小麦TaPSKR1基因的克隆与表达分析

Cloning and Expression Analysis of TaPSKR1 Gene in Wheat

植物磺化肽激素受体(PSK receptor, PSKR)在促进植物细胞增殖和非生物胁迫中起着重要作用。为了探讨小麦PSKR的序列特征和生物学功能,采用同源克隆技术,从普通小麦品种晋麦47根组织中克隆出TaPSKR1 3个部分同源基因的cDNA序列,因3个基因分别位于6A、6B和6D染色体上,故分别命名为TaPSKR1-6A、TaPSKR1-6B和TaPSKR1-6D。并且利用生物信息学手段对其基因结构、蛋白理化性质、顺式作用元件、功能结构域及系统进化树进行分析,通过qRT-PCR分析TaPSKR1基因在不同组织以及不同逆境胁迫下的表达模式。结果表明,TaPSKR1-6A、TaPSKR1-6B和TaPSKR1-6D均包含一个外显子,其开放阅读框分别为3 153,3 132,3 156 bp,分别编码1 050,1 043,1 051个氨基酸。生物信息学分析结果表明,TaPSKR1定位在细胞膜上,具有信号肽、跨膜结构域和8个LRRs结构域及胞内激酶结构域,属于PSKR家族成员。系统进化显示,TaPSKR1蛋白与小麦近缘物种及水稻亲缘关系较近,处于同一分支上。实时定量PCR分析结果表明,TaPSKR1基因在根、茎、叶片、穗和种子中均有表达,在根中的表达量极高;逆境胁迫分析表明,干旱和盐胁迫处理下,叶片中TaPSKR1的3个部分同源基因的表达急剧上调,推测TaPSKR1可能在小麦抵抗逆境胁迫过程中起重要的调控作用。

Phytosulfokine receptor(PSKR)plays an important role in promoting plant cell proliferation and is involved in plant response to abiotic stresses.To explore the sequence characteristics and the function of wheat PSKR genes,the cDNA sequences of three homologous genes of TaPSKR1 were cloned from wheat variety Jinmai 47 by homologous cloning technology,named TaPSKR1-6A,TaPSKR1-6B and TaPSKR1-6D because of their locations on chromosome 6A,6B and 6D,respectively.The gene structure,protein physical and chemical properties,cis acting elements,functional domains and evolutionary relationships were analyzed by bioinformatics analysis.The expression patterns of TaPSKR1 genes in different tissues and under different stresses were detected by qRT-PCR.The results showed that TaPSKR1-6A,TaPSKR1-6B and TaPSKR1-6D all contained one exon.The open reading frame(ORF)of the three TaPSKR1 genes were 3153,3132,3156 bp,respectively,which encoded 1050,1043 and 1051 amino acid residues.Bioinformatics analysis showed that TaPSKR1 proteins were located on the cell membrane,containing signal peptide,transmembrane domains,eight LRRs type domains and intracellular kinase domain,which belonged to PSKR gene family.Phylogenetic analysis showed that TaPSKR1 proteins had closely relationship with its related species and rice,which were clustered into the same subgroup.The results of expression analysis showed that TaPSKR1 genes were expressed in roots,stems,leaves and seeds,and the expression levels in roots were the highest.Under drought and salt stress treatments,the expressions of three homologous copies of TaPSKR1 genes were sharply upregulated in leaves,suggesting that TaPSKR1 might play an important regulatory role in wheat defense to abiotic stresses.