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园艺学报 ›› 2010, Vol. 37 ›› Issue (2): 269-276.

• 观赏植物 • 上一篇    下一篇

苹果属观赏海棠McCHS 基因的克隆及实时定量表达

宋婷婷;沈红香;姚允聪*;田佶
  

  1. (北京农学院植物科学技术系,北京102206)
  • 收稿日期:2009-04-10 修回日期:2010-01-08 出版日期:2010-02-25 发布日期:2010-02-25
  • 通讯作者: 姚允聪

Studies on Cloning and Real-time Expression of McCHS Gene in MalusCrabapple

SONG Ting-ting;SHEN Hong-xiang;YAO Yun-cong;TIAN Ji   

  1. (Department of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206, China)
  • Received:2009-04-10 Revised:2010-01-08 Online:2010-02-25 Published:2010-02-25
  • Contact: YAO Yun-cong

摘要: 以苹果属观赏海棠‘王族’品种叶片提取的总RNA 为模板,通过RT-PCR 与RACE 扩增,获
得一个1 529 bp 的查尔酮合成酶(Chalcone sythase,CHS)基因的cDNA 序列,其基因编码区共1 167 bp,编码389 个氨基酸,命名为McCHS。使用实时荧光定量PCR 和紫外可见光分光光度计,对3 类不同叶色的观赏海棠品种‘火焰’(Malus‘Flame’,叶片绿色)、 ‘绚丽’(Malus‘Radiant’,新叶红色)、‘高原之火’(Malus‘Prairifire’,新叶红色)和王族(Malus‘Royalty’,叶片紫色)幼叶和功能叶中的McCHS 表达及其花色苷含量进行测定分析,结果表明:McCHS 在4 个品种的幼叶和功能叶中均有表达, 其幼叶表达量的变化与叶片中花色苷含量的变化一致;除‘绚丽’外的功能叶表达量变化和花色苷含量变化也一致,‘绚丽’功能叶的特殊情况可能涉及花色苷合成途径中的其他酶和转录因子。

关键词: 苹果属, 观赏海棠, 查尔酮合成酶基因, 花色苷, 荧光定量

Abstract: Using the total RNA from the leaves of Malus crabapple‘Royalty’as the template, the full cDNA of CHS(Chalcone synthase)gene(1 529 bp)was cloned by reverse transcription polymerse chain reaction(RT-PCR)and rapid-amplification of cDNA ends(RACE). The gene was named McCHS, containing
an open reading frame(1 167 bp)and encoding a protein of 389 amino acids. The expression of McCHS and the content of anthocyanin was determined by Real-time quantitative PCR and spectrophotometer
respectively in the mature and young leaves of ‘Flame’(green young and mature leaf),‘Radiant’(red young leaf and green mature leaf),‘Prairifire’(red young leaf and green mature leaf),‘Royalty’(purple
young and mature leaf). The results showed that McCHS was expressed in both mature leaves and young leaves of the above four cultivars. It was in the young leaves of all the four cultivars that the variation in the
McCHS expression was similar to that in the anthocyanin content. The same trend was found in the mature leaves of most cultivars with the exception of ‘Radiant’, which may have other enzymes and transcriptionfactors involved in the anthocyanin synthesis in its mature leaves.

Key words: Malus, crabapples, chalcone synthase, anthocyanin, real-time quantitative PCR