白檀SRAP-PCR体系优化及引物筛选  

Optimization and Primers Screening of SRAP-PCR System in Symplocos paniculata

作　　者：安佰义[1] 于慧颖 吴双[1] 刘晓嘉[1] 孙晓刚[1] 张艳[2] AN Baiyi;YU Huiying;WU Shuang;LILT Xiaojia;SUN Xiaogang;ZHANG Yan(College of Horticulture,Jilin Agricultural University,Changchun,Jilin 130118;Agro-biotechnology Research Institute,Jilin Academy of Agricultural Sciences,Changchun,Jilin 130124)

机构地区：[1]吉林农业大学园艺学院,吉林长春130118 [2]吉林省农业科学院农业生物技术研究所,吉林长春130124

出　　处：《北方园艺》2018年第24期15-20,共6页Northern Horticulture

基　　金：长春市科技局资助项目(17DY014);吉林省教育厅资助项目(JJKH20180665KJ);吉林省科技厅资助项目(20150204045NY)

摘　　要：以白檀新生嫩叶为试材,通过单因素试验与正交实验,建立并优化白檀SRAP-PCR反应体系,对白檀SRAP反应体系中5个因素进行优化试验,并筛选SRAP引物组合,以期为今后SRAP分子标记在白檀上的应用提供依据。结果表明:各因素对SRAP-PCR扩增结果影响差异较大,依次为模板DNA>dNTPs>Mg^(2+)>引物>Taq DNA聚合酶。最优反应体系为总体系10μL体系中,Taq DNA聚合酶0.75U、dNTPs 0.1mmol·L^(-1)、Mg^(2+)2mmol·L^(-1)、引物0.4μmol·L^(-1)以及模板DNA 80ng。利用稳定的SRAP-PCR体系,从176对引物组合中筛选出25对多态性好的引物组合。在所建立的最优体系下,不同白檀基因组和不同引物组合均能稳定扩增,表明体系稳定可靠适用于白檀SRAP分子标记研究。In order to establish and optimize SRAP-PCR reaction system in Symplocos paniculata(Thunb.)Miq.,the fresh leaves of Symplocos paniculata were used as experimental materials,the combination of single factor test and orthogonal test were used to optimize SRAP-PCR reaction system with 5factors,and SRAP primer combinations were screened.The results showed that,the influence of each factor on the result of SRAP-PCR amplification was quite different,and the descending order was DNA>dNTPs>Mg2+>primer>Taq DNA polymerase.A suitable SRAP-PCR system for Symplocos paniculata was that total 10μL reaction system containing 0.75 U Taq DNA polymerase,0.1mmol·L-1 dNTPs,2 mmol·L-1Mg2+,0.4μmol·L-1 primer and 80ng DNA.A total of 25 polymorphic SRAP primer combinations were screened from 176SRAP primer combinations.PCR products were stably amplified among plants by different primers,which indicated that the SRAP-PCR system was suitable for molecular markers research of Symplocos paniculata.

关 键 词：白檀 SRAP 体系优化 引物筛选 

分 类 号：S792.99[农业科学—林木遗传育种]