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园艺学报 ›› 2015, Vol. 42 ›› Issue (2): 341-349.doi: 10.16420/j.issn.0513-353x.2014-0882

• 研究报告 • 上一篇    下一篇

刺梨转录组SSR信息分析及其分子标记开发

鄢秀芹,鲁敏,安华明   

  1. 贵州大学农学院,贵州省果树工程技术研究中心,贵阳 550025
  • 出版日期:2015-02-25 发布日期:2015-02-25
  • 基金资助:
    国家自然科学基金项目(31360475);贵州省重大科技专项项目(黔科重大专项字20136006-1);贵州大学研究生创新基金项目(研农2014004)

Analysis on SSR Information in Transcriptome and Development of Molecular Markers in Rosa roxburghii

YAN Xiu-qin,LU Min,and AN Hua-ming   

  1. College of Agriculture,Guizhou University,Guizhou Engineering Research Center for Fruit Crops,Guiyang 550025,China
  • Online:2015-02-25 Published:2015-02-25

摘要: 利用MISA软件筛选刺梨(Rosa roxburghii Tratt)转录组测序获得的106 590条Unigene,共检测出21 711个SSR位点,分布于18 155条Unigene中,出现频率为20.37%,平均分布距离为1.68 kb。优势重复基序为三核苷酸、四核苷酸和二核苷酸,分别占总SSR位点的26.87%、26.77%和24.93%。AG/CT与AAG/CTT分别是二核苷酸与三核苷酸的优势重复基元,分别占总SSR重复类型的17.80%和11.55%。以不同主导重复基元类型设计合成42对SSR引物,并以刺梨及其他蔷薇属种质的基因组DNA为模板对其有效性及通用性进行初步验证,筛选出具有清晰扩增产物的引物23对,并且均在同属材料中通用。选取16份贵州刺梨资源对筛选出的引物进行多态性检测,获得12对具有多态性的引物。以上结果表明,刺梨转录组测序产生的Unigene信息可作为开发SSR标记的有效来源,获得的大批量SSR标记可为刺梨及其近缘种的遗传多样性分析和遗传图谱构建提供更加丰富可靠的标记选择。

关键词: 刺梨, SSR, 转录组

Abstract: One hundred and six thousand five hundred and ninety unigenes from fruit transcriptome of Rosa roxburghii Tratt were screened using MISA software. A total of 21 711 SSRs that occurred in 18 155 unigenes were identified,and the frequency of these SSRs was 20.37% and mean distance was 1.68 kb in the unigenes. Trinucleotide,tetranucleotide and dinucleotide were major types,accounting for 26.87%,26.77% and 24.93%,respectively. AG/CT,AAG/CTT were respectively most frequent motifs in dinucleotide and trinucleotide repeats,accounting for 17.80% and 11.55%,respectively. Using the Primer 3.0,42 primers were designed and synthesized based on different dominant motifs types,and verified with R. roxburghii germplasms for validity and Rosa germplasms for transferability. The results showed that the products of 23 primers are clear and effective,23 pairs could be transferable to Rosa germplasms and 12 pairs were polymorphic among the 16 R. roxburghii germplasms. The results indicated that the unigenes generated from transcriptome sequencing in R. roxburghii can be used as an effective source todevelopment SSR markers. The large quantities of SSR markers will provide more reliable markers for map structure,analysis of genetic polymorphism for R. roxburghii and its closely related species.

Key words: Rosa roxburghii, SSR, transcriptome

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