摘要
二氨基庚二酸脱羧酶(diaminnopimelate decarboxylase, DAPDC)是合成赖氨酸的关键酶,在生物体的生命活动及非生物胁迫应答过程中发挥重要作用。本研究在砂藓(Racomitrium canescens)高温胁迫蛋白质组数据中发现一个表达量上调的DAPDC蛋白质,将其命名为RcDAPDC,并对编码此的基因全长CDS序列进行了克隆,还对此基因进行了基本的生物信息学分析,同时对砂藓在60℃高温胁迫、脱水和复水条件下的表达变化进行了分析。RcDAPDC基因CDS全长1 521 bp,编码蛋白质的氨基酸数为506,相对分子质量为55.9 kD,理论等电点为5.83,不稳定指数为36.91,是一个稳定蛋白,亲水性的平均值(GRAVY)为-0.169,预测该蛋白质为亲水性蛋白,不具有跨膜结构域。实时荧光定量PCR结果显示,RcDAPDC基因在60℃高温处理1 h时表达量最高;RcDAPDC基因在干旱和复水过程中呈现差异性表达,推测RcDAPDC不仅可能参与砂藓的抗高温胁迫反应,而且还可能参与砂藓的干旱胁迫响应和复水过程。本研究对进一步研究砂藓RcDAPDC基因的生物学功能提供了一定的理论基础和研究方向。
Diaminnopimelate decarboxylase(DAPDC) is a key enzyme in the synthesis of lysine and plays an important role in the life activities and abiotic stress responses of organisms. In this study, a DAPDC protein with up-regulated expression was found in the high temperature stress proteomics data of Racomitrium canescens, and the protein was named RcDAPDC. The coding sequence of the gene coding RcDAPDC was cloned, and bioinformatics analysis, expression analysis under 60℃ high temperature stress, desiccation and rehydration processes in R. canescens was carried on. The coding sequence of RcDAPDC gene is with a full length of 1 521 bp, encodes a protein with amino acids residues of 506, with a relative molecular mass of 55.9 kD, a theoretical isoelectric point of 5.83, and an instability index of 36.91, showing it is a stable protein. RcDAPDC was also predicted with a mean hydrophilicity(GRAVY) of-0.169, the protein is predicted as a hydrophilic protein and with no transmembrane domains. The real-time quantitative PCR results showed that RcDAPDC performed the highest expression level when treated with 60℃ for 1 h, and was also differentially expressed during desiccation and rehydration processes in R. canescens. It is speculated that RcDAPDC not only participates in the high temperature stress responses of R.canescens, but also may participate in the drought stress responses and rehydration processes. This study provides theoretical foundation and directions for further researches on the biological functions of the RcDAPDC in R.canescens.
作者
沙伟
王鑫
张梅娟
彭疑芳
马天意
Sha Wei;Wang Xin;Zhang Meijuan;Peng Yifang;Ma Tianyi(The Key Laboratory of Resistance Genetic Engineering and Coldland Biodiversity Conservation,College of Life Sciences,Agriculture and Forestry,Qiqihar University,Qiqihar,161006;College of Life Sciences,Northeast Agricultural University,Harbin,150030)
出处
《分子植物育种》
CAS
CSCD
北大核心
2020年第24期8092-8098,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)
黑龙江省人力资源和社会保障厅2018年省级留学回国人员择优资助项目共同资助。
