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园艺学报 ›› 2011, Vol. 38 ›› Issue (6): 1111-1120.

• 蔬菜 • 上一篇    下一篇

马铃薯GLDH基因的克隆及序列分析

董玉梅1,3,密其鹏1,焦自高3,杨元军3,于贤昌2,*   

  1. (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,山东泰安 271018;2中国农业科学院蔬菜花卉研究所,北京 100081;3山东省农业科学院蔬菜研究所,济南 250100)   
  • 收稿日期:2011-01-28 修回日期:2011-04-25 出版日期:2011-06-25 发布日期:2011-06-25
  • 通讯作者: 于贤昌

Cloning and Sequence Analysis of L-galactono-1,4-lactone Dehydrogenase Gene(GLDH)from Solanum tuberosum

DONG Yu-mei1,3,MI Qi-peng1,JIAO Zi-gao3,YANG Yuan-jun3,and YU Xian-chang2,*   

  1. (1State Key Laboratory of Crop Biology,College of Horticulture Science and Engineering,Shandong Agricultural University,Tai’an,Shandong 271018,China;2 The Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China;3Vegetable Research Institute,Shandong Academy of Agricultural Sciences,Ji’nan 250100,China)   
  • Received:2011-01-28 Revised:2011-04-25 Online:2011-06-25 Published:2011-06-25
  • Contact: YU Xian-chang

摘要: 以马铃薯品种‘Favorita’叶片中的cDNA为模板,采用RT-PCR、巢式PCR、3′RACE和5′RACE技术,获得了L–半乳糖酸–1,4–内酯脱氢酶(EC 1131213,GLDH)基因cDNA 2 563 bp的全长序列,命名为StGLDH(GenBank:FJ755844)。序列分析表明,该基因编码区长1 773 bp,编码590个氨基酸,与其他植物GLDH氨基酸序列具有很高的同源性,尤其与番茄、辣椒、烟草GLDH 氨基酸序列具有90.6% ~ 95.9%的同源性。荧光定量分析表明,该基因在马铃薯不同器官中均有表达,在幼叶和功能叶中表达量最高,在茎中表达量最低;除匍匐茎外,其它器官中抗坏血酸(ascorbic acid,AsA)含量与StGLDH的表达高度一致。  

关键词: 马铃薯, L–抗坏血酸, L–半乳糖酸–1, 4–内酯脱氢酶, 克隆, 基因表达

Abstract: A full-length cDNA clone encoding L-galactono-1,4-lactone dehydrogenase(GLDH),named as StGLDH(GenBank:FJ755844),was isolated from the leaf of potato(Solanum tuberosum L.)cv. Favorita by RT-PCR,nested PCR and RACE. StGLDH transcript is 2 563 bp long with an open reading frame of 1 773 bp and encodes a polypeptide of 590 amino acids. Sequence analysis showed that deduced StGLDH protein was highly homologous to other GLDH proteins from different species,especially with Solanum lycopersicum,Capsicum annuum,Nicotiana tabacum,about 90.6%–95.9% identity in amino acid sequence.Real time PCR analysis indicated that StGLDH expressed in different organs. The transcription level of StGLDH in mature leaves and young leaves were higher than that in old leaves,stems,roots and tubers respectively. The ascorbic acid content was positively related to the StGLDH transcription level in all organs except the ascorbic acid content in stolon.  

Key words: Solanum tuberosum L., ascorbic acid, L-galactono-1, 4-lactone dehydrogenase(GLDH), cloning, gene expression

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