烟草青枯病生防菌黑曲霉发酵条件优化研究  

Optimization of fermentation condition of an Aspergillus niger ctrain against tobacco Ralstonia solanacearum

作　　者：丛铭[1] 万科[1] 禹辉[1] 陈雪 郭博恺 费长飞 王嫱[1] 李祝[1] 

机构地区：[1]贵州大学生命科学学院,贵州贵阳550025 [2]贵州省烟草公司毕节市公司,贵州毕节551700

出　　处：《广东农业科学》2014年第15期70-75,88,共7页Guangdong Agricultural Sciences

基　　金：贵州省烟草公司毕节市公司科技项目(201106);贵州省农业攻关科技计划项目(黔科合NY字[2011]3053);贵州省科学技术基金(黔科合J字[2010]2065号);贵州大学SRT项目(贵大SRT字[2013]082号)

摘　　要：以烟草青枯菌Ralstonia solanacearum为指示菌,黑曲霉Aspergillus niger对其的抑菌圈直径为响应值,运用单因素试验、Plackett-Burman(PB)设计、Box-Behnken(BBD)设计及响应面Response Surface Methodology(RSM)分析将分离并鉴定的黑曲霉菌株的液体发酵条件进行了优化。通过单因素试验确定麦芽糖和麦麸分别为最适碳源和氮源,通过Plackett-Burman设计对培养基中相关影响因素的效应进行评价,并筛选出有显著正效应的MgSO4及有显著负效应的CaCl2和吐温80等3个因素,使用最陡爬坡路径逼近最大抑菌圈直径,最后用BBD设计及RSM分析确定了主要影响因素的最佳条件。结果显示,经过优化的培养基组分为麦芽糖25.00 g/L、MgSO40.38 g/L、K2HPO40.10 g/L、KCl 0.50 g/L、CaCl20.05g/L、FeSO40.20 g/L、麦麸8.50 g/L、酵母膏1.50 g/L、吐温80 0.46 mL/L。采用滤纸片扩散法检测优化后的发酵液抗菌活性,抑菌圈直径为37.49(±0.34)mm,比未经优化测得的抑菌圈直径24.79(±0.72)mm增大了51.23%。The condition for liquid fermentation of an Aspergillus niger strain was optimized by using Design-Expert software combined with the Box-Behnken Design（BBD） and Response Surface Methodology（RSM） analysis method,and the Ralstonia solanacearum was used as indicator and inhibition zone Aspergillus niger aganst was used as the response value.The optimum carbon and nitrogen for Aspergillus niger were obtained through single factor experiment,which were maltose and wheat bran,respectively.Through Plackett-Burrman design,some factors of fermentation medium were evaluated and three factors had prominent effect on inhibition zone,which were MgSO4,CaCl2 and Tween 80,respectively.Then steepest ascent was used to approach maximum inhibition zone diameter.The BBD and RSM were used to determine the optimal levels of the main influencing factors,as the composition of fermentation medium was optimized as follow：maltose 25.00 g/L,MgSO40.38 g/L,K2HPO40.10 g/L,KCl 0.50 g/L,CaCl20.05 g/L,FeSO4 0.20 g/L,wheat brain 8.50 g/L,yeast paste 1.50 g/L,Tween 80 0.46 mL/L.Under those optimal conditions,the diameter of inhibitory zone increased by51.23% from 24.79（±0.72） to 37.49（±0.34） mm.

关 键 词：青枯菌 黑曲霉 发酵 PLACKETT-BURMAN设计 响应面法 

分 类 号：Q939.9[生物学—微生物学]