A new high-performance liquid chromatography-tandem mass spectrometry method based on dispersive solid phase extraction for the determination of the mycotoxin fusarin C in corn ears and processed corn samples

Fusarin C is a mycotoxin that is produced by a variety of Fusarium species and is therefore a possible contaminant in food and feed. For this reason, a reliable high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of fusarin C in food and feed samples was developed based on dispersive solid phase extraction (DSPE). This method has a limit of detection (LOD) of 2 μg/kg, a limit of quantitation (LOQ) of 7 μg/kg, and a recovery rate of 80%. Fifty different corn samples were analyzed, and fusarin C was detected in 40 of them. The fusarin C level varied in kernels of corn ears from not detectable up to 83 mg/kg and in food samples from not detectable up to 28 μg/kg. The co-occurrence of further structural analogues of fusarin C was confirmed by high-performance liquid chromatography Fourier transformation mass spectrometry (HPLC-FTMS). In addition, the stability of fusarin C under storage conditions was evaluated.     

红三叶根际促生菌中具生防效果菌株筛选、鉴定及特性研究

【目的】 从岷山红三叶根际分离的109株促生菌,鉴定、筛选了具有生防能力的菌种资源并研究其生防特性,为生产应用提供服务。 【方法】 采用平板对峙法筛选优良生防菌株,对其产铁载体能力进行定性、定量测定和16S rRNA基因序列分析鉴定。 【结果】 平板对峙试验显示,109株促生菌中有8株促生菌对3种病原真菌 (立枯丝核菌、黄瓜枯萎菌、西瓜尖镰孢菌) 具有拮抗效果,分别是菌株MHS3、MHS7、MHS9、MHS27、MHS31、MHS38、MHS39和MHS48。其中,菌株MHS27、MHS31发酵液对立枯丝核菌抑菌率分别为40.1%、47.1%;菌株MHS7和MHS48发酵液对黄瓜枯萎菌抑菌率分别为26.1%、22.8%;菌株MHS27和MHS31发酵液对西瓜尖镰孢菌抑制率分别为34.0%、30.5%;产嗜铁素是生防菌 (MHS7、MHS27、MHS31、MHS48) 拮抗3种病原真菌的主要途径之一。经16S rRNA基因序列分析初步鉴定,菌株MHS7为特基拉芽孢杆菌 (Bacillus tequilensis),菌株MHS27为溶蛋白芽孢杆菌 (Bacillus proteolyticus),菌株MHS31为Pseudomonas paralactis,菌株MHS48为克雷伯氏菌 (Klebsiella oxytoca)。 【结论】 从岷山红三叶根际分离筛选出4株具有优良生防作用的PGPR菌株,为今后生物菌肥应用推广提供了菌种资源支持。      

Metabolites inhibiting germination of Orobanche ramosa seeds produced by Myrothecium verrucaria and Fusarium compactum

Myrothecium verrucaria and Fusarium compactum were isolated from diseased Orobanche ramosa plants collected in southern Italy to find potential biocontrol agents of this parasitic weed. Both fungi grown in liquid culture produced metabolites that inhibited the germination of O. ramosa seeds at 1-10 muM. Eight metabolites were isolated from M. verrucaria culture extracts. The main metabolite was identified as verrucarin E, a disubstituted pyrrole not belonging to the trichothecene group. Seven compounds were identified by spectroscopic methods as macrocyclic trichothecenes, namely, verrucarins A, B, M, and L acetate, roridin A, isotrichoverrin B, and trichoverrol B. The main metabolite produced by F. compactum was neosoloaniol monoacetate, a trichothecene. All the trichothecenes proved to be potent inhibitors of O. ramosa seed germination and possess strong zootoxic activity when assayed on Artemia salina brine shrimps. Verrucarin E is inactive on both seed germination and zootoxic assay.     

Structure elucidation of new fusarins revealing insights in the rearrangement mechanisms of the Fusarium mycotoxin fusarin C

Fusarin C is a Fusarium mycotoxin that rearranges under reversed phase chromatographic conditions. In this study, the rearrangement of fusarin C was examined in detail, and the formation of fusarins under different conditions was optimized. All relevant fusarins including (10Z)-, (8Z)-, and (6Z)-fusarin C were isolated and identified by NMR. To confirm the involvement of the 2-pyrrolidone ring in the rearrangement of fusarin C, 15-methoxy-fusarin C was synthesized. For the first time, the structure of open-chain fusarin C was elucidated, and on the basis of these data, the rearrangement product of fusarin C was identified as epi-fusarin C. The results were confirmed by detailed NMR measurements and density functional theory calculations. Furthermore, a new fusarin C like metabolite, which was named dihydrofusarin C, was detected by analysis of the crude extract of fusarin C with high-performance liquid chromatography coupled to UV and Fourier transform mass spectrometry.     

Production of bacillomycin- and macrolactin-type antibiotics by Bacillus amyloliquefaciens NJN-6 for suppressing soilborne plant pathogens

Bacillus amyloliquefaciens strains have been used as biocontrol agents for the suppression of several soilborne plant pathogens. A clearer understanding of the antagonistic mechanisms of action of these bacteria will facilitate their use in the control of plant diseases. Antagonistic substances were isolated from the fermentation broth of B. amyloliquefaciens strain NJN-6 cultures. These compounds were preconcentrated using an XAD-16 column and were purified using reversed-phase high-performance liquid chromatography (RP-HPLC). Fractions were collected from the column and were analyzed, and two homologues of bacillomycin D [molecular weights of 1030 Da (C14) and 1044 Da (C15)] and three homologues of members of the macrolactin family, macrolactin A, 7-O-malonyl macrolactin A, and 7-O-succinyl macrolactin A (molecular weights of 402, 487, and 502 Da, respectively) were identified using HPLC/electrospray ionization mass spectrometry (ESI-MS) analysis. An antagonistic assay showed that bacillomycin D and macrolactin exhibited significant antagonistic effects against Fusarium oxysporum and Ralstonia solanacearum , respectively. A reliable method for the isolation and purification of bacillomycin D and macrolactin from bacterial broth cultures was developed. These data will help elucidate the mechanisms that B. amyloliquefaciens NJN-6 uses for the biocontrol of soilborne plant pathogens.     

Antifungal properties of wheat histones (H1-H4) and purified wheat histone H1

Wheat ( Triticum spp.) histones H1, H2, H3, and H4 were extracted, and H1 was further purified. The effect of these histones on specific fungi that may or may not be pathogenic to wheat was determined. These fungi included Aspergillus flavus , Aspergillus fumigatus , Aspergillus niger , Fusarium oxysporum , Fusarium verticillioides , Fusarium solani , Fusarium graminearum , Penicillium digitatum , Penicillium italicum , and Greeneria uvicola . Non-germinated and germinating conidia of these fungi were bioassayed separately. The non-germinated and germinating conidia of all Fusarium species were highly susceptible to the mixture (H1-H4) as well as pure H1, with viability losses of 99-100% found to be significant (p < 0.001) at ≤10 μM or less for the histone mixture and pure H1. F. graminearum was the most sensitive to histone activity. The histones were inactive against all of the non-germinated Penicillium spp. conidia. However, they significantly reduced the viability of the germinating conidia of the Penicillium spp. conidia, with 95% loss at 2.5 μM. Non-germinated and germinating conidia viability of the Aspergillus spp. and G. uvicola were unaffected when exposed to histones up to 10 μM. Results indicate that Fusarium spp. pathogenic to wheat are susceptible to wheat histones, indicating that these proteins may be a resistance mechanism in wheat against fungal infection.     

Disease control effect of strevertenes produced by Streptomyces psammoticus against tomato fusarium wilt

During screening of microorganisms producing antifungal metabolites, Streptomyces psammoticus strain KP1404 was isolated. The culture extract of this strain showed potent disease control efficacy against Fusarium wilt on tomato plants. The antifungal metabolites ST-1 and ST-2 were isolated from the culture extract using a variety of chromatographic procedures. On the basis of MS and NMR spectrometric analysis, the structures of the antifungal active compounds ST-1 and ST-2 were determined to be the polyene antibiotics strevertene A and strevertene B, respectively. In vitro, strevertenes A and B showed inhibitory effects against the mycelial growth of Alternaria mali , Aspergillus oryzae , Cylindrocarpon destructans , Colletotrichum orbiculare , Fusarium oxysporum f.sp. lycopersici, and Sclerotinia sclerotiorum , even at concentrations of 4-16 μg/mL. Fusarium wilt development on tomato plants was strongly retarded by treatment with 1 μg/mL of these strevertenes. The disease control efficacies of strevertenes on Fusarium wilt were as remarkable as that of benomyl.     

温室及田间条件下解淀粉芽孢杆菌菌株W19对香蕉生长、产量的促进和对香蕉镰刀菌萎蔫病的抑制研究

Plant growth-promoting rhizobacteria (PGPR) are considered to be the most promising agents for cash crop production via increasing crop yields and decreasing disease occurrence. The Bacillus amyloliquefaciens strain W19 can produce secondary metabolites (iturin and bacillomycin D) effectively against Fusarium oxysporum f. sp. cubense (FOC). In this study, the ability of a bio-organic fertilizer (BIO) containing W19 strain to promote plant growth and suppress the Fusarium wilt of banana was evaluated in both pot and field experiments. The results showed that application of BIO significantly promoted the growth and fruit yield of banana while suppressing the banana Fusarium wilt disease. To further determine the beneficial mechanisms of the strain, the colonization of green fluorescent protein-tagged strain W19 on banana roots was observed using confocal laser scanning microscopy and scanning electron microscopy. The effect of banana root exudates on the formation of biofilm of strain W19 indicated that the banana root exudates may enhance colonization. In addition, the strain W19 was able to produce indole-3-acetic acid (IAA), a plant growth-promoting hormone. The results of these experiments revealed that the application of strain W19-enriched BIO improved the banana root colonization of strain W19 and growth of banana and suppressed the Fusarium wilt. The PGPR strain W19 can be a useful biocontrol agent for the production of banana under field conditions.     

Cyclopenta[b]benzofurans from Aglaia species with pronounced antifungal activity against rice blast fungus (Pyricularia grisea)

Eight flavaglines, six cyclopenta[b]benzofurans, a cyclopenta[bc]benzopyran, and a benzo[b]oxepine, together with an aromatic butyrolactone were isolated from Aglaia odorata, A. elaeagnoidea, and A. edulis (Meliaceae) and tested against the three plant pathogens Pyricularia grisea, Fusarium avenaceum, and Alternaria citri for antifungal properties. Using the microdilution technique linked with digital image analysis of germ tubes, the benzofurans displayed strong activity, whereas the benzopyran, benzoxepine, and butyrolactone were inactive at the highest concentration tested. P. grisea, responsible for rice blast disease, was the most susceptible fungus against all benzofurans, with rocaglaol as the most active derivative. Based on EC(50), EC(90), and MIC values, the antifungal activity of rocaglaol was clearly higher than of the reference compounds, blasticidin S and Benlate.     

Volatiles from Fusarium verticillioides (Sacc.) nirenb. and their attractiveness to nitidulid beetles

It is known that sap beetles (Coleoptera: Nitidulidae) can vector the fungus Fusarium verticillioides (Sacc.) Nirenb. (= F. moniliforme Sheldon), which causes an important ear-rot disease in corn and also produces fumonisin mycotoxins. The volatiles produced by this fungus were studied to establish whether they could attract sap beetles. Such an association would suggest more than just an incidental role in transmission of the fungus by the beetles. F. verticillioides consistently produces a blend of five alcohols (ethanol, 1-propanol, 2-methyl-1-propanol, 3-methyl-1-butanol, and 2-methyl-1-butanol), acetaldehyde, and ethyl acetate. Ethanol is the most abundant alcohol. The fungus also produces four phenolic compounds (the most abundant of which is ethylguaiacol), a series of presently unidentified sesquiterpene hydrocarbons, and an unidentified compound that is probably a 10-carbon ketone. Solid-phase microextraction was the key technique used in volatile analysis. The volatile profiles change over time and differ somewhat among fungal strains: The alcohols, aldehyde, and ester always appeared first and were present for each strain. Production of the phenolics lagged by several days, and in some strains these compounds were barely detectable. Volatile production eventually diminished in all strains. All strains were attractive to the sap beetle, Carpophilus humeralis (F.), in wind-tunnel bioassays. Attraction was correlated primarily to the presence of the alcohols, acetaldehyde, and ethyl acetate, rather than to the phenolics. To verify that the identified culture volatiles were responsible for beetle attraction, cultures were quantitatively simulated with synthetic chemicals, and the cultures and corresponding synthetic mixtures were then compared by bioassay. The comparisons were favorable. Volatile emission patterns from cultures were fairly robust with respect to inoculum level or incubation temperature, but some manipulation was possible. For example, after freeze-drying and rehydrating (a rapid simulation of winter/spring conditions), F. verticillioides produced ethyl acetate and other esters at unusually high levels. The fungus produced attractive volatiles following ear inoculation of milk-stage field corn as well as on sterile, mature kernels in the laboratory.     

Isolation and structural elucidation of eight new related analogues of the mycotoxin (-)-botryodiplodin from Penicillium coalescens

Bioassay-guided fractionation of the organic extract derived from the terrestrial fungus Penicillium coalescens led to the isolation of the known mycotoxin (-)-botryodiplodin (1) and eight new structurally related analogues (2-9). The structures of the novel compounds were determined by MS and NMR studies, including 1D and 2D NMR. A likely biogenetic pathway from the aldehydic open form of 1 (C7 unit, U1) is proposed for these metabolites. Among all the isolated metabolites, only (-)-1 showed antifungal, antibacterial, and insecticidal activity. This latter activity appears to be a new property attributed to (-)-1.     

Biocontrol efficiency of Fusarium wilt diseases by a root-colonizing fungus Penicillium sp.

Soil-inhabiting fungal pathogen Fusarium oxysporum often causes severe yield losses in many crops. We investigated the effect of a plant growth-promoting fungus, Penicillium sp. EU0013 on Fusarium wilt disease. In dual culture experiments, EU0013 inhibited the growth of Fusarium wilt pathogens by producing an inhibition zone. In experiments using sterile potting medium under controlled conditions, EU0013 significantly reduced the severity of Fusarium wilt on tomato (Solanum lycopersicum L.) and cabbage (Brassica oleracea L. var. capitata). In non-sterile soil, benomyl-resistant mutants of EU0013 were selected by exposing the conidial solution of EU0013 to ultraviolet light. The selected mutant EU0013_90S isolate did not show any distinct differences from EU0013 in colony characteristics, growth rate or antifungal activity against Fusarium wilt pathogens in dual culture. The effect of EU0013_90S on tomato wilt was studied under greenhouse conditions using non-sterile soil. Two-weeks old tomato seedlings were dipped in four different concentrations of EU0013_90S conidial suspension (1?×?10³, 1?×?10⁴, 1?×?10⁵, and 1?×?10⁶ conidia mL^–1). Seedlings were then planted in soil inoculated with either F. oxysporum f. sp. lycopersici race 1 CU1 or race 2 JCM 12575 (1?×?10⁶ bud-cells g^–1). We found the greatest disease suppression occurred when seedlings were dipped in the highest concentration of EU0013_90S conidia. This same inoculum concentration of EU0013_90S also resulted in the highest disease reduction in soil infested with JCM 12575. Higher root colonization with EU0013_90S showed a significant reduction in Fusarium wilt disease, suggesting that colonization by Penicillium sp. EU0013_90S is important for efficient biocontrol of these diseases.     

Degradation of Sevin by soil microorganisms

Microorganisms capable of transforming the pesticide 1-naphthyl N-methyl-carbamate (Sevin) were isolated from soil. Three isolates were able to accelerate the hydrolysis of Sevin to 1-naphthol. Several unidentified intermediates were separated by thin-layer chromatography and also by following the decomposition of Sevin-methyl-¹⁴C. Since 1-naphthol is a biological as well as a chemical decomposition product of Sevin, its transformation by the isolated microbes was also studied. A fungus, identified as Fusarium solani, altered 1-naphthol rapidly. Whereas one strain of bacterium degraded the hydrolysis product gradually, another strain accumulated it under certain conditions. Mixed cultures of the investigated microbes were more effective in transforming Sevin than pure cultures, and this phenomenon was also observed with 1-naphthol as substrate with the exception of one bacterial strain.     

Fumonisin production and bioavailability to maize seedlings grown from seeds inoculated with Fusarium verticillioides and grown in natural soils

The fungus Fusarium verticillioides infects maize and produces fumonisins. The purpose of this study was to determine the ability of F. verticillioides to produce fumonisins in synthetic and natural soils and their biological availability to maize roots. Maize seeds were inoculated with a pathogenic strain of F. verticillioides (MRC826) and planted in synthetic and three different natural soils. There were statistically significant reductions in stalk weight and root mass and increased leaf lesions in the MRC826-treated seedlings in all soil types. Fumonisins were detected in all of the soils of seedlings grown from MRC826-inoculated seeds. The fumonisin produced in the soils was biologically available to seedlings as demonstrated by the statistically significant elevation of free sphingoid bases and sphingoid base 1-phosphates in their roots. These results indicate that F. verticillioides produced fumonisins in the autoclaved synthetic and natural soils and that the fumonisin produced is biologically available on the basis of evidence of inhibition of ceramide synthase.     

Production of fumonisin B and C analogues by several fusarium species

Six strains of Fusarium verticillioides, two of F. oxysporum, one strain of F. proliferatum, and a strain of an unidentified species were cultured on maize patties and rice and evaluated for their ability to simultaneously produce fumonisin B (FB) and C (FC) series analogues. Fumonisins were quantified by LC-MS-MS using positive ion electrospray ionization. FC1 provided characteristic fragment ions at m/z 690, 672, 654, 532, 514, and 338 corresponding to sequential loss of H2O and tricarboxylic acid moieties from the alkyl backbone, while FC3 and FC4 provided equivalent product ions 16 and 32 amu lower than the corresponding FC1 fragments, respectively. All isolates cultured on maize produced FC4. All isolates except for that of F. proliferatum also produced FC1, and three of the six strains of F. verticillioides produced FC3. All isolates except those of F. oxysporum produced detectable amounts of FB1, FB2, and FB3. Isolates that produced fumonisin B analogues produced at least 10 fold more of the B series analogues than they did of the C series analogues. The results confirm that at least some strains of F. oxysporum produce FC, but not FB, fumonisin analogues and also suggest that the genetics and physiological regulation of fumonisin production may be more complicated than previously envisaged since some strains of F. verticillioides and F. proliferatum as well as the strain of the unidentified species can simultaneously produce both FB and FC analogues.     

Conversion of the mycotoxin patulin to the less toxic desoxypatulinic acid by the biocontrol yeast Rhodosporidium kratochvilovae strain LS11

The infection of stored apples by the fungus Penicillium expansum causes the contamination of fruits and fruit-derived products with the mycotoxin patulin, which is a major issue in food safety. Fungal attack can be prevented by beneficial microorganisms, so-called biocontrol agents. Previous time-course thin layer chromatography analyses showed that the aerobic incubation of patulin with the biocontrol yeast Rhodosporidium kratochvilovae strain LS11 leads to the disappearance of the mycotoxin spot and the parallel emergence of two new spots, one of which disappears over time. In this work, we analyzed the biodegradation of patulin effected by LS11 through HPLC. The more stable of the two compounds was purified and characterized by nuclear magnetic resonance as desoxypatulinic acid, whose formation was also quantitated in patulin degradation experiments. After R. kratochvilovae LS11 had been incubated in the presence of (13)C-labeled patulin, label was traced to desoxypatulinic acid, thus proving that this compound derives from the metabolization of patulin by the yeast. Desoxypatulinic acid was much less toxic than patulin to human lymphocytes and, in contrast to patulin, did not react in vitro with the thiol-bearing tripeptide glutathione. The lower toxicity of desoxypatulinic acid is proposed to be a consequence of the hydrolysis of the lactone ring and the loss of functional groups that react with thiol groups. The formation of desoxypatulinic acid from patulin represents a novel biodegradation pathway that is also a detoxification process.     

Structure-activity relationships of derivatives of fusapyrone, an antifungal metabolite of Fusarium semitectum

Fusapyrone (1) and deoxyfusapyrone (2) are two 3-substituted-4-hydroxy-6-alkyl-2-pyrones isolated from Fusarium semitectum that have considerable antifungal activity against molds. Because of their low zootoxicity and selective action they are potentially utilizable along with biocontrol yeasts for control of postharvest crop diseases. Seven derivatives of 1 (3 and 5-10) and one derivative of 2 (4) were obtained by chemical modifications of the glycosyl residue, the 2-pyrone ring, the aliphatic chain, or a combination thereof, and a structure-activity correlation study was carried out with regard to their zootoxicity and antifungal activity. Derivatives 7-10, as well as 1, were slightly zootoxic in Artemia salina (brine shrimp) bioassays, whereas pentaacetylation of 1 into 3, 5, and 6 resulted in a strong increase in toxicity. Compound 4, the tetraacetyl derivative of 2, was as toxic as 2. Because the structural changes of 1 that resulted in an increase of biological activity in A. salina bioassay were those that affected mainly the water solubility of the molecule, it appears that toxicity is related to hydrophobicity. Compounds 1 and 2 showed strong antifungal activity toward Botrytis cinerea, Aspergillus parasiticus, and Penicilliun brevi-compactum (minimum inhibitory concentration at 24 h = 0.78-6.25 microg/mL). Among derivatives 3-10, only compounds 7, 9, and 10 retained some activity, limited to B. cinerea and at high concentration (25-50 microg/mL). None of the compounds 1-10 inhibited the growth of the biocontrol yeasts Pichia guilliermondii and Rhodotorula glutinis at the highest concentration tested (50 microg/mL).     

NTG诱导及核辐射改良稗草生防潜力菌研究

用亚硝基胍 (nitrosoguanidine ,NTG)诱变稗草生防潜力菌禾长蠕孢稗草专化型菌株(HGE) ,获得诱变菌株I2 62 ,其孢子产量比原始菌株 (HGE)提高 5 2 6%。以I2 62 为出发菌株 ,用60 Coγ射线 65 0Gy辐照 5 7min ,有 7 75 %的菌株产孢量高于出发菌株高 ,其中辐照菌株FⅡ12 1、FⅡ116和FⅡ14 0 的产孢量分别比I2 62 提高 5 4 4%、5 1 5 %和 41 7%。化学和物理技术相结合处理稗草生防潜力菌 ,获得的突变菌株产孢量比原始菌株提高 1倍以上。但高产孢突变菌株对稗草的致病性、防效与原始菌株相当。     

Transformation of Eutypa dieback and esca disease pathogen toxins by antagonistic fungal strains reveals a second detoxification pathway not present in Vitis vinifera

Eutypine, 4-hydroxybenzaldehyde, and 3-phenyllactic acid are some of the phytotoxins produced by the pathogens causing Eutypa dieback and esca disease, two trunk diseases of grapevine (Vitis vinifera). Known biocontrol agents such as Fusarium lateritium and Trichoderma sp. were screened for their ability to consume these toxins. Transformation time courses were performed, and an high-performance liquid chromatography-based method was developed to analyze toxin metabolism and to identify and quantify the converted products. The results show that the aldehyde function of eutypine was reduced to eutypinol, as by V. vinifera cv. Merlot, the cultivar tolerant to Eutypa dieback. We revealed a supplementary detoxification pathway, not known in Merlot, where the aldehyde function was oxidized to eutypinic acid. Moreover, some strains tested could further metabolize the transformation products. Every strain tested could transform 4-hydroxybenzaldehyde to the corresponding alcohol and acid, and these intermediates disappeared totally at the end of the time courses. When biological assays on cells of V. vinifera cv. Chasselas were carried out, the transformation products exhibited a lower toxicity than the toxins. The possibility of selecting new biocontrol agents against trunk diseases of grapevine based on microbial detoxification is discussed.     

钩状木霉 ACCC31649的GFP标记及其对辣椒定殖和促生作用

【目的】旨在建立农杆菌介导的绿色荧光蛋白 (GFP) 基因转化钩状木霉 (Trichoderma hamatum) ACCC31649的技术方法,筛选遗传稳定的GFP标记转化子,并研究该菌株在辣椒植株中定殖和对辣椒的促生作用,为进一步阐明木霉在辣椒根际定殖及其与辣椒病原菌在辣椒根际和植株中的定殖、互作和生防作用奠定基础。【方法】通过根癌农杆菌介导的遗传转化方法筛选遗传稳定的GFP标记转化子,通过灌根接种方法和组织切片的水玻片荧光观察研究了钩状木霉在辣椒植株中定殖过程和对辣椒的促生作用。【结果】获得了遗传稳定GFP标记的钩状木霉转化子。荧光显微观察表明,辣椒根、茎、叶组织中都检测到GFP标记菌株的定殖。标记菌株首先在根部定殖,然后通过根部维管束逐步定殖到茎和叶片组织中。野生型菌株和GFP标记菌株灌根接种4叶期辣椒幼苗,30天后,GFP标记菌株与水处理对照相比,辣椒的株高增长13.5%,根长增长16.2%,鲜重和干重分别增加了43.8%和45.3%,而且野生型菌株与GFP标记菌株对辣椒的促生作用没有显著差异。【结论】钩状木霉能够在辣椒植株根、茎和叶组织中定殖,并且对辣椒具有显著的促生作用。同时,GFP标记的钩状木霉将在进一步阐明该菌株在辣椒根际定殖及其对病原菌拮抗和互作研究中发挥重要作用。