毛竹枯梢病拮抗细菌分离鉴定及其拮抗物质  

Screening,Identification and Fermentation Conditions Optimization of Antagonistic Strain 6-59 of Bacillus megaterium against Ceratosphaeria phyllostachydis

作　　者：李潞滨[1] 李术娜[2] 李佳[2] 曾来涛[2] 胡陶[1] 朱宝成[2] 

机构地区：[1]中国林业科学研究院林业研究所国家林业局林木培育重点实验室,北京100091 [2]河北农业大学生命科学学院,保定071001

出　　处：《林业科学》2009年第7期63-69,共7页Scientia Silvae Sinicae

基　　金：“948”项目(2006-4-C08);国家“十一五”科技支撑计划课题(2006BAD01A18);北京市园林绿化局科技计划重点项目(2008-1)

摘　　要：从采集的土样中筛选到对毛竹枯梢病致病菌竹喙球菌具有拮抗作用的菌株78株,通过复筛选出1株拮抗活性较高的拮抗菌株6-59。通过形态观察、生理生化试验及16SrDNA的序列分析鉴定该菌株为巨大芽孢杆菌。经有机溶剂萃取和盐析法初步推断该菌株的拮抗物质为蛋白类物质。通过单因素试验和正交试验对菌株6-59摇瓶发酵条件进行优化,最终确定其最佳发酵条件为:蔗糖2%、蛋白胨3%、MgSO4.7H2O0.03%、KCl0.02%、种龄18h、培养基初始pH7.0,250mL三角瓶装液量50mL、接种量4%、发酵时间72h。在此条件下培养得到的菌株6-59的发酵液,其抑菌圈直径可达25.8mm。The 78 strains were collected from the soil and were prescreened to have antagonistic activity against Ceratosphaeria phyllostachydis. A strain named 6-59 with a relatively higher antagonistic activity was obtained via re-screening. Then the strain 6-59 was characterized by morphological and culture features observation, physiological and biochemical measurements and 16SrDNA sequence analysis. Based on the data, the strain was initially identified as Bacillus megaterium. Through organic solvent extracting and （NH4 ）2SO4 salting out to the fermentation liquid, the antagonistic substance produced by the stain 6-59 was inferred to a protein. Then the fermentation conditions of the strain 6-59 was studied via single factor experiments and orthogonal experiments, and the optimal shaking flask fermentation condition was determined as follows： a medium composed of 2% sucrose, 3% peptone, 0.03% MgSO4 ＂71-I20, 0.02% KCl, seed age 18 h, initial pH of the medium 7.0, medium 50 mL in a 250 mL flask, inoeulum size 4 % and fermentation time 72 h. In this optimized conditions, the inhibition zone diameter of the fermentation liquid could reach to 25.8 mm.

关 键 词：毛竹枯梢病 竹喙球菌 拮抗细菌 筛选 鉴定 拮抗物质 

分 类 号：S763.1[农业科学—森林保护学]