乌江网箱养殖丁(魚歲)细菌的分离与分子鉴定

利用微生物学常用的细菌分离、纯化方法从丁(魚歲)中分离出9株菌株,编号分别为Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ、Ⅵ、Ⅶ、Ⅷ、Ⅸ.分别扩增出9株菌株的16S rRNA基因约600 bp的片段,并对其测序,将测序结果输入到NCBI(http://www.ncbi.nlm.nih.gov/)中,用BLAST工具对序列进行同源性比对分析,并利用MEGA4绘制系统进化树,结果表明:9株菌株的16S rRNA基因测序结果分别与不动杆菌(Acinetobacter)、嗜水气单胞菌(Aeromonas hydrophi-la)、希瓦氏菌(Shewanella baltica)、不动杆菌(Acinetobacter)、希瓦氏菌(Shewanella baltica)、霍乱弧菌(Vibriocholerae)、枯草芽孢杆菌(Bacillus subtilis)、杀鲑气单胞菌(Aeromonas salmonicida)、肺炎克雷伯氏菌(Klebsiellapneumoniae)的16S rRNA基因的核苷酸序列同源,同源性分别为96%,99%,99%,97%,98%,98%,99%,98%,98%.9株菌株的系统进化树明显分为3支,Ⅰ和Ⅳ聚为一支,Ⅶ独聚一支,Ⅱ、Ⅲ、Ⅴ、Ⅵ、Ⅷ、Ⅸ聚为一支,其中Ⅱ和Ⅷ相聚,Ⅲ和Ⅴ相聚后再与Ⅵ相聚,最后与Ⅸ相聚.

Isolation and Molecular Identification of Bacteria from Tinca tinca Linnaeus of Net Cage Culture in the Wujiang River

The present experiment was undertaken with the tranditional microbiology method to isolate the bacteria from Tinca tinca Linnaeus of net cage culture in Wujiang River and 9 strains were isolated. The numbers of 9 strains were Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅴ, Ⅵ, Ⅶ, Ⅷ, Ⅸ respectively and the 600bp fragments of 16S rRNA gene of nine strains were amplificated by PCR methold. These 600bp sequences were successfully sequenced and were entered into the NCBI (http://www. ncbi. nlm. nih. gov/). Objective to analyse the homology, BLAST tool was used and the Phylogenetic tree of the nine strains was constructed by the soft-ware of MEGA4. The results indicated that: the sequencing results of 16S rRNA gene of 9 strains have homology with the 16S rRNA gene nucleotide sequence of Acinetobacter, Aeromonas hydrophila, Shewanella baltica, Acinetobacter, Shewanella bahica, Vibrio cholerae, Bacillus subtilis, Aeromonas salmonicida, Klebsiella pneumoniae respectively and homology are as follows: 96%, 99%, 99%, 97%, 98%, 98%, 99%, 98%, 98%. The Phylogenetic tree of 16S rRNA gene of nine strains are clearly divided into three branches, in which Ⅰ and Ⅳ were together for one branch, Ⅶ are clustered alone and Ⅱ, Ⅲ, Ⅴ, Ⅵ, Ⅷ, Ⅸ were clustered together as another branch, thereinto, Ⅱ and Ⅷ are together, Ⅲ and Ⅴ were combined with Ⅵ and then togethered with Ⅸ.