番茄灰霉病生防融合子基因组DNA提取方法的研究

Study on DNA Extraction Methods in Fusants against Botrytis Cinerea Pers on Tomato

以番茄灰霉病生防菌株木霉T-23和链霉菌A的融合子为实验材料,在SDS-CrAB法、改进CTAB法和氯化苄法的基础上加以改进,比较和研究了真菌融合子基因组DNA的提取,找到了一种快速、高效的基因组DNA提取方法,为进一步对融合子进行生防机制和分子生物学水平的研究提供基础。结果表明SDS-CTAB法提取的基因组DNA OD_(260)/OD_(280)为1．909,DNA浓度约为42．0ng/μL,可以满足分子生物学实验的需要。并将提取的基因组DNA直接用于PCR扩增,得到了多态性的RAPD图谱。

Protoplast fusants were conducted from the strain of Streptonyce A and the strain of Harzianum Tridhoderma T- 23. In the study,SDS - CTAB, modified CTAB and PhCH2C1 methods were used to extract gDNA of fusants in order to find the best method of DNA extraction.The results of the experiments showed that SDS - CTAB method was fit for extracting gDNA of fusants and they could be directly used in RAPD - PCR amplification without any further purification. The ratio of their OD260/OD280 value of the DNA is 1.909,concentration of the DNA is 42.0 ng/μL.