基于沙棘转录组序列开发EST-SSR分子标记

[目的]本研究通过对蒙古沙棘＂向阳＂品种的转录组序列进行SSR引物开发,为沙棘亲本分析、遗传多样性和遗传育种等研究提供支持。[方法]利用已测得的转录组序列进行分析和筛选,整理具有SSR位点的序列,进行引物设计,随机挑选179条引物进行验证。[结果]得到具有SSR位点的EST序列17 383条,其中,单核苷酸、二核苷酸和三核苷酸重复基元分别占62.77%、21.82%和13.77%;二核苷酸重复基元类型以AT和AG为主,三核苷酸重复基元类型以AAG、ATC和ATA为主。9 291条序列设计出扩增EST-SSR位点的引物,随机合成的179对引物中,142对扩增成功,选取其中92个位点的扩增产物上机...

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Bibliographic Details
Published in林业科学研究 Vol. 30; no. 1; pp. 69 - 74
Main Author 李珊珊 曾艳飞 何彩云 张建国
Format Journal Article
LanguageChinese
Published 国家林业局林木培育重点实验室，中国林业科学研究院林业研究所，北京 100091%国家林业局林木培育重点实验室，中国林业科学研究院林业研究所，北京 100091 2017
南京林业大学南方现代林业协调创新中心，江苏 南京 210037
Subjects
EST-SSR
引物
沙棘
转录组
沙棘
Hippophae rhamnoides
primer
transcriptome
EST -SSR
转录组
EST-SSR
引物
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ISSN1001-1498
DOI10.13275/j.cnki.lykxyj.2017.01.010

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Summary:[目的]本研究通过对蒙古沙棘＂向阳＂品种的转录组序列进行SSR引物开发,为沙棘亲本分析、遗传多样性和遗传育种等研究提供支持。[方法]利用已测得的转录组序列进行分析和筛选,整理具有SSR位点的序列,进行引物设计,随机挑选179条引物进行验证。[结果]得到具有SSR位点的EST序列17 383条,其中,单核苷酸、二核苷酸和三核苷酸重复基元分别占62.77%、21.82%和13.77%;二核苷酸重复基元类型以AT和AG为主,三核苷酸重复基元类型以AAG、ATC和ATA为主。9 291条序列设计出扩增EST-SSR位点的引物,随机合成的179对引物中,142对扩增成功,选取其中92个位点的扩增产物上机检测,40个SSR位点（43.48%）呈现多态。对扩增稳定且峰图清晰的17个多态位点的进一步分析,得到蒙古沙棘天然群体在各位点的观测杂合度（HO）为0.083 0.875,期望杂合度（HE）为0.180 0.750。[结论]本研究开发出的EST-SSR标记,为后期进行沙棘属植物遗传多样性分析、遗传图谱构建及分子育种等研究提供了重要支持。
Bibliography:11-1221/S
Hippophae rhamnoides; transcriptome; EST-SSR; primer
[Objective]The transcriptomes of Hippophae rhamnoides subsp. mongolica cv‘Xiangyang＇were analyzed to design primers and develop EST-SSR（ Expressed Sequence Tags-Simple Sequence Repeat） markers.[Method]The primers were designedand the SSR was developed based on the Expressed Sequence. 179 pairs of primers were validated randomly. [Result]17 383 SSR-ESTs（ SSR-containing EST） were identified. Of the total EST-SSRs,the mononucleotiderepeat was the most dominant type,accounting for 62. 77%,followed by dinucleotiderepeats and trinucleotiderepeats,accounting for 21. 82% and 13. 77%,respectively. AT and AG were the most abundant dinucleotide motifs; AAG,ATC and ATA were repeated dominant motifs in trinucleotide. Based on these SSR-ESTs,9 291 pairs of EST-SSR primers were designed; 179 pairs of primers were randomly selected and compounded for PCR amplification,among which 142 loci were amplified successfully. Amplificationproductions of 92 loci were genotyp
ISSN:1001-1498
DOI:10.13275/j.cnki.lykxyj.2017.01.010