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1.
【背景】菌核病是北细辛根部主要病害之一,木霉菌作为目前应用最广泛的生物防治真菌,利用木霉菌防治北细辛菌核病是目前研究的热点。【目的】通过稀释分离法对健康北细辛植株根际土壤进行菌株分离,以期筛选出有效拮抗北细辛菌核病的生防木霉菌。【方法】以北细辛菌核病菌为靶标菌,采用平板对峙培养、挥发性与非挥发性物质抑菌的方法对分离得到的木霉菌进行筛选,采用生长速率法对筛选出的木霉菌的发酵液进行抑菌效果测定,并采用硫代巴比妥酸法测定筛选出的木霉对北细辛菌核病菌的丙二醛(Malondialdehyde,MDA)含量、紫外吸收法测定过氧化氢酶(Catalase,CAT)活性、氮蓝四唑法测定超氧化物歧化酶(Superoxide Dismutase,SOD)活性、愈创木酚法测定过氧化物酶(Peroxidase,POD)活性的影响。【结果】从土壤中分离出木霉菌共14株,通过形态学和ITS-RPB2双基因联合构建系统发育树,鉴定其为哈茨木霉(Trichoderma harzianum)、钩状木霉(Trichoderma hamatum)、拟康氏木霉(Trichoderma koningiopsis)、深绿木霉(Tr...  相似文献   

2.
Randomly amplified polymorphic DNA (RAPD) analysis and the PCR assay were used in combination with dilution plating on a semiselective medium to detect and enumerate propagules of Trichoderma hamatum 382, a biocontrol agent utilized in compost-amended mixes. Distinct and reproducible fingerprints were obtained upon amplification of purified genomic DNA of T. hamatum 382 with the random primers OPE-16, OPH-19, and OPH-20. Three amplified DNA fragments of 0.35 (OPE-160.35), 0.6 (OPH-190.6), and 0.65 (OPH-200.65) kb were diagnostic for T. hamatum 382, clearly distinguishing it from 53 isolates of four other Trichoderma spp. tested. Some isolates of T. hamatum shared these low-molecular-weight fragments with T. hamatum 382. However, RAPD analysis of isolates of T. hamatum with all three random primers used in consecutive PCR tests distinguished T. hamatum 382 from other isolates of T. hamatum. These three RAPD amplicons were cloned and sequenced, and pairs of oligonucleotide primers for each cloned fragment were designed. Use of the primers in the PCR assay resulted in the amplification of DNA fragments of the same size as the cloned RAPD fragments from genomic DNA of T. hamatum 382. A combination of dilution plating on a semiselective medium for Trichoderma spp. and PCR, with the RAPD primers OPH-19, OPE-16, and OPH-20 or the three sequence-characterized primers, was used successfully to verify the presence of T. hamatum 382 propagules in nine different soil, compost, and potting mix samples. All 23 Trichoderma isolates recovered on semiselective medium from commercial potting mixes fortified with T. hamatum 382 were identified as T. hamatum 382, whereas 274 Trichoderma isolates recovered from the other nine samples were negative in the PCR assay. Thus, this highly specific combination of techniques allowed detection and enumeration of propagules of T. hamatum 382 in fortified compost-amended potting mixes. Sequence-characterized amplified region markers also facilitated the development of a very simple procedure to amplify DNA of T. hamatum 382 directly from fortified compost-amended potting mixes.  相似文献   

3.
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   

4.
Trichoderma hamatum, T. harzianum andT. koningii were isolated from wheat and rye-grass roots from a field in Western Australia. Frequency of occurrence ofTrichoderma spp. was higher on roots subjected to washing only, for both wheat and rye-grass than the roots which were surface-sterilized with 0.6% or 1.25% NaOCl.Trichoderma spp. were recovered at a higher frequency on PDA amended with lactic acid (pH 4.5) than on PDA alone (pH 5.6) or PDA with streptomycin. In general,Trichoderma spp. were isolated at a higher frequency from roots of wheat than that of rye-grass.T. hamatum occurred at a higher frequency in rye-grass roots than in wheat, whereasT. harzianum was more common in roots of wheat than in rye-grass, especially in seedling and milky ripe stages.T. koningii was recovered at a higher frequency from roots at seedling stage of rye-grass than wheat, the reverse being true at tillering stage.T. koningii was not recovered from roots of either host in any sampling when they were surface sterilized with 1.25% NaOCl.The take-all fungus was isolated from wheat and rye-grass roots more frequently at tillering and stem extension stages than others. It was severely pathogenic to both hosts in sterilized and non-sterilized soil.Addition of lactic acid, HCl or streptomycin to PDA did not affect the growth of theTrichoderma spp. tested, but the growth was slower on Martin's medium than on other media. In generalT. harzianum andT. koningii grow faster thanT. hamatum. The growth of the three species were not different at 20 and 25°C, but at 15°c growing of all species was significantly reduced.Incorporation of lactic acid into PDA prevented the bacterial growth in all treatments. Streptomycin too reduced but to a lesser degree than lactic acid. Surface sterilization with NaOCl decreased the recovery of both bacteria and fungi. T. hamatum andT. koningii reduced the mortality of wheat and rye-grass plants inoculated with the take-all fungus in sterilized and non-sterilized soil, whereT. harzianum did not protect wheat or rye-grass from infection by the take-all fungus.  相似文献   

5.
Three species of fungi namely Fusarium roseum USDB 0005, Curvularia lunata var. aeria USDB 0006 and Trichoderma hamatum USDB 0008, and a sterile isolate were found growing on wood shavings. Both F. roseum USDB 0005 and C. lunata var. aeria USDB 0006 and the sterile isolate were weakly cellulolytic while T. hamatum USDB 0008 was strongly cellulolytic. Enzyme assays showed that T. hamatum USDB 0008 produced all the three components (exoglucanase, endoglucanase and B-glucosidase) of the cellulase complex. The enzymatic activity of this strain is compared to that of cellulolytic strains isolated from other wood sources.  相似文献   

6.
Fungal isolates, with known activity against Sclerotinia spp. in laboratory assays, were tested for their ability to control Sclerotinia minor in four field experiments (1998–2000). In the first experiment, eight fungal isolates (Trichoderma hamatum LU595, LU593, LU592, Trichoderma virens LU555 and LU556, Coniothyrium minitans LU112, Clonostachys rosea LU115 and Trichoderma rossicum LU596) were evaluated by incorporating spore suspensions into transplant potting mix and planting lettuce seedlings into a S. minor infested field site. At harvest, Trichoderma hamatum LU595, LU593, T. virens LU555 and C. minitans LU112 reduced disease by 30–50% compared with the untreated control under very high disease pressure (100%). In further field experiments C. minitans LU112 and T. hamatum LU593, applied as maizemeal–perlite soil amendments or incorporated into the potting mix, reduced S. minor disease over a range of disease pressures (29–91%). Disease control was equivalent or greater than that achieved with the standard carbendazim fungicide treatment. Both isolates were shown to effectively colonize the lettuce rhizosphere and surrounding soil and this colonization may have protected the roots from infection by S. minor. Multiple applications of C. minitans LU112 or T. hamatum LU593 formulations gave no added disease control compared with a single application at planting. Commercial formulations of both C. minitans LU112 and T. hamatum LU593 applied as transplant treatments, solid substrate soil amendments or as a spore drench gave consistent disease control and are currently being developed further.  相似文献   

7.
Biological control of wilt of egg plant (Solanum melongena L.) caused by Fusarium solani was made with the application of five Trichoderma species, T. harzianum, T. viride, T. lignorum, T. hamatum and T. reesei. The effect of volatile and non-volatile antibiotics of Trichoderma origin on growth inhibition of the wilt pathogen was studied. T. harzianum showed maximum growth inhibition (86.44 %) of the pathogen through mycoparasitism. The non-volatiles produced by the Trichoderma species exhibited 100 % growth inhibition of the pathogen under in vitro condition. Production of siderophores and fungal cell wall degrading enzymes, chitinase and β-1,3-glucanase were found. Treatments with two most efficient Trichoderma species, T. harzianum and T. viride resulted in the decreasing population of Fusarium solani in soil thereby deterring disease incidence in field condition.  相似文献   

8.
Three fungi, isolated from soil from which Phytophthora was not obtained, were evaluated for antagonism of Phytophthora spp. shown to cause root rot of chestnut in South Australia. Trichoderma hamatum and T. pseudokoningii appeared to inhibit P. cinnamomi by mycoparasitism. with evidence of parallel growth and coiling, and both Trichoderma spp. and Gliocladium virens grew over P. cinnamomi in vitro, preventing further growth of this pathogen. Antibiotics produced by young T. hamatum cultures and G. virens in culture filtrate experiments inhibited growth of P. cinnamomi and P. citricola. with filtrate from 4-day-old cultures of G. virens showing the greatest potential for biocontrol. All three antagonists prevented P. cinnamomi and P. citricola from causing infection symptoms on micropropagated shoots of chestnut cvs Goldsworthy and Buffalo Queen in an in vitro excised shoot bioassay for biocontrol.  相似文献   

9.
Bip T containing about 109 spores of Trichoderma viride, applied to peat 10 days before inoculation of substrate with Phytophthora cryptogea, effectively controlled Phytophthora foot rot of gerbera. The biocide in dosage of 300 and 600 g/m3 inhibited the development of the pathogen in substrate. The other potential antagonists T. hamatum and T. viride applied into peat at a dosage of 600 g/m3, decreased Phytophthora foot rot development.  相似文献   

10.
The mode of hyphal interaction and parasitism ofPythium spp. andRhizoctonia solani byTrichoderma hamatum was studied by both phase-contrast and Nomarski differential interference-contrast microscopy. Directed growth of the mycoparasite toward its host was observed. In the area of interaction,T. hamatum produced appressorial-like structures attached to the host cell wall. Subsequently, several different types of interactions occurred.T. hamatum either grew parallel to and along the host hypha or coiled around its host. In the contrast regions the parasite formed bulbular or hook-like structures that contained granular cytoplasm. In other cases the parasite penetrated into and grew within the mycelium ofR. solani orP. ultimum. As a consequence of the attack, the host hypha became vacuolated, shrank, collapsed, and finally disintegrated. These observations suggest the involvement of parasitism followed by lysis rather than involvement of antibiotics in this host-mycoparasite relationship.  相似文献   

11.
Summary Coating cotton seeds withTrichoderma spp. reduced the incidence of disease caused byR. solani by up to 83% in the greenhouse.T. hamatum was more efficient in disease control at 20°C, whileT. harzianum was superior at 27°C. Disease severity was reduced by 47–60% in two field experiments, showing no statistical difference from treatment with pentachloronitrobenzene.  相似文献   

12.
Soil-borne plant pathogens such as Rhizoctonia solani (Kuhn), Pythium ultimum (Trow) and Sclerotinia trifoliorum (Eriks) can reduce grass and forage legume establishment. The potential for biocontrol of these pathogens by Trichoderma fungi was evaluated. Following dual culture assays, nine Trichoderma isolates (five of Trichoderma atroviride and one each of Trichoderma hamatum, Trichoderma koningiopsis, Trichoderma viride and Trichoderma virens) were chosen for assessment in pot experiments. In the presence of R. solani, perennial ryegrass (Lolium perenne L.) emergence was increased by 60–150% by two isolates of T. atroviride and by 35–212% by the isolate of T. virens, with the increase depending on growing medium and amount of pathogen inoculum. Red clover (Trifolium pratense L.) emergence in the presence of S. trifoliorum was significantly increased by two T. atroviride isolates and the T. hamatum isolate. In the presence of P. ultimum, white clover (Trifolium repens L.) emergence was increased by 25–42% by one isolate of T. atroviride and the T. hamatum isolate. However, for all three pasture species, some Trichoderma isolates reduced seedling emergence. Seedling growth (shoot and root fresh weight/plant) of the three pasture species was significantly increased by one or more T. atroviride isolates. On the basis of these results for both disease reduction and growth promotion, four T. atroviride isolates were selected for field assessment as biocontrol agents of soil-borne pathogens of pasture species.  相似文献   

13.
Adult fungus gnats and moth flies were experimentally demonstrated to function as potential above‐ground vectors for three soilborne plant pathogens: Verticillium dahliae, Fusarium acuminatum and Thielaviopsis basicola. The adult insects externally acquired the conidia of the pathogens after exposure to the cultures as confirmed by scanning electron microscope photography. The intestinal contents and frass deposits of larvae exposed to fungal cultures contained viable fungal propagules. Internally infested larvae developed into internally infested pupae; however, the emerging adults were free of fungal structures. Because of the maintenance of a high level of inoculum on the external body surface and the ability of these adult insects to fly, they can be a significant factor in the dispersal of soilborne fungi in greenhouse agriculture. The rate of dispersal of T. basicola by adult fungus gnats was 1.78 cm2 h?1 per insect and by adult moth flies was 1.17 cm2 h?1 per insect. The area over which the pathogen was dispersed by the adult insects increased with the increase in exposure time. The study demonstrated that adult insects are efficient distributors of soilborne plant pathogenic fungal propagules.  相似文献   

14.
Metabolites produced by the soilborne plant pathogen Rhizoctonia solani (R-23) and Pythium ultimum (PuZ3) grown on cellophane disks placed on potato-dextrose agar (PDA), molasses brewer's yeast agar (MYA) and wheat bran extract agar (BEA) did not significantly affect the rate of growth of isolates of the antagonists Trichoderma viride (TS-I-R3. Tv-101), T. harzianum (Th-57, Th-87), T. hamatum (Tm-23, TRI-4), or Gliocladiun virens (GI-3, GI-21) when these antagonists were grown on the three agars containing pathogen metabolites. However. in some instances. density of antagonist mycelium growing on the agar media as well as the observable production of antagonist conidia on the agar media were reduced. Using four antagonists in liquid cultures of potato-dextrose broth (PDB) containing metabolites of the pathogens grown on bran extract broth, metabolites from R-23 significantly reduced mycelial dry weight of Th-87 and GI-21 but not that of TRI-4 and GI-3. On the contrary. metabolites of PuZ3 increased the mycelial dry weight of all four antagonists, Metabolites of R-23 reduced production of conidia of only TRI-4; metabolites of PuZ3 significantly reduced production of conidia of all four antagonists. Pathogen metabolites did not affect germination of conidia in the system used.  相似文献   

15.
Endophytic isolates of Trichoderma species are being considered as biocontrol agents for diseases of Theobroma cacao (cacao). Gene expression was studied during the interaction between cacao seedlings and four endophytic Trichoderma isolates, T. ovalisporum-DIS 70a, T. hamatum-DIS 219b, T. harzianum-DIS 219f, and Trichoderma sp.-DIS 172ai. Isolates DIS 70a, DIS 219b, and DIS 219f were mycoparasitic on the pathogen Moniliophthora roreri, and DIS 172ai produced metabolites that inhibited growth of M. roreri in culture. ESTs (116) responsive to endophytic colonization of cacao were identified using differential display and their expression analyzed using macroarrays. Nineteen cacao ESTs and 17 Trichoderma ESTs were chosen for real-time quantitative PCR analysis. Seven cacao ESTs were induced during colonization by the Trichoderma isolates. These included putative genes for ornithine decarboxylase (P1), GST-like proteins (P4), zinc finger protein (P13), wound-induced protein (P26), EF-calcium-binding protein (P29), carbohydrate oxidase (P59), and an unknown protein (U4). Two plant ESTs, extensin-like protein (P12) and major intrinsic protein (P31), were repressed due to colonization. The plant gene expression profile was dependent on the Trichoderma isolate colonizing the cacao seedling. The fungal ESTs induced in colonized cacao seedlings also varied with the Trichoderma isolate used. The most highly induced fungal ESTs were putative glucosyl hydrolase family 2 (F3), glucosyl hydrolase family 7 (F7), serine protease (F11), and alcohol oxidase (F19). The pattern of altered gene expression suggests a complex system of genetic cross talk occurs between the cacao tree and Trichoderma isolates during the establishment of the endophytic association.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .  相似文献   

16.
The bronze bug Thaumastocoris peregrinus is an invasive pest, affecting Eucalyptus plantations worldwide. Although its natural enemy Cleruchoides noackae has been tested for the biological control of this pest, other strategies like the use of native entomopathogenic fungi are needed. For this, native virulent fungal isolates should be selected, massively multiplied in an efficient way, and prepared to obtain a stable product. Isolates of native Beauveria bassiana obtained from T. peregrinus and from different collections were screened for their virulence towards this insect and for their amenability to be massively produced in a low-input liquid submerged fermentation and prepared as a dry powder. Three out of six virulent strains were suitable for their massive production in a 2% corn flour suspension, achieving 109 submerged propagules/g of dehydrated preparation. The LC50 achieved by the dry submerged propagules did not differ from the LC50 of fresh aerial conidia. The proposed dual selection of strain and a complex substrate, and the procedures leading to the production of a dry preparation, allowed for high viability and virulence of the fungal spores of three strains.  相似文献   

17.
Extruded granular formulations containing rice flour, gluten, Pyrax, vermiculite, canola oil, and fermentor-produced biomass of isolates of Gliocladium virens (Gl-3, Gl-21 and Gl-32), Trichoderma hamatum (TRI-4 and 31-3), T. harzianum (Th-32 and Th-87) and T. viride (Tv-101) were evaluated for their effect on the reduction of eggplant damping-off caused by Rhizoctonia solani, reduction of pathogen inoculum and proliferation of the isolates in a soil-less mix. Granules with all isolates except 31-3 significantly (P < 0.01) reduced damping-off, and granules with Gl-3, Gl-21, Gl-32, TRI-4 and Th-87 yielded stands comparable to that (90%) of the non-infested control. Granules with isolates Gl-21 and TRI-4 were the most effective in the reduction of saprophytic growth of R. solani, and there was a significant inverse correlation (r 2 = - 0.82) between eggplant stand and saprophytic growth of the pathogen over all treatments. Isolate propagules proliferated to about 107 colony-forming units (CFU) g?1 of soil-less mix after a 6-week incubation, but there was no correlation between the number of CFU and eggplant stand or saprophytic growth reduction of the pathogen. Granules with Gl-21 and TRI-4 amended to pathogen-infested soil-less mix at a rate as low as 0.06% significantly (P < 0.05) reduced damping-off and pathogen saprophytic growth, and a rate of 0.25% of Gl-21 granules resulted in an eggplant stand comparable to that of the non-infested control. There was no significant correlation between the rate of granule amendment and the proliferation of Gl-21 and TRI-4. Granules of Gl-21 and TRI-4 also significantly prevented the spread of R. solani in flats of eggplant seedlings when the biocontrol granules were applied to the soil-less mix 1 day before the pathogen inoculum.  相似文献   

18.
Antagonistic potential of Trichoderma longibrachiatum and T. hamatum against the pathogen Fusarium verticillioides was examined in the laboratory. This was done by pairing each Trichoderma species with the pathogen on 9 cm Petri plates of acidified potato dextrose agar (APDA). Three pairing methods were employed and gradings were assigned to different radial growth suppression of F. verticillioides by each Trichoderma species. Analysis was done using the GLM procedure of the SAS package. Both Trichoderma species significantly inhibited radial growth of F. verticillioides (P = 0.01, R 2 = 0.99) irrespective of pairing method. ‘Inoculating antagonist before pathogen’ supported the best growth inhibition of F. verticillioides by both Trichoderma species. Both Trichoderma species differed significantly (P > 0.0029) in inhibiting radial growth of F. verticillioides. Growth inhibition differed significantly within (P > 0.0059) and among (P > 0.0001) pairing methods. T. longibrachiatum was significantly better than T. hamatum in inhibiting radial growth of F. verticillioides, even at P = 0.01. T. hamatum and T. longibrachiatum could thus be said to show promising antagonistic potential against F. verticillioides with the latter showing better prospects.  相似文献   

19.
Mycoparasitic activities of various isolates of Trichoderma viride, T. harzianum, T. hamatum, T. longibrachiatum, T. koningii, T. pseudokoningii, Gliocladium virens and Laetisaria arvalis were studied against a serious plant pathogen, Botryodiplodia theobromae by scanning electron microscopy (SEM). Macroscopic observations of fungal growth in dual-cultures revealed that most of the isolates made hyphal contact with the pathogen within 2 days after inoculation, leading to the inhibition in pathogen growth. However, T. viride Tv-4, T. hamatum and T. pseudokoningii inhibited pathogen growth before hyphal contact and exhibited an inhibition zone between the colonies of both fungi. SEM investigations demonstrated that in case of hyphal interaction, the firm binding of antagonists (T. viride Tv-1 & Tv-3, T. harzianum Th-1 & Th-2, T. longibrachiatum and L. arvalis) to B. theobromae hyphae established either by coiling around its hyphae, or by penetrating its hyphal cells by forming hooks, haustoria and appressoria-like structures which invariably led to cell disruption. Although T. koningii and G. virens Gv-2 & Gv-3 did not interact physically by way of coiling and penetration, they produced wall lytic enzymes or antifungal substances after coming in contact with B. theobromae which caused wrinkling, bursting and collapsing of pathogen mycelium. It is, therefore, suggested that the outcome of the interaction of antagonist and pathogen was most likely determined by initial hyphal contact that triggered a series of events in pathogen degradation.  相似文献   

20.
Population densities of mycorrhizal fungal propagules in a western Kentucky field highly productive for soybean were measured by bioassay throughout a soybean production season. The primary experimental variables were crop rotation (soybeans in 1985, then 2 years in corn, milo, fescue, or soybean, then soybean in 1988 on all plots when populations of propagules were determined) and soil fumigation with 67% methyl bromide/33% chloropicrin. Of the 20 species in three genera found, Glomus predominated both in terms of number of species and population densities. Most species of Glomus occurred at higher population densities in rotated plots than in continuous soybean plots. In continuous soybean plots, species of Gigaspora made up a much higher proportion of the mycorrhizal fungal community than in rotated crops. Species richness and diversity were lower, and dominance and equitability higher, in nonfumigated continuous soybean plots than in rotated plots early in the season, but the differences were not present at the end of the season. Soil fumigation killed most propagules in the upper 15 cm of soil, but after production of a crop of soybeans, populations of total propagules and most Glomus spp. recovered to prefumigation densities. However, Gigaspora margarita and Gigaspora gigantea did not recover similarly. Fumigation reduced species richness and diversity and increased dominance, but the effects were ameliorated by the end of the season. Colonization of roots was low during vegetative growth but increased rapidly after the onset of soybean reproduction. There was no evidence for mutualism during the early half of the season, perhaps due to high soil P and low dependency of soybean. Fumigation increased soybean yields. A stable mycorrhizal fungal community appeared to become established with continuous soybean production, and both crop rotation and soil fumigation disrupted the community.  相似文献   

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