棉花黄萎病生防内生芽孢杆菌LH-L3的分离鉴定  

Isolation and Characterization of Endophytic Bacillus methylotrophicus LH-L3 Antagonist of Cotton Verticillium Wilt

作　　者：陈丽华[1] 袁德超 吴毅歆[2,3] 何鹏飞 何月秋[2,3] 

机构地区：[1]云南农业大学植物保护学院,昆明650201 [2]云南农业大学农学与生物技术学院,昆明650201 [3]微生物菌种筛选与应用国家地方联合工程研究中心,昆明650217

出　　处：《棉花学报》2017年第6期550-559,共10页Cotton Science

基　　金：云南科技支撑计划(2014BB018)

摘　　要：【目的】获得能够防治黄萎病菌的棉花内生芽孢杆菌。【方法】将棉株组织研磨液80℃加热后,采用涂板法分离内生菌,将对大丽轮枝菌强致病力菌株Vd084拮抗效果最好的菌株,依据菌体形态、生理生化特征、16S rRNA、gyrB和rpoB基因序列分析进行鉴定,并以盆栽试验确定其对棉花黄萎病的防效。【结果】分离到61株芽孢杆菌类似菌,其中17株能拮抗大丽轮枝菌。复筛得到1株拮抗活性较高、抑菌谱较广的菌株LH-L3。该菌株被鉴定为甲基营养型芽孢杆菌。与无菌水对照相比,用106 mL^(-1)的LH-L3菌液浇灌3次的棉花出苗率、株高、根长、地上以及地下部分鲜物质质量分别提高42.85%、10.24%、23.83%、10.05%、97.62%,对棉花黄萎病的防治效果达到85.24%。【结论】菌株LH-L3有较好的促生长和棉花黄萎病防治的效果,具有进一步开发应用潜力。[Objective] The aim of this study is to obtain endophytic Bacillus with control effect on cotton Verticillium wilt.[Method] The cotton tissue grinding liquid was heated at 80 ℃, and spread onto the Luria-Bertani ager medium. The strains with better inhibitory effects on Verticillium dahliae Vd084, a strain with strong pathogenicity, was identified by the morphology,physiological and biochemical traits as well as 16 S rRNA, gyrB and rpoB sequences, and its control effect on cotton Verticillium wilt was determined by pot experiments. [Result] 61 similar Bacillus strains were isolated from cotton plants. Among them, 17 strains could inhibit Verticillium dahliae Vd084. A strain named LH-L3 with a relatively higher antagonistic activity and a broad inhibition spectrum was obtained in the second screening. LH-L3 was classified into Bacillus methylotrophicus. Compared with the control, the emergence rate, plant height, root length, fresh weight of aerial part and underground part of cotton with 106 mL^（-1） LH-L3 suspension drenching for 3 times increased by 42.85%, 10.24%, 23.83%, 10.05% and 97.62%, respectively. And control effect on Verticillium wilt was 85.24%. [Conclusion] LH-L3 had good growth promotion and Verticillium wilt control effects and showed good potential for biological control.

关 键 词：棉花 大丽轮枝菌 甲基营养型芽孢杆菌 防治效果 

分 类 号：S435.62[农业科学—农业昆虫与害虫防治]