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作物学报 ›› 2011, Vol. 37 ›› Issue (01): 40-47.doi: 10.3724/SP.J.1006.2011.00040

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

两个棉花ß-葡聚糖酶新基因的结构与表达特征分析

 董佳, 蔡彩平, 王立科, 赵亮, 张天真, 郭旺珍*   

  1. 南京农业大学 / 作物遗传与种质创新国家重点实验室, 江苏南京 210095
  • 收稿日期:2010-06-10 修回日期:2010-08-04 出版日期:2011-01-12 网络出版日期:2010-11-12
  • 通讯作者: 郭旺珍, E-mail: moelab@njau.edu.cn
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2006AA10Z111)，转基因生物新品种培育重大专项(2008ZX08009-003)和教育部111项目(B08025)资助。

Structure and Expression Analysis of Two Novel Genes Encoding β-glucanase in Cotton

DONG Jia,CAI Cai-Ping,WANG Li-Ke,ZHAO Liang,ZHANG Tian-Zhen,GUO Wang-Zhen*   

  1. National Key Laboratory of Crop Genetics & Germplasm Enhancement / Nanjing Agricultural University, Nanjing 210095, China
  • Received:2010-06-10 Revised:2010-08-04 Published:2011-01-12 Published online:2010-11-12
  • Contact: 郭旺珍, E-mail: moelab@njau.edu.cn

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摘要： β-葡聚糖酶是一类能降解β-葡聚糖的水解酶。本研究分别通过电子克隆和棉纤维发育cDNA文库筛选法克隆到2个棉花β-葡聚糖酶新基因，内切-1,4-β-葡聚糖酶基因(GhEG，GenBank登录号为HM462003)和1,3-β-葡聚糖酶基因(GhGLU，GenBank登录号: HM462004)。GhEG全长ORF为1 581 bp，编码526个氨基酸残基。GhGLU全长ORF为1 410 bp，编码469个氨基酸残基。基因组水平分析表明，GhEG含5个内含子和6个外显子，而GhGLU无内含子，仅1个外显子。新克隆的2个基因在二倍体棉种非洲棉和雷蒙德氏棉中含1个拷贝，而在四倍体陆地棉和海岛棉中存在2个拷贝。通过开发SNP标记分别将GhEG和GhGLU在四倍体中的一个拷贝定位在第19染色体和第4染色体上。Q-PCR表达分析表明，GhEG在根、茎、叶中表达水平很低，而在纤维伸长期优势表达，在15 DPA和20 DPA纤维中，该基因在海岛棉海7124中的转录本显著高于陆地棉TM-1。GhGLU在根、茎、叶及纤维发育不同时期均有表达，属于组成性表达基因，特别在根、纤维发育初始期和伸长后期优势表达，且表达水平在陆地棉TM-1和海岛棉海7124间也有显著差异。

关键词: 内切-1,4-β-葡聚糖酶, 1,3-β-葡聚糖酶, 结构, 表达

Abstract: The β-glucanase is a type of enzyme degrading β-glucan. Cloning and expression analysis of genes encoding β-glucanase can provide information in both gene resources and breeding utilization for improvement of cotton fiber quality. The two novel genes encoding β-glucanase, designated as GhEG (GenBank accession No: HM462003) and GhGLU (GenBank accession No: HM462004), were obtained by sillico cloning and cDNA library screening, respectively. GhEG contained an open reading frame of 1 581 bp that encoded a polypeptide of 526 amino acids, and GhGLU contained an open reading frame of 1 410 bp that encoded a polypeptide of 469 amino acids. The genome sequence indicated that GhEG has five introns and six exons, while GhGLU has no intron and only one exon. The two genes all had one copy in diploid cotton species G. herbaceum and G. raimondii and two copies in tetraploid cotton species G. hirsutum acc. TM-1 and G. barbadense cv. Hai 7124, respectively. One of GhEG or GhGLU homoelogs in tetraploid was located on chromosome 19 and chromosome 4 by developing SNP marker, respectively. Q-PCR expression analysis showed that GhEG was expressed specifically in fiber elongation and had obvious difference between TM-1 and Hai7124 at the fiber elongation period of 15 DPA and 20 DPA, almost no transcripts were detected in root, stem and leaf. GhGLU was expressed in all tissues, dominantly in root, at fiber initiation and fiber late elongation phase, with obvious difference between TM-1 and Hai7124.

Key words: GhEG, GhGLU, Structure, Expression

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