玉米丝黑穗病菌DNA提取及UP-PCR反应体系建立

Extraction of DNA and Construction of UP-PCR Fingerprinting of Sporisorium reiliana

采用CTAB法、SDS法和尿素法,从玉米丝黑穗病菌液体培养物中提取基因组DNA,比较其提取效果。结果表明:改进的CTAB法可有效去除多酚、多糖和色素等杂质对DNA的污染,可获得高质量模板DNA。以此为模板进行UP-PCR(Universally Primed PCR)反应,对UP-PCR扩增反应中的一些重要参数进行了优化,建立适合于玉米丝黑穗菌的UP-PCR反应体系。

Three methods （CTAB, SDS and Urea）were adopted to isolate the genomic DNA of Sporisorium reiliana. The results showed that the reformed CTAB was more effective to remove most of polysaccharides and pigments, to inhibit the formation of oxidized polyphenolic compounds. Based on the genomic DNA extracted from liquid cultures of Sporisorium reiliana, essential factors affecting the results of UP-PCR（Universally Primed PCR） analysis were tested and compared. An optimal reaction system suited for UP-PCR in genetic diversity of Sporisorium reiliana was established.