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柑橘苗期SSR鉴定分析体系的优化及初步应用
引用本文:韩国辉,魏旭,向素琼,汪卫星,何波,梁国鲁.柑橘苗期SSR鉴定分析体系的优化及初步应用[J].西南农业学报,2011,24(2).
作者姓名:韩国辉  魏旭  向素琼  汪卫星  何波  梁国鲁
作者单位:南方山地园艺学教育部重点实验室,西南大学园艺园林学院,重庆400716
基金项目:重庆市重大科技专项,教育部基本科研业务费项目,西南大学研究生科技创新基金项目
摘    要:本文以柑橘幼苗微量叶片(3~5 mg)为材料,改进柑橘基因组DNA提取方法,并对3种PAGE凝胶浓度和3种银染色方法进行了筛选和优化,建立了一种经济,高效的柑橘苗期SSR分析体系.试验结果表明:该微量叶片提取方法需材少,用时短,所提取基因组DNA质量好、纯度高,完全适用于SSR-PCR扩增反应.9%的PAGE凝胶浓度最适合柑橘SSR标记的电泳检测,扩增出的条带清晰,分散良好.改进的两种快速银染方法相对于传统的染色方法染色效果较好,且更加经济.初步对两个柑橘杂交幼苗群体进行了遗传鉴定和分析,2对引物从24株沙田柚与强德勒柚杂交幼苗中鉴定出17株杂种,4对引物在沙田柚与红江橙杂交群体中扩增出14条多态性条带,占总条带数的70.0%.表明所建立的SSR优化体系可为柑橘分子辅助育种提供技术参考.

关 键 词:柑橘  苗期  SSR体系  优化

Optimization of SSR Analysis System of Citrus and Its Primary Application
HAN Guo-hui,WEI Xu,XIANG Su-qiong,WANG Wei-xing,HE Bo,LIANG Guo-lu.Optimization of SSR Analysis System of Citrus and Its Primary Application[J].Southwest China Journal of Agricultural Sciences,2011,24(2).
Authors:HAN Guo-hui  WEI Xu  XIANG Su-qiong  WANG Wei-xing  HE Bo  LIANG Guo-lu
Institution:HAN Guo-hui,WEI Xu,XIANG Su-qiong,WANG Wei-xing,HE Bo,LIANG Guo-lu(Key Laboratory of Horticulture Science for Southern Mountainous Regions,Ministry of Education/College of Horticulture and Landscape Architecture,Southwest University,Chongqing 400715,China)
Abstract:In this study,a new method to isolate DNA from microamount of citrus leaves(3-5 mg)rapidly was built.Three PAGE concentrations and three silver staining methods for citrus SSR analysis system were screened and optimized.In this way,an optimized SSR analysis technology was obtained.The results showed that the method had the characteristics of little material and shorter time,and the DNA had good quality and high purity.The 9 % PAGE was the most suitable for citrus SSR analysis and the amplified profiles were...
Keywords:Citrus  Seedling stage  SSR system  Optimization  
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