中国农业科学 ›› 2010, Vol. 43 ›› Issue (22): 4663-4670 .doi: 10.3864/j.issn.0578-1752.2010.22.013

• 园艺 • 上一篇    下一篇

黄瓜中导入野生酸黄瓜外源DNA片段的分子验证

史建磊,陈劲枫,娄群峰,隋益虎,吴志明,张淑霞

  

  1. (南京农业大学园艺学院/作物遗传与种质创新国家重点实验室)
  • 收稿日期:2010-03-19 修回日期:2010-06-07 出版日期:2010-11-15 发布日期:2010-11-15
  • 通讯作者: 陈劲枫

Molecular Verification of DNA Flow from Wild Sour Cucumber to Cultivated Cucumber

SHI Jian-lei, CHEN Jin-feng, LOU Qun-feng, SUI Yi-hu, WU Zhi-ming, ZHANG Shu-xia
  

  1. (南京农业大学园艺学院/作物遗传与种质创新国家重点实验室)
  • Received:2010-03-19 Revised:2010-06-07 Online:2010-11-15 Published:2010-11-15
  • Contact: CHEN Jin-feng

摘要:

【目的】对栽培黄瓜(Cucumis sativus L.,2n=14)与野生酸黄瓜(Cucumis hystrix Chakr.,2n=24)经远缘杂交和多代回交、自交而来的渐渗系进行分子标记分析和表型鉴定,旨在为黄瓜导入酸黄瓜外源DNA片段提供相应依据。【方法】从分子标记分析、DNA序列比对和农艺性状表现上对各渐渗系和双亲进行研究。【结果】渐渗系与栽培黄瓜存在13.2%的遗传多态性,包括供体特异带、缺失带和产生的新带。利用RAPD引物D-11和SSR引物06632在渐渗系中分别扩增出约310 bp和150 bp的酸黄瓜特异带,序列分析表明其与酸黄瓜相应DNA片段的相似性分别为92.93%和96.48%,碱基差异分别为41个和9个,包括碱基转换、颠换和缺失。表型鉴定发现其呈现出酸黄瓜的遗传特性。【结论】野生酸黄瓜DNA已渐渗进入栽培黄瓜基因组,同时发生了少量不同类型的变异。

关键词: 黄瓜, 野生酸黄瓜, 渐渗系, DNA序列, 农艺性状

Abstract:

【Objective】 Introgression lines were researched by molecular markers and the phenotypic identification in order to proof DNA flow from wild sour cucumber to cultivated cucumber, which were obtained by interspecific hybridization between the cultivated cucumber (Cucumis sativus L., 2n=14) and wild sour cucumber (Cucumis hystrix Chakr., 2n=24), and multiple generational backcrossing and selfing followed. 【Method】 Molecular markers, DNA sequences and agronomic traits were used in the study to analyze and contrast introgression lines and their parents. 【Result】 The results showed that the genetic polymorphism between introgression lines and cultivated cucumber was 13.2%, including donor specific bands, missing bands and new bands. The sour cucumber specific bands in introgression lines were obtained by RAPD primer D-11 and SSR primer 06632, with the size of 310 bp and 150 bp respectively. Sequence analysis showed that the identity was 92.93% and 96.48%, respectively, though there were 41 and 9 different base pairs, respectively, including the base pairs transition, transversion and deletion. And the genetic characteristics of sour cucumber were found in introgression lines. 【Conclusion】 These results confirmed that a few mutations of different types occurred with DNA segments of wild sour cucumber being introgressed into cultivated cucumber.

Key words: Cucumis sativus L., wild sour cucumber, introgression lines, DNA sequence, agronomic traits