抗纹枯病、根腐病的转SN1基因小麦的获得与鉴定  

Development and Characterization of SN1 Transgenic Wheat Plants with Enhanced Resistance to Rhizoctonia cerealis and Bipolaris sorokiniana

作　　者：王金凤[1,2] 杜丽璞[1] 李钊[1] 黄素萍[1,2] 叶兴国[1] 冯斗[2] 张增艳[1] 

机构地区：[1]中国农业科学院作物科学研究所,农作物基因资源与基因改良国家重大科学工程/农业部作物遗传育种重点实验室,北京100081 [2]广西大学农学院,广西南宁530004

出　　处：《作物学报》2012年第5期773-779,共7页Acta Agronomica Sinica

基　　金：国家转基因生物新品种培育科技重大专项(2008ZX08002-001,2009ZX08002-006B)资助

摘　　要：SN1是源于马铃薯的一种抗菌肽,可以抑制多种植物病原菌的生长。小麦纹枯病(主要病原菌为禾谷丝核菌Rhizoctonia cerealis)和根腐病(主要病原菌为平脐蠕孢菌Bipolaris sorokiniana)是小麦的主要土传真菌病害。本研究利用基因工程技术构建了SN1基因的单子叶植物表达载体pA25-SN1,它受玉米泛素(ubiquitin)启动子的控制;采用基因枪法将pA25-SN1转化小麦推广品种扬麦18幼胚愈伤组织4000块,获得203株再生植株,通过PCR检测出阳性植株55株,转化率为1.38%。对外源SN1转基因小麦T0～T2代植株,进行目标基因的PCR、Southern blot、RT-PCR、荧光定量RT-PCR(Q-RT-PCR)分析和小麦纹枯病菌与根腐病菌接种及其抗病性鉴定。结果表明,转入的SN1基因已经整合到转基因小麦的基因组中,能够在转基因小麦中遗传、转录与表达。SN1基因的表达提高了转基因植株对小麦纹枯病和根腐病的抗性,其抗病性可以遗传。SN1 is an anti-microbial peptide in potato,and can inhibit the growth of important phytopathogens.In this study,the transformation vector of SN1 gene expressing highly in monocot plants,pA25-SN1,was correctly constructed,in which SN1 gene was driven by maize ubiquitin promoter.Four thousand embryo calli of Yangmai 18 were bombarded by the particle containing pA25-SN1.Among the regenerated 203 plants,55 positive transgenic individuals were identified by PCR assay in the T0 generation with a transformation frequency of 1.38%.The transgenic wheat plants from T0 to T2 generations were subjected to PCR,Southern blot,RT-PCR,and Q-RT-PCR assays.The disease responses of these positive transgenic plants were also evaluated by inoculating with Rhizoctonia cerealis and Bipolaris sorokiniana.The results showed that SN1 gene was integrated into these transgenic lines,inherited from T0 to T2,and expressed in the wheat background.The transgenic wheat plants expressing SN1 displayed an enhanced resistance to R.cerealis and B.sorokiniana compared with the untransformed Yangmai 18,and the enhanced resistance was inheritable.

关 键 词：SN1基因 转基因小麦 小麦纹枯病菌 小麦根腐病菌 抗性 

分 类 号：S512.1[农业科学—作物学]