草莓属植物SCoT分析体系的建立及优化Establishment and optimization of SCoT-PCR system in Fragaria
秦国新;何桥;梁国鲁;雷家军;郭启高;
摘要(Abstract):
为利用SCoT技术开展草莓属(Fragaria)植物种质资源遗传多样性及分子系统学研究,以草莓属植物11个野生种和1个栽培品种为试材,采用L25(56)正交试验设计,建立了草莓属植物的最优SCoT-PCR反应体系,即20μL反应体系中,含Mg2+2.0 mmol·L-1、dNTPs 0.3 mmol·L-1、Taq酶0.75 U、引物0.75μmol·L-1、模板DNA 40 ng,并对退火温度进行了优化,对电泳检测方法及草莓属植物DNA提取方法进行了改进。共筛选出22个适于进行草莓属植物SCoT分析的引物。经验证,该反应体系稳定性好、可重复性强、多态性高、扩增效果好,适于草莓属植物的SCoT分析。
关键词(KeyWords): 草莓属;SCoT;体系优化
基金项目(Foundation): 国家自然科学基金(30971976)
作者(Author): 秦国新;何桥;梁国鲁;雷家军;郭启高;
Email:
DOI: 10.13925/j.cnki.gsxb.2012.03.009
参考文献(References):
- [1]LEI Jia-jun,DAI Han-ping,TAN Chang-hua,DENG Ming-qin,ZHAO Mi-zhen,QIAN Ya-ming.Studies on the taxonomy of thestrawberry(Fragaria)species distributed in China[J].Acta Horticul-turae Sinica,2006,33(1):1-5.雷家军,代汉萍,谭昌华,邓明琴,赵密珍,钱亚明.中国草莓属(Fragaria)植物的分类研究[J].园艺学报,2006,33(1):1-5.
- [2]SHI Cui-ping,GE Hui-bo,ZHANG Cheng-he,GUO Zhen-huai.Cytological studies on unreduced gamete formation of strawberries(Fragaria)[J].Scientia Agricultura Sinica,2002,35(10):1260-1263.时翠平,葛会波,张成合,郭振怀.草莓未减数配子形成的细胞学研究[J].中国农业科学,2002,35(10):1260-1263.
- [3]DENG Ming-qin,LEI Jia-jun.China fruit records-Volume straw-berry[M].Beijing:China Forest Press,2005,4:15-23.邓明琴,雷家军.中国果树志.草莓卷[M].北京:中国林业出版社,2005,4:15-23.
- [4]WANG Zhi-gang,ZHANG Zhi-hong.Application of molecularmarkers on genetic research of strawberry[J].Liaoning AgriculturalSciences,2005(3):36-38.王志刚,张志宏.分子标记在草莓遗传研究中的应用[J].辽宁农业科学,2005(3):36-38.
- [5]CHEN Qiong-e,ZHONG Feng-lin,PAN Dong-ming,LI Kai-tuo.Progress of molecular mark on genetic research of strawberry[J].Sub-tropical Agriculture Research,2011,7(1):64-67.陈琼娥,钟凤林,潘东明,李开拓.草莓分子标记技术研究进展[J].亚热带农业研究,2011,7(1):64-67.
- [6]COLLARD B C Y,MACKILL D J.Start Codon Targeted(SCoT)Polymorphism:A simple,novel DNA Marker technique for generat-ing gene-Targeted markers in plants[J].Plant Mol Biol Rep,2009,27:86-93.
- [7]XIONG Fa-qian,TANG Rong-hua,CHEN Zhong-liang,PAN Ling-hua,ZHUANG Wei-jian.SCoT:A novel gene targeted marker tech-nique based on the translation start codon[J].Molecular Plant Breed-ing,2009,7(3):635-638.熊发前,唐荣华,陈忠良,潘玲华,庄伟建.目标起始密码子多态性(SCoT):一种基于翻译起始位点的目的基因标记新技术[J].分子植物育种,2009,7(3):635-638.
- [8]CHEN Hu,HE Xin-hua,LUO Cong,GAO Mei-ping,ZHU Jian-hua.The Optimization of SCoT-PCR System of Longan(Dimocarpus lon-gan)[J].Genomics and Applied Biology,2009,28(5):970-974.陈虎,何新华,罗聪,高美萍,朱建华.龙眼SCoT-PCR反应体系的优化[J].基因组学与应用生物学,2009,28(5):970-974.
- [9]CHEN Hu,HE Xin-hua,LUO Cong,ZHU Jian-hua,LI Feng.Analy-sis on the Genetic Diversity of 24 Longan(Dimocarpus longan)Acces-sions by SCoT Markers[J].Acta Horticulturae Sinica,2010,37(10):1651-1654.陈虎,何新华,罗聪,朱建华,李峰.龙眼24个品种的SCoT遗传多样性分析[J].园艺学报,2010,37(10):1651-1654.
- [10]HAN Guo-hui,WANG Wei-xing,XIANG Su-qiong,BIAN Yu,GUOQi-gao,HE Qiao,LI Xiao-lin,LIANG Guo-lu.Establishment andoptimization of SCoT system in polyploidy loquats[J].Journal of FruitScience,2011,28(3):433-437.韩国辉,汪卫星,向素琼,边禹,郭启高,何桥,李晓林,梁国鲁.多倍体枇杷SCoT分析体系的建立与优化[J].果树学报,2011,28(3):433-437.
- [11]HAN Guo-hui,XIANG Su-qiong,WANG Wei-xing,JIA Zhi-gang,HONG Qi-bin,LIANG Guo-lu.Establishment and application ofSCoT molecular marker system for Citrus[J].Acta Horticulturae Sini-ca,2011,38(7):1243-1250.韩国辉,向素琼,汪卫星,贾志刚,洪棋斌,梁国鲁.柑橘SCoT分子标记技术体系的建立及其在遗传分析中的应用[J].园艺学报,2011,38(7):1243-1250.
- [12]ZHANG Jun-yu,GUO Da-long,GONG Ying,LIU Chong-huai,LIMeng,ZHANG Guo-hai.Optimization of start codon targeted poly-morphism PCR(SCoT-PCR)system in Vitis vinifera[J].Journal ofFruit Science,2011,28(2):209-214.张君玉,郭大龙,龚莹,刘崇怀,李猛,张国海.葡萄目标起始密码子多态性反应体系的优化[J].果树学报,2011,28(2):209-214.