‘凤丹’牡丹PoERF113基因的克隆及表达分析  

Cloning and expression analysis of PoERF113 gene in Paeonia ostii‘Fengdan’

作　　者：魏祯祯 宋程威[1] 郭丽丽[1] 郭琪[1] 侯小改[1] WEI Zhenzhen;SONG Chengwei;GUO Lili;GUO Qi;HOU Xiaogai(College of Agriculture,Henan University of Science and Technology,Luoyang,Henan 471023,China)

机构地区：[1]河南科技大学农学院,河南洛阳471023

出　　处：《湖南农业大学学报（自然科学版）》2022年第6期659-665,共7页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(U1804233);河南省创新型科技人才队伍建设工程(202101510003);河南省中药材产业技术体系(2021-24)。

摘　　要：为克隆‘凤丹’(Paeonia ostii)花瓣衰老相关基因PoERF113的序列,分析其编码蛋白性质,探索其在不同发育时期花瓣、不同组织和不同品种的表达模式及对乙烯的响应,采用RT-PCR技术,从‘凤丹’牡丹中克隆到花瓣衰老相关基因PoERF113。该基因属于AP2/ERF家族,其开放阅读框(ORF)为636bp,编码1个由211个氨基酸残基组成的蛋白,含有1个AP2保守结构域,不包含跨膜结构,蛋白相对分子质量约为53000,理论等电点(pI)为5.13,为不稳定蛋白,具有亲水性;qRT-PCR分析结果表明,PoERF113基因在露色期、初开期、半开期、盛开期、始衰期、衰败期均有表达,主要在‘凤丹’露色期的花瓣和叶片中表达;在早花‘凤丹’突变株系中表达量最高;PoERF113基因在早花‘凤丹’突变株系与‘凤丹’中的序列相同,不存在碱基差异;PoERF113基因对乙烯有响应,其表达量随处理时间和生育进程的延长呈逐渐升高的趋势,推测该基因可能与乙烯诱导后导致花衰老进程相关。To investigate the petal senescence-related gene PoERF113 of‘Fengdan’(Paeonia ostii)expression patterns in petals,different tissues and different varieties at different flowering stages and their response to ethylene,we did cloning analyzed its coded protein properties in this study.First,we used RT-PCR technology to clone the petal senescence related gene PoERF113 gene from‘Fengdan’.The gene belongs to AP2/ERF family.Its open reading frame(ORF)is 636bp,encoding a protein consisting of 211 amino acid residues.It contains one AP2 conservative domain and does not contain transmembrane structure.The molecular weight of the protein is about 53000,and the theoretical isoelectric point(pI)is 5.13.It is unstable protein and hydrophilic.The results of qRT-PCR analysis showed that PoERF113 gene was expressed in the open color stage,the first open stage,the half open stage,the full bloom stage,the first decline stage,and the decline stage,mainly in the petals and leaves of‘Fengdan’in the open color stage;The highest expression was found in‘Fengdan’mutant strain of Zaohua;The sequence of PoERF113 gene in the‘Fengdan’mutant line of Zaohua was the same as that in‘Fengdan’and there was no base difference;PoERF113 gene was responsive to ethylene,and its expression level gradually increased with the extension of treatment time and growth process.It was speculated that this gene might be related to the process of flower senescence induced by ethylene.This experiment laid a preliminary foundation for further study on the function of PoERF113 gene and its molecular mechanism in peony petal senescence and florescence regulation.

关 键 词：‘凤丹’牡丹 PoERF113 基因克隆 表达分析 

分 类 号：Q786[生物学—分子生物学] S685.11[农业科学—观赏园艺]