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园艺学报 ›› 2009, Vol. 36 ›› Issue (9): 1357-1380.

• 研究报告 • 上一篇    下一篇

6 - 磷酸葡萄糖酸脱氢酶基因(6PGDH) 的克隆与甜瓜属异源四倍体的分子验证

魏跃1,2;吴志明1;张蜀宁1;陈劲枫1*   

  1. (1南京农业大学, 南方蔬菜遗传改良重点开放实验室, 南京210095; 2 江苏农林职业技术学院, 江苏镇江212400)
  • 收稿日期:2009-03-13 修回日期:2009-07-14 出版日期:2009-09-25 发布日期:2009-09-25
  • 通讯作者: 陈劲枫

Cloning of 6-phosphoglucona te Dehydrogenase Gene (6PGDH) and Molecular Confirmation of Allotetraploid in Cucumis

WEI Yue1,2;WU Zhi-ming1;ZHANG Shu-ning1;CHEN Jin-feng1*   

  1. (1 Key Laboratory of Southern Vegetable Crop Genetics Improvement, Nanjing Agricultural University, Nanjing 210095, China;2 Polytechnical College of Agriculture and Forestry, Zhenjiang,Jiangsu 212400, China)
  • Received:2009-03-13 Revised:2009-07-14 Online:2009-09-25 Published:2009-09-25
  • Contact: CHEN Jin-feng

摘要: 根据已发表的黄瓜6 - 磷酸葡萄糖酸脱氢酶(6-phosphogluconate dehydrogenase ) 基因6PGDH的cDNA序列(GenBank登录号: EU815934) 设计引物, 扩增了甜瓜属异源四倍体新种及野生种和栽培黄瓜‘北京截头’的6PGDH基因片段, 测序结果表明3个种的6PGDH基因片段序列表现出高度的一致性,长度均为1 098 bp包含936 bp编码311个氨基酸的阅读框和162 bp的3′非翻译区末端, 片段内无内含子存在。利用DNAMAN软件检测到异源四倍体新种与野生种和栽培黄瓜‘北京截头’氨基酸的差异位点有3个, 碱基差异位点有22个, 差异位点的遗传分布符合孟德尔遗传学定律, 从而在分子水平证实了异源四倍体新种是野生种和栽培黄瓜‘北京截头’的杂交后代。

关键词: 黄瓜, 异源四倍体, 6 - 磷酸葡萄糖酸脱氢酶, 杂交种, 克隆

Abstract: In order to demonstrate that the synthetic allotetrop loid species (Cucum is hytivus Chen and Kirkbride. , 2n = 38) was the hybrids progeny originated from crossing between the wild Cucumis species (C.hystrix Chakr. , 2n = 24) and the cultivated cucumber‘Beijing J ietou’(C. sa tivus L. , 2n = 14) in molecular level, primers were designed according to the 6-phosphogluconate dehydrogenase ( 6PGDH) gene cDNA sequence of cucumber (GenBank accession number: EU815934). The 6PGDH gene fragmentswere amplified
from allotetroploid, the wild Cucumis and cultivated cucumber‘Beijing Jietou’, respectively. Sequence analysis showed that the 6PGDH fragments in three species were highly homologous, having 1 098 bp length each.It encompassed of 936 bp long open reading frame (ORF) encoding 311 amino acids and had 162 bp non-translation 3′- terminal end region (3′UTR) , no intron existed in 6PGDH fragment.The sequences were analyzed by software program DNAMAN, detected three variable amino acid locips and twenty-two bases locips.The inheritance of variable locips were congruence with theMendelps genetics law of inheritance, therefore, it is proven atmolecular level that this synthetic allotetraploid species was the hybrid originated from C. hystrix and C. sativus‘Beijing J ietou’crossing.

Key words: cucumber, allotetrop loid, 6PGDH, hybrid, clone

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