陆地棉GhSDP1及其启动子的克隆与表达分析

三酰基甘油脂肪酶(SDP1)是催化三酰甘油降解的关键酶,在植物油脂代谢调控中起着重要作用。克隆棉花SDP1并研究其在3种胁迫下的表达分析,为解析棉花SDP1的生物学功能提供依据。以陆地棉品种冀丰1271为试材,克隆GhSDP1编码序列和上游启动子序列;利用PlantCARE分析GhSDP1启动子区顺式作用元件;qRT-PCR检测逆境胁迫下GhSDP1的表达谱;通过烟草瞬时表达pGhSDP1启动子+GUS载体检测启动子活性。结果表明,GhSDP1的编码序列为2 541 bp,其在盐、低温和干旱胁迫下呈差异表达模式。pGhSDP1除具有启动子所必需的TATA-box和CAAT-box等基本顺式作用元件外,还含有多个与光响应、激素响应及逆境应答等相关的顺式作用元件。棉花pGhSDP1启动子能驱动GUS蛋白高效表达,具有较强的启动子活性。研究揭示了棉花GhSDP1参与胁迫应答的新功能。

Cloning and Expression Analysis of GhSDP1 and Its Promoter in Gossypium hirsutum

Triacylglycerol lipase(SDP1)is a key enzyme in the degradation of triacylglycerol and plays an important role in the regulation of vegetable oils.We focused on the cloning of the cotton SDP1 gene and tits expression analysis under three stresses,which provides basis for analyzing the biological function of the cotton SDP1 gene.The coding sequence and upstream promoter sequence of GhSDP1 were cloned in cotton variety Jifeng 1271.PlantCARE was used to analyze the cis-acting elements in the promoter region of GhSDP1,and qRT-PCR was applied to detect the expression profile of GhSDP1 under stress.The promoter activity analysis of pGhSDP1+GUS fusion vector was carried out by tobacco transient expression.The results showed that the coding sequence of GhSDP1 was 2 541 bp,and the gene presented a differential expression pattern under salt,low temperature and drought stress.pGhSDP1 not only had the basic cis-acting elements such as TATA-box and CAAT-box,but also contained a number of cis-acting elements related to light response,hormone response and adversity response.The pGhSDP1 promoter in cotton can drive the high-efficiency expression of GUS protein and has strong promoter activity.The study revealed that the cotton GhSDP1 gene participates in the corresponding new functions of the stress.