盐芥T-DNA插入突变体库的建立及初步分析  

Construction of T-DNA Insertion Mutants Library of Thellungiella halophila and Its Preliminary Analysis

作　　者：王莹[1,2] 任大明[1] 

机构地区：[1]沈阳农业大学,沈阳110161 [2]中国农业科学院,北京100081

出　　处：《湖北农业科学》2011年第4期845-848,共4页Hubei Agricultural Sciences

基　　金：国家973计划项目(2006CB101706)

摘　　要：以盐芥[Thellungiella halophila(C.A.Mey.)O.E.Schulz]为材料,利用根癌农杆菌(Agrobacteriumtumefaciens)介导的转化方法,获得T-DNA插入突变群体,构建了一个具有5 000株抗Basta/Kan的盐芥T-DNA插入突变体库,其中有若干个株系有明显的表型改变,包括花期的改变、株型矮小、叶片形状改变等。并优化了建库的各因素,确定了最佳的盐芥栽培技术及转化体系,盐芥春化温度及时间为4℃、30 d,转化用菌液浓度OD600≈2.0,侵染时间2 min,暗培养1 d,Kan筛选浓度50 mg/L,除草剂(Basta)筛选浓度为稀释1 500倍;这些因素可为根癌农杆菌介导的盐芥转基因、突变体构建等基因组学研究提供强有力的技术平台。An T-DNA insertion mutant library containing 5 000 independent Basta/Kan-resistant lines was constructed by Agrobacterinm tumefaciens-mediated transformation using Thellungiella halophila（C.A.Mey.） O.E.Schulz as material.Among these transformations,several lines showed obvious phenotypes different from wild-type,including late flowering time,dwarf,changed leaf shape and so no.And the optimization of the conditions for Agrobacterium-mediated gene transformation of T.halophila was discussed.The optimal conditions were as follows： bacterial concentration OD600≈2.0,infection time 2 min,dark culture 1 d,kana mycin concentration 50 mg/L,basta were diluted to 1 500 fold.In addition,the results of the study on T.halophila vernalization showed that the bolting time was 30 days,vernalization temperature range was 4～5℃.This was an efficient and powerful technique for genomic studies on its transformation,mutant construction and so on.

关 键 词：T-DNA 盐芥 突变体 

分 类 号：Q754[生物学—分子生物学] Q949.748.3