摘要
利用细菌16S rDNA基因的通用引物对16个供试菌株进行PCR扩增,把扩增产物进行核苷酸序列测定。将获得的序列与GenBank中相关菌株的16S rDNA序列进行同源性分析。以此设计出检测A.a.c的特异性引物,并利用最大简约法构建了16S rDNA系统演化树。系统演化关系分析表明,6～9号供试菌株的16S rDNA序列与A.a.c标准菌株仅有3个位点的差异,其同源性均在99.8%以上,在构建的系统演化树上,它们聚为同一个族群。利用设计的一对特异性引物(BFB64/65),对各供试菌株进行PCR检测,结果只有A.a.c相关菌株产生扩增条带,产物大小与预期一致。
Sixteen testing strains were amplified by polymerase chain reaction using the universal primers of the bacteria 16S rDNA gene, and the nucleotide sequences of their amplified fragment were tested. The homology between these sequences and the 16S rDNA sequence of related bacteria strains in GenBank was analyzed. A phylogenetic tree based on 16S rDNA sequences was constructed and showed that there were three nucleotides difference between the 16S rDNA sequences of No. 6 - 9 strains and the standard strain of A. a. c, the homology was more than 99.8%. They were clustered into the same phylogenetic branch in the tree. The result of PCR showed that only A. a. c strains came into being electrophoresis band. A. a. c was detected from different pathogenic germs, and a rapid and accurate method for detecting was established from molecular biology.
出处
《植物病理学报》
CAS
CSCD
北大核心
2007年第3期225-231,共7页
Acta Phytopathologica Sinica
基金
国家质量监督检验检疫总局科研项目(2004IK-029)
