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1.
Scanning electron microscopy showed that hyphae of Coniothyrium minitans produced appressorium-like swellings when they came in contact with Sclerotinia sclerotiorum in dual culture on PDA. The parasitized hyphae gradually skrank and collapsed, and hyphae of the mycoparasite were found inside the host hyphae. The mycoparasite hyphae grew inter- and intracellularly within the sclerotia of S. sclerotiorum. In the later stages of parasitism, hyphae of the mycoparasite proliferated extensively within the sclerotia and formed pycnidia near the sclerotial surface. At this stage, the sclerotia became flattened, soft and disintegrated. Sclerotia parasitized by C. minitans failed to germinate either myceliogenically or carpogenically.  相似文献   

2.
The effect of pollen and senescent petals on the suppression of alfalfa (Medicago sativa L.) blossom blight (Sclerotinia sclerotiorum) by the mycoparasite Coniothyrium minitans was investigated. When incubated at 20°C for 39 h, germination of conidia of C. minitans and ascospores of S. sclerotiorum was 99.9 and 98.6%, respectively, in the presence of alfalfa pollen (9×104 pollen grains mL?1), whereas spore germination of both organisms was <0.5% in the absence of pollen (in water). In the presence of a commercial pollen product, Swiss? pollen granules (mainly bee pollen), germination was 99.6% for C. minitans and 98.3% for S. sclerotiorum when the pollen concentration was 1.0% (w/v). When the pollen concentration was reduced to 0.1% (w/v), germination was reduced to 13.0% for C. minitans and 10.8% for S. sclerotiorum. Tests on detached alfalfa florets showed that the colonization of alfalfa florets by S. sclerotiorum was significantly suppressed by C. minitans in the presence of pollen (1.0% Swiss? pollen granules), especially when C. minitans was inoculated 1-day before S. sclerotiorum. In vivo inoculation tests revealed that the efficacy of C. minitans in the protection of alfalfa pods from the infection by S. sclerotiorum was affected by the time at which C. minitans was applied. When C. minitans was applied on young blossoms of alfalfa at the anthesis stage, pod infection was 96.6% for the treatment of C. minitans+S. sclerotiorum and 99.6% for the treatment of S. sclerotiorum alone. However, when C. minitans was applied on senescent petals of alfalfa at the pod development stage, pod infection was 8.0% for the treatment of C. minitans+S. sclerotiorum compared to 90.8% for the treatment of S. sclerotiorum alone. These results suggest that timing of the application of C. minitans is critical for the mycoparasite to compete with S. sclerotiorum for the source of nutrients from pollen and senescent petals, and for its control of alfalfa blossom blight caused by S. sclerotiorum.  相似文献   

3.
A glasshouse and field trial were conducted to evaluate foliar sprays of Contans® WG (Coniothyrium minitans) conidial suspensions for control of sclerotinia rot of carrot and infection of Sclerotinia sclerotiorum sclerotia by C. minitans. In the glasshouse trial, foliar sprays (1×104–108 conidia mL?1) decreased the viability of sclerotia recovered from diseased plants and increased infection by C. minitans. In the field trial, three successive foliar sprays applied at 14-day intervals failed to reduce foliage disease severity, but significantly reduced viability of sclerotia recovered from diseased plants/crop debris and increased infection by C. minitans. No significant differences in sclerotial viability or infection were observed between two conidial concentrations (2.4 and 4.8×106 conidia mL?1). Foliar sprays of Contans® WG have potential for reducing viability of sclerotia produced on diseased foliage.  相似文献   

4.
Three strains of Sclerotinia sclerotiorum, namely Ep-1PB (PB), Ep-1PK (PK) and Ep-1PNA5 (A5), were compared for the production of oxalic acid (OA) on potato dextrose agar (PDA) and Maxwell agar medium (MAM) and for mycelial susceptibility to infection by the mycoparasite Coniothyrium minitans on PDA. Results showed that strain PB produced negligible oxalate, whereas strain PK was detected to produce oxalate, but much less than that produced by strain A5. The three investigated strains differed slightly in mycelial growth rates and mycelial biomass on PDA. However, colonies of strains PB and PK formed on PDA were more susceptible to invasion by C. minitans than colonies of strain A5. Meanwhile, amendment of synthetic oxalate in PDA at 0.25–2.00 mg g−1 medium suppressed aggressiveness of C. minitans in invasion of colonies of S. sclerotiorum strain PB developed on this medium. These results suggest that infection of hyphae of S. sclerotiorum is negatively affected by the presence of oxalate. The importance of oxalate degradation by C. minitans in its mycoparasitism on hyphae of S. sclerotiorum provides a clue for improvement of the biocontrol efficacy of C. minitans in the future.  相似文献   

5.
Biological control agents (BCAs) Bacillus subtilis QST 713, Coniothyrium minitans CON/M/91-08, Streptomyces lydicus WYEC 108, and Trichoderma harzianum T-22 were evaluated for their efficacy in the reduction of survival of sclerotia and production of apothecia of Sclerotinia sclerotiorum under controlled environments. A growth chamber assay was conducted where 25 sclerotia were buried in pots containing potting soil, and BCAs were drenched into the soil at various concentrations, and five soybean seeds were planted in each pot. The presence and number of S. sclerotiorum apothecia were recorded daily. Sclerotinia sclerotiorum sclerotia were retrieved six weeks after seeding and viability was assessed on water agar plates. All BCAs were effective in reducing S. sclerotiorum inoculum at various efficacies. In general, efficacy was positively correlated with the rate of application. At the rate of application when the efficacy did not change significantly by increasing the rate, the BCAs had various reductions of apothecia and sclerotia. B. subtilis reduced apothecia and sclerotia by 91.2 and 29.6%, respectively; C. minitans reduced apothecia and sclerotia by 81.2 and 50%, respectively; Streptomyces lydicus reduced apothecia and sclerotia by 100 and 29.6%, respectively; Trichoderma harzianum reduced apothecia and sclerotia by 80.5 and 31.7%, respectively. In addition, the commercial strain of C. minitans CON/M/91-08, and a wild Michigan strain of C. minitans W09 were compared for their growth and sclerotial reduction. W09 had faster growth rate than the commercial strain, indicating potential diversities of biological control strains to be studied.  相似文献   

6.
The relatively slow germination rate of Coniothyrium minitans limits its control efficiency against Sclerotinia sclerotiorum. Pre-germinated conidia of C. minitans enhanced its efficiency significantly: in foliar experiments with oilseed rape, hyphal extension of S. sclerotiorum was inhibited by 68%, while formation of sclerotia was completely inhibited when pre-germinated conidia were applied.Revisions requested 27 July 2004; Revisions received 7 September  相似文献   

7.
Folsomia Candida was maintained on potato dextrose agar (PDA) plates precolonised by the mycoparasite Coniothyrium minitans for 3 yr but the sciarid Bradysia sp. survived for a maximum of only three generations. Collembolans and sciarid larvae from these cultures were able to transmit C. minitans to uninoculated PDA plates through the survival of spores in faecal pellets. Adult and larval sciarids also transmitted C. minitans from PDA culture to uninoculated PDA plates by contamination on the cuticle. In soil and sand both sciarids and collembolans were able to transmit C. minitans from C. m/m'tans-inoculated to uninoculated sclerotia of Sclerotinia sclerotiorum. Inoculation of sclerotia with C. minitans enabled greater populations of larger collembolans to develop. In the glasshouse where C. minitans had been applied to the soil, one adult sciarid and four collembolans out of 70 and 101 insects collected respectively yielded C. minitans on placement onto PDA + Aureomycin.  相似文献   

8.
9.
Biological control agents (BCAs) were evaluated for their efficacy on reducing the number of sclerotia of Sclerotinia sclerotiorum (Lib.) de Bary in the soil and on Sclerotinia stem rot in soybean production systems in Michigan. BCAs included Coniothyrium minitans CON/M/91–08 (Product name: Contans®WG), Streptomyces lydicus WYEC 108 (Actinovate®AG), Trichoderma harzianum T-22 (PlantShield®HC), and Bacillus subtilis QST 713 (Serenade®MAX). At two field locations, soil artificially infested with S. sclerotiorum sclerotia, was treated by incorporating the above BCAs in the topsoil before planting and boscalid was applied as a foliar fungicide at growth stage R1 as a positive control. C. minitans was the most effective BCA and reduced the disease severity index (DSI) by 68.5% and the number of sclerotia of S. sclerotiorum in the soil by 95.3%. S. lydicus and T. harzianum reduced DSI by 43.1% and 38.5% and sclerotia in soil by 90.6% and 70.8%, respectively. B. subtilis only had a marginal effect on S. sclerotiorum. Populations of Bacillus, Streptomyces, Trichoderma spp., and C. minitans collected from soil samples and at 3, 28, 71, and 169 days after BCA application indicated that the population of Streptomyces, Trichoderma spp., and C. minitans did not change significantly throughout the season, which may be the reason for their effectiveness.  相似文献   

10.
A study was conducted to determine water-assisted dissemination of conidia of Coniothyrium minitans (Cm), a mycoparasite of Sclerotinia sclerotiorum (Ss), in four soils (yellow–brown soil, red-clay soil, fluvo-aquic soil and black soil) and one sand. Conidial suspensions (1×107 conidia mL?1) of Cm were applied to sieved (2 mm screen) soil or sand in glass tubes to test vertical dissemination (VD) and in aluminum boxes to test horizontal dissemination (HD) of conidia. Results showed that conidia of Cm could be disseminated with water and spread in soil or sand for 16–20 cm vertically and for 5–10 cm horizontally. The conidial concentration of Cm was logarithmically reduced with the increase in depth of VD or the distance of HD. Dissemination of Cm conidia in sand was better than that in four soils. Potting experiments were done to further understand the potential of water-assisted dissemination of Cm conidia in suppression of Ss carpogenic germination. Results showed that more apothecia were produced by Ss sclerotia located at the soil surface than those at 5 and 10 cm in depth. The minimum Cm concentration for suppression of Ss carpogenic germination was 1000 conidia g?1 soil. Two-season field trials indicated that water-assisted application of Cm was an effective strategy used at the time for transplanting oilseed rape seedlings to suppress Ss carpogenic germination, thereby reducing the primary infection source for sclerotinia diseases of oilseed rape.  相似文献   

11.
Nine fungal isolates [Clonostachys rosea (1), Coniothyrium minitans (1), Trichoderma crassum (1), T. hamatum (4), T. rossicum (1) and T. virens (1)] were tested in two bioassays for their ability to degrade sclerotia and reduce apothecial production and carpogenic infection of cabbage seedlings. C. minitans LU112 reduced apothecial production in both bioassays, with T. virens LU556 significantly reducing apothecial production in the sclerotial parasitism assay. Both isolates also reduced sclerotial viability in this assay to 5% for C. minitans and 22% for T. virens. C. minitans LU112 and T. virens LU556 reduced the infection of cabbage seedlings in the pot bioassay 126 days after sowing but not after 147 days, partly due to ascospore cross-infection between treatments. C. minitans LU112, T. virens LU556 and T. hamatum LU593 as maizemeal-perlite (MP) soil incorporation and transplant potting mix incorporation were evaluated for their ability to control Sclerotinia sclerotiorum disease of cabbage in field experiments. S. sclerotiorum infection of cabbage was reduced by 46–52% and 31–57% by both C. minitans LU112 and T. hamatum LU593 as MP soil incorporations, respectively, in the two field experiments. T. virens LU556 MP soil incorporation and C. minitans LU112 and T. hamatum LU593 transplant potting mix incorporations reduced S. sclerotiorum disease in the first experiment but not in the second experiment. A commercial C. minitans LU112 formulation, C. Mins LU112 WG, also significantly reduced S. sclerotiorum disease by 59%. Soil incorporation of C. minitans and T. hamatum was shown to have potential to control S. sclerotiorum disease in cabbage.  相似文献   

12.
Field experiments were conducted during 1992–1994 to evaluate the effectiveness of five indigenous fungi for control of white mold (Sclerotinia sclerotiorum) of dry bean (Phaseolus vulgaris). The five fungi consisted of one antagonist, Epicoccum purpurascens, and four mycoparasites, Coniothyrium minitans, Talaromyces flavus, Trichothecium roseum, and Trichoderma virens. Spore suspensions of each fungus were sprayed onto bean plants two or three times during the early bloom to midbloom period. Incidence of white mold of dry bean was significantly reduced by all biocontrol agents. C. minitans and E. purpurascens, the most effective agents, reduced the proportion of plants infected by an average of 56 and 43%, respectively (P < 0.001). C. minitans was the only biocontrol agent recovered consistently from sclerotia and diseased seed present in harvested samples. It was recovered at similar frequencies in samples from all treatments. Of the sclerotia of S. sclerotiorum collected from harvested seed, 59% were infected by C. minitans in 1993 and 20% were infected by C. minitans in 1994. In three additional trials in 1994, comparing C. minitans with the fungicide benomyl, the fungus was not effective in any of the experiments, whereas benomyl reduced disease incidence relative to the control in one trial. The study suggests that, among the five indigenous fungi, C. minitans is the most promising agent for control of white mold of dry bean under Canadian prairie conditions.  相似文献   

13.
J. Kaur    G. D. Munshi    R. S. Singh    E. Koch 《Journal of Phytopathology》2005,153(5):274-279
Three isolates of Trichoderma atroviride and two isolates of Coniothyrium minitans known to efficiently degrade sclerotia of Sclerotinia sclerotiorum were cultured on minimal medium with sucrose, carboxymethyl cellulose (CMC), xylan, laminarin, colloidal chitin or powdered sclerotia as carbon source. The activity of endochitinase, endo‐β‐1,3‐glucanase, endoxylanase and endocellulase in culture filtrates was determined after 7 and 15 days of culture using dye‐labelled substrates. The strongest inducers of chitinase were colloidal chitin and sclerotia powder. Chitinase activity appeared to be faster induced in the isolates of T. atroviride than in the isolates of C. minitans, but the maximum level of activity present in culture filtrates of the two species was similar. With CMC and xylan as carbon source, concurrent production of the corresponding enzymes was observed for the Trichoderma isolates. The isolates of C. minitans produced cellulase on xylan but not on CMC, whereas xylanase was produced on both carbon sources. Laminarin induced the formation of glucanases in the three isolates of T. atroviride but not the isolates of C. minitans. However, in the sclerotia‐containing cultures of C. minitans glucanase activity was even higher than in the respective cultures of the Trichoderma isolates. During the 31‐day study period, the pattern of enzyme production in shake cultures containing sclerotia powder was very similar for the isolates of T. atroviride and C. minitans. Glucanase activity generally reached a peak 24 days after inoculation of the flasks, whereas the activity of chitinase, cellulase and xylanase remained fairly constant throughout the experiment.  相似文献   

14.
White mold is a major disease in commercial soybean production. An effective measure to reduce the negative effects of Sclerotinia sclerotiorum is the use of bio-fungicides. Strains of Coniothyrium minitans were isolated and efficacy tests against S. sclerotiorum was studied. The efficacy of pycnidiospores sprays of strain N09 (GenBank Accession No HQ908274) from Iowa, USA and strain CON/M/91-08 of Contans® WG were compared in a series of experiments. Sclerotia viability was significantly (P < 0.05) lower in both sclerotia-infested-sterilized-soils (SISS) and sclerotia-infested-unsterilized-soils (SIUS) sprayed with N09 compared with CON/M/91-08 and control at 3°C for 75d and 90d sampling. Similarly, sclerotia viability was significantly (P < 0.05) lower at 23°C for 45, 60 and 75d sampling in SISS and 45, 75 and 90 d sampling in SIUS compared with CON/M/91-08 and control. In contrast, viability of N09 colonies were significantly (P < 0.05) higher than that of CON/M/91-08 both at 3°C and 23°C in SISS across sampling periods. While in SIUS, N09 colonies were significantly higher at 3°C for 15, 30, 45, 75 and 90 d sampling, and at 23°C for 30, 60 and 75 d sampling. Also, (1) N09 had a faster growth rate and produced 1.5 times more pycnidiospores than CON/M/91-08; (2) mycoparasitism by N09 was faster than CON/M/91-08; and (3) co-inoculation of sclerotia and the strains, N09 showed lower sclerotia reproduction than CON/M/91-08. Our data suggest that the new strain N09 has a greater efficiency than CON/M/91-08 in killing sclerotia.  相似文献   

15.
Aims: Assessment of antimicrobial activity of the mycoparasite Coniothyrium minitans and its macrolide antibiotic macrosphelide A. Methods and Results: Thirteen isolates of C. minitans were tested for ability to inhibit a number of filamentous fungi, yeasts, oomycetes and bacteria in agar based tests. Activity was found against some ascomycetes, basidiomycetes, oomycetes and Gram‐positive bacteria, but not against zygomycetes, yeasts or Gram‐negative bacteria tested. Six C. minitans isolates (Conio, Contans, IVT1, CM/AP/3118, B279/1, A1/327/1) were found to produce macrosphelide A in liquid culture and no other antibiotics were detected. On agar, macrosphelide A inhibited growth of some ascomycetes, basidiomycetes, oomycetes and all four Gram‐positive bacteria tested, including the medically important Staphylococcus aureus with a minimum inhibitory concentration of ≤500 μg ml?1. There was no inhibition observed against the yeasts and Gram‐negative bacteria when macrosphelide A was tested at 700 μg ml?1. Conclusions: The spectrum and level of activity of macrosphelide A produced by C. minitans against micro‐organisms are extended markedly compared to previous reports. Significance and Impact of the Study: Macrosphelide A was effective against Staph. aureus. Further study on the control of this bacterium is merited in view of the development of antibiotic resistance.  相似文献   

16.
The effects of temperature and pH on growth and antibiotic production by three isolates of Coniothyrium minitans (Conio, Contans and IVT1), known to produce the macrolide antibiotic macrosphelide A, were examined in modified Czapek Dox broth (MCD). Antibiotic production was determined by incorporating heated (60°C for 5 min) C. minitans spent culture filtrates of MCD (10%, v/v) into potato dextrose broth and assessing the ability of the filtrates to inhibit growth of S. sclerotiorum. All isolates grew over the temperature range of 10–30°C, with the optimum at approximately 15–20°C. Antibiotics were produced by all isolates at 10–30°C. Culture filtrates of MCD from all isolates incorporated into PDB inhibited growth of S. sclerotiorum by >50%, whereas there was a reduction in inhibition at 30°C for Conio and IVT1 but not Contans. All three isolates grew over the pH range of 3–7, with greater biomass production in buffered pH 3–5 than the unbuffered control (pH 4.8) media. Antibiotics were produced by all isolates at pH 3–5. Culture filtrates of MCD from all three isolates grown at pH 3–5 inhibited growth of S. sclerotiorum, with the greatest effect on inhibition observed at pH 3. There were no differences in growth inhibition between isolates at pH 3 and 4, but culture filtrates from Conio grown at pH 5 inhibited S. sclerotiorum more than those of IVT1 grown at the same pH. The significance of these results for biocontrol and optimizing antibiotic production by C. minitans is discussed.  相似文献   

17.
Coniothyrium minitans is a sclerotial parasite of the plant-pathogenic fungus Sclerotinia sclerotiorum, and conidial production and parasitism are two important aspects for commercialization of this biological control agent. To understand the mechanism of conidiation and parasitism at the molecular level, we constructed a transfer DNA (tDNA) insertional library with the wild-type strain ZS-1. A conidiation-deficient mutant, ZS-1TN22803, was uncovered through screening of this library. This mutant could produce pycnidia on potato dextrose agar (PDA), but most were immature and did not bear conidia. Moreover, this mutant lost the ability to parasitize or rot the sclerotia of S. sclerotiorum. Analysis of the tDNA flanking sequences revealed that a peroxisome biogenesis factor 6 (PEX6) homolog of Saccharomyces cerevisiae, named CmPEX6, was disrupted by the tDNA insertion in this mutant. Targeted gene replacement and gene complementation tests confirmed that a null mutation of CmPEX6 was responsible for the phenotype of ZS-1TN22803. Further analysis showed that both ZS-1TN22803 and the targeted replacement mutants could not grow on PDA medium containing oleic acid, and they produced much less nitric oxide (NO) and hydrogen peroxide (H2O2) than wild-type strain ZS-1. The conidiation of ZS-1TN22803 was partially restored by adding acetyl-CoA or glyoxylic acid to the growth media. Our results suggest that fatty acid β-oxidation, reactive oxygen and nitrogen species, and possibly other unknown pathways in peroxisomes are involved in conidiation and parasitism by C. minitans.  相似文献   

18.
Aphelenchoides saprophilus nematodes fed on sclerotia, mycelium, and alginate-formulated pellets of Sclerotinia sclerotiorum, mycelium of Trichoderma harzianum, and mixed fungal cultures. As many as 500 nematodes were found inside individual sclerotia. Results suggest potential impacts of fungivory on S. sclerotiorum and its ecological interactions with plant hosts and biocontrol fungi.  相似文献   

19.
At the end of the spring 1987 growing season, the mycoparasite Sporidesmium sclerotivorum was applied at 0, 0.2, 2 or 20 kg ha‐1 to lettuce plants infected with Sclerotinia minor. Disease incidence was monitored in the same plots for five subsequent crops (three fall and two spring crops) without additional application of either pathogen or mycoparasite. Logistic growth curves were fitted to the data to describe disease progression over time for each inoculum level within each of the five crops. Within each crop, increasing the quantity of mycoparasite inoculum resulted in positive horizontal displacement of the curve with respect to time. As quantities of inoculum of S. sclerotivorum increased, inflection points of the disease progress curves increased at a decreasing rate. Thus, additional mycoparasite inoculum resulted in ever‐smaller increases in inflection point, and after a certain threshold level of mycoparasite inoculum (< 0.2 kg ha‐1), increases in inflection point did not result in meaningful increases in harvestable lettuce. Maximum rates of disease increase were not different among the treatments within each crop, but were different between crops. Maximum rates of disease increase averaged 3.4, 3.4, 2.1, 3.6 and 1.5% day‐1 for the fall 1987, spring 1988, fall 1988, spring 1989, and fall 1989, respectively. At all inoculum levels, the fall epidemics began later after planting than the spring epidemics.  相似文献   

20.
The fungus Sclerotinia sclerotiorum is a necrotrophic plant pathogen causing significant damage on a broad range of crops. This fungus produces sclerotia that serve as the long‐term survival structures in the life cycle and the primary inoculum in the disease cycle. Melanin plays an important role in protecting mycelia and sclerotia from ultraviolet radiation and other adverse environmental conditions. In this study, two genes, SCD1 encoding a scytalone dehydratase and THR1 encoding a trihydroxynaphthalene reductase, were disrupted by target gene replacement, and their roles in mycelial growth, sclerotial development and fungal pathogenicity were investigated. Phylogenetic analyses indicated that the deduced amino acid sequences of SCD1 and THR1 were similar to the orthologues of Botrytis cinerea. Expression of SCD1 was at higher levels in sclerotia relative to mycelia. THR1 was expressed at similar levels in mycelia and sclerotia at early stages, but was up‐regulated in sclerotia at the maturation stage. Disruption of SCD1 or THR1 did not change the pathogenicity of the fungus, but resulted in slower radial growth, less biomass, wider angled hyphal branches, impaired sclerotial development and decreased resistance to ultraviolet light.  相似文献   

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