华北农学报 ›› 2016, Vol. 31 ›› Issue (4): 94-99. doi: 10.7668/hbnxb.2016.04.016

所属专题: 抗旱节水

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拟南芥CPK10/CPK30双突变体的构建及表型分析

谢鑫, 孙宁, 魏凤菊   

  1. 河北农业大学 生命科学学院, 河北省植物生理与分子病理学重点实验室, 河北 保定 071001
  • 收稿日期:2016-05-18 出版日期:2016-08-28
  • 通讯作者: 魏凤菊(1979-),女,河北衡水人,讲师,博士,主要从事植物抵抗逆境分子机制方面的研究。
  • 作者简介:谢鑫(1990-),女,河北邯郸人,在读硕士,主要从事植物逆境研究。
  • 基金资助:
    国家自然科学基金专项(31040052);国家自然科学基金青年基金项目(31101022);植物生理学与生物化学国家重点实验室开放课题项目(SKLPPBKF1504)

Construction and Phenotype Analysis of CPK10/CPK30 Double Mutantin Arabidopsis

XIE Xin, SUN Ning, WEI Fengju   

  1. College of Life Sciences, Agricultural University of Hebei, Hebei Province Key Laboratory of Plant Physiology and Molecular Pathology, Baoding 071001, China
  • Received:2016-05-18 Published:2016-08-28

摘要: CDPKs是植物细胞内一类重要的钙感受器,拟南芥CPK10属于CDPK家族成员。为研究CPK10及与其同源性较高的CPK30是否共同参与逆境响应。首先构建了cpk10×cpk30双突变体,然后进行多种逆境下的生理表现检测,并利用RT-PCR方法分析2个基因的表达情况。结果显示,模拟干旱、盐、ABA处理拟南芥幼苗后,双突变体与野生型无差异;成苗期双突变体与单突变体对干旱敏感程度类似。转录水平检测到,在干旱胁迫时双突变体中RD29A的表达与野生型及单突变体趋势相反,呈下降趋势;响应ABA的OST1在双突变体中受干旱诱导后0.5 h明显表达上调。成功获得了cpk10×cpk30双突变体,由生理表型和表达分析结果推测CPK10CPK30可能共同参与了依赖ABA的干旱逆境信号转导过程,且二者之间存在功能冗余。

关键词: 钙依赖的蛋白激酶, 双突变体, 逆境胁迫, 表型分析

Abstract: Calcium-dependent protein kinases (CDPKs) function as Ca2+ sensor in various plant physiological processes.CPK10 was one member of CDPK family.In order to investigate the molecular mechanisms of CPK10 and it's high homology CPK30 in response to stress,cpk10×cpk30 double mutant was firstly constructed.The phenotypes in response to various stress were detected,and the RT-PCR method was used to analyzed the two genes expression.The results showed that there was no difference in the seed germination stage between double mutant and wild-type plants in the treatment of drought,salt and ABA.In the mature stage,double mutant exhibited no difference to drought stress compared with wild-type and single mutant.Nevertheless,the expression levels of RD29A was decreased in the double mutant and showed opposite trend in wild-type and single mutant during drought stress.The expression of ABA-responsive gene OST1 was enhanced obviously in double mutant after drought stress 0.5 h.The cpk10×cpk30 double mutant was successfully obtained.Our data indicated that CPK10 and CPK30 might be involved in ABA-dependent signal transduction pathway,and showed redundant functions.

Key words: CDPK, Double mutant, Adversity stress, Phenotype analysis

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引用本文

谢鑫, 孙宁, 魏凤菊. 拟南芥CPK10/CPK30双突变体的构建及表型分析[J]. 华北农学报, 2016, 31(4): 94-99. doi: 10.7668/hbnxb.2016.04.016.

XIE Xin, SUN Ning, WEI Fengju. Construction and Phenotype Analysis of CPK10/CPK30 Double Mutantin Arabidopsis[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(4): 94-99. doi: 10.7668/hbnxb.2016.04.016.

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