摘要
生防细菌NCD-2是一株枯草芽孢杆菌菌株,该菌株通过分泌抑菌肽而对棉花黄萎病病原菌和棉花立枯病病原菌起到抑制作用。本研究着重通过原生质体法与诱导转座方法,建立了携带转座子Tn917质粒pTV1对枯草芽孢杆菌NCD-2野生菌株的转化体系与转座子突变技术,获得1500多个转座子插入突变子。通过测定这些突变子对大丽轮枝菌的抑制作用,筛选到2个抗生作用丧失的抑菌功能缺失的突变子。室内盆栽试验结果表明这2个抑菌功能缺失突变子对棉花立枯病的防效显著低于野生菌株,说明NCD-2野生菌株产生的抑菌肽在该菌株防治棉花立枯病中起到主要作用,进而说明编码该抑菌肽的基因在该菌株防治棉花立枯病中具有重要作用。
The bacterial isolate NCD-2,isolated from the rhizosphere of cotton, was an effective Bacillus subtilis strain to control cotton Verticilliurn wilt in field. Further it had been proved that this bacterium secreted antipeptides to inhibit the plant pathogens, Verticillium dahliae of cotton Verticillium wilt and Rhizoctonia solani of cotton sore shin. This paper determined the role of the antipeptide in controlling cotton disease. A mutagenesis technique system of B. subtilis NCD-2 was generated by transposon Tn 917 mutageneisis, and the functional gene encoding the antipeptide was knocked off by the transponson. In this study, B. subtilis NCD-2 was transformed with a plasmid pTV1 carrying transponson Tn917 by protoplast methods. Twenty tranformed NCD-2 strains were screened resistant against chloromycetin, erythromycin and lincomycin. By the transposon Tn917 mediated insertional mutagenesis technique, the transponson Tn 917 successfully inserted the genome of B.subtilis NCD-2 and 1500 mutants were sreened that were resistant to erythromycin and lincomycin but susceptible to chloromycetiru Among them, 2 antibiosis-free mutants were screened against V. dahliae. The results from the experiement on control of cotton sore shin with wild NCD-2 and antibios-free mutants in greenhouse showed that antibiosis of NCD-2 played an important role in controlling this disease with B. subtilis NCD-2. This suggested that the functional gene of B. subtilis NCD-2 encoding the antipeptide against V. dahliae had a significant function incontrol of cotton sore shin disease.
出处
《棉花学报》
CSCD
北大核心
2006年第3期131-134,共4页
Cotton Science
基金
河北省自然科学基金资助项目(C2004000735)
河北省农科院重点项目(A03-1-03-09)
