倪敏姿, 陈涛, 崔峰, 陈杰. 棘孢木霉TCS007菌株固体发酵产孢条件优化及其抑菌活性[J]. 农药学学报. DOI: 10.16801/j.issn.1008-7303.2024.0051
    引用本文: 倪敏姿, 陈涛, 崔峰, 陈杰. 棘孢木霉TCS007菌株固体发酵产孢条件优化及其抑菌活性[J]. 农药学学报. DOI: 10.16801/j.issn.1008-7303.2024.0051
    NI Minzi, CHEN Tao, CUI Feng, CHEN Jie. Optimization of solid-state fermentation for spore production of Trichoderma asperellum TCS007 strain and its antifungal activity[J]. Chinese Journal of Pesticide Science. DOI: 10.16801/j.issn.1008-7303.2024.0051
    Citation: NI Minzi, CHEN Tao, CUI Feng, CHEN Jie. Optimization of solid-state fermentation for spore production of Trichoderma asperellum TCS007 strain and its antifungal activity[J]. Chinese Journal of Pesticide Science. DOI: 10.16801/j.issn.1008-7303.2024.0051

    棘孢木霉TCS007菌株固体发酵产孢条件优化及其抑菌活性

    Optimization of solid-state fermentation for spore production of Trichoderma asperellum TCS007 strain and its antifungal activity

    • 摘要: 棘孢木霉 (Trichoderma asperellum) TCS007菌株分离自海洋极端环境并进行了驯化培养和诱变稳定,该菌株及其代谢产物对植物病原真菌具有广谱的抑菌活性,且能诱导植物促生、抗逆等,具有较好的开发前景。为探索TCS007菌株固体发酵产孢量的最佳发酵条件,明确其代谢产物的抑菌活性,本研究通过单因素试验,结合响应曲面法,对TCS007菌株固体发酵培养基配方和发酵培养条件进行优化;采用菌丝生长速率法测定其发酵滤液和粗提物对6种植物病原真菌的抑菌活性。结果表明:影响TCS007菌株固体发酵产孢量的3个主要因素为培养时间、加水量和光照周期。优化后的发酵培养基配方为麦麸和黄豆粉按质量比5 : 1混合,并添加1%硫酸锰、1%硫酸镁;优化后的发酵条件为培养时间15 d、加水量39.7 mL、光照时间21 h、培养温度为28 ℃、发酵接种量2 mL (1 × 107个/mL)、初始pH值5.0。在此条件下,按优化后的发酵培养基配方,TCS007菌株的孢子产量为7.14 × 109个/g,与响应面法预测的孢子产量 (6.85 × 109个/g) 拟合,比基础发酵条件孢子产量提高了6倍。抑菌活性测定结果表明,TCS007菌株固体发酵滤液(稀释10倍)和粗提物 (10 mg/L) 对油菜菌核病菌Sclerotinia sclerotiorum均具有较好的抑制效果,抑制率分别为92.4%和95.8%。该结果可为后续产品发酵工艺研究及其木霉菌剂的研制提供理论依据。

       

      Abstract: Trichoderma asperellum TCS007 strain was isolated from marine extreme environments and was domesticated and mutagenic stabilized. It is known that this strain and its metabolites have broad-spectrum inhibitory activity against plant pathogenic fungi and have the capabilities of inducing plant growth and resistance, exhibiting a great development prospect. In order to explore the optimal fermentation conditions for the spore production of TCS007 strain in solid-state fermentation, and define the antifungal activity of the metabolites of TCS007 strain, the formulation of TCS007 strain solid fermentation medium and fermentation culture conditions were optimized by a one-way test combined with the response surface method, and then the antifungal activities of the solid-state fermentation filtrate and crude extract against six plant pathogenic fungi were determined by the mycelial growth rate method. The experimental results showed that the three main factors affecting the spores yield of solid fermentation of TCS007 strain were incubation time, water addition and light cycle time. The optimized fermentation substrate was wheat bran + soybean meal, with a mass ratio of 5 : 1, supplied with 1% MnSO4 and 1% MgSO4; the optimized fermentation conditions were 15 d of incubation time, 39.7 mL of water addition, 21 h of light cycle time, 28 ℃ of culture temperature, 2 mL of inoculum (1 × 107 spores/mL), and the initial pH value 5.0. Under these fermentation conditions, the spore yield of TCS007 strain was 7.14 × 109 spores/g, which was fitted to the spore yield predicted by the response surface methodology (6.85 × 109 spores/g), and a 6-fold increase in spore yield over the initial fermentation condition. The antifungal assays further showed that both TCS007 strain solid-state fermentation filtrate (10-fold dilution) and crude extract (10 mg/L) showed good inhibitory effect against Sclerotinia sclerotiorum, with inhibition rates of 92.4% and 95.8%, respectively. These findings will provide the theoretical basis for the study of the fermentation process of the subsequent products and the development of TCS007 strain fungicide.

       

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