尖镰孢黄瓜专化型枯萎病菌拮抗菌的筛选、鉴定及培养条件优化

doi:10.13560/j.cnki.biotech.bull.1985.2020-0578

摘要/Abstract

摘要： 从蚯蚓粪中筛选出一株能抑制尖镰孢黄瓜专化型枯萎病菌的拮抗菌株,并通过优化培养条件以提高抑菌能力。以尖镰孢黄瓜专化型枯萎病菌为供试病原菌,以蚯蚓粪为供试材料,通过平板稀释法和平板对峙法中筛选到一株能较好抑制尖镰孢黄瓜专化型枯萎病菌的菌株WQ-5,通过形态学观察、生理生化特征和16S rRNA序列分析,初步鉴定其种属。通过单因素和响应面分析法对该菌的培养条件进行优化。通过防效试验验证其防效。菌株WQ-5为解淀粉芽孢杆菌,其最适培养条件为果糖 30 g/L,酵母粉18.9 g/L,硫酸镁3 g/L,甘油1 g/L,磷酸氢二钾1.5 g/L,pH 5.8,装液量38.6%,转速240 r/min,接种量5%和温度30℃。优化后对黄瓜枯萎病菌的抑菌直径和抑菌率较优化前分别提高了24.8%和30.39%。防效试验表明,菌株WQ-5使种子发病率降低了20%,发芽率提高了150%,种子的根长、茎长和活力指数显著提高,植株病情指数下降了48.65%,防治效果为51.67%。菌株WQ-5对黄瓜枯萎病菌有较好防效,具有潜在的研究和应用价值。

关键词: 蚯蚓粪, 拮抗细菌, 尖镰孢黄瓜专化型枯萎病菌, 培养条件, 响应面分析法

Abstract: An antagonistic strain was screened from wormcast and it inhibited the Fusarium oxyporum f. sp. cucumerinum（Foc）,and its inhibitory capability was improved via optimizing culture condition. Using the Foc as experimental pathogen and wormcast as the experimental material,an antagonistic strain named WQ-5 with strong antagonistic activity to Foc was isolated using dilution plate coating and plate confrontation method. The strain WQ-5 was identified by morphological observation,physiological and biochemical characteristics and 16S rRNA sequence analysis. The culture conditions were optimized by single factor and response surface analysis,and its control effect was verified by seed germination test and pot experiment. The strain WQ-5 was identified as Bacillus amyloliquefaciens. The optimized culture conditions,i.e. fructose 30 g/L,peptone 18.9 g/L,magnesium sulfate 3 g/L,glycerol 1 g/L,dipotassium hydrogen phosphate 1.5 g/L,pH 5.8,liquid-loading rate 38.6%,rotating speed 240 r/min,inoculation rate 5%,temperature 30℃ and cultural time 24 h,increased inhibition zone diameter by 24.8% and the inhibition rate of pathogens by 30.39%,as compared to the non-optimized conditions. The control effect experiment showed that the strain WQ-5 reduced the disease incidence of seeds by 20%,increased germination rate by 150%,significantly enhanced the root length,stem length and vigor index of seeds,decreased the disease index of plants by 48.65%,and decreased the disease index of plants by 51.67%. In conclusion,the strain WQ-5 demonstrates promising control effect on Fusarium wilt caused by Foc and presents potential research and application value.

Key words: wormcast, antagonistic bacterium, Fusarium oxyporum f. sp. cucumerinum, culture conditions, response surface analysis

张美君, 吴庆, 尹翠, 王妮, 马晓庆, 马晓霞, 曹云娥. 尖镰孢黄瓜专化型枯萎病菌拮抗菌的筛选、鉴定及培养条件优化[J]. 生物技术通报, 2020, 36(9): 125-136.

ZHANG Mei-jun, WU Qing, YIN Cui, WANG Ni, MA Xiao-qing, MA Xiao-xia, CAO Yun-e. Screening and Identification of an Antagonistic Strain Against Fusarium oxyporum f. sp. cucumerinum and Optimization of Culture Conditions[J]. Biotechnology Bulletin, 2020, 36(9): 125-136.

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