籽粒苋丙酮酸磷酸双激酶(PPDK)基因的克隆及其特征分析  

Cloning and Characterization of PPDK Gene from Amaranthus hypochondriacusne

作　　者：冯瑞云[1] 王原媛 闫建俊[1] 雷梦林[3] 郝雅萍[1] 白云凤[1] Feng Ruiyun;Wang Huijie;Yan Jianjun;Lei Menglin;Hao Yaping;Bai Yunfeng(Institute of Crop Sciences,Shanxi Academy of Agricultural Sciences,Key Laboratory of Crop Genetics and Molecular Improvement of Shanxi Province,Taiyuan,030031;Sunshine Union Hospital,Weifang,261061;Institute of Crop Germplasm Resources,Shanxi Academy of Agricultural Sciences,China Key Laboratory of Loess Plateau Crop Gene Resources and Germplasm Creation,Ministry of Agriculture,Taiyuan,030031)

机构地区：[1]山西省农业科学院作物科学研究所,作物遗传与分子改良山西省重点实验室,太原030031 [2]阳光融和医院,潍坊261072 [3]山西省农业科学院农作物品种资源研究所,农业部黄土高原作物基因资源与种质创制重点实验室,太原030001

出　　处：《基因组学与应用生物学》2019年第9期4120-4128,共9页Genomics and Applied Biology

基　　金：山西省主要农作物种质创新与分子育种山西省重点科技创新平台(201605D151002);山西省自然科学基金项目(201601D011075);山西省青年科技研究基金项目(201701D221210)共同资助

摘　　要：丙酮酸磷酸双激酶(pyruvate orthophosphate dikinase, PPDK)是C4植物在光合过程中的关键限速酶之一。本研究采用RT-PCR 技术,从籽粒苋叶片中克隆到籽粒苋丙酮酸磷酸双激酶基因cDNA全长。核苷酸序列分析表明,该基因cDNA序列开放阅读框全长为2 871 bp (GenBank 登录号: JF907701.1),可编码956 个氨基酸,分子量为104 kD。籽粒苋丙酮酸磷酸双激酶PPDK 与长寿花(Kalanchoe fedtschenko)、冰叶日中花(M.crystallinum)和板栗(Castanea mollissima)等双子叶植物PPDK 的同源性较高,分别为82.5%、82.0%和81.4%;与稗草(Echinochloa crusgalli)、高粱(Sorghum bicolor)和玉米(Zea mays)等单子叶植物PPDK 的序列一致性分别为74.5%、74.5%和73.9%。进化树分析表明,其与禾本科植物长寿花最为接近。半定量RT-PCR 研究表明,该基因受光诱导而上调表达,且在绿色叶片中的表达水平最高。克隆籽粒苋PPDK 基因将为今后作物高光效分子育种提供备选基因。Pyruvate phosphate dikinase (PPDK) is one of the speed-limiting enzymes in the photosynthetic process of C4 plants. A full-length cDNA of PPDK was isolated from Amaranthus hypochondriacus L. by RT-PCR technology. Nucleotides sequencing analysis showed that the open reading frame (ORF) of the gene including 2 871 bp, which encode 948 amino acids with the molecular weight of 104 kD. The PPDK of Amaranthus hypochondriacus L. has high homology to Kalanchoefedtschenko, M. crystallinum and Castaneamollissima, were about 82.5%, 82.0% and 81.4%, respectively;and that from endogen Echinochloacrusgalli, Sorghum bicolor and Zea mays were about 74.5%, 74.5% and 73.9%, respectively. The evolutionary tree analysis revealed that PPDK from Amaranthus hypochondriacus L. had a close genetic distance with Kalanchoefedtschenko. Semi-quantitative RT-PCR analysis showed that the expression of PPDK was significantly induced by light and it was highest expressed in leaf tissures. Cloning of the PPDK gene from Amaranthus hypochondriacus L. provided an important candidate gene for further molecular breeding.

关 键 词：籽粒苋 丙酮酸磷酸双激酶 克隆 表达 

分 类 号：Q78[生物学—分子生物学]