【摘要】 比较转Bt基因抗虫棉与其母本茎部内生细菌多样性,为评估转基因棉花生物安全性提供基础数据。采用平板培养方法对转基因棉花(中棉30,TC)与其母本(中棉16,CC)茎部内生细菌进行分离与计数,用细菌的通用引物扩增细菌的16SrDNA片段,进行RFLP分析。采用培养方法分离与计数所得的棉花茎部内生菌数量为105 cfu/g;随机挑取100个菌落,分别用限制性内切酶Hha I和Ras I酶切,转基因棉花中棉30(TC)茎部内生细菌酶切后产生10种OTUs,常规棉中棉16(CC)茎部内生细菌经酶切后产生9种OTUs;测序结果表明,中棉30(TC)优势菌属于Microbacterium和Curtobacterium属,中棉16(CC)优势菌为Curtobacterium和Enterobacter属。转Bt基因抗虫棉中棉30(TC)对茎部内生细菌数量和种类没有造成有害影响。更多还原

【Abstract】 This paper tries to compare the diversities of endophytic bacteria from Bt transgenic cotton and its parent cotton,and provides basic data for the biosafety evaluation of Bt transgenic cotton.Endophytic bacteria were isolated separately from the stems of two cottons(Zhongmian 30,Bt transgenic cotton,TC;Zhongmian 16,contrast non-transgenic parent,CC) by culture-dependent method.16S rDNAs were amplified with universal primers of bacteria,and PCR products were digested with restriction endoenzyme Hha I and Rsa I respectively,than analyzed by RFLP analysis.The results showed that endophytic bacteria content was 105 cfu/g.There were 10 OTUs(operational taxonomic units) in the stems of Bt transgenic cotton and 9 OTUs in contrast parent cotton.The 16S rDNA sequences of 12 respective strains were identified and submitted to GenBank.16S rDNA sequencing indicated that the dominant genera of Bt transgenic cotton are Microbacterium and Curtobacterium.The predominance families of corresponding receptor cotton are Curtobacterium and Enterobacter.Bt transgenic cotton did not bring harmful effects upon the number and diversity of endophytic bacteria in stems.更多还原