沙柳SpsLAZY1a、SpsLAZY1b基因启动子克隆及表达分析

Cloning of SpsLAZY1a and SpsLAZY1b Gene Promoters from Salix psammophila and Their Expression Analysis

为研究SpsLAZY1a、SpsLAZY1b基因的表达调控机制,本研究从沙柳中分离了SpsLAZY1a、SpsLAZY1b基因的启动子片段。使用PlantCARE数据库对该启动子片段中顺式作用元件预测显示,SpsLAZY1a和SpsLAZY1b启动子序列中除了包含核心元件CAAT-box和TATA-box外,还含有光、茉莉酸甲酯、脱落酸、低温、乙烯、赤霉素等响应元件。Pro_(SpsLAZY1a)和Pro_(SpsLAZY1b)在烟草中瞬时表达和转基因84K杨中的GUS显色结果表明,Pro_(SpsLAZY1a)和Pro_(SpsLAZY1b)具有启动子活性。对转基因84K杨的GUS酶活检测结果表明,Pro_(SpsLAZY1a)的GUS表达活性强于Pro_(SpsLAZY1b)。对转基因84K杨茎的切片结果显示,蓝色显色反应主要集中于内皮层和韧皮部。上述结果表明,SpsLAZY1a、SpsLAZY1b基因的启动子是非组织特异性表达启动子,并且这两个基因启动子的活性存在差异。本研究结果为解析LAZY基因的上游调控机制提供参考。

To study the transcriptional regulation mechanism of SpsLAZY1a and SpsLAZY1b genes,their promoter fragments were cloned from Salix psammophila C.Wang&C.Y.Yang.By analyzing cisacting element in the promoter sequences using plantCARE database,the promoter sequences of both genes contain the core elements CAAT-box and TATA-box,and the elements responding to light,methyl jasmonate,abscisic acid,ethylene,gibberellin,and low temperature.GUS staining in tobacco transient expression and stable transgenic 84K poplar revealed the transcriptional activity of both promoters Pro_(SpsLAZY1a) and Pro_(SpsLAZY1b).The Pro_(SpsLAZY1a) activity was stronger than that of Pro_(SpsLAZY1b )in transgenic 84K poplar.The results of stem basal sections of transgenic 84K poplar showed that the expression of Pro_(SpsLAZY1a) and Pro_(SpsLAZY1b) was mainly observed in endothelium and phloem.The promoters of SpsLAZY1a and SpsLAZY1b genes were non-tissue-specific,and their activities of the promoters were different.Collectively,this study provides a reference for analyzing the regulation mechanism of LAZY gene.