孙冰冰 杨秀荣 郝永娟 李伟 魏军 田涛

摘 要：为提高生防芽孢杆菌的发酵水平，利用响应面法对生防枯草芽孢杆菌B579进行固相发酵条件优化，以建立一套适合生防芽孢杆菌的固相发酵技术体系。首先通过Plackett-Burman试验筛选出对发酵产量影响显著的重要因素，然后进行最陡爬坡试验逼近最大菌体产量值区域，通过Box-Behnken和响应面试验设计，得到回归模型，并对回归模型进行回归分析和实践验证。结果表明：对发酵产量影响显著的3个重要因素为牛肉膏、玉米粉和可溶性淀粉，当其浓度分别为15.1、10.3 g/kg和9.14 g/kg时，发酵水平达到最高，为1.93×10¹¹cfu/g，比液相发酵水平提高了15倍；经验证该模型准确可靠。

关键词：生防芽孢杆菌；固相发酵；响应面法

中图分类号：S482.2+92文献标识号：A文章编号：1001-4942（2016）05-0062-05

Abstract To enhance the fermentation level of biocontrol Bacillus， the solid-state fermentation conditions of Bacillus subtilis B579 were optimized using response surface methodology to establish the related technology system. Firstly， the Plackett-Burman experiment was applied to screen the importan influential factors， and then the path of steepest ascent was used to approach the biggest region of the cell yield. Moreover， the Box-Benhnken experimental design and response surface methodology were employed to get the regression model， which was proved through regression analysis and practice verification. The results showed that the three factors affecting fermentation production were beef extract， corn meal and soluble starch. The fermentation production reached the highest as 1.93×10¹¹cfu/g， when the content of beef extract， corn meal and soluble starch were 15.1， 10.3 and 9.14 g/kg respectively. Under the optimized fermentation conditions， the solid fermentation production of biocontrol Bacillus increased by 15 times compared to the liquid fermentation. The regression model was verified to be accurate and reliable.

Key words Biocontrol Bacillus； Solid-state fermentation； Response surface methodology

一直以来，化学农药及肥料的使用对于植物病虫害防治和提高农作物产量品质起到重要作用，但大量有害化合物的使用造成了土壤和地下水的严重污染，对生物及人类的生存也造成了严重危害。在过去的几十年中，世界各国都在努力开发可替代传统化学药剂控制植物病虫害的新方法，利用微生物及其代谢产物进行生物防治，被公认为是一种环境友好型的选择。微生物生防菌剂的研究与开发已成为生物防治的一个重要内容。芽孢杆菌是土壤和植物微生态区系的优势生物种群，许多性状优良的菌株表现出对多种植物病原菌的拮抗作用[1～3]。生防芽孢杆菌目前已在水稻、玉米、小麦、大豆、辣椒、棉花、番茄等农作物上显示出较好的病害防治效果[4]。……