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园艺学报 ›› 2018, Vol. 45 ›› Issue (7): 1225-1236.doi: 10.16420/j.issn.0513-353x.2017-0726

• 研究论文 •    下一篇

苹果蛋白磷酸酶基因MdPP2C-Like的克隆及其对ABA的响应分析

张春玲,周李杰,王桂栾,李媛媛,王小非*,郝玉金*   

  1. (山东农业大学园艺科学与工程学院,作物生物学国家重点实验室,国家苹果工程技术研究中心,山东泰安 271018)
  • 出版日期:2018-07-25 发布日期:2018-07-25

Molecular Cloning of Protein Phosphatase Gene MdPP2C-Like in Apple and the Response Analysis of ABA

ZHANG Chunling,ZHOU Lijie,WANG Guiluan,LI Yuanyuan,WANG Xiaofei*,and HAO Yujin*   

  1. (College of Horticulture Science and Engineering,Shandong Agricultural University,State Key Laboratory of Crop Biology,National Research Center for Apple Engineering and Technology,Tai’an,Shandong 271018,China)
  • Online:2018-07-25 Published:2018-07-25

摘要:

以‘嘎拉’苹果(Malus × domestica Borkh.)为试材,克隆了ABA信号转导过程中的响应基因蛋白磷酸酶基因MdPP2C-Like(基因序列号 MDP0000126636)。基因结构分析显示,MdPP2C-Like定位于苹果8号染色体上,含有5个外显子与4个内含子;该基因全长1 035 bp,编码344个氨基酸,预测其蛋白质分子量为37.84 kD,等电点(pI)为5.72。蛋白质结构分析发现,其存在1个蛋白磷酸酶2C(PP2C)保守结构域,并与拟南芥中的A类PP2C保守结构域高度同源。构建的系统进化树表明MdPP2C-Like与At4G32950位于同一进化支,同源性较高。亚细胞定位结果表明,MdPP2C-Like主要定位于细胞核,少数分布在细胞质。定量分析显示,MdPP2C-Like在苹果不同组织中表达量不同,根中表达量较高。对获得的转基因拟南芥株系和苹果愈伤组织进行ABA响应分析,结果表明该基因的过量表达降低了植株和愈伤组织对ABA的敏感性。以上结果表明,克隆得到的MdPP2C-Like在响应ABA信号过程中起着重要作用。

关键词: 苹果, MdPP2C-Like, 蛋白磷酸酶, 过表达, ABA

Abstract:

Protein phosphatase gene MdPP2C-Like(MDP0000126636),an important factor that was involved in ABA signal pathway,was cloned from apple(Malus × domestica Borkh.). Gene structure analysis showed that MdPP2C-Like gene contained 5 exons and 4 introns,and it was located on the eighth chromosome of apple genome. After amplification,a 1 035 bp open reading frame(ORF)was obtained. MdPP2C-Like gene encoded a protein of 344 amino acids with a calculated mass of 37.84 kD and an isoelectric point(pI)of 5.72. MdPP2C-Like protein sequence comprised a conserved phosphatase 2C (PP2C)domain,just as group A AtPP2Cs. The phylogenetic tree of the construction indicated that MdPP2C-Like and At4G32950 were located in the same evolutionary branch and had high homology. MdPP2C-Like gene mainly localized in the nucleus,with a few in cytoplasm. The expression level of MdPP2C-Like was different in all apple tissues,with the highest level in roots. Moreover,overexpression of MdPP2C-Like gene in apple callus and Arabidopsis plants reduced the sensitivity to ABA. Taken together,these findings showed that MdPP2C-Like played an important role in response to ABA signaling.

Key words: apple, MdPP2C-Like, protein phosphatase, overexpression, ABA

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