Identification of Senescence-associated Protein DpXTH1 and its Gene Cloning in Dahlia Petals  

大丽花花瓣衰老相关蛋白DpXTH1及其编码基因的分离（英文）

作　　者：张萍萍[1] 阚雪芹[1] 李坚[1] 陈驰[1] 罗成洋 谈建中[1] 

机构地区：[1]苏州大学建筑学院/苏州市建筑与城市环境重点实验室,江苏苏州215123

出　　处：《Agricultural Science & Technology》2015年第7期1490-1493,共4页农业科学与技术（英文版）

基　　金：Supported by Applied Basic Research Project of Suzhou City(SYN201405);National College Students’Innovative Entrepreneurial Training Program(201410285049)~~

摘　　要：[Objective] This study aimed to explore the molecular mechanism of senescence in ethylene-insensitive flowers. [Method] The dahlia petals were used as matedal, and the senescence-associated proteins were isolated and identified using two-dimensional electrophoresis, mass spectrometry and an encoding gene was cloned using molecular biology techniques. [Result] In the two-dimensional elec- trophorogram of proteins from dahlia petals at building color, full flowering and flow- er senescence pedods, a total of 44 protein spots with differences in expression level more than two times were detected. From the 44 protein spots, xyloglucan （XTHs）, a senescence-associated protein, was iso- lated and identified and its expression level was increased continuously with the senescence process of dahlia petals. By using the total RNA of dahlia petals as matedal and a pair of degenerate pdmers, the cDNA sequence of XTH gene was cloned by RT-PCR. The encoding region of XTH gene has a full length of 882 bp, encoding 293 amino acid residues, and is named as DpXTH1 （Accession number： HM053613.1）. The cluster analysis showed that the amino acid sequence of DpXTH1 has high homology with those of XTHs in other plants. [Conclusion] The isolated and identified DpXTH1 from dahlia petals belonged to the XTH family in plants, and its biological function was associated with the senescence process and regulation of dahlia petals.[目的]为了探讨乙烯非敏感型花卉衰老相关的分子机理。[方法]以大丽花花瓣为材料,利用双向电泳、质谱分析及分子生物学技术,分离、鉴定花瓣衰老相关蛋白质及其编码基因。[结果]从大丽花透色期、盛花期和衰败期花瓣蛋白质的双向电泳图谱中,检测到44个表达量差异在2倍以上的蛋白质斑点,从中分离、鉴定了木葡聚糖内糖基转移酶/水解酶(xyloglucan endotransglycosylase/hydrolase,XTH),其表达量随花瓣的衰老进程而持续增加,属于一种衰老相关蛋白质。以大丽花花瓣RNA为材料,设计一对简并引物,通过RTPCR技术克隆了大丽花花瓣XTH基因的c DNA序列,其编码区序列全长882 bp,编码293个氨基酸残基(基因登录号HM053613.1,命名其为Dp XTH1)。聚类分析表明,Dp XTH1氨基酸序列与其他植物的XTH具有较高的同源性。[结论]分离、鉴定的大丽花Dp XTH1属于植物XTH家族,其生物学功能与大丽花花瓣的衰老进程及调控有关。

关 键 词：DAHLIA PETALS Xyloglucan endotransglycosylase/hydrolase Senescence-associated protein Gene cloning 

分 类 号：S682.261[农业科学—观赏园艺]