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梨EST-SSR标记的开发及其在梨品种遗传多样性分析中的应用评价
引用本文:王西成,姜淑苓,上官凌飞,曹玉芬,乔玉山,章镇,房经贵.梨EST-SSR标记的开发及其在梨品种遗传多样性分析中的应用评价[J].中国农业科学,2010,43(24):5079.
作者姓名:王西成  姜淑苓  上官凌飞  曹玉芬  乔玉山  章镇  房经贵
作者单位:(南京农业大学园艺学院);
摘    要: 【目的】分析梨EST中SSR位点分布规律,开发梨EST-SSR引物,探讨EST-SSR用于梨品种遗传差异研究的可行性。【方法】从NCBI公共数据库中下载梨表达序列标签(expressed sequence tag,EST)1 293条,利用MISA软件对其进行SSR位点查找,将符合条件的序列选出,利用Primer3.0 Plus软件设计48对引物,通过非变性聚丙烯酰胺凝胶(PAGE)研究这些SSR引物的PCR扩增特点,并对部分扩增产物克隆与测序,以验证其真实性。【结果】1 293条梨的EST序列中含有SSR位点的序列为82条,SSR位点92个。二核苷酸、三核苷酸和六核苷酸重复是最主要的SSR类型,分别占48.91%、17.39%和17.39%。48对引物中有31对引物能扩增出理想的PCR 产物,其中27对引物扩增条带具有多态性。同时发现11对引物中,有83.87%的片段具有相应的SSR位点。聚类结果表明,梨被明显地区分成东方梨和西方梨两大群,分类效果明显。【结论】梨EST-SSR标记开发效率较高,是梨SSR标记开发的重要措施,对于梨品种的鉴定与遗传多样性分析具有重要应用价值。

关 键 词:  EST  SSR  遗传多样性
收稿时间:2010-05-12;

Development of EST-Derived SSR Markers for Pear and Evaluation of Their Application in Pear Genetic Diversity Analysis
WANG Xi-cheng,JIANG Shu-ling,SHANGGUAN Ling-fei,CAO Yu-fen,QIAO Yu-shan,ZHANG Zhen,FANG Jing-gui.Development of EST-Derived SSR Markers for Pear and Evaluation of Their Application in Pear Genetic Diversity Analysis[J].Scientia Agricultura Sinica,2010,43(24):5079.
Authors:WANG Xi-cheng  JIANG Shu-ling  SHANGGUAN Ling-fei  CAO Yu-fen  QIAO Yu-shan  ZHANG Zhen  FANG Jing-gui
Institution:(College of Horticulture, Nanjing Agricultural University)
Abstract:【Objective】 The object of this study was to analyze the SSR distribution in ESTs of pear and develop new EST-derived SSR markers, and application of EST-SSR markers in pear genetic analysis was also validated. 【Method】 All the 1 293 EST sequences of pear were obtained from NCBI. These sequences were screened by using MISA software to search for SSR motifs. Forty-eight pairs of primers were designed by the software Primer3.0 Plus. The PCR products of these primers were detected by PAGE and some of them were recovered for sequencing. 【Result】 The results showed that 92 SSRs were identified from the 82 pear EST sequences. The dinucleotide,trinucleotide and hexanucleotide repeats were the dominant types with the frequency of 48.91%, 17.39% and 17.39%, respectively. Among the 48 EST-SSR primers, 31 amplified distinct bands and expected products, and 27 were polymorphic. There were 83.87% of the PCR products contained the SSR markers. The dendrogram showed that 16 pear cultivars were obviously classified into 2 groups, including the Asian pear and the European pear. 【Conclusion】 Due to the high efficiency of developing SSR markers from pear ESTs, this method will be of great importance in pear genetic analysis.
Keywords:pear  EST  SSR  genetic diversity
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