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《Journal of Fruit Science》 2009-02
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Optimization of SRAP-PCR system and its application in genetic diversity analysis of cherry

LU Juan1,ZHANG Shao-ling1,LIU Qing-zhong2,WU Jun1,ZHANG Rui-ping1,LIAOYANG Wen-ke1,CHEN Ting1(1College of Horticulture,Nanjing Agricultural University,Nanjing,Jiangsu 210095 China;2Shandong Institute of Pomology,Tai'an,Shan-dong 271000 China)  
Three different genotypes of cherry germplasm showing distant relationship were used as material for studying the effects of concentrations of Mg2+,dNTPs,Taq DNA polymerase,primers and DNA template on the SRAP-PCR reactions and optimizing the establishment of SRAP molecular marker system in cherry.The optimum system was established as follows:template DNA 75 ng,dNTPs 0.2 mmol·L-1,Mg2+ 2.5 mmol·L-1,primer 0.3 μmol·L-1,Taq polymerase 1.0 U,the total reac-tion volume was 20 μL.Amplifications were carried out on 45 samples with eight primer combinations;using this optimum system.227 steady and reliable bands were amplified,of which 192(84.6%) were polymorphic.UPGMA cluster analysis was performed by soft NTYSIS-pc 2.01,the clades formed including two groups at the dice coefficient of 0.52,namely sweet cherry(Prunus avium L.) and Chinese cherry(P.pseudocerasus L.).The genetic similarity coefficient of the 45 accessions ranged from 0.52 to 0.98.The lowest genetic similarity coefficient existed between sweet cherry and Chinese cherry;it showed that the genetic background of two groups differed from each other.The high similarity coefficient existed among dif-ferent cultivars belonging to the same groups.The cluster results revealed by SRPA showed that the relationship of cherry germplasm was related to geographic distribution and originals.
【Fund】: 中国博士后科学基金资助项目2005037740;; 江苏省博士后科学基金资助项目;; 南京农业大学青年启动基金
【CateGory Index】: S662.5
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