摘要
【目的】甘油-3磷酸酰基转移酶(GPAT)是植物三酰甘油合成途径的3个关键酶之一,在油脂品质改良和生物质能源利用方面具有重要的应用价值。【方法】本研究通过同源克隆技术,从多年连续自交的海岛棉材料海资8号中获得了海岛棉甘油-3磷酸酰基转移酶基因,命名为GbGPAT2。【结果】GbGPAT2基因cDNA全长1294 bp,包含1个1113 bp的开放阅读框,编码1条370个氨基酸组成的多肽,预测分子量为42.4 kD,等电点为8.72。该基因编码的蛋白具有2个保守结构域,即典型的溶血磷脂酰基转移酶相似家族保守区(LPLAT-LPCAT1-like,Lysophospholipid Acyltransferases-Lysophospholipid Acyltransferases like)和酰基转移酶超家族保守结构域(NAT-SF,N-Acyltransferase superfamily)。GbGPAT2与小桐子、杨树、大豆和拟南芥的GPATs氨基酸同源性在91.2%~85.4%。跨膜区分析表明,GbGPAT2有2个跨膜结构域,亚细胞定位试验表明,GbGPAT2定位于细胞质膜上。qRT-PCR分析表明,GbGPAT2在胚发育的不同时期表达丰度不同,在开花后25 d的胚中表达量最大。【结论】推测GbGPAT2在海岛棉种子油脂合成中起重要作用。
【Objective】GPAT is the first enzyme in the plant glycerolipid synthetic pathway,which may play an important role in oil quality improvement and biomass energy.【Methord】Based on the GPATs obtained from GenBank,ESTs of Gossypium were selected to obtain the cDNA sequence of GbGPAT2 gene by RT-PCR from Haizi 8,which was a homozygous line of Gossypium barbadense.【Result】The full length cDNA of GbGPAT2 was 1294 bp,containing an open reading frame(ORF)of 1113 bp,which encoded a protein of 370 amino acids.Its molecular weight was about 42.4 kDa,and pI 8.72.GbGPAT2 contained a typical conserved LPLAT-LPCAT1-like domain and a NAT-SF superfamily domain.GbGPAT2 putative protein shared 87%-92%identity with Jatropha curcas and Arabidopsis,respectively.GbGPAT2 shared only 8%identity with GhGPAT(ADK23938.1,cloned from upland cotton without further information).The bioinformatics analysis of the membranes and subcellular localization showed that GbGPAT2 might be located in the cytoplasm membrane or the chloroplast mem-brane,with two transmembrane domain(TMD)structures,subcellular localization analysis showed that GbGPAT2-GFP fusion protein was distributed specifically in membranes.The embryo specific expression analysis by using quantitative RT-PCR assay indicated that the expres-sion level of GbGPAT2 reached a peak in the stage 25 days post anther(DPA).【Conclusion】As a result,GbGPAT2 may play an important role in the Gossypium barbadense seed oil synthesis.
作者
甄军波
刘琳琳
杜海英
刘迪
蔡肖
张建宏
迟吉娜
ZHEN Jun-bo;LIU Lin-lin;DU Hai-ying;LIU Di;CAI Xiao;ZHANG Jian-hong;CHI Ji-na(Cotton Research Institute,Hebei Academy of Agriculture and Forestry Sciences,Key Laboratory of Cotton Biology and Genetic Breeding in Huanghuaihai Semiarid Area,Ministry of Agriculture,Hebei Shijiazhuang 050051,China)
出处
《西南农业学报》
CSCD
北大核心
2020年第3期503-508,共6页
Southwest China Journal of Agricultural Sciences
基金
河北省现代农业科技创新工程子课题(2019-3-7-3)
河北省棉花产业技术体系(HBCT2018040207)。
关键词
海岛棉
甘油-3磷酸酰基转移酶
基因克隆
表达分析
Gossypium barbadense
Glycerol-3-phosphate acyltransferase(GPAT)
Gene cloning
Expression analysis
