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作物学报 ›› 2012, Vol. 38 ›› Issue (05): 773-779.doi: 10.3724/SP.J.1006.2012.00773

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

抗纹枯病、根腐病的转SN1基因小麦的获得与鉴定

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,*   

  1. 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
  • 收稿日期:2011-11-08 修回日期:2012-01-19 出版日期:2012-05-12 网络出版日期:2012-03-05
  • 通讯作者: 张增艳, E-mail: zhangzy@mail.caas.net.cn
  • 基金资助:

    本研究由国家转基因生物新品种培育科技重大专项(2008ZX08002-001, 2009ZX08002-006B)资助。

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

WANG Jin-Feng1,2,DU Li-Pu1,LI Zhao1,HUANG Su-Ping1,2,YE Xing-Guo1,FENG Dou2,ZHANG Zeng-Yan1,*   

  1. 1 National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of Crop Genetic and Breeding, Ministry of Agriculture / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2 College of Agronomy, Guangxi University, Nanning 530004, China
  • Received:2011-11-08 Revised:2012-01-19 Published:2012-05-12 Published online:2012-03-05
  • Contact: 张增艳, E-mail: zhangzy@mail.caas.net.cn

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1302

被引次数

10

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4 000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%。对转SN1基因小麦T0~T2代植株, 进行外源基因的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。

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

Abstract: 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 ubiquitinpromoter. 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 cerealisand 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 anenhanced resistance to R. cerealis and B. sorokiniana compared with the untransformed Yangmai 18, and the enhanced resistance was inheritable.

Key words: SN1 gene, Transgenic wheat, Rhizoctonia cerealis, Bipolaris sorokiniana, Resistance

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