割手密SsWRKY1基因的克隆与表达分析

Cloning and Expression Analysis of the SsWRKY1 Gene in Saccharum spontaneum

为了克隆割手密SsWRKY1基因,并对其生物信息学和表达模式进行分析。本研究通过转录组测序和RT-PCR方法相结合首次从割手密中克隆到一个WRKY基因,利用生物信息学工具对其进行分析,并利用荧光定量PCR分析其在干旱胁迫下的表达模式。从割手密中克隆到一个全长1083 bp编码360个氨基酸的WRKY转录因子的基因,含有一个WRKY基因保守结构域和一个Cx4-5Cx22-23HxH的锌指结构域,属于WRKY基因家族的第Ⅱ类成员,命名为SsWRKY1,GenBank登录号为MH687932。荧光定量分析表明,随着干旱胁迫时间的延长,其表达量呈持续上调的表达模式,说明该基因是干旱胁迫诱导型基因。SsWRKY1基因在割手密应答干旱胁迫等非生物胁迫中可能起重要的调控作用,为进一步研究割手密的抗旱机制和甘蔗抗旱新品种的选育提供了科学依据。

In order to isolate and analysis the bioinformatics and expression pattern of SsWRKY1 gene from S.spontaneum.WRKY gene was first obtained by transcriptome sequencing and reversed transcription-polymerase chain reaction(RT-PCR)from S.spontaneum and analyzed by bioinformatics tools.The expression pattern responses under drought stress were analyzed by qRT-PCR.WRKY gene designated as SsWRKY1(GenBank accession number:MH687932),which incluing an ORF of 1083 bp coding sequence and coding 360 amino acids contained a conserved WRKY domains and a zinc finger structures(C2 H2)belong to GroupⅡof WRKY family,was amplified from Saccharum spontaneum L.Realtime-PCR analysis revealed that the expression level of SsWRKY1 was up regulated under different drought conditions,indicating that the gene is a drought stress-inducible gene.In this study,a novel gene of drought-resistance was found in S.spontaneum L.,which was to develop the excellent resources of wild species,as well as to explore the drought resistance of these resources,so as to provide the basis for the selection of the parents of the fine and drought resistance in the future.