[1]郭磊,上官凌飞,房经贵*,等.葡萄EST鄄SSR 标记的开发及其应用[J].南京农业大学学报,2011,34(4):23.[doi:10.7685/j.issn.1000-2030.2011.04.005]
 GUO Lei,SHANGGUAN Ling-fei,FANG Jing-gui *,et al.Development of SSR markers from grape ESTs and its application[J].Journal of Nanjing Agricultural University,2011,34(4):23.[doi:10.7685/j.issn.1000-2030.2011.04.005]
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葡萄EST鄄SSR 标记的开发及其应用()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
34卷
期数:
2011年4期
页码:
23
栏目:
`s\乿l VE`s\乿l V="publishdate">
出版日期:
2011-08-31

文章信息/Info

Title:
Development of SSR markers from grape ESTs and its application
文章编号:
1000-2030(2011)04-0023-08
作者:
郭磊1上官凌飞1房经贵1*刘崇怀2于华平1
1. 南京农业大学园艺学院/ 江苏省果树品种改良与种苗繁育工程中心,江苏南京210095;2. 中国农业科学院郑州果树研究所,河南郑州450009
Author(s):
GUO Lei 1SHANGGUAN Ling-fei 1FANG Jing-gui 1*LIU Chong-huai 2YU Hua-ping 1
1.College of Horticulture/Fruit Grop Genetic Improvement and Seeding Propagation Engineering Center of Jiangsu Province, Nanjing Agricultural University,Nanjing 210095,China;2.Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences,Zhengzhou 450009,China
关键词:
葡萄ESTSSR遗传多样性
Keywords:
grapeESTSSRgenetic diversity
分类号:
S663.1
DOI:
10.7685/j.issn.1000-2030.2011.04.005
文献标志码:
A
摘要:
NCBI 公共数据库中随机下载葡萄表达序列标签 (expressed sequence tag,EST)8 925 , 利用 Phrap 软件对这些序列进行拼接后形成 5 642 条序列 , 利用 MISA 软件共发掘出含有 SSR 位点的序列 336 , 共含有 367 SSR 位点 , 候选 SSR 位点出现的频率为 6.50% 。二核苷酸、三核苷酸、四核苷酸、五核苷酸和六核苷酸重复单元的 SSR 数目分别为 79(21.53%) 83(22.62%) 29(7.9%) 60(16.35%) 116(31.61%) 。利用 Primer 3.0 Plus 软件随机设计 50 对引物进行 PCR 扩增 , 6% 非变性聚丙烯酰胺凝胶电泳分析这些 SSR 引物的 PCR 扩增产物多态性。 50 对引物中有 36 对引物能在 20 个葡萄品种中扩增出理想的 PCR 产物 , 占总引物的 72% 。其中 ,26 对引物扩增条带具有多态性。利用 16 对经过验证的 EST-SSR 引物对 20 个葡萄品种亲缘关系的分析获得了理想的研究结果。
Abstract:

A total of 8 925 grapevine EST(expressed sequence tag)sequences were randomly downloaded from NCBI,and were further assembled into 5 642 uniform sequences using Phrap software.These sequences were screened by using MISA software to search for SSR motifs and 367 SSR loci were identified from the 336 grape EST sequences.The frequency of EST-SSR identified was 6.50%.All the 367 SSR loci included 79(21.53%)dinucleotides,83(22.62%)trinucleotides,29(7.9%)tetranucleotides,60(16 . 35%) pentanucleotides,and 116(31.61%)hexanucleotides.Fifty pairs of primers were designed against the some EST sequences by the software Primer 3.0 Plus,and the PCR products of these primers were detected by PAGE and some of them were collected for sequencing.Among the 50 pairs of EST-SSR primers,36 ones could amplify distinct PCR bands that were the anticipated products.Twenty-six pairs of the EST-SSR primers could amplify polymorphic bands.With sequencing the PCR products,sixteen pairs of primers were verified to be able to amplify real SSR sequences,and the dendrogram of 20 grapevine cultivars generated with the polymorphic marker information from these 16 pairs of EST-SSR primers showed that these cultivars were relatively classified well.


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备注/Memo

备注/Memo:
收稿日期:2010-11-10
基金项目:江苏省科技支撑项目(BE2010326);第36批留学回国科研启动基金项目
作者简介:郭磊,硕士研究生。*通讯作者:房经贵,教授,博导,主要从事果树基因组学与分子生物学研究,E-mail:fanggg@njau.edu.cn。
更新日期/Last Update: 2011-09-12