砂藓抗旱相关基因磷脂酶D的克隆和表达分析  

Cloning and Expression Analysis of Drought-resistant Related Phospholipase D Gene from Racomitrium canescens

作　　者：沙伟[1] 张时通 朱巍巍[1] 张梅娟[1] 马天意 Sha Wei;Zhang Shitong;Zhu Weiwei;Zhang Meijuan;Ma Tianyi(The Key Laboratory of Resistance Genetic Engineering and Coldland Biodiversity Conservation,College of Life Sciences,Agriculture and Forestry,Qiqihar University,Qiqihar,161006)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,抗性基因工程与寒地生物多样性保护黑龙江重点实验室,齐齐哈尔161006

出　　处：《分子植物育种》2021年第1期123-129,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31570207);黑龙江省省属高等学校基本科研业务费青年创新人才项目(135309364);黑龙江省省属高等学校基本科研业务费科研项目(YSTSXK201876);黑龙江省人力资源、社会保障厅2018年省级留学回国人员择优资助项目共同资助。

摘　　要：磷脂酶D是广泛存在于植物中的一种磷脂酶,在细胞结构组成和生理功能的变化中具有非常重要的作用。在本研究中,以在砂藓(Racomitrium canescens)转录组数据库中获得的一条与抗旱相关的磷脂酶D基因为基础,采用RT-PCR技术克隆得到该基因的全长CDS序列,命名为Rc PLD。生物信息学分析结果表明,Rc PLD基因c DNA全长1 420 bp,含有894 bp的开放阅读框,能够编码长度为297个氨基酸的多肽链。Rc PLD蛋白质的生物信息学分析结果表明,Rc PLD相对分子质量为33.192 k D,理论等电点p I为6.60,不含有跨膜结构,存在信号肽,具有磷脂酶D的保守结构域,属于不稳定的疏水性蛋白质。实时荧光定量PCR分析表明,在砂藓的复水过程和脱水处理条件下Rc PLD基因均有差异表达,说明该基因可能参与砂藓中响应干旱胁迫的某些生理代谢途径。本研究可为苔藓植物中抗旱机制的研究提供理论基础。Phospholipase D(PLD)is a phospholipase widely presents in plants,which plays a vital role in the changes in cellular structural composition and physiological functions.In this study,coding sequence of a PLD gene was cloned from Racomitrium canescens transcriptomic database and named as Rc PLD.Bioinformatics analysis showed that the c DNA of Rc PLD gene was 1420 bp in length and 894 bp in open reading frame,encoding a polypeptide chain consisting of 297 amino acid residues.The bioinformatics analysis of Rc PLD protein showed that relative molecular mass of Rc PLD was 33.192 k D,the theoretical isoelectric point was 6.60,with no transmembrane structure,containing the signal peptide,and a conserved domain of phospholipase D,which was also an unstable hydrophobic protein.Results of real-time q PCR analysis showed that Rc PLD was differentially expressed under rehydration and dehydration treatment conditions of R.canescens,indicating that Rc PLD may be involved in some physiological metabolic pathways in R.canescens in response to drought stress.This study may provide a theoretical basis for the researches of drought resistance mechanisms in bryophytes.

关 键 词：砂藓(Racomitrium canescens) 磷脂酶D 脱水处理 复水处理 表达分析 

分 类 号：Q943.2[生物学—植物学]