稻曲病菌基因组DNA提取方法比较与小文库构建  

Comparing the Methods of Isolating Genomic DNA and Construction of Its Small Genomic Library of Ustilaginoidea virens

作　　者：伏荣桃[1] 王剑[1] 陈诚[1] 龚学书[1] 卢代华[1] 罗曦[1] 郑爱萍[2] FU Rong-tao;WANG Jian;CHEN Cheng;GONG Xue-shu;LU Dai-hua;LUO Xi;ZHENGAi-ping(Key Laboratory oflntegrated Pest Management on Crops in Southwest, Ministry of Agriculture Science, Institute of Plant Protection, Sichuan Academy of Agriculture Science, Chengdu 610066;Rice Research Institute, Sichuan Agricultural University, Chengdu 611130)

机构地区：[1]四川省农业科学院植物保护研究所农业部西南作物有害生物综合治理重点实验室,成都610066 [2]四川农业大学水稻研究所,成都611130

出　　处：《生物技术通报》2018年第4期102-106,共5页Biotechnology Bulletin

基　　金：四川省财政创新能力提升工程项目(2016GYSH-014);四川省农业科学院青年基金项目(2015JSCX-017)

摘　　要：水稻稻曲病是由稻曲病菌引起的真菌病害,现已成为水稻重要的病害之一。旨在寻找一种最佳的稻曲病菌基因组DNA提取方法,并构建基因组小文库。采用CTAB、SDS和真菌DNA试剂盒3种提取方法提取稻曲病菌基因组DNA,并用Illumina系统测序分析,构建基因组小文库。结果显示,CTAB提取法能得到高质量的基因组DNA,小文库测序组装共得29 350288碱基(bp)和17 908 Scaffold,总碱基的GC含量为49.79%。CTAB提取法是稻曲病菌基因组DNA最佳的提取方法,基因组小文库成功的构建为稻曲病菌的基因组gap的填补和致病相关基因的鉴定提供了数据资源。Rice false smut caused by Ustilaginoidea virens is one of important fungal diseases for rice. This experiment aimed to select anoptimal method of isolating genomic DNA,and to construct small genomic library of U. virens. The genomic DNA of U. virens was extracted by 3 methods of CTAB,SDS,and fungal DNA isolation kit. The DNA sample was sequenced using Illumina system,and small genomic library wasconstructed. The results showed that the high-quality genomic DNA was obtained by CTAB. Meanwhile,the small genomic library of U. virenswas contained 29 350 268 bases and 17 904 Scaffold,and the GC content was 49.79% of total bases. We found that CTAB was the best methodof isolating genomic DNA of U. virens. The successful construction of the small genomic library provided data resource for filling the genome gapand identifying pathogenetic gene.

关 键 词：稻曲病菌 CTAB SDS 基因组小文库 

分 类 号：S435.111.4[农业科学—农业昆虫与害虫防治]