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丹参肉桂醇脱氢酶基因(SmCAD)的克隆及表达分析

葛茜, 邝静, 武玉翠, 张媛, 肖娅萍, 王喆之

葛茜, 邝静, 武玉翠, 张媛, 肖娅萍, 王喆之. 丹参肉桂醇脱氢酶基因(SmCAD)的克隆及表达分析[J]. 植物科学学报, 2013, 31(3): 261-268. DOI: 10.3724/SP.J.1142.2013.30261
引用本文: 葛茜, 邝静, 武玉翠, 张媛, 肖娅萍, 王喆之. 丹参肉桂醇脱氢酶基因(SmCAD)的克隆及表达分析[J]. 植物科学学报, 2013, 31(3): 261-268. DOI: 10.3724/SP.J.1142.2013.30261
GE Qian, KUANG Jing, WU Yu-Cui, ZHANG Yuan, XIAO Ya-Ping, WANG Zhe-Zhi. Cloning and Expression Analysis of Cinnamyl Alcohol Dehydrogenase (SmCAD) Gene in Salvia miltiorrhiza Bunge[J]. Plant Science Journal, 2013, 31(3): 261-268. DOI: 10.3724/SP.J.1142.2013.30261
Citation: GE Qian, KUANG Jing, WU Yu-Cui, ZHANG Yuan, XIAO Ya-Ping, WANG Zhe-Zhi. Cloning and Expression Analysis of Cinnamyl Alcohol Dehydrogenase (SmCAD) Gene in Salvia miltiorrhiza Bunge[J]. Plant Science Journal, 2013, 31(3): 261-268. DOI: 10.3724/SP.J.1142.2013.30261
葛茜, 邝静, 武玉翠, 张媛, 肖娅萍, 王喆之. 丹参肉桂醇脱氢酶基因(SmCAD)的克隆及表达分析[J]. 植物科学学报, 2013, 31(3): 261-268. CSTR: 32231.14.SP.J.1142.2013.30261
引用本文: 葛茜, 邝静, 武玉翠, 张媛, 肖娅萍, 王喆之. 丹参肉桂醇脱氢酶基因(SmCAD)的克隆及表达分析[J]. 植物科学学报, 2013, 31(3): 261-268. CSTR: 32231.14.SP.J.1142.2013.30261
GE Qian, KUANG Jing, WU Yu-Cui, ZHANG Yuan, XIAO Ya-Ping, WANG Zhe-Zhi. Cloning and Expression Analysis of Cinnamyl Alcohol Dehydrogenase (SmCAD) Gene in Salvia miltiorrhiza Bunge[J]. Plant Science Journal, 2013, 31(3): 261-268. CSTR: 32231.14.SP.J.1142.2013.30261
Citation: GE Qian, KUANG Jing, WU Yu-Cui, ZHANG Yuan, XIAO Ya-Ping, WANG Zhe-Zhi. Cloning and Expression Analysis of Cinnamyl Alcohol Dehydrogenase (SmCAD) Gene in Salvia miltiorrhiza Bunge[J]. Plant Science Journal, 2013, 31(3): 261-268. CSTR: 32231.14.SP.J.1142.2013.30261

丹参肉桂醇脱氢酶基因(SmCAD)的克隆及表达分析

基金项目: 陕西省自然科学基础研究计划项目(2012JQ4013);中央高校基本科研业务费专项资助资金(GK201102017);陕西省教育厅科学技术研究发展计划(12JK0833)。
详细信息
    作者简介:

    葛茜(1988- ),女,硕士,研究方向为植物分子生物学、植物基因工程(E-mail: geqian@stu.snnu.edu.cn)。

    通讯作者:

    王喆之,教授,博士生导师(E-mail: zzwang@snnu.edu.cn)。

  • 中图分类号: Q943.2

Cloning and Expression Analysis of Cinnamyl Alcohol Dehydrogenase (SmCAD) Gene in Salvia miltiorrhiza Bunge

  • 摘要: 肉桂醇脱氢酶(cinnamyl alcohol dehydrogenase, CAD)依赖于NADPH还原肉桂醛及其衍生物,是催化木质素单体生物合成途径的最后一步关键酶。通过分析丹参转录组数据库,从丹参中获得一条肉桂醇脱氢酶基因,命名为SmCAD (Genebank注册号:HQ162287)。该序列包含一个长为1083 bp的开放阅读框,有3个内含子和4个外显子,编码360个氨基酸,含有NADP(H)结合域,Zn1和Zn2锌结合位点。利用BD walking的方法获得其启动子序列1202 bp,序列分析结果表明,SmCAD启动子区包含茉莉酸甲酯(MeJA)、脱落酸(ABA)、赤霉素(GA3)响应元件以及MYB结合位点。利用实时荧光定量PCR分析表明,该基因在丹参根、茎、叶中均有表达,且其表达受到MeJA的诱导和GA3的抑制,推测该基因可能参与了丹参对外源信号的应答反应。研究结果可为进一步研究SmCAD基因在丹参中的具体功能提供理论依据。
    Abstract: Cinnamyl alcohol dehydrogenase (CAD) catalyzes the reduction reaction of cinnamic aldehyde and its derivatives depending on the NADPH.It is the key enzyme in the last step of lignin biosynthesis. A novel CAD gene designated as SmCAD (GenBank accession No.: HQ162287) was cloned from Salvia miltiorrhiza Bunge according to the transcription databases. It contained a 1083 bp open reading frame, including 3 introns and 4 exons, and encoded a protein of 360 amino acids. The predicted SmCAD protein had Zn1 and Zn2 binding sites and NADP(H) binding domain. A total of 1202 bp promoter sequences were obtained using BD walking method, and sequence analysis revealed that there were MeJA, ABA, GA3 response elements and MYB binding sites. Real-time quantitative PCR showed that SmCAD was expressed in different organs, and its expression was induced by MeJA and inhibited by GA3. These results suggested that this gene may be involved in the response of exogenous signals. This study contributes to the theoretical basis of the specific function of SmCAD.
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出版历程
  • 收稿日期:  2012-10-22
  • 修回日期:  2012-12-18
  • 发布日期:  2013-06-29

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