快速粗提取致病疫霉菌菌丝DNA的方法  

Rapid Coarse DNA Extraction Method for Phytophthora infestans Mycelia

作　　者：李海娟[1] 孙银银[1] 权军利[1] 单卫星[1] 

机构地区：[1]西北农林科技大学植物保护学院,陕西杨凌712100

出　　处：《北方园艺》2010年第17期168-170,共3页Northern Horticulture

基　　金：国家公益性行业科研专项资助项目(3-20);现代农业产业技术体系资助项目(nycytx-15)

摘　　要：为满足规模分析病菌群体、认识病菌的群体遗传变异特征,以致病疫霉菌(Phytoph-thorainfestans)为材料,建立快速、简便的用于PCR扩增的病菌基因组DNA。分别大量和微量提取基因组DNA,比较2种方法所提取的DNA用于SSR标记的PCR扩增效果。结果表明:以微量粗提法所得的病菌基因组DNA质量足以用于PCR扩增,与大量提取所得的DNA的PCR扩增非常一致,并且粗提的DNA与大提的DNA一样都可以在-20℃条件下保存。基于PCR扩增的遗传标记广泛用于病菌群体的分析,该研究建立的基于玻璃珠振荡法提取疫霉菌菌丝基因组DNA的微量提取方法需材少、简便且快速有效,为规模分析疫霉菌群体奠定了基础。To facilitate large scale analysis and understanding of genetic variation in pathogen populations using PCR-based molecular markers,aquick,simple procedure was developed for extracting genomic DNA of Phytophthorainfestans from small amount of mycelial tissues.Genomic DNA was extracted using urea-based large-scale procedure and the developed mini-scale protocol,and the quality was compared by PCR amplification of selected SSR markers.The results showed that there was no significant difference for PCR amplification of SSR markers between the genomic DNA isolated using two different methods,and the coarse DNA can also remain stable under-20℃.PCR-based molecular markers are widely used in population studies of plant pathogens,the developed glass beads-based method is simple,fast,and uses tiny amount pathogen mycelial tissues,making it possible for large-scale analysis of pathogen population and a better understanding of pathogen population biology.

关 键 词：疫霉菌 基因组DNA PCR扩增 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]