三种海岛棉指纹图谱建立
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摘要
为了筛选陆海杂交后与海岛棉具有相近遗传背景的材料,利用海岛棉做轮回亲本,通过回交使杂交后代背景与海岛棉一致,将具有供体陆地棉的目标性状,分别构建陆地棉和海岛棉的指纹图谱,利用指纹图谱进行杂交后代遗传背景选择。选取供体陆地棉与三种受体海岛棉作为实验材料,采用分子标记技术,对实验室已有的1 300对SSR引物进行筛选,对扩增条带清晰、重复性好且分辨能力强的核心引物进行统计分析,确定不同棉花材料的指纹图谱。筛选出63对陆地棉和海岛棉之间差异性大、多态性好的引物,分别分布于棉花26条染色体,并分别构建了供体陆地棉和三种海岛棉的指纹图谱。筛选出的这63对引物具有良好的鉴别和区分能力,适用于作为首选的核心引物。本研究为开展海岛棉与陆地棉杂交后利用回交快速获得纯合遗传背景的杂交后代提供了研究思路。
In order to screen materials with similar genetic background to island cotton after land-sea hybridization,island cotton was used as recurrent parent to make the background of hybrid progeny consistent with island cotton,but with the target traits of donor upland cotton. The fingerprints of upland cotton and sea island cotton were constructed and used to select the genetic background of hybrids. Donor upland cotton and three kinds of receptors sea island cotton were selected as experiment material, and 1 300 pairs of primers in the laboratory were screened by molecular marker technology. The core primers with clear amplified bands, good repeatability and strong resolution were analyzed statistically to determine the fingerprints of different cotton materials. 63 pairs of primers with good diversity and polymorphism were screened out from upland cotton and sea island cotton, which were distributed on 26 chromosomes of cotton. In addition, the fingerprint of donor upland cotton and three kinds of sea island cotton were constructed. The selected 63 pairs of primers had good identification and distinguishing ability,which were suitable as the preferred core primer. This study could provide a research idea for the rapid generation of homozygous genetic background by backcross between sea island cotton and upland cotton.
In order to screen materials with similar genetic background to island cotton after land-sea hybridization,island cotton was used as recurrent parent to make the background of hybrid progeny consistent with island cotton,but with the target traits of donor upland cotton. The fingerprints of upland cotton and sea island cotton were constructed and used to select the genetic background of hybrids. Donor upland cotton and three kinds of receptors sea island cotton were selected as experiment material, and 1 300 pairs of primers in the laboratory were screened by molecular marker technology. The core primers with clear amplified bands, good repeatability and strong resolution were analyzed statistically to determine the fingerprints of different cotton materials. 63 pairs of primers with good diversity and polymorphism were screened out from upland cotton and sea island cotton, which were distributed on 26 chromosomes of cotton. In addition, the fingerprint of donor upland cotton and three kinds of sea island cotton were constructed. The selected 63 pairs of primers had good identification and distinguishing ability,which were suitable as the preferred core primer. This study could provide a research idea for the rapid generation of homozygous genetic background by backcross between sea island cotton and upland cotton.
引文
Chen H.D.,Xiao C.S.,Li Y.,and Kuang Z.C.,Wang W.,and Li Y.Q.,2014,DNA fingerprinting and genetic diversity analysis of short season cotton by SSR markers,Huabei Nongxue Bao(Acta Agriculturae Boreali-Sinica),29(4):98-104(陈浩东,肖才升,李庠,匡政成,王为,李育强,2014,基于SSR标记的短季棉品种DNA指纹图谱构建及遗传多样性分析,华北农学报,29(4):98-104)
Chen X.H.,Xu L.Y.,Peng Z.S.,Du W.P.,Yu G.R.,Zhong C.S.,Qu J.P.,and Hu F.L.,2008,SSR analysis of genetic diversity for wheat cultivars(lines)in the southwest of China,Mailei Zuowu Xuebao(Journal of Triticeae Crops),28(1):6-10(陈先红,徐利远,彭正松,杜文平,余桂蓉,钟昌松,曲继鹏,胡凤林,2008,中国西南地区小麦品种(系)遗传多样性的SSR分析,麦类作物学报,28(1):6-10)
Deng C.L.,Gao J.,Cao Y.,Qin R.Y.,Gao W.J.,and Lu L.D.,2012,Development and application of the sex-specific EST-SSRmarker inspinach(Spinacia oleracea L.),Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),31(5):467-472(邓传良,高俊,曹莹,秦瑞云,高武军,卢龙斗,2012,菠菜性别相关EST-SSR标记的开发及应用,基因组学与应用生物学,31(5):467-472)
Hao C.Y.,Wang L.F.,Jia J.Z.,Dong Y.C.,and Zhang X.Y.,2005,Comparison of fluorescence and silver-staining detection systems of microsatellite markers,Zuowu Xuebao(Acta A-gronomica Sinica),31(2):144-149(郝晨阳,王兰芬,贾继增,董玉琛,张学勇,2005,SSR荧光标记和银染技术的比较分析,作物学报,31(2):144-149)
Hua L.,Yuan Y.P.,Yu H.Y.,Wang Y.P.,Xu Q.,Tang S.X.,and Wei X.H.,2007,A comparative study on SSR diversity in Chinese major rice varieties planted in 1950s and during the most recent ten years,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),21(2):150-154(华蕾,袁筱萍,余汉勇,王一平,徐群,汤圣祥,魏兴华,2007,我国水稻主栽品种SSR多样性的比较分析,中国水稻科学,21(2):150-154)
Is Iam M.N.,Molla M.K.,Mm R.,Hasanuzzaman M.,Islam S.M.N.,and Rahman L.,2012,DNA fingerprinting and genotyping of cotton varieties using SSR markers,Not.Bot.Horti Agrobo.,40(2):261-265
Lacape J.M.,Dessauw D.,Rajab M.,Noyer J.L.,and Hau B.,2007,Microsatellite diversity in tetraploid Gossypium germplasm:assembling a highly informative genotyping set of cotton SSRs,Mol.Breed.,19(1):45-58
Li H.B.,Yang J.,LüZ.W.,Yi B.,Wen J.,Bo T.D.,Tu J.X.,Ma C.Z.,and Shen J.X.,2010,Screening of Brassica napus core SSR priners,Zhongguo Youliao Zuowu Xuebao(Chinese Journal of Oil Crop Sciences),32(3):329-336(李海渤,杨军,吕泽文,易斌,文静,傅廷栋,涂金星,马朝芝,沈金雄,2010,甘蓝型油菜SSR核心引物研究,中国油料作物学报,32(3):329-336)
Li L.,Wang J.F.,Yan T.J.,Li N.N.,Ding H.J.,and Fan S.J.,2013,Eetablishiment of DNA fingerprint of wheat in Shandong Province based on SSR markers,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),14(3):537-541(李莉,王俊峰,颜廷进,李娜娜,丁汉金,樊守金,基于SSR标记的山东省小麦DNA指纹图谱的构建,植物遗传资源学报,14(3):537-541)
Liu F.,Feng X.M.,Zhong W.,Liu Y.D.,Yin Z.J.,Han X.L.,Xu Z.C.,and Shen F.F.,2009,Screening of SSR core primer pairs for identifying cotton cultivar,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),7(6):1160-1168(刘峰,冯雪梅,钟文,刘玉栋,阴祖军,韩秀兰,徐子初,沈法富,2009,适合棉花品种鉴定的SSR核心引物的筛选,分子植物育种,7(6):1160-1168)
Liu G.D.,Wang F.R.,Gong Y.C.,Ma H.H.,and Zhang J.,2013,A new method for identification of the genetic purity of cotton varieties by SSR markers,Mianhua Xuebao(Cotton Science),25(5):382-387(刘国栋,王芙蓉,宫永超,马和欢,张军,2013,棉花品种遗传纯度的SSR分子标记鉴定技术研究,棉花学报,25(5):382-387)
Pan Z.E.,Sun J.L.,Wang X.W.,Jia Y.H.,Zhou Z.L.,Pang B.Y.,and Du X.M.,2008,Screening of SSR core primers with polymorphism on a cotton panel,Shengwu Duoyangxing(Biodiversity Science),16(6):555-561(潘兆娥,孙君灵,王西文,贾银华,周忠丽,庞保印,杜雄明,2008,基于棉花参比种质的SSR多态性核心引物筛选,生物多样性,16(6):555-561)
Porebski S.,Bailey L.G.,and Baum B.R.,1997,Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components,Plant Mol.Biol.Rep.,15(1):8-15
Qin Y.S.,Liu R.C.,Mei H.X.,Zhang T.Z.,and Guo W.Z.,2009,QTL mapping for yield traits in upland cotton(Gossypium hirsutum L.),Zuowu Xuebao(Acta Agronomica Sinica),35(10):1812-1821(秦永生,刘任重,梅鸿献,张天真,郭旺珍,2009,陆地棉产量相关性状的QTL定位,作物学报,35(10):1812-1821)
Tang S.Y.,Teng Z.H.,Zhai T.F.,Fang X.M.,Liu F.,Liu D.J.,Zhang J.,Liu D.X.,Wang S.F.,Zhang K.,Shao Q.S.,Tan Z.Y.,Paterson A.H.,and Zhang Z.S.,2015,Construction of genetic map and QTL analysis of fiber quality traits for upland cotton(Cossypium hirsutum L.),Euphytica,201(2):195-213
Tian D.G.,Lin Y.,Liu H.Q.,Su J.,Chen Z.J.,Yan J.W.,Xu Y.,and Wang F.,2013,Fingerprinting analysis of 123 rice varieties based on the new SSR core markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),11(1):20-29(田大刚,林艳,刘华清,苏军,陈在杰,颜静宛,许彦,王锋,2013,123份水稻重要品种的SSR核心标记指纹分析,分子植物育种,11(1):20-29)
Wang X.W.,2009,Genetic diversity evaluation and establishment of SSR digital fingerprint of the elite germplasm resource in cotton,Thesis for M.S.,Sichuan Agricultural U-niersity,Supervisors:Feng Z.Y.,and Du X.M.,pp.49-50(王希文,2009,棉花优异种质遗传多样性分析及其SSR数字指纹图谱构建,硕士学位论文,四川农业大学,导师:冯宗云,杜雄明,pp.49-50)
Yao Q.,Yang K.,Pan G.,and Rong T.,2007,Genetic diversity of maize(Zea mays L.)landraces from Southwest China based on SSR data,J.Genet.Genomics,34(9):851-860
You C.Y.,Nie X.H.,Zhang S.,Guo H.L.,Wang X.Q.,and Lin Z.X.,2014,Construction of DNA fingerprinting and analysis of genetic diversity with SSR markers for 23 color cotton cultivars in Xinjiang,Mianhua Xuebao(Cotton Science),26(2):161-170(尤春源,聂新辉,张胜,郭欢乐,王夏青,林忠旭,2014,新疆彩色棉23个品种指纹图谱的构建及遗传多样性分析,棉花学报,26(2):161-170)
Zhang T.Z.,Yuan Y.L.,Guo W.Z.,Yu J.,and Kohel R.J.,2001,Microsatellite marker screening for high strength fiber QTLs of cotton,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),34(4):363-366(张天真,袁有禄,郭旺珍,Yu J.,Kohel R.J.,2001,棉花高强纤维QTLs的微卫星标记筛选,中国农业科学,34(4):363-366)
Zhu Y.J.,Fan Y.Y.,Huang D.R.,and Zhuang J.Y.,2012,Application of marker issisted selection in rice breeding,Henongxue Bao(Journal of Nuclear Agricultural Sciences),26(5):756-761(朱玉君,樊叶杨,黄得润,庄杰云,2012,分子标记辅助选择技术在水稻育种中的应用,核农学报,26(5):756-761)
Chen X.H.,Xu L.Y.,Peng Z.S.,Du W.P.,Yu G.R.,Zhong C.S.,Qu J.P.,and Hu F.L.,2008,SSR analysis of genetic diversity for wheat cultivars(lines)in the southwest of China,Mailei Zuowu Xuebao(Journal of Triticeae Crops),28(1):6-10(陈先红,徐利远,彭正松,杜文平,余桂蓉,钟昌松,曲继鹏,胡凤林,2008,中国西南地区小麦品种(系)遗传多样性的SSR分析,麦类作物学报,28(1):6-10)
Deng C.L.,Gao J.,Cao Y.,Qin R.Y.,Gao W.J.,and Lu L.D.,2012,Development and application of the sex-specific EST-SSRmarker inspinach(Spinacia oleracea L.),Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),31(5):467-472(邓传良,高俊,曹莹,秦瑞云,高武军,卢龙斗,2012,菠菜性别相关EST-SSR标记的开发及应用,基因组学与应用生物学,31(5):467-472)
Hao C.Y.,Wang L.F.,Jia J.Z.,Dong Y.C.,and Zhang X.Y.,2005,Comparison of fluorescence and silver-staining detection systems of microsatellite markers,Zuowu Xuebao(Acta A-gronomica Sinica),31(2):144-149(郝晨阳,王兰芬,贾继增,董玉琛,张学勇,2005,SSR荧光标记和银染技术的比较分析,作物学报,31(2):144-149)
Hua L.,Yuan Y.P.,Yu H.Y.,Wang Y.P.,Xu Q.,Tang S.X.,and Wei X.H.,2007,A comparative study on SSR diversity in Chinese major rice varieties planted in 1950s and during the most recent ten years,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),21(2):150-154(华蕾,袁筱萍,余汉勇,王一平,徐群,汤圣祥,魏兴华,2007,我国水稻主栽品种SSR多样性的比较分析,中国水稻科学,21(2):150-154)
Is Iam M.N.,Molla M.K.,Mm R.,Hasanuzzaman M.,Islam S.M.N.,and Rahman L.,2012,DNA fingerprinting and genotyping of cotton varieties using SSR markers,Not.Bot.Horti Agrobo.,40(2):261-265
Lacape J.M.,Dessauw D.,Rajab M.,Noyer J.L.,and Hau B.,2007,Microsatellite diversity in tetraploid Gossypium germplasm:assembling a highly informative genotyping set of cotton SSRs,Mol.Breed.,19(1):45-58
Li H.B.,Yang J.,LüZ.W.,Yi B.,Wen J.,Bo T.D.,Tu J.X.,Ma C.Z.,and Shen J.X.,2010,Screening of Brassica napus core SSR priners,Zhongguo Youliao Zuowu Xuebao(Chinese Journal of Oil Crop Sciences),32(3):329-336(李海渤,杨军,吕泽文,易斌,文静,傅廷栋,涂金星,马朝芝,沈金雄,2010,甘蓝型油菜SSR核心引物研究,中国油料作物学报,32(3):329-336)
Li L.,Wang J.F.,Yan T.J.,Li N.N.,Ding H.J.,and Fan S.J.,2013,Eetablishiment of DNA fingerprint of wheat in Shandong Province based on SSR markers,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),14(3):537-541(李莉,王俊峰,颜廷进,李娜娜,丁汉金,樊守金,基于SSR标记的山东省小麦DNA指纹图谱的构建,植物遗传资源学报,14(3):537-541)
Liu F.,Feng X.M.,Zhong W.,Liu Y.D.,Yin Z.J.,Han X.L.,Xu Z.C.,and Shen F.F.,2009,Screening of SSR core primer pairs for identifying cotton cultivar,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),7(6):1160-1168(刘峰,冯雪梅,钟文,刘玉栋,阴祖军,韩秀兰,徐子初,沈法富,2009,适合棉花品种鉴定的SSR核心引物的筛选,分子植物育种,7(6):1160-1168)
Liu G.D.,Wang F.R.,Gong Y.C.,Ma H.H.,and Zhang J.,2013,A new method for identification of the genetic purity of cotton varieties by SSR markers,Mianhua Xuebao(Cotton Science),25(5):382-387(刘国栋,王芙蓉,宫永超,马和欢,张军,2013,棉花品种遗传纯度的SSR分子标记鉴定技术研究,棉花学报,25(5):382-387)
Pan Z.E.,Sun J.L.,Wang X.W.,Jia Y.H.,Zhou Z.L.,Pang B.Y.,and Du X.M.,2008,Screening of SSR core primers with polymorphism on a cotton panel,Shengwu Duoyangxing(Biodiversity Science),16(6):555-561(潘兆娥,孙君灵,王西文,贾银华,周忠丽,庞保印,杜雄明,2008,基于棉花参比种质的SSR多态性核心引物筛选,生物多样性,16(6):555-561)
Porebski S.,Bailey L.G.,and Baum B.R.,1997,Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components,Plant Mol.Biol.Rep.,15(1):8-15
Qin Y.S.,Liu R.C.,Mei H.X.,Zhang T.Z.,and Guo W.Z.,2009,QTL mapping for yield traits in upland cotton(Gossypium hirsutum L.),Zuowu Xuebao(Acta Agronomica Sinica),35(10):1812-1821(秦永生,刘任重,梅鸿献,张天真,郭旺珍,2009,陆地棉产量相关性状的QTL定位,作物学报,35(10):1812-1821)
Tang S.Y.,Teng Z.H.,Zhai T.F.,Fang X.M.,Liu F.,Liu D.J.,Zhang J.,Liu D.X.,Wang S.F.,Zhang K.,Shao Q.S.,Tan Z.Y.,Paterson A.H.,and Zhang Z.S.,2015,Construction of genetic map and QTL analysis of fiber quality traits for upland cotton(Cossypium hirsutum L.),Euphytica,201(2):195-213
Tian D.G.,Lin Y.,Liu H.Q.,Su J.,Chen Z.J.,Yan J.W.,Xu Y.,and Wang F.,2013,Fingerprinting analysis of 123 rice varieties based on the new SSR core markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),11(1):20-29(田大刚,林艳,刘华清,苏军,陈在杰,颜静宛,许彦,王锋,2013,123份水稻重要品种的SSR核心标记指纹分析,分子植物育种,11(1):20-29)
Wang X.W.,2009,Genetic diversity evaluation and establishment of SSR digital fingerprint of the elite germplasm resource in cotton,Thesis for M.S.,Sichuan Agricultural U-niersity,Supervisors:Feng Z.Y.,and Du X.M.,pp.49-50(王希文,2009,棉花优异种质遗传多样性分析及其SSR数字指纹图谱构建,硕士学位论文,四川农业大学,导师:冯宗云,杜雄明,pp.49-50)
Yao Q.,Yang K.,Pan G.,and Rong T.,2007,Genetic diversity of maize(Zea mays L.)landraces from Southwest China based on SSR data,J.Genet.Genomics,34(9):851-860
You C.Y.,Nie X.H.,Zhang S.,Guo H.L.,Wang X.Q.,and Lin Z.X.,2014,Construction of DNA fingerprinting and analysis of genetic diversity with SSR markers for 23 color cotton cultivars in Xinjiang,Mianhua Xuebao(Cotton Science),26(2):161-170(尤春源,聂新辉,张胜,郭欢乐,王夏青,林忠旭,2014,新疆彩色棉23个品种指纹图谱的构建及遗传多样性分析,棉花学报,26(2):161-170)
Zhang T.Z.,Yuan Y.L.,Guo W.Z.,Yu J.,and Kohel R.J.,2001,Microsatellite marker screening for high strength fiber QTLs of cotton,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),34(4):363-366(张天真,袁有禄,郭旺珍,Yu J.,Kohel R.J.,2001,棉花高强纤维QTLs的微卫星标记筛选,中国农业科学,34(4):363-366)
Zhu Y.J.,Fan Y.Y.,Huang D.R.,and Zhuang J.Y.,2012,Application of marker issisted selection in rice breeding,Henongxue Bao(Journal of Nuclear Agricultural Sciences),26(5):756-761(朱玉君,樊叶杨,黄得润,庄杰云,2012,分子标记辅助选择技术在水稻育种中的应用,核农学报,26(5):756-761)