新疆主栽骨干棉花品种指纹图谱构建及纯度鉴定
|
摘要
【目的】构建新疆主栽骨干棉花品种的指纹图谱,分析棉花品种纯度,为棉花品种提供分子鉴定依据。【方法】以典型代表性的新疆北疆早熟棉花主推骨干棉花品种为材料,利用SSR分子标记技术从1000多对SSR引物中筛选出稳定性好、多态性高的25对核心引物构建指纹图谱,从核心引物中筛选10对引物作为标记,分析4个棉花品种的纯度。【结果】检测到等位基因数70个,每个标记检测到的等位基因数介于2～7个,平均2.8个;引物多态信息量(PIC)值介于0.1690～0.8729,平均值为0.6883;利用4个特征引物将4个棉花品种一次性完全区分开,构建供试品种的指纹图谱;利用10对引物检测4个品种的纯度,新陆早6号纯度变化范围85%～100%,系62纯度变化范围95%～100%。【结论】构建了4个棉花品种的指纹图谱及鉴定纯度,系62号纯度最高,为99.5%,新陆早6号纯度最低,为92.0%。
【Objective】 This project aims to construct fingerprinting map and analyze the purity of four main cotton cultivars in Xinjiang in the hope of providing a basis for molecular identification of cotton varieties.【Method】Typically representative early-mature cotton varieties in northern Xinjiang were used as materials, 25 pairs of primers with high polymorphism, good stability selected from 1,000 pairs of SSR primers as the main extension backbone varieties to construct fingerprinting map, and 10 primers were selected to analyze the purity of 4 varieties of cotton.【Result】The number of alleles detected was 70 and the number of alleles detected by each marker ranged from 2 to 7, with an average of 2.8; The polymorphism information content ranged from 0.169,0 to 0.872,9, with an average of 0.688,3; 4 specific primers could completely differentiate the 4 cotton varieties from each other and their construct fingerprinting map. 10 pairs of primers were used to detect the purity of 4 varieties, and the results showed that the variety with highest purity was Xi 62, with the purity of 99.5% and the variety with lowest purity was Xinluzao 6, whose purity was 92.0%.【Conclusion】The fingerprint and the identification purity of the four cotton varieties were constructed by the SSR molecular marker technique, and the theoretical basis for the construction and the purity identification of the fingerprint of the subsequent main varieties was established.
【Objective】 This project aims to construct fingerprinting map and analyze the purity of four main cotton cultivars in Xinjiang in the hope of providing a basis for molecular identification of cotton varieties.【Method】Typically representative early-mature cotton varieties in northern Xinjiang were used as materials, 25 pairs of primers with high polymorphism, good stability selected from 1,000 pairs of SSR primers as the main extension backbone varieties to construct fingerprinting map, and 10 primers were selected to analyze the purity of 4 varieties of cotton.【Result】The number of alleles detected was 70 and the number of alleles detected by each marker ranged from 2 to 7, with an average of 2.8; The polymorphism information content ranged from 0.169,0 to 0.872,9, with an average of 0.688,3; 4 specific primers could completely differentiate the 4 cotton varieties from each other and their construct fingerprinting map. 10 pairs of primers were used to detect the purity of 4 varieties, and the results showed that the variety with highest purity was Xi 62, with the purity of 99.5% and the variety with lowest purity was Xinluzao 6, whose purity was 92.0%.【Conclusion】The fingerprint and the identification purity of the four cotton varieties were constructed by the SSR molecular marker technique, and the theoretical basis for the construction and the purity identification of the fingerprint of the subsequent main varieties was established.
引文
[1]张玉翠.我国棉花品种SSR指纹数据库的初步构建[D].北京:中国农业科学院 ,2012 :1-8.ZHANG Yu-cui.(2012).Construction a Primary SSR Fingerprint Database for Cotton Varieties [D].Master Dissertation.Chinese Academy of Agricultural Sciences,Beijing:1-8.(in Chinese)
[2] 张玉翠,杨伟华,匡猛,等.32个棉花主栽品种 DNA 指纹图谱构建及遗传多样性分析[J].棉花学报,2012,24(2):120- 126.ZHANG Yu-cui,YANG Wei-hua,KUANG Meng,et al.(2012).Construction of DNA fingerprinting and analysis of genetic diversity with SSR markers for 32 leading cotton varieties [J].Cotton Science,24(2):120-126.(in Chinese)
[3] Zhang,H.,Wang,H.,Guo,S.,Ren,Y.,Gong,G.,& Weng,Y.,et al.(2012).Identification and validation of a core set of microsatellite markers for genetic diversity analysis in watermelon,Citrullus Lanatus Thunb.Matsum.& Nakai.Euphytica,186(2):329-342.
[4] Liu,D.,Guo,X.,Lin,Z.,Nie,Y.,& Zhang,X.(2006).Genetic diversity of asian cotton ( gossypium arboreum l.) in china evaluated by microsatellite analysis.Genetic Resources & Crop Evolution,53(6):1,145-1,152.
[5] Iqbal,M.J.,Aziz,N.,Saeed,N.A.,Zafar,Y.,& Malik,K.A.(1997).Genetic diversity evaluation of some elite cotton varieties by rapd analysis.Theoretical and Applied Genetics,94(1):139-144.
[6] 冯艳芳.基于SSR分子标记棉花品种DNA指纹图谱构建的研究[D].乌鲁木齐:新疆农业大学,2015.FENG Yan-fang.(2015).Construction of DNA fingerprint of cotton based on SSR molecular markers [D].Master Dissertation.Xinjiang Agricultural University,Urumqi.(in Chinese)
[7]贺道华,邢宏宜,李婷婷,等.92份棉花资源遗传多样性的SSR分析[J].西北植物学报,2010,30(8):1 557-1 564.HE Dao-hua,XING Hong-yi,LI Ting-ting,et al.(2010).Genetic Diversity of 92 Cotton Accessions Evaluated with SSR markers [J].Acta Botanica Boreali-Occidentalia Sinica,30(8):1,557-1,564.(in Chinese)
[8]田琴,孔宪辉,余渝,等.早熟陆地棉亲本遗传距离与杂种优势的相关性分析[J].新疆农业科学,2011,48(5):792-798.TIAN Qin,KONG Xian-hui,YU Yu,et al.(2011).Correlation Analysis of Genetic Distance and Heterosis of Early Maturing Upland Cotton Parent Line [J].Xinjiang Agricultural Sciences,48(5):792-798.(in Chinese)
[9]李育强,陈浩东,洪亚辉,等.湘杂棉SSR指纹图谱的构建及应用[J].棉花学报,2009,21(3):175-178.LI Yu-qiang,CHEN Hao-dong,HONG Ya-hui,et al.(2009).Construction and application of SSR fingerprints of Hunan hybrid cotton varieties [J].Cotton Science,21(3):175-178.(in Chinese)
[10]吴大鹏,房嫌嫌,马梦楠,等.四个国家海岛棉品种资源的亲缘关系和遗传多态性研究[J].棉花学报,2010,22(2):104-109.WU Da-peng,FANG Xian-xian,MA Meng-nan,et al.(2010).Genetic relationship and diversity of the Germplasms in Gossypium barbadense L.from four different contries using SSR markers [J].Cotton Science,22(2):104-109.(in Chinese)
[11]薛艳,李保成,张新宇,等.新疆早熟棉品种SSR分子标记体系的建立[J].石河子大学学报(自然科学版),2011,29(1):1-5.XUE Yan,LI Bao-cheng,ZHANG Xin-yu,et al.(2011).Optimization of amplification reaction system and construction of DNA fingerprinting of local upland cotton variety in Xinjiang [J].Journal of Shihezi University (Nat Sci Ed.),29(1):1-5.(in Chinese)
[12]刘国栋,王芙蓉,宫永超,等.棉花品种遗传纯度的SSR分子标记鉴定技术研究[J].棉花学报,2013,25(5):382-387.LIU Guo-dong,WANG Fu-rong,GONG Yong-chao,et al.(2013).Analysis on Gene Purity and Specificity of Cotton Varieties by SSR Marker [J].Cotton Science,25(5):382-387.(in Chinese)
[13]聂新辉,尤春源,李晓方,等.新陆早棉花品种DNA指纹图谱的构建及遗传多样性分析[J].作物学报,2014,40(12):2 104-2 117.NIE Xin-hui,YOU Chun-yuan,LI Xiao-fang,et al.(2014).Construction of DNA Fingerprinting and Analysis of Genetic Diversity for Xinluzao Cotton Varieties [J].Acta Agronomica Sinica,40(12):2,104-2,117.(in Chinese)
[14]尤春源,聂新辉,张胜,等.新疆彩色棉23个品种指纹图谱的构建及遗传多样性分析[J].棉花学报,2014,26(2):161-170.YOU Chun-yuan,NIE Xin-hui,ZHANG Sheng,et al.(2014).Construction of DNA Fingerprinting and Analysis of Genetic Diversity with SSR Markers for 23 color Cotton Cultivars in Xinjiang [J].Cotton Science,26(2):161-170.(in Chinese)
[15]李根英,Dreisigacker S,Warburton M L,等.小麦指纹图谱数据库的建立及SSR 分子标记试剂盒的研发[J].作物学报,2006,32(12):1 771-1 778.LI Gen-ying,Dreisigacker S,Warburton M L,et al.(2006).Development of a fingerprinting database and assembling an SSR reference kit for genetic diversity analysis of wheat [J].Acta Agrpmp,oca Sinica,32(12):1,771-1,778.(in Chinese)
[16] 程保山,万志兵,洪德林.35 个粳稻品种 SSR 指纹图谱的构建及遗传相似性分析[J].南京农业大学学报,2007,30(3):1-8.CHENG Bao-shan,WAN Zhi-bing,HONG De-lin,et al.(2007).Establishment of SSR fingerprint map and analysis of genetic similarity among 35 varieties in japonica rice [J].Journal of Nanjing Agricultural University,30(3):1-8.(in Chinese)
[17] 田清震,盖钧镒,喻德跃,等.我国野生大豆与栽培大豆A FLP指纹图谱研究[J].中国农业科学,2001,34(5):465 - 468TIAN Qing-zhen,GAI Jun-yi,YU De-yue,et al.(2001).AFL P fingerprint analysis of G.soja and G.max in China [J].Scientia Agricultura Sinica,34(5):465- 468.(in Chinese)
[18] 王凤格,杨扬,易红梅,等.中国玉米审定品种标准SSR指纹库的构建[J].中国农业科学,2017,50(1):1-14.WANG Feng-ge,YANG Yang,YI Hong-mei,et al.(2017).Construction of an SSR-Based Standard Fingerprint Database for Corn Variety Authorized in China [J].Scientia Agricultura Sinica,50(1):1-14.(in Chinese)
[19] 徐军,刘艳华,任民,等.普通烟草种质资源的 SSR 标记与指纹图谱分析[J].中国烟草科学,2011,32(2):62-65.XU Jun,LIU Yan-hua,REN Min,et al.(2011).SSR fingerprint map analysis of tobacco germplasms [J].Chinese Gobacco Science,32(2):62- 65.(in Chinese)
[20] 张小娟,何团结,陆徐忠,等.陆地棉SSR核心引物筛选及95份骨干种质的遗传多样性分析[J].棉花学报,2011,23(6):529-536.ZHANG Xiao-juan,HE Tuan-jie,LU Xu-zhong,et al.(2011).Screening of the SSR Core Primers of Upland Cotton and Analysis of Genetic Diversity for 95 Core Germplasms with SSR Markers [J].Cotton Science,23(6):529-536.(in Chinese)
[21] 朱美霞,李英芝,王建书,等.利用SSR方法鉴定棉花品种纯度[J].安徽农业科学,2005,33(11):2 010-2 016.ZHU Mei-xia,LI Ying-zhi,WANG Jian-shu,et al.(2005).Purity identification of cotton varieties with SSR technique [J].Journal of Anhui Agicultural Sciences,33(11):2,010-2,016.(in Chinese)
[2] 张玉翠,杨伟华,匡猛,等.32个棉花主栽品种 DNA 指纹图谱构建及遗传多样性分析[J].棉花学报,2012,24(2):120- 126.ZHANG Yu-cui,YANG Wei-hua,KUANG Meng,et al.(2012).Construction of DNA fingerprinting and analysis of genetic diversity with SSR markers for 32 leading cotton varieties [J].Cotton Science,24(2):120-126.(in Chinese)
[3] Zhang,H.,Wang,H.,Guo,S.,Ren,Y.,Gong,G.,& Weng,Y.,et al.(2012).Identification and validation of a core set of microsatellite markers for genetic diversity analysis in watermelon,Citrullus Lanatus Thunb.Matsum.& Nakai.Euphytica,186(2):329-342.
[4] Liu,D.,Guo,X.,Lin,Z.,Nie,Y.,& Zhang,X.(2006).Genetic diversity of asian cotton ( gossypium arboreum l.) in china evaluated by microsatellite analysis.Genetic Resources & Crop Evolution,53(6):1,145-1,152.
[5] Iqbal,M.J.,Aziz,N.,Saeed,N.A.,Zafar,Y.,& Malik,K.A.(1997).Genetic diversity evaluation of some elite cotton varieties by rapd analysis.Theoretical and Applied Genetics,94(1):139-144.
[6] 冯艳芳.基于SSR分子标记棉花品种DNA指纹图谱构建的研究[D].乌鲁木齐:新疆农业大学,2015.FENG Yan-fang.(2015).Construction of DNA fingerprint of cotton based on SSR molecular markers [D].Master Dissertation.Xinjiang Agricultural University,Urumqi.(in Chinese)
[7]贺道华,邢宏宜,李婷婷,等.92份棉花资源遗传多样性的SSR分析[J].西北植物学报,2010,30(8):1 557-1 564.HE Dao-hua,XING Hong-yi,LI Ting-ting,et al.(2010).Genetic Diversity of 92 Cotton Accessions Evaluated with SSR markers [J].Acta Botanica Boreali-Occidentalia Sinica,30(8):1,557-1,564.(in Chinese)
[8]田琴,孔宪辉,余渝,等.早熟陆地棉亲本遗传距离与杂种优势的相关性分析[J].新疆农业科学,2011,48(5):792-798.TIAN Qin,KONG Xian-hui,YU Yu,et al.(2011).Correlation Analysis of Genetic Distance and Heterosis of Early Maturing Upland Cotton Parent Line [J].Xinjiang Agricultural Sciences,48(5):792-798.(in Chinese)
[9]李育强,陈浩东,洪亚辉,等.湘杂棉SSR指纹图谱的构建及应用[J].棉花学报,2009,21(3):175-178.LI Yu-qiang,CHEN Hao-dong,HONG Ya-hui,et al.(2009).Construction and application of SSR fingerprints of Hunan hybrid cotton varieties [J].Cotton Science,21(3):175-178.(in Chinese)
[10]吴大鹏,房嫌嫌,马梦楠,等.四个国家海岛棉品种资源的亲缘关系和遗传多态性研究[J].棉花学报,2010,22(2):104-109.WU Da-peng,FANG Xian-xian,MA Meng-nan,et al.(2010).Genetic relationship and diversity of the Germplasms in Gossypium barbadense L.from four different contries using SSR markers [J].Cotton Science,22(2):104-109.(in Chinese)
[11]薛艳,李保成,张新宇,等.新疆早熟棉品种SSR分子标记体系的建立[J].石河子大学学报(自然科学版),2011,29(1):1-5.XUE Yan,LI Bao-cheng,ZHANG Xin-yu,et al.(2011).Optimization of amplification reaction system and construction of DNA fingerprinting of local upland cotton variety in Xinjiang [J].Journal of Shihezi University (Nat Sci Ed.),29(1):1-5.(in Chinese)
[12]刘国栋,王芙蓉,宫永超,等.棉花品种遗传纯度的SSR分子标记鉴定技术研究[J].棉花学报,2013,25(5):382-387.LIU Guo-dong,WANG Fu-rong,GONG Yong-chao,et al.(2013).Analysis on Gene Purity and Specificity of Cotton Varieties by SSR Marker [J].Cotton Science,25(5):382-387.(in Chinese)
[13]聂新辉,尤春源,李晓方,等.新陆早棉花品种DNA指纹图谱的构建及遗传多样性分析[J].作物学报,2014,40(12):2 104-2 117.NIE Xin-hui,YOU Chun-yuan,LI Xiao-fang,et al.(2014).Construction of DNA Fingerprinting and Analysis of Genetic Diversity for Xinluzao Cotton Varieties [J].Acta Agronomica Sinica,40(12):2,104-2,117.(in Chinese)
[14]尤春源,聂新辉,张胜,等.新疆彩色棉23个品种指纹图谱的构建及遗传多样性分析[J].棉花学报,2014,26(2):161-170.YOU Chun-yuan,NIE Xin-hui,ZHANG Sheng,et al.(2014).Construction of DNA Fingerprinting and Analysis of Genetic Diversity with SSR Markers for 23 color Cotton Cultivars in Xinjiang [J].Cotton Science,26(2):161-170.(in Chinese)
[15]李根英,Dreisigacker S,Warburton M L,等.小麦指纹图谱数据库的建立及SSR 分子标记试剂盒的研发[J].作物学报,2006,32(12):1 771-1 778.LI Gen-ying,Dreisigacker S,Warburton M L,et al.(2006).Development of a fingerprinting database and assembling an SSR reference kit for genetic diversity analysis of wheat [J].Acta Agrpmp,oca Sinica,32(12):1,771-1,778.(in Chinese)
[16] 程保山,万志兵,洪德林.35 个粳稻品种 SSR 指纹图谱的构建及遗传相似性分析[J].南京农业大学学报,2007,30(3):1-8.CHENG Bao-shan,WAN Zhi-bing,HONG De-lin,et al.(2007).Establishment of SSR fingerprint map and analysis of genetic similarity among 35 varieties in japonica rice [J].Journal of Nanjing Agricultural University,30(3):1-8.(in Chinese)
[17] 田清震,盖钧镒,喻德跃,等.我国野生大豆与栽培大豆A FLP指纹图谱研究[J].中国农业科学,2001,34(5):465 - 468TIAN Qing-zhen,GAI Jun-yi,YU De-yue,et al.(2001).AFL P fingerprint analysis of G.soja and G.max in China [J].Scientia Agricultura Sinica,34(5):465- 468.(in Chinese)
[18] 王凤格,杨扬,易红梅,等.中国玉米审定品种标准SSR指纹库的构建[J].中国农业科学,2017,50(1):1-14.WANG Feng-ge,YANG Yang,YI Hong-mei,et al.(2017).Construction of an SSR-Based Standard Fingerprint Database for Corn Variety Authorized in China [J].Scientia Agricultura Sinica,50(1):1-14.(in Chinese)
[19] 徐军,刘艳华,任民,等.普通烟草种质资源的 SSR 标记与指纹图谱分析[J].中国烟草科学,2011,32(2):62-65.XU Jun,LIU Yan-hua,REN Min,et al.(2011).SSR fingerprint map analysis of tobacco germplasms [J].Chinese Gobacco Science,32(2):62- 65.(in Chinese)
[20] 张小娟,何团结,陆徐忠,等.陆地棉SSR核心引物筛选及95份骨干种质的遗传多样性分析[J].棉花学报,2011,23(6):529-536.ZHANG Xiao-juan,HE Tuan-jie,LU Xu-zhong,et al.(2011).Screening of the SSR Core Primers of Upland Cotton and Analysis of Genetic Diversity for 95 Core Germplasms with SSR Markers [J].Cotton Science,23(6):529-536.(in Chinese)
[21] 朱美霞,李英芝,王建书,等.利用SSR方法鉴定棉花品种纯度[J].安徽农业科学,2005,33(11):2 010-2 016.ZHU Mei-xia,LI Ying-zhi,WANG Jian-shu,et al.(2005).Purity identification of cotton varieties with SSR technique [J].Journal of Anhui Agicultural Sciences,33(11):2,010-2,016.(in Chinese)