作物学报Issue(2)：222-229,8.DOI:10.3724/SP.J.1006.2013.00222
三个陆地棉水孔蛋白基因的克隆与表达分析
Cloning and Expression Analysis of Three Aquaporin Genes in Upland Cotton (Gossypium hirsutum L.)
摘要
Abstract
To provide an insight into molecular mechanism of cotton fiber development, we analyzed the sequence characteristics of three aquaporin genes in Gossypium hirsutum L. and their expression profiling in the different tissues. Based on the bioinfor-matic analysis of the ESTs from the cotton fiber SSH-cDNA library, we obtained a cDNA sequence with an ORF of 927 bp, which has the homolog sequence of aquaporin genes, and it was designated as GhAQP2. With the protein sequence of GhAQP2 as a probe, two homologous ESTs in G. hirsutum database were screened out, and the rapid-amplification of cDNA ends (RACE) was used to amplify the full length cDNAs of GhAQP genes (designated as GhAQP3 and GhAQP4). Bioinformatic analysis showed that GhAQP2 and GhAQP3 were composed of four exons and three introns, GhAQP4 had three exons and two introns in com-parison of their sequences of genomic DNA and cDNA by DNAMAN. GhAQP2, GhAQP3, and GhAQP4 exhibited a typical structure with six transmembrane domains which linked by five loops and had the consensus sequence HINPAVTFG of MIP fam-ily and two highly conserved peptides Asn-Pro-Ala (NPA). These deduced amino acids showed high identity with the aquaporin of PIP2 subfamily reported from other plant species by multiple alignment analysis. The tissue specific expression analysis by using quantitative RT-PCR assays indicated that GhAQP2 expressed predominantly in 20 DPA fiber cell, while GhAQP3 expressed highly in the hypocotyl and cotyledon. Moreover, GhAQP4 showed highly significant expression level in 10 DPA fiber cell, hy-pocotyl and cotyledon. These results suggested that the three GhAQP genes might play different roles in different cotton tissues. The result of GhAQP2 predominantly expressed in 20 DPA fiber cell, provides important information for the further research on the gene-expression regulation during developmental stages from fiber cell elongation to secondary cell wall synthesis.关键词
陆地棉(Gossypium hirsutum L.)/棉纤维/水孔蛋白基因/序列分析/结构分析/表达模式Key words
Gossypium hirsutum L./Cotton fiber/Aquaporin genes/Sequence analysis/Structure analysis/Expression patterns引用本文复制引用
李伟,王涛,白志川,袁有禄,商海红,王少干,范森淼,李俊文,刘爱英,石玉真,龚举武,巩万奎..三个陆地棉水孔蛋白基因的克隆与表达分析[J].作物学报,2013,(2):222-229,8.基金项目
本研究由国家自然科学基金项目(31000732)和中央级公益性科研院所基本科研业务专项(SJB1009)资助 （31000732）
