论文部分内容阅读
脱水素(DHN)在植物经受逆境胁迫条件下对于保护细胞内蛋白质和膜结构具有重要作用。本研究以芍药‘大富贵’芽为试材,采用RT-PCR技术克隆得到1个脱水素基因PlDHN2(Gen Bank登录号KY272747),该基因具有399 bp的开放阅读框,编码132个氨基酸,具有DHN家族特有的Y、S和K片段,属于典型的Y_3SK_2型脱水素;基因结构分析表明,PlDHN2具有1个长度为84 bp的内含子,位于S片段。以拟南芥和大麦的脱水素蛋白分类为参照,聚类分析结果表明,PlDHN2与Y_nSK_2型脱水素聚合在一起,进化关系较近。半定量PCR分析显示,PlDHN2在芍药各器官中均有表达,其中在芽中的表达最高。亚细胞定位观察结果显示:PlDHN2蛋白定位于细胞质膜和细胞核中。表达特性分析表明,PlDHN2在低温、高温、水淹和ABA胁迫条件下,表达量均不同程度地上调,说明该基因可能参与了芍药对低温等非生物胁迫的耐受调节过程。
Dehydratase (DHN) plays an important role in protecting intracellular proteins and membrane structures under stress conditions in plants. In this study, peony ’big rich’ bud as the test material, cloned by RT-PCR to obtain a dehydrin gene PlDHN2 (Gen Bank accession number KY272747), the gene has a 399 bp open reading frame, encoding 132 amino acids, with The Y, S and K fragments belonging to the DHN family belong to the typical Y_3SK_2-type dehydratin. Gene structure analysis showed that PlDHN2 has an 84-bp intron located in the S-segment. Based on the classification of the dehydrins of Arabidopsis and barley, the clustering analysis showed that the aggregates of PlDHN2 and Y_nSK2 were aggregated and their evolutionary relationships were close. Semi-quantitative PCR analysis showed that PlDHN2 was expressed in all organs of Paeonia lactiflora, among which the highest expression was found in shoots. The results of subcellular localization showed that the PlDHN2 protein localized in the plasma membrane and nucleus. Expression analysis showed that the expression level of PlDHN2 was up-regulated to some extent under the conditions of low temperature, high temperature, flooding and ABA stress, indicating that this gene may be involved in the tolerance regulation of Paeonia lactiflora to abiotic stresses such as low temperature.