黑果枸杞LrDREB1基因克隆及其在非生物胁迫下的表达分析

Cloning of LrDREB1 Gene of Lycium ruthenicum and Its Expression Analysis under Abiotic Stress

DREB(Dehydrate responsive element binding factor)转录因子是植物非生物逆境适应中的关键调节因子,该类转录因子具有保守的AP2/EREBP结构域,在低温、高盐等非生物逆境胁迫下,可调控下游逆境应答基因的表达,对增强植物的抗逆能力有重要作用。黑果枸杞具有极强的耐干旱、盐碱能力,为研究黑果枸杞DREB类基因在低温、盐碱响应中的功能,本研究以强抗盐灌木黑果枸杞总RNA为模板,基于前期转录组测序拼接序列,利用RT-PCR方法克隆得到一条DREB基因。该基因含有一个1116 bp的开放阅读框,编码372个氨基酸。系统进化树分析显示,Lr DREB1基因属于DREB亚家族A2组成员,与拟南芥AtDREB2B、AtDREB2E具有较高的相似性。荧光定量PCR结果显示,Lr DREB1受盐胁迫、ABA胁迫和低温胁迫诱导表达,可能参与依赖ABA的信号转导途径,调节黑果枸杞抗盐响应。本研究通过对黑果枸杞LrDREB1基因的克隆、序列分析和表达分析,为研究Lr DREB1的功能及黑果枸杞抗盐机理提供了帮助。

Dehydrate-responsive element binding(DREB)transcription factors is a key regulator in plant abiotic stress adaptation,with a typical AP2/EREBP DNA-binding domain.It can improve tolerance of plants by regulating numerous stress-responsive genes downstream when plants were suffered with abiotic stresses,including low temperature and high salt.Lycium ruthenicum is a shrub with extremely high salt resistance ability.In order to study the functions of DREB gene in the low temperature and saline-alkali response,we cloned a DREB gene by RT-PCR based on the previous transcriptome sequencing database from Lycium ruthenicum using total RNA as a template in this study.This gene had a 1116 bp open reading frame encoding 372 amino acids.Phylogenetic tree analysis showed that the Lr DREB1 gene belongs to the A2 group of the DREB subfamily and has high similarity to AtDREB2 B and AtDREB2 E in Arabidopsis thaliana.The results of real-time PCR showed that LrDREB1 was induced by salt,ABA and low temperature stresses,which indicated LrDREB1 participated in ABA-dependent signal transduction pathways and regulating the salt-and coldtolerant response of L.ruthenicum.In this paper,the cloning,sequence and expression analyses will be helpful for the foundation of the functional characterization of LrDREB1 involved in abiotic stresses response,especially salt-tolerance response of L.ruthenicum.