In vitro effect of essential oils from aromatic and medicinal plants on mushroom pathogens: Verticillium fungicola var. fungicola, Mycogone perniciosa, and Cladobotryum sp.

    Directory of Open Access Journals (Sweden)

    Tanović Brankica

    2009-01-01

    Full Text Available Lavender, anise, chamomile, fennel, geranium, oregano, parsley, and sage essential oils were tested for their effectiveness against mushroom pathogens: Verticillium fungicola var. fungicola, Mycogone perniciosa, and Cladobotryum sp. Isolates were exposed to the volatile phase of the oils and then ventilated in order to determine if the effect of the oil was lethal to the pathogen. Oregano and geranium oils were the most toxic, having a fungicidal effect at 0.02-0.08 μl/ml of air, depending on the pathogen. Oregano oil was characterized by high content of carvacrol and thymol, while citranelol and geraniol were the main components of geranium oil.

  1. CEREALS ASSESSMENT TOWARDS CONTAMINATION OF PHYTOPATHOGENIC FUNGI IN FOREST-STEPPE AREA OF UKRAINE

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    V. B. Yekimova

    2014-11-01

    Full Text Available The usage of high-quality seed, not affected by pathogens and fungi promote the high and stable yields. The condition of seeds determines their germination, seedling damage and adult plants, which ultimately affects the yield and on their quality. The significant risk of seed infection was registered in years with high humidity in the pre-harvest and harvest period. Therefore, along with the traditional seed control it is necessary to conduct phytopahtology expertise, allowing to identify the species composition of the microflora of seeds and the degree of infection with different pathogens. Smut disease - a solid and loose smut of wheat, hard and loose smut of barley - cause crop losses are clear - in the form of the destruction of the ear, and hidden - in the form of a reduction in seed germination, reduce winter hardiness, plant growth inhibition. Pathogens root rot causing blight, Fusarium and Helminthosporium that insignificantly demand on environmental conditions and extremely plastic. They have large set of enzymes that can exist on a variety of substrates, and therefore are widely distributed in nature and cause considerable damage to crops. Especially significant losses occur if the humidity during the ripening grain observed for several seasons, which leads to accumulation of the infection naturally. The aim of research was to assess the contamination of grain of spring wheat and barley phytopathogenic fungi in one of the main grain regions of Ukraine - the southern forest. The research conducted during the summer 2014 and compared with the results of previous years. Samples were taken from different plots. Seed contamination of samples by various phytopathogenic fungi was determined by the number of infected kernels per 100 seed sample. Frequency of registered species was recorded. For every studied sample we set the percentage of species. Analysis of the grain on the fungal infection and avdelenie in pure culture was performed

  2. CEREALS ASSESSMENT TOWARDS CONTAMINATION OF PHYTOPATHOGENIC FUNGI IN FOREST-STEPPE AREA OF UKRAINE

    Directory of Open Access Journals (Sweden)

    Yekimova V. B.

    2014-12-01

    Full Text Available The usage of high-quality seed, not affected by pathogens and fungi promote the high and stable yields. The condition of seeds determines their germination, seedling damage and adult plants, which ultimately affects the yield and on their quality. The significant risk of seed infection was registered in years with high humidity in the pre-harvest and harvest period. Therefore, along with the traditional seed control it is necessary to conduct phytopahtology expertise, allowing to identify the species composition of the microflora of seeds and the degree of infection with different pathogens. Smut disease - a solid and loose smut of wheat, hard and loose smut of barley - cause crop losses are clear - in the form of the destruction of the ear, and hidden - in the form of a reduction in seed germination, reduce winter hardiness, plant growth inhibition. Pathogens root rot causing blight, Fusarium and Helminthosporium that insignificantly demand on environmental conditions and extremely plastic. They have large set of enzymes that can exist on a variety of substrates, and therefore are widely distributed in nature and cause considerable damage to crops. Especially significant losses occur if the humidity during the ripening grain observed for several seasons, which leads to accumulation of the infection naturally. The aim of research was to assess the contamination of grain of spring wheat and barley phytopathogenic fungi in one of the main grain regions of Ukraine - the southern forest. The research conducted during the summer 2014 and compared with the results of previous years. Samples were taken from different plots. Seed contamination of samples by various phytopathogenic fungi was determined by the number of infected kernels per 100 seed sample. Frequency of registered species was recorded. For every studied sample we set the percentage of species. Analysis of the grain on the fungal infection and avdelenie in pure culture was performed

  3. Impact of phytopathogen infection and extreme weather stress on internalization of Salmonella Typhimurium in lettuce.

    Science.gov (United States)

    Ge, Chongtao; Lee, Cheonghoon; Nangle, Ed; Li, Jianrong; Gardner, David; Kleinhenz, Matthew; Lee, Jiyoung

    2014-01-03

    Internalization of human pathogens, common in many types of fresh produce, is a threat to human health since the internalized pathogens cannot be fully inactivated/removed by washing with water or sanitizers. Given that pathogen internalization can be affected by many environmental factors, this study was conducted to investigate the influence of two types of plant stress on the internalization of Salmonella Typhimurium in iceberg lettuce during pre-harvest. The stresses were: abiotic (water stress induced by extreme weather events) and biotic (phytopathogen infection by lettuce mosaic virus [LMV]). Lettuce with and without LMV infection were purposefully contaminated with green fluorescence protein-labeled S. Typhimurium on the leaf surfaces. Lettuce was also subjected to water stress conditions (drought and storm) which were simulated by irrigating with different amounts of water. The internalized S. Typhimurium in the different parts of the lettuce were quantified by plate count and real-time quantitative PCR and confirmed with a laser scanning confocal microscope. Salmonella internalization occurred under the conditions outlined above; however internalization levels were not significantly affected by water stress alone. In contrast, the extent of culturable S. Typhimurium internalized in the leafy part of the lettuce decreased when infected with LMV under water stress conditions and contaminated with high levels of S. Typhimurium. On the other hand, LMV-infected lettuce showed a significant increase in the levels of culturable bacteria in the roots. In conclusion, internalization was observed under all experimental conditions when the lettuce surface was contaminated with S. Typhimurium. However, the extent of internalization was only affected by water stress when lettuce was infected with LMV.

  4. Leaf-cutting ant fungi produce cell wall degrading pectinase complexes reminiscent of phytopathogenic fungi

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    Boomsma Jacobus J

    2010-12-01

    those normally found in phytopathogens.

  5. Septins from the phytopathogenic fungus Ustilago maydis are required for proper morphogenesis but dispensable for virulence.

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    Isabel Alvarez-Tabarés

    Full Text Available BACKGROUND: Septins are a highly conserved family of GTP-binding proteins involved in multiple cellular functions, including cell division and morphogenesis. Studies of septins in fungal cells underpin a clear correlation between septin-based structures and fungal morphology, providing clues to understand the molecular frame behind the varied morphologies found in fungal world. METHODOLOGY/PRINCIPAL FINDINGS: Ustilago maydis genome has the ability to encode four septins. Here, using loss-of-function as well as GFP-tagged alleles of these septin genes, we investigated the roles of septins in the morphogenesis of this basidiomycete fungus. We described that septins in U. maydis could assemble into at least three different structures coexisting in the same cell: bud neck collars, band-like structures at the growing tip, and long septin fibers that run from pole to pole near the cell cortex. We also found that in the absence of septins, U. maydis cells lost their elongated shape, became wider at the central region and ended up losing their polarity, pointing to an important role of septins in the morphogenesis of this fungus. These morphological defects were alleviated in the presence of an osmotic stabilizer suggesting that absence of septins affected the proper formation of the cell wall, which was coherent with a higher sensitivity of septin defective cells to drugs that affect cell wall construction as well as exocytosis. As U. maydis is a phytopathogen, we analyzed the role of septins in virulence and found that in spite of the described morphological defects, septin mutants were virulent in corn plants. CONCLUSIONS/SIGNIFICANCE: Our results indicated a major role of septins in morphogenesis in U. maydis. However, in contrast to studies in other fungal pathogens, in which septins were reported to be necessary during the infection process, we found a minor role of septins during corn infection by U. maydis.

  6. Ag doped hollow TiO2 nanoparticles as an effective green fungicide against Fusarium solani and Venturia inaequalis phytopathogens

    Science.gov (United States)

    Sankar Boxi, Siddhartha; Mukherjee, Khushi; Paria, Santanu

    2016-02-01

    Chemical-based pesticides are widely used in agriculture to protect crops from insect infestation and diseases. However, the excessive use of highly toxic pesticides causes several human health (neurological, tumor, cancer) and environmental problems. Therefore nanoparticle-based green pesticides have become of special importance in recent years. The antifungal activities of pure and Ag doped (solid and hollow) TiO2 nanoparticles are studied against two potent phytopathogens, Fusarium solani (which causes Fusarium wilt disease in potato, tomato, etc) and Venturia inaequalis (which causes apple scab disease) and it is found that hollow nanoparticles are more effective than the other two. The antifungal activities of the nanoparticles were further enhanced against these two phytopathogens under visible light exposure. The fungicidal effect of the nanoparticles depends on different parameters, such as particle concentration and the intensity of visible light. The minimum inhibitory dose of the nanoparticles for V. inaequalis and F. solani are 0.75 and 0.43 mg/plate. The presence of Ag as a dopant helps in the formation of stable Ag-S and disulfide bonds (R-S-S-R) in cellular protein, which leads to cell damage. During photocatalysis generated •OH radicals loosen the cell wall structure and this finally leads to cell death. The mechanisms of the fungicidal effect of nanoparticles against these two phytopathogens are supported by biuret and triphenyl tetrazolium chloride analyses and field emission electron microscopy. Apart from the fungicidal effect, at a very low dose (0.015 mg/plate) the nanoparticles are successful in arresting production of toxic napthoquinone pigment for F. solani which is related to the fungal pathogenecity. The nanoparticles are found to be effective in protecting potatoes affected by F. solani or other fungi from spoiling.

  7. Ag doped hollow TiO2 nanoparticles as an effective green fungicide against Fusarium solani and Venturia inaequalis phytopathogens.

    Science.gov (United States)

    Boxi, Siddhartha Sankar; Mukherjee, Khushi; Paria, Santanu

    2016-02-26

    Chemical-based pesticides are widely used in agriculture to protect crops from insect infestation and diseases. However, the excessive use of highly toxic pesticides causes several human health (neurological, tumor, cancer) and environmental problems. Therefore nanoparticle-based green pesticides have become of special importance in recent years. The antifungal activities of pure and Ag doped (solid and hollow) TiO2 nanoparticles are studied against two potent phytopathogens, Fusarium solani (which causes Fusarium wilt disease in potato, tomato, etc) and Venturia inaequalis (which causes apple scab disease) and it is found that hollow nanoparticles are more effective than the other two. The antifungal activities of the nanoparticles were further enhanced against these two phytopathogens under visible light exposure. The fungicidal effect of the nanoparticles depends on different parameters, such as particle concentration and the intensity of visible light. The minimum inhibitory dose of the nanoparticles for V. inaequalis and F. solani are 0.75 and 0.43 mg/plate. The presence of Ag as a dopant helps in the formation of stable Ag-S and disulfide bonds (R-S-S-R) in cellular protein, which leads to cell damage. During photocatalysis generated (•)OH radicals loosen the cell wall structure and this finally leads to cell death. The mechanisms of the fungicidal effect of nanoparticles against these two phytopathogens are supported by biuret and triphenyl tetrazolium chloride analyses and field emission electron microscopy. Apart from the fungicidal effect, at a very low dose (0.015 mg/plate) the nanoparticles are successful in arresting production of toxic napthoquinone pigment for F. solani which is related to the fungal pathogenecity. The nanoparticles are found to be effective in protecting potatoes affected by F. solani or other fungi from spoiling.

  8. Screening and Identification of Cotton Verticillium Wilt Antagonistic Bacteria Strain 7-30%棉花黄萎病拮抗细菌7-30菌株的筛选与鉴定

    Institute of Scientific and Technical Information of China (English)

    雷白时; 姜军坡; 王伟; 张冬冬; 王全; 朱宝成

    2008-01-01

    [Objective]The aim of this study was to screen the antagonistic spore-forming bacteria of Verticillium dahliae and identify its physiological and biochemical characteristics. [Method]Taking the cotton verticillium wilt pathogen Verticillium dahliae V-190 as the test microorganism, the antagonistic spore-forming bacteria were screened. Through the preliminary screening and secondary screening, an antagonistic bacteria strain 7-30 with stronger antibacterial activity was obtained, and its morphological characteristics, physiological and biochemical characteristics were also identified. [Result]84 antagonistic bacteria strains were isolated from soil in various places by the preliminary screening. Especially, 18 strains with better antagonistic ability were screened again, so an antagonistic bacteria strain 7-30 with the diameter of inhibition zone 18.9 mm and stronger antibacterial activity was obtained. According to its morphological characteristics, physiological and biochemical characteristics, the antagonistic bacteria strain 7-30 was identified as Bacillus subtilis primarily. [Conclusion]The strain 7-30 was obtained as the antagonistic spore-forming bacteria of Verticillium dahliae.

  9. Induced Resistance in Tomato Plants against Verticillium Wilt by the Binuclear Nickel Coordination Complex of the Ligand 2,5-Bis(pyridin-2-yl)-1,3,4-thiadiazole.

    Science.gov (United States)

    Zine, Hanane; Rifai, Lalla Aicha; Faize, Mohamed; Bentiss, Fouad; Guesmi, Salaheddine; Laachir, Abdelhakim; Smaili, Amal; Makroum, Kacem; Sahibed-Dine, Abdelaziz; Koussa, Tayeb

    2016-04-06

    Verticillium wilt caused by Verticillium dahliae is a major limiting factor for tomato production. The objective of this study was to evaluate the effectiveness of ligand 2,5-bis(pyridin-2-yl)-1,3,4-thiadiazole (L) and its complex bis[μ-2,5-bis(pyridin-2-yl)-1,3,4-thiadiazole-κ(4)N(2),N(3):N(4),N(5)]bis[dihydrato-κO)nickel(II)] as activators of plant defenses in controlling Verticillium wilt. In the greenhouse, they protected tomato plants against V. dahliae when they were applied twice as foliar sprays at 100 μg mL(-1). A synergistic effect was observed between the ligand L and the transition metal Ni, with disease incidence reduced by 38% with L and 57% with Ni2L2. Verticillium wilt foliar symptoms and vascular browning index were reduced by 82% for L and 95% for Ni2L2. This protection ability was associated with the induction of an oxidative burst and the activation of the total phenolic content as well as potentiation of the activity of peroxidase and polyphenol oxidase. These results demonstrated that L and Ni2L2 can be considered as new activators of plant defense responses.

  10. Crescimento e esporulação de isolados de Verticillium lecanii sob diferentes condições nutricionais

    Directory of Open Access Journals (Sweden)

    Barbosa Claudio Camargo

    2002-01-01

    Full Text Available O objetivo deste trabalho foi determinar as melhores condições de crescimento e esporulação de dois isolados de Verticillium lecanii (Zimm. Viégas, cultivados em vários meios de cultura, fontes de C, fontes orgânicas e inorgânicas de N e relações C:N. Após 20 dias de cultivo, BDA e Meio Completo mostraram ser os melhores meios de cultura, e a lactose e o amido foram as fontes de C que mais favoreceram o crescimento dos isolados JAB 02 e JAB 45, respectivamente; baixa esporulação foi apresentada por JAB 02, enquanto JAB 45 obteve boa esporulação em glicose e maltose. JAB 02 desenvolveu melhor quando a fonte orgânica de N era casitona; JAB 45 obteve bom desenvolvimento em presença de casitona e peptona, mas houve pouca esporulação em presença de triptona e caseína hidrolisada. As fontes inorgânicas de N que favoreceram o melhor desenvolvimento dos isolados foram (NH42HPO4 e NaNO3, porém, JAB 02 apresentou baixa esporulação. Entre as relações C:N analisadas, JAB 02 obteve melhor crescimento na relação 60:1 e JAB 45 nas relações 5:1, 10:1 e 20:1; baixa esporulação foi obtida por JAB 02 nas relações testadas, mas não se verificou efeito da relação C:N do meio sobre a esporulação dos isolados.

  11. Identification of MiRNA from eggplant (Solanum melongena L. by small RNA deep sequencing and their response to Verticillium dahliae infection.

    Directory of Open Access Journals (Sweden)

    Liu Yang

    Full Text Available MiRNAs are a class of non-coding small RNAs that play important roles in the regulation of gene expression. Although plant miRNAs have been extensively studied in model systems, less is known in other plants with limited genome sequence data, including eggplant (Solanum melongena L.. To identify miRNAs in eggplant and their response to Verticillium dahliae infection, a fungal pathogen for which clear understanding of infection mechanisms and effective cure methods are currently lacking, we deep-sequenced two small RNA (sRNA libraries prepared from mock-infected and infected seedlings of eggplants. Specifically, 30,830,792 reads produced 7,716,328 unique miRNAs representing 99 known miRNA families that have been identified in other plant species. Two novel putative miRNAs were predicted with eggplant ESTs. The potential targets of the identified known and novel miRNAs were also predicted based on sequence homology search. It was observed that the length distribution of obtained sRNAs and the expression of 6 miRNA families were obviously different between the two libraries. These results provide a framework for further analysis of miRNAs and their role in regulating plant response to fungal infection and Verticillium wilt in particular.

  12. Colonization process of olive tissues by Verticillium dahliae and its in planta interaction with the biocontrol root endophyte Pseudomonas fluorescens PICF7

    Science.gov (United States)

    Prieto, Pilar; Navarro‐Raya, Carmen; Valverde‐Corredor, Antonio; Amyotte, Stefan G.; Dobinson, Katherine F.; Mercado‐Blanco, Jesús

    2009-01-01

    Summary The colonization process of Olea europaea by the defoliating pathotype of Verticillium dahliae, and the in planta interaction with the endophytic, biocontrol strain Pseudomonas fluorescens PICF7 were determined. Differential fluorescent protein tagging was used for the simultaneous visualization of P. fluorescens PICF7 and V. dahliae in olive tissues. Olive plants were bacterized with PICF7 and then transferred to V. dahliae‐infested soil. Monitoring olive colonization events by V. dahliae and its interaction with PICF7 was conducted using a non‐gnotobiotic system, confocal laser scanner microscopy and tissue vibratoming sections. A yellow fluorescently tagged V. dahliae derivative (VDAT‐36I) was obtained by Agrobacterium tumefaciens‐mediated transformation. Isolate VDAT‐36I quickly colonized olive root surface, successfully invaded root cortex and vascular tissues via macro‐ and micro‐breakages, and progressed to the aerial parts of the plant through xylem vessel cells. Strain PICF7 used root hairs as preferred penetration site, and once established on/in root tissues, hindered pathogen colonization. For the first time using this approach, the entire colonization process of a woody plant by V. dahliae is reported. Early and localized root surface and root endophytic colonization by P. fluorescens PICF7 is needed to impair full progress of verticillium wilt epidemics in olive. PMID:21255281

  13. Ectopic expression of a Ve homolog VvVe gene from Vitis vinifera enhances defense response to Verticillium dahliae infection in tobacco.

    Science.gov (United States)

    Tang, Juan; Lin, Jing; Yang, Yuwen; Chen, Tianzi; Ling, Xitie; Zhang, Baolong; Chang, Youhong

    2016-01-15

    Verticillium wilt is a soil borne disease that can cause devastating losses to the production of many economically important crops. A Ve1 homologous gene responding to Verticillium dahliae infection was identified in Vitis vinifera cv. "HeiFeng" by semi-quantitative reverse transcription polymerase chain reaction and was designated as VvVe. The overexpression of VvVe in transgenic Nicotiana benthamiana plants significantly enhanced the resistance to isolate V991 of V. dahliae when compared with the wild type plants. The expressions of defense-related genes including the salicylic acid regulated gene pathogen-related 1 (PR1) but not PR2, the ethylene- and jasmonic acid-regulated genes ethylene response factor 1 (ERF1) and lipoxygenase (LOX) were significantly increased due to over expression of VvVe. And greater accumulation of active oxygen, callose and phenylalanine-ammonia lyase were observed in the leaves of transgenic VvVe tobacco plants than the wild type when under infection by V. dahliae. Moreover, the hypersensitive response mimicking cell death was exclusively occurred in the transgenic VvVe tobacco plants but not in the wild type. Taken together, the VvVe gene is a Ve1 like gene which involves in the signal cascade of salicylic acid, jasmonate, and ethylene defense pathways and enhances defense response to V. dahliae infection in the transgenic tobacco. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. 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).

    Science.gov (United States)

    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 biocontrol agent against V. dahliae and growth elicitor in tomato.

  15. Inhibitory effect of Thymus vulgaris and Origanum vulgare essential oils on virulence factors of phytopathogenic Pseudomonas syringae strains.

    Science.gov (United States)

    Carezzano, M E; Sotelo, J P; Primo, E; Reinoso, E B; Paletti Rovey, M F; Demo, M S; Giordano, W F; Oliva, M de Las M

    2017-07-01

    Pseudomonas syringae is a phytopathogenic bacterium that causes lesions in leaves during the colonisation process. The damage is associated with production of many virulence factors, such as biofilm and phytotoxins. The essential oils of Thymus vulgaris (thyme) and Origanum vulgare (oregano) have been demonstrated to inhibit P. syringae. The aim of this study was to investigate the effects of T. vulgaris and O. vulgare essential oils on production of virulence factors of phytopathogenic P. syringae strains, including anti-biofilm and anti-toxins activities. The broth microdilution method was used for determination of MIC and biofilm inhibition assays. Coronatine, syringomycin and tabtoxin were pheno- and genotypically evaluated. Both oils showed good inhibitory activity against P. syringae, with MIC values from 1.43 to 11.5 mg·ml(-1) for thyme and 5.8 to 11.6 mg·ml(-1) for oregano. Biofilm formation, production of coronatine, syringomycin and tabtoxin were inhibited by thyme and oregano essential oil in most strains. The results presented here are promising, demonstrating the bactericidal activity and reduction of virulence factor production after treatment with thyme and oregano oil, providing insight into how they exert their antibacterial activity. These natural products could be considered in the future for the control of diseases caused by P. syringae. © 2017 German Botanical Society and The Royal Botanical Society of the Netherlands.

  16. Selection of endophytic fungi from comfrey (Symphytum officinale L.) for in vitro biological control of the phytopathogen Sclerotinia sclerotiorum (Lib.).

    Science.gov (United States)

    Rocha, Rafaeli; da Luz, Daniela Eleutério; Engels, Cibelle; Pileggi, Sônia Alvim Veiga; de Souza Jaccoud Filho, David; Matiello, Rodrigo Rodrigues; Pileggi, Marcos

    2009-01-01

    Biological control consists of using one organism to attack another that may cause economic damage to crops. Integrated Pest Management (IPM) is a very common strategy. The white mold produced by Sclerotinia sclerotiorum (Lib.) causes considerable damage to bean crops. This fungus is a soil inhabitant, the symptoms of which are characterized by water-soaked lesions covered by a white cottony fungal growth on the soil surface and/or the host plant. Possible biological control agents taken from plants are being investigated as phytopathogen inhibitors. These are endophytic microorganisms that inhabit the intercellular spaces of vegetal tissues and are often responsible for antimicrobial production. The objective of the present study was to select endophytic fungi isolated from comfrey (Symphytum officinale L.) leaves with in vitro antagonist potential against the phytopathogenic fungus S. sclerotiorum. Twelve isolates of endophytic fungi and a pathogenic strain of S. sclerotiorum were used in the challenge method. With the aid of this method, four endophytes with the best antagonistic activity against S. sclerotiorum were selected. Pathogen growth inhibition zones were considered indicative of antibiosis. The percentages of pathogenic mycelia growth were measured both with and without the antagonist, resulting in growth reductions of 46.7% to 50.0% for S. sclerotiorum. These analyses were performed by evaluating the endophytic/pathogenic mycelia growth in mm/day over an eight-day period of antagonistic tests.

  17. Fasciation induction by the phytopathogen Rhodococcus fascians depends upon a linear plasmid encoding a cytokinin synthase gene.

    Science.gov (United States)

    Crespi, M; Messens, E; Caplan, A B; van Montagu, M; Desomer, J

    1992-01-01

    Rhodococcus fascians is a nocardiform bacteria that induces leafy galls (fasciation) on dicotyledonous and several monocotyledonous plants. The wild-type strain D188 contained a conjugative, 200 kb linear extrachromosomal element, pFiD188. Linear plasmid-cured strains were avirulent and reintroduction of this linear element restored virulence. Pulsed field electrophoresis indicated that the chromosome might also be a linear molecule of 4 megabases. Three loci involved in phytopathogenicity have been identified by insertion mutagenesis of this Fi plasmid. Inactivation of the fas locus resulted in avirulent strains, whereas insertions in the two other loci affected the degree of virulence, yielding attenuated (att) and hypervirulent (hyp) bacteria. One of the genes within the fas locus encoded an isopentenyltranferase (IPT) with low homology to analogous proteins from Gram-negative phytopathogenic bacteria. IPT activity was detected after expression of this protein in Escherichia coli cells. In R.fascians, ipt expression could only be detected in bacteria induced with extracts from fasciated tissue. R.fascians strains without the linear plasmid but containing this fas locus alone could not provoke any phenotype on plants, indicating additional genes from the linear plasmid were also essential for virulence. These studies, the first genetic analysis of the interaction of a Gram-positive bacterium with plants, suggest that a novel mechanism for plant tumour induction has evolved in R.fascians independently from the other branches of the eubacteria. Images PMID:1547783

  18. Synthesis of pyrazolo[1,5-a]pyrimidine derivatives and their antifungal activities against phytopathogenic fungi in vitro.

    Science.gov (United States)

    Zhang, Jin; Peng, Ju-Fang; Bai, Yu-Bin; Wang, Ping; Wang, Tao; Gao, Jin-Ming; Zhang, Zun-Ting

    2016-11-01

    5,6-Diarylpyrazolo[1,5-a]pyrimidines (3) and 6,7-diarylpyrazolo[1,5-a]pyrimidines (4) were chemoselectively synthesized by the condensation of isoflavone (1) and 3-aminopyrazole (2). 5,6-Diarylpyrazolo[1,5-a]pyrimidines (3) were obtained via microwave irradiation, and 6,7-diarylpyrazolo[1,5-a]pyrimidines (4) were obtained via conventional heating. In addition, the pyrimidine derivatives 3 and 4 were evaluated against five phytopathogenic fungi (Cytospora sp., Colletotrichum gloeosporioides, Botrytis cinerea, Alternaria solani, and Fusarium solani) using the mycelium growth rate method. Some of them were effective in inhibiting the growth of the five phytopathogenic fungi. For instance, 6,7-diarylpyrazolo[1,5-a]pyrimidines (4j) inhibited the growth of A. solani with an [Formula: see text] value of 17.11 [Formula: see text], and 6,7-diarylpyrazolo[1,5-a]pyrimidines (4h) inhibited the growth of both Cytospora sp. and F. solani with [Formula: see text] values of 27.32 and 21.04 [Formula: see text], respectively. A chemoselective synthesis of 5,6-pyrazolo[1,5-a]pyrimidines 3 derivatives in excellent yields was performed under microwave irradiation and 6,7-pyrazolo[1,5-a]pyrimidines 4 were also prepared using heating method. The antifungal properties of 3 and 4 were tested against Cytospora sp., Colletotrichum gloeosporioides, Botrytis cinerea, Alternaria solani, and Fusarium solani.

  19. Identification and characterization of integron-mediated antibiotic resistance in the phytopathogen Xanthomonas oryzae pv. oryzae.

    Directory of Open Access Journals (Sweden)

    Ying Xu

    report of resistance integron in a phytopathogenic bacteria.

  20. Pathogen and biological contamination management in plant tissue culture: phytopathogens, vitro pathogens, and vitro pests.

    Science.gov (United States)

    Cassells, Alan C

    2012-01-01

    The ability to establish and grow plant cell, organ, and tissue cultures has been widely exploited for basic and applied research, and for the commercial production of plants (micro-propagation). Regardless of whether the application is for research or commerce, it is essential that the cultures be established in vitro free of biological contamination and be maintained as aseptic cultures during manipulation, growth, and storage. The risks from microbial contamination are spurious experimental results due to the effects of latent contaminants or losses of valuable experimental or commercial cultures. Much of the emphasis in culture contamination management historically focussed on the elimination of phytopathogens and the maintenance of cultures free from laboratory contamination by environmental bacteria, fungi (collectively referred to as "vitro pathogens", i.e. pathogens or environmental micro-organisms which cause culture losses), and micro-arthropods ("vitro pests"). Microbial contamination of plant tissue cultures is due to the high nutrient availability in the almost universally used Murashige and Skoog (Physiol Plant 15:473-497, 1962) basal medium or variants of it. In recent years, it has been shown that many plants, especially perennials, are at least locally endophytically colonized intercellularly by bacteria. The latter, and intracellular pathogenic bacteria and viruses/viroids, may pass latently into culture and be spread horizontally and vertically in cultures. Growth of some potentially cultivable endophytes may be suppressed by the high salt and sugar content of the Murashige and Skoog basal medium and suboptimal temperatures for their growth in plant tissue growth rooms. The management of contamination in tissue culture involves three stages: disease screening (syn. disease indexing) of the stock plants with disease and endophyte elimination where detected; establishment and pathogen and contaminant screening of established initial cultures

  1. Antifungal activity of essential oil isolated from Ocimum gratissimum L. (eugenol chemotype against phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    Terezinha de Jesus Faria

    2006-11-01

    Full Text Available An investigation of antifungal activity of the essential oil obtained by steam-distillation (1.1% w/w of the aerial parts of Ocimum gratissimum and of an ethanolic extract from the steam-distillation residue was carried out using the agar diffusion method. The results revealed that the essential oil inhibited the growth of all fungi tested, including the phytopathogens, Botryosphaeria rhodina, Rhizoctonia sp. and two strains of Alternaria sp., while the extract from the residue was inactive. The essential oil was subjected to TLC bioautography used to detect fungitoxic constituents. The compound that showed antifungal activity was isolated and identified as eugenol. GC/MS analysis showed that eugenol was the main constituent of the essential oil studied. The antifungal activity of eugenol was evaluated against a species of Alternaria isolated from tomato (A1 and Penicillium chrysogenum. The minimal inhibitory concentrations of eugenol were 0.16 and 0.31 mg/disc for Alternaria sp. (A1 and P. chrysogenum, respectively.O óleo essencial resultante da destilação por arraste a vapor das partes aéreas de Ocimum gratissimum e o extrato etanólico obtido do resíduo da destilação foram avaliados quanto à atividade antifúngica, utilizando-se o método de difusão em agar. O óleo essencial inibiu o crescimento de todos os fungos testados, incluindo os fitopatogênicos Botryosphaeria rhodina e duas espécies de Alternaria sp, enquanto que o extrato do resíduo da destilação não apresentou atividade. O óleo essencial foi, então, submetido ao método de bioautografia em TLC para detecção do composto ativo. O componente ativo foi isolado e identificado através da análise por cromatografia gasosa acoplada à espectrometria de massas como o eugenol, constituinte majoritário do óleo estudado. Ensaios de atividade antifúngica revelaram a atividade do eugenol contra Alternaria isolada de tomate (A1 e Penicillium chrysogenum. As concentra

  2. Sequential Delivery of Host-Induced Virulence Effectors by Appressoria and Intracellular Hyphae of the Phytopathogen Colletotrichum higginsianum

    Science.gov (United States)

    Kleemann, Jochen; Neumann, Ulla; van Themaat, Emiel Ver Loren; van der Does, H. Charlotte; Hacquard, Stéphane; Stüber, Kurt; Will, Isa; Schmalenbach, Wolfgang; Schmelzer, Elmon; O'Connell, Richard J.

    2012-01-01

    Phytopathogens secrete effector proteins to manipulate their hosts for effective colonization. Hemibiotrophic fungi must maintain host viability during initial biotrophic growth and elicit host death for subsequent necrotrophic growth. To identify effectors mediating these opposing processes, we deeply sequenced the transcriptome of Colletotrichum higginsianum infecting Arabidopsis. Most effector genes are host-induced and expressed in consecutive waves associated with pathogenic transitions, indicating distinct effector suites are deployed at each stage. Using fluorescent protein tagging and transmission electron microscopy-immunogold labelling, we found effectors localised to stage-specific compartments at the host-pathogen interface. In particular, we show effectors are focally secreted from appressorial penetration pores before host invasion, revealing new levels of functional complexity for this fungal organ. Furthermore, we demonstrate that antagonistic effectors either induce or suppress plant cell death. Based on these results we conclude that hemibiotrophy in Colletotrichum is orchestrated through the coordinated expression of antagonistic effectors supporting either cell viability or cell death. PMID:22496661

  3. Highly selective biotransformation of ginsenoside Rb1 to Rd by the phytopathogenic fungus Cladosporium fulvum (syn. Fulvia fulva).

    Science.gov (United States)

    Zhao, Xuesong; Wang, Juan; Li, Jie; Fu, Ling; Gao, Juan; Du, Xiuli; Bi, Hongtao; Zhou, Yifa; Tai, Guihua

    2009-05-01

    Fourteen phytopathogenic fungi were tested for their ability to transform the major ginsenosides to the active minor ginsenoside Rd. The transformation products were identified by TLC and HPLC, and their structures were assigned by NMR analysis. Cladosporium fulvum, a tomato pathogen, was found to transform major ginsenoside Rb(1) to Rd as the sole product. The following optimum conditions for transforming Rd by C. fulvum were determined: the time of substrate addition, 24 h; substrate concentration, 0.25 mg ml(-1); temperature, 37 degrees C; pH 5.0; and biotransformation period, 8 days. At these optimum conditions, the maximum yield was 86% (molar ratio). Further, a preparative scale transformation with C. fulvum was performed at a dose of 100 mg of Rb(1) by a yield of 80%. This fungus has potential to be applied on the preparation for Rd in pharmaceutical industry.

  4. Cloning, characterization and functional expression of an endoglucanase-encoding gene from the phytopathogenic fungus Macrophomina phaseolina.

    Science.gov (United States)

    Wang, H; Jones, R W

    1995-05-26

    An endoglucanase-encoding clone (egl2) was isolated from the phytopathogenic soilborne deuteromycete fungus Macrophomina phaseolina (Mp). Clones were obtained from a cDNA library by functional expression in Escherichia coli. The egl2 clone hybridized to a 1.3-kb mRNA. Expression is induced by carboxymethylcellulose (CMC) and repressed by glucose. The deduced amino acid (aa) sequence revealed strong similarity to the egl3 from Trichoderma reesei (Tr) (72% for identical residues and 81% with conservative substitution over a span of 324 aa). The Mp egl2 lacks the cellulose-binding domain and linker region found in the Tr egl3. Different codon usage between the two fungi resulted in a much shorter span of nucleotide homology. The Egl2 protein cleaves cellodextrins with continguous beta, 1-4 linkages of four and larger, and shows activity against CMC and birchwood xylan.

  5. Identification and Antagonism Study of a Novel Chitinase-producing Bacterium Burkholderia Sp.C3 against Phytopathogenic Fungi

    Institute of Scientific and Technical Information of China (English)

    金虹; TAO Yong

    2006-01-01

    Through a modified agar well diffusion assay, antagonism of a novel chitinase-producing strain C3 against the phytopathogenic fungi including Phoma wasabiae Yokogi,Heterostrophus, Exserohilum Turcicum, Curwularia (Walk) Boed, Thantephorus cucumris, Fusarium graminearum was tested. The data showed that the crude cxtracts of strain C3 had stable antifungal activity in the range of pH 5.0 to pH 8.0. The active components were heat labile and sensitive to proteinase K. A series of experiments supported that the compound responsible for inhibitory activity appeared to be ehitinase. The 16s rDNA analysis indicated that C3 was subject to genus Burkholderia. Pbenotypic characterization of C3 was also consisted with the result of molecular identification.

  6. [Enhanced resistance to phytopathogenic bacteria in transgenic tobacco plants with synthetic gene of antimicrobial peptide cecropin P1].

    Science.gov (United States)

    Zakharchenko, N S; Rukavtsova, E B; Gudkov, A T; Bur'ianov, Ia I

    2005-11-01

    Plasmids with a synthetic gene of the mammalian antimicrobial peptide cecropin P1 (cecP1) controlled by the constitutive promoter 35S RNA of cauliflower mosaic virus were constructed. Agrobacterial transformation of tobacco plants was conducted using the obtained recombinant binary vector. The presence of gene cecP1 in the plant genome was confirmed by PCR. The expression of gene cecP1 in transgenic plants was shown by Northern blot analysis. The obtained transgenic plants exhibit enhanced resistance to phytopathogenic bacteria Pseudomonas syringae, P. marginata, and Erwinia carotovora. The ability of transgenic plants to express cecropin P1 was transmitted to the progeny. F1 and F2 plants had the normal phenotype (except for a changed coloration of flowers) and retained the ability to produce normal viable seeds upon self-pollination. Lines of F1 plants with Mendelian segregation of transgenic traits were selected.

  7. Development of SSR Markers for a Phytopathogenic Fungus, Blumeria graminis f.sp. tritici, Using a FIASCO Protocol

    Institute of Scientific and Technical Information of China (English)

    WANG Meng; XUE Fei; YANG Peng; DUAN Xia-yu; ZHOU Yi-lin; SHEN Chong-yao; ZHANG Guo-zhen; WANG Bao-tong

    2014-01-01

    Simple sequence repeats (SSR) have been widely used as molecular markers due to their abundance and high polymorphism. However, up to now, the SSR markers had not been developed in the obligate biotrophic phytopathogenic fungus, Blumeria graminis f.sp. tritici. From (AC)10 and (AG)10 enriched genomic libraries for Bgt, 25 primer pairs were designed using the FIASCO (fast isolation by AFLP of sequences containing repeats) protocol. Five primer pairs exhibited polymorphism with allelic diversity from two to seven alleles and produced 29 alleles in a survey of 90 isolates collected from six provinces (cities) in China, while the others displayed monomorphic. Levels of observed heterozygosity ranged from 0.000-0.044 (mean 0.025) and expected heterozygosity ranged from 0.297-0.816 (mean 0.538). These molecular markers provide a novel source to genetic diversity assays and to genetic and physical mapping of Bgt. SSR markers of Bgt need to be further explored.

  8. Inhibitory effect of chitosan on growth of the fungal phytopathogen, Sclerotinia sclerotiorum, and sclerotinia rot of carrot

    Institute of Scientific and Technical Information of China (English)

    WANG Qing; ZUO Jin-hua; WANG Qian; NA Yang; GAO Li-pu

    2015-01-01

    The antifungal activity of chitosan on a common fungal phytopathogen, Sclerotinia sclerotiorum, and the control effect on sclerotinia rot of carrot were investigated. Mycelial growth and fungal biomass were strongly inhibited by chitosan. Using propidium iodide stain combined with lfuorescent microscopy, the plasma membrane of chitosan-treated S. sclerotiorum mycelia was observed to be markedly damaged. Concomitantly, protein leakage and lipid peroxidation was also found to be signiifcantly higher in chitosan-treated mycelia compared to the control. Chitosan provided an effective control of sclerotinia rot of carrot, with induction of activity of defense-related enzymes including polyphenoloxidase and peroxidase. These data suggest that the effects of chitosan on sclerotinia rot of carrot may be associated with the direct damage to the plasma membrane and lipid peroxidation of S. sclerotiorum, and the elicitation of defense response in carrot.

  9. Antimicrobial Activity of Bacillus sp. Natural Isolates and Their Potential Use in the Biocontrol of Phytopathogenic Bacteria

    Directory of Open Access Journals (Sweden)

    Tanja Berić

    2012-01-01

    Full Text Available Screening of 203 Bacillus sp. natural isolates for antimicrobial activity against phytopathogenic bacteria showed that 127 tested strains inhibit at least one sensitive strain, which illustrates their potential use as biocontrol agents. Among them, 104 isolates showed significant antagonism against Xanthomonas oryzae pv. oryzae, and only one of these (VPS50.2 synthesizes bacteriocin. An additional screening tested whether 51 isolates contained genes involved in the biosynthesis of lipopeptides of the iturin and surfactin classes. Results show that 33 isolates harbour the operon for iturin biosynthesis, and six of them carry the sfp gene, responsible for the biosynthesis of surfactin. Lipopeptide purification from the supernatant of isolate SS12.9 (identified as B. subtilis or B. amyloliquefaciens was performed using ethyl acetate extraction, ultrafiltration and reversed phase HPLC. Mass spectrometry analysis confirmed that isolate SS12.9 produces a substance of the iturin class with potential for biocontrol of X. oryzae pv. oryzae.

  10. Glucanases and chitinases as causal agents in the protection of Acacia extrafloral nectar from infestation by phytopathogens.

    Science.gov (United States)

    González-Teuber, Marcia; Pozo, María J; Muck, Alexander; Svatos, Ales; Adame-Alvarez, Rosa M; Heil, Martin

    2010-03-01

    Nectars are rich in primary metabolites and attract mutualistic animals, which serve as pollinators or as an indirect defense against herbivores. Their chemical composition makes nectars prone to microbial infestation. As protective strategy, floral nectar of ornamental tobacco (Nicotiana langsdorffii x Nicotiana sanderae) contains "nectarins," proteins producing reactive oxygen species such as hydrogen peroxide. By contrast, pathogenesis-related (PR) proteins were detected in Acacia extrafloral nectar (EFN), which is secreted in the context of defensive ant-plant mutualisms. We investigated whether these PR proteins protect EFN from phytopathogens. Five sympatric species (Acacia cornigera, A. hindsii, A. collinsii, A. farnesiana, and Prosopis juliflora) were compared that differ in their ant-plant mutualism. EFN of myrmecophytes, which are obligate ant-plants that secrete EFN constitutively to nourish specialized ant inhabitants, significantly inhibited the growth of four out of six tested phytopathogenic microorganisms. By contrast, EFN of nonmyrmecophytes, which is secreted only transiently in response to herbivory, did not exhibit a detectable inhibitory activity. Combining two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis with nanoflow liquid chromatography-tandem mass spectrometry analysis confirmed that PR proteins represented over 90% of all proteins in myrmecophyte EFN. The inhibition of microbial growth was exerted by the protein fraction, but not the small metabolites of this EFN, and disappeared when nectar was heated. In-gel assays demonstrated the activity of acidic and basic chitinases in all EFNs, whereas glucanases were detected only in EFN of myrmecophytes. Our results demonstrate that PR proteins causally underlie the protection of Acacia EFN from microorganisms and that acidic and basic glucanases likely represent the most important prerequisite in this defensive function.

  11. Do induced responses mediate the ecological interactions between the specialist herbivores and phytopathogens of an alpine plant?

    Directory of Open Access Journals (Sweden)

    Gregory Röder

    Full Text Available Plants are not passive victims of the myriad attackers that rely on them for nutrition. They have a suite of physical and chemical defences, and are even able to take advantage of the enemies of their enemies. These strategies are often only deployed upon attack, so may lead to indirect interactions between herbivores and phytopathogens. In this study we test for induced responses in wild populations of an alpine plant (Adenostyles alliariae that possesses constitutive chemical defence (pyrrolizidine alkaloids and specialist natural enemies (two species of leaf beetle, Oreina elongata and Oreina cacaliae, and the phytopathogenic rust Uromyces cacaliae. Plants were induced in the field using chemical elicitors of the jasmonic acid (JA and salicylic acid (SA pathways and monitored for one month under natural conditions. There was evidence for induced resistance, with lower probability and later incidence of attack by beetles in JA-induced plants and of rust infection in SA-induced plants. We also demonstrate ecological cross-effects, with reduced fungal attack following JA-induction, and a cost of SA-induction arising from increased beetle attack. As a result, there is the potential for negative indirect effects of the beetles on the rust, while in the field the positive indirect effect of the rust on the beetles appears to be over-ridden by direct effects on plant nutritional quality. Such interactions resulting from induced susceptibility and resistance must be considered if we are to exploit plant defences for crop protection using hormone elicitors or constitutive expression. More generally, the fact that induced defences are even found in species that possess constitutively-expressed chemical defence suggests that they may be ubiquitous in higher plants.

  12. Plant carbohydrate scavenging through tonB-dependent receptors: a feature shared by phytopathogenic and aquatic bacteria.

    Science.gov (United States)

    Blanvillain, Servane; Meyer, Damien; Boulanger, Alice; Lautier, Martine; Guynet, Catherine; Denancé, Nicolas; Vasse, Jacques; Lauber, Emmanuelle; Arlat, Matthieu

    2007-02-21

    TonB-dependent receptors (TBDRs) are outer membrane proteins mainly known for the active transport of iron siderophore complexes in Gram-negative bacteria. Analysis of the genome of the phytopathogenic bacterium Xanthomonas campestris pv. campestris (Xcc), predicts 72 TBDRs. Such an overrepresentation is common in Xanthomonas species but is limited to only a small number of bacteria. Here, we show that one Xcc TBDR transports sucrose with a very high affinity, suggesting that it might be a sucrose scavenger. This TBDR acts with an inner membrane transporter, an amylosucrase and a regulator to utilize sucrose, thus defining a new type of carbohydrate utilization locus, named CUT locus, involving a TBDR for the transport of substrate across the outer membrane. This sucrose CUT locus is required for full pathogenicity on Arabidopsis, showing its importance for the adaptation to host plants. A systematic analysis of Xcc TBDR genes and a genome context survey suggested that several Xcc TBDRs belong to other CUT loci involved in the utilization of various plant carbohydrates. Interestingly, several Xcc TBDRs and CUT loci are conserved in aquatic bacteria such as Caulobacter crescentus, Colwellia psychrerythraea, Saccharophagus degradans, Shewanella spp., Sphingomonas spp. or Pseudoalteromonas spp., which share the ability to degrade a wide variety of complex carbohydrates and display TBDR overrepresentation. We therefore propose that TBDR overrepresentation and the presence of CUT loci designate the ability to scavenge carbohydrates. Thus CUT loci, which seem to participate to the adaptation of phytopathogenic bacteria to their host plants, might also play a very important role in the biogeochemical cycling of plant-derived nutrients in marine environments. Moreover, the TBDRs and CUT loci identified in this study are clearly different from those characterized in the human gut symbiont Bacteroides thetaiotaomicron, which allow glycan foraging, suggesting a convergent

  13. Plant carbohydrate scavenging through tonB-dependent receptors: a feature shared by phytopathogenic and aquatic bacteria.

    Directory of Open Access Journals (Sweden)

    Servane Blanvillain

    Full Text Available TonB-dependent receptors (TBDRs are outer membrane proteins mainly known for the active transport of iron siderophore complexes in Gram-negative bacteria. Analysis of the genome of the phytopathogenic bacterium Xanthomonas campestris pv. campestris (Xcc, predicts 72 TBDRs. Such an overrepresentation is common in Xanthomonas species but is limited to only a small number of bacteria. Here, we show that one Xcc TBDR transports sucrose with a very high affinity, suggesting that it might be a sucrose scavenger. This TBDR acts with an inner membrane transporter, an amylosucrase and a regulator to utilize sucrose, thus defining a new type of carbohydrate utilization locus, named CUT locus, involving a TBDR for the transport of substrate across the outer membrane. This sucrose CUT locus is required for full pathogenicity on Arabidopsis, showing its importance for the adaptation to host plants. A systematic analysis of Xcc TBDR genes and a genome context survey suggested that several Xcc TBDRs belong to other CUT loci involved in the utilization of various plant carbohydrates. Interestingly, several Xcc TBDRs and CUT loci are conserved in aquatic bacteria such as Caulobacter crescentus, Colwellia psychrerythraea, Saccharophagus degradans, Shewanella spp., Sphingomonas spp. or Pseudoalteromonas spp., which share the ability to degrade a wide variety of complex carbohydrates and display TBDR overrepresentation. We therefore propose that TBDR overrepresentation and the presence of CUT loci designate the ability to scavenge carbohydrates. Thus CUT loci, which seem to participate to the adaptation of phytopathogenic bacteria to their host plants, might also play a very important role in the biogeochemical cycling of plant-derived nutrients in marine environments. Moreover, the TBDRs and CUT loci identified in this study are clearly different from those characterized in the human gut symbiont Bacteroides thetaiotaomicron, which allow glycan foraging

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

    Science.gov (United States)

    Martínez-García, Pedro Manuel; Ruano-Rosa, David; Schilirò, Elisabetta; Prieto, Pilar; Ramos, Cayo; Rodríguez-Palenzuela, Pablo; Mercado-Blanco, Jesús

    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.

  15. Effect of Verticillium lecanii on biological characteristics and life table of Serangium japonicum (Coleoptera: Coccinellidae), a predator of whiteflies under laboratory conditions

    Institute of Scientific and Technical Information of China (English)

    Lazreg Fatiha; Zhen Huang; Shun-Xiang Ren; Shaukat Ali

    2008-01-01

    Effects of entomopathogenic fungus Verticillium lecanii on biological characteristics and life table of Serangiumjaponicum, a predator of whiteflies against five different conidial concentrations (1×104, 1×105, 1×106, 1×107, and 1×108 conidia/mL) were studied under laboratory conditions. The developmental periods for all immature stages (from eggs, 1st, 2nd, 3rd, 4th instar nymph and pupae up to emergence) among the treatments were significantly different when compared to that of control, and the longest development period was observed as treated with 1×108spore/mL. However, no significant difference on the percent survival of all immature stages was observed among the treatments and control. Also, there were no significantly different effects of V. lecanii on mean generation time, intrinsic rate, the finite rate of increase and longevity of S. japonicum among the treatments and control.

  16. Isolation, Identification and Activity of an Antagonistic Spore Bacterium against Cotton Verticillium Wilt%一株棉花黄萎病拮抗芽孢细菌的分离鉴定及活性检测

    Institute of Scientific and Technical Information of China (English)

    张冬冬; 李术娜; 郭晓军; 朱宝成

    2012-01-01

    采用改良的琼脂平板扩散法,通过初筛和复筛,得到一株对棉花黄萎病病原菌大丽轮枝菌(Verticillium dahliae)具有较强的抑菌活性的拮抗细菌Z-5菌株.通过对该菌株的形态特征观察、生理生化鉴定和16S rDNA 的序列分析,确定拮抗细菌Z-5菌株为Bacillus malacitensis.盆栽实验表明Z-5菌株对大丽轮枝菌的防治效果达到76.05%,对棉花黄萎病具有明显的防治作用.%A bacterial strain called Z-5 that had strong inhibition capability against Verticillium dahliae V-190 was obtained via preliminary and secondary screening with the improved agar plate diffusion method. Characterized by morphological and cultural features observation, physiological and biochemical experiments, and 16S rDNA sequence analysis, the Z-5 strain was initially identified as Bacillus malacitensis. Control of cotton Verticillium wilt by this antagonistic bacterium was assessed via potted plants. Results showed that the control efficacy of the antagonistic Z-5 strain on cotton Verticillium wilt was 76.05%. Preliminary results demonstrated that the antagonistic Z-5 bacterial strain could significantly inhibit the growth of V. dahliae.

  17. Application and bioactive properties of CaTI, a trypsin inhibitor from Capsicum annuum seeds: membrane permeabilization, oxidative stress and intracellular target in phytopathogenic fungi cells.

    Science.gov (United States)

    Silva, Marciele S; Ribeiro, Suzanna Ff; Taveira, Gabriel B; Rodrigues, Rosana; Fernandes, Katia Vs; Carvalho, André O; Vasconcelos, Ilka Maria; Mello, Erica Oliveira; Gomes, Valdirene M

    2017-08-01

    During the last few years, a growing number of antimicrobial peptides have been isolated from plants and particularly from seeds. Recent results from our laboratory have shown the purification of a new trypsin inhibitor, named CaTI, from chilli pepper (Capsicum annuum L.) seeds. This study aims to evaluate the antifungal activity and mechanism of action of CaTI on phytopathogenic fungi and detect the presence of protease inhibitors in other species of this genus. Our results show that CaTI can inhibit the growth of the phytopathogenic fungi Colletotrichum gloeosporioides and C. lindemuthianum. CaTI can also permeabilize the membrane of all tested fungi. When testing the inhibitor on its ability to induce reactive oxygen species, an induction of reactive oxygen species (ROS) and nitric oxide (NO) particularly in Fusarium species was observed. Using CaTI coupled to fluorescein isothiocyanate (FITC), it was possible to determine the presence of the inhibitor inside the hyphae of the Fusarium oxysporum fungus. The search for protease inhibitors in other Capsicum species revealed their presence in all tested species. This paper shows the antifungal activity of protease inhibitors such as CaTI against phytopathogenic fungi. Antimicrobial peptides, among which the trypsin protease inhibitor family stands out, are present in different species of the genus Capsicum and are part of the chemical arsenal that plants use to defend themselves against pathogens. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  18. Expression of antimicrobial peptides thanatin(S) in transgenic Arabidopsis enhanced resistance to phytopathogenic fungi and bacteria.

    Science.gov (United States)

    Wu, Tingquan; Tang, Dingzhong; Chen, Weida; Huang, Hexun; Wang, Rui; Chen, Yongfang

    2013-09-15

    Thanatin(S) is an analog of thanatin, an insect antimicrobial peptide possessing strong and broad spectrum of antimicrobial activity. In order to investigate if the thanatin could be used in engineering transgenic plants for increased resistance against phytopathogens, the synthetic thanatin(S) was introduced into Arabidopsis thaliana plants. To increase the expression level of thanatin(S) in plants, the coding sequence was optimized by plant-preference codon. To avoid cellular protease degradation, signal peptide of rice Cht1 was fused to N terminal of thanatin(S) for secreting the expressed thanatin(S) into intercellular spaces. To evaluate the application value of thanatin(S) in plant disease control, the synthesized coding sequence of Cht1 signal peptide (Cht1SP)-thanatin(S) was ligated to plant gateway destination binary vectors pGWB11 (with FLAG tag). Meanwhile, in order to observe the subcellular localization of Cht1SP-thanatin(S)-GFP and thanatin(S)-GFP, the sequences of Cht1SP-thanatin(S) and thanatin(S) were respectively linked to pGWB5 (with GFP tag). The constructs were transformed into Arabidopsis ecotype Col-0 and mutant pad4-1 via Agrobacterium-mediated transformation. The transformants with Cht1SP-thanatin(S)-FLAG fusion gene were analyzed by genomic PCR, real-time PCR, and western blots and the transgenic Arabidopsis plants introduced respectively Cht1SP-thanatin(S)-GFP and thanatin(S)-GFP were observed by confocal microscopy. Transgenic plants expressing Cht1SP-thanatin(S)-FLAG fusion protein showed antifungal activity against Botrytis cinerea and powdery mildew, as well as antibacterial activity against Pseudomonas syringae pv. tomato. And the results from confocal observation showed that the GFP signal from Cht1SP-thanatin(S)-GFP transgenic Arabidopsis plants occurred mainly in intercellular space, while that from thanatin(S)-GFP transgenic plants was mainly detected in the cytoplasm and that from empty vector transgenic plants was distributed

  19. 大丽轮枝菌(Verticillium dahliae VdLs.17)分泌组预测及分析%Prediction and Analysis of Verticillium dahliae VdLs.17 Secretome

    Institute of Scientific and Technical Information of China (English)

    田李; 陈捷胤; 陈相永; 汪佳妮; 戴小枫

    2011-01-01

    [Objective] The aim of this study was to predict and analyze secretome of Verticillium dahliae, an important soil-born fungal pathogen that causes vascular wilt diseases. [Method] The softwares SignalP, TargetP, TMHMM, Big-pi and PROSITE were used to predict the sectetome of V. Dahliae. Protein databases were used for secretome annotation to find putative pectinase, cellulose and pathogen-host interaction proteins. The proteins with common fungal effector character were investigated by a set of computer algorithms. By using BLASTP, the secretome of V. Dahliae and V. Albo-atrum was compared to find V. Dahliae specifically secreted proteins. [Result] A total of 922 possible secreted proteins were identified in V. Dahliae scretome. The most frequent amino acid in signal peptides is alanine, and the least is glutamic acid and aspartic acid. The amino acid in the position -3 and -1 are very conserved. The secretome is equipped with 158 carbohydrate-active enzymes which include 10 pectic hydrolases and 14 pectic lyases, 190 putative pathogen-host interaction proteins, 97 RxLx[EDQ] motif containing proteins and 52 small cysteine-rich secreted proteins, and 58 V. Dahliae specifically secreted proteins compared with V. Albo-atrum. [Conclusion] Thisstudy has provided secretome prediction algorithms for V. Dahliae. Length of signal peptides vary greatly. Composition of signal peptides are mainly aliphatic amino acid and sequences of signal peptides near C-terminal are conservative. V. Dahliae secretome are rich in potential pectinase, pathogen-host interaction protein, RxLx[EDQ] motif containing protein and small cysteine-rich protein which may play roles in V. Dahliae pathogenetic mechanism.%[目的]预测并分析大丽轮枝菌基因组范围内的分泌蛋白,为大丽轮枝菌分泌蛋白致病机理的研究奠定基础.[方法]利用已公布的大丽轮枝菌全基因组序列,组合使用生物信息学软件SignalP、TargetP、TMHMM、Big-pi和PROSITE,预测大丽

  20. Transgenic Brassica juncea plants expressing MsrA1, a synthetic cationic antimicrobial peptide, exhibit resistance to fungal phytopathogens.

    Science.gov (United States)

    Rustagi, Anjana; Kumar, Deepak; Shekhar, Shashi; Yusuf, Mohd Aslam; Misra, Santosh; Sarin, Neera Bhalla

    2014-06-01

    Cationic antimicrobial peptides (CAPs) have shown potential against broad spectrum of phytopathogens. Synthetic versions with desirable properties have been modeled on these natural peptides. MsrA1 is a synthetic chimera of cecropin A and melittin CAPs with antimicrobial properties. We generated transgenic Brassica juncea plants expressing the msrA1 gene aimed at conferring fungal resistance. Five independent transgenic lines were evaluated for resistance to Alternaria brassicae and Sclerotinia sclerotiorum, two of the most devastating pathogens of B. juncea crops. In vitro assays showed inhibition by MsrA1 of Alternaria hyphae growth by 44-62 %. As assessed by the number and size of lesions and time taken for complete leaf necrosis, the Alternaria infection was delayed and restricted in the transgenic plants with the protection varying from 69 to 85 % in different transgenic lines. In case of S. sclerotiorum infection, the lesions were more severe and spread profusely in untransformed control compared with transgenic plants. The sclerotia formed in the stem of untransformed control plants were significantly more in number and larger in size than those present in the transgenic plants where disease protection of 56-71.5 % was obtained. We discuss the potential of engineering broad spectrum biotic stress tolerance by transgenic expression of CAPs in crop plants.

  1. Chitinase genes revealed and compared in bacterial isolates, DNA extracts and a metagenomic library from a phytopathogen suppressive soil

    Energy Technology Data Exchange (ETDEWEB)

    Hjort, K.; Bergstrom, M.; Adesina, M.F.; Jansson, J.K.; Smalla, K.; Sjoling, S.

    2009-09-01

    Soil that is suppressive to disease caused by fungal pathogens is an interesting source to target for novel chitinases that might be contributing towards disease suppression. In this study we screened for chitinase genes, in a phytopathogen-suppressive soil in three ways: (1) from a metagenomic library constructed from microbial cells extracted from soil, (2) from directly extracted DNA and (3) from bacterial isolates with antifungal and chitinase activities. Terminal-restriction fragment length polymorphism (T-RFLP) of chitinase genes revealed differences in amplified chitinase genes from the metagenomic library and the directly extracted DNA, but approximately 40% of the identified chitinase terminal-restriction fragments (TRFs) were found in both sources. All of the chitinase TRFs from the isolates were matched to TRFs in the directly extracted DNA and the metagenomic library. The most abundant chitinase TRF in the soil DNA and the metagenomic library corresponded to the TRF{sup 103} of the isolate, Streptomyces mutomycini and/or Streptomyces clavifer. There were good matches between T-RFLP profiles of chitinase gene fragments obtained from different sources of DNA. However, there were also differences in both the chitinase and the 16S rRNA gene T-RFLP patterns depending on the source of DNA, emphasizing the lack of complete coverage of the gene diversity by any of the approaches used.

  2. The transition from a phytopathogenic smut ancestor to an anamorphic biocontrol agent deciphered by comparative whole-genome analysis.

    Science.gov (United States)

    Lefebvre, François; Joly, David L; Labbé, Caroline; Teichmann, Beate; Linning, Rob; Belzile, François; Bakkeren, Guus; Bélanger, Richard R

    2013-06-01

    Pseudozyma flocculosa is related to the model plant pathogen Ustilago maydis yet is not a phytopathogen but rather a biocontrol agent of powdery mildews; this relationship makes it unique for the study of the evolution of plant pathogenicity factors. The P. flocculosa genome of ~23 Mb includes 6877 predicted protein coding genes. Genome features, including hallmarks of pathogenicity, are very similar in P. flocculosa and U. maydis, Sporisorium reilianum, and Ustilago hordei. Furthermore, P. flocculosa, a strict anamorph, revealed conserved and seemingly intact mating-type and meiosis loci typical of Ustilaginales. By contrast, we observed the loss of a specific subset of candidate secreted effector proteins reported to influence virulence in U. maydis as the singular divergence that could explain its nonpathogenic nature. These results suggest that P. flocculosa could have once been a virulent smut fungus that lost the specific effectors necessary for host compatibility. Interestingly, the biocontrol agent appears to have acquired genes encoding secreted proteins not found in the compared Ustilaginales, including necrosis-inducing-Phytophthora-protein- and Lysin-motif- containing proteins believed to have direct relevance to its lifestyle. The genome sequence should contribute to new insights into the subtle genetic differences that can lead to drastic changes in fungal pathogen lifestyles.

  3. Multimodal Generally Recognized as Safe ZnO/Nanocopper Composite: A Novel Antimicrobial Material for the Management of Citrus Phytopathogens.

    Science.gov (United States)

    Young, Mikaeel; Ozcan, Ali; Myers, Monty E; Johnson, Evan G; Graham, James H; Santra, Swadeshmukul

    2017-09-05

    Copper (Cu) bactericides/fungicides are used extensively for crop protection in agriculture. Concerns for Cu accumulation in soil, Cu leaching into the surrounding ecosystem, and development of Cu resistance in phytopathogenic bacteria are evident. While there is no suitable alternative to Cu available to date for agricultural uses, it is possible to reduce Cu per application by supplementing with Zn and improving Cu bioavailability using nanotechnology. We have prepared a non-phytotoxic composite material consisting of generally recognized as safe ZnO 800 particles and nanocopper-loaded silica gel (ZnO-nCuSi). The morphology of the ZnO-nCuSi material was characterized using scanning electron microscopy, showing ZnO particles dispersed in the silica gel matrix. ZnO-nCuSi demonstrated strong in vitro antimicrobial properties against several model plant bacterial species. Two consecutive year field efficacy results showed that agri-grade ZnO-nCuSi was effective in controlling citrus canker disease at less than half the metallic rate of the commercial cuprous oxide/zinc oxide pesticide.

  4. Marine Actinobacteria as a source of compounds for phytopathogen control: An integrative metabolic-profiling / bioactivity and taxonomical approach.

    Science.gov (United States)

    Betancur, Luz A; Naranjo-Gaybor, Sandra J; Vinchira-Villarraga, Diana M; Moreno-Sarmiento, Nubia C; Maldonado, Luis A; Suarez-Moreno, Zulma R; Acosta-González, Alejandro; Padilla-Gonzalez, Gillermo F; Puyana, Mónica; Castellanos, Leonardo; Ramos, Freddy A

    2017-01-01

    Marine bacteria are considered as promising sources for the discovery of novel biologically active compounds. In this study, samples of sediment, invertebrate and algae were collected from the Providencia and Santa Catalina coral reef (Colombian Caribbean Sea) with the aim of isolating Actinobateria-like strain able to produce antimicrobial and quorum quenching compounds against pathogens. Several approaches were used to select actinobacterial isolates, obtaining 203 strains from all samples. According to their 16S rRNA gene sequencing, a total of 24 strains was classified within Actinobacteria represented by three genera: Streptomyces, Micromonospora, and Gordonia. In order to assess their metabolic profiles, the actinobacterial strains were grown in liquid cultures, and LC-MS-based analyses from ethyl acetate fractions were performed. Based on taxonomical classification, screening information of activity against phytopathogenic strains and quorum quenching activity, as well as metabolic profiling, six out of the 24 isolates were selected for follow-up with chemical isolation and structure identification analyses of putative metabolites involved in antimicrobial activities.

  5. Sequential delivery of host-induced virulence effectors by appressoria and intracellular hyphae of the phytopathogen Colletotrichum higginsianum.

    Directory of Open Access Journals (Sweden)

    Jochen Kleemann

    Full Text Available Phytopathogens secrete effector proteins to manipulate their hosts for effective colonization. Hemibiotrophic fungi must maintain host viability during initial biotrophic growth and elicit host death for subsequent necrotrophic growth. To identify effectors mediating these opposing processes, we deeply sequenced the transcriptome of Colletotrichum higginsianum infecting Arabidopsis. Most effector genes are host-induced and expressed in consecutive waves associated with pathogenic transitions, indicating distinct effector suites are deployed at each stage. Using fluorescent protein tagging and transmission electron microscopy-immunogold labelling, we found effectors localised to stage-specific compartments at the host-pathogen interface. In particular, we show effectors are focally secreted from appressorial penetration pores before host invasion, revealing new levels of functional complexity for this fungal organ. Furthermore, we demonstrate that antagonistic effectors either induce or suppress plant cell death. Based on these results we conclude that hemibiotrophy in Colletotrichum is orchestrated through the coordinated expression of antagonistic effectors supporting either cell viability or cell death.

  6. Identification of a Polyketide Synthase Gene in the Synthesis of Phleichrome of the Phytopathogenic Fungus Cladosporium phlei.

    Science.gov (United States)

    So, Kum-Kang; Chung, Yun-Jo; Kim, Jung-Mi; Kim, Beom-Tae; Park, Seung-Moon; Kim, Dae-Hyuk

    2015-12-01

    Phleichrome, a pigment produced by the phytopathogenic fungus Cladosporium phlei, is a fungal perylenequinone whose photodynamic activity has been studied intensively. To determine the biological function of phleichrome and to engineer a strain with enhanced production of phleichrome, we identified the gene responsible for the synthesis of phleichrome. Structural comparison of phleichrome with other fungal perylenequinones suggested that phleichrome is synthesized via polyketide pathway. We recently identified four different polyketide synthase (PKS) genes encompassing three major clades of fungal PKSs that differ with respect to reducing conditions for the polyketide product. Based on in silico analysis of cloned genes, we hypothesized that the non-reducing PKS gene, Cppks1, is involved in phleichrome biosynthesis. Increased accumulation of Cppks1 transcript was observed in response to supplementation with the application of synthetic inducer cyclo-(l-Pro-l-Phe). In addition, heterologous expression of the Cppks1 gene in Cryphonectria parasitica resulted in the production of phleichrome. These results provide convincing evidence that the Cppks1 gene is responsible for the biosynthesis of phleichrome.

  7. Marine Actinobacteria as a source of compounds for phytopathogen control: An integrative metabolic-profiling / bioactivity and taxonomical approach

    Science.gov (United States)

    Betancur, Luz A.; Naranjo-Gaybor, Sandra J.; Vinchira-Villarraga, Diana M.; Moreno-Sarmiento, Nubia C.; Maldonado, Luis A.; Suarez-Moreno, Zulma R.; Acosta-González, Alejandro; Padilla-Gonzalez, Gillermo F.; Puyana, Mónica; Castellanos, Leonardo; Ramos, Freddy A.

    2017-01-01

    Marine bacteria are considered as promising sources for the discovery of novel biologically active compounds. In this study, samples of sediment, invertebrate and algae were collected from the Providencia and Santa Catalina coral reef (Colombian Caribbean Sea) with the aim of isolating Actinobateria-like strain able to produce antimicrobial and quorum quenching compounds against pathogens. Several approaches were used to select actinobacterial isolates, obtaining 203 strains from all samples. According to their 16S rRNA gene sequencing, a total of 24 strains was classified within Actinobacteria represented by three genera: Streptomyces, Micromonospora, and Gordonia. In order to assess their metabolic profiles, the actinobacterial strains were grown in liquid cultures, and LC-MS-based analyses from ethyl acetate fractions were performed. Based on taxonomical classification, screening information of activity against phytopathogenic strains and quorum quenching activity, as well as metabolic profiling, six out of the 24 isolates were selected for follow-up with chemical isolation and structure identification analyses of putative metabolites involved in antimicrobial activities. PMID:28225766

  8. Study of bio-fabrication of iron nanoparticles and their fungicidal property against phytopathogens of apple orchards.

    Science.gov (United States)

    Ahmad, Hilal; Rajagopal, Kalyanaraman; Shah, Ashiq Hussain; Bhat, Arif Hussain; Venugopal, Kalyanaraman

    2017-04-01

    Current research trends on iron nanoparticles (FeNPs) are extensively focused because of their unique magnetic and electrical properties mostly applicable in essential medical devices. However, their fungicidal property against plant pathogens is very less known until date. Present study demonstrates a green technique for blending of FeNPs by utilising aqueous extract of neem leaf (Azadirachta indica A. Juss.) as reducing agent. Various characterisation techniques such as ultraviolet (UV)-visible spectroscopy, Fourier transform infrared spectroscopy transmission electron microscopy, scanning electron microscopy and X-ray diffraction were performed for FeNPs. The authors' results demonstrate the more cluster formation of FeNPs with size distribution of 20-80 nm. The bio-fabricated FeNPs showed enhanced biocidal activity against economically important phytopathogens of apple such as Alternaria mali, Botryosphaeria dothidea and Diplodia seriata. From the obtained results, it can be suggested that further delve into green synthesis of FeNPs can address future biotechnology concerns to limit the synthesis of FeNPs by conventional methods. Furthermore, the field study on pathogenic fungi can be an effective step to verify their agricultural applications.

  9. Antimicrobial activities of novel mannosyl lipids isolated from the biocontrol fungus Simplicillium lamellicola BCP against phytopathogenic bacteria.

    Science.gov (United States)

    Le Dang, Quang; Shin, Teak Soo; Park, Myung Soo; Choi, Yong Ho; Choi, Gyung Ja; Jang, Kyoung Soo; Kim, In Seon; Kim, Jin-Cheol

    2014-04-16

    The antagonistic fungus Simplicillium lamellicola BCP has been developed as a microbial biopesticide that effectively controls the development of various plant diseases caused by both pathogenic bacteria and pathogenic fungi. Antibacterial bioassay-directed fractionation was used to isolate mannosyl lipids from S. lamellicola BCP, and the structures of these compounds were elucidated using spectral analysis and chemical degradation. Three novel mannosyl lipids were characterized and identified as halymecins F and G and (3R,5R)-3-O-β-D-mannosyl-3,5-dihydrodecanoic acid. Massoia lactone and (3R, 5R)-3-hydroxydecan-5-olide were also isolated from S. lamellicola BCP. The three novel compounds inhibited the growth of the majority of phytopathogenic bacteria that were tested, and halymecin F displayed the strongest antibacterial activity. Agrobacterium tumefaciens was the most sensitive to the three novel compounds, with IC₅₀ values ranging from 1.58 to 24.8 μg/mL. The ethyl acetate extract of the fermentation broth from the antagonistic fungus effectively reduced the bacterial wilt caused by Ralstonia solanacearum on tomato seedlings. These results indicate that S. lamellicola BCP suppresses the development of plant bacterial diseases through the production of antibacterial metabolites.

  10. Synthesis and biological evaluation of novel fluorine-containing stilbene derivatives as fungicidal agents against phytopathogenic fungi.

    Science.gov (United States)

    Jian, Weilin; He, Daohang; Xi, Pinggen; Li, Xinwei

    2015-11-18

    The rising development of resistance to conventional fungicides is driving the search for new alternative candidates to control plant diseases. In this study, a series of new fluorine-containing stilbene derivatives was synthesized on the basis of our previous quantitative structure-activity relationship analysis results. Bioassays in vivo revealed that the title compounds exhibited potent fungicidal activities against phytopathogenic fungi (Colletotrichum lagenarium and Pseudoperonospora cubensis) from cucumber plants. In comparison to the previous results, the introduction of a fluorine moiety showed improved activities of some compounds against those fungi. Notably, compound 9 exhibited a control efficacy against C. lagenarium (83.4 ± 1.3%) comparable to that of commercial fungicide (82.7 ± 1.7%). For further understanding the possible mode of action of the stilbene against C. lagenarium, the effects on hyphal morphology, electrolyte leakage, and respiration of mycelial cell suspension were studied. Microscopic observation showed considerably deformed mycelial morphology. The conductivity of mycelial suspension increased in the presence of compound 9, whereas no significantly inhibitory effect on respiration was observed. Taken together, the fungicidal mechanism of this stilbene is associated with its membrane disruption effect, resulting in increased membrane permeability. These results provide important clues for mechanistic study and derivatization of stilbenes as alternative sources of fungicidal agents for plant disease control.

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

    Science.gov (United States)

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

    2014-07-01

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

  12. Crystallographic structure and substrate-binding interactions of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri.

    Science.gov (United States)

    Balan, Andrea; Santacruz-Pérez, Carolina; Moutran, Alexandre; Ferreira, Luís Carlos Souza; Neshich, Goran; Gonçalves Barbosa, João Alexandre Ribeiro

    2008-02-01

    In Xanthomonas axonopodis pv. citri (Xac or X. citri), the modA gene codes for a periplasmic protein (ModA) that is capable of binding molybdate and tungstate as part of the ABC-type transporter required for the uptake of micronutrients. In this study, we report the crystallographic structure of the Xac ModA protein with bound molybdate. The Xac ModA structure is similar to orthologs with known three-dimensional structures and consists of two nearly symmetrical domains separated by a hinge region where the oxyanion-binding site lies. Phylogenetic analysis of different ModA orthologs based on sequence alignments revealed three groups of molybdate-binding proteins: bacterial phytopathogens, enterobacteria and soil bacteria. Even though the ModA orthologs are segregated into different groups, the ligand-binding hydrogen bonds are mostly conserved, except for Archaeglobus fulgidus ModA. A detailed discussion of hydrophobic interactions in the active site is presented and two new residues, Ala38 and Ser151, are shown to be part of the ligand-binding pocket.

  13. The Ve-mediated resistance response of the tomato to Verticillium dahliae involves H2O2, peroxidase and lignins and drives PAL gene expression

    Directory of Open Access Journals (Sweden)

    Merino Fuencisla

    2010-10-01

    Full Text Available Abstract Background Verticillium dahliae is a fungal pathogen that infects a wide range of hosts. The only known genes for resistance to Verticillium in the Solanaceae are found in the tomato (Solanum lycopersicum Ve locus, formed by two linked genes, Ve1 and Ve2. To characterize the resistance response mediated by the tomato Ve gene, we inoculated two nearly isogenic tomato lines, LA3030 (ve/ve and LA3038 (Ve/Ve, with V. dahliae. Results We found induction of H2O2 production in roots of inoculated plants, followed by an increase in peroxidase activity only in roots of inoculated resistant plants. Phenylalanine-ammonia lyase (PAL activity was also increased in resistant roots 2 hours after inoculation, while induction of PAL activity in susceptible roots was not seen until 48 hours after inoculation. Phenylpropanoid metabolism was also affected, with increases in ferulic acid, p-coumaric acid, vanillin and p-hydroxybenzaldehyde contents in resistant roots after inoculation. Six tomato PAL cDNA sequences (PAL1 - PAL6 were found in the SolGenes tomato EST database. RT-PCR analysis showed that these genes were expressed in all organs of the plant, albeit at different levels. Real-time RT-PCR indicated distinct patterns of expression of the different PAL genes in V. dahliae-inoculated roots. Phylogenetic analysis of 48 partial PAL cDNAs corresponding to 19 plant species grouped angiosperm PAL sequences into four clusters, suggesting functional differences among the six tomato genes, with PAL2 and PAL6 presumably involved in lignification, and the remaining PAL genes implicated in other biological processes. An increase in the synthesis of lignins was found 16 and 28 days after inoculation in both lines; this increase was greater and faster to develop in the resistant line. In both resistant and susceptible inoculated plants, an increase in the ratio of guaiacyl/syringyl units was detected 16 days after inoculation, resulting from the lowered amount

  14. Molecular evidence for the involvement of a polygalacturonase-inhibiting protein, GhPGIP1, in enhanced resistance to Verticillium and Fusarium wilts in cotton

    Science.gov (United States)

    Liu, Nana; Zhang, Xueyan; Sun, Yun; Wang, Ping; Li, Xiancai; Pei, Yakun; Li, Fuguang; Hou, Yuxia

    2017-01-01

    Polygalacturonase-inhibiting protein (PGIP), belonging to a group of plant defence proteins, specifically inhibits endopolygalacturonases secreted by pathogens. Herein, we showed that purified GhPGIP1 is a functional inhibitor of Verticillium dahliae and Fusarium oxysporum f. sp. vasinfectum, the two fungal pathogens causing cotton wilt. Transcription of GhPGIP1 was increased in cotton upon infection, wounding, and treatment with defence hormone and H2O2. Resistance by GhPGIP1 was examined by its virus-induced gene silencing in cotton and overexpression in Arabidopsis. GhPGIP1-silenced cotton was highly susceptible to the infections. GhPGIP1 overexpression in transgenic Arabidopsis conferred resistance to the infection, accompanied by enhanced expression of pathogenesis-related proteins (PRs), isochorismate synthase 1 (ICS1), enhanced disease susceptibility 1 (EDS1), and phytoalexin-deficient 4 (PAD4) genes. Transmission electron microscopy revealed cell wall alteration and cell disintegration in plants inoculated with polygalacturonase (PGs), implying its role in damaging the cell wall. Docking studies showed that GhPGIP1 interacted strongly with C-terminal of V. dahliae PG1 (VdPG1) beyond the active site but weakly interacted with C-terminal of F. oxysporum f. sp. vasinfectum (FovPG1). These findings will contribute towards the understanding of the roles of PGIPs and in screening potential combat proteins with novel recognition specificities against evolving pathogenic factors for countering pathogen invasion. PMID:28079053

  15. Isolation and identification of the antagonistic strain DM-54 of Bacillus amyloliquefacien against Verticillium dahliae, and optimization of antifungal protein producing conditions

    Institute of Scientific and Technical Information of China (English)

    Ying ZHU; Shuna LI; Hongshui YUAN; Xiaojun GUO; Baocheng ZHU

    2009-01-01

    The strains capable of resistance against Verticillium dahliae Kleb were isolated and screened from the soils of cotton fields from several different provinces in China. A strain, coded DM-54, with a rather high antagonistic activity was obtained. Its morphological characteristics, physiological and biochemical properties and a 16 S rDNA sequence of this strain were further studied. The DM-54 strain was finally identified as a kind of Bacillus amyloliquefacien. Through a single factor experiment and an orthogonal experiment, the optimal shaking flask fermentation condition of strain DM-54 was found to be: media composed of 5% dextrin, 3% soy peptone, 0.02% MgSO4 0.01% CaCl2, initial pH 7.0 and 10% inoculum volume, media volume 30/250 (mL/mL), fermentation temperature at 32℃, rotating speed activity was distinguished to be elevated, at about 39.9%. Our research offers an effective means for the massive production of antagonistic proteins.

  16. Assessment of genetic diversity among Chinese upland cottons with Fusarium and/or Verticillium wilts resistance by AFLP and SSR markers

    Institute of Scientific and Technical Information of China (English)

    WANG Xingfen; MA Jun; YANG Shuo; ZHANG Guiyin; MA Zhiying

    2007-01-01

    Genetic diversity among 95 Chinese upland cottons with Fusarium and/or Verticillium wilts resistance was estimated using Amplified Fragment Length Polymorphism (AFLP) and Simple Sequence Repeat (SSR) markers.Twenty EcoRI-MseI AFLP and 19 SSR primers with polymorphism were selected to perform the fingerprinting.The results showed that 20 AFLP primer pairs produced a total of 1 480 major bands among 95 genotypes,and 214 were polymorphic bands.The number of total bands per primer pair ranged from 47 to 109,with an average of 74.0.The polymorphism information content (PIC) values for the 20 primer pairs varied from 0.01 (E-ACT/M-CAT) to 0.24 (E-ACA/MCTA),and the average value was 0.09.Nineteen SSR primers generated 89 DNA bands,of which 61 were polymorphic.The total number of alleles per locus varied from 3 to 8,with an average of 4.7.The average PIC value for the SSR amplification products was 0.69.Genetic similarity estimates for the entire data set ranged from 0.978 to 0.998 based on AFLP and SSR bands.It was proved that the close genetic relationship and narrow genetic diversity existed in the tested cultivars.The clustering patterns could not be correlated to the geographic origin,the pedigree and common parentage of the cultivars.

  17. 山西棉田黄萎病菌致病力差异研究%Research on Virulence Difference of Verticillium Wilt in Shanxi

    Institute of Scientific and Technical Information of China (English)

    许爱玲; 段国琪; 张战备; 沈红星; 张开峰; 解广田

    2013-01-01

    In the same piece of cotton field, different resistance transgenic cotton varieties are planted. Using choice function of different resistant cotton varieties to Verticillium wilt bacteria, Vertkillium wilt strains had been gotten from different breed separations; In the greenhouse, using two different obvious disease resistance varieties pathogenic force were measured to determine whether there are different pathogenic force strains in the same piece of cotton field.%在同一块棉田中种植不同抗病性的转基因抗虫棉,利用不同抗性的棉花品种对棉花黄萎病菌的选择作用,从不同品种上分离获得不同的黄萎病菌系,并在温室中采用2个抗病性差异明显的品种对其致病力进行测定,以确定同一块棉田中是否存在不同致病力菌系.结果表明,不同抗病性品种上分离获得的病原菌致病力存在显著差异,同一块棉田中存在各种致病力菌系.

  18. Influence of Soil Type, Cultivar and Verticillium dahliae on the Structure of the Root and Rhizosphere Soil Fungal Microbiome of Strawberry

    Science.gov (United States)

    Nallanchakravarthula, Srivathsa; Mahmood, Shahid; Alström, Sadhna; Finlay, Roger D.

    2014-01-01

    Sustainable management of crop productivity and health necessitates improved understanding of the ways in which rhizosphere microbial populations interact with each other, with plant roots and their abiotic environment. In this study we examined the effects of different soils and cultivars, and the presence of a soil-borne fungal pathogen, Verticillium dahliae, on the fungal microbiome of the rhizosphere soil and roots of strawberry plants, using high-throughput pyrosequencing. Fungal communities of the roots of two cultivars, Honeoye and Florence, were statistically distinct from those in the rhizosphere soil of the same plants, with little overlap. Roots of plants growing in two contrasting field soils had high relative abundance of Leptodontidium sp. C2 BESC 319 g whereas rhizosphere soil was characterised by high relative abundance of Trichosporon dulcitum or Cryptococcus terreus, depending upon the soil type. Differences between different cultivars were not as clear. Inoculation with the pathogen V. dahliae had a significant influence on community structure, generally decreasing the number of rhizosphere soil- and root-inhabiting fungi. Leptodontidium sp. C2 BESC 319 g was the dominant fungus responding positively to inoculation with V. dahliae. The results suggest that 1) plant roots select microorganisms from the wider rhizosphere pool, 2) that both rhizosphere soil and root inhabiting fungal communities are influenced by V. dahliae and 3) that soil type has a stronger influence on both of these communities than cultivar. PMID:25347069

  19. Influence of soil type, cultivar and Verticillium dahliae on the structure of the root and rhizosphere soil fungal microbiome of strawberry.

    Science.gov (United States)

    Nallanchakravarthula, Srivathsa; Mahmood, Shahid; Alström, Sadhna; Finlay, Roger D

    2014-01-01

    Sustainable management of crop productivity and health necessitates improved understanding of the ways in which rhizosphere microbial populations interact with each other, with plant roots and their abiotic environment. In this study we examined the effects of different soils and cultivars, and the presence of a soil-borne fungal pathogen, Verticillium dahliae, on the fungal microbiome of the rhizosphere soil and roots of strawberry plants, using high-throughput pyrosequencing. Fungal communities of the roots of two cultivars, Honeoye and Florence, were statistically distinct from those in the rhizosphere soil of the same plants, with little overlap. Roots of plants growing in two contrasting field soils had high relative abundance of Leptodontidium sp. C2 BESC 319 g whereas rhizosphere soil was characterised by high relative abundance of Trichosporon dulcitum or Cryptococcus terreus, depending upon the soil type. Differences between different cultivars were not as clear. Inoculation with the pathogen V. dahliae had a significant influence on community structure, generally decreasing the number of rhizosphere soil- and root-inhabiting fungi. Leptodontidium sp. C2 BESC 319 g was the dominant fungus responding positively to inoculation with V. dahliae. The results suggest that 1) plant roots select microorganisms from the wider rhizosphere pool, 2) that both rhizosphere soil and root inhabiting fungal communities are influenced by V. dahliae and 3) that soil type has a stronger influence on both of these communities than cultivar.

  20. Pattern Analysis of the Interaction between Verticillium dahliae Strains and Cotton Varieties%棉花黄萎病菌与品种互作的格局分析

    Institute of Scientific and Technical Information of China (English)

    吴元奇; 房卫平; 胡秉民

    2009-01-01

    In order to study the interaction pattern of strains and cotton varieties, the disease indices of twenty-three different resistant or susceptible varieties inoculating 18 isolates of Verticillium dahliae were analyzed by biplot method based on cluster analysis. The results showed that 23 cotton cultivars were grouped into six clusters, and 18 Verticillium dahliae strains were clustered into five groups. With the highest resistance to all strain groups, variety cluster G1 which was consisted of two lines (5031 and 50312) could be used as good resistant resource in breeding of Verticillium wilt resistance. There is a high host-pathogen interaction between strain group S4 and variety group G5. As dominant strains and with the highest pathogenicity, S4 consisting of 4 Verticillium dahliae strains among which VD25 came from Chengan of Hebei Province and the rest, named as XS4, XS6, and XS7 all from Changshu of Jiangsu Province, should be very important in identifying and breeding for Verticillium wilt resistance. Strain groups S2 and S3, which highly interacted with 3 variety groups (accounting for 56% of the tested material), might be the potential dominant strains. With average resistance to all strains, variety group G2 was the excellent host for Verticillium dahliae.%以23个棉花品种(系)接种18个黄萎病菌株的抗感表现(病情指数)为分析对象,在聚类分析的基础上用双标图方法研究了寄主-病原的互作格局.结果表明:23个棉花品种分为6个抗病性不同的品种组,18个菌株聚为5个致病力不同的菌株群.5031和50312组成的品种组G1对所有菌株群具有最高抗性,可作为抗黄育种的高抗质源.来自河北成安的VD25与来自江苏常熟的XS4、XS6和XS7组成的菌株群S4与G5品种组存在互作,其致病力最强,是棉花黄萎病的优势菌株群,在抗黄育种和抗黄鉴定中应予以高度重视.S2和S3菌株群与56%的参试品种构成的3个品种组间存在互作,是潜在的

  1. Exploring the Potentials of Lysinibacillus sphaericus ZA9 for Plant Growth Promotion and Biocontrol Activities against Phytopathogenic Fungi

    Directory of Open Access Journals (Sweden)

    Zakira Naureen

    2017-08-01

    Full Text Available There is an ongoing hunt for biologically active compounds that can combat phytopathogenic fungi and improve plant growth without causing any hazards to the environment. Consequently the present study aims at deciphering the plant growth promotion and antifungal capability of Lysinibacillus sphaericus ZA9. The bacterium was previously isolated and identified in our laboratory from maize rhizosphere using 16S rRNA gene sequencing. The test bacterium L. sphaericus ZA9 was found to produce high quantity of IAA (697 μg/ mL; siderophores (195.79 μg/ mL, HCN and hydrolytic enzyme as compared to the reference strain Bacillus sphaericus Z2-7. The bacterium was also capable of solubilizing silicates (Si, phosphates (P, and potassium (K. The bacterium enhanced the seedling vigor and germination of seeds pretreated with it and promoted the shoot length of both cucumber and tomato seeds in greenhouse experiment. L. sphaericus ZA9 and its cell free culture supernatant showed varied antagonistic behavior against Alternaria alternata, Curvularia lunata, Aspergillus sp., Sclerotinia sp., Bipolaris spicifera, Trichophyton sp. Fermentation broth culture of L. sphaericus ZA9 was then used to isolate antifungal metabolites by silica column chromatography. Identification and determination of antifungal compounds was carried out by Thin-layer chromatography (TLC followed by NMR spectroscopy. Two compounds were isolated and identified as 2-pentyl-4-quinolinecarboxylic acid (C15H17NO2 which is a quinoline alkaloid and 1- methylcyclohexene which is a cycloalkene. Compound 1; 2-Penthyl-4-quinolinecarboxylic acid was found to be highly antagonistic against most of the fungi tested as compared to the bacterium itself. Its activity was comparable to that of fungicide Benlate, while compound 2; 1- methylcyclohexene did not show any antifungal activity.

  2. Production, partial purification and characterization of protease from a phytopathogenic fungi Alternaria solani (Ell. and Mart.) Sorauer.

    Science.gov (United States)

    Chandrasekaran, Murugesan; Sathiyabama, Muthukrishnan

    2014-08-01

    An alkaline serine protease producing strain Alternaria solani was optimized for its enzyme production under submerged conditions. The maximum production of protease by A. solani was achieved by using sodium nitrate at the optimum concentration of 0.2% w/v. A. solani produced higher quantities (3.75 [unit/mg of protein]) of an inducible extracellular proteases on day 9 after incubation in czapek's dox broth medium amended with 1% casein as an inducer at pH 8.5, temperature 27 °C and 3% sucrose as carbon source. Extracellular proteases were precipitated by ammonium sulphate saturation (80%) method and purified on Sephadex G-100 column chromatography. The molecular mass of SDS-PAGE and Sephadex G-100 Column Gel permeation chromatography purified protease was estimated to 42 kDa. In addition, trypsin digestion of 42 kDa protein band was carried out and analyzed by MALDI-TOF for the identification of protease. The sequence IKELATNGVVTNVK (378-391) segment of the alkaline serine protease was found by using MS/MS spectrum at 1485 m/z from the purified fraction. It showed optimal activity at 50 °C and pH 9-10 and broad pH stability between pH 6-12. The protease activity was inhibited by phenyl methyl sulfonyl fluoride (PMSF), all the results indicated that the presence of a serine residue in the active site and is thus most likely a member of the serine protease family. This may function as a virulence protein during pathogenesis by A. solani. The results suggested that the presence of appreciable extracellular proteolytic activity in filamentous fungi may serve as a marker of their phytopathogenicity. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. THE IMPACT OF FERTILIZATION AND FOLIAR STIMULATION PRODUCTS BOTH ON INCREASING THE RESISTANCE TO MAJOR PHYTOPATHOGENS ATTACKS, AND ON INCREASING THE QUANTITY AND QUALITY OF WINE GRAPES HARVEST

    Directory of Open Access Journals (Sweden)

    Cristina BUNESCU

    2014-12-01

    Full Text Available The paper aimed to demonstrate the impact of fertilization and foliar stimulation products both on increasing the resistance to major phytopathogens attacks, and on increasing the quantity and quality of wine grapes harvest. Applying the foliar fertilizer products Plonvit Kali (c1, Tytanit (c2 and Optysil (c3 to vines, for a period of three years (2011/2013, in phenophases of intensive growth of shoots and grapes at approved dosages, simultaneously with pesticide treatment, not only a reduction of pathogenic fungi attack was obtained, but also and an increase of harvest without diminishing the quality of the grapes.

  4. Essential oil composition and antifungal activity of Melissa officinalis originating from north-Est Morocco, against postharvest phytopathogenic fungi in apples.

    Science.gov (United States)

    El Ouadi, Y; Manssouri, M; Bouyanzer, A; Majidi, L; Bendaif, H; Elmsellem, H; Shariati, M A; Melhaoui, A; Hammouti, B

    2017-06-01

    To investigate biological control methods against post-harvest phytopathogenic fungi in apples, tests on the antifungal activity of essential oil of Melissa officinalis were carried out. The essential oil, obtained by hydrodistillation, was analyzed by gas chromatography coupled with mass spectrometry (GC-MS). Analysis of the essential oil was able to detect 88.7% of the components. The main components are P-mentha-1,2,3-triol (13.1%), P-menth-3-en-8-ol (8.8%), pulegone (8.8%), piperitynone oxide (8.4%) and 2-piperitone oxide (7.3%). The determination of the antifungal activity of the essential oil of M. officinalisis carried out in vitro using the technique of poison food (PF) and the volatile activity test (VA). To carry out these two tests, three phytopathogens that cause the deterioration of apples have been selected: Botrytis cinerea, Penicillium expansum and Rhizopus stolonifer. The overall results of this study suggest that M. officinalis essential oil has potential as a bio-antifungal preservative for the control of post-harvest diseases of apple. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Analysis of Phaseolus vulgaris response to its association with Trichoderma harzianum (ALL-42) in the presence or absence of the phytopathogenic fungi Rhizoctonia solani and Fusarium solani.

    Science.gov (United States)

    Pereira, Jackeline L; Queiroz, Rayner M L; Charneau, Sébastien O; Felix, Carlos R; Ricart, Carlos A O; da Silva, Francilene Lopes; Steindorff, Andrei Stecca; Ulhoa, Cirano J; Noronha, Eliane F

    2014-01-01

    The present study was carried out to evaluate the ability of Trichoderma harzianum (ALL 42-isolated from Brazilian Cerrado soil) to promote common bean growth and to modulate its metabolism and defense response in the presence or absence of the phytopathogenic fungi Rhizoctonia solani and Fusarium solani using a proteomic approach. T. harzianum was able to promote common bean plants growth as shown by the increase in root/foliar areas and by size in comparison to plants grown in its absence. The interaction was shown to modulate the expression of defense-related genes (Glu1, pod3 and lox1) in roots of P. vulgaris. Proteomic maps constructed using roots and leaves of plants challenged or unchallenged by T. harzianum and phytopathogenic fungi showed differences. Reference gels presented differences in spot distribution (absence/presence) and relative volumes of common spots (up or down-regulation). Differential spots were identified by peptide fingerprinting MALDI-TOF mass spectrometry. A total of 48 identified spots (19 for leaves and 29 for roots) were grouped into protein functional classes. For leaves, 33%, 22% and 11% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively. For roots, 17.2%, 24.1% and 10.3% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively.

  6. A New Synthetic Amphiploid (AADDAA between Gossypium hirsutum and G. arboreum Lays the Foundation for Transferring Resistances to Verticillium and Drought.

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    Yu Chen

    Full Text Available Gossypium arboreum, a cultivated cotton species (2n = 26, AA native to Asia, possesses invaluable characteristics unavailable in the tetraploid cultivated cotton gene pool, such as resistance to pests and diseases and tolerance to abiotic stresses. However, it is quite difficult to transfer favorable traits into Upland cotton through conventional methods due to the cross-incompatibility of G. hirsutum (2n = 52, AADD and G. arboreum. Here, we improved an embryo rescue technique to overcome the cross-incompatibility between these two parents for transferring favorable genes from G. arboreum into G. hirsutum. Our results indicate that MSB2K supplemented with 0.5 mg l(-1 kinetin and 250 mg(-1 casein hydrolysate is an efficient initial medium for rescuing early (3 d after pollination hybrid embryos. Eight putative hybrids were successfully obtained, which were further verified and characterized by cytology, molecular markers and morphological analysis. The putative hybrids were subsequently treated with different concentrations of colchicine solution to double their chromosomes. The results demonstrate that four putative hybrid plants were successfully chromosome-doubled by treatment with 0.1% colchicine for 24 h and become amphiploid, which were confirmed by cytological observation, self-fertilization and backcrossing. Preliminary assessments of resistance at seedling stage indicate that the synthetic amphiploid showed highly resistant to Verticillium and drought. The synthetic amphiploid between G. hirsutum × G. arboreum would lay the foundation for developing G. arboreum-introgressed lines with the uniform genetic background of G. hirsutum acc TM-1, which would greatly enhance and simplify the mining, isolation, characterization, cloning and use of G. arboreum-specific desirable genes in future cotton breeding programs.

  7. A New Synthetic Amphiploid (AADDAA) between Gossypium hirsutum and G. arboreum Lays the Foundation for Transferring Resistances to Verticillium and Drought.

    Science.gov (United States)

    Chen, Yu; Wang, Yingying; Zhao, Ting; Yang, Jianwei; Feng, Shouli; Nazeer, Wajad; Zhang, Tianzhen; Zhou, Baoliang

    2015-01-01

    Gossypium arboreum, a cultivated cotton species (2n = 26, AA) native to Asia, possesses invaluable characteristics unavailable in the tetraploid cultivated cotton gene pool, such as resistance to pests and diseases and tolerance to abiotic stresses. However, it is quite difficult to transfer favorable traits into Upland cotton through conventional methods due to the cross-incompatibility of G. hirsutum (2n = 52, AADD) and G. arboreum. Here, we improved an embryo rescue technique to overcome the cross-incompatibility between these two parents for transferring favorable genes from G. arboreum into G. hirsutum. Our results indicate that MSB2K supplemented with 0.5 mg l(-1) kinetin and 250 mg(-1) casein hydrolysate is an efficient initial medium for rescuing early (3 d after pollination) hybrid embryos. Eight putative hybrids were successfully obtained, which were further verified and characterized by cytology, molecular markers and morphological analysis. The putative hybrids were subsequently treated with different concentrations of colchicine solution to double their chromosomes. The results demonstrate that four putative hybrid plants were successfully chromosome-doubled by treatment with 0.1% colchicine for 24 h and become amphiploid, which were confirmed by cytological observation, self-fertilization and backcrossing. Preliminary assessments of resistance at seedling stage indicate that the synthetic amphiploid showed highly resistant to Verticillium and drought. The synthetic amphiploid between G. hirsutum × G. arboreum would lay the foundation for developing G. arboreum-introgressed lines with the uniform genetic background of G. hirsutum acc TM-1, which would greatly enhance and simplify the mining, isolation, characterization, cloning and use of G. arboreum-specific desirable genes in future cotton breeding programs.

  8. The Impact of Genotyping-by-Sequencing Pipelines on SNP Discovery and Identification of Markers Associated with Verticillium Wilt Resistance in Autotetraploid Alfalfa (Medicago sativa L.)

    Science.gov (United States)

    Yu, Long-Xi; Zheng, Ping; Bhamidimarri, Suresh; Liu, Xiang-Ping; Main, Dorie

    2017-01-01

    Verticillium wilt (VW) of alfalfa is a soilborne disease causing severe yield loss in alfalfa. To identify molecular markers associated with VW resistance, we used an integrated framework of genome-wide association study (GWAS) with high-throughput genotyping by sequencing (GBS) to identify loci associated with VW resistance in an F1 full-sib alfalfa population. Phenotyping was performed using manual inoculation of the pathogen to cloned plants of each individual and disease severity was scored using a standard scale. Genotyping was done by GBS, followed by genotype calling using three bioinformatics pipelines including the TASSEL-GBS pipeline (TASSEL), the Universal Network Enabled Analysis Kit (UNEAK), and the haplotype-based FreeBayes pipeline (FreeBayes). The resulting numbers of SNPs, marker density, minor allele frequency (MAF) and heterozygosity were compared among the pipelines. The TASSEL pipeline generated more markers with the highest density and MAF, whereas the highest heterozygosity was obtained by the UNEAK pipeline. The FreeBayes pipeline generated tetraploid genotypes, with the least number of markers. SNP markers generated from each pipeline were used independently for marker-trait association. Markers significantly associated with VW resistance identified by each pipeline were compared. Similar marker loci were found on chromosomes 5, 6, and 7, whereas different loci on chromosome 1, 2, 3, and 4 were identified by different pipelines. Most significant markers were located on chromosome 6 and they were identified by all three pipelines. Of those identified, several loci were linked to known genes whose functions are involved in the plants’ resistance to pathogens. Further investigation on these loci and their linked genes would provide insight into understanding molecular mechanisms of VW resistance in alfalfa. Functional markers closely linked to the resistance loci would be useful for MAS to improve alfalfa cultivars with enhanced resistance

  9. Genome-wide identification of HrpL-regulated genes in the necrotrophic phytopathogen Dickeya dadantii 3937.

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    Shihui Yang

    Full Text Available BACKGROUND: Dickeya dadantii is a necrotrophic pathogen causing disease in many plants. Previous studies have demonstrated that the type III secretion system (T3SS of D. dadantii is required for full virulence. HrpL is an alternative sigma factor that binds to the hrp box promoter sequence of T3SS genes to up-regulate their expression. METHODOLOGY/PRINCIPAL FINDINGS: To explore the inventory of HrpL-regulated genes of D. dadantii 3937 (3937, transcriptome profiles of wild-type 3937 and a hrpL mutant grown in a T3SS-inducing medium were examined. Using a cut-off value of 1.5, significant differential expression was observed in sixty-three genes, which are involved in various cellular functions such as type III secretion, chemotaxis, metabolism, regulation, and stress response. A hidden Markov model (HMM was used to predict candidate hrp box binding sites in the intergenic regions of 3937, including the promoter regions of HrpL-regulated genes identified in the microarray assay. In contrast to biotrophic phytopathgens such as Pseudomonas syringae, among the HrpL up-regulated genes in 3937 only those within the T3SS were found to contain a hrp box sequence. Moreover, direct binding of purified HrpL protein to the hrp box was demonstrated for hrp box-containing DNA fragments of hrpA and hrpN using the electrophoretic mobility shift assay (EMSA. In this study, a putative T3SS effector DspA/E was also identified as a HrpL-upregulated gene, and shown to be translocated into plant cells in a T3SS-dependent manner. CONCLUSION/SIGNIFICANCES: We provide the genome-wide study of HrpL-regulated genes in a necrotrophic phytopathogen (D. dadantii 3937 through a combination of transcriptomics and bioinformatics, which led to identification of several effectors. Our study indicates the extent of differences for T3SS effector protein inventory requirements between necrotrophic and biotrophic pathogens, and may allow the development of different strategies for

  10. Serine protease identification (in vitro) and molecular structure predictions (in silico) from a phytopathogenic fungus, Alternaria solani.

    Science.gov (United States)

    Chandrasekaran, Murugesan; Chandrasekar, Raman; Sa, Tongmin; Sathiyabama, Muthukrishnan

    2014-07-01

    Serine proteases are involved in an enormous number of biological processes. The present study aims at characterizing three-dimensional (3D) molecular architecture of serine proteases from early blight pathogen, Alternaria solani that are hypothesized to be markers of phytopathogenicity. A serine protease was purified to homogeneity and MALDI-TOF-MS/MS analysis revealed that protease produced by A. solani belongs to alkaline serine proteases (AsP). AsP is made up of 403 amino acid residues with molecular weight of 42.1 kDa (Isoelectric point - 6.51) and its molecular formula was C1859 H2930 N516 O595 S4 . AsP structure model was built based on its comparative homology with serine protease using the program, MODELER. AsP had 16 β-sheets and 10 α-helices, with Ser(350) (G347-G357), Asp(158) (D158-H169), and His(193) (H193-G203) in separate turn/coil structures. Biological metal binding region situated near 6th-helix and His(193) residue is responsible for metal binding site. Also, calcium ion (Ca(2+)) is coordinated by the carboxyl groups of Lys(84), Ile(85), Lys(86), Asp(87), Phe(88), Ala(89), Ala(90) (K84-A90) for first Ca(2+) binding site and carbonyl oxygen atom of Lys(244), Gly(245), Arg(246), Thr(247), Lys(248), Lys(249), and Ala(250) (K244-A250), for second Ca(2+) binding site. Moreover, Ramachandran plot analysis of protein residues falling into most favored secondary structures were determined (83.3%). The predicted molecular 3D structural model was further verified using PROCHECK, ERRAT, and VADAR servers to confirm the geometry and stereo-chemical parameters of the molecular structural design. The functional analysis of AsP 3D molecular structure predictions familiar in the current study may provide a new perspective in the understanding and identification of antifungal protease inhibitor designing. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Anti-phytopathogen potential of endophytic actinobacteria isolated from tomato plants (Lycopersicon esculentum) in southern Brazil, and characterization of Streptomyces sp. R18(6), a potential biocontrol agent.

    Science.gov (United States)

    de Oliveira, Margaroni Fialho; da Silva, Mariana Germano; Van Der Sand, Sueli T

    2010-09-01

    Tomato plants (Lycopersicon esculentum) are highly susceptible to phytopathogen attack. The resulting intensive application of pesticides on tomato crops can affect the environment and health of humans and animals. The objective of this study was to select potential biocontrol agents among actinobacteria from tomato plants, in a search for alternative phytopathogen control. We evaluated 70 endophytic actinobacteria isolated from tomato plants in southern Brazil, testing their antimicrobial activity, siderophore production, indoleacetic acid production, and phosphate solubility. The actinomycete isolate with the highest antimicrobial potential was selected using the agar-well diffusion method, in order to optimize conditions for the production of compounds with antimicrobial activity. For this study, six growth media (starch casein-SC, ISP2, Bennett's, Sahin, Czapek-Dox, and TSB), three temperatures (25 degrees C, 30 degrees C, and 35 degrees C) and different pH were tested. Of the actinobacteria tested, 88.6% showed antimicrobial activity against at least one phytopathogen, 72.1% showed a positive reaction for indoleacetic acid production, 86.8% produced siderophores and 16.2% showed a positive reaction for phosphate solubility. Isolate R18(6) was selected due to its antagonistic activity against all phytopathogenic microorganisms tested in this study. The best conditions for production were observed in the SC medium, at 30 degrees C and pH 7.0. The isolate R18(6) showed close biochemical and genetic similarity to Streptomyces pluricolorescens.

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

    Science.gov (United States)

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

    2015-01-29

    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.

  13. Molecular Variability Within and Among Verticillium dahliae Vegetative Compatibility Groups Determined by Fluorescent Amplified Fragment Length Polymorphism and Polymerase Chain Reaction Markers.

    Science.gov (United States)

    Collado-Romero, Melania; Mercado-Blanco, Jesús; Olivares-García, Concepción; Valverde-Corredor, Antonio; Jiménez-Díaz, Rafael M

    2006-05-01

    ABSTRACT A degree of genetic diversity may exist among Verticillium dahliae isolates within vegetative compatibility groups (VCGs) that bears phytopathological significance and is worth investigating using molecular tools of a higher resolution than VCG characterization. The molecular variability within and among V. dahliae VCGs was studied using 53 artichoke isolates from eastern-central Spain, 96 isolates from cotton, 7 from cotton soil, and 45 from olive trees in countries of the Mediterranean Basin. Isolates were selected to represent the widest available diversity in cotton- and olive-defoliating (D) and -nondefoliating (ND) pathotypes, as well as for VCG. The VCG of 96 cotton and olive isolates was determined in this present study. Molecular variability among V. dahliae isolates was assessed by fluorescent amplified fragment length polymorphism (AFLP) analysis and by polymerase chain reaction (PCR) assays for DNA fragments associated with the D (462 bp) and ND (824 bp) pathotypes, as well as a 334-bp amplicon associated with D pathotype isolates but also present in some VCG2B isolates. Isolates from cotton were in VCG1A, VCG1B, VCG2A, VCG2B, and VCG4B and those from olive trees were in VCG1A, VCG2A, and VCG4B. Artichoke isolates included representatives of VCG1A, VCG2A, VCG2B (including a newly identified VCG2Ba), and VCG4B. AFLP data were used to generate matrixes of genetic distance among isolates for cluster analysis using the neighbor-joining method and for analysis of molecular variance. Results demonstrated that V. dahliae isolates within a VCG subgroup are molecularly similar, to the extent that clustering of isolates correlated with VCG subgroups regardless of the host source and geographic origin. VCGs differed in molecular variability, with the variability being highest in VCG2B and VCG2A. For some AFLP/VCG subgroup clusterings, V. dahliae isolates from artichoke grouped in subclusters clearly distinct from those comprising isolates from cotton and

  14. Rhizobium rubi(T): a gram-negative phytopathogenic bacterium expressing the Lewis B epitope on the outer core of its lipooligosaccharide fraction.

    Science.gov (United States)

    Gargiulo, Valentina; Garozzo, Domenico; Lanzetta, Rosa; Molinaro, Antonio; Sturiale, Luisa; De Castro, Cristina; Parrilli, Michelangelo

    2008-07-21

    The structure of the core oligosaccharide from the phytopathogenic bacterium Rhizobium rubi was deduced by combining information from complementary chemical approaches (alkaline and acid hydrolysis), similar to the "overlap peptide" strategy. This structure is new and it contains two main oligosaccharide backbones that differ in the substitution degree of the external Kdo unit. The relevant feature shared by both oligosaccharides is the presence of a tetrasaccharide motif that is similar to the blood group Lewis B antigen (Le(B)). This epitope differs from Le(B) in the glycosidic configuration of the glucosamine unit (alpha and not beta) and in the occurrence of acetyls substituents at O3 and/or O4 of the galactose moiety. Other notable structural features are the location of the Dha residue, the presence of a alpha-glucose unit that is linked to the inner Kdo unit, the high number of acid sugars and the highly branched core structure.

  15. Structural and functional characterization of the GalNAc/Gal-specific lectin from the phytopathogenic ascomycete Sclerotinia sclerotiorum (Lib.) de Bary.

    Science.gov (United States)

    Candy, Laure; Van Damme, Els J M; Peumans, Willy J; Menu-Bouaouiche, Laurence; Erard, Monique; Rougé, Pierre

    2003-08-22

    The lectin found in mycelium and sclerotes of the phytopathogenic fungus Sclerotinia sclerotiorum is a homodimer consisting of two identical non-covalently bound subunits of 16,000 Da. CD spectra analysis revealed that the S. sclerotiorum agglutinin (SSA) contains predominantly beta-sheet structures. SSA exhibits specificity towards GalNAc whereby the hydroxyls at positions 4 and 6 of the pyranose ring play a key role in the interaction with simple sugars. The carbohydrate-binding site of SSA can also accommodate disaccharides. The N-terminal sequence of SSA shares no significant similarity with any other protein except a lectin from the Sclerotiniaceae species Ciborinia camelliae. A comparison of SSA and the lectins from C. camelliae and some previously characterized lectins indicates that the Sclerotiniaceae lectins form a homogeneous family of fungal lectins. This newly identified lectin family, which is structurally unrelated to any other family of fungal lectins, is most probably confined to the Ascomycota.

  16. Heterologous expression of the glucose oxidase gene in Trichoderma atroviride leads enhanced ability to attack phytopathogenic fungi and induction of plant systemic disease resistance

    Institute of Scientific and Technical Information of China (English)

    Robert L Mach; Brunner Kurt; Matteo Lorito; Susanne Zeilinger; Rosalia Ciliento; Sheridan Woo

    2004-01-01

    @@ A transgenic strain of Trichoderma atroviride that expresses the Aspergillus niger glucose oxidase gene goxA under a homologous pathogen-inducible promoter (nag1) has been constructed, with the aim of increasing the ability of this biocontrol agent (BCA) to attack phytopathogenic fungi and enhance plant systemic disease resistance. The sporulation and growth rate of the transgenic progenies were similar to the wild-type strain Pl. goxA expression occurred immediately after contact with the plant pathogen,and the glucose oxidase formed was secreted extracellularly. The transformed strain SJ3 4, containing 12-14 copies of the transgene, produced significantly less N-acetyl-glucosaminidase and endochitinase then wild type. However, the ability of its culture filtrate to inhibit the germination of Botrytis cinerea spores was increased by about 3-fold. In comparison to P1, the transgenic strain more quickly overgrew and lysed in vitro the pathogens Rhizoctonia solani and Pythium ultimum.

  17. The alternative sigma factor HrpL negatively modulates the flagellar system in the phytopathogenic bacterium Erwinia amylovora under hrp-inducing conditions.

    Science.gov (United States)

    Cesbron, Sophie; Paulin, Jean-Pierre; Tharaud, Michel; Barny, Marie-Anne; Brisset, Marie-Noëlle

    2006-04-01

    In this work we present evidence of an opposite regulation in the phytopathogenic bacteria Erwinia amylovora between the virulence-associated Type III secretion system (TTSS) and the flagellar system. Using loss-of-function mutants we show that motility enhanced the virulence of wild-type bacteria relative to a nonmotile mutant when sprayed on apple seedlings with unwounded leaves. Then we demonstrated through analyses of motility, flagellin export and visualization of flagellar filament that HrpL, the positive key regulator of the TTSS, also down-regulates the flagellar system. Such a dual regulation mediated by an alternative sigma factor of the TTSS appears to be a level of regulation between virulence and motility not yet described among Proteobacteria.

  18. Screening and identification of intagonistic bacterium strain 8-52 against Verticillium dahliae%大丽轮枝菌拮抗细菌8-52菌株的筛选与鉴定

    Institute of Scientific and Technical Information of China (English)

    吴国江; 王占利; 李术娜; 王全; 李红亚; 朱宝成

    2009-01-01

    为筛选对棉花黄萎病病原菌大丽轮枝菌(Verticillium dahliae)具有拮抗作用的芽孢细菌,采用改良的琼脂平板扩散法,通过初筛从各地的土样中分离到83株拮抗细菌菌株,对其中的69株进行了复筛,得到一株具有较强的抑菌活性的拮抗细菌8-52菌株.对该菌株进行了形态特征的观察、生理生化试验和16S rDNA的序列分析.将该菌株的形态特征和生理生化特性鉴定的结果与相应种、属模式菌的性状相对照,认为菌株8-52与芽孢杆菌(Bacillus)的相应性状很相近.根据16S rDNA序列相似性分析,此菌株与Bacillus velezensis标准菌株AB245422的16S rDNA序列相似度达99.78%,因此鉴定拮抗菌株8-52为Bacillus velezensis.%In order to obtain antagonistic bacteria against Verticillium dahliae, 83 antagonistic strains were isolated and screened with the improved agar plate diffusion method from the soil of different districts. 69 strains of them were tested their antagonistic activity against Verticillium dahliae Kleb via secondary screening. As a result, a strain called 8-52 which had strong inhibition capability against Verticillum dahliae strain V-190 was obtained. Then the strain 8-52 was characterized by morphological and culture features observation, physiological and biochemical experiments, and 16S rDNA sequence analysis. As the morphology characteristics and physiological and biochemical characteristics of the stain are similar to Bacillus and the similarity of the 16S rDNA sequences between strain 8-52 and the type strain was up to 99.21%, the strain 8-52 was initially identified as Bacillus malacitensis.

  19. lpxC and yafS are the most suitable internal controls to normalize real time RT-qPCR expression in the phytopathogenic bacteria Dickeya dadantii.

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    Florence Hommais

    Full Text Available BACKGROUND: Quantitative RT-PCR is the method of choice for studying, with both sensitivity and accuracy, the expression of genes. A reliable normalization of the data, using several reference genes, is critical for an accurate quantification of gene expression. Here, we propose a set of reference genes, of the phytopathogenic bacteria Dickeya dadantii and Pectobacterium atrosepticum, which are stable in a wide range of growth conditions. RESULTS: We extracted, from a D. dadantii micro-array transcript profile dataset comprising thirty-two different growth conditions, an initial set of 49 expressed genes with very low variation in gene expression. Out of these, we retained 10 genes representing different functional categories, different levels of expression (low, medium, and high and with no systematic variation in expression correlating with growth conditions. We measured the expression of these reference gene candidates using quantitative RT-PCR in 50 different experimental conditions, mimicking the environment encountered by the bacteria in their host and directly during the infection process in planta. The two most stable genes (ABF-0017965 (lpxC and ABF-0020529 (yafS were successfully used for normalization of RT-qPCR data. Finally, we demonstrated that the ortholog of lpxC and yafS in Pectobacterium atrosepticum also showed stable expression in diverse growth conditions. CONCLUSIONS: We have identified at least two genes, lpxC (ABF-0017965 and yafS (ABF-0020509, whose expressions are stable in a wide range of growth conditions and during infection. Thus, these genes are considered suitable for use as reference genes for the normalization of real-time RT-qPCR data of the two main pectinolytic phytopathogenic bacteria D. dadantii and P. atrosepticum and, probably, of other Enterobacteriaceae. Moreover, we defined general criteria to select good reference genes in bacteria.

  20. Effect of crude plant extracts from some Oaxacan flora on two deleterious fungal phytopathogens and extract compatibility with a biofertilizer strain

    Directory of Open Access Journals (Sweden)

    Karla Isabel eLira-De León

    2014-08-01

    Full Text Available The antimicrobial activity of twelve plant extracts was tested against the phytopathogens Alternaria alternata and Fusarium solani. In addition, the compatibility of the extracts towards Bacillus liqueniformis, a biofertilizer and a non-target microorganism, was assessed. Plants tested belong to Euphorbiaceae, Asteraceae, Crassulaceae, Rubiaceae, Convolvulaceae, Verbenaceae, Orchidaceae, Nyctaginaceae, Boraginaceae, and Tiliaecae families and were collected in the State of Oaxaca. The antifungal activity of the plant extracts (50 to 100 mg/mL against A. alternata and F. solani, was determined by measuring the mycelium radial growth and obtaining the minimum inhibitory concentration (MIC of fungal growth. In addition, with the aim of finding plant extracts which are compatible with a B. licheniformis biofertilizer strain and to test the nontoxic nature of the treatments, the toxicity of the extracts towards this strain was evaluated using the agar diffusion method. Azoxystrobin (12 μg and chloramphenicol (30 μg were used as positive controls for the pathogens and for the non-target bacteria, respectively. Plant extracts inhibited fungal growth in the ranges of 0.76 to 56.17 % against F. solani and 2.02 to 69.07 % against A. alternata. The extracts of Acalypha subviscida, Ipomoea murucoides, Tournefortia densiflora and Lantana achyranthifolia showed MIC values between 5.77-12.5 mg/mL for at least one of the fungal species. The best treatment, Adenophyllum aurantium, exhibited a maximum inhibition for both F. solani (56.17%, MIC = 7.78 mg/mL and A. alternata (68.64% MIC = 7.78 mg/mL, and resulted innocuous toward B. licheniformis. Therefore, this plant has an outstanding potential for the agroecological control of fungal phytopathogens in industrial crops.

  1. Effect of crude plant extracts from some Oaxacan flora on two deleterious fungal phytopathogens and extract compatibility with a biofertilizer strain

    Science.gov (United States)

    Lira-De León, Karla I.; Ramírez-Mares, Marco V.; Sánchez-López, Vladimir; Ramírez-Lepe, Mario; Salas-Coronado, Raúl; Santos-Sánchez, Norma F.; Valadez-Blanco, Rogelio; Hernández-Carlos, Beatriz

    2014-01-01

    The antimicrobial activity of 12 plant extracts was tested against the phytopathogens Alternaria alternata and Fusarium solani. In addition, the compatibility of the extracts toward Bacillus liqueniformis, a biofertilizer and a non-target microorganism, was assessed. Plants tested belong to the Euphorbiaceae, Asteraceae, Crassulaceae, Rubiaceae, Convolvulaceae, Verbenaceae, Orchidaceae, Nyctaginaceae, Boraginaceae, and Tiliaceae families and were collected in the State of Oaxaca. The antifungal activity of the plant extracts (50–100 mg/mL) against A. alternata and F. solani, was determined by measuring the mycelium radial growth and obtaining the minimum inhibitory concentration (MIC) of fungal growth. In addition, with the aim of finding plant extracts which are compatible with a B. licheniformis biofertilizer strain and to test the non-toxic nature of the treatments, the toxicity of the extracts toward this strain was evaluated using the agar diffusion method. Azoxystrobin (12 μg) and chloramphenicol (30 μg) were used as positive controls for the pathogens and for the non-target bacteria, respectively. Plant extracts inhibited fungal growth in the ranges of 0.76–56.17% against F. solani and 2.02–69.07% against A. alternata. The extracts of Acalypha subviscida, Ipomoea murucoides, Tournefortia densiflora and Lantana achyranthifolia showed MIC values between 5.77–12.5 mg/mL for at least one of the fungal species. The best treatment, Adenophyllum aurantium, exhibited a maximum inhibition for both F. solani (56.17%, MIC = 7.78 mg/mL) and A. alternata (68.64% MIC = 7.78 mg/mL), and resulted innocuous toward B. licheniformis. Therefore, this plant has an outstanding potential for the agroecological control of fungal phytopathogens in industrial crops. PMID:25147544

  2. Effect of crude plant extracts from some Oaxacan flora on two deleterious fungal phytopathogens and extract compatibility with a biofertilizer strain.

    Science.gov (United States)

    Lira-De León, Karla I; Ramírez-Mares, Marco V; Sánchez-López, Vladimir; Ramírez-Lepe, Mario; Salas-Coronado, Raúl; Santos-Sánchez, Norma F; Valadez-Blanco, Rogelio; Hernández-Carlos, Beatriz

    2014-01-01

    The antimicrobial activity of 12 plant extracts was tested against the phytopathogens Alternaria alternata and Fusarium solani. In addition, the compatibility of the extracts toward Bacillus liqueniformis, a biofertilizer and a non-target microorganism, was assessed. Plants tested belong to the Euphorbiaceae, Asteraceae, Crassulaceae, Rubiaceae, Convolvulaceae, Verbenaceae, Orchidaceae, Nyctaginaceae, Boraginaceae, and Tiliaceae families and were collected in the State of Oaxaca. The antifungal activity of the plant extracts (50-100 mg/mL) against A. alternata and F. solani, was determined by measuring the mycelium radial growth and obtaining the minimum inhibitory concentration (MIC) of fungal growth. In addition, with the aim of finding plant extracts which are compatible with a B. licheniformis biofertilizer strain and to test the non-toxic nature of the treatments, the toxicity of the extracts toward this strain was evaluated using the agar diffusion method. Azoxystrobin (12 μg) and chloramphenicol (30 μg) were used as positive controls for the pathogens and for the non-target bacteria, respectively. Plant extracts inhibited fungal growth in the ranges of 0.76-56.17% against F. solani and 2.02-69.07% against A. alternata. The extracts of Acalypha subviscida, Ipomoea murucoides, Tournefortia densiflora and Lantana achyranthifolia showed MIC values between 5.77-12.5 mg/mL for at least one of the fungal species. The best treatment, Adenophyllum aurantium, exhibited a maximum inhibition for both F. solani (56.17%, MIC = 7.78 mg/mL) and A. alternata (68.64% MIC = 7.78 mg/mL), and resulted innocuous toward B. licheniformis. Therefore, this plant has an outstanding potential for the agroecological control of fungal phytopathogens in industrial crops.

  3. Effects of different fertilization treatments on soil fertility, verticillium wilt and yield of eggplant%不同肥料对土壤肥力、茄子黄萎病和产量的影响

    Institute of Scientific and Technical Information of China (English)

    帅正彬; 席春奕; 杨斌; 郭江洪; 赵开平

    2016-01-01

    Using ‘Huaxiajiaozi’ eggplant as the material, a field experiment was conducted to evaluate the effects of different fertilization treatments on soil fertility, microorganism’s community structure, and resistance to verticillium wilt. Compared to the control, treatments adding microbial compound fertilizer or biological organic fertilizer could improve the nutrient structure of soil, optimize the structure of microbial flora, reduce the incidence of eggplant verticillium wilt, and increase the yield. Considering the factors of soil fertility, resistance to disease and yield of eggplant, treatment of 22 kg/667 m2 calcium magnesium phosphate, 8 kg/667 m2 potassium chloride, 100 kg/667 m2 shanxichangxin microbial compound fertilizer, was recommended as the optimal fertilizer combination in planting eggplant.%以‘华夏骄子’茄为试验材料,研究不同施肥处理对土壤营养状况、微生物特性、茄子抗黄萎病和产量的影响。结果表明,与常规施肥相比,其他施肥处理可不同程度地提高土壤养分、优化微生物菌群结构,同时能降低茄子黄萎病的发病率、提高茄子产量。综合考虑土壤、抗病性和产量等因素,推荐最佳施肥方案为钙镁磷肥22 kg/667 m2+氯化钾8 kg/667 m2+山西昌鑫微生物复合肥100 kg/667 m2。

  4. Fontes de resistência em acessos de Solanum (secção Lycopersicon a Verticillium dahliae raças 1 e 2 Sources of resistance to Verticillium dahliae races 1 and 2 in accessions of Solanum (section Lycopersicon

    Directory of Open Access Journals (Sweden)

    Bruno Eduardo C de Miranda

    2010-12-01

    Full Text Available A murcha-de-verticílio (Verticillium dahliae é uma das doenças mais severas do tomateiro (Solanum lycopersicum L.. Duas raças fisiológicas de V. dahliae foram caracterizadas infectando o tomateiro e ambas estão presentes no Brasil. A estratégia de controle mais efetiva para isolados da raça 1 tem sido a utilização de cultivares resistentes em associação com práticas culturais. Entretanto, não existem cultivares comerciais de tomate resistentes à raça 2 do patógeno. Neste contexto, torna-se importante a identificação de novas fontes de resistência que sejam efetivas contra as duas raças de V. dahliae. Buscou-se fontes de resistência a isolados de V. dahliae (raças 1 e 2. Cem acessos de uma coleção de germoplasma contendo espécies cultivadas e selvagens de Solanum (secção Lycopersicon foram inicialmente inoculados pelo método de imersão de raízes com um isolado de V. dahliae raça 1 (5 mL de uma suspensão de 10(6 conídios/mL em condições de casa de vegetação. A avaliação foi feita aos 30 dias após a inoculação, usando uma escala de notas variando de 1 (planta sadia a 5 (planta morta. Foram considerados resistentes os acessos que apresentaram nota média entre 1 e 2. Um grupo composto por 38 acessos classificados como resistentes neste primeiro ensaio foi reavaliado para resistência a outros quatro isolados de V. dahliae pertencentes às raças 1 e 2. As cultivares Ponderosa e Floradade foram utilizadas como controles. O ensaio foi conduzido em casa de vegetação em delineamento experimental de blocos ao acaso em arranjo fatorial 5 x 40, com três repetições (vasos com quatro plantas. A avaliação foi feita baseada nos parâmetros epidemiológicos: período de incubação e índice de doença. Foram identificados acessos com resistência raça-específica e também com resistência a ambas as raças. Estes acessos podem ser indicados para futuros programas de melhoramento genético visando incorporar

  5. Busca de fontes de resistência genética à verticillium dahliae em solanum melongena, s. stramonifolium e s. scuticum e manejo da irrigação para controle da murcha de verticílio em berinjela

    OpenAIRE

    Cabral, Ricardo Nunes

    2016-01-01

    A berinjela (Solanum melongena L.) éuma das hortaliças mais rústicas , porém,é suscetível a algumas doenças, principalmente fúngicas. A murcha de verticilio (Verticillium dahliae) é uma doença de manejo difícil, pois o controle curativo é impossível na prática e o preventivo nem sempre é eficaz, devido à capacidade de sobrevivência do patógeno no solo. Não há cultivares comerciais de berinjela resistentes ao Verticillium, o que torna atualmente inviável o controle por resistência genética. O ...

  6. 向日葵黄萎病拮抗细菌Y1-2菌株的鉴定与抑制效果%IDENTIFICATION AND INHIBITION EFFECT OF VERTICILLIUM WILT OF SUNFLOWER ANTAGONISM BACTERIA STRAIN Y1 -2

    Institute of Scientific and Technical Information of China (English)

    王颖; 周臣; 张一名; 赵君; 周洪友

    2011-01-01

    从向日葵黄萎病病区的健康植株根际分离到l株对向日葵黄萎病病原菌(Verticillium dahliae Kleb.)具有较强拮抗作用的菌株Y1-2.经形态观察、生理生化测定及16S rDNA序列对比分析结果表明,菌株Y1-2鉴定为Bacillus subtilis,其与(CJT14XVY01S)B.subtilis序列相似性达99.96%.菌株Y1 -2与大丽轮枝菌对峙培养可以形成4 mm的抑菌带,室内防治试验表明Y1 -2菌株对向日葵黄萎病的防治效果为75.3%;在田间小区防治试验中,Y1-2菌株对向日葵黄萎病的防治效果为81.8%.%We isolated the Verticillium wilt of sunflower pathogen( Verticillium dahliae Kleb. ) antagonist bacterial stain Yl -2 from the soil of sunflower roots. The identification results via morphology, physiological and biochemical characters and 16S rDNA analysis suggested that Yl -2 has 99.96% similarity with (CJT14XVY01S) B. Subtilis. The mutual confrontation training of stain Yl -2 and Verticillium dahliae Kleb. Results showed the width of the inhibition belt is 4 mm. The greenhouse control effect of Yl -2 is 75. 3% , and the field control effect reaches to 81. 8 %.

  7. First interception of Verticillium albo-atrum in Fangchenggang of Guangxi%广西防城港首次截获苜蓿黄萎病菌

    Institute of Scientific and Technical Information of China (English)

    黄胜光; 卢兆山; 邱世明; 奚国华; 陆道; 钟恒禄

    2012-01-01

    从一批进境的美国苜蓿草样品中保湿分离得到了3株与苜蓿黄萎病菌(Verticillium albo-atrum)相似的目标菌VA1、VA2、VA3.该目标菌在PDA上生长迅速,菌丝生长0.8~1.0 mm/d,2~3 d即可生成肉眼可见的菌落斑点;菌落为白色圆形,边缘规则呈圆形,菌丝较致密,气生菌丝较少,菌落中心乳白色奶冻状;培养4~5 d即后产生气生典型轮枝状分生孢子梗,7~8 d后菌落中央表面因产生休眠菌丝开始变成黑褐色至黑色,20 d后菌落的表面和背面大部分均变黑色,仍不产生微菌核和厚垣孢子.该目标菌的纯培养用V.albo-atrum特异引物Vaa1/Vaa2进行检测,PCR 扩增后得到预期330 bp的产物片段,产物序列与V.albo-atrum相应序列的相似性为100%.从而鉴定确认该目标菌为苜蓿黄萎病菌V.albo-atrum.%Three strains of the fungus, namely VA1, VA2 and VA3, were isolated from a batch of alfalfa samples, which were imported from USA and incubated under moist conditions, demonstrating the characters similar to V. Albo-atrum. The strains grew rapidly on the PDA, with the colony growing at 0.8 - 1. Omm/d. After 2-3 days, the spot colony could be seen on the PDA. In addition, the colony had a regular and rounded margin with white compact mycelium and fewer aerial mycelia. The target fungus produced typical round aerial branched co-nidiophores after 4-5 days, and then became dark brown to black along with production of resting hyphae at the centre of the colony after 7-8 days, and the color of the surface and most of the back of the colony changed to black 20 days later, but still could not produce micro-sclerotia and chlamydospores. The pure cultures of the target germ were detected by the specific primers Vaal/Vaa2. As expected, a 330 bp fragment was obtained by PCR amplification. The similarity between the product and corresponding sequence in V. Albo-atrum was 100%. Therefore, it was confirmed that the target germ was V. Albo-atrum.

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

    Science.gov (United States)

    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

  9. Verticillium wilt of Fraxinus excelsior

    NARCIS (Netherlands)

    Hiemstra, J.A.

    1995-01-01

    Research on ash wilt disease, a common disease of Fraxinus excelsior L. in young forest and landscape plantings in several parts of the Netherlands, is described. By means of a survey for pathogenic fungi in affected trees, inoculation and

  10. The Transition from a Phytopathogenic Smut Ancestor to an Anamorphic Biocontrol Agent Deciphered by Comparative Whole-Genome Analysis[W][OPEN

    Science.gov (United States)

    Lefebvre, François; Joly, David L.; Labbé, Caroline; Teichmann, Beate; Linning, Rob; Belzile, François; Bakkeren, Guus; Bélanger, Richard R.

    2013-01-01

    Pseudozyma flocculosa is related to the model plant pathogen Ustilago maydis yet is not a phytopathogen but rather a biocontrol agent of powdery mildews; this relationship makes it unique for the study of the evolution of plant pathogenicity factors. The P. flocculosa genome of ∼23 Mb includes 6877 predicted protein coding genes. Genome features, including hallmarks of pathogenicity, are very similar in P. flocculosa and U. maydis, Sporisorium reilianum, and Ustilago hordei. Furthermore, P. flocculosa, a strict anamorph, revealed conserved and seemingly intact mating-type and meiosis loci typical of Ustilaginales. By contrast, we observed the loss of a specific subset of candidate secreted effector proteins reported to influence virulence in U. maydis as the singular divergence that could explain its nonpathogenic nature. These results suggest that P. flocculosa could have once been a virulent smut fungus that lost the specific effectors necessary for host compatibility. Interestingly, the biocontrol agent appears to have acquired genes encoding secreted proteins not found in the compared Ustilaginales, including necrosis-inducing-Phytophthora-protein- and Lysin-motif- containing proteins believed to have direct relevance to its lifestyle. The genome sequence should contribute to new insights into the subtle genetic differences that can lead to drastic changes in fungal pathogen lifestyles. PMID:23800965

  11. FTIR spectroscopic evaluation of changes in the cellular biochemical composition of the phytopathogenic fungus Alternaria alternata induced by extracts of some Greek medicinal and aromatic plants

    Science.gov (United States)

    Skotti, Efstathia; Kountouri, Sophia; Bouchagier, Pavlos; Tsitsigiannis, Dimitrios I.; Polissiou, Moschos; Tarantilis, Petros A.

    2014-06-01

    In this study, the biological activity of aquatic extracts of selected Greek medicinal and aromatic plants to the phytopathogenic fungus Alternaria alternata was investigated. Lamiaceae species (Hyssopus officinalis L., Melissa officinalis L., Origanum dictamnus L., Origanum vulgare L. and Salvia officinalis L.) were found to enhance significantly the mycelium growth whereas Crocus sativus appears to inhibit it slightly. M. officinalis and S. officinalis caused the highest stimulation in mycelium growth (+97%) and conidia production (+65%) respectively. In order to further investigate the bioactivity of plant extracts to A. alternata, we employed Fourier Transform Infrared Spectroscopy (FTIR). Differences of original spectra were assigned mainly to amides of proteins. The second derivative transformation of spectra revealed changes in spectral regions corresponding to absorptions of the major cellular constituents such as cell membrane and proteins. Principal component analysis of the second derivative transformed spectra confirmed that fatty acids of the cell membranes, amides of proteins and polysaccharides of the cell wall had the major contribution to data variation. FTIR band area ratios were found to correlate with fungal mycelium growth.

  12. Green synthesis of protein capped silver nanoparticles from phytopathogenic fungus Macrophomina phaseolina (Tassi) Goid with antimicrobial properties against multidrug-resistant bacteria

    Science.gov (United States)

    Chowdhury, Supriyo; Basu, Arpita; Kundu, Surekha

    2014-07-01

    In recent years, green synthesis of nanoparticles, i.e., synthesizing nanoparticles using biological sources like bacteria, algae, fungus, or plant extracts have attracted much attention due to its environment-friendly and economic aspects. The present study demonstrates an eco-friendly and low-cost method of biosynthesis of silver nanoparticles using cell-free filtrate of phytopathogenic fungus Macrophomina phaseolina. UV-visible spectrum showed a peak at 450 nm corresponding to the plasmon absorbance of silver nanoparticles. Scanning electron microscopy (SEM), atomic force microscopy (AFM), and transmission electron microscopy (TEM) revealed the presence of spherical silver nanoparticles of the size range 5 to 40 nm, most of these being 16 to 20 nm in diameter. X-ray diffraction (XRD) spectrum of the nanoparticles exhibited 2 θ values corresponding to silver nanoparticles. These nanoparticles were found to be naturally protein coated. SDS-PAGE analysis showed the presence of an 85-kDa protein band responsible for capping and stabilization of the silver nanoparticles. Antimicrobial activities of the silver nanoparticles against human as well as plant pathogenic multidrug-resistant bacteria were assayed. The particles showed inhibitory effect on the growth kinetics of human and plant bacteria. Furthermore, the genotoxic potential of the silver nanoparticles with increasing concentrations was evaluated by DNA fragmentation studies using plasmid DNA.

  13. MRN- and 9-1-1-Independent Activation of the ATR-Chk1 Pathway during the Induction of the Virulence Program in the Phytopathogen Ustilago maydis.

    Science.gov (United States)

    Tenorio-Gómez, María; de Sena-Tomás, Carmen; Pérez-Martín, Jose

    2015-01-01

    DNA damage response (DDR) leads to DNA repair, and depending on the extent of the damage, to further events, including cell death. Evidence suggests that cell differentiation may also be a consequence of the DDR. During the formation of the infective hypha in the phytopathogenic fungus Ustilago maydis, two DDR kinases, Atr1 and Chk1, are required to induce a G2 cell cycle arrest, which in turn is essential to display the virulence program. However, the triggering factor of DDR in this process has remained elusive. In this report we provide data suggesting that no DNA damage is associated with the activation of the DDR during the formation of the infective filament in U. maydis. We have analyzed bulk DNA replication during the formation of the infective filament, and we found no signs of impaired DNA replication. Furthermore, using RPA-GFP fusion as a surrogate marker of the presence of DNA damage, we were unable to detect any sign of DNA damage at the cellular level. In addition, neither MRN nor 9-1-1 complexes, both instrumental to transmit the DNA damage signal, are required for the induction of the above mentioned cell cycle arrest, as well as for virulence. In contrast, we have found that the claspin-like protein Mrc1, which in other systems serves as scaffold for Atr1 and Chk1, was required for both processes. We discuss possible alternative ways to trigger the DDR, independent of DNA damage, in U. maydis during virulence program activation.

  14. Green synthesis of protein capped silver nanoparticles from phytopathogenic fungus Macrophomina phaseolina (Tassi) Goid with antimicrobial properties against multidrug-resistant bacteria

    Science.gov (United States)

    2014-01-01

    In recent years, green synthesis of nanoparticles, i.e., synthesizing nanoparticles using biological sources like bacteria, algae, fungus, or plant extracts have attracted much attention due to its environment-friendly and economic aspects. The present study demonstrates an eco-friendly and low-cost method of biosynthesis of silver nanoparticles using cell-free filtrate of phytopathogenic fungus Macrophomina phaseolina. UV-visible spectrum showed a peak at 450 nm corresponding to the plasmon absorbance of silver nanoparticles. Scanning electron microscopy (SEM), atomic force microscopy (AFM), and transmission electron microscopy (TEM) revealed the presence of spherical silver nanoparticles of the size range 5 to 40 nm, most of these being 16 to 20 nm in diameter. X-ray diffraction (XRD) spectrum of the nanoparticles exhibited 2θ values corresponding to silver nanoparticles. These nanoparticles were found to be naturally protein coated. SDS-PAGE analysis showed the presence of an 85-kDa protein band responsible for capping and stabilization of the silver nanoparticles. Antimicrobial activities of the silver nanoparticles against human as well as plant pathogenic multidrug-resistant bacteria were assayed. The particles showed inhibitory effect on the growth kinetics of human and plant bacteria. Furthermore, the genotoxic potential of the silver nanoparticles with increasing concentrations was evaluated by DNA fragmentation studies using plasmid DNA. PMID:25114655

  15. Potential of biosynthesized silver nanoparticles using Stenotrophomonas sp. BHU-S7 (MTCC 5978) for management of soil-borne and foliar phytopathogens

    Science.gov (United States)

    Mishra, Sandhya; Singh, Braj Raj; Naqvi, Alim H.; Singh, H. B.

    2017-01-01

    Stenotrophomonas sp. is emerging as a popular microbe of global concern with various potential ecological roles. Biosynthesis of gold and silver nanoparticles (AgNPs) using this bacterial strain has shown promising applications in life sciences. However, there is no report on efficient agricultural applications of biosynthesized AgNPs using Stenotrophomonas sp. In this regard, successful biosynthesis of AgNPs using Stenotrophomonas sp. BHU-S7 (MTCC 5978) was monitored by Uv-visible spectrum showing surface plasmon resonance (SPR) peak at 440 nm. The biosynthesized AgNPs were spherical with an average mean size of ~12 nm. The antifungal efficacy of biosynthesized AgNPs against foliar and soil-borne phytopathogens was observed. The inhibitory impact of AgNPs (2, 4, 10 μg/ml) on conidial germination was recorded under in vitro conditions. Interestingly, sclerotia of Sclerotium rolfsii exposed to AgNPs failed to germinate on PDA medium and in soil system. Moreover, AgNPs treatment successfully managed collar rot of chickpea caused by S. rolfsii under greenhouse conditions. The reduced sclerotia germination, phenolic acids induction, altered lignification and H2O2 production was observed to be the probable mechanisms providing protection to chickpea against S. rolfsii. Our data revealed that AgNPs treated plants are better equipped to cope with pathogen challenge pointing towards their robust applications in plant disease management. PMID:28345581

  16. The high-affinity phosphodiesterase BcPde2 has impact on growth, differentiation and virulence of the phytopathogenic ascomycete Botrytis cinerea.

    Directory of Open Access Journals (Sweden)

    Karin Harren

    Full Text Available Components of the cAMP signaling pathway, such as the adenylate cyclase Bac and the protein kinase A (PKA were shown to affect growth, morphogenesis and differentiation as well as virulence of the phytopathogenic fungus Botrytis cinerea. While loss of Bac caused drastically reduced intracellular cAMP levels, deletion of the PKA resulted in extremely increased cAMP concentrations. To regulate the intracellular level of the second messenger cAMP, a balance between its biosynthesis through adenylate cyclase activity and its hydrolysis by phosphodiesterases (PDEs is crucial. Here, we report the functional characterization of the two PDEs in the ascomycete B. cinerea, BcPde1 and BcPde2. While deletion of bcpde2 resulted in severely affected vegetative growth, conidiation, germination and virulence, the bcpde1 deletion strain displayed a wild-type-like phenotype. However, the double bcpde1/2 deletion mutant exhibited an even stronger phenotype. Localization studies revealed that BcPde2 accumulates at the plasma membrane, but is also localized in the cytoplasm. BcPde1 was shown to be distributed in the cytoplasm as well, but also accumulates in so far unknown mobile vesicles. Overexpression of bcpde1 in the Δbcpde2 background rescued the deletion phenotype, and in addition an increased transcript level of bcpde1 in the Δbcpde2 strain was observed, indicating redundant functions of both PDEs and an interdependent gene expression.

  17. The response of the grape berry moth (Lobesia botrana) to a dietary phytopathogenic fungus (Botrytis cinerea): the significance of fungus sterols.

    Science.gov (United States)

    Mondy; Corio-Costet

    2000-12-01

    A Tortricidae (Lobesia botrana) has a mutualistic relationship with the fungus (Botrytis cinerea). In this study, we investigated the growth, survival, fecundity and amount of sterols and steroids in larvae of this vineyard pest reared on artificial diets containing mycelium (3%) or purified sterols (0.01%) of the phytopathogenic fungus. Two principal questions related to the physiological and biochemical basis of this mutualistic relationship were addressed: (1) how the fungus influences growth, survival, fecundity, sterol and steroid contents of the insect and (2) are fungal sterols involved in the biochemical basis of mutualism? The presence of fungus in the diet led to a decrease of total duration of larval development (mean gain 5.1-9.4 days compared to the total duration in control of 42.9 days), an increase in survival (mean gain 50-76.3%) and fecundity (gain of 94-102%). These positive effects of the fungus on the biology and physiology of the insect were directly correlated to the presence of fungal sterols in the diet. Fungal sterols are one of the biochemical basis of the mutualistic relationship between L. botrana and B. cinerea.

  18. Mobilization of horizontally acquired island 2 is induced in planta in the phytopathogen Pectobacterium atrosepticum SCRI1043 and involves the putative relaxase ECA0613 and quorum sensing.

    Science.gov (United States)

    Vanga, Bhanupratap R; Ramakrishnan, Pavithra; Butler, Ruth C; Toth, Ian K; Ronson, Clive W; Jacobs, Jeanne M E; Pitman, Andrew R

    2015-11-01

    Integrative and conjugative elements (ICEs) contribute to the rapid evolution of bacterial pathogens via horizontal gene transfer of virulence determinants. ICEs have common mechanisms for transmission, yet the cues triggering this process under natural environmental or physiological conditions are largely unknown. In this study, mobilization of the putative ICE horizontally acquired island 2 (HAI2), present in the chromosome of the phytopathogen Pectobacterium atrosepticum SCRI1043, was examined during infection of the host plant potato. Under these conditions, mobilization of HAI2 increased markedly compared with in vitro cultures. In planta-induced mobilization of HAI2 was regulated by quorum sensing and involved the putative ICE-encoded relaxase ECA0613. Disruption of ECA0613 also reduced transcription of genes involved in production of coronafacic acid (Cfa), the major virulence factor harboured on HAI2, whereas their expression was unaffected in the quorum-sensing (expI) mutant. Thus, suppression of cfa gene expression was not regulated by the mobilization of the ICE per se, but was due directly to inactivation of the relaxase. The identification of genetic factors associated solely with in planta mobilization of an ICE demonstrates that this process is highly adapted to the natural environment of the bacterial host and can influence the expression of virulence determinants.

  19. New Protein-Protein Interactions Identified for the Regulatory and Structural Components and Substrates of the Type III Secretion System of the Phytopathogen Xanthomonas axonopodis Pathovar citri

    Science.gov (United States)

    Alegria, Marcos C.; Docena, Cassia; Khater, Leticia; Ramos, Carlos H. I.; da Silva, Ana C. R.; Farah, Chuck S.

    2004-01-01

    We have initiated a project to identify protein-protein interactions involved in the pathogenicity of the bacterial plant pathogen Xanthomonas axonopodis pv. citri. Using a yeast two-hybrid system based on Gal4 DNA-binding and activation domains, we have focused on identifying interactions involving subunits, regulators, and substrates of the type III secretion system coded by the hrp (for hypersensitive response and pathogenicity), hrc (for hrp conserved), and hpa (for hrp associated) genes. We have identified several previously uncharacterized interactions involving (i) HrpG, a two-component system response regulator responsible for the expression of X. axonopodis pv. citri hrp operons, and XAC0095, a previously uncharacterized protein encountered only in Xanthomonas spp.; (ii) HpaA, a protein secreted by the type III secretion system, HpaB, and the C-terminal domain of HrcV; (iii) HrpB1, HrpD6, and HrpW; and (iv) HrpB2 and HrcU. Homotropic interactions were also identified for the ATPase HrcN. These newly identified protein-protein interactions increase our understanding of the functional integration of phytopathogen-specific type III secretion system components and suggest new hypotheses regarding the molecular mechanisms underlying Xanthomonas pathogenicity. PMID:15342589

  20. Tallow amphopolycarboxyglycinate-stabilized silver nanoparticles: new frontiers in development of plant protection products with a broad spectrum of action against phytopathogens

    Science.gov (United States)

    Krutyakov, Yurii A.; Kudrinskiy, Alexey A.; Zherebin, Pavel M.; Yapryntsev, Alexey D.; Pobedinskaya, Marina A.; Elansky, Sergey N.; Denisov, Albert N.; Mikhaylov, Dmitry M.; Lisichkin, Georgii V.

    2016-07-01

    Sustainable agriculture calls for minimal use of agrochemicals in order to protect the environment. It has caused an increase in the rate of nanoparticles use, in particular silver nanoparticles (AgNPs) due to their safety for mammals, unique biological activity and a broad spectrum of action against fungal and bacterial pathogens. Until now the use of AgNPs dispersions in the agricultural sector has been essentially limited due to many factors decreased their stability (mixing with other pesticides, presence of electrolytes). We present a versatile synthesis of polyampholyte surfactant (tallow amphopolycarboxyglycinate) stabilized AgNPs. We took a close look at unique aggregation behavior (via dynamic light scattering and UV-vis spectroscopy) and biocidal activity of obtained silver colloids. AgNPs are characterized by exclusively high aggregative stability in the presence of coagulating agents NaNO3 and NaSO4 (up to 1 M), during drying/redispergation, and frost/defrost cycles. The dispersion of AgNPs shows high biocidal activity (EC50 is ten times lower than commercial species ones) with respect to Phytophthora infestans and phytopathogenic fungi. This points to the possibility of successful application of silver preparations within agriculture with the goal of partial reduction of the use of toxic and expensive synthetic antibiotics and pesticides.

  1. Environmental Bacteriophages of the Emerging Enterobacterial Phytopathogen, Dickeya solani, Show Genomic Conservation and Capacity for Horizontal Gene Transfer between Their Bacterial Hosts

    Directory of Open Access Journals (Sweden)

    Andrew Day

    2017-08-01

    Full Text Available Dickeya solani is an economically important phytopathogen widespread in mainland Europe that can reduce potato crop yields by 25%. There are no effective environmentally-acceptable chemical systems available for diseases caused by Dickeya. Bacteriophages have been suggested for use in biocontrol of this pathogen in the field, and limited field trials have been conducted. To date only a small number of bacteriophages capable of infecting D. solani have been isolated and characterized, and so there is a need to expand the repertoire of phages that may have potential utility in phage therapy strategies. Here we describe 67 bacteriophages from environmental sources, the majority of which are members of the viral family Myoviridae. Full genomic sequencing of two isolates revealed a high degree of DNA identity with D. solani bacteriophages isolated in Europe in the past 5 years, suggesting a wide ecological distribution of this phage family. Transduction experiments showed that the majority of the new environmental bacteriophages are capable of facilitating efficient horizontal gene transfer. The possible risk of unintentional transfer of virulence or antibiotic resistance genes between hosts susceptible to transducing phages cautions against their environmental use for biocontrol, until specific phages are fully tested for transduction capabilities.

  2. Strigolactone biosynthesis is evolutionarily conserved, regulated by phosphate starvation and contributes to resistance against phytopathogenic fungi in a moss, Physcomitrella patens

    KAUST Repository

    Decker, Eva L.

    2017-03-06

    In seed plants, strigolactones (SLs) regulate architecture and induce mycorrhizal symbiosis in response to environmental cues. SLs are formed by combined activity of the carotenoid cleavage dioxygenases (CCDs) 7 and 8 from 9-cis-β-carotene, leading to carlactone that is converted by cytochromes P450 (clade 711; MAX1 in Arabidopsis) into various SLs. As Physcomitrella patens possesses CCD7 and CCD8 homologs but lacks MAX1, we investigated if PpCCD7 together with PpCCD8 form carlactone and how deletion of these enzymes influences growth and interactions with the environment. We investigated the enzymatic activity of PpCCD7 and PpCCD8 in vitro, identified the formed products by high performance liquid chromatography (HPLC) and LC-MS, and generated and analysed ΔCCD7 and ΔCCD8 mutants. We defined enzymatic activity of PpCCD7 as a stereospecific 9-cis-CCD and PpCCD8 as a carlactone synthase. ΔCCD7 and ΔCCD8 lines showed enhanced caulonema growth, which was revertible by adding the SL analogue GR24 or carlactone. Wild-type (WT) exudates induced seed germination in Orobanche ramosa. This activity was increased upon phosphate starvation and abolished in exudates of both mutants. Furthermore, both mutants showed increased susceptibility to phytopathogenic fungi. Our study reveals the deep evolutionary conservation of SL biosynthesis, SL function, and its regulation by biotic and abiotic cues.

  3. Molecular characterization of CLPT1, a SEC4-like Rab/GTPase of the phytopathogenic fungus Colletotrichum lindemuthianum which is regulated by the carbon source.

    Science.gov (United States)

    Dumas, B; Borel, C; Herbert, C; Maury, J; Jacquet, C; Balsse, R; Esquerré-Tugayé, M T

    2001-07-11

    The gene CLPT1 (Colletotrichum lindemuthianum Protein Transport 1) encoding a Rab/GTPase was isolated from the filamentous fungus Colletotrichum lindemuthianum, the causal agent of bean anthracnose. At the amino acid level, CLPT1 shows between 54 and 80% identity to SEC4-like proteins, a class of molecules required for intracellular vesicular transport in yeasts. In particular, typical SEC4 domains involved in nucleotide binding and membrane attachment are present in the CLPT1 sequence. Functional identity of CLPT1 with SEC4 was confirmed by complementation of the Saccharomyces cerevisiae sec4-8 mutation. This is the first report of a gene involved in the control of intracellular vesicular trafficking in a phytopathogenic fungus. RNA blot analyses of CLPT1 expression were performed during in vitro growth of the fungus on synthetic media containing glucose or pectin, as single carbon source. The accumulation of CLPT1 mRNA was strongly increased on pectin, a plant cell wall polysaccharide that induces the production of extracellular pectinases, whereas the level of CLPT1 mRNA was below the detection threshold on glucose. These results suggest that CLPT1 is mainly involved in protein secretion and that the production of extracellular enzymes potentially involved in pathogenesis in filamentous fungi is sustained by induction of the genes involved in the secretory machinery.

  4. 黄栌枯萎病菌拮抗细菌的分离与鉴定%Isolation and identification of an antagonistic bacterium against smoke tree wilt fungus Verticillium dahliae

    Institute of Scientific and Technical Information of China (English)

    孙妍; 于地美; 黄华毅; 王永林; 田呈明

    2015-01-01

    为了获得能有效拮抗黄栌枯萎病的大丽轮枝菌( Verticillium dahliae)和尖孢镰刀菌( Fusarium oxysporum)的生防菌株,采用系列稀释分离法和平板对峙法从盐碱地土壤中筛选拮抗细菌,基于形态特征、生理生化特性和16S rDNA序列分析鉴定菌株,并测定其无菌滤液和挥发性气体的抑菌活性。筛选结果表明,菌株C-2-3-2是一株能有效抑制黄栌枯萎病菌和尖孢镰刀菌的生防菌株,经鉴定其为枯草芽孢杆菌( Bacillus subtilis)。该菌株的无菌滤液对黄栌枯萎病菌和尖孢镰刀菌均具有明显的体外抑菌作用,且能有效抑制黄栌枯萎病菌微菌核的形成。同时,其挥发性气体也能有效抑制黄栌枯萎病菌的生长。另外,研究还发现该菌株能产生铁载体和合成氨,并产生几丁质酶、纤维素酶、蛋白酶和β-1,3-葡聚糖酶。研究结果表明菌株C-2-3-2在黄栌枯萎病生物防治中具有较好的应用潜力和较高的研究价值。%In order to isolate and characterize an antagonistic bacterial strain against Verticillium dahliae and Fusarium oxysporum, serial dilution and plate⁃confrontation methods were used for preliminary screening and selecting antagonistic bacteria from saline⁃alkali soil. Strain identification was based on morphological and physiological characteristics, and phylogenetic analysis of 16S rDNA sequence. The antagonistic effects of sterile culture filtrates and volatiles of antagonis⁃tic bacteria were determined. An antagonistic bacteria, named as C-2-3-2, was isolated and identified as Bacillus sub⁃tilis. The results indicated that sterile culture filtrate of the strain exhibited obvious in vitro inhibition against V. dahliae and F. oxysporum, and suppressed the production of microsclerotia of V. dahliae. The volatiles also effectively inhibited the mycelial growth of V. dahliae. In addition, the results indicated that C-2

  5. Eukaryotic extracellular catalase–peroxidase from Magnaporthe grisea – Biophysical/chemical characterization of the first representative from a novel phytopathogenic KatG group

    Science.gov (United States)

    Zámocký, Marcel; Droghetti, Enrica; Bellei, Marzia; Gasselhuber, Bernhard; Pabst, Martin; Furtmüller, Paul G.; Battistuzzi, Gianantonio; Smulevich, Giulietta; Obinger, Christian

    2012-01-01

    All phytopathogenic fungi have two catalase–peroxidase paralogues located either intracellularly (KatG1) or extracellularly (KatG2). Here, for the first time a secreted bifunctional, homodimeric catalase–peroxidase (KatG2 from the rice blast fungus Magnaporthe grisea) has been produced heterologously with almost 100% heme occupancy and comprehensively investigated by using a broad set of methods including UV–Vis, ECD and resonance Raman spectroscopy (RR), thin-layer spectroelectrochemistry, mass spectrometry, steady-state & presteady-state spectroscopy. RR spectroscopy reveals that MagKatG2 shows a unique mixed-spin state, non-planar heme b, and a proximal histidine with pronounced imidazolate character. At pH 7.0 and 25 °C, the standard reduction potential E°′ of the Fe(III)/Fe(II) couple for the high-spin native protein was found to fall in the range typical for the KatG family. Binding of cyanide was relatively slow at pH 7.0 and 25 °C and with a Kd value significantly higher than for the intracellular counterpart. Demonstrated by mass spectrometry MagKatG2 has the typical Trp118-Tyr251-Met277 adduct that is essential for its predominantly catalase activity at the unique acidic pH optimum. In addition, MagKatG2 acts as a versatile peroxidase using both one- and two-electron donors. Based on these data, structure–function relationships of extracellular eukaryotic KatGs are discussed with respect to intracellular KatGs and possible role(s) in host–pathogen interaction. PMID:21971530

  6. Eukaryotic extracellular catalase-peroxidase from Magnaporthe grisea - Biophysical/chemical characterization of the first representative from a novel phytopathogenic KatG group.

    Science.gov (United States)

    Zámocký, Marcel; Droghetti, Enrica; Bellei, Marzia; Gasselhuber, Bernhard; Pabst, Martin; Furtmüller, Paul G; Battistuzzi, Gianantonio; Smulevich, Giulietta; Obinger, Christian

    2012-03-01

    All phytopathogenic fungi have two catalase-peroxidase paralogues located either intracellularly (KatG1) or extracellularly (KatG2). Here, for the first time a secreted bifunctional, homodimeric catalase-peroxidase (KatG2 from the rice blast fungus Magnaporthe grisea) has been produced heterologously with almost 100% heme occupancy and comprehensively investigated by using a broad set of methods including UV-Vis, ECD and resonance Raman spectroscopy (RR), thin-layer spectroelectrochemistry, mass spectrometry, steady-state & presteady-state spectroscopy. RR spectroscopy reveals that MagKatG2 shows a unique mixed-spin state, non-planar heme b, and a proximal histidine with pronounced imidazolate character. At pH 7.0 and 25 °C, the standard reduction potential E°' of the Fe(III)/Fe(II) couple for the high-spin native protein was found to fall in the range typical for the KatG family. Binding of cyanide was relatively slow at pH 7.0 and 25 °C and with a K(d) value significantly higher than for the intracellular counterpart. Demonstrated by mass spectrometry MagKatG2 has the typical Trp118-Tyr251-Met277 adduct that is essential for its predominantly catalase activity at the unique acidic pH optimum. In addition, MagKatG2 acts as a versatile peroxidase using both one- and two-electron donors. Based on these data, structure-function relationships of extracellular eukaryotic KatGs are discussed with respect to intracellular KatGs and possible role(s) in host-pathogen interaction.

  7. MRN- and 9-1-1-Independent Activation of the ATR-Chk1 Pathway during the Induction of the Virulence Program in the Phytopathogen Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    María Tenorio-Gómez

    Full Text Available DNA damage response (DDR leads to DNA repair, and depending on the extent of the damage, to further events, including cell death. Evidence suggests that cell differentiation may also be a consequence of the DDR. During the formation of the infective hypha in the phytopathogenic fungus Ustilago maydis, two DDR kinases, Atr1 and Chk1, are required to induce a G2 cell cycle arrest, which in turn is essential to display the virulence program. However, the triggering factor of DDR in this process has remained elusive. In this report we provide data suggesting that no DNA damage is associated with the activation of the DDR during the formation of the infective filament in U. maydis. We have analyzed bulk DNA replication during the formation of the infective filament, and we found no signs of impaired DNA replication. Furthermore, using RPA-GFP fusion as a surrogate marker of the presence of DNA damage, we were unable to detect any sign of DNA damage at the cellular level. In addition, neither MRN nor 9-1-1 complexes, both instrumental to transmit the DNA damage signal, are required for the induction of the above mentioned cell cycle arrest, as well as for virulence. In contrast, we have found that the claspin-like protein Mrc1, which in other systems serves as scaffold for Atr1 and Chk1, was required for both processes. We discuss possible alternative ways to trigger the DDR, independent of DNA damage, in U. maydis during virulence program activation.

  8. Aspergillus piperis A/5 from plum-distilling waste compost produces a complex of antifungal metabolites active against the phytopathogen Pythium aphanidermatum

    Directory of Open Access Journals (Sweden)

    Jovičić-Petrović Jelena

    2016-01-01

    Full Text Available Adding compost to soil can result in plant disease suppression through the mechanisms of antagonistic action of compost microflora against plant pathogens. The aim of the study was to select effective antagonists of Pythium aphanidermatum from compost, to assess the effect of its extracellular metabolites on the plant pathogen, and to characterize antifungal metabolites. The fungal isolate selected by a confrontation test was identified as Aspergillus piperis A/5 on the basis of morphological features and the internal transcribed spacer (ITS region, β-tubulin and calmodulin partial sequences. Liquid chromatography-mass spectroscopy (LC-MS analysis showed that gluconic and citric acid were the most abundant in the organic culture extract. However, the main antifungal activity was contained in the aqueous phase remaining after the organic solvent extraction. The presence of considerable amounts of proteins in both the crude culture extract as well as the aqueous phase remaining after solvent extraction was confirmed by SDS-PAGE. Isolated Aspergillus piperis A/5 exhibits strong antifungal activity against the phytopathogen Pythium aphanidermatum. It secretes a complex mixture of metabolites consisting of small molecules, including gluconic acid, citric acid and itaconic acid derivatives, but the most potent antifungal activity was associated with proteins resistant to heat and organic solvents. Our findings about the activity and characterization of antagonistic strain metabolites contribute to the understanding of the mechanism of interaction of antifungal metabolites as well as fungal-fungal interaction. The obtained results provide a basis for further application development in agriculture and food processing. [Projekat Ministarstva nauke Republike Srbije, br. TR31080 i br. ON173048, i EU Commission project AREA, br.316004

  9. Comprehensive overexpression analysis of cyclic-di-GMP signalling proteins in the phytopathogen Pectobacterium atrosepticum reveals diverse effects on motility and virulence phenotypes.

    Science.gov (United States)

    Tan, H; West, J A; Ramsay, J P; Monson, R E; Griffin, J L; Toth, I K; Salmond, G P C

    2014-07-01

    Bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) is a ubiquitous bacterial signalling molecule produced by diguanylate cyclases of the GGDEF-domain family. Elevated c-di-GMP levels or increased GGDEF protein expression is frequently associated with the onset of sessility and biofilm formation in numerous bacterial species. Conversely, phosphodiesterase-dependent diminution of c-di-GMP levels by EAL- and HD-GYP-domain proteins is often accompanied by increased motility and virulence. In this study, we individually overexpressed 23 predicted GGDEF, EAL or HD-GYP-domain proteins encoded by the phytopathogen Pectobacterium atrosepticum strain SCRI1043. MS-based detection of c-di-GMP and 5'-phosphoguanylyl-(3'-5')-guanosine in these strains revealed that overexpression of most genes promoted modest 1-10-fold changes in cellular levels of c-di-GMP, with the exception of the GGDEF-domain proteins ECA0659 and ECA3374, which induced 1290- and 7660-fold increases, respectively. Overexpression of most EAL domain proteins increased motility, while overexpression of most GGDEF domain proteins reduced motility and increased poly-β-1,6-N-acetyl-glucosamine-dependent flocculation. In contrast to domain-based predictions, overexpression of the EAL protein ECA3549 or the HD-GYP protein ECA3548 increased c-di-GMP concentrations and reduced motility. Most overexpression constructs altered the levels of secreted cellulases, pectinases and proteases, confirming c-di-GMP regulation of virulence in Pe. atrosepticum. However, there was no apparent correlation between virulence-factor induction and the domain class expressed or cellular c-di-GMP levels, suggesting that regulation was in response to specific effectors within the network, rather than total c-di-GMP concentration. Finally, we demonstrated that the cellular localization patterns vary considerably for GGDEF/EAL/HD-GYP proteins, indicating it is a likely factor restricting specific interactions within the c

  10. A novel single-stranded RNA virus isolated from a phytopathogenic filamentous fungus, Rosellinia necatrix, with similarity to hypo-like viruses

    Directory of Open Access Journals (Sweden)

    Rui eZhang

    2014-07-01

    Full Text Available Here we report a biological and molecular characterization of a novel positive-sense RNA virus isolated from a field isolate (NW10 of a filamentous phytopathogenic fungus, the white root rot fungus that is designated as Rosellinia necatrix fusarivirus 1 (RnFV1. A recently developed technology using zinc ions allowed us to transfer RnFV1 to two mycelially incompatible Rosellinia necatrix strains. A biological comparison of the virus-free and -recipient isogenic fungal strains suggested that RnFV1 infects latently and thus has no potential as a virocontrol agent. The virus has an undivided positive-sense RNA genome of 6286 nucleotides excluding a poly (A tail. The genome possesses two non-overlapping open reading frames (ORFs: a large ORF1 that encodes polypeptides with RNA replication functions and a smaller ORF2 that encodes polypeptides of unknown function. A lack of coat protein genes was suggested by the failure of virus particles from infected mycelia. No evidence was obtained by Northern analysis or classical 5'-RACE for the presence of subgenomic RNA for the downstream ORF. Sequence similarities were found in amino-acid sequence between RnFV1 putative proteins and counterparts of a previously reported mycovirus, Fusarium graminearum virus 1 (FgV1. Interestingly, several related sequences were detected by BLAST searches of independent transcriptome assembly databases one of which probably represents an entire virus genome. Phylogenetic analysis based on the conserved RNA-dependent RNA polymerase showed that RnFV1, FgV1, and these similar sequences are grouped in a cluster distinct from distantly related hypoviruses. It is proposed that a new taxonomic family termed Fusariviridae be created to include RnFV1and FgV1.

  11. Ação antagônica e de metabólitos bioativos de Trichoderma spp. contra os patógenos Sclerotium rolfsii e Verticillium dahliae

    Directory of Open Access Journals (Sweden)

    Carolina Oliveira Isaias

    2014-03-01

    Full Text Available O efeito antagônico exercido por fungos do gênero Trichoderma foi avaliado contra os fitopatógenos Sclerotium rolfsii e Verticillium dahliae, isolados de áster ornamental e morangueiro, respectivamente. Utilizaram-se 20 isolados do antagonista, pertencentes às espécies T. asperellum, T. hamatum, T. harzianum, T. koningiopsis, T. spirale e Trichoderma sp. O antagonismo foi verificado por meio de testes de culturas pareadas e produção de metabólitos bioativos pelo método de sobreposição de placas. A maioria dos isolados exibiu efeito inibitório sobre o crescimento micelial de S. rolfsii e V. dahliae quando comparado com as testemunhas. Excetuando-se os testes de produção de metabólitos não-voláteis, verificou-se maior ação inibitória contra V. dahliae. Mesmo os metabólitos não-voláteis termoestáveis apresentaram efeito inibitório contra ambos os patógenos testados, mostrando que os metabólitos secundários do antagonista exercem efeito significativo sobre essas espécies patogênicas.

  12. A primary study on effec of VAM fungi to strawberry and Verticillium wilt%VAM真菌对草莓促生、防草莓黄萎病效应初探

    Institute of Scientific and Technical Information of China (English)

    马宝红; 甄文超; 曹克强; 武玉环

    2004-01-01

    在室内条件下研究了Glomus mosseae和Glomus versiforme与草莓共生后对由大丽轮枝菌(Verticillium dahliae Kelb)引起的草莓黄萎病的抗病效果.结果表明:两种VAM真菌对草莓黄萎病均有明显的防效.与对照相比接种G.mosseae和G.versiforme的处理病情指数分别降低39.4和34.7,防效分别为66.22%和58.32%.此外,两种VAM真菌对草莓生长有明显的促进作用,接种G.mosseae和G.versiforme处理的单株株高、茎粗、根长和地上部鲜重等各项指标均明显高于对照.

  13. Idelntification and fermentation condition for Verticillium lecanii with high nematicidal activity%高杀线活性蜡蚧轮枝菌鉴定与发酵条件优化研究

    Institute of Scientific and Technical Information of China (English)

    孙艳; 段玉玺; 陈立杰

    2009-01-01

    通过室内试验,筛选得到1株具有高杀线活性的菌株Snef69,其发酵液的60%稀释液对南方根结线虫(Meloidogyne incognita)J2的校正致死率达到91.27%.通过ITS及形态学鉴定,并测定了不同发酵条件下Snef69菌株对南方根结线虫的毒力作用效果,结果表明该菌株为蜡蚧轮枝菌(Verticillium lecanii),随着发酵液浓度的增加,杀线活性增强,当发酵条件中碳源为麦芽糖、氮源为NaNO3、发酵天数为6~7 d、pH值=6时杀线虫效果最明显.

  14. 枯草芽孢杆菌植入棉苗体内拮抗黄萎病研究%The Studies of Bacillus Subtilis Implanted in Cotton Breedings to Antagonize Verticillium Wilt

    Institute of Scientific and Technical Information of China (English)

    王谧; 王维东; 范玉刚

    2011-01-01

    在棉花无土育苗基质中植入枯草芽孢杆菌,使其优先占领生态位,而拮抗黄萎病。结果表明:枯草芽孢杆菌CD102菌液稀释后植入灭菌基质,能在短期内增殖和持续繁殖;采用浸种、浇灌、针刺和喷雾的方法处理基质培育的棉苗,CD102均能不同程度定植其中;带CD102的基质培育棉苗,移栽大田后对黄萎病具有较好防治效果。%Bacillus subtilis CD102 was implanted to cotton seedling substrate,It had priority to occupy ecological niche and antagonized the infection of Verticillium Wilt.Results showed that: the bacillus subtilis CD102 was implanted to sterilized substrate and proliferated rapidly and continuously;It was all implanted to cotton seedling raised by substance,which accomplished by all methods including seed-soaking,pouring,acupuncture and spraying.Cotton seedlings raised by substrate with CD102 had good antagonistic effect to Vertillium Wilt after transplanting to field.

  15. Bio-fabrication of silver nanoparticles using the leaf extract of an ancient herbal medicine, dandelion (Taraxacum officinale), evaluation of their antioxidant, anticancer potential, and antimicrobial activity against phytopathogens.

    Science.gov (United States)

    Saratale, Rijuta G; Benelli, Giovanni; Kumar, Gopalakrishnan; Kim, Dong Su; Saratale, Ganesh D

    2017-07-11

    In recent years, the use of nanoparticle-based antimicrobials has been increased due to many advantages over conventional agrochemicals. This study investigates the utilization of common medicinal plant dandelion, Taraxacum officinale, for the synthesis of silver nanoparticles (TOL-AgNPs). AgNPs were evaluated for antimicrobial activity against two important phytopathogens, Xanthomonas axonopodis and Pseudomonas syringae. The morphology, size, and structure of TOL-AgNPs were characterized using UV-visible spectroscopy and X-ray diffraction (XRD). Fourier transform infrared spectroscopy (FT-IR) showed the presence of phytochemicals involved during synthesis of NPs. High-resolution transmission electron microscopy (HR-TEM) analysis shed light on the size of monodispersed spherical AgNPs ranging between 5 and 30 nm, with an average particle size of about 15 nm. The TOL-AgNPs (at 20 μg/mL concentration) showed significant antibacterial activity with significant growth inhibition of phytopathogens X. axonopodis (22.0 ± 0.84 mm) and P. syringae (19.5 ± 0.66 mm). The synthesized AgNPs had higher antibacterial activity in comparison with commercial AgNPs. Synergistic assays with standard antibiotics revealed that nanoformulations with tetracycline showed better broad-spectrum efficiency to control phytopathogens. They also possessed significant antioxidant potential in terms of ABTS (IC50 = 45.6 μg/mL), DPPH (IC50 = 56.1 μg/mL), and NO (IC50 = 55.2 μg/mL) free radical scavenging activity. The TOL-AgNPs showed high cytotoxic effect against human liver cancer cells (HepG2). Overall, dandelion-mediated AgNPs synthesis can represent a novel approach to develop effective antimicrobial and anticancer drugs with a cheap and eco-friendly nature.

  16. A Facile and Efficient Synthesis of Diaryl Amines or Ethers under Microwave Irradiation at Presence of KF/Al2O3 without Solvent and Their Anti-Fungal Biological Activities against Six Phytopathogens

    Directory of Open Access Journals (Sweden)

    Tao Zhang

    2013-09-01

    Full Text Available A series of diaryl amines, ethers and thioethers were synthesized under microwave irradiation efficiently at presence of KF/Al2O3 in 83%–96% yields without any solvent. The salient characters of this method lie in short reaction time, high yields, general applicability to substrates and simple workup procedure. At the same time, their antifungal biological activities against six phytopathogen were evaluated. Most of the compounds (3b, 3c, 3g–o are more potent than thiophannate-methyl against to Magnaporthe oryzae. This implies that diaryl amine or ether moiety may be helpful in finding a fungicide against Magnaporthe oryzae.

  17. ldentification and Control Methods of Eggplant Southern Blight, Damp-ing-off, Root Rot, Verticil-lium Wilt and Fusarium Wilt%茄子白绢病、猝倒病、根腐病、黄萎病和枯萎病的识别与防治

    Institute of Scientific and Technical Information of China (English)

    高焕; 郑丹丹

    2012-01-01

    介绍了茄子白绢病、猝倒病、根腐病、黄萎病和枯萎病的危害症状、发病特点,并从农业措施、化学防治等方面总结了各病害的综合防治方法。%The symptom and disease characteristics of eggplant southern blight, damping-off, root rot, verticillium wilt and fusarium wilt were introduced. Then the integrated control methods were put forward, which contained measures of agricultural control, chemical control and so on.

  18. Isolation, characterization and molecular three-dimensional structural predictions of metalloprotease from a phytopathogenic fungus, Alternaria solani (Ell. and Mart.) Sor.

    Science.gov (United States)

    Chandrasekaran, Murugesan; Chandrasekar, Raman; Chun, Se-Chul; Sathiyabama, Muthukrishnan

    2016-08-01

    The present study aims at isolation, identification, characterization and prediction of three-dimensional molecular architecture of a proteolytic enzyme from the early blight pathogen, Alternaria solani which are hypothesized to be a marker of phytopathogenicity. Maximum enzyme production by A. solani was observed in Czapex's Dox broth amended with 2% (w/v) casein than other inducer amendments. Results indicate that the enzyme remained highly active in a pH range of 7.0-10.0 and a temperature range of 45-50°C. The enzyme was strongly inhibited by EDTA, whereas phenylmethylsulfonyl fluoride and monovalent cations (Na(+), K(+)) had little effect. Metal ions such as MgSO4, CaCl2, KCl at 10 mM concentration showed a stimulatory effect (>85%) on protease activity. Matrix-assisted laser desorption and ionization time of flight/mass spectrometry analysis of partially purified enzyme revealed the presence of protease belonging to a keratinolytic protein (metalloprotease) of exopeptidase nature. Putative A. solani keratinolytic enzyme (AsK) is made up of 216 amino acid residues with molecular weight (MW) 24.5 kDa, having a molecular formula of C1094H1704N290O342S4. Ramachandran plot analysis of the protein residues falling into the most favored secondary structures was observed at 84.2%. The major protein structural blocks, 2-β-sheets, and 9-α-helices have a greater tendency to be conserved during the evolutionary process than do mere sequences of amino acids. Besides, AsK, model prediction showed the presence of a Zinc atom at helix regions (Helix 3, 6, 7: His(57), His(130), His(169), and Cys(123)). Thus, it can be concluded that the major proteinases of AsK are divalent cation-requiring metalloproteinases and make them potential targets of protease inhibitors designing. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  19. Antibiosis and dark-pigments secretion by the phytopathogenic and environmental fungal species after interaction in vitro with a Bacillus subtilis isolate

    Directory of Open Access Journals (Sweden)

    Alexandre Paulo Machado

    2010-10-01

    Full Text Available In this work, different reactions in vitro between an environmental bacterial isolate and fungal species were related. The Gram-positive bacteria had terminal and subterminal endospores, presented metabolic characteristics of mesophilic and acidophilic growth, halotolerance, positive to nitrate reduction and enzyme production, as caseinase and catalase. The analysis of partial sequences containing 400 to 700 bases of the 16S ribosomal RNA gene showed identity with the genus Bacillus. However, its identity as B. subtilis was confirmed after analyses of the rpoB, gyrA, and 16S rRNA near-full-length sequences. Strong inhibitory activity of environmental microorganisms, such as Penicillium sp, Aspergillus flavus, A. niger, and phytopathogens, such as Colletotrichum sp, Alternaria alternata, Fusarium solani and F. oxysporum f.sp vasinfectum, was shown on co-cultures with B. subtilis strain, particularly on Sabouraud dextrose agar (SDA and DNase media. Red and red-ochre color pigments, probably phaeomelanins, were secreted by A. alternata and A. niger respectively after seven days of co-culture.Na presente investigação, nosso objetivo principal foi relatar diferentes interações in vitro de um isolado bacteriano ambiental com espécies fúngicas. Através da identificação clássica, nós verificamos que o bacilo ambiental apresentava endósporos terminais e subterminais, características metabólicas de mesofilia, acidofilia, halotolerância, redução de nitrato e produção de enzimas, como caseinase e catalase. Análise de seqüências parciais do gene 16S RNAr contendo de 400 a 700 bases revelou identidade com gênero Bacillus. No entanto, a espécie Bacillus subtilis foi confirmada somente depois da análise de seqüências dos genes rpoB, gyrA, and 16S RNAr. Intensa atividade inibitória aos fungos ambientais, como Penicillium sp, Aspergillus flavus, A. niger, e fitopatogênicos, como Colletotrichum sp, Alternaria alternata, Fusarium solani

  20. Cloning and characterization of a pectin lyase gene from Colletotrichum lindemuthianum and comparative phylogenetic/structural analyses with genes from phytopathogenic and saprophytic/opportunistic microorganisms.

    Science.gov (United States)

    Lara-Márquez, Alicia; Zavala-Páramo, María G; López-Romero, Everardo; Calderón-Cortés, Nancy; López-Gómez, Rodolfo; Conejo-Saucedo, Ulises; Cano-Camacho, Horacio

    2011-12-09

    Microorganisms produce cell-wall-degrading enzymes as part of their strategies for plant invasion/nutrition. Among these, pectin lyases (PNLs) catalyze the depolymerization of esterified pectin by a β-elimination mechanism. PNLs are grouped together with pectate lyases (PL) in Family 1 of the polysaccharide lyases, as they share a conserved structure in a parallel β-helix. The best-characterized fungal pectin lyases are obtained from saprophytic/opportunistic fungi in the genera Aspergillus and Penicillium and from some pathogens such as Colletotrichum gloeosporioides.The organism used in the present study, Colletotrichum lindemuthianum, is a phytopathogenic fungus that can be subdivided into different physiological races with different capacities to infect its host, Phaseolus vulgaris. These include the non-pathogenic and pathogenic strains known as races 0 and 1472, respectively. Here we report the isolation and sequence analysis of the Clpnl2 gene, which encodes the pectin lyase 2 of C. lindemuthianum, and its expression in pathogenic and non-pathogenic races of C. lindemuthianum grown on different carbon sources. In addition, we performed a phylogenetic analysis of the deduced amino acid sequence of Clpnl2 based on reported sequences of PNLs from other sources and compared the three-dimensional structure of Clpnl2, as predicted by homology modeling, with those of other organisms. Both analyses revealed an early separation of bacterial pectin lyases from those found in fungi and oomycetes. Furthermore, two groups could be distinguished among the enzymes from fungi and oomycetes: one comprising enzymes from mostly saprophytic/opportunistic fungi and the other formed mainly by enzymes from pathogenic fungi and oomycetes. Clpnl2 was found in the latter group and was grouped together with the pectin lyase from C. gloeosporioides. The Clpnl2 gene of C. lindemuthianum shares the characteristic elements of genes coding for pectin lyases. A time-course analysis

  1. Cloning and characterization of a pectin lyase gene from Colletotrichum lindemuthianum and comparative phylogenetic/structural analyses with genes from phytopathogenic and saprophytic/opportunistic microorganisms

    Directory of Open Access Journals (Sweden)

    Lara-Márquez Alicia

    2011-12-01

    Full Text Available Abstract Background Microorganisms produce cell-wall-degrading enzymes as part of their strategies for plant invasion/nutrition. Among these, pectin lyases (PNLs catalyze the depolymerization of esterified pectin by a β-elimination mechanism. PNLs are grouped together with pectate lyases (PL in Family 1 of the polysaccharide lyases, as they share a conserved structure in a parallel β-helix. The best-characterized fungal pectin lyases are obtained from saprophytic/opportunistic fungi in the genera Aspergillus and Penicillium and from some pathogens such as Colletotrichum gloeosporioides. The organism used in the present study, Colletotrichum lindemuthianum, is a phytopathogenic fungus that can be subdivided into different physiological races with different capacities to infect its host, Phaseolus vulgaris. These include the non-pathogenic and pathogenic strains known as races 0 and 1472, respectively. Results Here we report the isolation and sequence analysis of the Clpnl2 gene, which encodes the pectin lyase 2 of C. lindemuthianum, and its expression in pathogenic and non-pathogenic races of C. lindemuthianum grown on different carbon sources. In addition, we performed a phylogenetic analysis of the deduced amino acid sequence of Clpnl2 based on reported sequences of PNLs from other sources and compared the three-dimensional structure of Clpnl2, as predicted by homology modeling, with those of other organisms. Both analyses revealed an early separation of bacterial pectin lyases from those found in fungi and oomycetes. Furthermore, two groups could be distinguished among the enzymes from fungi and oomycetes: one comprising enzymes from mostly saprophytic/opportunistic fungi and the other formed mainly by enzymes from pathogenic fungi and oomycetes. Clpnl2 was found in the latter group and was grouped together with the pectin lyase from C. gloeosporioides. Conclusions The Clpnl2 gene of C. lindemuthianum shares the characteristic elements of

  2. 棉花黄萎病拮抗细菌HMB-1005鉴定及定殖%Studies on identification and colonization of antagonistic bacteria HMB-1005 to cotton Verticillium dahliae

    Institute of Scientific and Technical Information of China (English)

    郭慧娟; 李术娜; 李红亚; 王全; 王树香; 王伟; 朱宝成

    2011-01-01

    In this study, determination of antagonistic activity was done to the strain HMB-1005 from the cotton fields with the Verticillium dahliae strains as the instructions, characteristics of strain HMB-1005 equates well with Bacillus amyloliquefaciens at the level of species in morphological, 19 physiological biochemical items and 16S rDNA sequence. This test required sterile soil and non-sterile soil in two soil samples, this test need two soil samples of three different treatment that was: The antagonistic bacterium bacillus soaking liquid, all mixed soil and part of soil for plants mixed. The purpose of this paper was to study antagonistic bacteria HMB-1005 in the soil and in the colonization of cotton roots and rhizosphere colonization of the situation. The results showed that antagonistic bacteria HMB-1005 in non-sterilized soil and sterilized soil colonization could reproduce, and the quantity achieved 106cfu/g. Through revovery bacteria of cotton roots, confirmed the antagonistic bacteria in the HMB-1005 designated cotton root colonization, number of 103cfu/g. This experiment indicated the antibacterial number in antiseptic was higher than the non-antiseptic earth, and the difference during three processing modes was not obviously. Considered the various factor, minded that the first method would be better.%以大丽轮枝菌(Verticillium dahliae)为指示菌,对从棉田土壤中已经分离筛选到的细菌HMB-1005进行形态、19项生理生化特征及16S rDNA试验,结果表明,HMB-1005为解淀粉芽孢杆菌(Bacillus amyloliquefaciens);通过灭菌土和非灭菌土进行盆栽试验,拮抗菌HMB-1005芽孢液浸种、土壤全部混菌和部分混菌3种处理,经过5个月的盆栽试验表明,拮抗细菌HMB-1005能够在灭菌土和非灭菌土中定殖,且定殖数量达106cfu/g 土左右,并仍能保持较高的抑菌活性.通过对棉苗根内细菌的分离,证实拮抗细菌HMB-1005能在棉花根内定殖,数量达到103cfu/g.实验结果显

  3. 人工接种棉花黄萎病菌种子包衣效果研究%STUDIES ON ANTI- FUNGAL EFFECT OF SEED COATING FOR COTION BY ARTIFICIAL INOCUIATION OF VERTICILLIUM DAHLIAE KL E. B

    Institute of Scientific and Technical Information of China (English)

    廖中元; 柴勇; 何正明; 邓先明; 朱玉香; 秦森荣

    2001-01-01

    人工接种中等致病力的棉花黄萎病菌(VD),再用不同组分的种衣剂包衣处理种子,结果以20%克·福·多+0.1%消菌灵的效果最优,相对防效达65.6%,比标准对照药卫福40%200FF(相对防效为60.3%)增加5.3%,667m2产皮棉80.8kg,比对照增产34.7%,比卫福增产4.4%。%The spores of Verticillium dahliae Kleb with medium pathogenity were artificially inoculated onto the seeds of cotton before they were coated with different types of seed coating agents. The results showed that the anti- fungal effect of the seed coating agent was the best which contained 20% carbofuran- thiram- carbendazin plus 0.1 Xiao Jun Ling- a newly developed fungicide, its relative control effect being as high as 65.6%,or a 5.3% increase as compared with that of vitavax 200FF(60.3% )-a standard Ameriean seed coating agent. Lint cotton yield per mu was 80.8 kg,being 34.7% and4.4% higher than the blank control and the vitavax 200FF treatment.

  4. Fosfito de potássio na indução de resistência a Verticillium dahliae Kleb., em mudas de cacaueiro (Theobroma cacao L. Effect of potassium phosphite on the induction of resistance in cocoa seedlings (Theobroma cacao L. against Verticillium dahliae Kleb

    Directory of Open Access Journals (Sweden)

    Pedro Martins Ribeiro Júnior

    2006-08-01

    Full Text Available Há vários relatos da utilização de fosfitos no controle de doenças de plantas, por meio de ação direta, antifúngica e indireta por indução de resistência. Essa atuação como indutor é questionada, não sendo encontradas, em muitos desses trabalhos, evidências de respostas de defesa ativadas pelos sais de fosfito. Com o presente trabalho, objetivou-se estudar o efeito de doses (0,62; 1,25; 2,5 e 5 mL.L-1 de água de fosfito de potássio na indução de resistência em mudas de cacaueiro a V. dahliae, além de investigar os possíveis mecanismos envolvidos na resposta de defesa. Foram realizados experimentos no Laboratório de Fisiologia do Parasitismo e casa-de-vegetação do Departamento de Fitopatologia UFLA. A aplicação foliar do fosfito foi realizada 7 dias antes das inoculações e as avaliações de severidade foram realizadas aos 20, 30, 40, 50 e 60 dias após a inoculação. Foi realizado também um experimento para verificar o efeito tóxico direto e outro para avaliar a atividade das enzimas peroxidases e polifenoloxidases e a concentração de lignina. O tratamento com o fosfito de potássio (1,25 mL.L-1 de água proporcionou 10% de redução na área abaixo da curva de progresso da severidade da murcha-de-Verticillium (AACPD, 60 dias após aplicação foliar, não diferindo de nenhuma das doses, nem da testemunha. Todas as doses utilizadas apresentaram efeito fungitóxico, inibindo a germinação de V. dahliae. A aplicação do fosfito de potássio (1,25 mL.L-1 de água não induziu aumento na atividade das enzimas peroxidases e polifenoloxidases em relação à testemunha. Plantas tratadas com esse produto apresentaram um pequeno incremento na concentração de lignina, não apresentando diferença significativa da testemunha absoluta.There are several reports concerning with the use of phosphites for controlling of plant diseases, through direct action, antifungal, and indirect, by resistance induction. The

  5. Pseudomonas community structure and antagonistic potential in the rhizosphere : insights gained by combining phylogenetic and functional gene-based analyses

    NARCIS (Netherlands)

    Costa, Rodrigo; Gomes, Newton C. M.; Kroegerrecklenfort, Ellen; Opelt, Katja; Berg, Gabriele; Smalla, Kornelia

    2007-01-01

    The Pseudomonas community structure and antagonistic potential in the rhizospheres of strawberry and oilseed rape (host plants of the fungal phytopathogen Verticillium dahliae) were assessed. The use of a new PCR-DGGE system, designed to target Pseudomonas-specific gacA gene fragments in environment

  6. Improved genome sequence of the phytopathogenic fungus Rhizoctonia solani AG1-IB 7/3/14 as established by deep mate-pair sequencing on the MiSeq (Illumina) system.

    Science.gov (United States)

    Wibberg, Daniel; Rupp, Oliver; Jelonek, Lukas; Kröber, Magdalena; Verwaaijen, Bart; Blom, Jochen; Winkler, Anika; Goesmann, Alexander; Grosch, Rita; Pühler, Alfred; Schlüter, Andreas

    2015-06-10

    The phytopathogenic fungus Rhizoctonia solani AG1-IB of the phylum Basidiomycota affects various economically important crops comprising bean, rice, soybean, figs, cabbage and lettuce. The R. solani isolate 7/3/14 of the anastomosis group AG1-IB was deeply resequenced on the Illumina MiSeq system applying the mate-pair mode to improve its genome sequence. Assembly of obtained sequence reads significantly reduced the amount of scaffolds and improved the genome sequence of the isolate compared to the previous sequencing approach. The genome sequence of the AG1-IB isolate 7/3/14 now provides an up-graded basis to analyze genome features predicted to play a role in pathogenesis and for the development of strategies to antagonize the pathogenic impact of this fungus.

  7. The mononuclear nickel(II) complex bis(azido-κN)bis[2,5-bis(pyridin-2-yl)-1,3,4-thiadiazole-κ(2) N(2) ,N(3) ]nickel(II) protects tomato from Verticillium dahliae by inhibiting fungal growth and activating plant defences.

    Science.gov (United States)

    Zine, Hanane; Rifai, Lalla Aicha; Koussa, Tayeb; Bentiss, Fouad; Guesmi, Salaheddine; Laachir, Abdelhakim; Makroum, Kacem; Belfaiza, Malika; Faize, Mohamed

    2017-01-01

    The antifungal properties of the nickel(II) complex bis(azido-κN)bis[2,5-bis(pyridin-2-yl)-1,3,4-thiadiazole-κ(2) N(2) ,N(3) ]nickel(II) [NiL2 (N3 )2 ] and its parental ligand 2,5-bis(pyridin-2-yl)-1,3,4-thiadiazole were examined to evaluate their ability to protect tomato plants against Verticillium dahliae. Our main objectives were to determine their effects on the in vitro growth of the pathogen, and their aptitude for controlling verticillium wilt and activating plant defence responses in the greenhouse. NiL2 (N3 )2 exhibited in vitro an elevated inhibition of radial growth of three strains of the pathogen. According to the strain, the EC50 values ranged from 10 to 29 µg mL(-1) for NiL2 (N3 )2 . In the greenhouse, it induced an elevated protection against V. dahliae when it was applied twice as foliar sprays at 50 µg mL(-1) . It reduced the leaf alteration index by 85% and vessel browning by 96%. In addition, its protective ability was associated with the accumulation of H2 O2 and the activation of total phenolic content, as well as potentiation of the activity of peroxidase and polyphenol oxidase. These results demonstrated that the coordination of the ligand with Ni associated with the azide as a coligand resulted in an improvement in its biological activity by both inhibiting the growth of V. dahliae and activating plant defence responses. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  8. ダイズシストセンチュウに対するVerticillium lecanii プロトプラスト融合株の微生物防除資材としての可能性:プロトプラスト融合は植物寄生性線虫の生物的防除において有用な手法と成り得るか?

    OpenAIRE

    Shinya, Ryoji; Watanabe, Ai; Aiuchi, Daigo; Tani, Masayuki; KURAMOCHI, Katsuhisa; Kushida, Atsuhiko; KOIKE, MASANORI; 新屋, 良治; 渡部, 愛; 相内, 大吾; 谷, 昌幸; 倉持, 勝久; 串田, 篤彦; 小池, 正徳

    2008-01-01

    Verticillium lecanii is a potentially useful biological control agent (BCA) for soybean cyst nematode (SCN), Heterodera glycines. The objective of this study was to screen hybrid strains of V. lecanii, derived from protoplast fusion, for effective control agents of SCN, and to investigate whether the protoplast fusion technique is an effective tool for development of nematode control agents. Three parental strains (Vertalec®, Mycotal®, and B-2) and their 162 hybrid strains were screened in gr...

  9. Inibição do desenvolvimento de fungos fitopatogênicos por detergente derivado de óleo da mamona (Ricinus communis The castor oil plant detergent (Ricinus communis inhibits the asexual development of phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    Eunice Hitomi Takano

    2007-10-01

    Full Text Available No presente estudo avaliou-se o efeito fungitóxico do detergente derivado do óleo da mamona (Ricinus communis sobre o desenvolvimento dos fitopatógenos: Pyricularia grisea, Fusarium graminearum e Colletotrichum lindemuthianum. Seis concentrações do detergente (12,5mL L-1 a 300mL L-1 foram, individualmente, incorporadas ao Meio Basal; a seguir, após inoculação fúngica, o crescimento radial dos micélios foi avaliado. A inibição total do desenvolvimento de C. lindemuthianum e P. grisea foi observada entre as concentrações de 50mL L-1 e 200mL L-1, respectivamente. Com base no crescimento miceliano das colônias de F. graminearum, a atividade antifúngica do detergente do óleo da mamona (DOM determinou inibição variável entre 79,4 e 91% para a raça F2 e entre 80,7 e 90,7% para a raça F4. O detergente, nas concentrações de 100 a 300mL L-1, inibiu em 100% a germinação de conídios de F. graminearum (raças F-4 e F-2. Os resultados demonstram nítida atividade antifúngica do detergente derivado do óleo da mamona sobre fitopatógenos.In the present study the fungitoxic effect of the castor oil plant detergent (Ricinus communis on the development of the phytopathogens Pyricularia grisea, Fusarium graminearum and Colletotrichum lindemuthianum was evaluated. Six concentrations of the detergent (12.5mL L-1 to 300mL L-1 had been, individually, incorporated to the Basal Medium. After fungi inoculations, the radial growth of mycelia were evaluated. Detergent at 50mL L-1 and 200mL L-1 inhibited completely the development of P. grisea and C. lindemuthianum, respectively. On the basis of the mycelial growth of F. graminearum, the fungitoxic activity of the castor oil plant detergent (DOM determined inhibition in the range of 79.4 and 91% for the F2 race and 80.7 and 90.7% for the F4 race. Detergent at the concentrations of 100mL L-1 to 300mL L-1 inhibited in 100% the F. graminearum germination conidia (races F-4 and F-2. Results

  10. 环腺苷酸对向日葵大丽轮枝菌生物学特性及致病力的影响%Effect of cAMP on biological characteristics and pathogenicity of sunflower Verticillium dahliae

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    To unravel the effects of cAMP on biological characteristics and pathogenicity of sunflower pathogen Verticillium dahlia,growth rate,conidia production,germination rate of conidia spores,microsclerotia production, crude toxin and pathogenicity of V. dahliae were measured after V. dahlia cultured in medium with 10 mmol/L cAMP. Results showed that pathogen conidia was increased by 55. 45% in the medium,but conidia germination rate,microsclerotia yield,crude toxin production and disease index were reduced by 69%,46. 8%,35. 27% and 7. 28% respectively. In conclusion,exogenously cAMP promoted V. dahliae conidia yield,inhibited conidia germi-nation rate,and reduced microsclerotia yield,crude toxin production and pathogenicity. Among them,cAMP inhi-bition effect on microsclerotia formation was greater.%为探究环腺苷酸( cAMP,cyclic AMP)对向日葵大丽轮枝菌生物学特性及致病力的影响,在实验室条件下研究了外源添加cAMP(10mmol/L)后,向日葵大丽轮枝菌的菌落形态、生长速率、产孢量、分生孢子萌发率、微菌核数量、粗毒素分泌量以及致病力的变化。结果表明,外源cAMP使向日葵大丽轮枝菌分生孢子的产孢量提高了55.45%;而在含有cAMP的水琼胶培养基上培养24h后,大丽轮枝菌的分生孢子萌发率降低了69%;在含有cAMP的PDA上培养25d后,大丽轮枝菌微菌核形成的数量相比对照降低了46.8%;在含有cAMP的液体培养基中振荡培养15d后,大丽轮枝菌粗毒素的含量仅为0.156mg/mL,比对照培养液中毒素降低了35.27%。向日葵接种10mmol/L cAMP处理的大丽轮枝菌后,其病情指数为34.38,相比对照组降低了7.28%。因此,外源添加10mmol/L cAMP除了能够提高向日葵大丽轮枝菌的产孢量外,对其生长速率、分生孢子萌发率、微菌核形成的数量、粗毒素分泌量以及致病力均有不同程度的抑制作用,尤其是对微菌核

  11. Primary Analysis of Antifungal Proteins Produced by Bacillus amylolique faciens X-278 against Verticillium dahliae%解淀粉芽孢杆菌X-278菌株抗菌蛋白初步分析

    Institute of Scientific and Technical Information of China (English)

    黎波; 魏学军; 李亚宁; 孟庆芳; 张汀; 刘大群; 赤国彤

    2013-01-01

    为研究解淀粉芽孢杆菌X-278菌株所产抗菌蛋白的性质,通过饱和硫酸铵沉淀法提取X-278菌株蛋白粗提液,并对蛋白粗提液进行蛋白酶稳定性试验和热稳定性试验.结果表明,当硫酸铵饱和度为50%时,沉淀得到的X-278菌株蛋白粗提液对棉花黄萎病菌的押菌作用最强,抑菌圈直径迭18.9 mm.蛋白粗提液对蛋白酶K敏感性最差,对胰蛋白酶最敏感;蛋白粗提液经40℃处理后押菌活性最高,而121℃处理后抑菌活性最低,抑菌圈直径为13.6 mm,为对照(常温处理)的72%.以上结果表明,X-278菌株粗蛋白的部分抗菌成分属于耐热物质,并且抗菌成分间存在性质上的差异.%In order to study the characteristics of antifungal proteins isolated from Bacillus amyloliquefaciens X-278,crude proteins were extracted via precipitation of the cell culture of strain X-278 with ammonium sulphate,and the protease stability and thermal stability were determined.The result showed that the crude proteins of strain X-278 had strongest control effect on cotton Verticillium wilt using ammonium sulphate at 50% saturation,with the inhibition zone diameter of 18.9 mm.The crude protein extracts were relatively insensitive to proteinase K,but more sensitive to trypsin.The activity of the antifungal substrates was investigated under different temperatures,and the result showed that the antifungal activity at 40 ℃ was the highest,while it decreased most at 121 ℃ and showed significant difference from the control.However,the crude protein extracts still retained 72 % of the antifungal activity compared to the control and the diameter of inhibition zone was 13.6 mm after heat treatment for 30 min under 121 ℃.The results above indicated that part of the antifungal extracts of Bacillus amyloliquefaciens X-278 were heat-resistant substance and the antifungal extracts were several substrates with different traits.

  12. Improved DNA Extraction of Verticillium lecanii in Soil and Its Application%土壤蜡蚧轮枝菌DNA提取方法的改进及其应用效果

    Institute of Scientific and Technical Information of China (English)

    周洁; 张艳军; 谢明; 贾开志

    2014-01-01

    土壤真菌分子生态学研究需要大量和高效地提取真菌的 DNA。应用市售的一般试剂盒提取土壤虫生真菌DNA,常常存在得率和质量低的问题,甚至根本提取不到土壤真菌的DNA样品。针对这一问题,本研究对细胞破碎方式及后续DNA提取参数进行了一系列的改进和优化,建立了一套针对性的提取方法。该提取法对虫生真菌—蜡蚧轮枝菌纯培养物的灵敏度极高,可在10个孢子的条件下提取到DNA,而市售的试剂盒则不能提取到(试剂盒在107个孢子条件仍提不到DNA);该提取法对人工投菌土样的DNA提取灵敏度达到102个孢子/克土,所得的DNA纯度高,无明显抑制后续PCR扩增的物质;施用过蜡蚧轮枝菌的田间土样所得 DNA 样品可成功扩增出蜡蚧轮枝菌特异片段,而未施用过蜡蚧轮枝菌的田间土样所得DNA 样品不能扩增出蜡蚧轮枝菌特异片段。本提取方法具有灵敏度和纯度高的特点,适用于土壤虫生真菌的分子生态学样品制备。%It is required to efficiently extract large quantity and high quality of fungal DNA for the soil fungal molecular ecological study. However, the yield and quality of DNA are often insufficient or even no any product can be produced by the commercial DNA extraction kit. To solve these problems, the cell lysis and subsequent DNA extraction parameters were optimized in this study, and then an efficient method of extracting soil fungal DNA was established. The results showed this method was very sensitive to entomopathogenic fungi Verticillium lecanii. DNA could be extracted from the pure culture samples which contained more than 10 spores, and the artificial soil samples which contained more than 100 spores per gram soil. However commercial DNA extraction kits are unable to extract DNA from the pure culture samples which contained 107 spores. The DNA product from our method did not show any obvious inhibition in

  13. Complete inhibition of the tentoxin-resistant F1-ATPase from Escherichia coli by the phytopathogenic inhibitor tentoxin after substitution of critical residues in the alpha - and beta -subunit.

    Science.gov (United States)

    Schnick, Claudia; Körtgen, Nicole; Groth, Georg

    2002-12-27

    Substitution of critical residues in the alpha- and beta-subunit can turn the typically resistant ATP synthase from the bacterium Escherichia coli into an enzyme showing high sensitivity to the phytopathogenic inhibitor tentoxin, which usually affects only certain sensitive plant species. In contrast to recent results obtained with the thermophilic F(1) (Groth, G., Hisabori, T., Lill, H., and Bald, D. (2002) J. Biol. Chem. 277, 20117-20119), substitution of a critical serine in the beta-subunit (betaSer(59)), which is supposed to provide an important intermolecular hydrogen bond in the binding site, was not sufficient on its own for conferring tentoxin sensitivity to the E. coli F(1) complex. Superimposition of the chloroplast F(1)-tentoxin inhibitor complex on a homology model of the E. coli F(1) complex provided detailed information on the critical residues in the alpha-subunit of the binding cleft and allowed us to model the binding site according to the steric requirements of the inhibitor. Substitution of the highly conserved residue alphaLeu(64) seems to be most important for allowing access of the inhibitor to the binding site. Combining this substitution with either additional replacements in the alpha-subunit (Q49A, L95A, E96Q, I273M) or the replacement of Ser(59) in the beta-subunit enhanced the sensitivity to the inhibitor and resulted in a complete inhibition of the E. coli F(1)-ATPase by the plant-specific inhibitor tentoxin.

  14. A multi-repeat adhesin of the phytopathogen, Pectobacterium atrosepticum, is secreted by a Type I pathway and is subject to complex regulation involving a non-canonical diguanylate cyclase.

    Science.gov (United States)

    Pérez-Mendoza, Daniel; Coulthurst, Sarah J; Humphris, Sonia; Campbell, Emma; Welch, Martin; Toth, Ian K; Salmond, George P C

    2011-11-01

    Cyclic diguanylate (c-di-GMP) is a second messenger controlling many important bacterial processes. The phytopathogen Pectobacterium atrosepticum SCRI1043 (Pba1043) possesses a Type I secretion system (T1SS) essential for the secretion of a proteinaceous multi-repeat adhesin (MRP) required for binding to the host plant. The genes encoding the MRP and the T1SS are tightly linked to genes encoding several putative c-di-GMP regulatory components. We show that c-di-GMP regulates secreted MRP levels in Pba1043 through the action of two genes encoding predicted diguanylate cyclase (DGC) and phosphodiesterase proteins (ECA3270 and ECA3271). Phenotypic analyses and quantification of c-di-GMP levels demonstrated that ECA3270 and ECA3271 regulate secreted MRP levels by increasing and decreasing, respectively, the intracellular levels of c-di-GMP. Moreover, ECA3270 represents the first active DGC reported to have an alternative active-site motif from the 'canonical' GG[D/E]EF. ECA3270 has an A-site motif of SGDEF and analysis of single amino acid replacements demonstrated that the first position of this motif can tolerate functional substitution. Serine in position one of the A-site is also observed in many other DGCs. Finally, another T1SS-linked regulator (ECA3265) also plays an important role in regulating secreted MRP, with an altered localization of MRP observed in an ECA3265 mutant background. Mutants defective in these three T1SS-linked regulators exhibit a reduction in root binding and virulence, confirming that this complex, finely tuned regulation system is crucial in the interaction with host plants.

  15. Chemical Composition and Anti -fungal Properties of Xinjiang Propolis Extracts against Agricultural Phytopathogenic Fungi%新疆产蜂胶提取物对农作物病原真菌的抗性作用

    Institute of Scientific and Technical Information of China (English)

    阿米尼姑丽·买买提; 木塔力甫·艾买提; 尼砸木·艾海提; 布威海丽且姆·阿巴拜科日; 依米提·热合曼

    2011-01-01

    [目的]分析新疆蜂胶的化学成分并研究其抗菌活性.[方法]用超声波提取法提取蜂胶,采用GC-MS联用技术鉴定其化学成分,通过最小抑菌浓度试验和抑菌试验来测定蜂胶的抗真菌活性.[结果]从蜂胶的乙醇提取物中鉴定出23种化合物.蜂胶对不同真菌表现出不同的抗菌活性,对不同植物病原真菌的抑菌率为16.7%~39.6%,最小抑菌浓度为2.0~4.0 mg/mL.[结论]新疆蜂胶具有广谱的抑菌活性,对某些重要植物病原菌有一定防控作用.%[Objective] The purpose of this project was to analyze the chemical composition of Xinjiang propolis extracts and investigate its antifungi activity. [ Method ] Ultrasonic was used to extract propolis and Gas Chromatography - Mass Spectrometry were applied to identify its chemical composition. Through the tests of minimum inhibitory concentrations (MIC) and mycelial growth inhabitation, its antifungal activity assays were determined. [ Result] twenty - three compounds were identified in ethanol - extracted propolis (EEP). Propolis displayed different antifungi actions depending on the type of fungi tested. It showed the antifungi activity range was from 16.7% to 39.6% and minimum inhibitory concentration range was from 2.0 to 4. Omg/ mL against the tested phytopathogenic fungi as a mycelial growth inhibitor. [ Conclusion ] The results obtained in this study demonstrated that Xinjiang propolis extracts possessed a wide range spectrum of fungistatic activity and could become an alternative to controlling certain important plant fungal diseases.

  16. Antifungal Activity of Five Solanaceous Glycoalkaloids and Their Mixtures against Phytopathogenic Fungi Cercosporella brassicae and Alternaria porri%五种茄科糖苷生物碱及其混合物的抗真菌活性研究

    Institute of Scientific and Technical Information of China (English)

    赵雪淞; 高聆; 王娟; 徐文静; 周义发

    2009-01-01

    The antifungal activity of five solanaceae glycoalkaloids solanine,chaconine,solasonine,solamargine and tomatine against phytopathogenic fungi Cercosporella brassicae and Alternaria porri has been evaluated.Tomatine showed the highest antifungal activity against C.brassicae and A.porri among five compounds,followed with chaconie,solamargine and solasonine,while solanine showed the lowest antifungal activity.Mixture of potato glycoalkaloids solanine and chaconine produced marked synergistic antifungal activity.The magnitude of synergisms is higher at lower concentrations than that at higher concentrations.The antifungal activity of individual glycoalkaloid against A.porri was relative low,even no activity,but the mixture of chaconine and solanine showed significant synergism.There was no synergism between glycoalkaloids solasonine and solamargine from Solanum nigrum in inhibiting fungul growth.The mixtures of solamargine and chaconine,and solasonine and solanine both caused additive inhibition on the growth of fungi.%本文研究了五种茄科糖苷生物碱(茄碱、查茄碱、边缘茄碱、澳洲茄碱和番茄碱)对两种植物病原真菌白菜白斑病菌和葱紫斑病菌的抑制活性.结果表明番茄碱的抗真菌活性最强,其后依次是查茄碱、边缘茄碱和澳洲茄碱,茄碱的活性最弱;不同浓度茄碱和查茄碱(马铃薯中的两种糖苷生物碱)的混合物均具有协同抗真菌作用,且低浓度的混合物产生的协同作用效果较大;边缘茄碱和澳洲茄碱(龙葵中的两种糖苷生物碱)的混合物基本没有协同抗真菌作用;边缘茄碱和查茄碱的混合物以及澳洲茄碱和茄碱的混合物(均为来自不同植物的糖苷生物碱的混合物)在抗真菌活性上都呈现了相加关系.

  17. Antibiotic multiresistance plasmid pRSB101 isolated from a wastewater treatment plant is related to plasmids residing in phytopathogenic bacteria and carries eight different resistance determinants including a multidrug transport system.

    Science.gov (United States)

    Szczepanowski, Rafael; Krahn, Irene; Linke, Burkhard; Goesmann, Alexander; Pühler, Alfred; Schlüter, Andreas

    2004-11-01

    Ten different antibiotic resistance plasmids conferring high-level erythromycin resistance were isolated from an activated sludge bacterial community of a wastewater treatment plant by applying a transformation-based approach. One of these plasmids, designated pRSB101, mediates resistance to tetracycline, erythromycin, roxythromycin, sulfonamides, cephalosporins, spectinomycin, streptomycin, trimethoprim, nalidixic acid and low concentrations of norfloxacin. Plasmid pRSB101 was completely sequenced and annotated. Its size is 47 829 bp. Conserved synteny exists between the pRSB101 replication/partition (rep/par) module and the pXAC33-replicon from the phytopathogen Xanthomonas axonopodis pv. citri. The second pRSB101 backbone module encodes a three-Mob-protein type mobilization (mob) system with homology to that of IncQ-like plasmids. Plasmid pRSB101 is mobilizable with the help of the IncP-1alpha plasmid RP4 providing transfer functions in trans. A 20 kb resistance region on pRSB101 is located within an integron-containing Tn402-like transposon. The variable region of the class 1 integron carries the genes dhfr1 for a dihydrofolate reductase, aadA2 for a spectinomycin/streptomycin adenylyltransferase and bla(TLA-2) for a so far unknown Ambler class A extended spectrum beta-lactamase. The integron-specific 3'-segment (qacEDelta1-sul1-orf5Delta) is connected to a macrolide resistance operon consisting of the genes mph(A) (macrolide 2'-phosphotransferase I), mrx (hydrophobic protein of unknown function) and mphR(A) (regulatory protein). Finally, a putative mobile element with the tetracycline resistance genes tetA (tetracycline efflux pump) and tetR was identified upstream of the Tn402-specific transposase gene tniA. The second 'genetic load' region on pRSB101 harbours four distinct mobile genetic elements, another integron belonging to a new class and footprints of two more transposable elements. A tripartite multidrug (MDR) transporter consisting of an ATP

  18. Conserved signature indels and signature proteins as novel tools for understanding microbial phylogeny and systematics: identification of molecular signatures that are specific for the phytopathogenic genera Dickeya, Pectobacterium and Brenneria.

    Science.gov (United States)

    Naushad, Hafiz Sohail; Lee, Brian; Gupta, Radhey S

    2014-02-01

    Genome sequences are enabling applications of different approaches to more clearly understand microbial phylogeny and systematics. Two of these approaches involve identification of conserved signature indels (CSIs) and conserved signature proteins (CSPs) that are specific for different lineages. These molecular markers provide novel and more definitive means for demarcation of prokaryotic taxa and for identification of species from these groups. Genome sequences are also enabling determination of phylogenetic relationships among species based upon sequences for multiple proteins. In this work, we have used all of these approaches for studying the phytopathogenic bacteria belonging to the genera Dickeya, Pectobacterium and Brenneria. Members of these genera, which cause numerous diseases in important food crops and ornamental plants, are presently distinguished mainly on the basis of their branching in phylogenetic trees. No biochemical or molecular characteristic is known that is uniquely shared by species from these genera. Hence, detailed studies using the above approaches were carried out on proteins from the genomes of these bacteria to identify molecular markers that are specific for them. In phylogenetic trees based upon concatenated sequences for 23 conserved proteins, members of the genera Dickeya, Pectobacterium and Brenneria formed a strongly supported clade within the other Enterobacteriales. Comparative analysis of protein sequences from the Dickeya, Pectobacterium and Brenneria genomes has identified 10 CSIs and five CSPs that are either uniquely or largely found in all genome-sequenced species from these genera, but not present in any other bacteria in the database. In addition, our analyses have identified 10 CSIs and 17 CSPs that are specifically present in either all or most sequenced Dickeya species/strains, and six CSIs and 19 CSPs that are uniquely found in the sequenced Pectobacterium genomes. Finally, our analysis also identified three CSIs

  19. Expression Differences of Dirigent-Like Protein Genes in Upland Cotton Responsed to Infection by Verticillium dahliae%黄萎病菌侵染下陆地棉Dirigent-like蛋白基因表达差异分析

    Institute of Scientific and Technical Information of China (English)

    郭宝生; 师恭曜; 王凯辉; 刘素恩; 赵存鹏; 王兆晓; 耿军义; 华金平

    2014-01-01

    [Objective]The objective of this study is to explore the Dirigent-like protein gene family and to figure out polymorphism of genes and expression differences between resistant and susceptible varieties of Upland cotton (Gossypium hirsutum) in responding to infection by Verticillium dahliae.[Method]Dirigent-like protein genes were acquired using Illumina Hiseq 2000 RNA-Seq transcriptional analysis. Multiple sequence alignment of encoding proteins was carried using online sequence analysis softwares and bioinformatics analysis tools for phylogenetic analysis. Quantitative real-time PCR with gene-specific primers was employed to identify the expression differences between resistant variety‘Ji 79’ and susceptible variety‘TM-1’, at 0, 1, 8, 24, 48, 72 h, and 96 h after inoculation. The quantitative reactions were performed on the ABI 7500 Real-Time PCR System with 7500 System Software (Applied Biosystems, USA) using the SYBR Green Master Mix. The comparative 2-ΔΔCt method of quantification was used with cotton GhACTIN as the reference.[Result]Totally 12 members of this gene family were found in Gossypium hirsutum, and phylogenetic analysis demonstrated 3 distinct subfamilies and 4 relatively conservative domains existed. The members of Dirigent-like protein genes were located on chromosomes 2, 3, 6, 7, 9, 10 and 13 in Gossypium raimondii, and some located in clusters on the same chromosome of D5 genome. Differential expression analysis indicated that dynamic response existed in different varieties. There were 6 members (GhDIR4, GhDIR6, GhDIR7, GhDIR9, GhDIR10 and GhDIR11) up-regulated (RQ≥2) in resistant variety Ji 79 after 1h inoculation, and RQ of GhDIR9 reached about 6.5;and 4 members (GhDIR1, GhDIR4, GhDIR7 and GhDIR11) up-regulated (RQ≥1) after 8h inoculation, only GhDIR4 up-regulated (RQ≥2) after 24h inoculation, GhDIR1, GhDIR4, GhDIR7 and GhDIR11 up-regulated (RQ≥2) after 48h inoculation;only GhDIR9 up-regulated (RQ=6.4) after 72h inoculation

  20. The synergetic inhibition effect of arbuscular mycorrhizal fungi cooperated Bacillus vallismortis organic fertilizer on cotton Verticillium wilt%菌根真菌和死谷芽孢杆菌生物有机肥对连作棉花黄萎病的协同抑制

    Institute of Scientific and Technical Information of China (English)

    张国漪; 丁传雨; 任丽轩; 沈其荣; 冉炜

    2012-01-01

    通过温室盆栽试验,育苗期接种丛枝菌根真菌(AMF),移栽时接种死谷芽孢杆菌(Bacillus vallismortis HJ-5)、有机肥(OF)或者生物有机肥(HJ-5OF),研究了不同处理对棉花黄萎病和棉花生长的影响.结果表明:配合施用AMF与HJ-5OF显著降低了棉花黄萎病的发病指数,与对照相比其病情指数下降了72.80%;根际可培养微生物数量和种类发生显著变化,施用AMF与HJ-5OF处理的真菌、大丽轮枝菌菌核数量分别比对照下降了68.88%、47.20%,细菌、放线菌数量分别为对照的23.81倍、2.40倍;根际区系中,真菌的种类显著降低,细菌的种类显著增加;AMF和HJ-5OF的协同作用,显著提高了棉花的生物量,促进了棉花对磷素的吸收.结论:接种AMF提高HJ-5OF对棉花黄萎病的防治效果,促进棉花的生长,改善土壤微生物区系.%Pot experiment was carried out to investigate the effect of arbuscular mycorrhizal fungus(AMF) and Bacillus vallismortis HJ-5 on cotton growth and Verticillium wilt by inoculation of AMF on seedling nursery stage and bio-organic fertilizer after transplanting. The results showed that the disease index significantly decreased by 72. 80% as compared with the non-inoculation control unde combined application of AMF and bio-organic fertilizer. The microbial communities were improved by inoculation of bio-agents. The number of fungi and sclerotia decreased by 68. 88% and 47. 20% compared with control, respectively; in addition, the number of bacteria and actinomyces increased by 23.81 times and 2.40 times compared with control,respectively. The fungi species significantly reduced and the bacteria species reduced in the rhizosphere. The number of cottons biomass and phosphorus absorption was significantly promoted by inoculation of AMF and HJ-50F. In conclusion,HJ-5OF bio-organic fertilizer associated with AMF significantly decreased the Verticillium wilt,promoted cotton growth,and altered the soil microbial

  1. Phytopathogen-induced changes to plant methylomes.

    Science.gov (United States)

    Hewezi, Tarek; Pantalone, Vince; Bennett, Morgan; Neal Stewart, C; Burch-Smith, Tessa M

    2017-07-29

    DNA methylation is a dynamic and reversible type of epigenetic mark that contributes to cellular physiology by affecting transcription activity, transposon mobility and genome stability. When plants are infected with pathogens, plant DNA methylation patterns can change, indicating an epigenetic interplay between plant host and pathogen. In most cases methylation can change susceptibility. While DNA hypomethylation appears to be a common phenomenon during the susceptible interaction, the levels and patterns of hypomethylation in transposable elements and genic regions may mediate distinct responses against various plant pathogens. The effect of DNA methylation on the plant immune response and other cellular activities and molecular functions is established by localized differential DNA methylation via cis-regulatory mechanisms as well as through trans-acting mechanisms. Understanding the epigenetic differences that control the phenotypic variations between susceptible and resistant interactions should facilitate the identification of new sources of resistance mediated by epigenetic mechanisms, which can be exploited to endow pathogen resistance to crops.

  2. Controle de fitopatógenos do solo com materiais vegetais associados à solarização Control of soil-borne phytopathogenic fungi by the association between materials vegetable materials and solarization

    Directory of Open Access Journals (Sweden)

    Márcia Michelle de Queiroz Ambrósio

    2008-12-01

    Full Text Available A incorporação de material orgânico associada à solarização do solo é uma técnica promissora no controle de patógenos de plantas. O trabalho consistiu na prospecção de materiais vegetais promissores na produção de voláteis fungitóxicos capazes de inviabilizar as estruturas de resistência de fitopatógenos do solo. Em condição de campo foram incorporados 3 Kg/m² de folhas e ramos de brócolos, eucalipto, mamona e mandioca brava, associada ou não à solarização, visando o controle de Fusarium oxysporum f. sp. lycopersici raça 2; Macrophomina phaseolina; Rhizoctonia solani AG-4 HGI e Sclerotium rolfsii. O controle foi avaliado por meio da sobrevivência das estruturas, em meios semi-seletivo específicos, aos 7, 14, 21 e 28 dias do início do experimento. Foram monitoradas as temperaturas do solo e do ar por um DataLogger Tipo CR23X (Campbell Scientific e a porcentagem de CO2 e de O2 pelo equipamento analisador de gases (Testo 325-1. A associação da incorporação dos materiais vegetais com a solarização do solo inativou F. oxysporum f. sp. lycopersici raça 2, M. phaseolina e R. solani. O fungo S. rolfsii foi o único que não apresentou 100% de controle com solarização mais mamona durante o período estudado. A incorporação de mandioca seguido de solarização propiciou o controle de todos os fungos estudados com menos de sete dias da instalação do experimento, sendo tão eficiente quanto o brócolos na erradicação dos fitopatógenos veiculados pelo sol.The association between previous incorporation of vegetable material and soil solarization is a promising technique for control of several phytopathogens. The objective of this work consisted in the investigation of materials vegetable that are promising to produce fungitoxic volatiles capable of inactivating the resistence structures of soil plant pathogens. Three Kg/m² of vegetable materials were incorporated under field conditions, and the survival of four

  3. Antifungal Activity of the Ozonized Sunflower Oil (Oleozon) to the Phytopathogenic Fungus%臭氧化葵花油对植物病原真菌的抗性研究

    Institute of Scientific and Technical Information of China (English)

    李勃; 柯杨; 马瑜; 陈志杰

    2013-01-01

    Effects of ozonized sunflower oil (oleozon) on Penicillium expansum, Botrytis cinerea, Fusarium oxysporum, Alternaria solani and Cytospora mandshurica was evaluated. Minimum inhibitory concentrations (MICs) determined with agar dilution method indicated that oleozon was a valuable antimicrobial compound against phytopathogenic fungus, with an MIC ranging from 0.86 to 2.48 mg·mL−1. C. mandshurica, P. expansum and B. cinerea were more susceptible to oleozon than other species tested. Oleozon reduced DNA and RNA contents to 0.08 and 0.21 mg·g−1 dry weight of hyphae in C. mandshurica, respectively. Oleozon significantly suppressed activities of extracellular amylase, lipase and casease in all tested fungal species. These results proved oleozon was a broad spectrum antimicrobial agent.%  通过利用臭氧化葵花油(Oleozon)对拓展青霉 Penicillium expansum、灰葡萄孢霉 Botrytis cinerea、尖孢镰刀菌 Fusarium oxysporum、茄链格孢霉 Alternaria solani 及苹果壳囊孢 Cytospora mandshurica 的抗菌试验来评价其对不同植物病原真菌的抗性。采用平板稀释法进行的最小抑制浓度试验(MICs)证明, Oleozon 对于所有供试菌株皆具有显著抗性,其 MIC 值范围在0.86~2.48 mg·mL−1之间。其中,苹果壳囊孢,拓展青霉及灰葡萄孢霉对于 Oleozon 较其他供试菌株更为敏感。苹果壳囊孢经 Oleozon 处理后,其菌体增长量最低,仅为0.09 g·100 mL−1。相应地,各供试菌在生长过程中细胞的核酸含量也大大降低,苹果壳囊孢单位菌体干重中的 DNA 及 RNA 含量分别仅为0.08和0.21 mg·g−1。同时,各处理组中病原菌的胞外淀粉酶、脂肪酶及酪蛋白酶的活力也同样受到明显抑制。本试验证明,Oleozon 对于不同类型的植物病原真菌具有广谱抗性。

  4. Synthesis and screening for potential against phytopathogenic fungi activity of novel amides%新型酰胺类化合物的合成及抗植物病原真菌活性研究

    Institute of Scientific and Technical Information of China (English)

    周国萍; 刘伟; 金洪; 陶科; 侯太平

    2012-01-01

    In order to search for novel agrochemicals with potential anti-phythopathogenic fungi activity, a series of daphneone analogues were designed and synthesized. They were screened for antifungal activities against five phytopathogenic fungi; Rhizoctonia solani, Botrytis cirerea , Gibberella zeae , Bipolaris maydis, and Sclerotia sclerotium. The preliminary bioassays indicated that some compounds exhibited a fairly good activity. N-(2-fluorophenyl)-2, 4, 5-trimethyl-3-furancarboxamide (p) showed a strong fungistatic activity against R. solani (98% and 99% growth inhibition at 20 and 200 mg/L, respectively). Two compounds, N-(4-fluorophenyl)-2, 5-dimethyl-3-furancarboxamide (h) and N-(2-fluorophenyl)-2, 5-dimethyl-3-furancarboxamide (k) at 200 mg/L inhibited the growth of Sclerotia sclerotium at 94% and 90% , respectively. The EC50 values for compound k were 0. 034 mg/L, while for the control fungicide carbendazim were 0. 050 mg/L. According to the EC50 and the preventive activity of compound k, it can be inferred that compound k had a very good activity against Rhizoctonia solani. Thus, the compound k -was demonstrated to be the most promising candidate for further study.%为了寻找具有潜在抗真菌活性的新型农用化学品,我们以瑞香狼毒中提取的二苯酮类似物为先导,设计并合成了一系列的酰胺类化合物,并进行了以下5种植物真菌的筛选:水稻纹枯病菌(Rhizoctonia solani)、小麦赤霉病菌(Gibberella zeae)、玉米小斑病菌(Bipolaris maydis)、番茄灰霉病菌(Botrytis cirerea)和油菜菌核病菌(Sclerotinia sclerotiorum).初步的活性筛选研究显示:氮-(2-氟苯基)-2,4,5-三甲基-3-呋喃甲酰胺(p)具有很强的抗水稻纹枯病菌活性(在20和200 mg/L的浓度下抑制率分别为98%和99%);氮-(4-氟苯基)-2,5-二甲基-3-呋喃甲酰胺(h)和氮-(2-氟苯基)-2,5-二甲基-3-呋喃甲酰胺(k)这两种化合物在200 mg/L浓度下对油菜菌核病菌的抑制率分别为94%和90

  5. Antifungal activity of medicinal plant extracts against phytopathogenic fungus Alternaria spp Actividad antifúngica de extractos de plantas medicinales contra el hongo fitopatógeno Alternaria spp

    Directory of Open Access Journals (Sweden)

    Paola Díaz Dellavalle

    2011-06-01

    Full Text Available The aim of the study was to evaluate the antifungal activity of extracts of 10 plant species used in traditional Uruguayan medicine against the phytopathogenic fungus Alternaria spp. The plants were selected on the basis of their reported ethnobotanical uses. Aqueous, saline buffer and acid extracts of different plant species were screened in vitro for their antifungal activity against Alternaria spp. For the antifungal evaluation we used a microspectrophotometric assay. Minimal inhibitory concentration (MIC and minimum fungicidal concentration (MFC of the extracts were determined. Three solvents were assayed on different tissues of the plants and among the 29 evaluated extracts, 31% of the extracts inhibited growth, similar to the effects of a chemical fungicide. Acid extracts of the plants were more effective than the aqueous or buffer extracts against Alternaria spp. The MIC values of the extracts were determined ranging between 1.25 and 25 µg mL-1. The MFC values of the extracts ranged between 1.25 µg mL-1 (Rosmarinus officinalis L. and 10 µg mL-1 (Cynara scolymus L.. MICs and MFCs values obtained from leaves (Salvia officinalis L. and R. officinalis and seeds extracts (Salvia sclarea L. were quite comparable to values obtained with the conventional fungicide captan (2.5 µg mL-1. The extracts of Salvia sclarea, S. officinalis and R. officinalis could be considered as potential sources of antifungal compounds for treating diseases in plants. These extracts showed maximum activity, even at very low concentrations, and the same fungicide effects as chemical fungicide. We conclude from this that these extracts exhibit amazing fungicidal properties that support their traditional use as antiseptics.El objetivo de este trabajo fue evaluar la actividad antifúngica de extractos vegetales de 10 especies utilizadas en la medicina tradicional uruguaya contra el hongo fitopatógeno Alternaria spp. Las plantas fueron seleccionadas en base a usos

  6. Establishment of a screening system for Tn5 insertion mutants of Bacillus pumilus DX01 with different anti-phytopathogenic activities%短小芽胞杆菌DX01菌株Tn5转座突变株的抑菌活性筛选体系的建立

    Institute of Scientific and Technical Information of China (English)

    胡晓璐; 陈云鹏; 沈新迁; 刘通; 顾振芳

    2012-01-01

    为建立短小芽胞杆菌Bacillus pumilus DX01菌株的Tn5转座突变株抑菌活性的高效筛选体系,采用发酵液平板抑菌法研究了各突变株发酵液对稻瘟病菌Magnaporthe grisea生长的影响,并测定了目标突变株的几丁质酶和蛋白酶活性及发酵液对稻瘟病菌分生孢子萌发的抑制率。从2 633个突变株中筛选出6个抑菌活性较对照菌株DX01显著变化的突变株。对照DX01发酵液对真菌孢子萌发的抑制率为36%,而突变株Tn5-901和Tn5-194则分别为96%和3%,抑菌活性与对照的差异达到极显著水平。其余4个突变株的几丁质酶、蛋白酶活性多重比较结果与发酵液平板抑菌结果并不完全一致,但发酵液平板抑菌法简单、高效,适用于短小芽胞杆菌突变株的抑菌活性初筛。%Inhibition of mycelial growth and conidial germination of the phytopathogenic fungus, Magna- porthe grisea by zymotic fluids of Bacillus pumilus mutants and enzymatic activities of chintinase and pro- teinase of the identified mutants were analyzed in order to establish an efficient screening system for an- tagonistic abilities in Tn5 transposon library of the B. pumilus strain DX01 and further to clone genes asso- ciated with anti-phytopathogenic activity. A total of 2 633 mutants were subjected to anti-phytopathogenic activity screening and six mutants were preliminarily identified. The tests revealed that the inhibition per- centage of M. grisea conidia that respectively treated with zymotic fluids of mutants TnS-901 and Tn5-194 and the wild-type strain DX01 were 96% , 3% , and 36%. A statistically significant difference in anti- bacterial activity between the two mutants and the control strain DX01 were found, but the other four mu- tants did not exhibit a unanimous result in the multiple comparison tests for enzymatic activities and inhi- bition of mycelial growth with bacterial zymotic fluids. In conclusion, the evaluation of inhibition of fun- gal mycelial

  7. Efeito "in vitro" de antibióticos e rizobactérias no controle de bactérias fitopatogênicas ao Eucalyptus spp. "In vitro" effect of antibiotics and rhizobacteria on the control of phytopathogenic bacteria in Eucalyptus spp.

    Directory of Open Access Journals (Sweden)

    Jeane de Fátima Cunha

    2006-12-01

    Full Text Available Doenças causadas por bactérias constituem um novo desafio à cultura do Eucalyptus spp., podendo, inclusive, limitar o uso de clones suscetíveis. O presente trabalho objetivou avaliar a eficiência de antibióticos e rizobactérias na inibição do crescimento "in vitro" de isolados de bactérias fitopatogênicas ao Eucalyptus spp. na fase de viveiro e de campo. O antibiótico sulfato de amicacina e a rizobactéria S1 (Bacillus subtillis destacaram-se quanto à inibição do crescimento do isolado fitopatogênico IP1-05 (Pseudomonas chichorii, enquanto a cefoxitina causou maior inibição dos isolados BSV16 e RVV11 (Rhizobium sp.. Os antibióticos de uso comercial na área agronômica, Mycoshield (oxitetraciclina e Agrimicina (estreptomicina e tetraciclina foram pouco efetivos. Este trabalho proporciona embasamento a alternativas para controle biológico de doenças bacterianas em mudas de Eucalyptus spp. na fase de viveiro.Diseases caused by bacteria represent a new challenge for Eucalyptus spp. and may also limit the use of susceptible clones. Our study aimed at an evaluation of the efficiency of antibiotics and rhizobacteria in inhibiting "in vitro" growth of phytopathogenic bacteria isolates in Eucalyptus spp. during the nursery stage and in the field. The antibiotic amicacine sulfate and rhizobacterium S1 (Bacillus subtilis stood out inhibiting the growth of the phytopathogenic isolate IP1-05 (Pseudomonas chichorii, whereas cefoxitin caused a greater inhibition of the isolates BSV16 and RVV11 (Rhizobium sp.. The commercial antibiotics for agricultural use Mycoshield (oxitetracycline and Agrimycin (estreptomycin and tetracycline were little effective. This study offers a base for alternatives of biological control of bacterial diseases in Eucalyptus spp. nursery seedlings.

  8. 利用寄主诱导的基因沉默技术验证大丽轮枝菌糖代谢相关基因的致病力%Verification of Verticillium dahliae Pathogenicity of Glycometabolism Related Genes by Using Host-Induced Gene Silencing Method

    Institute of Scientific and Technical Information of China (English)

    赵玉兰; 苏晓峰; 程红梅

    2015-01-01

    [Objective] The objective of this study is to screen glycometabolism related genes from Verticillium dahliae, confirm the relationship between those genes and the growth and pathogenicity of V. dahliae, and to provide a theoretical basis for the control of plant Verticillium wilt. [Method] The glycometabolism related genes of V. dahliae were screened using host-induced gene silencing (HIGS) method. The Tobacco rattle virus vectors containing target genes were injected into tobacco for transient expression of those genes, then V.d991 was inoculated to those transgenic tobacco using the root-dip approach, PDS would be whitened 7-10 days later, and their disease indexes were counted after 14 days. The fluorescence quantitative PCR was used to detect the amount of V.d991 biomass and the expression of VDPDF-1, VDEG-1, VDMAN-1and VDPD-1. [Result] Four glycometabolism related genes from V. dahliae (VDPDF-1, VDEG-1, VDMAN-1 and VDPD-1) were selected because of the great change of their disease indexes. Transgenic tobacco disease indexes of these four genes were reduced by 35%, 30%, 20%, and 10%, respectively, compared with the negative control. After injected with the mixture of VDEG-1/VDPDF-1, VDPD-1/VDPDF-1/VDMAN-1, VDPD-1/VDMAN-1, VDPD-1/VDPDF-1, and VDEG-1/VDPD-1, transgenic tobacco disease indexes were reduced by 45%, 45%, 40%, 40%, and 35%. In addition to VDPD-1 transgenic tobacco, the transgenic tobacco disease indexes had significant differences compared with the control. After inoculation with V.d991, the V.d991 biomass and expression of target gene were decreased, mixed gene injection had better interference effect than the single gene. The most obvious biomass reduction were EG/PDF (VDEG-1/VDPDF-1) and EG/PD (VDEG-1/VDPD-1), reached to 0.94, and the most significant interference effect was VDPDF-1, from PD/PDF (VDPD-1/VDPDF-1),reduced by 0.91.[Conclusion]Four genes were selected from V. dahliae by HIGS method. When these genes were interfered, those plants had

  9. Screening and identification of antagonistic spore bacterium S12 against cotton Verticillium wilt and preliminary study on its antagonistic mechanisms%棉花黄萎病拮抗芽孢杆菌S12的筛选鉴定及拮抗机制的分析

    Institute of Scientific and Technical Information of China (English)

    李全胜; 谢宗铭; 张国丽; 武冬梅; 马盼盼

    2015-01-01

    [目的]近年来,新疆棉田由于长期连作导致棉花黄萎病呈逐年加重的趋势,筛选适合新疆特殊地理环境的高效拮抗菌株对防治棉花黄萎病尤为重要.[方法]采用平板对峙和琼脂平板扩散法从生长健康的棉花根际土壤中筛选拮抗细菌,利用常规PCR方法扩增拮抗细菌16S rRNA基因序列,应用BLAST进行同源性搜索,通过构建系统发育树并结合生理生化检测对拮抗细菌进行鉴定.用底物水解法检测其产生水解酶的能力,PCR方法检测其产生抗菌肽的潜力.[结果]枯草芽孢杆菌S12对大丽轮枝菌(Verticillium dahliae Kleb.)的抑菌率为56%,抑菌圈直径为20 mm,其发酵液对病原菌孢子萌发抑制率为95.93%,表现出较好的抑菌能力;菌株S12能合成蛋白酶,含有srfAA、spaS、bacA抗菌肽合成基因,具有一定的生防潜力.[结论]获得了可抑制大丽轮枝菌的芽孢杆菌菌株S12,并分析了其拮抗机制,为新疆棉花黄萎病的生物防治提供菌株资源和理论指导.

  10. 植物病原真菌对几类重要单位点杀菌剂的抗药性分子机制%Molecular Basis of Resistance of Phytopathogenic Fungi to Several Site-Specific Fungicides

    Institute of Scientific and Technical Information of China (English)

    詹家绥; 吴娥娇; 刘西莉; 陈凤平

    2014-01-01

    Site-specific fungicides play an important role in plant disease management. However, frequent applications of the fungicides over a large geographic scale can induce the emergence of resistant strains in the pathogen population. Resistance to fungicides with various modes of action has been documented in many plant fungal pathogens. This review summaries the current advances in understanding of the modes of action in five major classes of site-specific fungicides including methyl benzimidazole carbamate (MBCs), dicarboximide fungicides (DCFs), 14α-demethylase inhibitors (DMIs), quinone outside inhibitors (QoIs) and succinate dehydrogenase inhibitors (SDHIs) and the molecular mechanisms of resistance. Evolutionary process of fungicide resistance and management programme aiming to mitigate the emergence of resistance are also discussed in the review. The target protein of MBCs isβ-tubulin, and the resistance in phytopathogenic fungi is linked to point mutation in the target protein. Amino acid substitutions in target protein occur mainly at the positions 50, 167, 198, 200, and 240, and the most frequent mutation is amino acid 198. In general, only one substitution occurs in each resistant isolate. Resistant level varies among isolates with different substitutions. The target protein of DCFs has been unknown, the resistance may be correlated with point mutation in histidine kindnase (OS-related) genes. DMIs inhibit sterol 14α-demethylation step in biosynthesis of ergosterol and resistant mechanisms usually include point mutation ofCyp51 or over-expressions ofCyp51 and transporter genes. But point mutation inCyp51 is the major mechanism of DMI resistance. Different site mutations or even same site and same amino acid substitutions could lead to different resistance to triazoles. The number of point mutations in Cyp51varies among fungi, ranging from one mutation to several mutations and different mutations have an additive effect on DMI-resistance. QoIs affect the

  11. Ação do óleo essencial de Syzygium aromaticum (L. Merr. & L.M.Perry sobre as hifas de alguns fungos fitopatogênicos Action of Syzygium aromaticum (L. Merr. & L.M.Perry essential oil on the hyphae of some phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    A.R.T Costa

    2011-01-01

    Full Text Available Atualmente o uso de métodos alternativos para o controle de doenças e pragas na agricultura, visando minimizar os danos ao meio ambiente e à saúde pública é uma prática reconhecida e necessária. Este trabalho objetivou investigar a ação do óleo essencial de Syzygium aromaticum (L. Merr. & L.M.Perry sobre o crescimento micelial in vitro dos fungos fitopatogênicos Rhizoctonia solani, Fusarium solani, Fusarium oxysporum e Macrophomina phaseolina. A análise por cromatografia gasosa acoplada com espectrometria de massa possibilitou a identificação de eugenol (83,6%, acetato de eugenila (11,6% e cariofileno (4,2%. A avaliação microscópica dos micélios dos fungos evidenciou diversas alterações morfológicas, como a presença de vacúolos, desorganização dos conteúdos celulares, diminuição na nitidez da parede celular, intensa fragmentação e menor turgência das hifas. O óleo essencial de cravo apresentou atividade fungicida na concentração de 0,15% sobre o crescimento de R. solani, F. oxysporum e F. solani, entretanto não demonstrou essa atividade sobre M. phaseolina. Esses resultados indicam perspectivas favoráveis para posterior uso do óleo de cravo no controle desses fitopatógenos na agricultura.Currently, the use of alternative methods to control diseases and pests in agriculture has been a recognized and necessary practice to minimize damages to the environment and public health. This study aimed to investigate the action of clove [Syzygium aromaticum (L. Merr. & L.M.Perry] essential oil on the in vitro mycelial growth of the phytopathogenic fungi Rhizoctonia solani, Fusarium solani, Fusarium oxysporum and Macrophomina phaseolina. Analysis by gas chromatography-mass spectrometry allowed the identification of eugenol (83.6%, eugenyl acetate (11.6% and caryophyllene (4.2%. Microscopic evaluation of mycelia showed several morphological changes such as presence of vacuoles, cell content disorganization, decreased

  12. In vitro effect of Bacillus thuringiensis strains and Cry proteins in phytopathogenic fungi of paddy rice-field Efeito in vitro de cepas e proteínas Cry de Bacillus thuringiensis em fungos fitopatogênicos da cultura do arroz irrigado

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    Neiva Knaak

    2007-09-01

    Full Text Available Cry1Ab and Cry1Ac strains and proteins synthesized by Bacillus thuringiensis thuringiensis and B. thuringiensis kurstaki were assessed in the following phytopathogens: Rhizoctonia solani,Pyricularia grisea,Fusarium oxysporum and F. solani, which had their micelial growth decreased after incubation in the presence of the bacterial strains. As to Cry proteins, there were no inhibition halo development in the assessed concentrations.As cepas e proteínas Cry1Ab e Cry1Ac sintetizadas por Bacillus thuringiensis thuringiensis e B. thuringiensis kurstaki, foram avaliadas nos fitopatógenos: Rhizoctonia solani,Pyricularia grisea,Fusarium oxysporum e F. solani, os quais tiveram seu crescimento micelial reduzido após a incubação na presença das cepas bacterianas. Em relação às proteínas Cry, não houve formação de halo de inibição nas concentrações avaliadas.

  13. Fermentation Conditions Optimization of Antagonistic Bacteria B.velezensis 6-61Against Verticillium dahliae Kleb%棉花黄萎病拮抗细菌B.velezensis6-61拮抗蛋白发酵条件的优化

    Institute of Scientific and Technical Information of China (English)

    李红亚; 李术娜; 朱宝成

    2011-01-01

    为了提高棉花黄萎病拮抗细菌B.velezensis 6-61活性物质的产量,为进一步开展6-61菌株的后续研究奠定基础,对6-61菌株产生的活性物质的性质进行了分析,并对其发酵条件进行了优化研究.采用有机溶剂萃取、硫酸铵沉淀对菌株6-61发酵液进行处理并检测抑茵活性,初步确定了该拮抗物质为蛋白质.采用单因素试验对6-61菌株的发酵培养基所需的碳源、氮源、无机盐进行了筛选,并采用L16(44)正交实验对棉花黄萎病拮抗细菌6-61的发酵培养基组成以及发酵培养条件进行了优化.最终优化后的培养基组成为:麦芽糖3%、大豆蛋白胨3%、NaCl 0.01%、KCl 0.01%;最佳发酵条件:pH 7.0、接种量8%、种龄16h、装瓶量30 mL/250 mL.在优化后的培养基及培养条件下进行发酵,拮抗细菌6-61的发酵液抑茵圈直径可达到36 mm.与其他一些常见培养基的发酵效果相比较,在优化后的发酵条件下6-61茵株拮抗物质的产量明显高于其他常见培养基.%In order to improve the yield of antagonistic substance produced by B. Velezensis 6-61 which has antagonistic activity against Verticillium dahliae Kleb, the analysis of the properties of antagonistic substance and fermentation conditions optimization of stain 6-61 were studied to provide a basis for further study of the antagonistic mechanism. First, the fermented broth of strain 6-61 was treated by two following methods: extraction with organic solvent and precipitation by ammonium sulfate respectively. The results indicated that the component of strain 6-61 responsible for antagonistic activity was considered as protein. Then, the optimum fermentation medium ingredients, such as carbon sources, nitrogen sources and inorganic salts, were determined by using the single-factor methods. And the optimal medium compositions and other fermentation conditions of antagonistic bacteria 6-61 were optimized through Li6(44) orthogonal

  14. The pathogenic diversity and the genetic diversity of cotton Verticillium dahliae in the Yangtze River Basin%长江流域棉花黄萎病菌的致病力多样性和遗传多样性分析

    Institute of Scientific and Technical Information of China (English)

    金利容; 万鹏; 黄薇; 杨晓华

    2016-01-01

    Cotton Verticillium wilt is an important disease in the world, and a serious threat to cotton production.The four provinces, Hubei Province, Hunan Province, Jiangxi Province and Anhui Province, lie in the Yangtze River Basin.More than three hundreds isolates were collected and 67 isolates were selected from different cities and counties as the tested strains, then the cultural character, the pathogenic diversity and genetic diversity of the 67 strains were analyzed.The results showed that the high virulent types, the medium virulent types and the weak virulent types accounted for 35.82%, 43.28% and 17.91% respectively.The microsclerotia types, the intermediate types and the hyphae types acounted for 38.81%, 22.39% and 38.81%respectively.Further analysis showed that there was no correlation between the pathogenicity, the cultural character and the strain origin.However, there were some correlations between the pathogenicity and the cultural character.Most of high virulent strains belonged to microsclerotia types and the weak virulent strains dominate hyphae types.Fifteen pairs of RAPD primers were gained for DNA amplification, cluster analysis showed that the genetic diversity had no significant correlation with the strain origin.Furthermore, the genetic diversity of the strains had no significant correlation with the pathogenicity and the cultural character.%棉花黄萎病是棉花上的重要病害,严重威胁着棉花的产量.从长江流域4个省即湖北省、湖南省、江西省和安徽省采集300多个菌株,选择各省不同县市的67个菌株,对这67个供试菌株进行培养性状的鉴定、致病力的测定以及遗传多样性的分析,结果表明,在67个供试菌株中,就致病力类型而言,强致病力类型、中等致病力类型和弱致病力类型分别占35.82%、43.28%和17.91%.就培养性状类型而言,菌核型、中间型和菌丝型分别占38.81%、22.39%和38.81%.菌株的

  15. In vitro inhibition of pathogenic Verticillium dahliae, causal agent of ...

    African Journals Online (AJOL)

    SAM

    2014-08-13

    Aug 13, 2014 ... Combined analysis of wilt incidence resulting from using two inoculation methods for ... isolates consisting of 21 isolates isolated from potato soils from seven districts of Guizhou, 11 isolates .... were used as test plants.

  16. 沼液对植物病害的防治效果及机理研究I:对植物病原真菌的抑制效果及抑菌机理初探%Biocontrol Effect and Mechanism of Biogas Slurry on Plant Disease I :Primary Study of Growth Inhibition Effects and Mechanism on Phytopathogen Fungi

    Institute of Scientific and Technical Information of China (English)

    马艳; 李海; 常志州; 徐跃定; 张建英

    2011-01-01

    effect on spore germination of F. axysporurnfsp. fragariae with different degree. The inhibition effect of BS coming from pig manure and its sterilized filter was better than that of cow manure. Inhibition effect of both BS on five phytopathogens of strawberry showed significant difference:growth inhibition rate with 73%~87% on F. axysporurn fsp. fragariae, Colletotrichurn gloeosporioides (Penz.) Sacc. and Verticilliurn dahliae Kelb was obtained, while to Phtophthora capsici,we got growth inhibition rate with 38% and no inhibition effect on Pythiurn aphaniderrnaturn. Anti-microorganisms survived in BS played an important role in mycelia growth inhibition of phytopathogens and disease control of plant in consequence.

  17. Selection of endophytic fungi from comfrey (Symphytum officinale L. for in vitro biological control of the phytopathogen Sclerotinia sclerotiorum (Lib. Seleção de fungos endofíticos de confrei (Symphytum officinale L., buscando controle biológico in vitro do fitopatógeno Sclerotinia sclerotiorum (Lib.

    Directory of Open Access Journals (Sweden)

    Rafaeli Rocha

    2009-03-01

    Full Text Available Biological control consists of using one organism to attack another that may cause economic damage to crops. Integrated Pest Management (IPM is a very common strategy. The white mold produced by Sclerotiniasclerotiorum (Lib. causes considerable damage to bean crops. This fungus is a soil inhabitant, the symptoms of which are characterized by water-soaked lesions covered by a white cottony fungal growth on the soil surface and/or the host plant. Possible biological control agents taken from plants are being investigated as phytopathogen inhibitors. These are endophytic microorganisms that inhabit the intercellular spaces of vegetal tissues and are often responsible for antimicrobial production. The objective of the present study was to select endophytic fungi isolated from comfrey (Symphytumofficinale L. leaves with in vitro antagonist potential against the phytopathogenic fungus S. sclerotiorum. Twelve isolates of endophytic fungi and a pathogenic strain of S. sclerotiorum were used in the challenge method. With the aid of this method, four endophytes with the best antagonistic activity against S. sclerotiorum were selected. Pathogen growth inhibition zones were considered indicative of antibiosis. The percentages of pathogenic mycelia growth were measured both with and without the antagonist, resulting in growth reductions of 46.7% to 50.0% for S. sclerotiorum. These analyses were performed by evaluating the endophytic/pathogenic mycelia growth in mm/day over an eight-day period of antagonistic tests.O controle biológico consiste no uso de organismos que atacam outros que causam danos a culturas de plantas. Esta é uma estratégia muito utilizada no Controle Integrado de Pragas (CIP. O mofo branco, causado por Sclerotiniasclerotiorum (Lib., causa danos em culturas de feijão. Este fungo é encontrado no solo e seus sintomas são caracterizados por lesões úmidas cobertas por micélios algodonosos, crescidos a partir do solo e/ou da planta

  18. Potencial fungitóxico do óleo essencial de Piper hispidinervum (pimenta longa sobre os fungos fitopatogênicos Bipolaris sorokiniana, Fusarium oxysporum e Colletotrichum gloeosporioides Fungitoxic potential of the essential oil the Piper hispidinervum (long-pepper against phytopathogenic fungi Bipolaris sorokiniana, Fusarium oxysporum e Colletotrichum gloeosporioides

    Directory of Open Access Journals (Sweden)

    Lidiany Mendonça Zacaroni

    2009-03-01

    Full Text Available O objetivo deste trabalho foi avaliar a atividade fungicida in vitro do óleo essencial das folhas de Piper hispidinervum sobre Bipolaris sorokiniana, Fusarium oxysporum e Colletotrichum gloeosporioides. Para os ensaios biológicos, empregou-se o teste bioanalítico in vitro utilizando as concentrações de 100, 200, 500, 1000, 1500 e 2000 µg.mL-1 do óleo essencial. Estas foram incorporadas no meio de cultura BDA (batata-dextrose-ágar para avaliação do crescimento ou inibição micelial. O delineamento estatístico utilizado foi o inteiramente casualizado, com quatro repetições. Na concentração de 200 µg.mL-1, observou-se uma inibição total do fitopatógeno Bipolaris sorokiniana enquanto que, para o Fusarium oxysporum e o Colletotrichum gloeosporioides esta ocorreu na concentração de 1000 µg.mL-1.The objective of this study was to evaluate the in vitro antifungal activity of the essential oil of the leaves of Piper hispidinervum against Bipolaris sorokiniana, Fusarium oxysporum and Colletotrichum gloeosporioides. For the biological tests, using the bioanalitic test in vitro the concentrations of 100, 200, 500, 1000, 1500 and 2000 µg.mL-1 the essential oil. This were incorporated into PDA (potato dextrose agar medium in order to evaluate fungal mycelial growth or inhibition. The statistic design used was completely randomized, with four replicates. In the concentration of 200 µg.mL-1, observed inhibited complete the phytopathogens Bipolaris sorokiniana while the Fusarium oxysporum and Colletotrichum gloeosporioides this is occurred in the concentration of 1000 µg.mL-1.

  19. Atividade antimicrobiana de óleos essenciais no controle de alguns fitopatógenos fúngicos in vitro e no tratamento de sementes Antimicrobial activity of essential oils on the in vitro control of some fungal phytopathogens and on seed treatment

    Directory of Open Access Journals (Sweden)

    T. Hillen

    2012-01-01

    Full Text Available Este trabalho verificou o efeito dos óleos essenciais (OE extraídos de Eremanthus erythropappus (candeia, Cymbopogon martinii (palmarosa e de Rosmarinus officinalis (alecrim no crescimento micelial de alguns fitopatógenos fúngicos e no tratamento de sementes de milho, soja e feijão. No teste in vitro, alíquotas de 20, 40, 60, 100, 200, 500 e 1000 μL de cada um dos óleos essenciais foram distribuídas na superfície do meio de cultura. Posteriormente, discos de meio de cultura com micélio de Alternaria carthami, Alternaria sp. e Rhizoctonia solani foram transferidos para o centro de cada placa. O crescimento foi mensurado e calculada a taxa de inibição do crescimento micelial (ICM. Para verificar o efeito dos OE na germinação das sementes utilizou-se a aplicação deles por fumigação. Foi avaliada a percentagem de sementes germinadas e a incidência de patógenos nas sementes. Sobre o crescimento micelial, o óleo de palmarosa inibiu completamente todos os patógenos fúngicos, independentemente da concentração. Já os óleos de candeia e alecrim foram melhores quando foram adicionadas alíquotas superiores a 200 μL. Os óleos influenciaram diferentemente a germinação e a sanidade das sementes de milho, soja e feijão.This study aimed to verify the effect of essential oils (EO extracted from Eremanthus erythropappus ("candeia" Cymbopogon martinii ("palmarosa" and Rosmarinus officinalis (rosemary on the mycelial growth of some fungal phytopathogens, as well as on the treatment of corn, soybean and bean seeds. In the in vitro test, aliquots of 20, 40, 60, 100, 200, 500 and 1000 μL of each essential oil were distributed on the surface of the culture medium. Then, discs of culture medium with mycelium of Alternaria carthami, Alternaria sp and Rhizoctonia solani were transferred to the center of each plate. Growth was measured and the mycelial growth inhibition rate (MGI was calculated. To verify the effect of EO on seed

  20. Myconanoparticles: synthesis and their role in phytopathogens management

    Science.gov (United States)

    Alghuthaymi, Mousa A.; Almoammar, Hassan; Rai, Mahindra; Said-Galiev, Ernest; Abd-Elsalam, Kamel A.

    2015-01-01

    Nanotechnology can offer green and eco-friendly alternatives for plant disease management. Apart from being eco-friendly, fungi are used as bio-manufacturing units, which will provide an added benefit in being easy to use, as compared to other microbes. The non-pathogenic nature of some fungal species in combination with the simplicity of production and handling will improve the mass production of silver nanoparticles. Recently, a diverse range of fungi have been screened for their ability to create silver nanoparticles. Mycosynthesis of gold, silver, gold–silver alloy, selenium, tellurium, platinum, palladium, silica, titania, zirconia, quantum dots, usnic acid, magnetite, cadmium telluride and uraninite nanoparticles has also been reported by various researchers. Nanotechnological application in plant pathology is still in the early stages. For example, nanofungicides, nanopesticides and nanoherbicides are being used extensively in agriculture practices. Remote activation and monitoring of intelligent nano-delivery systems can assist agricultural growers of the future to minimize fungicides and pesticides use. Nanoparticle-mediated gene transfer would be useful for improvement of crops resistant to pathogens and pest. This review critically assesses the role of fungi in the synthesis of nanoparticles, the mechanism involved in the synthesis, the effect of different factors on the reduction of metal ions in developing low-cost techniques for the synthesis and recovery of nanoparticles. Moreover, the application of nanoparticles in plant disease control, antimicrobial mechanisms, and nanotoxicity on plant ecosystem and soil microbial communities has also been discussed in detail. PMID:26019636

  1. Antifungal activity of rice straw extract on some phytopathogenic fungi

    African Journals Online (AJOL)

    user

    2012-09-04

    Sep 4, 2012 ... The antifungal activity of allelochemicals extracted from rice straw on the radial growth rate and the activity of .... laurel, avocado and ginger extracts have strong ... composition and antifungal activity of essential oils of seven.

  2. Comparative Analyses of Exoproteinases Produced by Three Phytopathogenic Microorganisms

    Directory of Open Access Journals (Sweden)

    Tatiana A. Valueva

    2011-01-01

    Full Text Available Proteinases secreted by the oomycete Phytophthora infestans (Mont. de Bary, Rhizoctonia solani, and Fusarium culmorum belonging to different families of fungi have been studied to determine if the exoenzyme secretion depends on the environmental conditions and the phylogenetic position of the pathogen. The substrate specificity of the extracellular proteinases of F. culmorum, R. solani, and P. infestans and their sensitivity to the action of synthetic and protein inhibitors suggest that they contain trypsin-like and subtilisin-like enzymes regardless of culture medium composition. The relation of trypsin-like and subtilisin-like enzymes is dependent on the culture medium composition, especially on the form of nitrogen nutrition, particularly in the case of the exoenzymes secreted by R. solani. Phylogenetic analyses have shown that the exoproteinase set of ascomycetes and oomycetes has more similarities than basidiomycetes although they are more distant relatives. Our data suggests that the multiple proteinases secreted by pathogenic fungi could play different roles in pathogenesis, increasing the adaptability and host range, or could have different functions in survival in various ecological habitats outside the host.

  3. Glycyrrhiza glabra extract protects plants against important phytopathogenic fungi.

    Science.gov (United States)

    Schuster, C; Konstantinidou-Doltsinis, S; Schmitt, A

    2010-01-01

    In previous investigations an ethanolic plant extract from Glycyrrhiza glabra (2.5% w/v) showed 100% efficacy against late blight (Phytophthora infestans) on detached tomato leaves. Based on these findings, the objective of this work was to investigate the effect of this extract against different important plant pathogenic fungi. Tests were carried out on potted plants. Against P. infestans, efficacies of 75% and 58% were achieved on tomato and potato plants with 5% extract concentration, respectively. Against another Oomycete, Pseudoperonospora cubensis, on cucumber, application of a 2.5% extract led to an efficacy of above 90%. The EC50-value was calculated to be 0.5% In a trial on beans against bean rust (Uromyces appendiculatus), G. glabra extract (5% concentration) showed 92% efficacy. In contrast, against powdery mildew on cucumber (Podosphaera xanthii), no disease reduction was found. Overall, the results indicate a high potential for the extract of G. glabra to control a number of important plant pathogens.

  4. In Vitro antifungal potency of plant extracts against five phytopathogens

    Directory of Open Access Journals (Sweden)

    Ashwani Tapwal

    2011-12-01

    Full Text Available The antifungal activity of aqueous extract of Cannabis sativa, Parthenium hysterophorus, Urtica dioeca, Polystichum squarrosum and Adiantum venustum was investigated against Alternaria solani, Alternaria zinniae, Curvularia lunata, Rhizoctonia solani and Fusarium oxysporum at different concentrations (5, 10, 15 and 20%. At 20%, maximum antifungal potential was observed with the extracts of C. sativa, which recorded excellent inhibitory activity against C. lunata (100%, A. zinniae (59.68%, followed by leaf extract of P. hysterophorus (50% against A. solani. The application of botanical extracts for disease management could be less expensive, easily available, non-polluting and eco-friendly.

  5. Activity of Azoxystrobin and SHAM to Four Phytopathogens

    Institute of Scientific and Technical Information of China (English)

    JIN Li-hua; CHEN Yu; CHEN Chang-jun; WANG Jian-xin; ZHOU Ming-guo

    2009-01-01

    The study was conducted to make clear the activity of azoxystrobin to 4 plant pathogens and the synergistic effects of salicylhydroxamic acid (SHAM), which acted on the alternative oxidase. It was also conducted to be aware of the mechanism of azoxystrobin in inhibition on mycelial respiration and the influence of SHAM. The activity test of azoxystrobin and SHAM was carried out with a mycelial linear growth test and spore germination test. Other related biological properties were also observed. Inhibition of azoxystrobin and SHAM on 4 pathogens was determined by using SP-Ⅱ oxygraph system. Azoxystrobin inhibited mycelial growth in Colletotrichum capsici, Botrytis cinerea, Rhizoctonia solani, and Magnaporthe grisea, respectively; it also inhibited conidia germination, and conidia production in C. capsici, B. cinerea M. grisea, and sclerotia formation in R. solani. Moreover, it created stayed pigment biosynthesis in C. capsici and M. grisea somehow. Salicylhydroxamic acid enhanced inhibition by azoxystrobin. An oxygen consuming test of the mycelia showed that azoxystrobin inhibited all the 4 fungi's respiration in the early stages. With the concentration rising up, the effectiveness increased. However, as time went on, the respiration of the mycelia treated with fungicides recovered and SHAM could not inhibit the oxygen consuming. This reaction between the mycelia and the fungicides appeared not to initiate alternative respiration but rather the other mechanism created a lack of efficacy.

  6. Inhibition of growth of some phytopathogenic and mycotoxigenic ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-08-18

    Aug 18, 2008 ... Whatman no 1 filter paper and the filtrate obtained was freeze-dried to complete ... analysis on a Perkin-Elmer 2000 FT-IR instrument set at 7800-370 cm-1 in a single ... revealed the presence of aluminium oxide (Al2O3), cal-.

  7. Microscopic phytopathogenic fungi rare and new for Poland

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    Małgorzata Ruszkiewicz

    2014-08-01

    Full Text Available The paper presents a list of 36 rare species of fungi with remarks about their morphology and distribution in Poland. Three of the species are new for Poland: Romularia asplenii Jaap, R. concomitans Ell. et Holw. and Ascochyta actoeae (Bres. J. J. Davis. Altogether, 13 parasitic species occur on the plans which have not been known as their hosts in Poland, so far, 19 are rare on the listed hosts. The fungi were collected in the area of projected Jurassic National Park, Częstochowa Upland.

  8. Phytotoxic terpenes produced by phytopathogenic fungi and allelopathic plants.

    Science.gov (United States)

    Cimmino, Alessio; Andolfi, Anna; Evidente, Antonio

    2014-03-01

    This review is about the isolation as well as chemical and biological characterization of simple and complex mono-, sesqui-, di-, sester- and tri-terpenes produced by fungal pathogens of agrarian and forest plants and by some allelopathic plants. In several cases, the structure activity relationships are also discussed, as well as their potential application in agriculture as natural safe herbicides, fungicides and bactericides. Furthermore, the potential application of some fungal terpenes as anticancer compounds with a new mode of action is also discussed.

  9. Multilocus sequence analysis of phytopathogenic species of the genus Streptomyces

    Science.gov (United States)

    The identification and classification of species within the genus Streptomyces is difficult because there are presently 576 validly described species and this number increases every year. The value of the application of multilocus sequence analysis scheme to the systematics of Streptomyces species h...

  10. Fitodefensivos em plantas medicinais: macromoléculas hidrofílicas de folhas de mil folhas (Achillea millefolium L. inibem o crescimento in vitro de bactérias fitopatogênicas Agrochemicals in medicinal plants: hydrophilic macromolecules from leaves of "mil folhas" (Achillea millefolium L. inhibit in vitro growth of phytopathogenic bacteria

    Directory of Open Access Journals (Sweden)

    N.G. Tessarollo

    2013-01-01

    Full Text Available Extratos aquosos da planta medicinal Achillea millefolium contêm macromoléculas de interesse para desenvolver fitodefensivos para a agricultura. Duas frações de mil folhas foram obtidas por ultrafiltração, E1 (contendo moléculas maiores que 30 kDa, e E3 (peptídeos entre 1 e 10 kDa que inibiram o crescimento das bactérias fitopatogênicas Ralstonia solanacearum, gram-negativa, e Clavibacter michiganensis subsp. michiganensis, gram-positiva, com dependência de concentração. Os valores de concentração inibitória mínima (CIM para ambos os extratos e bactérias foram baixos, entre 20 e 80µM. A CIM relativa à proteína total evidenciou a presença de macromoléculas muito ativas em E3, embora com baixa concentração proteica. E3 se aplica à prospecção de peptídeos antimicrobianos. Estimar a CIM relativa à quantidade de amostra vegetal valorizou o potencial antimicrobiano natural de E1, que contém alta concentração proteica. E1e E3 se aplicam ao desenvolvimento de fitodefensivos para uso biotecnológico. A ultrafiltração fracionou as amostras de forma nativa, rápida, e com baixo custo; além de dessalinizar, clarificar, purificar, e concentrar E1 e E3. Esse estudo inédito sobre a separômica e a ação antimicrobiana de extratos macromoleculares aquosos de mil folhas sugere que plantas cicatrizantes podem apresentar grande potencial para desenvolver fitodefensivos agrícolas naturais não danosos, à semelhança de medicamentos fitoterápicos.Aqueous extracts from the medicinal plant Achillea millefolium contain macromolecules of interest to develop agrochemicals for agriculture. Two fractions of "mil folhas" were obtained by ultrafiltration, E1 (containing molecules larger than 30 kDa and E3 (peptides between 1 and 10 kDa, which inhibited the growth of phytopathogenic bacteria Ralstonia solanacearum, gram-negative, and Clavibacter michiganensis subsp. michiganensis, gram-positive, concentration-dependent. The values of

  11. Benzimidazole-1,2,3-triazole hybrid molecules: synthesis and evaluation for antibacterial/antifungal activity.

    Science.gov (United States)

    Ouahrouch, Abdelaaziz; Ighachane, Hana; Taourirte, Moha; Engels, Joachim W; Sedra, My Hassan; Lazrek, Hassan B

    2014-10-01

    A novel series of hybrid molecules 4a-i and 5a-i were prepared by condensation of 4-(trimethylsilylethynyl)benzaldehyde 1 with substituted o-phenylenediamines. These in turn were reacted with 2-(azidomethoxy)ethyl acetate in a Cu alkyne-azide cycloaddition (CuAAC) to generate the 1,2,3-triazole pharmacophore under microwave assistance. The newly synthesized compounds were examined for their in vitro antimicrobial activities against Gram-positive and Gram-negative bacteria and the phytopathogenic fungi Verticillium dahliae and Fusarium oxysporum f. sp. albedinis. 2-((4-(4-(5-Trifluoromethyl benzimidazol-2-yl)phenyl)-1,2,3-triazol-1-yl)methoxy)ethanol 5e showed a moderate inhibition of 30% in the Foa sporulation test.

  12. Molecular Detection of Verticillium albo-atrum by PCR Based on Its Sequences

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticilliumalbo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer (ITS) sequences ofVerticilliun spp., a pair of species-specific primers, Vaa1/Vaa2, was synthesized. After screening 17 isolates of V. albo-atrum, 121 isolates from the Ascomycota, Basidiomycota, Deuteromycota, and Oomycota, the Vaa1/Vaa2 primers amplifiedonly a single PCR band of approximately 330 bp from V. albo-atrum. The detection sensitivity with primers Vaa1/Vaa2 was10 fg of genomic DNA. Using ITS1/ITS4 as the first-round primers, combined with Vaa1/Vaa2, the nested PCR procedureswere developed, and the detection sensitivity increased 1 000-fold to 10 ag. The detection sensitivity for the soil pathogenswas 100-conidiag-1 soil. The PCR-based methods developed here could simplify both plant disease diagnosis and pathogen monitoring as well as guide plant disease management.

  13. Pathogenicity and virulence of the two Dutch VCGs of Verticillium dahliae to woody ornamentals

    NARCIS (Netherlands)

    Goud, J.C.; Termorshuizen, A.J.

    2002-01-01

    Two experiments were performed in two consecutive years to test whether isolates of different vegetative compatibility groups (VCGs) differ in their ability to cause disease in woody ornamentals, to study the host specificity of the isolates and to get an insight into disease development in woody

  14. Verticillium-verwelkingsziekte in bomen: detectie, voorspelling van aantasting en beheersing van de ziekte

    NARCIS (Netherlands)

    Goud, J.C.

    2004-01-01

    Resultaten van promotie-onderzoek, waarbij in twee veldproeven de relatie werd vastgesteld tussen verwelkingsziekte in Noorse esdoorn (Acer platanoides) en trompetboom (Catalpa bignonioides) en besmettingsgraad van de bodem. Tevens werd de effectiviteit van biologische grondontsmetting onderzocht al

  15. Genomic Analysis of Verticillium Wilt Resistance and Drought Tolerance in Alfalfa

    Science.gov (United States)

    Alfalfa (Medicago sativa L.) is the fourth largest crop in the United States. Changing trends to multipurpose uses increases demand for alfalfa. However, the production of alfalfa is challenged by endemic and emerging diseases and adverse environmental factors. Identification of genes/loci controlli...

  16. Diversity and biotransformative potential of endophytic fungi associated with the medicinal plant Kadsura angustifolia.

    Science.gov (United States)

    Huang, Qian; An, Hongmei; Song, Hongchuan; Mao, Hongqiang; Shen, Weiyun; Dong, Jinyan

    2015-01-01

    This study investigated the diversity and host component-transforming activity of endophytic fungi in medicinal plant Kadsura angustifolia. A total of 426 isolates obtained were grouped into 42 taxa belonging to Fungi Imperfecti (65.96%), Ascomycota (27.00%), Zygomycota (1.64%), Basidiomycota (0.47%) and Mycelia Sterilia (4.93%). The abundance, richness, and species composition of endophytic assemblages were significantly dependent on the tissue and the sampling site. Many phytopathogenic species associated with healthy K. angustifolia were found prevalent. Among them, Verticillium dahliae was dominant with 16.43% abundance. From 134 morphospecies selected, 39 showed remarkable biocatalytic activity and were further identified as species belonging to the genera Colletotrichum, Eupenicillium, Fusarium, Hypoxylon, Penicillium, Phomopsis, Trametes, Trichoderma, Umbelopsis, Verticillium and Xylaria on the basis of the sequence analysis of the internal transcribed spacer (ITS1-5.8S-ITS2). The results obtained in this work show that K. angustifolia is an interesting reservoir of pathogenic fungal species, and could be a community model for further ecological and evolutionary studies. Additionally, the converting potency screening of some endophytic fungi from this specific medicinal plant may provide an interesting niche on the search for novel biocatalysts. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  17. Synthetic blends of volatile, phytopathogen-induced odorants can be used to manipulate vector behavior.

    Directory of Open Access Journals (Sweden)

    Alexander eAksenov

    2014-12-01

    Full Text Available Volatile organic compounds (VOCs are emitted from all plants and these VOCs are important means of communication between plants and insects. It has been documented that pathogen infections alter VOC profiles rendering infected plants more attractive to specific vectors transmitting these pathogens than uninfected plants, thus potentially aiding in pathogen propagation. Mimicking these chemical cues might enable insect attraction away from the plant or disruption of host finding behavior of the vector. However, the practical implications have not been fully explored. We used citrus, Diaphorina citri and huanglongbing (HLB as a model host-vector-disease system because HLB threatens citrus production worldwide and is similar to other critical diseases of food crops, such as Zebra Chip affecting potato. We formulated a synthetic chemical blend using selected HLB-specific biomarker compounds, and tested the blend with the Attenu assay system for chemosensory proteins. The Attenu assay system is a procedure that identifies interactions between insect chemosensory proteins and their ligands. We found that an equimolar mixture of compounds mimicking the volatile profile of HLB-infected citrus bound chemosensory proteins. Further investigation of this blend in laboratory behavioral assays resulted in development of a synthetic lure that was more attractive to D. citri than natural citrus tree volatiles. This strategy could provide a new route to produce chemical lures for vector population control for a variety of plant and/or animal systems and it may result in the development of a practical lure for monitoring vectors of disease, such as D. citri.

  18. Unraveling the protein network of tomato fruit in response to necrotrophic phytopathogenic Rhizopus nigricans.

    Directory of Open Access Journals (Sweden)

    Xiaoqi Pan

    Full Text Available Plants are endowed with a sophisticated defense mechanism that gives signals to plant cells about the immediate danger from surroundings and protects them from pathogen invasion. In the search for the particular proteins involved in fruit defense responses, we report here a comparative analysis of tomato fruit (Solanum lycopersicum cv. Ailsa Craig infected by Rhizopus nigricans Ehrenb, which is a significant contributor to postharvest rot disease in fresh tomato fruits. In total, four hundred forty-five tomato proteins were detected in common between the non-infected group and infected tomato fruit of mature green. Forty-nine differentially expressed spots in 2-D gels were identified, and were sorted into fifteen functional groups. Most of these proteins participate directly in the stress response process, while others were found to be involved in several equally important biological processes: protein metabolic process, carbohydrate metabolic process, ethylene biosynthesis, and cell death and so on. These responses occur in different cellular components, both intra- and extracellular spaces. The differentially expressed proteins were integrated into several pathways to show the regulation style existing in tomato fruit host. The composition of the collected proteins populations and the putative functions of the identified proteins argue for their roles in pathogen-plant interactions. Collectively results provide evidence that several regulatory pathways contribute to the resistance of tomato fruit to pathogen.

  19. Whole cell fatty acid analysis as a tool for classification of phytopathogenic pseudomonas bacteria.

    NARCIS (Netherlands)

    Janse, J.D.

    1992-01-01

    In this thesis some members of the plant pathogenic bacterial genus Pseudomonas have been studied. Conventional morphological, biochemical, physiological and pathogenitcity tests as well as a 'finger-print' technique, viz. automated whole cell fatty acid analysis, were used. The taxonomy of the plan

  20. Detection and characterization of a novel Gammapartitivirus in the phytopathogenic fungus Colletotrichum acutatum strain HNZJ001.

    Science.gov (United States)

    Zhong, Jie; Chen, Dan; Lei, Xiang Hua; Zhu, Hong Jian; Zhu, Jun Zi; Da Gao, Bi

    2014-09-22

    Spherical virus-like particles about 40nm in diameter were observed under transmission electron microscope (TEM) and two dsRNA bands (dsRNA-1 and dsRNA-2) were detected on agarose gel after extraction from the mycelial preparation of a Colletotrichum acutatum strain HNZJ001 that isolated from an anthracnose lesion on immature pepper fruit. The complete nucleotide sequences of the dsRNAs were determined. DsRNA-1 (1762 nt) and dsRNA-2 (1381 nt) each contained a single open reading frame and potentially encoded 62 kDa and 40 kDa proteins, respectively. The 62 kDa protein showed similarity to the RNA-dependent RNA polymerase (RdRp) of partitiviruses, while the 40 kDa product had no significant similarity to any published capsid protein throughout all databases, besides of low homology with the hypothetical "capsid" protein of a few partitiviruses in fungus Ustilaginoidea virens. Genome comparison and phylogenetic analysis indicated that the virus is closely related to the mycovirus in the family Partitiviridae. The results suggested a novel two-segment dsRNA virus be detected. We name it Colletotrichum acutatum partitivirus 1 (CaPV1). RT-PCR detection, using a primer pair based on the RdRp of the dsRNA-1 showed very high efficiency of CaPV1 transmission into the progenies of the fungus. Virus curing and fungal phenotype observation for evaluation of the impact of CaPV1 in host fungus were also carried out.

  1. Essential oils from Algerian species of Mentha as new bio-control agents against phytopathogen strains.

    Science.gov (United States)

    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.

  2. Growth and enzymatic responses of phytopathogenic fungi to glucose in culture media and soil

    Directory of Open Access Journals (Sweden)

    Beatriz de Oliveira Costa

    2012-03-01

    Full Text Available The effect of inoculation of Aspergillus flavus, Fusarium verticillioides, and Penicillium sp. in Dystrophic Red Latosol (DRL and Eutroferric Red Latosol (ERL soils with or without glucose on the total carbohydrate content and the dehydrogenase and amylase activities was studied. The fungal growth and spore production in culture medium with and without glucose were also evaluated. A completely randomized design with factorial arrangement was used. The addition of glucose in the culture medium increased the growth rate of A. flavus and Penicillium sp. but not of F. verticillioides. The number of spores increased 1.2 for F. verticillioides and 8.2 times for A. flavus in the medium with glucose, but was reduced 3.5 times for Penicillium sp. The total carbohydrates contents reduced significantly according to first and second degree equations. The consumption of total carbohydrates by A. flavus and Penicillium sp. was higher than the control or soil inoculated with F. verticillioides. The addition of glucose to soils benefited the use of carbohydrates, probably due to the stimulation of fungal growth. Dehydrogenase activity increased between 1.5 to 1.8 times (p <0.05 in soils with glucose and inoculated with the fungi (except F. verticillioides, in relation to soil without glucose. Amylase activity increased 1.3 to 1.5 times due to the addition of glucose in the soil. Increased amylase activity was observed in the DRL soil with glucose and inoculated with A. flavus and Penicillium sp. when compared to control.

  3. Resistances to an insect herbivore and a phytopathogen in Barbarea vulgaris

    DEFF Research Database (Denmark)

    Christensen, Stina

    pubescence; one has glabrous leaves and is therefore called G-type while the other has pubescent leaves and is called the P-type. The G-type is resistant to most genotypes of the flea beetle Phyllotreta nemorum as well as some other Brassicales specialists, and this resistance is conferred by saponins. The P...

  4. Scorpion toxins modify phytopathogenic fungus physiology. A possible source of new fungicides.

    Science.gov (United States)

    Joya, Galax; D'Suze, Gina; Salazar, Víctor; Rosales, Arnaldo; Sevcik, Carlos; Visbal, Gonzalo; Ferreira, André T S; Perales, Jonas

    2011-06-08

    Seven toxins (F1-F7) were purified from Tityus discrepans scorpion venom on a C18 HPLC column. The compounds were fungitoxic on Macrophomina phaseolina. The molecular masses of F1-F7 were (Da) 1061.1, 7328.8, 7288.3, 7268.5, 7104.6, 6924.6, and 6823.3, respectively. It is not known if F1 is a small peptide or some other kind of organic molecule. Compounds F2-F7 were peptides. The most potent was F7, with a minimal inhibition concentration of 0.4 μg/μL and a concentration for 50% inhibition of 0.13 μg/μL. Fungal esterase activity was abolished by F2, F3, and F5 and inhibited by 89, 60, 58, and 54% by F4, F6, F7, and F1, respectively. F1, F2, F5, and F7 induced an increase on hyphae chitin wall and septum thickness. Peptides F3-F6 induced efflux of the fluorescent dye Na-CoroNa Red complex from hyphae. Only F5 and F6 were inhibited by the prokaryote sodium channel blockers amiloride and mibefradil. Gas chromatography-mass spectrometry analysis suggested that F1, F5, F6, and F7 altered sterol biosynthesis either by inhibiting ergosterol biosynthesis or by producing ergosterol analogues. The peptides affect M. phaseolina viability by three mechanisms: decreasing esterase activity, altering Na(+) membrane permeability, and altering wall sterol biosynthesis. It seems that interfering with sterol synthesis is an important mechanism behind the effect of the fungicideal toxins. However, the antifungal effects at short times are indicative of a direct esterase inhibition, which, with the increased membrane leakiness to Na(+), makes the fungus inviable.

  5. Genome Sequences of Three Phytopathogenic Species of the Magnaporthaceae Family of Fungi.

    Science.gov (United States)

    Okagaki, Laura H; Nunes, Cristiano C; Sailsbery, Joshua; Clay, Brent; Brown, Doug; John, Titus; Oh, Yeonyee; Young, Nelson; Fitzgerald, Michael; Haas, Brian J; Zeng, Qiandong; Young, Sarah; Adiconis, Xian; Fan, Lin; Levin, Joshua Z; Mitchell, Thomas K; Okubara, Patricia A; Farman, Mark L; Kohn, Linda M; Birren, Bruce; Ma, Li-Jun; Dean, Ralph A

    2015-09-28

    Magnaporthaceae is a family of ascomycetes that includes three fungi of great economic importance: Magnaporthe oryzae, Gaeumannomyces graminis var. tritici, and Magnaporthe poae. These three fungi cause widespread disease and loss in cereal and grass crops, including rice blast disease (M. oryzae), take-all disease in wheat and other grasses (G. graminis), and summer patch disease in turf grasses (M. poae). Here, we present the finished genome sequence for M. oryzae and draft sequences for M. poae and G. graminis var. tritici. We used multiple technologies to sequence and annotate the genomes of M. oryzae, M. poae, and G. graminis var. tritici. The M. oryzae genome is now finished to seven chromosomes whereas M. poae and G. graminis var. tritici are sequenced to 40.0× and 25.0× coverage respectively. Gene models were developed by the use of multiple computational techniques and further supported by RNAseq data. In addition, we performed preliminary analysis of genome architecture and repetitive element DNA.

  6. Biochemical study of the extracellular aspartyl protease Eap1 from the phytopathogen fungus Sporisorium reilianum.

    Science.gov (United States)

    Mandujano-González, Virginia; Arana-Cuenca, Ainhoa; Anducho-Reyes, Miguel Ángel; Téllez-Jurado, Alejandro; González-Becerra, Aldo E; Mercado-Flores, Yuridia

    2013-12-01

    In this work, the extracellular protease Eap1 from Sporisorium reilianum was characterized in solid and liquid cultures using different culture media. The results showed that Eap1 was produced in all media and under all culture conditions, with the most activity in solid culture at an acidic pH of 3-5. Following purification, the 41 kDa protease demonstrated aspartyl protease activity. The enzyme was stable at a wide range of temperatures and pH values, but 45°C and pH 3 were optimal. The K(m) and V(max( values obtained were 0.69 mg/mL and 0.66 μmol/min, respectively, with albumin as the substrate. Eap1 degraded hemoglobin as well as proteins obtained from corn germ, roots, stems and slides at pH 3 and also had milk-clotting activity. Sequencing analysis showed that this protein has 100% similarity to the peptide sequence theoretically obtained from the sr11394 gene, which encodes an aspartyl protease secreted by S. reilianum.

  7. Antimicrobial and anti-pathogenic activity of some thioureides derivatives against Erwinia amylovora phytopathogenic strains.

    Science.gov (United States)

    Măruţescu, Luminiţa; Niţulescu, Mihai-George; Bucur, Marcela; Diţu, Lia-Mara; Mihăescu, Grigore; Lazăr, Veronica; Sesan, Tatiana

    2011-01-01

    A series of N-(1-methyl-1 Hpyrazole-4-carbonyl)-thiourea derivatives were assessed for their in vitro antimicrobial and anti-pathogenic activity against twenty-two strains of Erwinia amylovora isolated from different regions in Romania. The compounds were solubilised in dimethylsulfoxide and screened for their in vitro antimicrobial activity. The qualitative screening of the susceptibility spectra of various strains to the compounds was performed by adapted diffusion techniques (distribution of the tested compound solution directly on the solid medium previously seeded with the bacterial inoculums). The quantitative assay of the minimal inhibitory concentration (MIC, microg/mL) was based on liquid medium two-fold microdilutions. The subinhibitory concentrations of the tested substances were investigated for their influence on biofilm development on inert substrata. The present study showed that six new thiourea compounds exhibited a low antibacterial activity (MIC values > 500 microg/ml), but the subinhibitory concentrations inhibited the biofilm development on inert substrata. Thus, these results could suggest the usefulness of the tested compounds as control agents for preventing the first stage (colonization) of the infection with the fire blight pathogen.

  8. Mycosphaerella podagrariae-a necrotrophic phytopathogen forming a special cellular interaction with its host Aegopodium podagraria

    NARCIS (Netherlands)

    Simon, U.K.; Groenewald, J.Z.; Stierhof, Y.D.; Crous, P.W.; Bauer, R.

    2010-01-01

    We present a new kind of cellular interaction found between Mycosphaerella podagrariae and Aegopodium podagraria, which is remarkably different to the interaction type of the obligate biotrophic fungus Cymadothea trifolii, another member of the Mycosphaerellaceae (Capnodiales, Dothideomycetes, Ascom

  9. Mycosphaerella podagrariae - a necrotrophic phytopathogen forming a special cellular interaction with its host Aegopodium podagraria

    NARCIS (Netherlands)

    Simon, U.K.; Groenewald, J.Z.; Stierhof, Y.D.; Crous, P.W.; Bauer, R.

    2010-01-01

    We present a new kind of cellular interaction found between Mycosphaerella podagrariae and Aegopodium podagraria, which is remarkably different to the interaction type of the obligate biotrophic fungus Cymadothea trifolii, another member of the Mycosphaerellaceae (Capnodiales, Dothideomycetes, Ascom

  10. Mycosphaerella podagrariae - a necrotrophic phytopathogen forming a special cellular interaction with its host Aegopodium podagraria

    OpenAIRE

    Simon, U.K.; Groenewald, J.Z.; Stierhof, Y D; Crous, P.W.; Bauer, R

    2010-01-01

    We present a new kind of cellular interaction found between Mycosphaerella podagrariae and Aegopodium podagraria, which is remarkably different to the interaction type of the obligate biotrophic fungus Cymadothea trifolii, another member of the Mycosphaerellaceae (Capnodiales, Dothideomycetes, Ascomycota) which we have described earlier. Observations are based on both conventional and cryofixed material and show that some features of this particular interaction are better discernable after ch...

  11. The Influence of Learning on Host Plant Preference in a Significant Phytopathogen Vector, Diaphorina citri.

    Science.gov (United States)

    Stockton, Dara G; Martini, Xavier; Patt, Joseph M; Stelinski, Lukasz L

    2016-01-01

    Although specialist herbivorous insects are guided by innate responses to host plant cues, host plant preference may be influenced by experience and is not dictated by instinct alone. The effect of learning on host plant preference was examined in the Asian citrus psyllid, Diaphorina citri; vector of the causal agent of citrus greening disease or huanglongbing. We investigated: a) whether development on specific host plant species influenced host plant preference in mature D. citri; and b) the extent of associative learning in D. citri in the form of simple and compound conditioning. Learning was measured by cue selection in a 2-choice behavioral assay and compared to naïve controls. Our results showed that learned responses in D. citri are complex and diverse. The developmental host plant species influenced adult host plant preference, with female psyllids preferring the species on which they were reared. However, such preferences were subject to change with the introduction of an alternative host plant within 24-48 hrs, indicating a large degree of experience-dependent response plasticity. Additionally, learning occurred for multiple sensory modalities where novel olfactory and visual environmental cues were associated with the host plant. However, males and females displayed differing discriminatory abilities. In compound conditioning tasks, males exhibited recognition of a compound stimulus alone while females were capable of learning the individual components. These findings suggest D. citri are dynamic animals that demonstrate host plant preference based on developmental and adult experience and can learn to recognize olfactory and visual host plant stimuli in ways that may be sex specific. These experience-based associations are likely used by adults to locate and select suitable host plants for feeding and reproduction and may suggest the need for more tailored lures and traps, which reflect region-specific cultivars or predominate Rutaceae in the area being monitored.

  12. Toxicity, analgesic and sedative potential of crude extract of soil-borne phytopathogenic fungi Aspergillus flavus

    Directory of Open Access Journals (Sweden)

    Bashir Ahmad

    2016-11-01

    Full Text Available Background: Aspergillus flavus is one of the most abundant mold present around the world. The present study was conducted to investigate the acute toxicity, analgesic and sedative effect of the crude extract obtained from soil borne fungi A. flavus. Methods: The fungi was isolated from soil samples and identified morphologically and microscopically. The growth condition i.e. media, temperature, pH, and incubation period were optimized. In these optimized growth condition, A. flavus was grown in batch culture in shaking incubator. Crude contents were extracted by using ethyl acetate solvent. Crude secondary metabolites were screened for acute toxicity, analgesic and sedative effect. Results: Upon completion of the experiment, blood was collected from the tail vein of albino mice, and different haematological tests were conducted. White blood cells counts displayed a slight increase (10.6× 109/L above their normal range (0.8–6.8 × 109/L, which may be due to the increment in the number of lymphocytes or granulocytes. However, the percentage of lymphocytes was much lower (17.7%, while the percentage of the granulocytes was higher (61.4% than its normal range (8.6–38.9%. A reduction in the mean number of writhing in the different test groups was caused by the application of the crude ethyl acetate extract through the i.p. route at different doses (50, 100, and 150 mg/kg body weight. The results of our investigation showed the EtOAc extract of A. flavus can cause a significant sedative effect in open field. Conclusion: It was concluded from the present study that the A. flavus has the potential to produce bioactive metabolites which have analgesic and sedative effect.

  13. Proteome of the phytopathogen Xanthomonas citri subsp. citri: a global expression profile

    Directory of Open Access Journals (Sweden)

    Ferro Jesus A

    2010-11-01

    Full Text Available Abstract Background Citrus canker is a disease caused by Xantomonas citri subsp.citri (Xac, and has emerged as one of the major threats to the worldwide citrus crop because it affects all commercial citrus varieties, decreases the production and quality of the fruits and can spread rapidly in citrus growing areas. In this work, the first proteome of Xac was analyzed using two methodologies, two-dimensional liquid chromatography (2D LC and tandem mass spectrometry (MS/MS. Results In order to gain insight into the metabolism of Xac, cells were grown on two different media (NB - Nutrient Broth and TSE - Tryptone Sucrose broth enriched with glutamic acid, and proteins were proteolyzed with trypsin and examined by 2D LC-MS/MS. Approximately 39% of all predicted proteins by annotation of Xac were identified with their component peptides unambiguously assigned to tandem mass spectra. The proteins, about 1,100, were distributed in all annotated functional categories. Conclusions This is the first proteomic reference map for the most aggressive strain of Xanthomonas pathogen of all orange varieties. The compilation of metabolic pathways involved with bacterial growth showed that Xac expresses a complete central and intermediary metabolism, replication, transcription and translation machineries and regulation factors, distinct membrane transporters (ABC, MFS and pumps and receptors (MCP, TonB dependent and metabolites acquisition, two-component systems (sensor and regulatory components and response regulators. These data corroborate the growth curve in vitro and are the first reports indicating that many of these genome annotated genes are translated into operative in Xac. This proteomic analysis also provided information regarding the influence of culture medium on growth and protein expression of Xac.

  14. Trichoderma volatiles effecting Arabidopsis: from inhibition to protection against phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    Metwaly eRamadan

    2015-09-01

    Full Text Available Trichoderma species are present in many ecosystems and some strains have the ability to reduce the severity of plant diseases by activating various defence pathways via specific biologically active signaling molecules. Hence we investigated the effects of low molecular weight volatile compounds of Trichoderma asperellum IsmT5 on Arabidopsis thaliana. During co-cultivation of T. asperellum IsmT5 without physical contact to A. thaliana we observed smaller but vital and robust plants. The exposed plants exhibit increased trichome numbers, accumulation of defence-related compounds such as H2O2, anthocyanin, camalexin, and increased expression of defence related genes. We conclude that A. thaliana perceives the Trichoderma volatiles as stress compounds and subsequently initiates multilayered adaptations including activation of signaling cascades to withstand this environmental influence.The prominent headspace volatile of T. asperellum IsmT5 was identified to be 6-pentyl-α-pyrone, which was solely applied to A. thaliana to verify the growth and defence reactions. Most noticeable is that A. thaliana preexposed to 6PP showed significantly reduced symptoms when challenged with Botrytis cinerea and Alternaria brassicicola, indicating that defence-activated plants subsequently became more resistant to pathogen attack. Together, these results support that products that are based on Trichoderma volatiles have the potential being a useful biocontrol agent in agriculture.

  15. IN VITRO ANTIFUNGAL ACTIVITY OF ESSENTIAL OILS ON GROWTH OF PHYTOPATHOGENIC FUNGI

    Directory of Open Access Journals (Sweden)

    Jasenka Ćosić

    2010-12-01

    Full Text Available Eleven essential oils (clove, rosemary, cinnamon leaf, sage, scots pine, neroli, peppermint, aniseed, caraway, lavander, common thyme were tested for in vitro antifungal activity on twelve plant pathogenic fungi (Fusarium graminearum, F. verticillioides, F. subglutinans, F. oxysporum, F. avenaceum, Diaporthe helianthi, Diaporthe phaseolorum var. caulivora, Phomopsis longicolla, P. viticola, Helminthosporium sativum, Colletotrichum coccodes, Thanatephorus cucumeris. The results indicated that all oils except scots pine and neroli had antifungal activity against some or all tested fungi. The best antifungal activity had common thyme, cinnamon leaf, clove and aniseed oils. When compared to control, scots pine, neroli and sage oils stimulated mycelium growth of some investigated fungi.

  16. Study of the mode of action of a polygalacturonase from the phytopathogen Burkholderia cepacia

    DEFF Research Database (Denmark)

    Massa, C.; Clausen, Mads Hartvig; Stojan, J.

    2007-01-01

    of pectins. The mode of action of BcPeh28A on different substrates has been investigated and its enzymatic mechanism elucidated. The hydrolysis of polygalacturonate indicates that BcPeh28A is a non-processive enzyme that releases oligomers with chain lengths ranging from two to eight. By inspection...

  17. Biological Role of Pigment Production for the Bacterial Phytopathogen Pantoea stewartii subsp. stewartii

    OpenAIRE

    Mohammadi, Mojtaba; Burbank, Lindsey; Roper, M. Caroline

    2012-01-01

    Pantoea stewartii subsp. stewartii, the causal agent of Stewart's wilt of sweet corn, produces a yellow carotenoid pigment. A nonpigmented mutant was selected from a bank of mutants generated by random transposon mutagenesis. The transposon insertion site was mapped to the crtB gene, encoding a putative phytoene synthase, an enzyme involved in the early steps of carotenoid biosynthesis. We demonstrate here that the carotenoid pigment imparts protection against UV radiation and also contribute...

  18. Biological role of pigment production for the bacterial phytopathogen Pantoea stewartii subsp. stewartii.

    Science.gov (United States)

    Mohammadi, Mojtaba; Burbank, Lindsey; Roper, M Caroline

    2012-10-01

    Pantoea stewartii subsp. stewartii, the causal agent of Stewart's wilt of sweet corn, produces a yellow carotenoid pigment. A nonpigmented mutant was selected from a bank of mutants generated by random transposon mutagenesis. The transposon insertion site was mapped to the crtB gene, encoding a putative phytoene synthase, an enzyme involved in the early steps of carotenoid biosynthesis. We demonstrate here that the carotenoid pigment imparts protection against UV radiation and also contributes to the complete antioxidant pathway of P. stewartii. Moreover, production of this pigment is regulated by the EsaI/EsaR quorum-sensing system and significantly contributes to the virulence of the pathogen in planta.

  19. Comparative and functional genomic analyses of the pathogenicity of phytopathogen Xanthomonas campestris pv. campestris

    OpenAIRE

    Qian, Wei; Jia, Yantao; Ren, Shuang-Xi; He, Yong-Qiang; Feng, Jia-Xun; Lu, Ling-Feng; Sun, Qihong; Ying, Ge; Tang, Dong-Jie; Tang, Hua; Wu, Wei; Hao, Pei; Wang, Lifeng; Jiang, Bo-Le; Zeng, Shenyan

    2005-01-01

    Xanthomonas campestris pathovar campestris (Xcc) is the causative agent of crucifer black rot disease, which causes severe losses in agricultural yield world-wide. This bacterium is a model organism for studying plant-bacteria interactions. We sequenced the complete genome of Xcc 8004 (5,148,708 bp), which is highly conserved relative to that of Xcc ATCC 33913. Comparative genomics analysis indicated that, in addition to a significant genomic-scale rearrangement cross the replication axis bet...

  20. Comparative and functional genomic analyses of the pathogenicity of phytopathogen Xanthomonas campestris pv. campestris.

    Science.gov (United States)

    Qian, Wei; Jia, Yantao; Ren, Shuang-Xi; He, Yong-Qiang; Feng, Jia-Xun; Lu, Ling-Feng; Sun, Qihong; Ying, Ge; Tang, Dong-Jie; Tang, Hua; Wu, Wei; Hao, Pei; Wang, Lifeng; Jiang, Bo-Le; Zeng, Shenyan; Gu, Wen-Yi; Lu, Gang; Rong, Li; Tian, Yingchuan; Yao, Zhijian; Fu, Gang; Chen, Baoshan; Fang, Rongxiang; Qiang, Boqin; Chen, Zhu; Zhao, Guo-Ping; Tang, Ji-Liang; He, Chaozu

    2005-06-01

    Xanthomonas campestris pathovar campestris (Xcc) is the causative agent of crucifer black rot disease, which causes severe losses in agricultural yield world-wide. This bacterium is a model organism for studying plant-bacteria interactions. We sequenced the complete genome of Xcc 8004 (5,148,708 bp), which is highly conserved relative to that of Xcc ATCC 33913. Comparative genomics analysis indicated that, in addition to a significant genomic-scale rearrangement cross the replication axis between two IS1478 elements, loss and acquisition of blocks of genes, rather than point mutations, constitute the main genetic variation between the two Xcc strains. Screening of a high-density transposon insertional mutant library (16,512 clones) of Xcc 8004 against a host plant (Brassica oleraceae) identified 75 nonredundant, single-copy insertions in protein-coding sequences (CDSs) and intergenic regions. In addition to known virulence factors, full virulence was found to require several additional metabolic pathways and regulatory systems, such as fatty acid degradation, type IV secretion system, cell signaling, and amino acids and nucleotide metabolism. Among the identified pathogenicity-related genes, three of unknown function were found in Xcc 8004-specific chromosomal segments, revealing a direct correlation between genomic dynamics and Xcc virulence. The present combination of comparative and functional genomic analyses provides valuable information about the genetic basis of Xcc pathogenicity, which may offer novel insight toward the development of efficient methods for prevention of this important plant disease.

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

    Science.gov (United States)

    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. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Detection of double stranded RNA in phytopathogenic Macrophomina phaseolina causing charcoal rot in Cyamopsis tetragonoloba.

    Science.gov (United States)

    Arora, Pooja; Dilbaghi, Neeraj; Chaudhury, Ashok

    2012-03-01

    One hundred one isolates of Macrophomina phaseolina from various hosts and eco-geographical locations were employed for elucidating relationships among genetic diversity and virulence. Highly pathogenic, moderately pathogenic, and hypovirulent cluster bean specific isolates were identified. In order to correlate respective phenotypes of plant pathogenic fungus multiple and complex patterns of dsRNA elements were analyzed. Double-stranded ribonucleic acids (dsRNA) are ubiquitous in all major groups and most of them have vast potential as biological control agents for fungi. Rate of virulence and its further association could ascertain by host plant and their fungal genotypes. Variability of the fungal genotypes decides the link between the complexity of dsRNA with different variants and the change in virulence pattern. Double-stranded RNA was identified in approximately 21.7% of M. phaseolina isolates from charcoal rot infected cluster bean varieties. After recurrent laboratory transfer on culture media, the preponderance of the isolates harboring dsRNAs developed degenerate culture phenotypes and showed reduced virulence (hypovirulence) to cluster bean. Macrophomina has successfully showed diversified and reproducible banding profile in dsRNA containing/free isolates. This is the first report of hypovirulence and detection of dsRNA in Macrophomina phaseolina isolates of cluster bean origin.

  3. Optimization for rapid synthesis of silver nanoparticles and its effect on phytopathogenic fungi

    Science.gov (United States)

    Krishnaraj, C.; Ramachandran, R.; Mohan, K.; Kalaichelvan, P. T.

    In this present study, silver nanoparticles were synthesized by green chemistry approach using Acalypha indica leaf extract as reducing agents. The reaction medium employed in the synthesis process was optimized to attain better yield, controlled size and stability. Further, the biosynthesized silver nanoparticles were conformed through UV-vis spectrum, XRD and HR-TEM analyses. Different concentration of silver nanoparticles were tested to know the inhibitory effect of fungal plant pathogens namely Alternaria alternata, Sclerotinia sclerotiorum, Macrophomina phaseolina, Rhizoctonia solani, Botrytis cinerea and Curvularia lunata. Interestingly, 15 mg concentration of silver nanoparticles showed excellent inhibitory activity against all the tested pathogens. Thus, the obtained results clearly suggest that silver nanoparticles may have important applications in controlling various plant diseases caused by fungi.

  4. Purification and molecular characterization of a sialic acid specific lectin from the phytopathogenic fungus Macrophomina phaseolina.

    Science.gov (United States)

    Bhowal, Jayati; Guha, Arun Kumar; Chatterjee, Bishnu Pada

    2005-09-05

    A lectin was isolated and purified from the culture filtrate of the plant pathogenic fungus Macrophomina phaseolina by a combination of ammonium sulfate precipitation, affinity chromatography on fetuin-Sepharose 4B and ion-exchange chromatography on DEAE-A 50. The lectin designated MPL was homogeneous by PAGE and HPLC and a monomeric protein with a molecular weight of approximately 34 kDa as demonstrated by SDS-PAGE. It is a glycoprotein and agglutinated human erythrocytes regardless of the human blood type. Neuraminidase treatment of erythrocytes reduced the agglutination activity of the lectin. It is thermally stable and exhibits maximum activity between pH 6 and 7.2. Its carbohydrate binding specificity was investigated both by hapten inhibition of hemagglutination and by enzyme-conjugated lectin inhibition assay. Although, M. phaseolina lectin bound sialic acid, it exhibited binding affinity towards neuraminyl oligosaccharides of N-linked glycoproteins, alpha-Neu5Ac-(2-->3)-beta-Gal-(1-->4)-GlcNAc being maximum.

  5. Growth Behavior of Phytopathogen Clavibacter michiganensis ssp. sepedonicus Treated with Selenium Biocomposites of Mushroom Origin

    Directory of Open Access Journals (Sweden)

    A.I. Perfileva

    2016-02-01

    Full Text Available The results of studying the effect of biologically obtained selenium nanocomposites on the bacterium Clavibacter michiganensis ssp. sepedonicus (Cms are presented. Cms is a Gram-positive bacterium, which causes one of the most dangerous potato diseases, ring rot. The effective alongside ecologically safe methods for combating Cms are lacking. As the agents feasible for use in this purpose, we examined the selenium nanocomposites obtained from the macrobasidiomycetes' submerged cultures. For exploring the bionanocomposites effect on Cms, the methods of agar well diffusion, the suspension turbidity measurement, and the colony-forming units count were applied. The results showed that all the nanocomposites under study lowered the bacterial suspension's absorption values compared to the reference specimen, that testified to the observation of bacteriostatic effect of the agents tested. The suppression action of nanocompostes was elucidated by means of both agar well diffusion assay and colony-forming units count. Thus, the results obtained demonstrate the occurrence of bacteriostatic and bactericidal effects of the substances under study, and favor the supposition on advisability of further research into the selenium nanocomposites as the agents for agricultural recovery from the bacterial pathogens.

  6. Characterization of the plasmid encoded virulence region pat-1 of phytopathogenic Clavibacter michiganensis subsp. michiganensis.

    Science.gov (United States)

    Dreier, J; Meletzus, D; Eichenlaub, R

    1997-03-01

    The tomato pathogen Clavibacter michiganensis subsp. michiganensis NCPPB382, causing bacterial wilt and canker, harbors two plasmids, pCM1 (27.5 kb) and pCM2 (72 kb), carrying genes involved in virulence. The region of plasmid pCM2 encoding the pathogenicity locus pat-1 was mapped by deletion analysis and complementation studies to a 1.5-kb Bg/II/SmaI DNA fragment. Introduction of the pat-1 region into endophytic, plasmid-free isolates of C. michiganensis subsp. michiganensis converted these bacteria into virulent pathogens. Based on the nucleotide sequence of the pat-1 region, an open reading frame (ORF1) can be predicted, coding for a protein of 280 amino acids and 29.7 kDa with homology to serine proteases. Introduction of a frame-shift mutation in ORF1 leads to a loss of the pathogenic phenotype. Northern (RNA) hybridizations identified an 1.5-knt transcript of the pat-1 structural gene. The site of transcription initiation was mapped by primer extension and a typical -10/-35 region was located with significant homology to the consensus Escherichia coli sigma 70 and Bacillus subtilis sigma 43 promoters. Downstream of the pat-1 structural gene, a peculiar repetitive sequence motif (pat-1rep) is located, consisting of 20 direct tandem repeats preceded by a run of 14 guanosine residues. DNA sequences homologous to pat-1rep were isolated and characterized from four virulent C. michiganensis subsp. michiganensis strains exhibiting a high extent of structural conservation. The deletion of this repetitive sequence reduced virulence significantly but did not lead to a complete loss of the virulence phenotype.

  7. A TaqMan-PCR protocol for quantification and differentiation of the phytopathogenic Clavibacter michiganensis subspecies.

    Science.gov (United States)

    Bach, H-J; Jessen, I; Schloter, M; Munch, J C

    2003-01-01

    Real-time TaqMan-PCR assays were developed for detection, differentiation and absolute quantification of the pathogenic subspecies of Clavibacter michiganensis (Cm) in one single PCR run. The designed primer pair, targeting intergenic sequences of the rRNA operon (ITS) common in all subspecies, was suitable for the amplification of the expected 223-nt DNA fragments of all subspecies. Closely related bacteria were completely discriminated, except of Rathayibacter iranicus, from which weak PCR product bands appeared on agarose gel after 35 PCR cycles. Sufficient specificity of PCR detection was reached by introduction of the additional subspecies specific probes used in TaqMan-PCR. Only Cm species were detected and there was clear differentiation among the subspecies C. michiganensis sepedonicus (Cms), C. michiganensis michiganensis (Cmm), C. michiganensis nebraskensis (Cmn), C. michiganensis insidiosus (Cmi) and C. michiganensis tessellarius (Cmt). The TaqMan assays were optimized to enable a simultaneous quantification of each subspecies. Validity is shown by comparison with cell counts.

  8. Antifungal Depsidone Metabolites from Cordyceps dipterigena, an Endophytic Fungus Antagonistic to the Phytopathogen Gibberella fujikuroi

    Science.gov (United States)

    Varughese, Titto; Riosa, Nivia; Higginbotham, Sarah; Arnold, A. Elizabeth; Coley, Phyllis D.; Kursar, Thomas A.; Gerwick, William H.; Cubilla Rios, L.

    2012-01-01

    Among thirty four endophytic fungal strains screened for in vitro antagonism, the endophytic fungus Cordyceps dipterigena was found to strongly inhibit mycelial growth of the plant pathogenic fungus Gibberella fujikuroi. Two new depsidone metabolites, cordycepsidone A (1) and cordycepsidone B (2), were isolated from the PDA culture extract of C. dipterigena and identified as being responsible for the antifungal activity. Elucidation of their chemical structures was carried out using 1D and 2D NMR spectroscopy in combination with IR and MS spectroscopic data. Cordycepsidone A displayed strong and dose-dependent antifungal activity against the plant pathogenic fungus Gibberella fujikuroi. The isolates were inactive in bioassays for malaria (Plasmodium falciparum), leishmaniasis (Leishmania donovani), Chagas’s disease (Trypanosoma cruzi), and cytotoxicity at 10 μg/mL. The compounds were also found to be inactive against several bacterial strains at 50 μg/mL. PMID:22707798

  9. Antagonist capacity of Newly Isolated Strains of Pseudomonas Fluorescens against Three Important Phytopathogenic Bacteria

    Directory of Open Access Journals (Sweden)

    Reynaldo D.L. Cruz-Quiroz

    2011-01-01

    Full Text Available Problem statement: Phytopatogenic bacteria cause several damages to plants with important economical consequences. They provoke losses of product quality affecting all commercial chain of crops, for this reason, their control is a priority. Approach: We evaluated antagonist capacity of newly isolated Pseudonomas fluorescens strains against three important phytopatogenic bacteria (Clavibacter michiganensis, Xanthomonas axonopodis and Erwinia carotovora. Soils from commercial cropping of Capsicum annum L of several Mexican regions were used to isolate P. fluorescens strains. Results: Isolates producing flourescein were purified on King B agar and biochemically identified. Crude extracts with and without cells were produced in King B broths and their antagonist capacities were evaluated by the plate diffusion procedure on nutritive agar. Conclusion: Obtained results demonstrated that cell free extracts exhibited a limited antagonist capacity in comparison of those extracts with cells, which showed an excellent capacity to inhibit the growth of C. michiganensis, X. axonopodis and E. carotovora, demonstrating the intracellular nature of the bioactive metabolites associated to bacterial growth inhibition.

  10. Studies on phytopathogenic and saprotrophic fungi in rush associations of Lake Glinno (NW Poland

    Directory of Open Access Journals (Sweden)

    Kinga Mazurkiewicz-Zapałowicz

    2013-12-01

    Full Text Available During the vegetation seasons in years 2004-2005 the health state of rush plant species from Phragmition and Magnocaricion alliances around the Lake Glinno was investigated. From 13 plant species with disease symptoms 94 species of fungi and FLO were isolated. The highest mycological biodiversity was stated in Phragmitetum australis (24 species and Thelypteridi-Phragmitetum (27 species plant associations. The host species in which the biggest number of fungi and FLO species was observed were: Phragmites australis (37 species and Carex acutiformis (25 species. The highest mycological similarity based on the Jaccard-Sörensen coefficient occurred between Caricetum acutiformis and Glycerietum maximae plant associations (50% whereas the lowest value of the coefficient represented Glycerietum maximae and Phalaridetum arundinaceae associations (7%.

  11. Exploring Western Ghats microbial diversity for antagonistic microorganisms against fungal phytopathogens of pepper and chickpea

    Directory of Open Access Journals (Sweden)

    B.N. RAMKUMAR

    2015-08-01

    Full Text Available Newly isolated microbial cultures from Western Ghat soil samples of Kerala region in India were screened for antagonistic activity by well diffusion and dual culture plating against Phytophthora capsici and Rhizoctonia solani, infecting pepper and chickpea, respectively. Bioactive samples were made by varying solvent extraction of the culture broths of the potent isolates belongs to Actinomycetes, Pseudomonas, Bacillus and Trichoderma. The efficacy of the isolates to produce other potent antifungal metabolites such as cell wall degrading enzymes, HCN and volatile compounds were also checked. Treatment with antagonistic isolates in vivo under greenhouse conditions revealed significant reduction of the disease intensity of foot rot disease of black pepper and collar rot of chick pea.

  12. Ancestral state reconstruction infers phytopathogenic origins of sooty blotch and flyspeck fungi on apple.

    Science.gov (United States)

    Ismail, Siti Izera; Batzer, Jean Carlson; Harrington, Thomas C; Crous, Pedro W; Lavrov, Dennis V; Li, Huanyu; Gleason, Mark L

    2016-01-01

    Members of the sooty blotch and flyspeck (SBFS) complex are epiphytic fungi in the Ascomycota that cause economically damaging blemishes of apples worldwide. SBFS fungi are polyphyletic, but approx. 96% of SBFS species are in the Capnodiales. Evolutionary origins of SBFS fungi remain unclear, so we attempted to infer their origins by means of ancestral state reconstruction on a phylogenetic tree built utilizing genes for the nuc 28S rDNA (approx. 830 bp from near the 59 end) and the second largest subunit of RNA polymerase II (RPB2). The analyzed taxa included the well-known genera of SBFS as well as non-SBFS fungi from seven families within the Capnodiales. The non-SBFS taxa were selected based on their distinct ecological niches, including plant-parasitic and saprophytic species. The phylogenetic analyses revealed that most SBFS species in the Capnodiales are closely related to plant-parasitic fungi. Ancestral state reconstruction provided strong evidence that plant-parasitic fungi were the ancestors of the major SBFS lineages. Knowledge gained from this study may help to better understand the ecology and evolution of epiphytic fungi. © 2016 by The Mycological Society of America.

  13. Leaf-cutting ant fungi produce cell wall degrading pectinase complexes reminiscent of phytopathogenic fungi

    DEFF Research Database (Denmark)

    Schiøtt, Morten; Rogowska-Wrzesinska, Adelina; Roepstorff, Peter

    2010-01-01

    Leaf-cutting (attine) ants use their own fecal material to manure fungus gardens, which consist of leaf material overgrown by hyphal threads of the basidiomycete fungus Leucocoprinus gongylophorus that lives in symbiosis with the ants. Previous studies have suggested that the fecal droplets conta...

  14. Whole genome sequences and annotation of Micrococcus luteus SUBG006, a novel phytopathogen of mango

    Directory of Open Access Journals (Sweden)

    Purvi M. Rakhashiya

    2015-12-01

    Full Text Available Actinobaceria, Micrococcus luteus SUBG006 was isolated from infected leaves of Mangifera indica L. vr. Nylon in Rajkot, (22.30°N, 70.78°E, Gujarat, India. The genome size is 3.86 Mb with G + C content of 69.80% and contains 112 rRNA sequences (5S, 16S and 23S. The whole genome sequencing has been deposited in DDBJ/EMBL/GenBank under the accession number JOKP00000000.

  15. Development of Conductive Polymer Analysis for the Rapid Detection and Identification of Phytopathogenic Microbes

    Science.gov (United States)

    A. Dan Wilson; D.G. Lester; C.S. Oberle

    2004-01-01

    Conductive polymer analysis, a type of electronic aroma detection technology, was evaluated for its efficacy in the detection, identification, and discrimination of plant-pathogenic microorganisms on standardized media and in diseased plant tissues. The method is based on the acquisition of a diagnostic electronic fingerprint derived from multisensor responses to...

  16. Characterization of a new aryl-alcohol oxidase secreted by the phytopathogenic fungus Ustilago maydis.

    Science.gov (United States)

    Couturier, Marie; Mathieu, Yann; Li, Ai; Navarro, David; Drula, Elodie; Haon, Mireille; Grisel, Sacha; Ludwig, Roland; Berrin, Jean-Guy

    2016-01-01

    The discovery of novel fungal lignocellulolytic enzymes is essential to improve the breakdown of plant biomass for the production of second-generation biofuels or biobased materials in green biorefineries. We previously reported that Ustilago maydis grown on maize secreted a diverse set of lignocellulose-acting enzymes including hemicellulases and putative oxidoreductases. One of the most abundant proteins of the secretome was a putative glucose-methanol-choline (GMC) oxidoreductase. The phylogenetic prediction of its function was hampered by the few characterized members within its clade. Therefore, we cloned the gene and produced the recombinant protein to high yield in Pichia pastoris. Functional screening using a library of substrates revealed that this enzyme was able to oxidize several aromatic alcohols. Of the tested aryl-alcohols, the highest oxidation rate was obtained with 4-anisyl alcohol. Oxygen, 1,4-benzoquinone, and 2,6-dichloroindophenol can serve as electron acceptors. This GMC oxidoreductase displays the characteristics of an aryl-alcohol oxidase (E.C.1.1.3.7), which is suggested to act on the lignin fraction in biomass.

  17. Model analysis for plant disease dynamics co-mediated by herbivory and herbivore-borne phytopathogens.

    Science.gov (United States)

    Nakazawa, Takefumi; Yamanaka, Takehiko; Urano, Satoru

    2012-08-23

    Plants are subject to diseases caused by pathogens, many of which are transmitted by herbivorous arthropod vectors. To understand plant disease dynamics, we studied a minimum hybrid model combining consumer-resource (herbivore-plant) and susceptible-infected models, in which the disease is transmitted bi-directionally between the consumer and the resource from the infected to susceptible classes. Model analysis showed that: (i) the disease is more likely to persist when the herbivore feeds on the susceptible plants rather than the infected plants, and (ii) alternative stable states can exist in which the system converges to either a disease-free or an endemic state, depending on the initial conditions. The second finding is particularly important because it suggests that the disease may persist once established, even though the initial prevalence is low (i.e. the R(0) rule does not always hold). This situation is likely to occur when the infection improves the plant nutritive quality, and the herbivore preferentially feeds on the infected resource (i.e. indirect vector-pathogen mutualism). Our results highlight the importance of the eco-epidemiological perspective that integration of tripartite interactions among host plant, plant pathogen and herbivore vector is crucial for the successful control of plant diseases.

  18. Association between virulence and triazole tolerance in the phytopathogenic fungus Mycosphaerella graminicola.

    Directory of Open Access Journals (Sweden)

    Lina Yang

    Full Text Available Host resistance and synthetic antimicrobials such as fungicides are two of the main approaches used to control plant diseases in conventional agriculture. Although pathogens often evolve to overcome host resistance and antimicrobials, the majority of reports have involved qualitative host - pathogen interactions or antimicrobials targeting a single pathogen protein or metabolic pathway. Studies that consider jointly the evolution of virulence, defined as the degree of damage caused to a host by parasite infection, and antimicrobial resistance are rare. Here we compared virulence and fungicide tolerance in the fungal pathogen Mycosphaerella graminicola sampled from wheat fields across three continents and found a positive correlation between virulence and tolerance to a triazole fungicide. We also found that quantitative host resistance selected for higher pathogen virulence. The possible mechanisms responsible for these observations and their consequences for sustainable disease management are discussed.

  19. Mycoparasitism studies of Trichoderma species against three phytopathogenic fungi: evaluation of antagonism and hydrolytic enzyme production.

    Science.gov (United States)

    Qualhato, Thiago Fernandes; Lopes, Fabyano Alvares Cardoso; Steindorff, Andrei Stecca; Brandão, Renata Silva; Jesuino, Rosália Santos Amorim; Ulhoa, Cirano José

    2013-09-01

    Trichoderma spp. are used for biocontrol of several plant pathogens. However, their efficient interaction with the host needs to be accompanied by production of secondary metabolites and cell wall-degrading enzymes. Three parameters were evaluated after interaction between four Trichoderma species and plant-pathogenic fungi: Fusarium solani, Rhizoctonia solani and Sclerotinia sclerotiorum. Trichoderma harzianum and T. asperellum were the most effective antagonists against the pathogens. Most of the Trichoderma species produced toxic volatile metabolites, having significant effects on growth and development of the plant pathogens. When these species were grown in liquid cultures with cell walls from these plant pathogens, they produced and secreted β-1,3-glucanase, NAGAse, chitinase, acid phosphatase, acid proteases and alginate lyase.

  20. The effect of polyamine biosynthesis inhibition on growth and differentiation of the phytopathogenic fungus Sclerotinia sclerotiorum.

    Science.gov (United States)

    Pieckenstain, F L; Gárriz, A; Chornomaz, E M; Sánchez, D H; Ruiz, O A

    2001-12-01

    We studied the effects of several polyamine biosynthesis inhibitors on growth, differentiation, free polyamine levels and in vivo and in vitro activity of polyamine biosynthesis enzymes in Sclerotinia sclerotiorum. Alpha-Difluoromethylornithine (DFMO) and alpha-difluoromethylarginine (DFMA) were potent inhibitors of mycelial growth. The effect of DFMO was due to inhibition of ornithine decarboxylase (ODC). No evidence for the existence of an arginine decarboxylase (ADC) pathway was found. The effect of DFMA was partly due to inhibition of ODC, presumably after its conversion into DFMO by mycelial arginase, as suggested by the high activity of this enzyme detected both in intact mycelium and mycelial extracts. In addition, toxic effects of DFMA on cellular processes other than polyamine metabolism might have occurred. Cyclohexylamine (CHA) slightly inhibited mycelial growth and caused an important decrease of free spermidine associated with a drastic increase of free putrescine concentration. Methylglyoxal bis-[guanyl hydrazone] (MGBG) had no effect on mycelial growth. Excepting MGBG, all the inhibitors strongly decreased sclerotial formation. Results demonstrate that sclerotial development is much more sensitive to polyamine biosynthesis inhibition than mycelial growth. Our results suggest that mycelial growth can be supported either by spermidine or putrescine, while spermidine (or the putrescine/spermidine ratio) is important for sclerotial formation to occur. Ascospore germination was completely insensitive to the inhibitors.

  1. Identification and Pathogenicity of Phytopathogenic Bacteria Associated with Soft Rot Disease of Girasole Tuber

    OpenAIRE

    Mamdoh Ewis ISMAIL; Abdel-Monaim, Montaser Fawzy; Yasser Mahmoud MOSTAFA

    2012-01-01

    During 2010-2011 growing seasons six bacterial isolates were separated from naturally infected girasole plants tubers (Helianthus tuberosus L.) cv. �Balady�, showing soft rot, collected from experimental Farm of the Faculty of Agriculture, in El-Minia University, Egypt. Pathogenicity tests showed various virulence for the bacteria isolated from girasole tubers, found pathogenic. These organisms were characterized as rod-shaped, Gram negative, ?-methyl-d-glucoside medium, reducing substances f...

  2. Identification and Pathogenicity of Phytopathogenic Bacteria Associated with Soft Rot Disease of Girasole Tuber

    Directory of Open Access Journals (Sweden)

    Mamdoh Ewis ISMAIL

    2012-02-01

    Full Text Available During 2010-2011 growing seasons six bacterial isolates were separated from naturally infected girasole plants tubers (Helianthus tuberosus L. cv. Balady, showing soft rot, collected from experimental Farm of the Faculty of Agriculture, in El-Minia University, Egypt. Pathogenicity tests showed various virulence for the bacteria isolated from girasole tubers, found pathogenic. These organisms were characterized as rod-shaped, Gram negative, ?-methyl-d-glucoside medium, reducing substances from sucrose, phos, phatase activity and deep cavities on pectate medium. Otherwise, diagnostic tests suggested that the pathogen was Erwinia carotovora ssp. carotovora. The isolated bacteria caused soft rot of wounded tubers when inoculated into tissues. The bacterial isolates were compared for their degree of pathogenicity as well as for differences in specific symptoms, induced in different hosts. The tested isolates could infect several host ranges, such as fruits of apricot, apple, olive, lemon, squash, eggplant and potato tubers, bulbs and garlic and onion cloves, roots radish, carrot, sweet potato and rape. On the other hand, no symptoms were exhibited on pods of bean and cowpea, faba bean, fruits of pepper and tomato. The extracts of experimentally diseased girasole tubers were active in pectinase and also in caboxymethyl cellulose at pH 6 compared to enzyme activities in healthy tissues. Also, the isolated bacteria increased the total and reducing sugars in infected tissues.

  3. Phylogeography and population structure of the biologically invasive phytopathogen Erwinia amylovora inferred using minisatellites.

    Science.gov (United States)

    Bühlmann, Andreas; Dreo, Tanja; Rezzonico, Fabio; Pothier, Joël F; Smits, Theo H M; Ravnikar, Maja; Frey, Jürg E; Duffy, Brion

    2014-07-01

    Erwinia amylovora causes a major disease of pome fruit trees worldwide, and is regulated as a quarantine organism in many countries. While some diversity of isolates has been observed, molecular epidemiology of this bacterium is hindered by a lack of simple molecular typing techniques with sufficiently high resolution. We report a molecular typing system of E. amylovora based on variable number of tandem repeats (VNTR) analysis. Repeats in the E. amylovora genome were identified with comparative genomic tools, and VNTR markers were developed and validated. A Multiple-Locus VNTR Analysis (MLVA) was applied to E. amylovora isolates from bacterial collections representing global and regional distribution of the pathogen. Based on six repeats, MLVA allowed the distinction of 227 haplotypes among a collection of 833 isolates of worldwide origin. Three geographically separated groups were recognized among global isolates using Bayesian clustering methods. Analysis of regional outbreaks confirmed presence of diverse haplotypes but also high representation of certain haplotypes during outbreaks. MLVA analysis is a practical method for epidemiological studies of E. amylovora, identifying previously unresolved population structure within outbreaks. Knowledge of such structure can increase our understanding on how plant diseases emerge and spread over a given geographical region.

  4. Huitlacoche (corn smut), caused by the phytopathogenic fungus Ustilago maydis, as a functional food.

    Science.gov (United States)

    Juárez-Montiel, Margarita; Ruiloba de León, Sandra; Chávez-Camarillo, Griselda; Hernández-Rodríguez, César; Villa-Tanaca, Lourdes

    2011-01-01

    In recent years the need has arisen to study and develop (or re-discover) foods that have nutritional characteristics as well as specific functions, such as improving health and/or reducing the risk of disease. For this reason knowledge of the nutritional value of food is important to promote greater consumer acceptance. In Mexico huitlacoche (also, cuitlacoche) has traditionally been prized as a delicacy since the time of the Aztecs and is currently being studied as a potential functional food and as a producer of natural bioactive substances that are used in fortifying foods. To present an updated review about the properties of the huitlacoche (corn smut) as functional food. A bibliographic search was performed and data were discussed. The data of the works reviewed here show that huitlacoche contains many compounds that confer to it unique organoleptic and nutraceutical characteristics. The content of bioactive substances in huitlacoche supports the proposal that this is a good functional food as well as producer of compounds to enrich other foods. Copyright © 2010 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  5. Genome sequences of three phytopathogenic species of the Magnaporthaceae family of fungi

    Science.gov (United States)

    Magnaporthaceae is a family of ascomycetes that includes three fungi of great economic importance: Magnaporthe oryzae, Gaeumannomyces graminis var. tritici, and Magnaporthe poae. These three fungi cause widespread disease and loss in cereal and grass crops, including rice blast disease (M. oryzae), ...

  6. Tri6 is a global transcription regulator in the phytopathogen Fusarium graminearum.

    Directory of Open Access Journals (Sweden)

    Charles G Nasmith

    2011-09-01

    Full Text Available In F. graminearum, the transcriptional regulator Tri6 is encoded within the trichothecene gene cluster and regulates genes involved in the biosynthesis of the secondary metabolite deoxynivalenol (DON. The Tri6 protein with its Cys₂His₂ zinc-finger may also conform to the class of global transcription regulators. This class of global transcriptional regulators mediate various environmental cues and generally responds to the demands of cellular metabolism. To address this issue directly, we sought to find gene targets of Tri6 in F. graminearum grown in optimal nutrient conditions. Chromatin immunoprecipitation followed by Illumina sequencing (ChIP-Seq revealed that in addition to identifying six genes within the trichothecene gene cluster, Tri1, Tri3, Tri6, Tri7, Tri12 and Tri14, the ChIP-Seq also identified 192 additional targets potentially regulated by Tri6. Functional classification revealed that, among the annotated genes, ∼40% are associated with cellular metabolism and transport and the rest of the target genes fall into the category of signal transduction and gene expression regulation. ChIP-Seq data also revealed Tri6 has the highest affinity toward its own promoter, suggesting that this gene could be subject to self-regulation. Electro mobility shift assays (EMSA performed on the promoter of Tri6 with purified Tri6 protein identified a minimum binding motif of GTGA repeats as a consensus sequence. Finally, expression profiling of F. graminearum grown under nitrogen-limiting conditions revealed that 49 out of 198 target genes are differentially regulated by Tri6. The identification of potential new targets together with deciphering novel binding sites for Tri6, casts new light into the role of this transcriptional regulator in the overall growth and development of F. graminearum.

  7. Isolation and fusion of protoplasts from the phytopathogenic fungus Sclerotium rolfsii (Sacc.

    Directory of Open Access Journals (Sweden)

    Sikandar Hayat

    2010-03-01

    Full Text Available Sclerotium rolfsii (Sacc. is a serious plant pathogenic fungus and lacks perfect (basidial stage in production. Protoplast fusion technology was employed to reconstruct fusants from this fungus. Two strains designated as A and R were used. Maximum protoplast yields of 3.8x10(5 /g mycelia and 2.8x10(5 /g mycelia were formed in strains A and R respectively. Osmotic stabilizer sucrose 1M gave maximum yield. Lysing enzyme at the rate of 15mg/ml was found best for yield. Fusion of protoplasts from strains A and R was carried out in fusion media containing PEG 4000 30% (w/v with 0.2mM CaCl2. Four fusants F1, F2, F3 and F4 were recovered. Morphological, physiological and pathogenic characters of fusants were compared with parent strains on carrots, beans and tomato.

  8. Induction of infection in Teosinte (Zea diploperennis through the phytopathogen Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Martha Concepción Pérez Díaz

    2017-01-01

    Full Text Available Introduction: The corn and teosinte share morphological and molecular similarities latter being those that support the theory of teosinte (Zea parviglumis as its predecessor, both species are attacked by specific pathogens like Ustilago maydis. Objective: To analyze the infectious process that presents U. maydis on the variety of the teosinte Zea diploperennis. Materials and Methods: We used the strain of U. maydis FB-D12, which was kept a culture media rich in nutrients (CPES pH 7.0. Viable cells without morphological alterations to the inoculation method of puncture in teosinte seedlings were used. Monitoring of infection was carried out every 24 hours by measuring concentration of chlorophyll and plant tissue through microscopic observation Results: In the seedlings of Zea diploperennis inoculated with U. maydis the symptoms of the infection were presented, wilt and chlorosis in the leaves; The chlorosis was confirmed with the low concentration of chlorophyll 12 days later to the inoculation. In the microscopic observation of cuts of the tissue plant was found mycelium long and branched from the third day of the inoculation, until the appearance of tumors in seedlings of 45 days. Conclusions: The typical signs of infection with Ustilago maydis in the variety of teosinte Zea diploperennis do not differ from those reported for corn. Ustilago maydis presents its full life cycle within the plant, confirming that the diploperennis variety is susceptible.

  9. Features of isolation and identification of phytopathogenic bacteria and search of their natural antagonists

    Directory of Open Access Journals (Sweden)

    O. S. Stonchyus

    2010-08-01

    Full Text Available Microflora of vegetables affected by bacteriosis was isolated and investigated. On the basis of morphological, cultural, physiological and biochemical characteristics of selected strains they were identified, and their plants pathogenicity was shown. The antagonistic influence of strain Bacillus thuringiensis var. thuringiensis IMV B-7186 on isolated bacterial culture was studied.

  10. Antibacterial activity of carob (Ceratonia siliqua L.) extracts against phytopathogenic bacteria Pectobacterium atrosepticum.

    Science.gov (United States)

    Meziani, Saïda; Oomah, B Dave; Zaidi, Farid; Simon-Levert, Annabel; Bertrand, Cédric; Zaidi-Yahiaoui, Rachida

    2015-01-01

    Acetone and ethanol extracts of carob (Ceratonia siliqua L.) leaf and pods were evaluated for their in vitro inhibitory ability against the pectinolytic Gram negative Pectobacterium atrosepticum (Pca, CFBP-5384) bacteria, the causal agent of potato soft rot. Potato (Solanum tuberosum, var nicola) tuber rot tissues obtained after 5 day bacterial inoculation was analyzed by LC-MS and GC-MS to study Pca pathogenicity. Trans/cis N-feruloylputrescine was identified in potato tuber after 5-day inoculation with Pca in a dark moist chamber. Although glycoalkoloid (α-chaconine and α-solanine) production increased due to Pca soft rot infection, it was not a resistance-determining factor. Many secondary metabolites were identified including the phytoalexins solavetivone and fatty acids responsible for plant defence responses. Acetone extract of carob leaf (FCA) exhibited the strongest inhibitory effect (IC50 = 1.5 mg/ml) and displayed synergistic antimicrobial effect in the presence of infected potato tuber extract (Pdt-Pca extract) against Pca. This synergy could be used in an integrated control program against potato soft rot pathogens, thereby reducing chemical treatments.

  11. Exploring Western Ghats microbial diversity for antagonistic microorganisms against fungal phytopathogens of pepper and chickpea

    National Research Council Canada - National Science Library

    B.N. RAMKUMAR; K.M. NAMPOOTHIRI; U. SHEEBA; P. JAYACHANDRAN; N.S. SREESHMA; S.M. SNEHA; K.S. Meenakumari; P. Sivaprasad

    2015-01-01

    ... capsici and Rhizoctonia solani, infecting pepper and chickpea, respectively. Bioactive samples were made by varying solvent extraction of the culture broths of the potent isolates belongs to Actinomycetes, Pseudomonas, Bacillus and Trichoderma...

  12. Ancestral state reconstruction infers phytopathogenic origins of sooty blotch and flyspeck fungi on apple

    NARCIS (Netherlands)

    Ismail, Siti Izera; Batzer, Jean Carlson; Harrington, Thomas C.; Crous, Pedro W.; Lavrov, Dennis V.; Li, Huanyu; Gleason, Mark L.

    2016-01-01

    Members of the sooty blotch and flyspeck (SBFS) complex are epiphytic fungi in the Ascomycota that cause economically damaging blemishes of apples worldwide. SBFS fungi are polyphyletic, but approx. 96% of SBFS species are in the Capnodiales. Evolutionary origins of SBFS fungi remain unclear, so we

  13. Ancestral state reconstruction infers phytopathogenic origins of sooty blotch and flyspeck fungi on apple

    NARCIS (Netherlands)

    Ismail, Siti Izera; Batzer, Jean Carlson; Harrington, Thomas C.; Crous, Pedro W.; Lavrov, Dennis V.; Li, Huanyu; Gleason, Mark L.

    2016-01-01

    Members of the sooty blotch and flyspeck (SBFS) complex are epiphytic fungi in the Ascomycota that cause economically damaging blemishes of apples worldwide. SBFS fungi are polyphyletic, but approx. 96% of SBFS species are in the Capnodiales. Evolutionary origins of SBFS fungi remain unclear, so

  14. Chitosan effects on phytopathogenic fungi and seed germination of Jatropha curcas L.

    Directory of Open Access Journals (Sweden)

    Diana Pabón-Baquero

    2015-01-01

    Full Text Available Jatropha curcas es una planta con gran potencial agrícola e industrial. En este estudio se aislaron dos hongos de semillas no germinadas. Los aislamientos fúngicos se identificaron morfológica y molecularmente como Fusarium equiseti y Curvularia lunata. Los efectos del quitosano se evaluaron sobre el crecimiento micelial, esporulación y germinación de esporas de F. equiseti y C. lunata. Además, se estudió el efecto sobre la germinación de las semillas de J. curcas. Los resultados demostraron que todas las concentraciones probadas de quitosano (0.5, 1.0, 2.0 y 4.0 mg·mL-1 inhibieron el crecimiento micelial de los hongos. Las respuestas de esporulación y germinación de esporas fueron diferentes dependiendo de la especie fúngica; el quitosano inhibió completamente la esporulación C. lunata y la germinación de esporas de F. equiseti. La inoculación con F. equiseti y C. lunata redujo la germinación de semillas de J. curcas 20 y 26.6 %, respectivamente; sin embargo, la aplicación de quitosano antes de la inoculación inhibió la actividad patogénica. En conclusión, el quitosano no afectó la germinación de las semillas y causó efectos inhibitorios en F. equiseti y C. lunata. Este es el primer reporte del efecto del quitosano en J. curcas.

  15. Genome structure and reproductive behaviour influence the evolutionary potential of a fungal phytopathogen.

    Directory of Open Access Journals (Sweden)

    Guillaume Daverdin

    Full Text Available Modern agriculture favours the selection and spread of novel plant diseases. Furthermore, crop genetic resistance against pathogens is often rendered ineffective within a few years of its commercial deployment. Leptosphaeria maculans, the cause of phoma stem canker of oilseed rape, develops gene-for-gene interactions with its host plant, and has a high evolutionary potential to render ineffective novel sources of resistance in crops. Here, we established a four-year field experiment to monitor the evolution of populations confronted with the newly released Rlm7 resistance and to investigate the nature of the mutations responsible for virulence against Rlm7. A total of 2551 fungal isolates were collected from experimental crops of a Rlm7 cultivar or a cultivar without Rlm7. All isolates were phenotyped for virulence and a subset was genotyped with neutral genetic markers. Virulent isolates were investigated for molecular events at the AvrLm4-7 locus. Whilst virulent isolates were not found in neighbouring crops, their frequency had reached 36% in the experimental field after four years. An extreme diversity of independent molecular events leading to virulence was identified in populations, with large-scale Repeat Induced Point mutations or complete deletion of AvrLm4-7 being the most frequent. Our data suggest that increased mutability of fungal genes involved in the interactions with plants is directly related to their genomic environment and reproductive system. Thus, rapid allelic diversification of avirulence genes can be generated in L. maculans populations in a single field provided that large population sizes and sexual reproduction are favoured by agricultural practices.

  16. Differential responses of Oryza sativa secondary metabolism to biotic interactions with cooperative, commensal and phytopathogenic bacteria.

    Science.gov (United States)

    Chamam, Amel; Wisniewski-Dyé, Florence; Comte, Gilles; Bertrand, Cédric; Prigent-Combaret, Claire

    2015-12-01

    Profiling of plant secondary metabolite allows to differentiate the different types of ecological interactions established between rice and bacteria. Rice responds to ecologically distinct bacteria by altering its content of flavonoids and hydroxycinnamic acid derivatives. Plants' growth and physiology are strongly influenced by the biotic interactions that plants establish with soil bacterial populations. Plants are able to sense and to respond accordingly to ecologically distinct bacteria, by inducing defense pathways against pathogens to prevent parasitic interactions, and by stimulating the growth of root-associated beneficial or commensal bacteria through root exudation. Plant secondary metabolism is expected to play a major role in this control. However, secondary metabolite responses of a same plant to cooperative, commensal and deleterious bacteria have so far never been compared. The impact of the plant growth-promoting rhizobacteria (PGPR) Azospirillum lipoferum 4B on the secondary metabolite profiles of two Oryza sativa L. cultivars (Cigalon and Nipponbare) was compared to that of a rice pathogen Burkholderia glumae AU6208, the causing agent of bacterial panicle blight and of a commensal environmental bacteria Escherichia coli B6. Root and shoot rice extracts were analyzed by reversed-phase high-performance liquid chromatography (RP-HPLC). Principal component analyses (PCAs) pinpointed discriminant secondary metabolites, which were characterized by mass spectrometry. Direct comparison of metabolic profiles evidenced that each bacterial ecological interaction induced distinct qualitative and quantitative modifications of rice secondary metabolism, by altering the content of numerous flavonoid compounds and hydroxycinnamic acid (HCA) derivatives. Secondary metabolism varied according to the cultivars and the interaction types, demonstrating the relevance of secondary metabolic profiling for studying plant-bacteria biotic interactions.

  17. The major Cu,Zn SOD of the phytopathogen Claviceps purpurea is not essential for pathogenicity

    NARCIS (Netherlands)

    Moore, S; De Vries, OMH; Tudzynski, P

    2002-01-01

    Superoxide dismutase (SOD) activities of the biotrophic pathogen Claviceps purpurea, which causes the ergot disease on a wide range of host grasses, were examined in axenic and pathogenic cultures. Almost all SOD activity in axenic culture originated from a single Cu,Zn SOD; a substantial part of th

  18. Agrobacterium tumefaciens-mediated genetic transformation of the phytopathogenic fungus Penicillium digitatum

    Institute of Scientific and Technical Information of China (English)

    Ji-ye WANG; Hong-ye LI

    2008-01-01

    Agrobacterium tumefaciens-mediated transformation (ATMT) system was assessed for conducting insertional mutagenesis in Penicillium digitatum, a major fungal pathogen infecting post-harvest citrus fruits. A transformation efficiency of up to 60 transformants per 106 conidia was achieved by this system. The integration of the hph gene into the fungal genome was verified by polymerase chain reaction (PCR) amplification and sequencing. These transformants tested were also shown to be mitotically stable. Southern blot analysis of 14 randomly selected transformants showed that the hph gene was randomly integrated as single copy into the fungal genome of P. digitatum. Thus, we conclude that ATMT of P. digitatum could be used as an alternatively practical genetic tool for conducting insertional mutagenesis in P. digitatum to study functional genomics.

  19. Characterization of rhizosphere bacteria for control of phytopathogenic fungi of tomato.

    Science.gov (United States)

    Pastor, Nicolás; Carlier, Evelin; Andrés, Javier; Rosas, Susana B; Rovera, Marisa

    2012-03-01

    Fluorescent Pseudomonas spp., isolated from rhizosphere soil of tomato and pepper plants, were evaluated in vitro as potential antagonists of fungal pathogens. Strains were characterized using the API 20NE biochemical system, and tested against the causal agents of stem canker and leaf blight (Alternaria alternata f. sp. lycopersici), southern blight (Sclerotium rolfsii Sacc.), and root rot (Fusarium solani). To this end, dual culture antagonism assays were carried out on 25% Tryptic Soy Agar, King B medium, and Potato Dextrose Agar to determine the effect of the strains on mycelial growth of the pathogens. The effect of two concentrations of FeCl(3) on antagonism against Alternaria alternata f. sp. lycopersici was also tested. In addition, strains were screened for ability to produce exoenzymes and siderophores. Finally, the selected Pseudomonas strain, PCI2, was evaluated for effect on tomato seedling development and as a potential candidate for controlling tomato damping-off caused by Sclerotium rolfsii Sacc., under growth chamber conditions. All strains significantly inhibited Alternaria alternata f. sp. lycopersici, particularly in 25% TSA medium. Antagonistic effect on Sclerotium rolfsii Sacc. and Fusarium solani was greater on King B medium. Protease was produced by 30% of the strains, but no strains produced cellulase or chitinase. Growth chamber studies resulted in significant increases in plant stand as well as in root dry weight. PCI2 was able to establish and survive in tomato plants rhizosphere after 40 days following planting of bacterized seeds. Copyright © 2011 Elsevier Ltd. All rights reserved.

  20. Resistances to an insect herbivore and a phytopathogen in Barbarea vulgaris

    DEFF Research Database (Denmark)

    Christensen, Stina

    The crucifer Barbarea vulgaris grows naturally in Europe and Western Asia. In Denmark and neighbouring countries, the subspecies arcuata exists in two types that differ in insect and pathogen resistance, as well as in several other traits. The types have been named after the degree of leaf...... that the distinctiveness of the two types is most likely maintained by a considerable reproductive barrier, and that selection against hybridisation occurs both before and after fertilisation....

  1. The F-box protein MAX2 contributes to resistance to bacterial phytopathogens in Arabidopsis thaliana.

    Science.gov (United States)

    Piisilä, Maria; Keceli, Mehmet A; Brader, Günter; Jakobson, Liina; Jõesaar, Indrek; Sipari, Nina; Kollist, Hannes; Palva, E Tapio; Kariola, Tarja

    2015-02-13

    The Arabidopsis thaliana F-box protein MORE AXILLARY GROWTH2 (MAX2) has previously been characterized for its role in plant development. MAX2 appears essential for the perception of the newly characterized phytohormone strigolactone, a negative regulator of polar auxin transport in Arabidopsis. A reverse genetic screen for F-box protein mutants altered in their stress responses identified MAX2 as a component of plant defense. Here we show that MAX2 contributes to plant resistance against pathogenic bacteria. Interestingly, max2 mutant plants showed increased susceptibility to the bacterial necrotroph Pectobacterium carotovorum as well as to the hemi-biotroph Pseudomonas syringae but not to the fungal necrotroph Botrytis cinerea. max2 mutant phenotype was associated with constitutively increased stomatal conductance and decreased tolerance to apoplastic ROS but also with alterations in hormonal balance. Our results suggest that MAX2 previously characterized for its role in regulation of polar auxin transport in Arabidopsis, and thus plant development also significantly influences plant disease resistance. We conclude that the increased susceptibility to P. syringae and P. carotovorum is due to increased stomatal conductance in max2 mutants promoting pathogen entry into the plant apoplast. Additional factors contributing to pathogen susceptibility in max2 plants include decreased tolerance to pathogen-triggered apoplastic ROS and alterations in hormonal signaling.

  2. Interactions between a cotton phytopathogen and the host using a genomics analysis

    Science.gov (United States)

    Cotton (Gossypium hirsutum L.) is an economic crop grown worldwide. Numerous G. hirsutum polyploids have been sequenced. Bacterial infections of cotton can cause major yield losses. Pantoea ananatis is a known bacterial pathogen of both cotton buds and bolls. Thus, we conducted a whole genome an...

  3. Whole cell fatty acid analysis as a tool for classification of phytopathogenic pseudomonas bacteria

    NARCIS (Netherlands)

    Janse, J.D.

    1992-01-01

    In this thesis some members of the plant pathogenic bacterial genus Pseudomonas have been studied. Conventional morphological, biochemical, physiological and pathogenitcity tests as well as a 'finger-print' technique, viz. automated whole cell fatty acid analysis, were

  4. Rhizospheric fungi of Panax notoginseng: diversity and antagonism to host phytopathogens

    Directory of Open Access Journals (Sweden)

    Cui-Ping Miao

    2016-04-01

    Conclusion: Our results suggest that diverse fungi including potential pathogenic ones exist in the rhizosphere soil of 2-yr-old P. notoginseng and that antagonistic isolates may be useful for biological control of pathogens.

  5. Identification and Pathogenicity of Phytopathogenic Bacteria Associated with Soft Rot Disease of Girasole Tuber

    Directory of Open Access Journals (Sweden)

    Mamdoh Ewis ISMAIL

    2012-02-01

    Full Text Available During 2010-2011 growing seasons six bacterial isolates were separated from naturally infected girasole plants tubers (Helianthus tuberosus L. cv. �Balady�, showing soft rot, collected from experimental Farm of the Faculty of Agriculture, in El-Minia University, Egypt. Pathogenicity tests showed various virulence for the bacteria isolated from girasole tubers, found pathogenic. These organisms were characterized as rod-shaped, Gram negative, ?-methyl-d-glucoside medium, reducing substances from sucrose, phos, phatase activity and deep cavities on pectate medium. Otherwise, diagnostic tests suggested that the pathogen was Erwinia carotovora ssp. carotovora. The isolated bacteria caused soft rot of wounded tubers when inoculated into tissues. The bacterial isolates were compared for their degree of pathogenicity as well as for differences in specific symptoms, induced in different hosts. The tested isolates could infect several host ranges, such as fruits of apricot, apple, olive, lemon, squash, eggplant and potato tubers, bulbs and garlic and onion cloves, roots radish, carrot, sweet potato and rape. On the other hand, no symptoms were exhibited on pods of bean and cowpea, faba bean, fruits of pepper and tomato. The extracts of experimentally diseased girasole tubers were active in pectinase and also in caboxymethyl cellulose at pH 6 compared to enzyme activities in healthy tissues. Also, the isolated bacteria increased the total and reducing sugars in infected tissues.

  6. Erwinia tasmaniensis sp. nov., a non-phytopathogenic bacterium from apple and pear trees.

    Science.gov (United States)

    Geider, Klaus; Auling, Georg; Du, Zhiqiang; Jakovljevic, Vladimir; Jock, Susanne; Völksch, Beate

    2006-12-01

    Bacteria were isolated from flowers and bark of apple and pear trees at three places in Australia. In Victoria, Tasmania and Queensland, strains with white colonies on nutrient agar were screened for dome-shaped colony morphology on agar with sucrose and were found to be closely related by several criteria. The isolates were not pathogenic on apples or pears. They were characterized by a polyphasic approach including microbiological and API assays as well as fatty acid methyl ester analysis, DNA-DNA hybridization and DNA sequencing. For molecular classification, the 16S rRNA cistron and the conserved genes gpd and recA of these bacteria were investigated. Together with other taxonomic criteria, the results of these studies indicate that the bacteria belong to a novel separate species, which we propose to name Erwinia tasmaniensis sp. nov., with the type strain Et1/99(T) (=DSM 17950(T)=NCPPB 4357(T)). From DNA-DNA hybridization kinetics, microbiological characteristics and nucleotide sequence analyses, this species is related to pathogenic Erwinia species, but also to the epiphytic species Erwinia billingiae.

  7. The Influence of Learning on Host Plant Preference in a Significant Phytopathogen Vector, Diaphorina citri.

    Directory of Open Access Journals (Sweden)

    Dara G Stockton

    Full Text Available Although specialist herbivorous insects are guided by innate responses to host plant cues, host plant preference may be influenced by experience and is not dictated by instinct alone. The effect of learning on host plant preference was examined in the Asian citrus psyllid, Diaphorina citri; vector of the causal agent of citrus greening disease or huanglongbing. We investigated: a whether development on specific host plant species influenced host plant preference in mature D. citri; and b the extent of associative learning in D. citri in the form of simple and compound conditioning. Learning was measured by cue selection in a 2-choice behavioral assay and compared to naïve controls. Our results showed that learned responses in D. citri are complex and diverse. The developmental host plant species influenced adult host plant preference, with female psyllids preferring the species on which they were reared. However, such preferences were subject to change with the introduction of an alternative host plant within 24-48 hrs, indicating a large degree of experience-dependent response plasticity. Additionally, learning occurred for multiple sensory modalities where novel olfactory and visual environmental cues were associated with the host plant. However, males and females displayed differing discriminatory abilities. In compound conditioning tasks, males exhibited recognition of a compound stimulus alone while females were capable of learning the individual components. These findings suggest D. citri are dynamic animals that demonstrate host plant preference based on developmental and adult experience and can learn to recognize olfactory and visual host plant stimuli in ways that may be sex specific. These experience-based associations are likely used by adults to locate and select suitable host plants for feeding and reproduction and may suggest the need for more tailored lures and traps, which reflect region-specific cultivars or predominate Rutaceae in the area being monitored.

  8. Blue mold to genomics and beyond: Insights into the biology and virulence of phytopathogenic Penicillium species

    Science.gov (United States)

    Pomes, mainly apples and pears, are economically important fruits produced and consumed worldwide. The United States is the second largest producer of pome fruit in the world behind China. Penicillium expansum and other Penicillium spp. are the most common fungal plant pathogens that cause blue mold...

  9. The ubiquitin conjugating enzyme, TaU4 regulates wheat defence against the phytopathogen Zymoseptoria tritici

    Science.gov (United States)

    Millyard, Linda; Lee, Jack; Zhang, Cunjin; Yates, Gary; Sadanandom, Ari

    2016-01-01

    Mycosphaerella graminicola (Zymoseptoria tritici commonly known as Septoria), the causal agent of Septoria Leaf Blotch (STB), is considered one of the major threats to European wheat production. Previous studies have shown the importance of ubiquitination in plant defence against a multitude of pathogens. However the ubiquitination machinery in wheat is under studied, particularly E2 enzymes that have the ability to control the ubiquitination and thereby the fate of many different target proteins. In this study we identify an E2 enzyme, Triticum aestivum Ubiquitin conjugating enzyme 4 (TaU4) that functions in wheat defence against Septoria. We demonstrate TaU4 to be a bona fide E2 enzyme through an E2 charging assay. TaU4 localises in both the cytoplasm and nucleus, therefore potentially interacting with E3 ligases and substrate proteins in multiple compartments. Virus Induced Gene Silencing of TaU4 in wheat leaves resulted in delayed development of disease symptoms, reduced Septoria growth and reproduction. We conclude that TaU4 is a novel negative regulator of defence against Septoria. PMID:27759089

  10. Draft Genome Sequence of Phytopathogenic Fungus Fusarium fujikuroi CF-295141, Isolated from Pinus sylvestris

    Science.gov (United States)

    Bertoni-Mann, Michele; Sánchez-Hidalgo, Marina; González-Menéndez, Victor

    2016-01-01

    Here, we report the draft genome sequence of a new strain of Fusarium fujikuroi, isolated from Pinus sylvestris, which was also found to produce the mycotoxin beauvericin. The Illumina-based sequence analysis revealed an approximate genome size of 44.2 Mbp, containing 164 secondary metabolite biosynthetic clusters. PMID:27795279

  11. Synthesis, Biological Evaluation, and Molecular Modeling Studies of New Oxadiazole-Stilbene Hybrids against Phytopathogenic Fungi

    Science.gov (United States)

    Jian, Weilin; He, Daohang; Song, Shaoyun

    2016-08-01

    Natural stilbenes (especially resveratrol) play important roles in plant protection by acting as both constitutive and inducible defenses. However, their exogenous applications on crops as fungicidal agents are challenged by their oxidative degradation and limited availability. In this study, a new class of resveratrol-inspired oxadiazole-stilbene hybrids was synthesized via Wittig-Horner reaction. Bioassay results indicated that some of the compounds exhibited potent fungicidal activity against Botrytis cinerea in vitro. Among these stilbene hybrids, compounds 11 showed promising inhibitory activity with the EC50 value of 144.6 μg/mL, which was superior to that of resveratrol (315.6 μg/mL). Remarkably, the considerably abnormal mycelial morphology was observed in the presence of compound 11. The inhibitory profile was further proposed by homology modeling and molecular docking studies, which showed the possible interaction of resveratrol and oxadiazole-stilbene hybrids with the cytochrome P450-dependent sterol 14α-demethylase from B. cinerea (BcCYP51) for the first time. Taken together, these results would provide new insights into the fungicidal mechanism of stilbenes, as well as an important clue for biology-oriented synthesis of stilbene hybrids with improved bioactivity against plant pathogenic fungi in crop protection.

  12. Effect of three nematicides on the growth of some phytopathogenic bacteria and fungi.

    Science.gov (United States)

    El-Khadem, M; Mehiar, F; Embabi, M S

    1977-01-01

    The effect of three nematicides, aldicarb, fensulfothion, and phenamiphos at four concentrations (1, 5, 25, and 125 ppm) was tested on the growth of five bacteria, Agrobacterium tumefaciens, Corynebacterium fascians, Erwinia carotovora, Pseudomonas solanacearum, and Streptomyces scabies and four fungi, Fusarium oxysporum f. sp. vasinfectum, Fusarium solani, Rhizoctonia solani, and Sclerotium bataticola. Of the bacteria, P. solanacearum was most affected by the chemicals at all concentrations, while E. carotovora was least affected. Fensulfothion was generally the most effective nematicide on the bacteria tested, while phenamiphos was the least effective. Similarly, the effect of the chemicals on the fungi tested varied greatly. F. solani and R. solani were generally most affected, followed by F. oxysporum, while S. bataticola was least affected. Of the chemicals tested, phenamiphos was generally the most effective, followed by fensulfothion, while aldicarb was the least effective.

  13. Advances in Cotton Breeding for Resistance to Fusarium and Verticillium Wilt in the Last Fifty Years in China

    Institute of Scientific and Technical Information of China (English)

    JIAN Gui-liang; MA Cun; ZHENG Chuan-lin; ZOU Ya-fei

    2003-01-01

    This review summarizes the main advances in cotton breeding for resistance to fusarium andverticillium wilt in the last fifty years in China. The topics discussed include main achievements, experiences,problems and countermeasures for solving.

  14. Functional Analysis of Autophagy Genes via Agrobacterium-Mediated Transformation in the Vascular Wilt Fungus Verticillium dahliae

    Institute of Scientific and Technical Information of China (English)

    Lei Zhou; Jun Zhao; Wangzhen Guo; Tianzhen Zhang

    2013-01-01

    Autophagy is a widely conserved intracellular process for degradation and recycling of proteins,organelles and cytoplasm in eukaryotic organisms and is now emerging as an important process in foliar infection by many plant pathogenic fungi.However,the role of autophagy in soil-borne fungal physiology and infection biology is poorly understood.Here,we report the establishment of an Agrobacterium tumefaciens-mediated transformation (ATMT) system and its application to investigate two autophagy genes,VdATG8 and VdATG12,by means of targeted gene replacement and complementadon.Transformation of a cotton-infecting Verticilliun dahliae strain Vd8 with a novel binary vector pCOM led to the production of 384 geneticin-resistant transformants per 1 × 106 conidia.V.dahliae mutants lacking either VdATG8 or VdATG12 exhibited reduced conidiation and impaired aerial hyphae production.Disease development on Arabidopsis plants was slightly delayed when inoculated with VdATG8 or VdATG12 gene deletion mutants,compared with the wildtype and gene complemented strains.Surprisingly,in vitro inoculation with unimpaired roots revealed that the abilities of root invasion were not affected in gene deletion mutants.These results indicate that autophagy is necessary for aerial hyphae development and plant colonization but not for root infection in V.dahliae.

  15. Genotypic Resistance of F1 Cotton Hybrids by Inoculation with Different Virulent Isolates of the Fungus Verticillium Dahliae Klebahn

    Institute of Scientific and Technical Information of China (English)

    AMANTURDIEV Alisher; KIM Robert

    2008-01-01

    @@ The plant pathogen Verticilliurn dahliae causes severe cotton losses in Uzbekistan.To create cotton varieties that are resistant to the more virulent races of V.dahliae we wanted to determine genotypic resistance of varieties and lines of cotton to more virulent isolates of different geographic populations of the fungus V.dahliae isolated from different cotton varieties in Uzbekistan and determine the inheritance of wilt resistance with hybrids of the first generation to various isolates of V.dahliae.

  16. Potential for using Verticillium albo-atrum as a biocontrol agent for tree-of-heaven (Ailanthus altissima)

    Science.gov (United States)

    Donald D. Davis; Matthew Kasson; Mark. Schall

    2011-01-01

    Extensive, unprecedented wilt and mortality of the highly invasive, exotic tree-of-heaven (Ailanthus altissima) occurred recently within mixed hardwood forests in south-central Pennsylvania. Until this study, the cause of the epidemic was unknown.

  17. Root hairs play a key role in the endophytic colonization of olive roots by Pseudomonas spp. with biocontrol activity.

    Science.gov (United States)

    Prieto, Pilar; Schilirò, Elisabetta; Maldonado-González, María Mercedes; Valderrama, Raquel; Barroso-Albarracín, Juan Bautista; Mercado-Blanco, Jesús

    2011-08-01

    The use of indigenous bacterial root endophytes with biocontrol activity against soil-borne phytopathogens is an environmentally-friendly and ecologically-efficient action within an integrated disease management framework. The earliest steps of olive root colonization by Pseudomonas fluorescens PICF7 and Pseudomonas putida PICP2, effective biocontrol agents (BCAs) against Verticillium wilt of olive (Olea europaea L.) caused by the fungus Verticillium dahliae Kleb., are here described. A gnotobiotic study system using in vitro propagated olive plants, differential fluorescent-protein tagging of bacteria, and confocal laser scanning microscopy analysis have been successfully used to examine olive roots-Pseudomonas spp. interactions at the single-cell level. In vivo simultaneous visualization of PICF7 and PICP2 cells on/in root tissues enabled to discard competition between the two bacterial strains during root colonization. Results demonstrated that both BCAs are able to endophytically colonized olive root tissues. Moreover, results suggest a pivotal role of root hairs in root colonization by both biocontrol Pseudomonas spp. However, colonization of root hairs appeared to be a highly specific event, and only a very low number of root hairs were effectively colonized by introduced bacteria. Strains PICF7 and PICP2 can simultaneously colonize the same root hair, demonstrating that early colonization of a given root hair by one strain did not hinder subsequent attachment and penetration by the other. Since many environmental factors can affect the number, anatomy, development, and physiology of root hairs, colonization competence and biocontrol effectiveness of BCAs may be greatly influenced by root hair's fitness. Finally, the in vitro study system here reported has shown to be a suitable tool to investigate colonization processes of woody plant roots by microorganisms with biocontrol potential.

  18. Composition and antifungal activity of essential oils of Mentha piperita and Lavendula angustifolia on post-harvest phytopathogens.

    Science.gov (United States)

    Behnam, S; Farzaneh, M; Ahmadzadeh, M; Tehrani, A Sharifi

    2006-01-01

    The general antifungal activity of essential oils is well documented. The advantage of essential oils is their bioactivity in the vapor phase, a characteristic that makes them attractive as possible fumigants for stored product protection. Essential oils of aerial parts of Mentha piperita and Lavendula angustifolia were obtained with hydrodistillation and oils composition identified with GC-MS. Menthanol (36.24%) and menthone (32.42%) were the major compounds of the M. piperata essential oil. The essential oil of L. angustifolia was rich in linalool (49.2%) , linalyl acetate (12.3%), Lavendul acetate (6.5%), 4-terpineol (5.9%). Fungal toxicity of the essential oils were evaluated against three pathogenic fungi (Rhizopus stolonifer, Botrytis cinerea and Aspergillus niger) in vitro. Plate assayes showed that the different concentrations of essential oils have antifungal activity against these fungi, and the essential oil of L. angustifolia showed stronger fungistatic activity. Lavendula oil exhibited complete growth inhibition of all pathogens at 1000 ppm and minimum EC50 (311.24 ppm) resulted on B. cinerea.

  19. Kunitz Trypsin Inhibitor: An Antagonist of Cell Death Triggered by Phytopathogens and Fumonisin B1 in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Jing Li; Günter Brader; E. Tapio Palva

    2008-01-01

    Programmed cell death (PCD) is a central regulatory process in both plant development and in plant responses to pathogens. PCD requires a coordinate activation of pro-apoptotic factors such as proteases and suppressors inhibiting and modulating these processes. In plants, various caspase-like cysteine proteases as well as serine proteases have been implicated in PCD. Here, we show that a serine protease (Kunitz trypsin) inhibitor (KTI1) of Arabidopsis acts as a functional KTI when produced in bacteria and in planta. Expression of AtKTI1 is induced late in response to bacterial and fungal elicitors and to salicylic acid. RNAi silencing of the AtKTI1 gene results in enhanced lesion development after infiltration of leaf tissue with the PCD-eliciting fungal toxin fumonisin B1 (FB1) or the avirulent bacterial pathogen Pseudomonas syringae pv tomato DC3000 carrying avrB (Pst avrB). Overexpression of AtKTI1 results in reduced lesion development after Pst avrB and FB1 infiltration. Interestingly, RNAi silencing of AtKTI1 leads to enhanced resistance to the virulent pathogen Erwinia carotovora subsp, carotovora SCC1, while overexpression of AtKTI1 leads to higher susceptibility towards this pathogen. Together, these data indicate that AtKTI1 is involved in modulating PCD in plant-pathogen interactions.

  20. Microbiological Control of Soil-Borne Phytopathogenic Fungi with Special Emphasis on Wilt-Inducing Fusarium oxysporum

    National Research Council Canada - National Science Library

    Claude Alabouvette; Chantal Olivain; Quirico Migheli; Christian Steinberg

    2009-01-01

    .... In addition to Pseudomonas spp. and Trichoderma spp., which are the two most widely studied groups of biological control agents, the protective strains of Fusarium oxysporum represent an original model...

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

    Directory of Open Access Journals (Sweden)

    Raquel eGonzález-Fernández

    2015-10-01

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

  2. Improved production of phleichrome from the phytopathogenic fungus Cladosporium phlei using synthetic inducers and photodynamic ROS production by phleichrome.

    Science.gov (United States)

    So, Kum-Kang; Jo, Ik-Su; Chae, Min-Seon; Kim, Jung-Mi; Chung, Hea-Jong; Yang, Moon-Sik; Kim, Beom-Tae; Kim, Jin-Kug; Choi, Jong-Kyung; Kim, Dae-Hyuk

    2015-03-01

    Two different diketopiperazines, cyclo-(L-Pro-L-Leu) and cyclo-(L-Pro-L-Phe), which were isolated from the culture filtrate of Epichloe typhina and found to be inducers of phleichrome production, were chemically synthesized and evaluated for use in the improved production of phleichrome from wild-type and UV-mutagenized strains (M0035) of Cladosporium phlei. When supplemented with PDA and V8 juice agar media, both inducers showed significant increases in the production of phleichrome. Phleichrome production was increased in a dose-dependent manner up to a concentration of maximum yield for both inducers. No further significant induction was observed by supplementing inducers over the concentration of maximum yield. Among the two inducers, cyclo-(L-Pro-L-Phe) showed better inducing capability than cyclo-(L-Pro-L-Leu). The maximum yield was observed from the M0035 strain grown on V8 juice media supplemented with 150 μM cyclo-(L-Pro-L-Phe), which was estimated to be 232.6 mg of phleichrome per gram of mycelia and 10.2 mg of secreted phleichrome per 20 agar-plugs. Interestingly, growth inhibition was observed on V8 juice agar media with 100, 150, and 200 μM cyclo-(L-Pro-L-Phe) but not on PDA with the same amount of inducer, which suggests that the inhibitory effect might be through the overproduction of phleichrome rather than the toxic effect of the inducer itself. Superoxide production by purified phleichrome was dramatically stimulated upon illumination, thus demonstrating photodynamic production of superoxide in vitro by phleichrome.

  3. Rapid screening of an ordered fosmid library to clone multiple polyketide synthase genes of the phytopathogenic fungus Cladosporium phlei.

    Science.gov (United States)

    So, Kum-Kang; Kim, Jung-Mi; Nguyen, Ngoc-Luong; Park, Jin-Ah; Kim, Beom-Tae; Park, Seung-Moon; Hwang, Ki-Jun; Kim, Dae-Hyuk

    2012-12-01

    In previous studies, the biological characteristics of the fungus Cladosporium phlei and its genetic manipulation by transformation were assessed to improve production of the fungal pigment, phleichrome, which is a fungal perylenequinone that plays an important role in the production of a photodynamic therapeutic agent. However, the low production of this metabolite by the wild-type strain has limited its application. Thus, we attempted to clone and characterize the genes that encode polyketide synthases (PKS), which are responsible for the synthesis of fungal pigments such as perylenequinones including phleichrome, elsinochrome and cercosporin. Thus, we performed genomic DNA PCR using 11 different combinations of degenerate primers targeting conserved domains including β-ketoacyl synthase and acyltransferase domains. Sequence comparison of the PCR amplicons revealed a high homology to known PKSs, and four different PKS genes showing a high similarity to three representative types of PKS genes were amplified. To obtain full-length PKS genes, an ordered gene library of a phleichrome-producing C. phlei strain (ATCC 36193) was constructed in a fosmid vector and 4800 clones were analyzed using a simple pyramidal arrangement system. This hierarchical clustering method combines the efficiency of PCR with enhanced specificity. Among the three representative types of PKSs, two reducing, one partially reducing, and one non-reducing PKS were identified. These genes were subsequently cloned, sequenced, and characterized. Biological characterization of these genes to determine their roles in phleichrome production is underway, with the ultimate aim of engineering this pathway to overproduce the desired substance.

  4. [Biochemical basis of tolerance to osmotic stress in phytopathogenic fungus: The case of Macrophomina phaseolina (Tassi) Goid.

    Science.gov (United States)

    Martínez-Villarreal, Rodolfo; Garza-Romero, Tamar S; Moreno-Medina, Víctor R; Hernández-Delgado, Sanjuana; Mayek-Pérez, Netzahualcoyotl

    Fungus Macrophomina phaseolina (Tassi) Goid. is the causative agent of charcoal rot disease which causes significant yield losses in major crops such as maize, sorghum, soybean and common beans in Mexico. This fungus is a facultative parasite which shows broad ability to adapt itself to stressed environments where water deficits and/or high temperature stresses commonly occur. These environmental conditions are common for most cultivable lands throughout Mexico. Here we describe some basic facts related to the etiology and epidemiology of the fungus as well as to the importance of responses to stressed environments, particularly to water deficits, based on morphology and growth traits, as well as on physiology, biochemistry and pathogenicity of fungus M. phaseolina. To conclude, we show some perspectives related to future research into the genus, which emphasize the increasing need to improve the knowledge based on the application of both traditional and biotechnological tools in order to elucidate the mechanisms of resistance to environmental stress which can be extrapolated to other useful organisms to man. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  5. Clavibacter michiganensis subsp. michiganensis: first steps in the understanding of virulence of a Gram-positive phytopathogenic bacterium.

    Science.gov (United States)

    Gartemann, Karl-Heinz; Kirchner, Oliver; Engemann, Jutta; Gräfen, Ines; Eichenlaub, Rudolf; Burger, Annette

    2003-12-19

    Clavibacter michiganensis subsp. michiganensis is a plant-pathogenic actinomycete. It infects tomato, spreads through the xylem and causes bacterial wilt and canker. The wild-type strain NCPPB382 carries two plasmids, pCM1 and pCM2. The cured plasmid-free derivative CMM100 is still able to colonize tomato, but no disease symptoms develop indicating that all genes required for successful infection, establishment and growth in the plant reside on the chromosome. Both plasmids carry one virulence factor, a gene encoding a cellulase, CelA in case of pCM1 and a putative serine protease Pat-1 on pCM2. These genes can independently convert the non-virulent strain CMM100 into a pathogen causing wilt on tomatoes. Currently, genome projects for Cmm and the closely related potato-pathogen C. michiganensis subsp. sepedonicus have been initiated. The data from the genome project shall give clues on further genes involved in plant-microbe interaction that can be tested experimentally. Especially, identification of genes related to host-specificity through genome comparison of the two subspecies might be possible.

  6. Stable transformation of the gram-positive phytopathogenic bacterium Clavibacter michiganensis subsp. sepedonicus with several cloning vectors.

    Science.gov (United States)

    Laine, M J; Nakhei, H; Dreier, J; Lehtilä, K; Meletzus, D; Eichenlaub, R; Metzler, M C

    1996-05-01

    In this paper we describe transformation of Clavibacter michiganensis subsp. sepedonicus, the potato ring rot bacterium, with plasmid vectors. Three of the plasmids used, pDM100, pDM302, and pDM306, contain the origin of replication from pCM1, a native plasmid of C. michiganensis subsp. michiganensis. We constructed two new cloning vectors, pHN205 and pHN216, by using the origin of replication of pCM2, another native plasmid of C. michiganensis subsp. michiganensis. Plasmids pDM302, pHN205, and pHN216 were stably maintained without antibiotic selection in various strains of C. michiganensis subsp. sepedonicus. We observed that for a single plasmid, different strains of C. michiganensis subsp. sepedonicus showed significantly different transformation efficiencies. We also found unexplained strain-to-strain differences in stability with various plasmid constructions containing different arrangements of antibiotic resistance genes and origins of replication. We examined the effect of a number of factors on transformation efficiency. The best transformation efficiencies were obtained when C. michiganensis subsp. sepedonicus cells were grown on DM agar plates, harvested during the early exponential growth phase, and used fresh (without freezing) for electroporation. The maximal transformation efficiency obtained was 4.6 x 10(4) CFU/microgram of pHN216 plasmid DNA. To demonstrate the utility of this transformation system, we cloned a beta-1,4-endoglucanase-encoding gene from C. michiganensis subsp. sepedonicus into pHN216. When this construction, pHN216:C8, was electroporated into competent cells of a cellulase-deficient mutant, it restored cellulase production to almost wild-type levels.

  7. Evidence for plasmid-encoded virulence factors in the phytopathogenic bacterium Clavibacter michiganensis subsp. michiganensis NCPPB382.

    Science.gov (United States)

    Meletzus, D; Bermphol, A; Dreier, J; Eichenlaub, R

    1993-01-01

    The tomato pathogen Clavibacter michiganensis subsp. michiganensis NCPPB382, which causes bacterial wilt, harbors two plasmids pCM1 (27.5 kb) and pCM2 (72 kb). After curing of the plasmids, bacterial derivatives were still proficient in the ability to colonize the host plant and in the production of exopolysaccharides but exhibited a reduced virulence. When one of the two plasmids is lost, there is a significant delay in the development of wilting symptoms after infection and a plasmid-free derivative is not able to induce disease symptoms. By cloning of restriction fragments of both plasmids in the plasmid-free strain CMM100, two DNA fragments which restored the virulent phenotype were identified. Further analysis suggested that a fragment of plasmid pCM1 encodes an endocellulase which is involved in the expression of the pathogenic phenotype. Images PMID:8458855

  8. Large-scale gene discovery in the oomycete Phytophthora infestans reveals likely components of phytopathogenicity shared with true fungi

    NARCIS (Netherlands)

    Randall, T.A.; Dwyer, R.A.; Huitema, E.; Beyer, K.; Cvitanich, C.; Kelkar, H.; Ah Fong, A.M.V.; Gates, K.; Roberts, S.; Yatzkan, E.; Gaffney, T.; Law, M.; Testa, A.; Torto-Alalibo, T.; Zhang Meng,; Zheng Li,; Mueller, E.; Windass, J.; Binder, A.; Birch, P.R.J.; Gisi, U.; Govers, F.; Gow, N.A.; Mauch, F.; West, van P.; Waugh, M.E.; Yu Jun,; Boller, T.; Kamoun, S.; Lam, S.T.; Judelson, H.S.

    2005-01-01

    o overview the gene content of the important pathogen Phytophthora infestans, large-scale cDNA and genomic sequencing was performed. A set of 75,757 high-quality expressed sequence tags (ESTs) from P. infestans was obtained from 20 cDNA libraries representing a broad range of growth conditions, stre

  9. Comparison of the effects of chlorite-oxidized oxyamylose and polyacrylic acid on the multiplication of phytopathogenic viruses.

    Science.gov (United States)

    Kluge, S

    1985-10-01

    Polyacrylic acid (PAA) and chlorite-oxidized oxyamylose (COAM) inhibit the multiplication of tobacco mosaic virus (TMV) in leaf disks by up to 50%. The reduction in TMV content is time-dependent and decreases with longer time intervals between the virus infection and the application of substances. The multiplication of potato virus X (PVX) in leaf disks is not affected by either PAA or COAM. In intact plants PAA produces a strong antiviral effect on both PVX and red clover mottle virus (RCMV). The effect produced by COAM is much less pronounced, although this substance is less toxic and could be used in a higher concentration than PAA. Neither of these compounds has a significant influence on the development of virus-induced necroses in Nicotiana glutinosa, Gomphrena globosa or Phaseolus vulgaris plants when administered one day before or after virus infection.

  10. Nucleotide Sequence and Evolution of the Five-Plasmid Complement of the Phytopathogen Pseudomonas syringae pv. maculicola ES4326

    OpenAIRE

    2004-01-01

    Plasmids are transmissible, extrachromosomal genetic elements that are often responsible for environmental or host-specific adaptations. In order to identify the forces driving the evolution of these important molecules, we determined the complete nucleotide sequence of the five-plasmid complement of the radish and Arabidopsis pathogen Pseudomonas syringae pv. maculicola ES4326 and conducted an intraspecific comparative genomic analysis. To date, this is the most complex fully sequenced plasm...

  11. Erwinia amylovora novel plasmid pEI70: complete sequence, biogeography, and role in aggressiveness in the fire blight phytopathogen.

    Directory of Open Access Journals (Sweden)

    Pablo Llop

    Full Text Available Comparative genomics of several strains of Erwinia amylovora, a plant pathogenic bacterium causal agent of fire blight disease, revealed that its diversity is primarily attributable to the flexible genome comprised of plasmids. We recently identified and sequenced in full a novel 65.8 kb plasmid, called pEI70. Annotation revealed a lack of known virulence-related genes, but found evidence for a unique integrative conjugative element related to that of other plant and human pathogens. Comparative analyses using BLASTN showed that pEI70 is almost entirely included in plasmid pEB102 from E. billingiae, an epiphytic Erwinia of pome fruits, with sequence identities superior to 98%. A duplex PCR assay was developed to survey the prevalence of plasmid pEI70 and also that of pEA29, which had previously been described in several E. amylovora strains. Plasmid pEI70 was found widely dispersed across Europe with frequencies of 5-92%, but it was absent in E. amylovora analyzed populations from outside of Europe. Restriction analysis and hybridization demonstrated that this plasmid was identical in at least 13 strains. Curing E. amylovora strains of pEI70 reduced their aggressiveness on pear, and introducing pEI70 into low-aggressiveness strains lacking this plasmid increased symptoms development in this host. Discovery of this novel plasmid offers new insights into the biogeography, evolution and virulence determinants in E. amylovora.

  12. Erwinia amylovora novel plasmid pEI70: complete sequence, biogeography, and role in aggressiveness in the fire blight phytopathogen.

    Science.gov (United States)

    Llop, Pablo; Cabrefiga, Jordi; Smits, Theo H M; Dreo, Tanja; Barbé, Silvia; Pulawska, Joanna; Bultreys, Alain; Blom, Jochen; Duffy, Brion; Montesinos, Emilio; López, María M

    2011-01-01

    Comparative genomics of several strains of Erwinia amylovora, a plant pathogenic bacterium causal agent of fire blight disease, revealed that its diversity is primarily attributable to the flexible genome comprised of plasmids. We recently identified and sequenced in full a novel 65.8 kb plasmid, called pEI70. Annotation revealed a lack of known virulence-related genes, but found evidence for a unique integrative conjugative element related to that of other plant and human pathogens. Comparative analyses using BLASTN showed that pEI70 is almost entirely included in plasmid pEB102 from E. billingiae, an epiphytic Erwinia of pome fruits, with sequence identities superior to 98%. A duplex PCR assay was developed to survey the prevalence of plasmid pEI70 and also that of pEA29, which had previously been described in several E. amylovora strains. Plasmid pEI70 was found widely dispersed across Europe with frequencies of 5-92%, but it was absent in E. amylovora analyzed populations from outside of Europe. Restriction analysis and hybridization demonstrated that this plasmid was identical in at least 13 strains. Curing E. amylovora strains of pEI70 reduced their aggressiveness on pear, and introducing pEI70 into low-aggressiveness strains lacking this plasmid increased symptoms development in this host. Discovery of this novel plasmid offers new insights into the biogeography, evolution and virulence determinants in E. amylovora.

  13. Reactive oxygen species and autophagy play a role in survival and differentiation of the phytopathogen Moniliophthora perniciosa.

    Science.gov (United States)

    Pungartnik, C; Melo, S C O; Basso, T S; Macena, W G; Cascardo, J C M; Brendel, M

    2009-01-01

    The hemibiotrophic basidiomycete Moniliophthora perniciosa causes "witches' broom disease" in cacao (Theobroma cacao). During plant infection, M. perniciosa changes from mono to dikaryotic life form, an event which could be triggered by changes in plant nutritional offer and plant defense molecules, i.e., from high to low content of glycerol and hydrogen peroxide. We have recently shown that in vitro glycerol induces oxidative stress resistance in dikaryotic M. perniciosa. In order to understand under which conditions in parasite-plant interaction M. perniciosa changes from intercellular monokaryotic to intracellular dikaryotic growth phase we studied the role of glycerol on mutagen-induced oxidative stress resistance of basidiospores and monokaryotic hyphae; we also studied the role of H(2)O(2) as a signaling molecule for in vitro dikaryotization and whether changes in nutritional offer by the plant could be compensated by inducible fungal autophagy. Mono-/dikaryotic glycerol or glucose-grown cells and basidiospores were exposed to the oxidative stress-inducing mutagens H(2)O(2) and Paraquat as well as to pre-dominantly DNA damaging 4-nitroquinoline-1-oxide and UVC irradiation. Basidiospores showed highest resistance to all treatments and glycerol-grown monokaryotic hyphae were more resistant than dikaryotic hyphae. Monokaryotic cells exposed to 1microM of H(2)O(2) in glycerol-media induced formation of clamp connections within 2 days while 1mM H(2)O(2) did not within a week in the same medium; no clamp connections were formed in H(2)O(2)-containing glucose media within a week. Lower concentrations of H(2)O(2) and glycerol, when occurring in parallel, are shown to be two signals for dikaryotization in vitro and may be also during the course of infection. Q-PCR studies of glycerol-grown dikaryotic cells exposed to oxidative stress (10mM H(2)O(2)) showed high expression of MpSOD2 and transient induction of ABC cytoplasmic membrane transporter gene MpYOR1 and autophagy-related gene MpATG8. Expression of a second ABC transporter gene MpSNQ2 was 14-fold induced after H(2)O(2) exposure in glucose as compared to glycerol-grown hyphae while MpYOR1 did not show strong variation of expression under similar conditions. Glucose-grown dikaryotic cells showed elevated expression of MpATG8, especially after exposure to H(2)O(2) and 4-nitroquinoline-1-oxide. During different stages preceding basidiocarp formation MpATG8 and the two catalase-encoding genes MpCTA1 and MpCTT1 were expressed continuously. We have compiled our results and literature data in a model graph, which compares the in vitro and in planta development and differentiation of M. perniciosa with the help of physiological and morphological landmarks.

  14. An In Vitro Attempt for Controlling Severe Phytopathogens and Human Pathogens Using Essential Oils from Mediterranean Plants of Genus Schinus.

    Science.gov (United States)

    Elshafie, Hazem Salaheldin; Ghanney, Nadia; Mang, Stefania Mirela; Ferchichi, Ali; Camele, Ippolito

    2016-03-01

    Growing concerns about food safety and environmental protection enhanced the need for new and safe plant disease control strategies. The chemical composition of the three essential oils (EOs) extracted from leaves and fruits of Schinus terebinthifolius and leaves of Schinus molle, growing in Tunisia, was studied by GC and GC-MS. In all, 12 compounds were identified. The oils were mainly composed of terpene compounds. α-Pinene, α-phellandrene, and D-limonene were the major constituents. The aim of the current study was to evaluate the in vitro antimicrobial effectiveness of three EOs derived from plants of genus Schinus and extracted from leaves and fruits of S. terebinthifolius and leaves of S. molle. Both antifungal and antibacterial activities of the EOs were examined. The antifungal activity of the studied EOs was investigated against Colletotrichum acutatum and Botrytis cinerea in comparison with the systemic fungicide azoxystrobin used at 0.8 μL mL(-1). The antibacterial activity was evaluated against three strains of Gram-positive (G+ve) bacteria (Bacillus megaterium, Bacillus mojavensis and Clavibacter michiganensis) and four strains of Gram-negative (G-ve) bacteria (Escherichia coli, Xanthomonas campestris, Pseudomonas savastanoi, and Pseudomonas syringae pv. phaseolicola) compared with the synthetic antibiotic tetracycline at a concentration of 1600 μg mL(-1). The minimum inhibitory concentration of the studied EOs has been evaluated against the above microorganisms using the 96-well microplate method. Tested microorganisms exhibited different levels of sensitivity to each tested EO. All investigated EOs reduced the fungal mycelial growth when used at low concentrations from 250 to 1000 ppm and from 2000 to 8000 ppm against C. acutatum and B. cinerea, respectively. Higher concentrations of the same EOs exhibited a fungicidal effect against both mitosporic fungi. The EO extracted from leaves of S. terebinthifolius significantly inhibited the growth of tested bacterial strains. Nevertheless, E. coli showed a weak resistance toward the same EO and a high resistance toward the other two tested EOs. Finally, P. savastanoi and P. syringae pv. phaseolicola showed a high resistance toward all tested EOs.

  15. Horizontal gene transfer confers adaptive advantages to phytopathogenic fungi: a case study of catalase-peroxidase in Fusarium verticillioides

    Science.gov (United States)

    Horizontal gene transfer (HGT), the exchange and stable integration of genetic material between different evolutionary lineages, is widely observed in fungi. We hypothesize that successful stabilization of HGT elements provides adaptive advantages (e.g., virulence). Catalase/peroxidases (KatGs) are ...

  16. Antifungal efficiency of a lipopeptide biosurfactant derived from Bacillus subtilis SPB1 versus the phytopathogenic fungus, Fusarium solani.

    Science.gov (United States)

    Mnif, Ines; Hammami, Ines; Triki, Mohamed Ali; Azabou, Manel Cheffi; Ellouze-Chaabouni, Semia; Ghribi, Dhouha

    2015-11-01

    Bacillus subtilis SPB1 lipopeptides were evaluated as a natural antifungal agent against Fusarium solani infestation. In vitro antifungal assay showed a minimal inhibitory concentration of about 3 mg/ml with a fungicidal mode of action. In fact, treatment of F. solani by SPB1 lipopeptides generated excessive lyses of the mycelium and caused polynucleation and destruction of the related spores together with a total inhibition of spore production. Furthermore, an inhibition of germination potency accompanied with a high spore blowing was observed. Moreover, in order to be applied in agricultural field, in vivo antifungal activity was proved against the dry rot potato tubers caused by F. solani. Preventive treatment appeared as the most promising as after 20 days of fungi inoculation, rot invasion was reduced by almost 78%, in comparison to that of non-treated one. When treating infected tomato plants, disease symptoms were reduced by almost 100% when applying the curative method. Results of this study are very promising as it enables the use of the crude lipopeptide preparation of B. subtilis SPB1 as a potent natural fungicide that could effectively control the infection of F. solani in tomato and potato tubers at a concentration similar to the commercial fungicide hymexazol and therefore prevent the damage of olive tree.

  17. [Study of the effect of volatile metabolites of Trichoderma hamatum on the growth of phytopathogenic soilborne fungi.].

    Science.gov (United States)

    Dal Bello, G M; Mónaco, C I; Cháves, A R

    1997-09-01

    Volatile compounds produced by Trichoderma hamatum were tested for their capacity to suppress in vitro the growth of Alternaria citri, Bipolaris cynodontis, Bipolaris sorokiniana, Curvularia brachyspora, Curvularia lunata, Curvularia oryzae-sativae, Drechslera tritici-repentis, Rhizoctonia solani, Sclerotinia minor and Sclerotium rolfsii. The organisms were cultured in an apparatus made with two Erlenmeyer flasks assembled by their top parts. With the aid of the gas chromatographic technique the variation of carbon dioxide, oxygen and ethylene in the internal system was determined. Acetaldehyde and ethanol were not found. Due to the respiratory metabolism of T. hamatum the carbon dioxide level progressively increased while the oxygen content decreased. Ethylene production was low and after three days remained constant. Excepting C. oryzae-sativae and B. cynodontis the other species showed changes in the growth and development. These results suggest the inhibitory volatiles of T. hamatum as one possible mechanism of biological control.

  18. Development of alternative strategies for the control of the important phytopathogens Phytophthora infestans (Mont.) and Erwinia amylovora (Burrill)

    OpenAIRE

    2007-01-01

    Within the framework of two projects, financed by DBU (Deutsche Bundesstiftung Umwelt) and ProInno ("Förderung der Erhöhung der Innovationskompetenz mittelständischer Unternehmen"), respectively, in co-operation with an industrial partner, alternative phytosanitary compounds from natural sources have been screened. High throughput screening systems were developed and used for testing of large numbers of extracts of Actinomycetes in 96-well multiplates against Phytophthora infestans and Erwini...

  19. Production of antioomycete compounds active against the phytopathogens Phytophthora sojae and Aphanomyces cochlioides by clavicipitoid entomopathogenic fungi.

    Science.gov (United States)

    Putri, Sastia Prama; Ishido, Kei-Ichi; Kinoshita, Hiroshi; Kitani, Shigeru; Ihara, Fumio; Sakihama, Yasuko; Igarashi, Yasuhiro; Nihira, Takuya

    2014-05-01

    A total of 412 strains belonging to 14 genera of clavicipitoid entomopathogenic fungi (EPF) were screened for activities against two economically important plant pathogenic oomycetes, Phytophthora sojae and Aphanomyces cochlioides. To identify the antioomycete compounds produced by EPF, the extracts of 13 highly active EPF strains were characterized in detail by high performance liquid chromatography with diode array detection and high-resolution mass spectrometric detection and antioomycete assay. The antioomycete activity of several Metarhizium extracts was associated with previously isolated aurovertins, fungerin, N-(methyl-3-oxodec-6-enoyl)-2-pyrroline, and N-(methyl-3-oxodecanoyl)-2-pyrroline. The depsipeptide beauvericin was confirmed to be one of the active principles of three strains of Isaria tenuipes, which strongly inhibited mycelial growth of both P. sojae and A. cochlioides. Two known bioactive metabolites, paecilosetin and aranorosinol A, together with a novel and potent antioomycete compound, farinomalein, were isolated from the extracts of Isaria farinosa and all compounds were confirmed to have antioomycete activity. Identification of 8 antioomycete compounds from 13 clavicipitioid EPF demonstrated a new potential use of EPF as a source of compounds for the control of soil-borne plant pathogenic oomycetes.

  20. Isolation and sequence analysis of Clpg1, a gene coding for an endopolygalacturonase of the phytopathogenic fungus Colletotrichum lindemuthianum.

    Science.gov (United States)

    Centis, S; Dumas, B; Fournier, J; Marolda, M; Esquerré-Tugayé, M T

    1996-04-17

    Oligodeoxyribonucleotide primers designed from the N-terminal amino acid (aa) sequence of the endopolygalacturonase (EndoPG) of Colletotrichum lindemuthianum (Cl) race beta and from an internal sequence conserved among different fungal EndoPG were used in a polymerase chain reaction (PCR) to amplify genomic related sequences of the fungus. A 542-bp fragment, designated pgA, was obtained and used as a probe to screen a partial genomic library of Cl. Among the positive clones, one was further analyzed. Nucleotide sequencing of this clone revealed on ORF encoding a 363-amino-acid (aa) polypeptide beginning with a signal peptide of 26 aa interrupted by an intron of 70 bp, and showing a high degree of homology to ten fungal EndoPG sequences. Consensus sequences were identified in the 5' non-coding region. This genomic clone was thereafter designated Clpg1. Southern analysis, performed with a Clpg1-specific probe, showed that this gene is present as a single copy in the Cl genome.