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作物学报 ›› 2012, Vol. 38 ›› Issue (09): 1649-1656.doi: 10.3724/SP.J.1006.2012.01649

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

棉花S-腺苷甲硫氨酸脱羧酶基因的克隆及低温下的表达分析

耿卫东，李艳军，张新宇，朱华国*，孙杰*   

  1. 石河子大学农学院 / 新疆生产建设兵团绿洲生态农业重点实验室，新疆石河子832003
  • 收稿日期:2012-01-08 修回日期:2012-04-20 出版日期:2012-09-12 网络出版日期:2012-06-04
  • 通讯作者: 朱华国, Email: zhgroger@sohu.com; 孙杰, E-mail: sunjiezh@yahoo.com.cn
  • 基金资助:

    本研究由兵团博士资金项目(2011BB004)和国家转基因生物新品种培育重大专项子课题(2008ZX08005-005)资助。

Molecular Cloning and Expression Analysis of GhSAMDC at Low Temperature Stress in Cotton (Gossypium hirsutum L.)

GENG Wei-Dong,LI Yan-Jun,ZHANG Xin-Yu,ZHU Hua-Guo*,SUN Jie*   

  1. College of Agriculture / The key Laboratory of Oasis Eco-agriculture Xinjiang Production and Construction Group, Shihezi University, Shihezi 832003, China
  • Received:2012-01-08 Revised:2012-04-20 Published:2012-09-12 Published online:2012-06-04
  • Contact: 朱华国, Email: zhgroger@sohu.com; 孙杰, E-mail: sunjiezh@yahoo.com.cn

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被引次数

12

摘要： 根据棉花S-腺苷甲硫氨酸脱羧酶(S-adenosylmethionine decarboxylase, SAMDC)基因已知EST序列设计引物，采用RACE和RT-PCR技术克隆获得该基因的全长cDNA序列，命名为GhSAMDC (GenBank登录号为JN020148)。生物信息学分析表明，GhSAMDCcDNA序列全长1 874 bp，涵盖tiny ORF (tORF)、upstream ORF (uORF)和main ORF (mORF) 3个植物SAMDC基因特征ORF。其中mORF长1 064 bp，编码含355个氨基酸残基的SAMDC酶原，预测分子量为38.25 kD，该酶原含有高度保守的酶原剪切位点结构域(LSESSLF)和与SAMDC蛋白快速降解有关的PEST (TIHVTPEDGFSYAS)结构域。GhSAMDC基因组DNA序列全长2 743 bp，包含3个内含子，均位于5' UTR，其中一个位于uORF内。聚类分析表明，GhSAMDC与葡萄中该蛋白的同源关系最近，并且与其他双子叶植物聚为一类。荧光定量PCR分析结果表明，GhSAMDC表达受低温诱导，在冷敏感品种新陆早1号中基因表达水平明显高于在耐冷品种新陆早33号中。

关键词: 棉花, S-腺苷甲硫氨酸脱羧酶, 序列分析, 定量PCR, 低温表达

Abstract: S-adenosylmethionine decarboxylase is one of the key enzymes in polyamine biosynthesis. With the increase of polyamine accumulation in plants, SAMDC can improve crop tolerance to environment stresses. We cloned the S-adenosylmethionine decarboxylase gene from Gossypium hirsutum by RACE and RT-PCR, named GhSAMDC (GenBank accession number: JN020148). The cDNA full length was 1 874 bp, with three open reading frames, i.e. tiny ORF (tORF), upstream ORF (uORF) and main ORF (mORF). The mORF was 1 064 bp encoding 355 amino acids with a molecular mass of 38.25 kD, and its deduced protein had two highly conserved function domains (proenzyme cleavage site and PEST domain). The complete genomic DNA sequence of GhSAMDC was 2 743 bp in length with three introns, all of which were located in 5' UTR, and one lay in theuORF. Phylogenetic analysis showed GhSAMDC was the closest to that in Vitis vinifera, and Real-time PCR suggested that GhSAMDC was induced by cold stress. It showed lower gene expression level in cold resistant cultivar Xinluzao 33 than in cold sensitive cultivar Xinluzao 1.

Key words: Cotton (Gossypium hirsutum L.), S-adenosylmethionine decarboxylase, Sequence analysis, qPCR, Expression at low temperature

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