草地早熟禾蛋白激酶SnRK2.4基因克隆及非生物胁迫响应分析  

Cloning of SnRK2.4 Gene and It′s Response to Abiotic Stress in Kentucky Bluegrass

作　　者：金一锋[1,2] 高岩松 王琦 王萌萌 赵迪 熊毅 陈阳[1,2] JIN Yifeng;GAO Yansong;WANG Qi;WANG Mengmeng;ZHAO Di;XIONG Yi;CHEN Yang(College of Life Science and Agro-Forestry,Qiqihar Univesity,Qiqihar 161006,China;Heilongjiang Province Key Laboratory of Resistance Gene Engineering and Preservation of Biodiversity in Cold Areas,Qiqihar 161006,China)

机构地区：[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

出　　处：《华北农学报》2022年第5期9-15,共7页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31701958);黑龙江省自然科学基金项目(QC2017026);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(135309365);齐齐哈尔大学大学生创新创业项目(202110232361,202110232371,202110232392)。

摘　　要：蛋白激酶是植物逆境防御机制中重要的核心成分,植物蛋白激酶SnRK2是一种丝氨酸/苏氨酸蛋白激酶,可通过磷酸化途径在植物逆境胁迫响应中发挥重要作用。分析草地早熟禾蛋白激酶基因SnRK2.4在非生物胁迫下的表达规律,旨在揭示其在逆境调控中的作用,以优质的冷季型草坪草草地早熟禾为试材,利用RT-PCR方法克隆得到SnRK2.4基因,其包含一个1092 bp开放阅读框区,编码363个氨基酸序列,利用生物信息学分析其分子特征,并采用实时荧光定量PCR方法观测不同组织部位、非生物胁迫下该基因表达模式。结果表明,草地早熟禾SnRK2.4属于SRK/SAPK家族,包含典型的STKc_SnRK2结构域,酪氨酸激酶磷酸化位点、酪蛋白激酶Ⅱ磷酸化位点、丝氨酸/苏氨酸蛋白激酶活性位点等功能域,其与二穗短柄草同源性最高。实时荧光定量PCR结果表明,草地早熟禾SnRK2.4基因表达水平存在组织特异性,穗部表达量最高,根、茎、叶中无显著差异,草地早熟禾SnRK2.4基因积极响应干旱、盐、低氮、低磷、ABA、BR等非生物胁迫。Protein kinases are important factors in plant defense system,protein kinase SnRK2 is a serine/threonine protein kinase,can play an important role in the plant stress signal transduction pathway through phosphorylation.We analyzed the expression pattern of SnRK2.4 under abiotic stresses,aiming to reveal its role in the regulation of adversity.The SnRK2.4 gene of high-quality cold-season turfgrass Poa pratensis L.was cloned using RT-PCR,the SnRK2.4 gene contained an ORF of 1092 bp encoding a 363-amino acid protein,and its molecular characteristics were analyzed by bioinformatics.In addition,the expression pattern of this gene in different tissue parts under different abiotic stresses was observed using qPCR.The results showed that the Poa pratensis L.SnRK2.4 gene belonged to the SRK/SAPK superfamily,contained typical STKc_SnRK2 domain,tyrosine kinase phosphorylation site,casein kinaseⅡphosphorylation site and serine/threonine-protein kinase activity site,which had the highest homology with Brachypodium distachyon.The qPCR results demonstrated that the Poa pratensis L.SnRK2.4 gene was tissue-specific,highly expressed in panicles,and had no significant differential expression in roots,stems and leaves.Moreover,the Poa pratensis L.SnRK2.4 gene could respond positively to abiotic stresses such as drought,salt,low nitrogen,low phosphorus,ABA and BR.

关 键 词：草地早熟禾 蛋白激酶 SnRK2.4 基因克隆 非生物胁迫 

分 类 号：Q78[生物学—分子生物学]