1种小麦白粉病菌DNA基因组的微量简捷提取方法

Simple and Rapid Method for DNA Genome Micro-Extraction fromWheat powdery mildew(Blumeria graminis f. sp. tritici)

为了寻找适合小麦白粉菌基因组DNA微量提取的方法,分别采用改进破壁法,液氮研磨法和溶菌酶消化法进行破壁,提取专性寄生菌小麦白粉菌DNA。结果表明,用改进的破壁方法,仅用3~10 mg的分生孢子粉所获得DNA的收率为(12.23±3.46)^(40.32±5.67)ng/mg,且OD260/OD280比值为1.71~1.92之间,说明该破壁方法获得的DNA收率大且纯度高。通过PCR反应获得了良好的效果。同时该方法也适用于小麦条锈菌和大麦白粉菌专性寄生菌DNA的提取。

Three different DNA extraction methods for wheat powdery mildew（Blumeria graminis f.sp.tritici） were adopted.They were modified cell-wall breaking method,liquid nitrogen grinding method,and lysozyme digestion method.The results showed that the recovery rate of DNA extracted with modified cell-wall breaking method was（12.23±3.46）^（40.32±5.67） ng/mg using only 3~10 mg of condia,and OD260/OD280 purity ratios were 1.71~1.92 indicated that cell-wall breaking method gained greater DNA yield and purity.PCR reactions obtained also good effects.At the same time,the method was also applicable to extract DNA from wheat stripe rust and barley powdery mildew and other DNAs from autoecious parasitic fungi.