基于16SrDNA序列分析研究根腐病三七根内可培养细菌的多.PDFVIP

基于 16S rDNA 序列分析研究根腐病三七根内可培养细菌的多样性 张智慧 1，张倩茹 2，付晓萍 2，李凌飞 2* 1. 云南省农业科学院 药用植物研究所，云南 昆明 650223 2. 云南农业大学 食品科学技术学院，云南 昆明 650201 摘 要：目的 分析根腐病三七根内细菌的多样性。方法 用牛肉膏蛋白胨培养基分离根腐病三七根内的细菌，经细菌通用 引物 27F/1492R 扩增 16S rDNA 后，分别用 Rsa I 和 Hin6 I 限制性内切酶对扩增产物进行酶切，结合限制性片段长度多态性 （RFLP）分析方法和 DNA 测序技术，对分离自根腐病三七根内的细菌进行初步鉴定。结果 根腐病三七根内的细菌分属于 8 个类群，依占总菌数的比例分别是芽孢杆菌属 Bacillus 22.47%、无色杆菌属 Achromobacter 5.62%、寡养单胞菌属 Stenotrophomonas 5.62%、类芽孢杆菌属 Paenibacillus 4.49%、鞘氨醇杆菌属 Sphingobacterium 1.12%、苍白杆菌属 Ochrobactrum 1.12%、不动杆菌属 Acinetobacter 1.12%及肠杆菌科的一些属 58.43%。结论 泛菌属和芽孢杆菌属是根腐病三七根中的两大 优势细菌类群。 关键词：三七；根腐病；细菌；RFLP 分析；DNA 测序；16S rDNA 中图分类号：R282.12 文献标志码：A 文章编号：0253 - 2670(2014)03 - 0 - 05 DOI: 10.7501/j.issn.0253-2670.2014.03. Cultivable bacterial diversity in rotting roots of Panax notoginseng with 16S rDNA sequence analysis ZHANG Zhi-hui1, ZHANG Qian-ru2, FU Xiao-ping2, LI Ling-fei2 1. Institute of Medicinal Plant, Yunnan Academy of Agricultural Sciences, Kunming 650223, China 2. College of Food Science and Technology, Yunnan Agriculture University, Kunming 650201, China Abstract: Objective To analyse the bacterial diversity in rotting roots of Panax notoginseng. Methods Bacterial strains were isolated from the diseased roots of P. notoginseng using beef extract-peptone medium. The 16S rDNA was amplificated by primer 27F/1492R, the product was digested by restriction endonuclease Rsa I and Hin6 I, and PCR-RFLP analysis and DNA sequencing technology were used to identify the bacterial strains in rotting roots of P. notoginseng. Results The bacteria could be divided into eight groups including Bacillus (22.47%), Paenibacillus (4.49%), Sphingobacterium (1.12%), Ochrobactrum (1.12