摘要
低温诱导的富甘氨酸RNA结合蛋白GR-RBP3表达与采后黄瓜(Cucumis sativus)耐冷性密切相关。低温处理明显上调采后黄瓜的CsGR-RBP3表达,表明该基因是一个低温诱导型基因。为探究低温处理对CsGR-RBP3表达和启动子活性的诱导作用,本研究以黄瓜基因组DNA为模板,克隆了CsGR-RBP3开放阅读框上游1541 bp的启动子序列。生物信息学分析发现,克隆的启动子序列中含有真核生物典型的核心启动子元件TATA-box和CAAT-box,表明成功克隆了CsGR-RBP3启动子;含有低温响应元件LTR,推测低温可能通过影响CsGR-RBP3启动子活性激活CsGR-RBP3表达。启动子活性分析表明,CsGR-RBP3启动子能增强LUC活性,且能驱动GUS基因表达,表明该启动子具有启动基因转录的活性。在烟草和拟南芥中的转基因试验结果显示,低温处理明显提高了GUS活性和编码基因的表达,表明低温处理能增强CsGR-RBP3启动子活性。综之,低温通过诱导启动子活性进一步增强CsGR-RBP3的转录。
The expression of glycine-rich RNA-binding protein GR-RBP3 induced by low temperature was closely associated with the chilling tolerance of harvested cucumbers.Low temperature obviously upregulated CsGR-RBP3 expression in harvested cucumbers,suggesting it was a low temperature induced gene.To explore the inducing effect of low temperature on CsGR-RBP3 expression and its promoter activity,the promoter sequence at upstream of open reading frame of CsGR-RBP3 with the length of 1541 bp was cloned using DNA as template.The results of bioinformatics analysis showed that the promoter contained typical eukaryotic core elements TATA-box and CAAT-box,suggesting that CsGR-RBP3 promoter was successfully cloned.The low temperature response element LTR was also found,implying that low temperature might induce the expression of CsGR-RBP3 by affecting its promoter activity.Promoter activity analysis showed that CsGR-RBP3 promoter could enhance LUC activity and drive GUS gene expression,suggesting that the promoter had ability to initiate gene transcription.Transgenic experiments in tobacco and Arabidopsis showed that low temperature obviously enhanced GUS activity and its coding gene’s expression,indicating that low temperature enhanced the CsGR-RBP3 promoter activity.In conclusion,low temperature could induce the CsGR-RBP3 expression by enhancing its promoter activity.
作者
王斌
黄泳谚
易景怡
原远
Wang Bin;Huang Yongyan;Yi Jingyi;Yuan Yuan(Guangdong Provincial Key Laboratory of Utilization and Conservation of Food and Medicinal Resources in Northern Region,Shaoguan 512005,China;Henry Fok School of Biology and Agriculture,Shaoguan University,Shaoguan 512005,China)
出处
《山东农业科学》
北大核心
2022年第7期15-23,共9页
Shandong Agricultural Sciences
基金
广东省基础与应用基础研究基金资助项目(2020A1515010068,2022A1515011913)
广东省教育厅青年创新人才项目(2018KQNCX231)。
