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园艺学报 ›› 2010, Vol. 37 ›› Issue (6): 939-948.

• 观赏植物 • 上一篇    下一篇

苹果属观赏海棠McANS基因克隆与不同叶色品种间表达差异分析

田 佶,沈红香,张 杰,姚允聪*,宋婷婷,耿 慧

  

  1. (北京农学院植物科学技术系,北京 102206)
  • 收稿日期:2009-11-02 修回日期:2010-04-29 出版日期:2010-06-25 发布日期:2010-06-25
  • 通讯作者: 姚允聪

Cloning of McANS Gene in Malus Crabapple and Expression Analysis in Different Cultivars

TIAN Ji,SHEN Hong-xiang,ZHANG Jie,YAO Yun-cong*,SONG Ting-ting,and GENG Hui*   

  1. (Department of Plant Science and Technology,Beijing University of Agriculture,Beijing 102206,China)
  • Received:2009-11-02 Revised:2010-04-29 Online:2010-06-25 Published:2010-06-25
  • Contact: YAO Yun-cong

摘要: 以苹果属观赏海棠品种‘王族’叶片总RNA为模板,通过RT-PCR与RACE扩增,获得一个1 350 bp的花色素花青苷元合成酶(anthocyanidin synthase,ANS)基因的cDNA序列,其基因编码区共1 074 bp,编码357个氨基酸,命名为McANS(登录号FJ817488)。基因组序列全长1 268 bp,有1个内含子,内含子的剪切位点均符合真核生物“GT-AG”规则。使用实时荧光定量PCR和紫外可见光分光光度计法,对3种不同叶色的观赏海棠品种‘火焰’(叶片绿色)、‘绚丽’(新叶红色)和‘王族’(叶片紫色)幼叶和功能叶中的McANS表达量、花青苷和类黄酮含量进行测定分析,结果表明:McANS在3个品种的幼叶和功能叶中均有表达,在幼叶中表达量显著高于功能叶,在‘绚丽’中表达量相差达到23.82倍。花青苷含量的变化与McANS相对表达量变化趋势具有一致性,而类黄酮含量的变化与McANS相对表达量无明显相关。说明McANS在苹果属观赏海棠叶片花青苷代谢及色泽形成过程中具有重要作用。

关键词: 苹果属, 观赏海棠, 花青苷元合成酶, 花青苷, 类黄酮, 实时荧光定量PCR

Abstract: Using the total RNA from the leaves of Malus crabapple‘Royalty’as the template,the full cDNA of ANS(anthocyanidin synthase)gene(1 350 bp)was cloned by reverse transcription polymerase chain reaction(RT-PCR)and rapid-amplification of cDNA ends(RACE). The gene was named as McANS,containing an open reading frame(1 074 bp)and encoding a protein of 357 amino acids. Corresponding DNA sequence is 1 268 bp,containing one intron,and all the cleave sites obey with the GT-AG rule. The expression of McANS and the content of anthocyanins and flavonoids was determined by real-time quantitative PCR and spectrophotometer respectively in the mature and young leaves of M.‘Flame’(green young and mature leaf),M.‘Radiant’(red young leaf and green mature leaf),M.‘Royalty’(purple young and mature leaf). The results showed that McANS was expressed in both mature leaves and young leaves of the above three cultivars,and the expression level of McANS in young leaves is higher than in mature leaves,23.82-fold difference in M. ‘Radiant’. The trends of anthocyanins content had significant consistency with that of the relative expression of McANS gene. But flavonoids content had not relevance with relative expression of McANS gene. It’s description that McANS have very close relationship with the metabolism of anthocyanins and color of leaf.

Key words: Malus, ornamental crabapples, ANS, anthocyanin, flavonoid, real-time quantitative PCR

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