烟草赤星病拮抗芽胞杆菌的筛选及培养基优化

罗楚翔; 彭建; 杨欢; 陈建刚; 王昌军; 李进平; 施河丽; 祁高富; 陈守文

doi:10.13496/j.issn.1007-5119.2014.03.011

烟草赤星病拮抗芽胞杆菌的筛选及培养基优化

Screening and Medium Optimization of Antagonistic Bacillus Against Tobacco Brown Spot Disease

摘要

摘要: 以赤星病原菌Alternaria longipes为筛选指示菌，采用平板共培养初筛和发酵上清滤液复筛的方法，筛选了烟草赤星病拮抗芽胞杆菌3株。其中菌株K₁₁的菌体径向生长抑制率（PIRG）为63.64%，发酵上清滤液的PIRG值达67.44%。盆栽试验证明，该菌对烟草赤星病具有良好的拮抗能力。经鉴定菌株K₁₁为枯草芽胞杆菌（Bacillus subtilis）。由单因素实验和正交实验优化得到了K₁₁菌株生物量较高的摇瓶发酵培养基配方：葡萄糖20.0 g/L，豆粕30.0 g/L，K₂HPO₄·3H₂O 1.0 g/L，MgSO₄·7H₂O 0.1 g/L。优化后生防菌株K₁₁的生物量比对照提高6.5倍，生物量高达1.28×10¹⁰CFU/mL，芽胞形成率在95%以上。

Abstract: Three strains were selected as the Bacillus agtagonists against tobacco brown spot disease from 93 strains stored in our lab, the strain K₁₁ showed the highest percentage inhibition of radial growth (PIRG). The PIRG was 63.64% by the dual culture while 67.44% in culture filtrate tests. According to the physiological and biochemical characteristics, the strain K₁₁was identified as Bacillus subtilis. Based on the single factor experiments and orthogonal experiments for K₁₁, an optimum fermentation medium for biomass production in flasks composed of glucose 20.0 g/L, soybean meal 30.0 g/L, K₂HPO₄·3H₂O 1.0 g/L, and MgSO₄·7H₂O 0.1 g/L. Under the optimum fermentation conditions, the highest biomass yield was 1.28×10¹⁰ CFU/mL, which was 6.5 times of the initial KMB medium, and the sporulation rate reached as much as 95%.

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