生物技术通报 ›› 2016, Vol. 32 ›› Issue (1): 103-108.doi: 10.13560/j.cnki.biotech.bull.1985.2016.01.017

• 研究报告 • 上一篇    下一篇

甘蔗一次多基因遗传转化及多重PCR检测

王文治, 杨本鹏, 蔡文伟, 熊国如, 王俊刚, 冯翠莲, 张树珍   

  1. 中国热带农业科学院甘蔗研究中心 中国热带农业科学院热带生物技术研究所 农业部热带作物生物学与遗传资源利用重点实验室, 海口 571101
  • 收稿日期:2015-03-27 出版日期:2016-01-09 发布日期:2016-01-22
  • 作者简介:王文治, 男, 硕士研究生, 助理研究员, 研究方向:甘蔗基因工程;E-mail:wangwenzhi@itbb.org.cn
  • 基金资助:
    国家自然科学基金项目(31371687), 现代农业产业技术体系建设专项资金项目(CARS-20-2-5)

Multiple Gene Transformation in Sugarcane and Multiple PCR Detection

WANG Wen-zhi, YANG Ben-peng, CAI Wen-wei, XIONG Guo-ru, WANG Jun-gang, FENG Cui-lian, ZHANG Shu-zhen   

  1. Sugarcane Research Center of Chinese Academy of Tropical Agricultural Sciences, Institute of Tropical Bioscience and Biotechnology of Chinese Academy of Tropical Agricultural Sciences, Key Laboratory of Biology and Genetic Resources of Tropical Crops of Ministry of Agriculture, Haikou 571101
  • Received:2015-03-27 Published:2016-01-09 Online:2016-01-22

摘要: 通过多基因遗传转化策略, 水稻、玉米等农作物实现了一次多个性状的遗传改良, 为探索甘蔗一次多基因遗传转化的可行性, 用一个多基因植物表达载体, 对甘蔗进行遗传转化。通过多重PCR法对4个外源基因在各个转基因株系内的整合与缺失进行检测。结果证明, 甘蔗一次多基因遗传转化是可行的, 但因插入的DNA片段较大, 会发生基因丢失现象。因此, 一次多基因遗传转化需要在转化的过程中获得相对较多的转化株系, 才能从中筛选到有应用价值的株系。

关键词: 甘蔗, 农杆菌介导法, 多基因遗传转化, 多重PCR检测

Abstract: Multiple traits of crops such as rice and corn had been improved in single time of transformation by the strategy of multiple gene transformation. For exploring the feasibility of multiple gene transformation of sugarcane in this research, a multiple gene expression vector was used and transformed to sugarcane and many transformed plants were produced. Multiple PCR method was used to analyze the integration and deficiency of multiple genes in the transformed plants. Results showed that multiple gene transformation of sugarcane was feasible. But due to the insertion of large DNA fragments some genes were lost during the integration. Thus it is a need to have sufficient transformed plants then valuable transformed plants can be selected for further research.

Key words: sugarcane, Agrobacterium-mediated, multiple gene transformation, multiple PCR detection