菠菜种质资源SSR指纹图谱构建和遗传多样性分析
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摘要
为研究菠菜品种间的亲缘关系,本研究利用PAGE电泳法对33份菠菜种质资源进行指纹图谱构建和遗传多样性分析。结果表明,筛选出的27对多态性明显、带型稳定的引物共扩增出109个多态性位点,平均每对引物3.7个。经多态性位点分析,最后仅用7对高多态性引物构建了能够区分33份菠菜种质的指纹图谱,为供试材料的鉴别提供参考。聚类分析显示,遗传相似系数变幅为0.58～0.96,在遗传相似系数0.68处可以将所有供试材料分为六类,且分类结果与地理来源相近。此研究结果说明33份菠菜种质具有丰富的遗传多样性,可为菠菜品种选育提供理论依据。
In order to study the genetic relationship among spinach varieties, PAGE electrophoresis was used to construct fingerprints and analyze genetic diversity of 33 spinach germplasm resources in this study. The results showed that a total of 109 polymorphic loci were amplified by 27 pairs of primers with obvious polymorphism and stable band type, with an average of 3.7 primers per pair. After polymorphic locus analysis, the fingerprints of 33 spinach germplasms were constructed with only 7 pairs of high polymorphic primers to provide reference for identification of the tested materials. The cluster analysis indicated that the genetic similarity coefficient was0.58～0.96, and the materials could be divided into 6 groups at the genetic distance of 0.68. The classification results were consistent with geographical origin. The results of this study revealed that 33 spinach germplasms had abundant genetic diversity, which could provide theoretical basis for spinach variety breeding.
In order to study the genetic relationship among spinach varieties, PAGE electrophoresis was used to construct fingerprints and analyze genetic diversity of 33 spinach germplasm resources in this study. The results showed that a total of 109 polymorphic loci were amplified by 27 pairs of primers with obvious polymorphism and stable band type, with an average of 3.7 primers per pair. After polymorphic locus analysis, the fingerprints of 33 spinach germplasms were constructed with only 7 pairs of high polymorphic primers to provide reference for identification of the tested materials. The cluster analysis indicated that the genetic similarity coefficient was0.58～0.96, and the materials could be divided into 6 groups at the genetic distance of 0.68. The classification results were consistent with geographical origin. The results of this study revealed that 33 spinach germplasms had abundant genetic diversity, which could provide theoretical basis for spinach variety breeding.
引文
Ana I.L.,Jack S.,Nurit K.,Nurit K.,and María L.G.,2002,Estimation of between and within accession variation in selected Spanish melon germplasm using RAPD and SSR markers to assess strategies for large collection evaluation,Euphytica,127(1):41-51
Cantini C.,Iezzoni A.F.,Lamboy W.F.,Boritzki M.,and Struss D.,2001,DNA fingerprinting of tetraploid cherry germplasm using simple sequence repeats,J.Am.Soc.Hortic.Sci.,126(2):205-209
Deng C.L.,Gao J.,Cao Y.,Qin R.Y.,Gao W.J.,and Lu L.D.,2012,Development and application of sex-specific EST-SSR marker in spinach(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)
Groben R.,and Wricke G.,2010,Occurrence of microsatellites in spinach sequences from computer databases and development of polymorphic SSR markers,Plant Breeding,117(3):271-274
Hu J.G.,Mou B.Q.,and Brady A.V.,2007,Genetic diversity of38 spinach(Spinacia oleracea L.)germplasm accessions and10 commercial hybrids assessed by TRAP markers,Genet.Resour.Crop Evol.,54(8):1667-1674
Jabar Z.K.K.,Anna M.T.,and Sven B.A.,2006,A genetic linkage map of Spinacia oleracea and localization of a sex determination locus,Euphytica,148(3):311-318
Jia H.,Wei X.C.,Yao Q.J.,Yuan Y.X.,Wang Z.Y.,Jiang J.,Yang S.J.,Zhao Y.Y.,Wang B.,and Zhang X.W.,2017,Genetic diversity analysis of capsicum genus germplasm resources using SSR markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(1):353-363(贾豪,魏小春,姚秋菊,原玉香,王志勇,姜俊,杨双娟,赵艳艳,王彬,张晓伟,2017,辣椒SSR标记种质资源遗传多样性的分析,分子植物育种,15(1):353-363)
Li G.G.,Zheng Y.S.,Li X.Y.,Huang H.D.,and Zhang H.,2016,Genetic diversity of bitter gourd germplasm resources revealed by SSR analysis,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(7):1914-1922(李光光,郑岩松,李向阳,黄红弟,张华,2016,苦瓜种质资源SSR遗传多样性分析,分子植物育种,14(7):1914-1922)
Ma H.B.,Xu X.M.,Wei X.Y.,Yang W.X.,and Zou W.G.,2010,DNA fingerpringing and genetic analysis based on SSRmarkers for rice cultivars in Fujian,Fujian Nongye Xuebao(Fujian Journal of Agricultural Science),25(1):33-38(马红勃,许旭明,韦新宇,杨旺兴,邹文广,2010,基于SSR标记的福建省若干水稻DNA指纹图谱构建及遗传多样性分析,福建农业学报,25(1):33-38)
Mei Y.,Cui Y.L.,Guo J.,Chen H.L.,and Meng S.C.,2014,The genetic diversity of breeding materials of spinach,Hubei Nongye Kexue(Hubei Agricultural Sciences),11:2561-2566(梅燚,崔彦玲,郭军,陈海丽,孟淑春,2014,菠菜育种材料的遗传多样性分析,湖北农业科学,11:2561-2566)
Meng S.C.,2017,Morphological comparison of friut,seeds and genetic diversity analysis of Spinacia oleracea L.Dissertation for Ph.D.,China Agricultural University,Supervisor:Zhang Z.X.,pp.1-72(孟淑春,2017,菠菜果实和种子形态比较及遗传多样性研究,博士学位论文,中国农业大学,导师:张振贤,pp.1-72)
Qian Z.W.,Chen H.L.,and Cui Y.L.,2016,Analysis of the SSRloci and development of molecular markers in Spinacia oleracea transcriptome,Nongye Shengwu Jishu Xubao(Journal of Agricultural Biotechnology),24(11):1688-1697(潜宗伟,陈海丽,崔彦玲,2016,菠菜转录组SSR位点分析及其分子标记的开发,农业生物技术学报,24(11):1688-1697)
Rogers S.O.,and Bendich A.J.,1985,Extraction of DNA from milligram amounts of fresh,herbarium and mummified plant tissues,Plant Mol.Biol.,5(2):69-76
Shi J.L.,Chen X.Z.,Wang K.L.,Huang Z.A.,and Zhu L.J.,2017,Genetic diversity analysis of south China ecotypic cucumber germplasm by SSR markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(6):2440-2449(史建磊,陈先知,王克磊,黄宗安,朱隆静,2017,华南型黄瓜种质遗传多样性的SSR分析,分子植物育种,15(6):2440-2449)
Song H.B.,Cui X.B.,Ma H.Y.,Zhu Z.C.,and Luan F.S.,2012,Construction of DNA fingerprint database based on SSRmarker for varieties(Lines)of Cucumis melo L.,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),45(13):2676-2689(宋海斌,崔喜波,马鸿艳,朱子成,栾非时,2012,基于SSR标记的甜瓜品种(系)DNA指纹图谱库的构建,中国农业科学,45(13):2676-2689)
Su Y.J.,Wang J.,Dai X.B.,Tang J.,Zhao D.L.,Zhang A.,Zhou Z.L.,and Cao Q.H.,2018,Genetic diversity and population structure analysis of 303 sweetpotato landraces using SSRmarkers,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),19(2):243-251(苏一钧,王娇,戴习彬,唐君,赵冬兰,张安,周志林,曹清河,2018,303份甘薯地方种SSR遗传多样性与群体结构分析,植物遗传资源学报,19(2):243-251)
Tang J.L.,Huang C.H.,Wu H.,Lang B.B.,Qu X.Y.,and Xu X.B.,2014,Genetic diversity of wild Actinidia eriantha germplasm based on fruit traits and SSR markers,Yuanyi Xuebao(Acta Horticulturae Sinica),41(6):1198-1206(汤佳乐,黄春辉,吴寒,郎彬彬,曲雪艳,徐小彪,2014,野生毛花猕猴桃果实表型性状及SSR遗传多样性分析,园艺学报,41(6):1198-1206)
Wang J.Y.,Chen Y.Y.,Huang B.Z.,Yu F.,and Wu Y.T.,2009,Establishment of fingerprinting for bananas(Musa nana)by SSR marker,Guoshu Xuebao(Journal of Fruit Science),26(5):733-738(王静毅,陈业渊,黄秉智,于飞,武耀廷,2009,部分香蕉品种SSR指纹图谱的构建,果树学报,26(5):733-738)
Wang M.R.,Xu Y.,Zhan Y.L.,Guo S.G.,Ren Y.,Gong G.Y.,and Zhang H.Y.,2010,Analysis of genentic diversity and population structure in melon(Cucumis melo L.)breeding materials by SSR markers,Huabei Nongxuebao(Acta Agriculturae Boreali-Sinica),25(S):41-46(王美荣,许勇,詹永乐,郭绍贵,任毅,宫国义,张海英,2010,甜瓜育种亲本材料遗传多样性及群体结构的SSR标记分析,华北农学报,25(S):41-46)
Wang X.X.,Zhou Q.,Tao Y.Y.,Xu M.,Yu W.W.,and Xu L.A.,2017,The fingerprints construction and genetic diversity analysis of 48 fruit-used Ginkgo cultivars based on SSRmarkers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(5):1963-1970(王星星,周琦,陶园园,胥猛,郁万文,徐立安,2017,48个果用银杏品种SSR指纹图谱构建与遗传多样性分析,分子植物育种,15(5):1963-1970)
Wen S.N.,Zhong C.L.,Jiang Q.B.,Zhang Y.,Chen Z.,and Chen Y.,2017,Genetic diversity analysis of Manglietia conifera dandy germplasm by SSR markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(9):3788-3797(文珊娜,仲崇禄,姜清彬,张勇,陈珍,陈羽,2017,灰木莲种质资源遗传多样性的SSR分析,分子植物育种,15(9):3788-3797)
Wu Y.N.,Kang J.G.,Wang W.K.,and Meng S.C.,2013,Genetic diversity and relationship of spinach germplasm revealed by AFLPs,Yuanyi Xuebao(Acta Horticulturae Sinica),40(5):913-923(吴娅妮,康俊根,王文科,孟淑春,2013,菠菜种质遗传多样性和亲缘关系的AFLP分析,园艺学报,40(5):913-923)
Zhang N.,2007,Study on the genetic diversity and the low temperature tolerance of spinach germplasm,Thesis for M.S.,Northeast Agricultural University,Supervisor:Qin Z.W.,pp.1-81(张南,2007,菠菜种质资源遗传多样性及耐寒性鉴定,硕士学位论文,东北农业大学,导师:秦智伟,pp.1-81)
Cantini C.,Iezzoni A.F.,Lamboy W.F.,Boritzki M.,and Struss D.,2001,DNA fingerprinting of tetraploid cherry germplasm using simple sequence repeats,J.Am.Soc.Hortic.Sci.,126(2):205-209
Deng C.L.,Gao J.,Cao Y.,Qin R.Y.,Gao W.J.,and Lu L.D.,2012,Development and application of sex-specific EST-SSR marker in spinach(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)
Groben R.,and Wricke G.,2010,Occurrence of microsatellites in spinach sequences from computer databases and development of polymorphic SSR markers,Plant Breeding,117(3):271-274
Hu J.G.,Mou B.Q.,and Brady A.V.,2007,Genetic diversity of38 spinach(Spinacia oleracea L.)germplasm accessions and10 commercial hybrids assessed by TRAP markers,Genet.Resour.Crop Evol.,54(8):1667-1674
Jabar Z.K.K.,Anna M.T.,and Sven B.A.,2006,A genetic linkage map of Spinacia oleracea and localization of a sex determination locus,Euphytica,148(3):311-318
Jia H.,Wei X.C.,Yao Q.J.,Yuan Y.X.,Wang Z.Y.,Jiang J.,Yang S.J.,Zhao Y.Y.,Wang B.,and Zhang X.W.,2017,Genetic diversity analysis of capsicum genus germplasm resources using SSR markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(1):353-363(贾豪,魏小春,姚秋菊,原玉香,王志勇,姜俊,杨双娟,赵艳艳,王彬,张晓伟,2017,辣椒SSR标记种质资源遗传多样性的分析,分子植物育种,15(1):353-363)
Li G.G.,Zheng Y.S.,Li X.Y.,Huang H.D.,and Zhang H.,2016,Genetic diversity of bitter gourd germplasm resources revealed by SSR analysis,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(7):1914-1922(李光光,郑岩松,李向阳,黄红弟,张华,2016,苦瓜种质资源SSR遗传多样性分析,分子植物育种,14(7):1914-1922)
Ma H.B.,Xu X.M.,Wei X.Y.,Yang W.X.,and Zou W.G.,2010,DNA fingerpringing and genetic analysis based on SSRmarkers for rice cultivars in Fujian,Fujian Nongye Xuebao(Fujian Journal of Agricultural Science),25(1):33-38(马红勃,许旭明,韦新宇,杨旺兴,邹文广,2010,基于SSR标记的福建省若干水稻DNA指纹图谱构建及遗传多样性分析,福建农业学报,25(1):33-38)
Mei Y.,Cui Y.L.,Guo J.,Chen H.L.,and Meng S.C.,2014,The genetic diversity of breeding materials of spinach,Hubei Nongye Kexue(Hubei Agricultural Sciences),11:2561-2566(梅燚,崔彦玲,郭军,陈海丽,孟淑春,2014,菠菜育种材料的遗传多样性分析,湖北农业科学,11:2561-2566)
Meng S.C.,2017,Morphological comparison of friut,seeds and genetic diversity analysis of Spinacia oleracea L.Dissertation for Ph.D.,China Agricultural University,Supervisor:Zhang Z.X.,pp.1-72(孟淑春,2017,菠菜果实和种子形态比较及遗传多样性研究,博士学位论文,中国农业大学,导师:张振贤,pp.1-72)
Qian Z.W.,Chen H.L.,and Cui Y.L.,2016,Analysis of the SSRloci and development of molecular markers in Spinacia oleracea transcriptome,Nongye Shengwu Jishu Xubao(Journal of Agricultural Biotechnology),24(11):1688-1697(潜宗伟,陈海丽,崔彦玲,2016,菠菜转录组SSR位点分析及其分子标记的开发,农业生物技术学报,24(11):1688-1697)
Rogers S.O.,and Bendich A.J.,1985,Extraction of DNA from milligram amounts of fresh,herbarium and mummified plant tissues,Plant Mol.Biol.,5(2):69-76
Shi J.L.,Chen X.Z.,Wang K.L.,Huang Z.A.,and Zhu L.J.,2017,Genetic diversity analysis of south China ecotypic cucumber germplasm by SSR markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(6):2440-2449(史建磊,陈先知,王克磊,黄宗安,朱隆静,2017,华南型黄瓜种质遗传多样性的SSR分析,分子植物育种,15(6):2440-2449)
Song H.B.,Cui X.B.,Ma H.Y.,Zhu Z.C.,and Luan F.S.,2012,Construction of DNA fingerprint database based on SSRmarker for varieties(Lines)of Cucumis melo L.,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),45(13):2676-2689(宋海斌,崔喜波,马鸿艳,朱子成,栾非时,2012,基于SSR标记的甜瓜品种(系)DNA指纹图谱库的构建,中国农业科学,45(13):2676-2689)
Su Y.J.,Wang J.,Dai X.B.,Tang J.,Zhao D.L.,Zhang A.,Zhou Z.L.,and Cao Q.H.,2018,Genetic diversity and population structure analysis of 303 sweetpotato landraces using SSRmarkers,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),19(2):243-251(苏一钧,王娇,戴习彬,唐君,赵冬兰,张安,周志林,曹清河,2018,303份甘薯地方种SSR遗传多样性与群体结构分析,植物遗传资源学报,19(2):243-251)
Tang J.L.,Huang C.H.,Wu H.,Lang B.B.,Qu X.Y.,and Xu X.B.,2014,Genetic diversity of wild Actinidia eriantha germplasm based on fruit traits and SSR markers,Yuanyi Xuebao(Acta Horticulturae Sinica),41(6):1198-1206(汤佳乐,黄春辉,吴寒,郎彬彬,曲雪艳,徐小彪,2014,野生毛花猕猴桃果实表型性状及SSR遗传多样性分析,园艺学报,41(6):1198-1206)
Wang J.Y.,Chen Y.Y.,Huang B.Z.,Yu F.,and Wu Y.T.,2009,Establishment of fingerprinting for bananas(Musa nana)by SSR marker,Guoshu Xuebao(Journal of Fruit Science),26(5):733-738(王静毅,陈业渊,黄秉智,于飞,武耀廷,2009,部分香蕉品种SSR指纹图谱的构建,果树学报,26(5):733-738)
Wang M.R.,Xu Y.,Zhan Y.L.,Guo S.G.,Ren Y.,Gong G.Y.,and Zhang H.Y.,2010,Analysis of genentic diversity and population structure in melon(Cucumis melo L.)breeding materials by SSR markers,Huabei Nongxuebao(Acta Agriculturae Boreali-Sinica),25(S):41-46(王美荣,许勇,詹永乐,郭绍贵,任毅,宫国义,张海英,2010,甜瓜育种亲本材料遗传多样性及群体结构的SSR标记分析,华北农学报,25(S):41-46)
Wang X.X.,Zhou Q.,Tao Y.Y.,Xu M.,Yu W.W.,and Xu L.A.,2017,The fingerprints construction and genetic diversity analysis of 48 fruit-used Ginkgo cultivars based on SSRmarkers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(5):1963-1970(王星星,周琦,陶园园,胥猛,郁万文,徐立安,2017,48个果用银杏品种SSR指纹图谱构建与遗传多样性分析,分子植物育种,15(5):1963-1970)
Wen S.N.,Zhong C.L.,Jiang Q.B.,Zhang Y.,Chen Z.,and Chen Y.,2017,Genetic diversity analysis of Manglietia conifera dandy germplasm by SSR markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(9):3788-3797(文珊娜,仲崇禄,姜清彬,张勇,陈珍,陈羽,2017,灰木莲种质资源遗传多样性的SSR分析,分子植物育种,15(9):3788-3797)
Wu Y.N.,Kang J.G.,Wang W.K.,and Meng S.C.,2013,Genetic diversity and relationship of spinach germplasm revealed by AFLPs,Yuanyi Xuebao(Acta Horticulturae Sinica),40(5):913-923(吴娅妮,康俊根,王文科,孟淑春,2013,菠菜种质遗传多样性和亲缘关系的AFLP分析,园艺学报,40(5):913-923)
Zhang N.,2007,Study on the genetic diversity and the low temperature tolerance of spinach germplasm,Thesis for M.S.,Northeast Agricultural University,Supervisor:Qin Z.W.,pp.1-81(张南,2007,菠菜种质资源遗传多样性及耐寒性鉴定,硕士学位论文,东北农业大学,导师:秦智伟,pp.1-81)