一个纯显性RAPD标记的获得及其在龙眼杂种真实性鉴定中的应用Application of a homozygosis dominant RAPD marker in identifying longan hybrids
卢博彬;黄天林;潘丽佳;傅嘉欣;王英;刘朝吉;胡桂兵;刘成明;
摘要(Abstract):
【目的】在龙眼上,营养系杂交和杂种真实性早期鉴定的工作都还开展甚少,为了研发出既操作简便又稳定可靠的早期鉴定技术,【方法】以人工杂交获得的‘凤梨朵’ב大乌圆’的大规模F1代杂交群体(简称FD群体)中的230个单株为试材,先通过40个RAPD引物对双亲进行扩增筛选获得父本特异性的显性标记,再以父本的小规模自交群体(9株)为材料,明确这些标记在自交群体中的分离情况,最终鉴别出纯显性标记并用于对FD群体的杂种真实性鉴定。【结果】结果表明,对双亲进行扩增的RAPD引物中有9个能扩增出父本特异性的显性标记,用这9个引物检测父本自交群体的分离情况,最终鉴别出纯显性标记1个,用其对FD群体中的230个单株进行杂种真实性鉴定,发现有223个单株为真杂种,真杂种比例高达96.96%。【结论】既获得了早期鉴定杂种真实性的简便方法,又证明了创建的龙眼杂种群体非常可靠,值得在今后的育种工作中借鉴和使用。
关键词(KeyWords): 龙眼;营养系杂交育种;杂种鉴定
基金项目(Foundation): 广东省科技计划项目(2012A020200005,2009B020202001);; 广东省现代农业产业技术体系(LNSG-2011-5,LNSG-2010-5);; 广东省国际合作项目(2009B050300004);; 国家自然科学基金(30771498,30270922);; 中国博士后基金(20090460780,20080440763)
作者(Author): 卢博彬;黄天林;潘丽佳;傅嘉欣;王英;刘朝吉;胡桂兵;刘成明;
Email:
DOI: 10.13925/j.cnki.gsxb.2013.03.010
参考文献(References):
- [1]LUO Jun,ZHOU Can-fang,WAN Zhong,HE Shu-qun,CHENHou-bin.Analysis on the development of Guangdong longan indus-try in 2010[J].Guangdong Agricultural Sciences,2011(4):16-18.罗军,周灿芳,万忠,何淑群,陈厚彬.2010年广东龙眼产业发展现状分析[J].广东农业科学,2011(4):16-18.
- [2]GUO Yin-shan,ZHAO Yu-hui,LIU Chao-ji,REN Peng-rong,HUANG Tian-lin,FU Jia-xin,LU Bo-bin,LIU Cheng-ming.Construction of a molecular genetic map for longan based on RAPD,ISSR,SRAP and AFLP markers[J].Acta Horticulturae Sinica,2009,36(5):655-662.郭印山,赵玉辉,刘朝吉,任鹏荣,黄天林,傅嘉欣,卢博彬,刘成明.利用多种分子标记构建龙眼高密度分子遗传图谱[J].园艺学报,2009,36(5):655-662.
- [3]ZHAO Yu-hui,GUO Yin-shan,HU You-li,ZHANG Bin,LIURui,OUYANG Ruo,FU Jia-xin,LIU Cheng-ming.Construction ofa genetic linkage map of litchi with RAPD,SRAP and AFLP[J].ActaHorticulturae Sinica,2010,37(5):697-704.赵玉辉,郭印山,胡又厘,张斌,刘睿,欧阳若,傅嘉欣,刘成明.应用RAPD、SRAP及AFLP标记构建荔枝高密度复合遗传图谱[J].园艺学报,2010,37(5):697-704.
- [4]ZHAO Yu-hui,GUO Yin-shan,FU Jia-xin,ZHOU Jia,HUANGSui-sheng,LIU Cheng-ming.Establishment and optimization ofSRAP reaction system in longan[J].Chinese Agricultural ScienceBulletin,2009,25(18):409-412.赵玉辉,郭印山,傅嘉欣,周佳,黄穗生,刘成明.龙眼SRAP反应体系的建立和优化[J].中国农学通报,2009,25(18):409-412.
- [5]ZHAO Yu-hui,HU You-li,GUO Yin-shan,ZHOU Jia,FU Jia-xin,LIU Cheng-ming,ZHU Jiang,ZHANG Ming-jun.Inter-generic hybrids obtained from cross between litchi and longan culti-vars and their molecular identification[J].Journal of Fruit Science,2008,25(6):950-952.赵玉辉,胡又厘,郭印山,周佳,傅嘉欣,刘成明,朱江,张明君.荔枝、龙眼属间远缘杂种的获得及分子鉴定[J].果树学报,2008,25(6):950-952.
- [6]LIU Cheng-ming,MEI Man-tong.Researches on DNA isolation andoptimal parameters of RAPD in some Sapindaceae fruit trees[C]//Chinese Society for Horticultural Science.5th Youth Symposium onChinese Horticultural Science.Beijing:CSHS,2002:297-302.刘成明,梅曼彤.无患子科果树植物DNA的高效提取及RAPD分析体系的优化[C]//中国园艺学会.中国园艺学会第五届青年学术讨论会论文集.北京:中国园艺学会,2002:297-302.
- [7]LIU Cheng-ming,MEI Man-tong.Identification of stenospermo-carpy mutants of litchi by RAPD analysis[J].Acta Horticulturae Sini-ca,2002,29(1):57-59.刘成明,梅曼彤.利用RAPD分析鉴别荔枝的焦核突变体[J].园艺学报,2002,29(1):57-59.
- [8]LIU Chao-ji,FU Jia-xin,FAN Ming-he,GUO Yin-shan,ZENGLiang-shun,HUANG Tian-lin,SU Xu,SU Zhang-cheng,LIUCheng-ming,HUANG Sui-sheng.Preliminary study on longan arti-ficial crossing and several traits of the hybrid populations[J].ActaHorticulturae,2010,863:161-168.
- [9]MURRAY M G,THOMPSON W F.Rapid isolation of high molecularweight plant DNA[J].Nucleic Acids Research,1980,8:4321-4325.
- [10]STEPHEN L D,JONATHAN W,JAMES B H.A plant DNAminipreparation:Version II[J].Plant Molecular Biology Reporter,1983,1(4):19-21.
- [11]WILLIAMS J G K,KUBELIK A R,LIVAK KJ,RAFALLSKI JA,Tingey SV.DNA polymorphisms amplified by arbitrary primers areuseful as genetic markers[J].Nucleic Acids Research,1990,18(22):6531-6535.
- [12]TIAN Lei,GUAN Rong-xia,LIU Zhang-xiong,CHANG Ru-zhen,QIU Li-juan.Verity identification of soybean hybrids using SSRmarkers[J].Journal of Plant Genetic Resources,2008,9(4):443-447.田蕾,关荣霞,刘章雄,常汝镇,邱丽娟.用SSR标记鉴定大豆杂交组合F1的方法研究[J].植物遗传资源学报,2008,9(4):443-447.
- [13]GUO Yin-shan,LIU Chao-ji,ZHAO Yu-hui,XIE Jiang-hui,REN Peng-rong,HUANG Tian-lin,FU Jia-xin,LU Bo-bin,LIUCheng-ming.Construction of a molecular genetic linkage map forlongan based on RAPD,ISSR,SRAP and AFLP markers[J].ActaHorticulturae,2010,863:141-147.
- [14]GUO Yin-shan,ZHAO Yu-hui,LIU Cheng-ming.QTLs analysis ofseveral traits in longan,Biotechnol[J].&Biotechnol Eq,2011,25(1):2203-2209.
- [15]GUO Yin-shan,ZHAO Yu-hui,FU Jia-xin,HUANG Sui-sheng,WANG Ying,LU Bo-bin,PAN Li-jia,LIU Cheng-ming.Identifi-cation of stable QTLs related to trunk girth in longan[J].ScientiaHorticulturae,2012(134):248-252.