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):27-34.[doi:10.11937/bfyy.20181074]
白檀SRAP-PCR体系优化及引物筛选
- Title:
- Optimization and Primers Screening of SRAP-PCR System in Symplocos paniculata
- Keywords:
- Symplocos paniculata; SRAP; system optimization; primers screening
- 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075 U、dNTPs 01 mmol·L-1、Mg2+ 2 mmol·L-1、引物04 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 075 U Taq DNA polymerase,01 mmol·L-1 dNTPs,2 mmol·L-1 Mg2+,04 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.
相似文献/References:
[1]王晶,张晓明,闫国华,等.草原樱桃和对樱杂交后代的分子鉴定[J].北方园艺,2012,36(01):116.
WANG Jing,ZHANG Xiao-ming,YAN Guo-hua,et al.Molecular Identification of Hybrids between Prunus fruticosa and Prunus pseudocerasus by SRAP Method[J].Northern Horticulture,2012,36(24):116.
[2]刘小飞,廖飞雄,刘晓荣,等.鸡冠花诱变选育系和栽培选育系的SRAP分子鉴定[J].北方园艺,2013,37(07):128.
LIU Xiao-fei,LIAO Fei-xiong,LIU Xiao-rong,et al.Genetic Identification of Celosia Mutant and Select Lines Using Sequence-related Amplified Polymorphism,SRAP[J].Northern Horticulture,2013,37(24):128.
[3]郝凤,于铁峰,雷银川.番茄SRAP-PCR反应体系建立与优化[J].北方园艺,2012,36(03):107.
HAO Feng,YU Tie-feng,LEI Yin-chuan.Optimization for SRAP-PCR System of Tomato Based on Orthogonal Design[J].Northern Horticulture,2012,36(24):107.
[4]王骞春,黄夏,李光森.小干松SRAP-PCR反应体系的建立[J].北方园艺,2013,37(02):83.
WANG Qian-chun,HUANG Xia,LI Guang-sen.Establishment of the SRAP-PCR Reaction System of Pinus contorta[J].Northern Horticulture,2013,37(24):83.
[5]刘 巍,陈 罡,颜廷武,等.美国红枫SRAP反应体系优化[J].北方园艺,2015,39(02):79.[doi:10.11937/bfyy.201502023]
LIU Wei,CHEN Gang,YAN Ting-wu,et al.Optimization of SRAP Reaction System in Acer rubrum[J].Northern Horticulture,2015,39(24):79.[doi:10.11937/bfyy.201502023]
[6]毛 伟,李培英,孙宗玖.偃麦草SRAP-PCR反应体系优化[J].北方园艺,2015,39(02):91.[doi:10.11937/bfyy.201502026]
MAO Wei,LI Pei-ying,SUN Zong-jiu.Optimization of SRAP-PCR System on Elytrigia repens[J].Northern Horticulture,2015,39(24):91.[doi:10.11937/bfyy.201502026]
[7]梁婷婷,马 燕,臧德奎.梨属野生资源遗传多样性的SRAP分析[J].北方园艺,2015,39(13):1.[doi:10.11937/bfyy.201513001]
LIANG Ting-ting,MA Yan,ZANG De-kui.Genetic Diversity Analysis of Wild Germplasm Resource of Pyrus Based on SRAP Markers[J].Northern Horticulture,2015,39(24):1.[doi:10.11937/bfyy.201513001]
[8]赵英,黄国弟,付海天,等.十二份杧果种质亲缘关系的SRAP分析[J].北方园艺,2015,39(21):99.[doi:10.11937/bfyy.201521026]
ZHAO Ying,HUANG Guodi,FU Haitian,et al.SRAP Analysis of 12 Mango Germplasm Genetic Relationships[J].Northern Horticulture,2015,39(24):99.[doi:10.11937/bfyy.201521026]
[9]刘立言,宋航,安佰义,等.白檀种子发芽特性研究[J].北方园艺,2016,40(10):78.[doi:10.11937/bfyy.201610020]
LIU Liyan,SONG Hang,AN Baiyi,et al.Study on the Germination Characters of Symplocos paniculata Seeds[J].Northern Horticulture,2016,40(24):78.[doi:10.11937/bfyy.201610020]
[10]谭江平,曾秀丽.,廖明安,等.西藏光核桃SRAJ—PCR反应体系的优化和引物筛选[J].北方园艺,2011,35(02):0.[doi:10.11937/bfyy.201102049]
TAN Jiang-pingI,ZENG Xiu-li.,LIAO Ming-anI,et al.Optimization for SRAP-PCR System and Selection of Primers on Prunusmira Koehne from Tibet[J].Northern Horticulture,2011,35(24):0.[doi:10.11937/bfyy.201102049]
[11]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.[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.[doi:10.11937/bfyy.20181074]
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