Journal of Nuclear Agricultural Sciences ›› 2015, Vol. 29 ›› Issue (4): 692-698.DOI: 10.11869/j.issn.100-8551.2015.04.0692

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The Maize Transcription Factor ZmPTF1 Expression in Transgenic Tobacco

LIU Chunhua, GAO Jianrui, WANG Peng, XIA Chao, DONG Xiaoqiu, LIU Bing, ZHAO Dan, WEI Rujun, LAN Hai   

  1. Maize Research Institute, Sichuan Agricultural University, Chengdu, Sichuan 611134
  • Received:2014-03-14 Revised:2014-11-10 Online:2015-04-13 Published:2015-04-16

玉米转录因子基因ZmPTF1在转基因烟草中的表达

刘春华, 高剑瑞, 王鹏, 夏超, 董晓秋, 刘兵, 赵丹, 韦如俊, 兰海   

  1. 四川农业大学玉米所, 四川 成都 611134
  • 通讯作者: 兰海, 男, 副研究员, 主要从事作物遗传育种研究.E-mail:19110589@qq.com
  • 作者简介:刘春华, 女, 主要从事玉米遗传育种研究.E-mail:584423632@qq.com
  • 基金资助:

    国家“863”计划(2011AA10A103)

Abstract:

To investigate the effect of the transcription factor ZmPTF1 in the germination of corn seeds, primers were designed according to the maize PTF1 sequence recorded in GeneBank. Full-length cDNA sequence. of ZmPTF1 was cloned from normal maize inbred line A318, with a open reading frame consisting of 1 446bp bases and 481amino acids. Construct plant expression vector PCAMBIA3301-ZmPTF1 and transform into tobacco W38. Positive transgenic tobaccos were identified using antibiotic screening test and PCR detection. Using southern hybridization and PCR detection to analyze the normally grown T1 transgenic tobaccos, and the result showed that the exogenous ZmPTF1 was successfully integrated into transgenic plant genome and could be transcribed and inherited normally. The germination rate and the α-amylase activity of the transgenic tobacco seeds increased significantly, indicating that the overexpression of ZmPTF1 gene promoted the tobacco seeds break dormancy in advance and enter the stage of germination and therefore increased thegermination potential and germination rate. The results will provide further theoretical basis for the research of functions of ZmPTF1.

Key words: ZmPTF1, tobacco, germination, α-amylase, southern hybridization

摘要:

为了探讨转录因子基因ZmPTF1在玉米种子萌发中的作用, 本研究根据GenBank中收录的玉米PTF1序列设计PCR引物, 利用普通玉米自交系A318克隆得到ZmPTF1的全长cDNA序列.该序列开放阅读框包括1 446bp碱基, 由481个氨基酸组成.本文构建植物表达载体PCAMBIA3301-ZmPTF1 转化烟草W38, 经过抗生素筛选和PCR鉴定得到转基因阳性烟草, 对2株生长正常的T1转基因烟草进行southern杂交分析及RT-PCR 检测, 结果表明外源ZmPTF1已经整合进了烟草基因组中并且能正常转录并遗传.与对照相比, 转基因烟草种子的发芽率、发芽势及萌发种子α-淀粉酶活性显着增高, 表明ZmPTF1基因过表达会促进烟草种子提前解除休眠进入发芽阶段, 提高烟草种子发芽势和发芽率.该研究结果为进一步研究ZmPTF1基因的功能提供理论依据.

关键词: ZmPTF1, 烟草, 萌发, α-淀粉酶, southern杂交