[1]安佰义,于慧颖,吴双,等.白檀SRAP-PCR体系优化及引物筛选[J].北方园艺,2018,42(24):15-20.[doi:10.11937/bfyy.20181074]
　AN Baiyi,YU Huiying,WU Shuang,et al.Optimization and Primers Screening of SRAP-PCR System in Symplocos paniculata[J].Northern Horticulture,2018,42(24):15-20.[doi:10.11937/bfyy.20181074]
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白檀SRAP-PCR体系优化及引物筛选

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第42卷 期数: 2018年24 页码: 15-20 栏目: 出版日期: 2018-12-25
Title:
Optimization and Primers Screening of SRAP-PCR System in Symplocos paniculata
作者:
安佰义1; 于慧颖1; 吴双1; 刘晓嘉1; 孙晓刚1; 张艳2
（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）
Author(s):
AN Baiyi1; YU Huiying1; WU Shuang1; LIU Xiaojia1; SUN Xiaogang1; ZHANG Yan2
(1.College of Horticulture,Jilin Agricultural University,Changchun,Jilin 130118;2.Agro-biotechnology Research Institute,Jilin Academy of Agricultural Sciences,Changchun,Jilin 130124)
关键词:
白檀; SRAP; 体系优化; 引物筛选
Keywords:
Symplocos paniculata; SRAP; system optimization; primers screening
DOI:
10.11937/bfyy.20181074
文献标志码:
A
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＞Mg2+＞引物＞Taq DNA聚合酶。最优反应体系为总体系10 μL体系中，Taq DNA聚合酶075 U、dNTPs 01 mmol·L-1、Mg2+ 2 mmol·L-1、引物04 μmol·L-1以及模板DNA 80 ng。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。
Abstract:
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 5 factors,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 10 μL reaction system containing 075 U Taq DNA polymerase,01 mmol·L-1 dNTPs,2 mmol·L-1 Mg2+,04 μmol·L-1 primer and 80 ng DNA.A total of 25 polymorphic SRAP primer combinations were screened from 176 SRAP 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.

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备注/Memo

第一作者简介：安佰义（1978-），男，山东日照人，博士，副教授，研究方向为园林植物种质创新及栽培生理与景观生态及功能基因挖掘。E-mail：swabyswaby@163.com.责任作者：孙晓刚（1969-），男，吉林长春人，硕士，教授，研究方向为园林植物应用与景观评价。E-mail:120082055@qq.com.基金项目： 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)。收稿日期：2018-05-23

更新日期/Last Update: 2019-01-22