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苹果蛋白磷酸酶基因MdPP2C-Like的克隆及其对ABA的响应分析

苹果蛋白磷酸酶基因MdPP2C-Like的克隆及其对ABA的响应分析

ISSN:0513-353X
2018年第45卷第7期
研究论文
张春玲,周李杰,王桂栾,李媛媛,王小非,郝玉金 ZHANG Chunling,ZHOU Lijie,WANG Guiluan,LI Yuanyuan,WANG Xiaofei,and HAO Yujin
(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) (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)

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。

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.

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ISSN:0513-353X
2018年第45卷第7期
研究论文

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