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梅花2个PmWRKY2基因克隆及其在逆境胁迫下的表达模式

王楠楠 董彬 杨丽媛 赵宏波

王楠楠, 董彬, 杨丽媛, 赵宏波. 梅花2个PmWRKY2基因克隆及其在逆境胁迫下的表达模式[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20200706
引用本文: 王楠楠, 董彬, 杨丽媛, 赵宏波. 梅花2个PmWRKY2基因克隆及其在逆境胁迫下的表达模式[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20200706
WANG Nannan, DONG Bin, YANG Liyuan, ZHAO Hongbo. Cloning and expression analysis under adversity stress of 2 PmWRKY2 in Prunus mume[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20200706
Citation: WANG Nannan, DONG Bin, YANG Liyuan, ZHAO Hongbo. Cloning and expression analysis under adversity stress of 2 PmWRKY2 in Prunus mume[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20200706

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梅花2个PmWRKY2基因克隆及其在逆境胁迫下的表达模式

doi: 10.11833/j.issn.2095-0756.20200706
基金项目: “十三五”国家重点研发计划资助项目(2019YFD1001505,2018YFD1000402);浙江省农业新品种选育重点项目(2016C02056-12)
详细信息
    作者简介: 王楠楠(ORCID: 0000-0002-2571-5980),从事抗寒分子机制研究。E-mail: 1780192738@qq.com
    通信作者: 赵宏波(ORCID: 0000-0003-4714-8240),教授,博士,博士生导师,从事观赏植物遗传育种等研究。E-mail: zhaohb@zafu.edu.cn
  • 中图分类号: S685.17

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

  • 摘要:   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2 223和2 220 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植物樱桃李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
  • 图  1  2个PmWRKY2基因的扩增

    Figure  1  PCR amplification of 2 PmWRKY2 genes in Prunus mume

    图  2  梅花与其他物种WRKY氨基酸序列的比对

    Figure  2  Amino acid sequence of WRKY between Prunus mume and other species

    图  3  PmWRKY2-1和PmWRKY2-2的WRKY结构域

    Figure  3  WRKY domain displays of PmWRKY2-1和PmWRKY2-2

    图  4  PmWRKY2-1和PmWRKY2-2蛋白质的二级结构

    Figure  4  Secondary protein structure of PmWRKY2-1 and PmWRKY2-2

    图  5  梅花与其他物种WRKY氨基酸序列系统进化树分析

    Figure  5  Phylogenetic tree analysis of the amino acid sequence of P. mume and other species

    图  6  非生物和ABA处理下的PmWRKY2-1和PmWRKY2-2的表达

    Figure  6  Expression levels of PmWRKY2-1 and PmWRKY2-2 under abiotic stress and ABA

    表  1  基因克隆及表达所用引物序列

    Table  1.   Primers used in Gene clone and Quantitative real-time PCR

    用途引物名称序列(5′→3′)
    基因克隆PmWRKY2-1FATGGCTGGCATCGATGA
    PmWRKY2-1RCTACATCTGTGGTCCAAG
    PmWRKY2-2FATGGGATTTTTAAGAACC
    PmWRKY2-2RCTAGTACGATTGATGACTGCTTC
    实时荧光定量PCRQPmWRKY2-1FGTCCCCTTATCTGACAATACCTC
    QPmWRKY2-1RAAAGCGAATGAAGTATTTATGTCCT
    QPmWRKY2-2FTCCGTTGCTTCCTCCCAATGATGAC
    QPmWRKY2-2RCAAAATCTATTGGTTGTTGCTCC
    QPmEF1αSCGGATTCAATGTTAAGAATGTTGC
    QPmEF1αAAGAACTGGAGCATATCCGTTACC
    下载: 导出CSV
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  • 收稿日期:  2020-11-11
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梅花2个PmWRKY2基因克隆及其在逆境胁迫下的表达模式

doi: 10.11833/j.issn.2095-0756.20200706
    基金项目:  “十三五”国家重点研发计划资助项目(2019YFD1001505,2018YFD1000402);浙江省农业新品种选育重点项目(2016C02056-12)
    作者简介:

    王楠楠(ORCID: 0000-0002-2571-5980),从事抗寒分子机制研究。E-mail: 1780192738@qq.com

    通信作者: 赵宏波(ORCID: 0000-0003-4714-8240),教授,博士,博士生导师,从事观赏植物遗传育种等研究。E-mail: zhaohb@zafu.edu.cn
  • 中图分类号: S685.17

摘要:   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2 223和2 220 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植物樱桃李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

English Abstract

王楠楠, 董彬, 杨丽媛, 赵宏波. 梅花2个PmWRKY2基因克隆及其在逆境胁迫下的表达模式[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20200706
引用本文: 王楠楠, 董彬, 杨丽媛, 赵宏波. 梅花2个PmWRKY2基因克隆及其在逆境胁迫下的表达模式[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20200706
WANG Nannan, DONG Bin, YANG Liyuan, ZHAO Hongbo. Cloning and expression analysis under adversity stress of 2 PmWRKY2 in Prunus mume[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20200706
Citation: WANG Nannan, DONG Bin, YANG Liyuan, ZHAO Hongbo. Cloning and expression analysis under adversity stress of 2 PmWRKY2 in Prunus mume[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20200706

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