ywbB基因对枯草芽胞杆菌NCD-2菌株生物膜形成和根际定殖能力的影响  

Functional analysis of ywbB gene to the biofilm formation and root colonization in Bacillus subtilis strain NCD-2

作　　者：郭庆港[1] 吴园园[1,2] 李社增[1] 鹿秀云[1] 王洪港 马平[1] 

机构地区：[1]河北省农林科学院植物保护研究所河北省农业有害生物综合防治工程技术研究中心农业部华北北部作物有害生物综合治理重点实验室,保定071000 [2]河北大学生命科学学院,保定071002 [3]河北省黄骅市城市管理局,黄骅061104

出　　处：《植物保护学报》2013年第1期45-50,共6页Journal of Plant Protection

基　　金：国家现代农业产业技术体系(CARS-18-15);国家自然科学基金(30900962;31272085);国家"863"计划(2011AA10A205)

摘　　要：为明确枯草芽胞杆菌Bacillus subtilis NCD-2菌株生物膜形成相关基因及其在生物膜形成、根际定殖中的作用,采用转座子插入技术,从4000余株NCD-2突变子中获得6株生物膜形成能力降低的突变子。在生物膜形成能力丧失的突变子M3中,转座子以单拷贝的形式插入到ywbB基因内部。通过基因同源重组技术对ywbB基因进行缺失突变,获得ywbB基因缺失突变菌株NCD-2(ΔywbB),与野生菌株相比,突变子丧失了生物膜形成能力。进一步比较发现,突变菌株NCD-2(ΔywbB)显著降低了在棉花根际的群体数量,同时也降低了对棉花立枯病的防治效果。研究证明,ywbB是NCD-2菌株生物膜形成途径中的重要基因,且生物膜形成与NCD-2菌株根际定殖和生防效果呈正相关。Bacillus subtilis NCD-2 is a plant beneficial bacterium which forms biofilm in vitro and in plan- ta. In order to elucidate the role played by biofilm formation and its associated gene （s） in the rhizo- sphere colonization and bio-control capability of Bacillus subtilis NCD-2, a Tn-lO transposon mutant li- brary of strain NCD-2 was constructed in this study. Six out of 4 000 mutants showed deficiency in biofilm formation. The mutant strain M3, which was dramatically lost the capability of biofilm formation, was fur- ther studied. Gene ywbB was identified which was inserted by the transposon Tn-lO with a single copy in mutant strain M3. A deletion mutant of ywbB, mutant strain NCD-2 （AywbB）, was generated to charac- terize the role of ywbB in strain NCD-2. Compared with the wild strain NCD-2, mutant strain NCD-2 （AywbB） also showed deficiency in biofilm formation. Meanwhile, mutant strain NCD-2 （AywbB） de- creased the ability of colonization in cotton rhizosphere and control against cotton seedling rhizoctoniadamping-off. The results confirmed that ywbB was an strain NCD-2. It was suggested that biofilm formation tion and disease control of strain NCD-2. important gene among biofilm was positively correlated with formation pathway in rhizosphere coloniza-

关 键 词：枯草芽胞杆菌 生物膜 定殖 转座子 

分 类 号：S476[农业科学—农业昆虫与害虫防治]