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1种小麦白粉病菌DNA基因组的微量简捷提取方法
引用本文:龚双军,杨立军,刘辉,向立波,喻大昭.1种小麦白粉病菌DNA基因组的微量简捷提取方法[J].微生物学杂志,2011,31(1):24-27.
作者姓名:龚双军  杨立军  刘辉  向立波  喻大昭
作者单位:1. 湖北省农科院植保土肥研究所,湖北省重大农作物病虫草害重点实验室,湖北,武汉,430064
2. 襄阳市植保站,湖北,襄阳,441000
基金项目:公益性行业(农业)科研专项,国家小麦产业体系项目,公益性行业(农业)科研专项
摘    要: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。

关 键 词:小麦白粉病菌  DNA微量提取

Simple and Rapid Method for DNA Genome Micro-Extraction from Wheat powdery mildew(Blumeria graminis f.sp.tritici)
GONG Shuang-jun,YANG Li-jun,LIU Hui,XIANG Li-bo,YU Da-zhao.Simple and Rapid Method for DNA Genome Micro-Extraction from Wheat powdery mildew(Blumeria graminis f.sp.tritici)[J].Journal of Microbiology,2011,31(1):24-27.
Authors:GONG Shuang-jun  YANG Li-jun  LIU Hui  XIANG Li-bo  YU Da-zhao
Institution:GONG Shuang-jun1,YANG Li-jun1,LIU Hui2,XIANG Li-bo1,YU Da-zhao1(1.Inst.for Plant Protect'n & Soil Sci.,Hubei Acad.of Agric.Sci.,Hubei Key Lab.of Crop Dis.,Insect Pests & Weeds Control,Wuhan 430064,2.Plant Protection Station,Xiangyang City,Hubei Xiangyang 441000)
Abstract: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 310 mg of condia,and OD260/OD280 purity ratios were 1.711.92 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.
Keywords:wheat powdery mildew  DNA microextraction  
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