Acta Agrestia Sinica ›› 2022, Vol. 30 ›› Issue (7): 1659-1667.DOI: 10.11733/j.issn.1007-0435.2022.07.006

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Cloning and Abiotic Stress Response Analysis of SnRK2.2 Gene in Kentucky Bluegrass

JIN Yi-feng1,2, CHEN Yang1,2, GAO Yan-song1,2, YOU Xue1, XIONG Yi1,2, ZHAO Qing-feng1, LAN Hong-yu3   

  1. 1. College of Life Science and Agro-Forestry, Qiqihar Univesity, Qiqihar, Heilongjiang Province 161006, China;
    2. Heilongjiang Province Key Laboratory of Resistance Gene Engineering and Preservation of Biodiversity in Cold Areas, Qiqihar, Heilongjiang Province 161006, China;
    3. Qiqihar Branch, Heilongjiang Academy of Agricultural Sciences, Qiqihar, Heilongjiang Province 161006, China
  • Received:2022-01-04 Revised:2022-02-18 Published:2022-08-02

草地早熟禾SnRK2.2基因克隆及非生物胁迫响应分析

金一锋1,2, 陈阳1,2, 高岩松1,2, 尤学1, 熊毅1,2, 赵清峰1, 兰红宇3   

  1. 1. 齐齐哈尔大学生命科学与农林学院, 黑龙江 齐齐哈尔 161006;
    2. 抗性基因工程与寒地生物多样性保护黑龙江省重点实验室, 黑龙江 齐齐哈尔 161006;
    3. 黑龙江省农业科学院齐齐哈尔分院, 黑龙江 齐齐哈尔 161006
  • 通讯作者: 陈阳,E-mail:Chenyang8368215@126.com
  • 作者简介:金一锋(1984-),男,朝鲜族,黑龙江鹤岗人,讲师,博士研究生,主要从事草坪草逆境分子生物学研究,E-mail:Jinyifeng8368215@126.com
  • 基金资助:
    国家自然科学基金(31701958);黑龙江省普通高等学校青年创新人才培养计划项目(UNPYSCT-2018102);黑龙江省省属高等学校基本科研业务费科研项目(145109518);齐齐哈尔大学大学生创新创业项目(202110232361、202110232371、202110232392)资助

Abstract: Protein kinases are essential regulatory factors in plants. Sucrose non-fermenting 1-related protein kinase 2 (SnRK2) plays a crucial role in the plant stress signal transduction pathway through phosphorylation during adversity stress,and is participated in the interaction between metabolism and stress signals. To explore and utilize the SnRK2 family gene of Poa pratensis L,in this study,the SnRK2.2 gene of Poa Pratensis L was cloned by RT-PCR based on the annotation information of the transcriptome data. Bioinformatics and real-time quantitative PCR were utilized to investigate its molecular characteristics,tissue location and expression of the gene under abiotic stress. The results showed that the SnRK2 gene of Poa Pratensis L contains typical STKc_SnRK2 domain,ATP binding signal region of protein kinase,N-myristoylation site,active site of serine/threonine-protein kinase and other functional domains,and it has the highest homology with oat and wheat. The results of real-time fluorescent quantitative PCR showed that the relative expression of the SnRK2.2 gene in tissues was panicle > leaf > stem > root. The SnRK2 gene of Poa Pratensis L could respond to drought,salt,low nitrogen and low phosphorus stress. The results provide a theoretical basis for enriching the SnRK2 resistance mechanism in turfgrass.

Key words: Kentucky Bluegrass, Protein Kinase, SnRK2.2, Gene Cloning, Expression Analysis

摘要: 逆境胁迫期间蔗糖非发酵相关的蛋白激酶2(SnRK2)通过磷酸化在植物胁迫信号转导途径中起关键作用,参与代谢和应激信号之间的相互作用。为发掘并利用草地早熟禾(Poa pratensis L.) SnRK2家族基因,本研究利用RT-PCR技术得到SnRK2.2基因,借助生物信息学及实时荧光定量PCR技术分别对其进行分子特征、组织部位以及非生物胁迫下该基因表达模式的研究。研究结果:草地早熟禾SnRK2.2包含典型STKc_SnRK2结构域、蛋白激酶ATP结合信号区、N-肉豆蔻酰化位点等功能域,其与燕麦、小麦同源性最高;实时荧光定量PCR结果表明,草地早熟禾SnRK2.2基因在组织部位中相对表达量为穗>叶>茎>根,该基因可积极响应干旱、盐、低氮、低磷、ABA、BR诱导处理。研究结果为丰富草坪草SnRK2抗逆机制提供理论基础。

关键词: 草地早熟禾, 蛋白激酶, SnRK2.2, 基因克隆, 表达分析

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