棉花GhTFL1b基因的表达及调控分析

Expression and Regulation Analysis of GhTFL1b Gene in Upland Cotton

为解析棉花中磷脂酰乙醇胺结合蛋白GhTFL1b基因对胁迫的响应及生理功能,从棉花中克隆了磷脂酰乙醇胺结合蛋白GhTFL1b基因,并对该基因进行表达分析和蛋白质互作研究。利用启动子分析软件PlantCARE预测得出,GhTFL1b启动子区域有茉莉酸响应元件、脱落酸响应元件、干旱诱导的MYB结合位点和光周期响应元件等。对GhTFL1b进行组织特异性表达分析,发现该基因在花中表达量最高,其次是根和顶芽,其他组织中表达量极低。进一步比较栽培种和半野生种不同时期的表达,发现无明显差异。激素和胁迫处理研究表明,GhTFL1b基因受水杨酸(Salicylic acid,SA)诱导低调表达,而受盐胁迫处理后高调表达。酵母双杂交验证GhFD与GhTFL1亚组蛋白互作,结果表明GhFD蛋白只能与GhTFL1b互作,而不与GhTFL1a、GhTFL1b、GhTFL1c互作。GhTFL1b基因不参与光周期调控通路,可能参与植物逆境胁迫水杨酸和盐胁迫响应的调控。

In order to study the response of cotton phosphatidylethanolamine-binding protein GhTFL1b gene to stress and physiological function,we cloned the GhTFL1b gene from upland cotton,and then analyzed its expression pattern and protein interaction. The results of promoter structure prediction using PlantCARE software revealed that GhTFL1b promoter contained MeJA responsiveness element,abscisic acid(ABA)responsiveness element ,MYB binding sites involved in drought inducibility and light responsiveness elements. Meanwhile,Quantitative Real time PCR result indicated that the expression profile of GhTFL1b was induced by salicylic acid(SA)negatively,while it could be regulated by salt(NaCl)positively. Yeast two hybrid approach were under taken to elucidate the protein interaction of GhFD and GhTFL1 subgroup,and the experiments showed that GhFD could interact with GhTFL1b but not with GhTFL1a,GhTFL1b and GhTFL1c in cotton. Taken together,the result suggested that GhTFL1b might be involved in the regulation of response to abiotic stresses(SA and salt).