《植物生理学报》 2017, 53(7): 1297-1305

芍药脱水素基因PlDHN2的克隆及表达分析

陈煜^*, 马燕^*, 国静, 李俊杰, 韩璐璐, 郭先锋^**

山东农业大学林学院, 山东泰安271018

通信作者：陈；E-mail: guoxf@sdau.edu.cn

摘　要：

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 (GenBank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Y₃SK₂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_nSK₂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。

关键词：芍药; 脱水素; 克隆; 亚细胞定位; 表达分析

收稿：2017-05-12   修定：2017-06-23

资助：山东省自然科学基金(ZR2014CM028)。

Cloning and expression analysis of the dehydrin gene PlDHN2 in peony (Paeonia lactiflora)

CHEN Yu^*, MA Yan^*, GUO Jing, LI Jun-Jie, HAN Lu-Lu, GUO Xian-Feng^**

Colloge of Forestry, Shandong Agricultural University, Taian, Shandong 271018, China

Corresponding author: CHEN Yu; E-mail: guoxf@sdau.edu.cn

Abstract:

Dehydrin (DHN) proteins play a protective role in plants under different abiotic stress conditions. In this study, the full-length cDNA of a dehydrin-like gene PlDHN2 (GenBank accession number KY272747) was cloned from buds of Paeonia lactiflora ‘Dafugui’ by reverse transcription PCR (RT-PCR). PlDHN2 gene contains 399 bp open reading frame (ORF) encoded 132 amino acids. Sequence alignment and phylogenetic analysis showed that PlDHN2 posseses typical characteristic of DHN family, belonging to the typical Y3SK2-type dehydrin. Gene structural analysis demonstrated that PlDHN2 contains one 84 bp intron, which was inserted in the nucleotide sequence encoding the S-motif and characterised by a GT-AG border. Semi-quantitative PCR showed that PlDHN2 was expressed in each organ and extremely high in buds, but at very low levels in roots and petals. Subcellular localization analysis revealed that the PlDHN2 protein was mainly detected in the cytomembrane and nuclear. The expression of PlDHN2 was up-regulated in response to cold, heat, waterlogging and ABA treatment in various degree. It was deduced that PlDHN2 plays an important role in stress tolerance of cold, heat, waterlogging and ABA.

Key words: peony (Paeonia lactiflora); PlDHN2; molecular cloning; intron; subcellular localization; expression analysis