中国农业科学 ›› 2007, Vol. 40 ›› Issue (12): 2688-2695 .

• 作物遗传育种·种质资源 • 上一篇    下一篇

芥菜型油菜类黄酮合成相关基因的克隆和序列分析

严明理,刘忠松,官春云,陈社员,刘显军,袁谋志   

  1. 湖南农业大学油料作物研究所
  • 收稿日期:2006-11-06 修回日期:1900-01-01 出版日期:2007-12-10 发布日期:2007-12-10
  • 通讯作者: 刘忠松

Cloning and Sequence Analysis of Genes for Flavonoid Biosynthesis in Brassica juncea

  

  1. 湖南农业大学油料作物研究所
  • Received:2006-11-06 Revised:1900-01-01 Online:2007-12-10 Published:2007-12-10

摘要: 【目的】分离和克隆芥菜型油菜类黄酮合成相关基因。【方法】采用同源克隆基因的方法,参照拟南芥等植物控制类黄酮合成的基因保守序列设计引物,对扩增片段进行测序并进行BLAST分析。【结果】有13对引物扩增的17个基因拷贝与参考基因序列相符,这些克隆属于13个已知功能基因,其中查尔酮合成酶基因有3个不同的基因拷贝,花色素形成(Production of anthocyanin pigment)基因和查尔酮异构酶基因分别有2个不同的基因拷贝,其余引物的扩增只获得一个拷贝。在GenBank数据库中进行检索表明,所克隆的基因拷贝中的DNA结合/转录因子基因(TT2、TT8、TTG2) 、花色素形成基因(PAP)和黄烷酮-3-羟化酶基因(TT6),在芸薹属植物中未见报道。【结论】克隆芥菜型油菜类黄酮合成相关基因拷贝为阐明油菜种皮颜色形成的遗传调节奠定了基础。

关键词: 芥菜型油菜, 类黄酮, 基因

Abstract: 【Objective】 The genes for flavonoid biosynthesis are to be cloned in Brassica juncea .【Method】Homology-based cloning strategy was used in this study. The primers were designed according to the sequences of homologous genes of the model plant Arabidopsis thaliana. The gene fragments were sequenced, and analyzed by BLAST.【Result】The 17 fragments were cloned and sequenced from 13 pairs of specific primers, which were confirmed to correspond to the Arabidopsis thaliana homologous genes and belong to the 13 known functional genes. The gene chs for chalcone synthase has 3 fragments different in size, while the genes pap for production of anthocyanin pigment and chi for chalcone isomerase have 2 fragments each. 【Conclusion】The homology- based cloning of the genes for flavonoid biosynthesis in Brassica juncea is viable. The cloned fragments of genes for flavonoid biosynthesis will provide the foundations for uncovering the genetic regulation of seed coat color formation in Brassica juncea.

Key words: Brassica juncea, Flavonoid, Genes, Homology-based cloning