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1.
Monoclonal and polyclonal antibodies that bind to eggs and/or second-stage juveniles of the nematode Meloidogyne javanica were tested for their effects on the parasitic interactions between this nematode and the fungus Trichoderma. Parasitism of Trichoderma asperellum-203 and Trichoderma atroviride on nematode egg masses, eggs and juveniles was enhanced when antibodies were incorporated into in vitro parasitism bioassays. Parasitism on separated eggs (without gelatinous matrix) and their hatched juveniles was also improved, compared to controls without antibodies that did not attach fungal conidia. Improved parasitism could be due to bilateral binding of the antibodies to the nematodes and conidia, enabling better conidial attachment to the nematodes. Enhanced germination of antibody-bound conidia further improved parasitism. Differences were observed among antibodies in their effects on fungal parasitism and their interaction with Trichoderma species. We focused mainly on the egg- and juvenile-binding monoclonal antibody MISC that exhibited a stronger reaction with T. asperellum-203 than with T. atroviride. Pretreatment of this antibody with fucose inhibited its binding to nematodes and conidial attachment to nematodes, as well as conidial agglutination in the presence of the antibody. Antibody binding to juveniles affected their movement and viability, especially gelatinous matrix-originated juveniles. The fucose-specific lectin Ulex europaeus-I enhanced conidial attachment to nematode life-stages, and conidial agglutination occurred in its presence. These phenomena were inhibited by preincubating lectin with fucose. Our results suggest that carbohydrate residues, such as fucose, on the surface of the nematode and fungal conidia are involved in the antibody- and lectin-mediated improved parasitism.  相似文献   

2.
Sixteen isolates belonging to 11 species of Trichoderma (T. asperellum, T. ceramicum, T. andinensis, T. orientalis, T. atroviride, T. viridescens, T. brevicompactum, T. harzianum, T. virens, T. koningii and T. koningiopsis) were evaluated for biological control of potato (Solanum tuberosum) stem rot caused by Sclerotinia sclerotiorum. In dual culture tests, all antagonists significantly reduced sclerotia formation, and were able to inhibit radial growth of the pathogen. Growth inhibition by production of volatile and non-volatile inhibitors was also measured in in vitro tests. In screening the most efficient species of Trichoderma, establishment of mycelium on sclerotia and sclerotia lysis were also considered as important biocontrol qualities. Excluding T. asperellum, T. brevicompactum, T. andinensis and T. harzianum, all tested Trichoderma species were able to lyse sclerotia. The sclerotia-destroying species of Trichoderma and one isolate of Talaromyces flavus were tested in greenhouse tests and during 2 years of field experimentation during the 2007 and 2008 cropping seasons. After one aerial application of spore suspension in greenhouse trials, T. koningii, T. virens, T. ceramicum and T. viridescens were the most effective bio-agents and reduced significantly disease severity, and the least biocontrol efficacy was observed in T. flavus. Under field conditions and after five soil and foliar applications of spore suspension, all tested antagonists reduced significantly disease incidence. T. viridescens followed by T. ceramicum showed the best results. T. flavus and T. orientalis were less effective than other tested antagonists in both field trials.  相似文献   

3.
Twenty-one isolates of Trichoderma spp. were collected from eight states in Colombia and characterized based on the 5′ end of the translation elongation factor-1α (EF1-α1) gene and RNA polymerase II gene encoding the second largest protein subunit (RPB2) by using mixed primers. Seven species of soil-dwelling Trichoderma were found: T. atroviride, T. koningiopsis, T. asperellum, T. spirale, T. harzianum, T. brevicompactum and T. longibrachiatum. Species identifications based on the EF1-α1 gene were consistent with those obtained from the RPB2 gene. Phylogenetic analyses with high bootstrap values supported the validity of the identification of all isolates. These results suggest that using the combination of the genes EF1-α1 and RPB2 is highly reliable for molecular characterization of Trichoderma species. Trichoderma asperellum Th034, T. atroviride Th002 and T. harzianum Th203 prevented germination of more than 70 % of sclerotia of Sclerotinia sclerotiorum in bioassay tests and are promising biological control agents. No relationship between mycelium growth rate and parasitism level was found.  相似文献   

4.
Trichoderma spp. are common soil fungi used as biocontrol agents due to their capacity to produce antibiotics, induce systemic resistance in plants and parasitize phytopathogenic fungi of major agricultural importance. The present study investigated whether colonization of Arabidopsis thaliana seedlings by Trichoderma atroviride affected plant growth and development. Here it is shown that T. atroviride promotes growth in Arabidopsis. Moreover, T. atroviride produced indole compounds in liquid cultures. These results suggest that indoleacetic acid-related indoles (IAA-related indoles) produced by T. atroviride may have a stimulatory effect on plant growth. In addition, whether colonization of Arabidopsis roots by T. atroviride can induce systemic protection against foliar pathogens was tested. Arabidopsis roots inoculation with T. atroviride provided systemic protection to the leaves inoculated with bacterial and fungal pathogens. To investigate the possible pathway involved in the systemic resistance induced by T. atroviride, the expression profile of salicylic acid, jasmonic acid/ethylene, oxidative burst and camalexin related genes was assessed in Arabidopsis. T. atroviride induced an overlapped expression of defence-related genes of SA and JA/ET pathways, and of the gene involved in the synthesis of the antimicrobial phytoalexin, camalexin, both locally and systemically. This is the first report where colonization of Arabidopsis roots by T. atroviride induces the expression of SA and JA/ET pathways simultaneously to confer resistance against hemibiotrophic and necrotrophic phytopathogens. The beneficial effects induced by the inoculation of Arabidopsis roots with T. atroviride and the induction of the plant defence system suggest a molecular dialogue between these organisms.  相似文献   

5.
The effects of soil moisture (varying from 15% to 42% (v/v)) and sowing depth (1.5–6.0 cm) on the development of bean plants grown in sterile soil infested by the pathogen Rhizoctonia solani and its antagonist Trichoderma harzianum were studied under greenhouse conditions. The four possible combinations of soil infestation with both fungi were tested. Disease severity, percentage of plants emerged, plant height and dry weight were evaluated 3 weeks after sowing. Emergence rate and growth of plants inoculated only with R. solani were not affected by soil moisture, but in the presence of both fungi, plant emergence, plant height and dry weight significantly decreased when soil moisture diminished. Deep sowing significantly reduced the emergence rate and growth of those plants that were inoculated with R. solani only. However, when the soil was infested with both fungi, the effect of sowing depth was not significant. At a sowing depth of 6.0 cm, the percentage of plants emerged was 50% in the presence of T. harzianum, but only 6.7% when the pathogen was inoculated alone. The antagonist protected bean seedlings from pre-emergence damping-off, reduced disease severity and increased plant growth in the presence of R. solani, especially in moist soil.  相似文献   

6.
Trichoderma spp. are used as antagonists against different pathogens. Despite many possibilities of using Trichoderma as an antagonist, there are gaps in the knowledge of the interaction between Trichoderma, cassava and Scytalidium lignicola. This fungus causes cassava black root rot and is an inhabitant of the soil, so it is difficult to control. Antagonists may contribute to the possible induction of resistance of plants because, when exposed to such pathosystems, plants respond by producing antioxidative enzymes. The test for potential inhibition of growth of S. lignicola CMM 1098 in vitro was performed in potato-dextrose-agar with two Trichoderma strains T. harzianum URM3086 and T. aureoviride URM 5158. We evaluated the effect of the two selected Trichoderma to reduce the severity of cassava black root rot and shoots. Subsequently, the production of enzymes (ascorbate peroxidase, catalase, peroxidase and polyphenol oxidase) was evaluated in cassava plants. All two Trichoderma strains show an inhibition of the growth of S. lignicola CMM 1098. The most efficient was T. harzianum URM 3086, with 80.78% of mycelial growth inhibition. T. aureoviride URM 5158 was considered the best chitinase producer. All treatments were effective in reducing severity, especially treatments using Trichoderma. Cassava plants treated with T. aureoviride URM 5158 had the highest enzyme activity, especially peroxidase and ascorbate peroxidase. Trichoderma harzianum URM3086 and Trichoderma aureoviride URM 5158 were effective in reducing the severity of cassava black root rot caused by S. lignicola CMM 1098.  相似文献   

7.
The flight and parasitism capacities and emergence rate of fiveTrichogramma species [T. acacioi (Brun),T. bruni (Nagaraja),T. demoraesi (Nagaraja),T. maxacalii (Voegelé & Pointel) andT. soaresi (Nagaraja)] were compared. The flight capacity of these parasitoids was assessed in test units. A tube was placed inside each test unit with a piece of card with 300Anagasta kuehniella (Zeller) (Lepidoptera: Pyralidae) eggs parasitized by one of theTrichogramma species. The emerged adults inside the test units were classified according to their position as flyers, non-flyers and walkers. Cards withA. kuehniella eggs, parasitized by one of theTrichogramma species, were assessed to determine the parasitism rate and emergence of these natural enemies. The data were submitted to analysis of variance and the means compared by the Scott-Knott test at 5% probability.Trichogramma maxacalii exhibited better flight capacity than the other species, in addition to good parasitism rate and high percentage of emergence. http://www.phytoparasitica.org posting May 10, 2007.  相似文献   

8.
This work was undertaken to determine the effects ofTrichoderma spp. combined with label and sublabel rates of metam sodium on survival ofRhizoctonia solani in soil. Soils were infested with wheat bran preparations ofTrichoderma hamatum Tri-4,T. harzianum Th-58,T. virens Gl-3, andT. viride Ts-1-R3. Soil was also infested with sterile beet seeds that were colonized withR. solani. Beet seeds were later recovered, plated onto water agar plus antibiotics, and the growth ofR. solani was recorded. Preliminary experiments showed thatT. hamatum andT. virens reduced survival and saprophytic activity ofR. solani when the biocontrol fungi were incorporated into soil at 1.5% (w:w) or greater. Based on these data, biocontrol fungi in subsequent experiments were incorporated into soil at 2%. Metam sodium at label rate killed all biocontrol fungi andR. solani. At 1:2 and 1:5 dilutions, metam sodium reduced survival ofR. solani and allTrichoderma spp. When biocontrol fungi plus the label rate of metam sodium and 1:5, 1:10, 1:50 or 1:100 dilutions of the label rate were tested together, there were no interactions between any biocontrol agent and the fumigant with respect to colony diameter, reflecting that allTrichoderma isolates tested reacted similarly to increasing concentrations of metam sodium. At the label rate of metam sodium, allTrichoderma spp. significantly reduced colony diameter, but not growth rate, ofR. solani from beet seed. For the levels of metam sodium tested in combination withTrichoderma, it does not appear feasible to use a reduced rate of metam sodium to controlR. solani. However, the combination ofTrichoderma with metam sodium does reduce growth ofR. solani in comparison with that provided by metam sodium at the label rate. http://www.phytoparasitica.org posting Feb. 11, 2004.  相似文献   

9.
Since 2003, Torenia fournieri plants grown for experimental purposes were repeatedly infected by powdery mildew in a laboratory in Hungary. Based on morphological characteristics, the pathogen belonged to the mitosporic genus Oidium subgen. Reticuloidium, the anamorph stage of Golovinomyces. The rDNA ITS sequence was identical to that of two other powdery mildew fungi, infecting Arabidopsis and Veronica, respectively, in different parts of the world. According to a previous phylogenetic analysis of ITS and 28S rDNA sequences, those two powdery mildews belong to a recently evolved group of Golovinomyces characterized by multiple host range expansions during their evolution. Both the ITS sequence and the morphological data indicate that the powdery mildew anamorph infecting Torenia also belongs to this group. It is likely that the powdery mildew infections of the experimental T. fournieri plants, native to south-east Asia, were the result of a very recent host range expansion of a polyphagous Golovinomyces because (i) T. fournieri is absent from our region, except as an experimental plant grown in the laboratory, (ii) the powdery mildew fungus infecting this exotic plant belongs to a group of Golovinomyces where host range expansion is a frequent evolutionary scenario, (iii) cross-inoculation tests showed that this pathogen is also able to infect other plant species, notably A. thaliana and tobacco, and (iv) no Golovinomyces species are known to infect T. fournieri anywhere in the world. Although host range expansion has often been proposed as a common evolutionary process in the Erysiphales, and also in other biotrophic plant pathogens, this has not been clearly demonstrated in any case studies so far. To our knowledge, this is the first convincing case of a host range expansion event in the Erysiphales.  相似文献   

10.
Trichoderma harzianum is an effective biocontrol agent against the devastating plant pathogen Rhizoctonia solani. Despite its wide application in agriculture, the mechanisms of biocontrol are not yet fully understood. Mycoparasitism and antibiosis are suggested, but may not be sole cause of disease reduction. In the present study, we investigated the role of oxidant-antioxidant metabolites in the root apoplast of sunflower challenged by R. solani in the presence/absence of T. harzianum NBRI-1055. Analysis of oxidative stress response revealed a reduction in hydroxyl radical concentration (•OH; 3.6 times) at 9 days after pathogen inoculation (dapi), superoxide anion radical concentration (O2•−; 4.1 times) at 8 dapi and hydrogen peroxide concentration (H2O2; 2.7 times) levels at 7 dapi in plants treated with spent maize-cob formulation of T. harzianum NBRI-1055 (MCFT), as compared to pathogen-inoculated plants. The protection afforded by the biocontrol agent was associated with the accumulation of the ROS gene network: the catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and ascorbate peroxidase (APx), maximum activity of CAT (11.0 times) was observed at 8 dapi, SOD (7.0 times) at 7 dapi, GPx (5.4 times) and APx (8.1 times) at 7 dapi in MCFT-treated plants challenged with the pathogen. This was further supported by the inhibition of lipid and protein oxidation in Trichoderma-inoculated plants. MCFT stimulated the accumulation of secondary metabolites of phenolic nature that increased up to five-fold and also exhibited strong antioxidant activity at 8 dapi, eventually leading to the systemic accumulation of phytoalexins. These results suggest that T. harzianum–mediated biocontrol may be related to alleviating R. solani-induced oxidative stress.  相似文献   

11.
A ring spot disease of Aloe vera was found on leaves of potted seedlings of Aloe vera in Hachijojima and Chichijima Islands, Tokyo. From tissue of ring spot lesions, a fungus producing Fusarium-type conidia was consistently isolated. After 1 month, reddish perithecia of nectriaceous fungus had formed on the colonies of this isolate on PDA. These nectriaceous and Fusarium fungi were identified as Haematonectria haematococca and Fusarium sp., respectively. From a single ascospore isolation, the former was confirmed to be the teleomorph of the Fusarium sp. Typical ring spot lesions were reproduced by artificial inoculations using single ascospore and single conidium isolates. Inoculations of five species of genus Aloe revealed that they were highly susceptible except for A. arborescens. This is the first report of a disease on Aloe caused by H. haematococca (anamorph: Fusarium sp.) in Japan, and it was named aloe ring spot.  相似文献   

12.
Selective media without pentachloronitrobenzene were developed for quantitative assays of Fusarium oxysporum in soils. Media Fo-G1 and Fo-G2 were effective for naturally infested soils, Fo-W1 and Fo-W2 for wild-type isolates in soils containing a nitrate-nonutilizing (nit) mutant, and Fo-N1 and Fo-N2 for nit mutants. Selective media were made using ammonium citrate dibasic, l-sorbose, econazole nitrate, 25% iminoctadine triacetate solution and 50% tolclofos-methyl wettable powder for soil dilutions of 100-fold or more (Fo-G1, FoW1 and Fo-N1) and 10-fold (Fo-G2, Fo-W2 and Fo-N2). Potassium chlorate was added to Fo-N1 and Fo-N2. The efficacy for selectively isolating F. oxysporum was confirmed using six soils naturally infested with one of six formae speciales of F. oxysporum and with soil dilutions containing conidia of wild-type strains or nit mutants from the six formae speciales. On Fo-G1 and Fo-G2, most colonies of F. oxysporum were compact and round with purplish or reddish pigment in the reverse. Cylindrocarpon sp. formed colonies as large as those of F. oxysporum but were distinguishable by their colony morphology. Other contaminants such as F. solani, F. moniliforme, and Trichoderma were suppressed by medium ingredients and colonies of F. oxysporum. On Fo-W1 and Fo-W2, colony morphology of F. oxysporum and contaminants corresponded to that on Fo-G1 and Fo-G2, although F. oxysporum failed to produce the pigment. On Fo-N1 and Fo-N2, nit mutants formed clear colonies from 100- and 10-fold soil dilutions, respectively, and contaminants seldom formed large colonies.  相似文献   

13.
Native Trichoderma spp. were isolated from agricultural fields in several regions of Ecuador. These isolates were characterized via morphological observation as well as molecular phylogenetic analysis based on DNA sequences of the rDNA internal transcribed spacer region, elongation factor-1α gene and RNA polymerase subunit II gene. Fifteen native Trichoderma spp. were identified as T. harzianum, T. asperellum, T. virens and T. reesei. Some of these strains showed strong antagonistic activities against several important pathogens in Ecuador, such as Fusarium oxysporum f. sp. cubense (Panama disease) and Mycosphaerella fijiensis (black Sigatoka) on banana, as well as Moniliophthora roreri (frosty pod rot) and Moniliophthora perniciosa (witches’ broom disease) on cacao. The isolates also showed inhibitory effects on in vitro colony growth tests against Japanese isolates of Fusarium oxysporum f. sp. lycopersici, Alternaria alternata and Rosellinia necatrix. The native Trichoderma strains characterized here are potential biocontrol agents against important pathogens of banana and cacao in Ecuador.  相似文献   

14.
The development time and parasitization rate ofDiaeretiella rapae (M’Intosh) onBrevicoryne brassicae (L.) feeding on differentBrassica cultivars was studied in the laboratory at 20°C. The shortest development time from egg to adult parasitoid was 11.6 days on cabbage cv. ‘Yalova 1’ and the longest was 12.1 days on turnip cv. ‘Antep’ and rapeseed cv. local variety. Females lived significantly longer than males on the host plants used in the study. Females and males had the shortest longevity on rapeseed at 11.1 and 5.1 days, respectively. The highest percent parasitism ofB. brassicae byD. rapae was found on cabbage (40.20%), and the lowest was recorded on turnip (32.64%). Our results demonstrate that parasitism rate could be influenced by the plant quality, probably due to the nutritional status of the aphids or to toxic compounds ingested through the plant. Cabbage, cauliflower and broccoli were found to be suitable plants for the parasitoid, considering the development time of pre-adults, and the parasitization rate ofD. rapae onB. brassicae. http://www.phytoparasitica.org posting Jan. 23, 2007.  相似文献   

15.
Tomato chlorosis virus causes yellow leaf disorder epidemics in many countries worldwide. Plants of Physalis ixocarpa showing abnormal interveinal yellowing and plants of Physalis peruviana showing mild yellowing collected in the vicinity of tomato crops in Portugal were found naturally infected with ToCV. Physalis ixocarpa and P. peruviana were tested for susceptibility to ToCV by inoculation with Bemisia tabaci, Q biotype. Results confirmed that ToCV is readily transmissible to both species. The infection was expressed in P. ixocarpa by conspicuous interveinal yellow areas on leaves that developed into red or brown necrotic flecks, while P. peruviana test plants remained asymptomatic. Infected plants of both P. ixocarpa and P. peruviana served as ToCV sources for tomato infection via B. tabaci transmission. This is the first report of P. ixocarpa and P. peruviana as natural hosts of ToCV.  相似文献   

16.
Xanthomonas oryzae pv. oryzae strain T7174R is lysed by bacteriophage OP1h and OP1h2. Three mutants tolerant to both OP1h and OP1h2 were isolated by transposon mutagenesis. The mutants had an insertion of the transposon in XOO1687, which is predicted to encode a TonB-dependent receptor gene. Plasmid pHMIroNB that contained XOO1687 of T7174R was constructed, and the mutant was transformed with the plasmid. The transformant recovered sensitivity to OP1h and OP1h2. Electron microscopic analysis demonstrated that OP1h and OP1h2 can adsorb to the wild type and the transformant, but they could not adsorb to the phage-tolerant mutant. These results suggest that the TonB-dependent receptor gene relates to adsorption and infection of T7174R by OP1h2 and OP1h. Y. Inoue and S. Tsuge have contributed equally to this work.  相似文献   

17.
Twenty-eight isolates of Fusarium oxysporum f. sp. spinaciae (FOS; the causal agent of spinach wilt) collected from Japan were assessed for mating type and subjected to phylogenetic analysis. Mating type analysis revealed all isolates to be MAT1-2, suggesting that there is no sexual recombination within the population. Phylogenetic analyses based on nucleotide sequences of the ribosomal DNA intergenic spacer (IGS) and the mating type locus (MAT1) suggested that FOS is polyphyletic. The cluster analysis based on IGS showed four phylogenetic groups (S1–S4) among the isolates. Two distinct lineages, S1 and S3, included FOS isolates both of the vegetative compatibility group (VCG) types, 0330 and 0331, demonstrating that VCG differentiation in FOS may not necessarily reflect the phylogenetic relationships based on IGS and MAT1-2-1.  相似文献   

18.
 Green fluorescent protein (GFP)-marked Fusarium oxysporum f. sp. melonis and nonmarked F. oxysporum f. sp. fragariae were stained with neutral red. The neutral red stained vacuoles of the fungi without disturbing GFP fluorescence in the cytoplasm. GFP-marked fungi showed fluorescent hyphae with dark-stained vacuoles, whereas nonmarked fungi were detected as nonfluorescent hyphae with dark-dotted vacuoles. Root colonization by these two fungi was monitored using this method. Microconidia attached similarly to the root surface and elongated vegetative hyphae. Only the pathogenic fungi invaded, causing necrosis at the inoculation site. Thus, the present method enabled us to track simultaneously the various formae speciales of F. oxysporum colonizing the root surface. Received: March 25, 2002 / Accepted: September 27, 2002  相似文献   

19.
Antibiosis is assumed to be an essential mechanism exerted by potential biocontrol agents (BCAs) of Trichoderma spp. Therefore, in the present study, we report for the first time on the elucidation and production of viridiofungin A (VFA) from T. harzianum isolate T23 cultures and investigate the antifungal potential of VFA and some other secondary metabolites purified from T. harzianum cultures against Fusarium moniliforme. The bioautography assay revealed that T. harzianum isolates T16 and T23 excreted several secondary metabolites with antifungal activity. Following isolation and purification of the antifungal zones, three fractions (F223, F323 and F423) from extracts of isolate T23 and two fractions (F416 and F516) from extracts of isolate T16 exhibited pronounced fungitoxic activity in the bioautography and antibiotic disk assays against Cladosporium spp. and F. moniliforme, respectively. The structure of the antifungal metabolite in fraction F323 was identified as viridiofungin A (VFA), the first report of production of VFA by isolate T23 of T. harzianum. Following cultivation of isolate T23 in PDB medium for 9 days, 94.6 mg l−1 of VFA were determined. VFA and fraction F516 retarded the mycelial growth of F. moniliforme in the non-volatile phase assay by >90% for each 250 μg ml−1 7 days post-inoculation (dpi). While VFA and fraction F416 showed both volatile and non-volatile effects, fraction F516 seemed to exhibit mainly non-volatile activity. Microscopic examination revealed that hyphae of F. moniliforme grown on VFA-amended medium were less branched and appeared thicker than untreated hyphae. Furthermore, in the presence of VFA, formation of chlamydospores by F. moniliforme was increased. Finally, the antifungal spectrum of VFA towards various important plant pathogens was evaluated. Germination of propagules of a variety of fungal pathogens in vitro was differentially inhibited by VFA. While in the presence of 100 μg ml−1 VFA conidial germination of V. dahliae was completely inhibited, a slightly higher concentration (150 μg ml−1) of the inhibitor was required to suppress germination of Phytophthora infestans sporangia or sclerotia of Sclerotinia sclerotiorum. Contrary to several reports in the literature, VFA proved to be fungistatic rather than fungicidal. However, neither VFA nor the other Trichoderma metabolites, such as 6PAP, F416 and F516, exhibited any antibacterial activity against Gram-positive and Gram-negative bacteria.  相似文献   

20.
 In May 1998 leaf spot caused by Septoria chrysanthemella was found on Aster savatieri in Kanagawa Prefecture, Japan. This is the first report of leaf spot on A. savatieri caused by S. chrysanthemella. Received: September 13, 2002 / Accepted: October 18, 2002 Acknowledgments The authors thank Dr. T. Kobayashi, formerly of Tokyo University of Agriculture, for his advice on identifying the fungus.  相似文献   

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