遗传 ›› 2014, Vol. 36 ›› Issue (7): 707-712.doi: 10.3724/SP.J.1005.2014.0707

• 研究报告 • 上一篇    下一篇

大豆查尔酮还原酶基因CHR1的功能研究

吴楠1, 王丕武1, 李丹1, 代力强1, 郑成忠1, 卢实1, 才源1, 张卓1, 曲静1, 夏海丰1, 2   

  1. 1. 吉林农业大学农学院, 长春 130118;
    2. 通化市农业科学研究院, 梅河口 135000
  • 收稿日期:2014-01-07 出版日期:2014-07-20 发布日期:2014-06-23
  • 通讯作者: 王丕武, 教授, 博士生导师, 研究方向:生物技术与作物遗传育种。E-mail: peiwuw@163.com
  • 作者简介:吴楠, 在读硕士研究生, 专业方向:生物技术与作物遗传育种。E-mail:525762426@qq.com
  • 基金资助:
    国家自然科学基金项目(编号:31171568)资助

Function of chalcone reductase gene CHR1 in soybean

Nan Wu1, Piwu Wang1, Dan Li1, Liqiang Dai1, Chengzhong Zheng1, Shi Lu1, Yuan Cai1, Zhuo Zhang1, Jing Qu1, Haifeng Xia1, 2   

  1. 1. College of Agriculture, Jilin Agricultural University, Changchun 130118, China;
    2. Tonghua Academy of Agricultural Sciences, Meihekou, Jilin 135000, China
  • Received:2014-01-07 Online:2014-07-20 Published:2014-06-23

摘要: 为了验证查尔酮还原酶基因CHR1在大豆苷元合成中的作用, 文章克隆了CHR1基因并构建了RNA干扰表达载体pCPB-CHR1-RNAi, 将载体转化受体大豆品种“吉农28”中, 以期抑制CHR1基因的转录。通过农杆菌介导的遗传转化和PCR检测得到4株T0 代阳性植株, 13株T1代阳性植株。Southern blotting结果表明, 功能元件以单拷贝的形式整合到大豆的基因组中。利用实时荧光定量PCR法(Quantitative real-time PCR, qRT-PCR)测定CHR1基因在mRNA水平上的表达量, 结果表明, 转基因大豆植株中CHR1的表达量与未转化的受体大豆相比降低了60%~99%; 高效液相色谱法(High performance liquid chromatography, HPLC)检测到合成大豆苷元过程中的前体物质异甘草素的含量降低了38.7%。该RNA干扰机制在转录水平上抑制了CHR1基因的表达。

关键词: 大豆, 查尔酮还原酶CHR1, 大豆苷元, RNA干扰载体

Abstract: To verify the function of chalcone reductase gene (CHR1) in soybean Daidzein synthesis, CHR1 gene in soybean was cloned, and an RNAi expression vector pCPB-CHR1-RNAi was constructed. Four transformed plants in T0 generation and thirteen transformed plants in T1 generation of soybean “Jinong28” were obtained by agrobacterium-mediated genetic transformation, in which transcriptions of CHR1 gene were depressed. Southern blotting showed the functional fragment of pCPB-CHR1-RNAi was integrated into the genome of recipient soybean in the form of a single copy. Detection of the transcription of CHR1 gene using quantitative real-time PCR (qRT-PCR) showed that the expression of CHR1 gene in transformed plants decreased 60%-99% compared to the recipient soybean, while the content of isoliquiritigenin, the precursors of daidzein, decreased 38.7%. These results indicate that RNA interference can suppress the transcription of CHR1 gene expression successfully.

Key words: soybean, chalcone reductase CHR1, daidzein, RNA interference vector