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[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。
[Objective] The research aimed to explore the molecular mechanism of ethylene insensitive flower senescence. [Method] The petals of dahlia petals were used as materials to separate and identify the senescence-related proteins and their coding genes by using two-dimensional electrophoresis, mass spectrometry and molecular biology techniques. [Result] From the two-dimensional electrophoresis patterns of the petal proteins in the stages of transpiration, full bloom and decay of dahlia, 44 protein spots with more than 2 times of expression differences were detected, and the glucosyltransferases The expression level of Xyloglucan endotransglycosylase / hydrolase (XTH), which increases with the senescence process of petals, belongs to an aging-related protein. Using Dahlia petal RNA as a material, a pair of degenerate primers was designed and the cDNA sequence of XTH gene from Dahlia flower petal was cloned by RTPCR. The full-length coding sequence of Dahlia petal was 882 bp in length and encoded 293 amino acid residues (Gene Accession No. HM053613.1 , Named it Dp XTH1). Cluster analysis showed that the amino acid sequence of Dp XTH1 has high homology with XTH of other plants. [Conclusion] The isolated and identified Dahlian Dp XTH1 belonged to the XTH family of plants, and its biological function was related to the senescence process and regulation of the petals of dahlia.