摘要
[目的]研究油菜素内酯受体BRI1的同源基因(JcBRI1)在麻疯树花发育过程中的作用。[方法]利用RT-PCR技术克隆JcBRI1基因的CDS,以pET-30a(+)质粒为框架构建原核表达载体转化至大肠杆菌进行诱导表达,随后利用LC-MS/MS对表达产物进行质谱鉴定,并以氨基酸序列为基础分析JcBRI1蛋白质结构。利用荧光定量PCR分析麻疯树JcBRI1基因在雌雄花发育的关键时期的表达水平,初步确定其在麻疯树花发育过程中的表达模式。[结果]克隆获得了JcBRI1基因的CDS,长度为3 591 bp。SDS-PAGE检测结果表明:JcBRI1基因在大肠杆菌BL21(DE3)和Rosetta(DE3)中均能表达,但在低温环境的表达量更高。原核表达产物的LC-MS/MS鉴定结果表明:JcBRI1氨基酸序列与预期的一致,JcBRI1基因的表达产物为麻疯树BRI1蛋白。对JcBRI1在麻疯树花器官不同发育时期的表达水平进行检测分析,结果表明:JcBRI1基因的表达量在雌花发育的第一个时期即大孢子母细胞时期就达到了最高值,在随后几个时期持续下调;然而,其在雄花各个时期的表达量并无明显差异,表明其很有可能参与了麻疯树雌花大孢子母细胞发育过程。[结论]麻疯树JcBRI1基因为LRR-RLKs家族成员BRI1的同源基因,很有可能参与麻疯树雌花大孢子母细胞的发育过程,至于JcBRI1在麻疯树大孢子母细胞发育过程中的具体作用机制还需进一步研究。
[Objective] To study the function of the homologous gene of brassinolide receptor BRI1(JcBRI1) in the development of Jatropha curcas flower.[Method] The CDS of JcBRI1 gene was cloned by RT-PCR, and inserted into the prokaryotic expression vector pET-30 a(+). The prokaryotic expression protein was identified by LC-MS/MS, and the protein structure was analyzed by bioinformatics tools. The qRT-PCR was carried out to detect the transcriptional level of JcBRI1 in the critical developmental stages of flower.[Result] The length of JcBRI1 CDS was 3 591 bp, which was predicted to encode a piptide of 1 196 amino acids. The results of LC-MS/MS detection and protein structure analysis of prokaryotic expression products showed that this protein is J. curcas BRASSINOSTEROID INSENSITIVE 1. qRT-PCR was carried out to investigate the expressive patterns of JcBRI1 in different developmental stages of flower. It seems that higher transcriptional level of JcBRI1 is likely to promote the development of the megaspore mother cell in female flower.[Conclusion] The JcBRI1 gene is the homologous gene of brassinolide receptor protein BRI1, it probably participates in the division process of the megaspore mother cells during the flower development of J. curcas.
作者
雷世康
徐刚
LEI Shi-kang;XU Gang(Institute for Forest Resources and Environment of Guizhou, Guizhou University, Guiyang 550025, Guizhou, China;College of Life Science, Guizhou University, Guiyang 550025,Guizhou,China;Institute of Entomology , Guizhou University , Guiyang 550025 , Guizhou , China)
出处
《林业科学研究》
CSCD
北大核心
2019年第5期121-129,共9页
Forest Research
基金
国家自然科学基金“赤霉素对麻疯树雌雄花分化和发育调控的机制研究”(31760198)
