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作物学报 ›› 2015, Vol. 41 ›› Issue (03): 479-486.doi: 10.3724/SP.J.1006.2015.00479

• 研究简报 • 上一篇    下一篇

飞机草类黄酮3’-羟化酶基因(CoF3’H)的克隆及其在烟草中的表达

何海旺,潘华清,张铙丹,何龙飞*   

  1. 广西大学农学院,广西南宁 530004
  • 收稿日期:2014-08-12 修回日期:2014-12-19 出版日期:2015-03-12 网络出版日期:2014-12-29
  • 通讯作者: 何龙飞, E-mail: lfhe@gxu.edu.cn, Tel: 0771-3235212-801
  • 基金资助:

    本研究由广西自然科学基金项目(桂科自0832047)资助。

Chromolaena odorata Flavonoid 3’-hydroxylase Gene Cloning and Its Expression in Tobacco

HE Hai-Wang,PAN Hua-Qing,ZHANG Nao-Dan,HE Long-Fei*   

  1. Agricultural College of Guangxi University, Nanning 530004, China
  • Received:2014-08-12 Revised:2014-12-19 Published:2015-03-12 Published online:2014-12-29
  • Contact: 何龙飞, E-mail: lfhe@gxu.edu.cn, Tel: 0771-3235212-801

摘要:

利用RT-PCR和RACE技术从飞机草中克隆得到1628 bp的类黄酮3’-羟化酶cDNA的全长序列,命名为CoF3H,GenBank登录号为HQ268505.1。CoF3’H基因的编码区长度为1524 bp,编码507个氨基酸。氨基酸序列含P450蛋白结构域和半胱氨酸亚铁血红素结合域保守区(F××G×R×C×G),利用DNAMAN软件比对显示,CoF3’H与其他植物的F3’H蛋白的同源性很高。通过农杆菌介导成功地将CoF3H基因导入烟草,T2代的转CoF3H基因烟草植株的黄酮含量显著高于野生型。说明CoF3¢H在黄酮的生物合成中起重要作用,为后续研究飞机草的化感作用及综合利用提供基础。

关键词: 飞机草, 类黄酮3’-羟化酶, 克隆, 表达

Abstract:

A flavonoid 3’-hydroxylase gene cDNA sequence was cloned by RACE and RT-PCR techniques from Chromolaena odorata. The obtained full-length cDNA was named as CoF3H with GenBank accession number HQ268505.1. It is 1628 bp in length, containing a 1524 bp open reading frame, encoding 507 amino acid residues. The amino acid sequence of CoF3H contains cytochrome P450 domain and cysteineheme binding region (F××G×R×C×G). Homology analysis by DNAMAN software showed that the deduced CoF3H protein was highly homologous to F3’H proteins from different species. CoF3H gene was transferred into tobacco by agrobacterium-mediated genetic transformation. There was higher content of flavonvoids in T2 transgenic tobacco than in the wild type. This result shows that CoF3H plays an important role in flavonoid biosynthesis, and provides a basis for researches on allelopathy and comprehensive utilization of C. odorata in the future.

Key words: Chromolaena odorata, Flavonoid 3’-hydroxylase, Clone, Expression

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