拮抗南方根结线虫PKS基因的筛选及活性评价  

Screening for PKS gene from soil metagenomics library and identification of the actives against root-knot nematodes

作　　者：罗坤[1,2] 赵志祥[1,2] 芦晓飞[2] 黎定军[1,3] 谢丙炎[2] 

机构地区：[1]湖南农业大学生物安全科技学院,长沙410128 [2]中国农业科学院蔬菜花卉研究所,北京100081 [3]湖南广播电视大学,长沙410004

出　　处：《植物保护》2010年第5期38-42,共5页Plant Protection

基　　金：国家重点基础研究发展计划(2009CB119000);公益性行业(农业)科研专项经费(3-25,nyhyzx07-050);“十一五”国家科技支撑计划(2006BAD07B03,2006BAD08A08,2006BAD17B08)

摘　　要：土壤环境中蕴藏着巨大的微生物资源,而99%的土壤微生物不能用传统的纯培养。本研究基于西藏米拉山高寒草甸土壤微生物宏基因组Fosmid文库,采用PCR方法从文库中筛选PKS基因,以南方根结线虫(Meloido-gyne incognita)为靶标测定其杀线活性,并验证温室盆栽防效。从Fosmid文库中筛选得到2个含PKS基因的克隆K99和K82。K99属于Ⅰ型PKS,K82属于杂合PKS/NRPS;K99发酵液、热处理发酵液浸泡南方根结线虫J2 12 h后,杀线活性为100%。温室盆栽防效结果表明,K99的发酵菌液对南方根结线虫的防效为89%。K99能产生一种外分泌有杀线活性的物质,其热稳定性好,具有重要的开发利用价值。Microorganisms dominate the soil ecosystem, but their roles are unclear due to the limitations associated with the uncultivability of at least 99% of the microorganisms. Clones containing PKS KS domain were screened by PCR method from a soil Fosmid metagenomic library from Tibet Mila Mountain, and the actives of PKS clones against root-knot nematodes were also identified according to control efficiency in plots in greenhouses. The re- sults showed that two clones （K99 and K82） containing PKS were obtained, of which K99 was type I PKS and K82 the hybrid PKS/NRPS. The control effects of crude zymotic fluid and heat treatment fermentation liquid of K99 clone on J2 of root-knot nematode in 12 hours was 100%, and the control efficiency in plots in greenhouses to nematode was 89%. Clone K99 produced property of thermal stability. Therefore it some active substances that could kill root-knot nematode and had a was well worth exploiting and utilizing

关 键 词：PKS 南方根结线虫 杀线活性 土壤宏基因组文库 

分 类 号：S476[农业科学—农业昆虫与害虫防治]