一株番茄灰霉病菌拮抗菌的系统鉴定

Y-21是从山东省淄博市蔬菜温室中分离到的一株细菌，通过室内平板对峙试验，对番茄灰霉病菌具有较强的拮抗性。本研究通过形态观察、生理生化反应和分子生物学方法对这株细菌进行系统鉴定。显微镜观察菌株形态为杆状；有芽孢；革兰氏染色呈阳性。采用经典的基因组提取方法提取基因组DNA，以其为模板，利用引物63F、1494R扩增16SrRNA基因，回收16SrRNA基因的PCR产物并与T载体连接，通过热激法将重组质粒转人大肠杆菌感受态细胞，利用蓝白斑筛选，挑出白斑，提取质粒，通过蓝白斑质粒的电泳图比对和质粒的PCR检测确定阳性克隆，将阳性克隆菌株送样测序。将测序结果输入BLAST进行比对，发现Y-21属于假单胞菌属，然后对其构建系统发育树。

System identification of a Botrytis cinerea antagonistic bacteria

Y-21 has a stronger inhibition effect on Botrytis einerea. It was identified as Bacillus subtilis based on the results of morphologic characteristics, physiological and biochemical properties and phylogenetie analysis of 16S rDNA sequence. Y-21 is the rod-shaped bacteria, it is Bacillus, gram stain Y-21 is negative. To identify the bacteria by these genes. In this paper, a classic Genome extracting methods is used to extract the genome of the bacteria. Primers 63f, 1494r are used to amplify the gene of 16SrRNA the PCR products of 16SrDNA to connect with T-vector, and then transformating the recombinant plasmid into E. eoli competent cells, through blue-white selection, Singled out leukoplakia, then extract plasmid By the blue-white plasmid eleetrophoresis map＇ than the right and the plasmid was determined by PCR-positive clones. The positive clones are sequenced, The result of the sequences are imported by BLAST to compare their identification , find that Y-21 is Pseudomonas strain. And then construct phylogenetie trees to confirm the honmdgy of Y-21.