番茄煤霉病生防融合子基因组RAPD分析鉴定

RAPD Analysis and Study of Biocontrol efficacy on Fusants against Tomato Cercospora leaf mold

RAPD分子标记以其快速、简便等优点,克服了传统的标记手段和形态分类学的缺点,在亲本和杂交种的鉴别以及在基因克隆与分离和遗传图谱的建立等研究中都起着重要的作用。以番茄煤霉病生防菌株木霉菌T-23和链霉菌A的融合子为实验材料,比较和研究了真菌融合子基因组DNA的提取。并用20个随机引物对亲本及其融合子进行RAPD分析。结果表明,SDS-CTAB法提取的基因组DNAOD260/OD280为1.909,DNA浓度约为42.0 ng/μL,可以满足分子生物学实验的需要;融合子是双亲融合的产物,但从双亲获得的遗传信息不等,融合子在DNA水平上更接近链霉菌。

RAPD molecular marker was widely applied to the studies of species and parental strain identification,genetic diversity,gene clone,gene isolation and the construction of gene linkage map due to its rapidity,simplicity and economy.Protoplast fusants were conducted from the strain of Streptonyce A and the strain of Trichoderma harzianum T-23.In the study,three methods were used to extract gDNA of fusants in order to find the best method of DNA extraction.The results showed that SDS-CTAB method was fit for extracting gDNA of fusants and the extraction could be directly used in RAPD-PCR amplification without any further purification.The OD260/OD280 value of the DNA is 1.909,concentration of the DNA is 42.0 ng/μL.The result illustrated that molecular biological evidence of protoplast fusion was obtained in biocontrol fungi by RAPD method in this experiment for the first time,we suggested that RAPD molecular marker is a valuable genetic marker for fungi study.