遗传 ›› 2010, Vol. 32 ›› Issue (5): 492-497.

• 研究报告 • 上一篇    下一篇

大豆醛脱氢酶基因GmALDH3-1的克隆及在生殖器官中的表达

  

  1. 南京农业大学国家大豆改良中心 作物遗传与种质创新国家重点实验室, 南京 210095  

  • 收稿日期:2009-10-10 修回日期:2009-11-12 出版日期:2010-05-20 发布日期:2010-05-05
  • 通讯作者: 黄方 E-mail:fhuang@njau.edu.cn
  • 基金资助:

    国家自然科学基金;国家自然科学基金;教育部博士点基金;江苏省自然科学基金;南京农业大学青年创新基金

Cloning and characterization of an aldehyde dehydrogenase gene GmALDH3-1in soybean

HUANG Fang, CHI Ying-Jun, HE Hui, YU De-Yue
  

  1. National Center for Soybean Improvement, National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China

  • Received:2009-10-10 Revised:2009-11-12 Online:2010-05-20 Published:2010-05-05

摘要:

通过基因芯片技术,从大豆中鉴定了一个花优势表达基因,其在大豆花中的表达量为叶片中的85倍。通过生物信息学方法,拼接了该基因的全长序列,并通过RT-PCR克隆了该基因。BLAST检索分析表明该基因编码醛脱氢酶,命名为GmALDH3-1。GmALDH3-1包含一个1485 bp的开放阅读框,编码494个氨基酸残基。GmADLH3-1与白杨的醛脱氢酶PtALDH3相似性最高(氨基酸相似率83%,一致率为68%),而与来自于人的ALDH3B的氨基酸一致率和相似率分别为39%和59%。系统发生分析表明GmALDH3-1与其它植物ALDH3亚家族成员位于一个分支,且与白杨PtALDH3和拟南芥AtALDH3F1亲缘关系最近。采用实时定量RT-PCR检测了GmALDH3-1基因在大豆叶、根和花中的表达,结果表明GmALDH3-1基因在花中高丰度表达,在根和叶中未检测到表达。运用基因芯片信息分析了GmALDH3-1在种子发育过程中的表达情况,结果表明GmALDH3-1在种子发育过程中的外表皮、内表皮、外胚珠和种脐中表达量较高。文章讨论了GmALDH3-1基因在大豆生殖器官发育中可能发挥的作用。

关键词: 大豆, 醛脱氢酶, 生殖器官, 基因表达

Abstract:

A gene predominately expressed in soybean flowers was initially identified by microarray analysis. Through bioinformatics approach and RT-PCR, the full-length gene was cloned from soybean flowers. The BLAST searches indicated this gene encoded an aldehyde dehydrogenase and named as GmALDH3-1. GmALDH3-1 contains a complete open reading frame of 1485 bp which encodes a peptide of 494 amino acids. The GmALDH3-1encoding product shows 83% and 68% sequence similarity and identity with Populus tomentosa PtALDH3 and even shows 39% and 59% with human ALDH3B. The phylogenetic analysis shows that GmALDH3-1 and other ALDH2 subfamily members group into a branch and GmALDH3-1 is closest to PtALDH3 and Arabidopsis AtALDH3F1. By real-time RT-PCR, it was found that expression of GmALDH3-1 was highest in flowers, almost undetectable in leaf and root. Further we analyzed GmALDH3-1 expression during the course of seed development based on publicly available microarray data and found that GmALDH3-1 was highly expressed in the seed endothelium, epidermis, outer integument and hilum. The putative functions for GmALDH3-1 in soybean reproductive development are discussed.

Key words: soybean, aldehyde dehydrogenase, reproductive tissues, gene expression