钩状木霉的分离鉴定及对辣椒炭疽菌和薏苡黑粉菌的抑菌活性

Isolation,identification of Trichoderma hamatum and determination of its antibacterial activity on Colletotrichum capsici and Ustilago coicis

  • 摘要: 【目的】分离鉴定木霉属有益菌株,探索其对辣椒炭疽菌和薏苡黑粉菌的抑制效果,为木霉属真菌的开发利用及辣椒炭疽病和薏苡黑粉病的生物防治提供理论支持。【方法】采用组织分离法从鱼腥草根际土壤中分离、筛选木霉属有益菌株,通过形态学和内转录间隔区(ITS)测序确定其分类地位,利用两点对峙法、生长速率法、平板对扣法等测试木霉菌菌丝体、小分子代谢产物及挥发性成分对辣椒炭疽菌和薏苡黑粉菌的抑制效果。【结果】采用组织分离法从鱼腥草根际土壤中分离纯化获得单一菌株,记为ZQ202104,结合形态特征和分子生物学鉴定结果,将菌株ZQ202104鉴定为钩状木霉(Trichoderma hamatum)。在对峙培养下,菌株ZQ202104对辣椒炭疽菌的抑制率为59.40%,对薏苡黑粉菌的抑制率为74.01%。生长速率法测定结果显示,菌株ZQ202104菌液加入量为2、4和6 mL时均能很好地抑制辣椒炭疽菌及薏苡黑粉菌的生长,对辣椒炭疽菌的抑制率分别为57.57%、60.60%和65.15%,对薏苡黑粉菌的抑制率分别为71.42%、82.85%和85.71%。菌株ZQ202104无菌发酵液的抑菌活性在发酵9 d时对辣椒炭疽菌的抑制率最高,为35.09%;发酵15 d时对薏苡黑粉菌的抑制率最高,为68.00%。菌株ZQ202104的挥发性成分对辣椒炭疽菌和薏苡黑粉菌的生长均具有抑制作用,抑制率分别为47.29%和82.95%。【结论】从鱼腥草根际土壤中分离到的钩状木霉菌菌株ZQ202104的菌丝体、小分子代谢产物及挥发性成分对辣椒炭疽菌和薏苡黑粉菌均具有抑制作用,特别是对薏苡黑粉菌的生长抑制效果明显,其生防机制可能是竞争作用与抗生作用相结合。菌株ZQ202104具有开发成薏苡黑粉菌抑菌剂的潜力。

     

    Abstract: 【Objective】 The purpose of the study was to isolate and identify beneficial strains of Trichoderma spp., and to explore their inhibitory effects on Colletotrichum capsici and Ustilago coicis, so as to provide a theoretical support for the development and utilization of Trichoderma spp. and the biocontrol of pepper anthracnose and adlay smut disease. 【Method】 The beneficial strains of Trichoderma spp. were isolated and identified from the rhizosphere soil of Houttuyniae herba by tissue isolation method. Its taxonomic status was determined by morphology and ribosomal transcription spacer (ITS) sequencing. The inhibitory effects of Trichoderma spp. mycelium,small molecule metabolites and volatile components on C. capsici and U.coicis were tested by the two-point confrontation method,growth rate method and flat plate buckle method.【Result】 A single strain was isolated and purified from the rhizosphere soil of H. herba by tissue isolation method and recorded as ZQ202104. The results of morphology characteristics and molecular biology identification showed that strain ZQ202104 was T. hamatum. Under the confrontation culture,strain ZQ202104 had inhibitory effect on the growth of C. capsici and U.coicis,and the inhibition rates were 59.40% and 74.01% respectively. The results of growth rate assay showed that strain ZQ202104 could inhibit the growth of C. capsici and U.coicis well when the filtrate was added at the amounts of 2,4 and 6 mL,with the inhibition rates of 57.57%,60.60% and 65.15% against C. capsici and 71.42%,82.85% and 85.71% against U.coicis,respectively. When fermented for 9 d,the bacteriostatic activity of strain ZQ202104 sterile fermentation broth showed the highest inhibition rate on C. capsici(35.09%). When the fermentation was performed for 15 d,the inhibition rate on U.coicis was the highest(68.00%). The volatile components of strain ZQ202104 had inhibitory effects on the growth of C. capsici and U.coicis,and the inhibition rates were 47.29% and 82.95% respectively.【Conclusion】 The mycelium,small molecule metabolites and volatile components of T. hamatum strain ZQ202104 isolated from the soil in the rhizosphere of H. herbahas inhibitory effects on C. capsici and U.coicis, and the growth inhibitory effect is obvious especially for U. coici. The biocontrol mechanism may be a combination of competition and antibacterial effects. Strain ZQ202104 has the potential to be developed into a bacteriostatic agent for U.coicis.

     

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