摘要
从红肉火龙果果肉中克隆得到3个ERF转录因子,命名为HpERF1/2/3。氨基酸序列比对和进化树分析发现HpERF1/2/3具有典型的AP2/ERF结构域,ORF长度分别为1 020、843、1 269 bp,编码长度分别为339aa、280aa、422aa,分子量为37.4、31.99、45.08 k Da,以及等电点为5.69、6.36、6.93的氨基酸多肽链。实时荧光定量聚合酶链式反应表明,随着红肉火龙果果实发育,HpERF1/2/3基因的表达明显增强。亚细胞定位和转录活性分析显示HpERF1/2/3都定位于细胞核,是核蛋白,并且具有转录激活活性。双荧光素酶瞬时表达分析显示,HpERF1/2/3可以激活红肉火龙果甜菜红素合成途径上关键基因HpCyt P450-like1的启动子活性。这些结果表明HpERF1/2/3可能通过调控甜菜红素合成相关基因表达来参与红肉火龙果果实品质形成过程,加深了对红肉火龙果果实品质生物学的认识。
In the present work, three ERF transcription factors（TFs） HpERF1/2/3 were isolated from pitaya fruit（Hylocereus polyrhizus）. The deduced amino acid sequence alignment and phylogenetic tree analysis indicated that,HpERF1/2/3 contained a typical AP2/ERF domain, the lengths of open reading frames（ORF） were 1 020 bp, 843 bp and 1 269 bp encoding 339, 280 and 422 amino acids respectively, and their molecular weight were 37.4 k Da,31.99 k Da and 45.08 k Da with p I value of 5.69, 6.36 and 6.93 respectively. Real-time quantitative PCR displayed that, HpERF1/2/3 were up-regulated following pitaya fruit（Hylocereus polyrhizus） development. Sub-cellular localization and transcriptional activity analysis revealed that, HpERF1/2/3 were nuclear proteins with transcriptional activation activity. In addition, dual-luciferase transient assay demonstrated that, HpERF1/2/3 could activate the promoter activities of key gene HpCyt P450-like1 in the biosynthetic pathway of betalains of red pitaya fruit. Taken together, these results suggested that, HpERF1/2/3 might be involved in pitaya fruit development via regulating the expression of genes related to betalain biosynthesis, which deepens the understanding of transcriptional regulation mechanism of pitaya fruit development.
出处
《保鲜与加工》
CAS
北大核心
2018年第1期32-40,共9页
Storage and Process
