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作物学报 ›› 2017, Vol. 43 ›› Issue (08): 1254-1258.doi: 10.3724/SP.J.1006.2017.01254

• 研究简报 • 上一篇    

豌豆终止子rbc-T在转基因小麦研究中的应用

王小婷1,2,黄锁2,徐兆师2,李连城2,马有志2,陈明2,*,闵东红1,*   

  1. 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
  • 收稿日期:2017-01-20 修回日期:2017-04-20 出版日期:2017-08-12 网络出版日期:2017-04-27
  • 通讯作者: 闵东红, E-mail: mdh2493@126.com, Tel: 13609123593; 陈明, E-mail: chenming02@caas.cn, Tel: 010-82108750
  • 基金资助:

    本研究由国家转基因生物新品种培育重大专项(2014ZX08002-003B, 2016ZX08002002, 2016ZX08002005)资助。

Application of Pisum sativum Terminator rbc-T in Wheat Transformation

WANG Xiao-Ting1,2,HUANG Suo2,XU Zhao-Shi2,LI Lian-Cheng2,MA You-Zhi2,CHEN-Ming2,*,MIN Dong-Hong1,*   

  1. 1College of Agronomy, Northwest A&F University / State Key Laboratory of Crop Stress Biology for Arid Areas, Yangling 712100, China; 2 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
  • Received:2017-01-20 Revised:2017-04-20 Published:2017-08-12 Published online:2017-04-27
  • Contact: Min Donghong, E-mail: mdh2493@126.com, Tel: 13609123593; Chen Ming, E-mail: chenming02@caas.cn, Tel: 010-82108750
  • Supported by:

    This study was supported by the National Major Project for Developing New GM Crops (2014ZX08002-003B, 2016ZX08002002, 2016ZX08002005).

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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。

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, 报告基因

Abstract:

The fungal- or bacterial-origin vector elements are commonly used in transgenic crops, which raise great concerns about the safety of GM foods. In the study, we isolated the pea (Pisum sativum) terminator, rbc-T, and constructed a transformation vector by replacing the Agrobacterium terminator nos-T with rbc-T. The minimum transformation fragment consisting of Ubiquitin promoter, GUS reporter gene and terminator rbc-T was transformed into common wheat by particle bombardment method. Meanwhile a minimum transformation fragment with ?the Agrobacterium terminator nos-T was used as the control. The T2 transgenic 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 pea terminator rbc-T is applicable as an alternative of the Agrobacterium terminator nos-T in transgenic wheat research.

Key words: Nopaline synthase terminator (nos-T), Pea terminator (rbc-T), Transgenic wheat, GUS reporter gene

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