豌豆终止子rbc-T在转基因小麦研究中的应用  

Application of Pisum sativum Terminator rbc-T in Wheat Transformation

作　　者：王小婷[1,2] 黄锁 徐兆师[2] 李连城[2] 马有志[2] 陈明[2] 闵东红[1] WANG Xiao-Ting;HUANG Suo;XU Zhao-Shi;LI Lian-Cheng;MA You-Zhi;CHEN-Ming;MIN Dong-Hong(College of Agronomy, Northwest A&F University / State Key Laboratory of Crop Stress Biology for Arid Areas, Yangling 712100, China;National Key Facility for Crop Gene Resource and Genetic Improvement / Key Laboratory of Biology and Genetic Improvement of Triticeae Crops, Ministry of Agriculture / Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China)

机构地区：[1]西北农林科技大学农学院/旱区作物逆境生物学国家重点实验室,陕西杨凌712100 [2]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/农业部麦类生物学与作物遗传育种重点实验室,北京100081

出　　处：《作物学报》2017年第8期1254-1258,共5页Acta Agronomica Sinica

基　　金：国家转基因生物新品种培育重大专项(2014ZX08002-003B;2016ZX08002002;2016ZX08002005)资助~~

摘　　要：目前常用的转基因载体元件大多为真菌或细菌来源,引发对转基因食品安全性的担忧。本研究克隆了豌豆终止子rbc-T,并用其替换农杆菌终止子nos-T构建转化载体,利用基因枪法将携带Ubi启动子、GUS基因和终止子rbc-T组成的最小表达框片段转化普通小麦,同时以带有nos-T终止子的载体为对照,经过PCR检测筛选获得T2代稳定遗传的转基因小麦株系。GUS组织化学染色及酶活定量分析结果表明,带有nos-T及rbc-T的转基因小麦中均能检测到GUS基因不同程度的表达。因此认为,豌豆终止子rbc-T可以代替农杆菌终止子nos-T用于小麦的转基因研究。The fungal-or bacterial-origin vector elements are commonly used in transgenic crops,which raise great concernsabout the safety of GM foods.In the study,we isolated the pea(Pisum sativum)terminator,rbc-T,and constructed a transformationvector by replacing the Agrobacterium terminator nos-T with rbc-T.The minimum transformation fragment consisting ofUbiquitin promoter,GUS reporter gene and terminator rbc-T was transformed into common wheat by particle bombardmentmethod.Meanwhile a minimum transformation fragment with the Agrobacterium terminator nos-T was used as the control.The T2transgenic lines were regenerated after PCR validation of the transformation.In the transgenic lines carrying both kinds of terminators,GUS gene expression was detected by GUS histochemical staining and enzyme activity analysis,indicating that the peaterminator rbc-T is applicable as an alternative of the Agrobacterium terminator nos-T in transgenic wheat research.

关 键 词：胭脂碱合酶终止子(nos-T) 豌豆终止子(rbc-T) 转基因小麦 GUS报告基因 

分 类 号：S512.1[农业科学—作物学]