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  

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

作　　者：魏跃[1,2] 吴志明[1] 张蜀宁[1] 陈劲枫[1] 

机构地区：[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

出　　处：《园艺学报》2009年第9期1375-1380,共6页Acta Horticulturae Sinica

基　　金：国家自然科学基金重点项目(30830079);国家自然科学基金项目(30671419;30700541);国家高技术研究发展计划专项(2008AA10Z150;2006AA10Z1A8;2006AA100108);国家重点基础研究发展计划项目(2009CB119000);国家科技支撑计划项目(2008BADB105;2006BAD13B06;2006BAD01A7-5-11)

摘　　要：根据已发表的黄瓜6-磷酸葡萄糖酸脱氢酶(6-phosphog luconate 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1098bp包含936bp编码311个氨基酸的阅读框和162bp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。In order to demonstrate that the synthetic allotetroploid species （ Cucumis 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 Jietou＇ （C. sativus 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 fragments were 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 loci＇s and twenty-two bases loci＇s. The inheritance of variable loci＇s were congruence with the Mendel＇s genetics law of inheritance, therefore, it is proven at molecular level that this synthetic allotetraploid species was the hybrid originated from C. hystrix and C. sativus ＇ Beijing Jietou＇ crossing.

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 

分 类 号：S652[农业科学—果树学]