食用菌木霉的生防细菌鉴定及相关基因功能预测  

Identification of an antifungal bacterium against mushroom pathogen Trichoderma viride and characterization of genes associated with antifungal activity

作　　者：马林[1,2] 曲绍轩[1] 王晓强[3] 邓鹏[4] 林金盛[1] 李辉平[1] 侯立娟[1] 蒋宁[1] 宋金俤[1] LU Shi-en 

机构地区：[1]江苏省农业科学院蔬菜研究所,江苏南京210014 [2]江苏省高效园艺作物遗传改良重点实验室,江苏南京210014 [3]山东农业大学植物保护学院,山东泰安271018 [4]美国密西西比州立大学生物化学分子生物学昆虫学与植物病理学学院,美国密西西比州39762

出　　处：《江苏农业学报》2016年第3期528-533,共6页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省青年基金项目[BK20150547];现代农业产业技术体系建设专项资金(CARS24)项目

摘　　要：采用抑菌圈法筛选出12株对绿色木霉有明显抑制作用的生防细菌,其中抑菌效果较好的为12C2-4和MS82菌株。通过对峙培养法发现MS82菌株对双孢蘑菇菌丝生长几乎没有影响。通过API20 NE、API50 CH生理生化指标测定及基于16S r DNA基因片段的系统进化分析对MS82菌株进行分类鉴定,结果显示MS82属于假单胞菌(Pseudomonas sp.)。为了明确MS82中参与抗真菌活性相关的基因,利用Tn5转座子随机插入的方式构建MS82菌株的突变体库,获得完全失去绿色木霉抗性的突变体MS82MT31。通过对抗菌失活突变体中突变基因定位,发现突变基因为rpo B基因,与已知菌株Pf0-1的rpo B基因相似性为96%,说明rpo B基因与MS82抑菌活性物质合成密切相关。To find soil antifungal strains against Trichoderma,the first competitive contaminant in mushroom cultivation,12 isolates of biocontrol bacteria inhibiting the growth of Trichoderma viride significantly were selected using inhibition zone method. Strains 12C2-4 and MS82 presented distinct antifungal activities against T. viride,among which,strain MS82 showed no inhibition against the mycelial growth of Agaricus bisporus by confronting incubation. Biological and biochemical characteristics of strain MS82 were identified by API20 NE and API50 CH systems. Together with the phylogenetic analysis of 16 S r DNA gene of MS82,it was showed that strain MS82 is a member of Pseudomonas sp..To characterize the genes of MS82 dedicated to antifungal activity,a mini MS82 mutant library was constructed using Tn5 transposome kit. Mutant MS82MT31 was defective in antifungal activity against Trichoderma sp. completely. BLAST analyses revealed that Tn5 transposon was inserted into the rpo B gene which shared a high similarity（ 96%） to the rpo B gene of P. fluorescens Pf0-1（ Gen Bank accession no. CP000094. 2）. These results suggested that rpo B gene is closely associated with the synthesis of antifungal compounds of MS82 strain.

关 键 词：食用菌 绿色木霉 拮抗细菌 抑菌活性 假单孢菌 

分 类 号：S482.292[农业科学—农药学]