基于免培养技术的食用菌病害微生物rDNA测序及初步分析

rDNA Sequencing and Analysis of Edible Fungus Disease Microorganisms Using Culture-Independent Method

采用CTAB结合DNA凝胶回收试剂盒法提取四种主要食用菌病害组织样品的宏基因组DNA,用16S rDNA和18S rDNA通用引物分别对样品的基因组DNA进行PCR扩增、连接、转化并进行单克隆测序。每对引物随机挑取20个阳性克隆的rDNA序列进行比对,并对病害微生物的亲缘关系进行了初步分析,结果表明四种病害样品中两种为细菌性病害,两种为真菌性病害。

The microorganism metagenome of four kinds of edible fungus diseases was extracted by CTAB method combined with DNA gel recovery kit. The genomic DNA was amplified respectively by polymerase chain reaction using the universal primers of the 16S rDNA and 18S rDNA gene, and then monoclonal se- quenced after ligated and transformed. At least 20 positive clones selected by random of each pair of primers were performed sequence alignment. The results showed that there were two bacterial diseases and two fungaldiseases in the samples.