作物学报2013，Vol.39Issue(7)：1164-1171,8.DOI:10.3724/SP.J.1006.2013.01164
一个棉花GDSL脂肪酶基因的克隆与功能分析
Molecular Cloning and Functional Analysis of a GDSL Lipase Gene from Gossypium hirsutum L.
摘要
Abstract
GDSL and GXSXG lipases are two important sub-families of lipases.Unlike the GXSXG motif-containing lipases,GDSL lipases have a GDSL motif and are active in hydrolysis and synthesis of abundant ester compounds.In this experiment,a EST sequence (GenBank accession number:DR458916) was screened based on extremely significant expression differences at fiber initiation stages between lintless-fuzzless XinWX and linted-fuzzless XinFLM isogenic lines by 29K cotton genome array hybridization.Using the EST sequence as queries,the Gossypiurn EST database (http://www.ncbi.nlm.nih.gov/) was screened and the corresponding cDNA sequences containing a complete ORF were assembled.Further,the ORF information was reconfirmed in transcriptional and genomic level.As a result,a novel gene encoding GDSL lipase was cloned,and the gene was designated as GhGDSL (Gossypium hirsutum GDSL; GenBank accession number:KC186125).GhGDSL included an open reading frame of 1065 bp that encoded a polypeptide of 354 amino acids.The genome sequence indicated that GhGDSL had four introns and five exons.The homolog of GhGDSL 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.Sequence alignment from different cotton species showed GhGDSL homoelogs in tetraploid were evolved independently,with more variations in D-subgenome than in A-subgenome.GhGDSL homoelogs in tetraploid were located on chromosome 4 (Chr.A4) and chromosome 22 (Chr.D4) by developing subgenome-specific SNP marker,respectively.Q-PCR analysis showed that GhGDSL was accumulated highly in ovule and fiber tissue at 3-10 days post anthesis (DPA),with expression peak at 8DPA in Hai 7124 and high expression from 5DPA to 10 DPA in TM-1.The association analysis between GhGDSL and the traits related to fiber and seed qualities in corresponding [(TM-1 ×Hai 7124)×TM-1] BC1S1 individuals showed that GhGDSL homolog in A-subgenome was significantly correlated with the seeds' fat content (P=0.048),and GhGDSL homolog in D-subgenome was very significantly correlated with the seeds' protein content (P=-0.008).These results suggested that GhGDSL might be functionally important in the metabolic process of lipid and protein in seeds and in the process of fiber development.关键词
GDSL脂肪酶/克隆/表达/基因定位/关联分析/棉花Key words
GDSL lipase/ Cloning/ Expression/ Gene mapping/ Association analysis/ Cotton引用本文复制引用
佟祥超,王丽曼,胡文静,张雪颖,张天真,郭旺珍..一个棉花GDSL脂肪酶基因的克隆与功能分析[J].作物学报,2013,39(7):1164-1171,8.基金项目
本研究由国家重点基础研究发展计划(973计划)项目(2011CB109300)资助. （973计划）
