梅花2个PmWRKY2基因克隆及在逆境胁迫下的表达模式

Cloning and expression analysis under adversity stress of 2 PmWRKY2 in Prunus mume

【目的】低温是影响梅花Prunus mume栽培应用的重要环境因素。WRKY基因是一类主要存在于植物中的转录因子,参与响应非生物胁迫等过程。了解梅花WRKY基因对非生物和脱落酸(ABA)胁迫的响应,对梅花定向育种具有重要的意义。【方法】以梅花‘骨红朱砂’Prunus mume‘Guhong Zhusha’为材料,通过反转录PCR(RT-PCR)克隆获得了2个WRKY2转录因子,命名为PmWRKY2-1和PmWRKY2-2;采用实时荧光定量PCR(qRT-PCR)分析PmWRKY2-1和PmWRKY2-2基因在低温和干旱下的表达模式。【结果】PmWRKY2-1和PmWRKY2-2的编码区长度分别为2223和2220 bp,分别编码740和739个氨基酸,包含2个WRKY结构域和C2H2型锌指结构;PmWRKY2-1与PmWRKY2-2亲缘关系较远,但两者与蔷薇科Rosaceae植物欧洲甜樱桃P.avium、桃P.persica、李P.dulcis的亲缘关系较近。qRT-PCR结果显示:在低温和干旱处理下,PmWRKY2-1与PmWRKY2-2都能被诱导;脱落酸(ABA)处理后,PmWRKY2-1与PmWRKY2-2的表达显著降低。【结论】PmWRKY2-1与PmWRKY2-2可能参与调控梅花低温和干旱响应,并可能受到ABA的调控。图6表1参28。

[Objective]Low temperature is a main environmental factor that influences the cultivation and application of Prunus mume whereas WRKY gene is a plant-specific transcription factor which participates in the response to abiotic stress process.This study,with an investigation of how WRKY gene responds to low temperature and drought stress,is aimed to provide guidance for the directional breeding of P.mume.[Method]With the P.mume‘Guhong Zhusha’cDNA template selected as the substance,two WRKY2 genes were cloned by means of RT-PCR,named as Pm WRKY2-1 and Pm WRKY2-2 before their expression patterns were under low temperature and in the condition of drought employing real-time quantitative PCR(qRT-PCR).[Result]a)Pm WRKY2-1 and Pm WRKY2-2,with respective coding area lengths of 2223 and 2220 bp,encode740 and 739 amino acids respectively,both including 2 WRKY domains and a C2H2 zinc finger structure;b)though with a distant genetic relationshp with each other,both Pm WRKY2-1 and Pm WRKY2-2 had a close relationship with P.avium,P.persica and P.dulcis;c)according to the results of the real-time quantitative PCR(qRT-PCR),both Pm WRKY2-1 and Pm WRKY2-2 could be induced by low temperature and drought treatment And d)the expressions of Pm WRKY2-1 and Pm WRKY2-2 were significantly reduced after abscisic acid(ABA)treatment.[Conclusion]Pm WRKY2-1 and Pm WRKY2-2 are likely to participate in the regulation of low temperature and drought response of P.mume,yet might be subject to the regulation by ABA.[Ch,6 fig.1 tab.28 ref.]