响应面法优化绿色木霉H06产孢发酵培养基.doc

响应面法优化绿色木霉H06产孢发酵培养基 梁昌聪，刘磊，张建华，王伟伟，黄俊生﹡ (中国热带农业科学院环境与植物保护研究所/农业部热带作物有害生物综合治理重点实验室/海南省热带农业有害生物检测与控制重点实验室，海南 海口 571101) 摘 要:采用响应面法对绿色木霉H06菌株产孢发酵培养基进行了优化。首先利用Plackett－Burman实验设计筛选出影响产孢的 3个主要因素: 玉米粉、大豆粕和 KH2PO4。在此基础上运用最陡爬坡路径法逼近最大响应值区域，最后利用响应面分析法确定主要因子之间的交互作用及最佳条件。结果表明，蔗糖10g/L，玉米粉12.75 g/L，NH4NO32 g/L，大豆粕4.65 g/L，MgSO4·7H2O 1 g/L，KH2PO43.32 g/L，H06最大理论孢子含量为3.29×109个/mL。经3次平行实验验证，实际平均孢子含量与预测孢子含量相近，比之前的孢子含量提高了197%。 关键词：响应面法，绿色木霉，产孢，优化 Optimization of medium components for sporulation quantity by Trichoderma viride strain H06 using response surface methodology LIANG ChangCong, LIU Lei, ZHANG JianHua, WANG WeiWei, HUANG JunSheng﹡ (Environment and Plant Protection Institute，CATAS/ Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture / Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests，Haikou 571101) Abstract: The fermentation medium components for sporulation quantity by Trichoderma viride strain H06 was optimized by response surface methodology (RSM). Firstly, three of the most significant influence factors were screened by the method of Plackett－Burman design as corn meal, soybean meal and KH2PO4. The path of steepest ascent was applied to approach the optimal region of the three significant factors. Lastly, the optimal concentration and correlations among three factors were identified by RSM. The optimal fermentation conditions were determined as followed: sucrose10g/L, corn meal 12.75 g/L, NH4NO3 2 g/L, soybean meal 4.65 g/L, MgSO4·7H2O 1 g/L, KH2PO43.32 g/L. Under these conditions，the maximum theoretic spore number was 3.29×109 /mL. After three parallel verifications，the maximum theoretic value was consistent with mean value of verification test and the spore number was increased by 197% compare to that before optimization. Key words: RSM; Trichoderma viride; sporulation quantity; optimization 香蕉枯萎病是由尖孢镰刀菌引起的一种毁灭性的病害，已严重危害到香蕉生产。目前综合防控技术香蕉枯萎病防治的主要手段木霉（Trichoderma spp.）作为一种重要的植病生防菌一直受到普遍关注。能有效地竞争营养和生存空间,还可以产