茶树根腐病菌拮抗真菌的筛选及发酵条件优化

茶树根腐病菌拮抗真菌的筛选及发酵条件优化

DOI:

基金项目:

国家重点研发计划(2022YFD16000803-6)；湖南省重点研发计划(2023NK2013)；湖南省科技创新重大项目(2021NK1020)；国家大学生创新创业训练计划(s202410537115x)

Screening and fermentation condition optimization of an antagonistic fungus against Fusarium cugenangense

Author:

DENG Yulian

Key Laboratory of Tea Science of Ministry of Education, Hunan Agricultural University, Changsha 410128, Hunan, China;College of Horticulture, Hunan Agricultural University, Changsha 410128, Hunan, China
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NIU Li

Key Laboratory of Tea Science of Ministry of Education, Hunan Agricultural University, Changsha 410128, Hunan, China;College of Horticulture, Hunan Agricultural University, Changsha 410128, Hunan, China
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TAN Lin

College of Plant Protection, Hunan Agricultural University, Changsha 410128, Hunan, China
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FAN Jijun

Chenzhou Institute of Agricultural Science, Chenzhou 423001, Hunan, China
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ZHOU Linghong

Chenzhou Institute of Agricultural Science, Chenzhou 423001, Hunan, China
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CUI Wenya

Key Laboratory of Tea Science of Ministry of Education, Hunan Agricultural University, Changsha 410128, Hunan, China;College of Horticulture, Hunan Agricultural University, Changsha 410128, Hunan, China
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HU Qiulong

Key Laboratory of Tea Science of Ministry of Education, Hunan Agricultural University, Changsha 410128, Hunan, China;College of Horticulture, Hunan Agricultural University, Changsha 410128, Hunan, China
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摘要:

【背景】 茶树根腐病是由尖孢镰孢物种复合物(Fusarium oxysporum speciescomplex, FOSC)中一个特异性菌种茶树根腐病原菌镰孢菌(Fusarium cugenangense)引起的一种毁灭性土传病害。【目的】 对茶树根腐病原菌镰孢菌F. cugenangense的拮抗真菌进行分离鉴定，通过优化其培养基和发酵条件以提高其抑菌活性，为后期规模化发酵以及菌剂的开发奠定基础。【方法】 基于形态学及多基因联合建树分析方法对供试菌株进行种属鉴定，测定其抑菌谱，并对其进行发酵条件优化、测定其生长特性及发酵滤液稳定性。【结果】 采用稀释涂布法及平板对峙法筛选出1株茶树根腐病原菌镰孢菌拮抗真菌GF3，其抑菌率达75%；将其初步鉴定为奶灰青霉(Penicillium cremeogriseum)；该菌株对其他10种作物病原菌抑制率达70%以上，具有良好的广谱抑菌性；其难挥发性代谢物和挥发性代谢物对茶树根腐病原菌镰孢菌菌丝的抑制率分别为64%和27%；最适培养基为可溶性淀粉40 g/L、酵母提取物15 g/L、氯化铵6 g/L；最佳发酵条件为pH 6.0、180 r/min、接种量5%，温度28 ℃，时间6 d；于优化后的培养基及培养条件下，其发酵滤液对茶树根腐病原菌镰孢菌的抑菌率达59%，较初始培养基及培养条件的抑菌率增加了33.09%；耐受性试验表明，菌株GF3生长具有良好的耐高温能力、耐盐性、耐碱性能力和紫外耐受性；其发酵滤液在pH 4.0–10.0、温度40–100 ℃条件下均能保持较强的抑菌活性，并且对紫外光照射不敏感，但对K+和Ca2+较敏感。【结论】 确定了菌株GF3的最佳发酵培养基和发酵条件，该菌株及其发酵滤液有较好的拮抗稳定性。

Abstract:

[Background] Tea root rot is a devastating soil-borne disease caused by a specific species Fusarium cugenangense belonging to the Fusarium oxysporum species complex (FOSC). [Objective] To screen out an antagonistic fungus capable of controlling tea root rot and improve its antagonistic activity by optimizing the medium and fermentation conditions, thus laying a foundation for the large-scale fermentation and the development of microbial agents. [Methods] The strain was identified by morphological observation and multi-gene combined analysis, and then the antagonistic spectrum of this strain was determined. Furthermore, the fermentation conditions of the strain were optimized, and its growth characteristics and fermentation filtrate stability were determined. [Results] An antagonistic strain GF3 against F. cugenangense was screened out by dilution coating and plate confrontation methods, with an inhibition rate of 75%. The strain was identified as Penicillium cremeogriseum. Strain GF3 showed the inhibition rates above 70% against other 10 pathogenic fungal species infecting crops, demonstrating a broad antagonistic spectrum. The inhibition rates of refractory volatile metabolites and volatile metabolites of the strain on the hyphae of F. cugenangense were 64% and 27%, respectively. The optimal medium was composed of soluble starch 40 g/L, yeast extract powder 15 g/L, and ammonium chloride 6 g/L, and the optimal fermentation conditions were fermentation with an inoculum amount of 5% at pH 6.0, 180 r/min, and 28 ℃ for 6 days. Under the optimized medium and fermentation conditions, the inhibition rate of strain GF3 against F. cugenangense was 59%, which was 33.09% higher than that under the initial medium and fermentation conditions. Strain GF3 had tolerance to high temperatures, salt, alkali, and ultraviolet. The fermentation filtrate of this strain maintained strong antagonistic activity at pH 4.0–10.0 and 40–100 ℃, and it was not sensitive to ultraviolet light irradiation but sensitive to K+ and Ca2+. [Conclusion] The optimal fermentation medium and fermentation conditions of strain GF3 were determined, and both strain GF3 and its fermentation filtrate had stable antagonistic performance.

邓玉莲,牛丽,谭琳,樊吉君,周玲红,崔文雅,胡秋龙. 茶树根腐病菌拮抗真菌的筛选及发酵条件优化[J]. 微生物学通报, 2025, 52(8): 3753-3771

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