不同品种群牡丹表型SSR标记的遗传多样性及聚类分析
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摘要
为了解牡丹种质资源的多样性,利用22个自主开发的SSR标记对来自5个品种群的30个牡丹种质多态性及其亲缘关系进行了分析。结果表明:22个SSR位点具有多态性,共检测到90个等位基因,各位点等位基因数介于2—6个,平均等位基因数为4.09个,Nei’s基因多样性指数为0.0713—0.7469,Shannon’s多样性指数(I)为0.1584—1.4000,观望杂合度(Ho)、期望杂合度(He)、多态信息含量(PIC)范围分别为0.1481—1.000、0.0727—0.7908、0.0713—0.7469。22个多态位点中有16个高度多态、2个中度多态、4个低度多态。UPGMA聚类分析结果表明:江南品种与西南品种聚为一支,中原品种、西北品种和国际品种聚为一支。有效SSR标记的开发及牡丹品种资源间亲缘关系的揭示,可为牡丹种质资源保存及分子标记辅助育种奠定科学基础。
In order to understand the diversity of peony germplasm resources, 22 self-developed SSRmarkers were applied to analyze 30 tree peony germplasm polymorphisms and phylogenetic relationships from 5 cultivar groups. The results showed that 22 SSR loci were polymorphic and 90 alleles were detected,the allelenumber were ranged from 2 to 6 with an average of 4. 09. Nei.s gene diversity index was 0. 0713—0. 7469,Shannon.s diversity index(I)was 0. 1584—1. 4000,and the range of heterozygosity(Ho),expected heterozygosity(He) and polymorphic information content(PIC) were 0. 1481—1. 000,0. 0727—0. 7908 and 0. 0713—0. 7469,respectively. Of the 22 polymorphic loci,there were 16 high polymorphism,2 moderate polymorphismsand 4 low degree polymorphisms. The results of UPGMA cluster analysis showed that the cultivars of southYangtze River and southwest China were gathered together,while the cultivars of Central Plains,northwest Chinaand international were gathered together. The development of effective SSR markers and the relationship betweenthe resources of peony varieties could lay a scientific foundation for the preservation of tree peony germplasmresources and the molecular marker assisted breeding.
In order to understand the diversity of peony germplasm resources, 22 self-developed SSRmarkers were applied to analyze 30 tree peony germplasm polymorphisms and phylogenetic relationships from 5 cultivar groups. The results showed that 22 SSR loci were polymorphic and 90 alleles were detected,the allelenumber were ranged from 2 to 6 with an average of 4. 09. Nei.s gene diversity index was 0. 0713—0. 7469,Shannon.s diversity index(I)was 0. 1584—1. 4000,and the range of heterozygosity(Ho),expected heterozygosity(He) and polymorphic information content(PIC) were 0. 1481—1. 000,0. 0727—0. 7908 and 0. 0713—0. 7469,respectively. Of the 22 polymorphic loci,there were 16 high polymorphism,2 moderate polymorphismsand 4 low degree polymorphisms. The results of UPGMA cluster analysis showed that the cultivars of southYangtze River and southwest China were gathered together,while the cultivars of Central Plains,northwest Chinaand international were gathered together. The development of effective SSR markers and the relationship betweenthe resources of peony varieties could lay a scientific foundation for the preservation of tree peony germplasmresources and the molecular marker assisted breeding.
引文
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[3]SU J,MA C,LIU C,et al.Hypolipidemic activity of peony seed oil rich in alpha-linolenic,is mediated through inhibition of lipogenesis and upregulation of fatty acid beta-oxidation[J].Journal of Food Science,2016,81(4):1001-1009.
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[8]郭琪,郭大龙,郭丽丽,等.SSR分子标记在牡丹亲缘关系研究中的应用与研究进展[J].植物学报,2015,50(5):652-664.
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[11]GUO D L,HOU X G,ZHANG J.Sequence-related amplified polymorphism analysis of tree peony(Paeonia suffruticosaAndrews)cultivars with different flower colours[J].Journal of Horticultural Science&Biotechnology,2009,84(2):131-136.
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[14]HOU X G,GUO D L,CHENG S P,et al.Development of thirty new polymorphic microsatellite primers forPaeonia suffruticosa[J].Biologia Plantarum,2011,55(4):708-710.
[15]WANG X,FAN H,LI Y,et al.Analysis of genetic relationships in tree peony of different colors using conserved DNA-derived polymorphism markers[J].Scientia Horticulturae,2014,175:68-73.
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[17]李宗艳,秦艳玲,蒙进芳,等.西南牡丹品种起源的ISSR研究[J].中国农业科学,2015,48(5):931-940.
[18]钟原,成仿云,吴静.牡丹(Paeoniasect.Moutan)分子标记研究进展[J].分子植物育种,2016,14(6):1559-1567.
[19]TAUTZ D.Hypervariabflity of simple sequences as a general source for polymorphic DNA markers[J].Nucleic acids research,1989,17(16):6463-6471.
[20]WANG J,XIA T,ZHANG J,et al.Isolation and characterization of fourteen microsatellites from a tree peony(Paeonia suffruticosa)[J].Conservation Genetics,2009,10(4):1029-1031.
[21]HOMOLKA A,BERENYI M,BURG K,et al.Microsatellite markers in the tree peony,Paeonia suffruticosa(Paeoniaceae)[J].American Journal of Botany,2010,97(6):42-44.
[22]GAI S P,ZHANG Y X,MU P,et al.Transcriptome analysis of tree peony during chilling requirement fulfillment:assembling,annotation and markers discovering[J].Gene,2012,497(2):256-262.
[23]WU J,CAI C F,CHENG F Y,et al.Characterisation and development of EST-SSR markers in tree peony using transcriptome sequences[J].Molecular Breeding,2014,34(4):1853-1866.
[24]GAO Z M,WU J,LIU Z A,et al.Rapid microsatellite development for tree peony and its implications[J].BMC Genomics,2013,14(1):886.
[25]YU H P,CHENG F Y,ZHONG Y,et al.Development of simple sequence repeat(SSR)markers fromPaeonia ostiito study the genetic relationships among tree peonies(Paeoniaceae)[J].Scientia Horticulturae,2013,164:58-64.
[26]WU J,CHENG F Y,CAI C F,et al.Association mapping for floral traits in cultivatedPaeonia rockiibased on SSR markers[J].Molecular genetics and genomics,2017,292(1):187-200.
[27]CAI C F,CHENG F Y,WU J,et al.The first high-density genetic map construction in tree peony(PaeoniaSect.Moutan)using genotyping by specific-locus amplified fragment sequencing[J].Plos One,2015,10(5):e0128584.
[28]ZHANG J M,LIU J,SUN H L,et al.Nuclear and chloroplast SSR markers inPaeonia delavayi(paeoniaceae)and cross-species amplification inP.Ludlowii[J].American Journal of Botany,2011,98(12):346-348.
[29]ZHANG J J,SHU Q Y,LIU Z A,et al.Two EST-derived marker systems for cultivar identification in tree peony[J].Plant Cell Reports,2012,31(2):299-310.
[30]WEN S S,CHENG F Y,ZHONG Y,et al.Efficient protocols for the micropropagation of tree peony(Paeonia suffruticosa‘Jin Pao Hong’,P.suffruticosa‘Wu Long Peng Sheng’,andP.×lemoinei‘High Noon’)and application of Arbuscular mycorrhizal fungi to improve plantlet establishment[J].Scientia Horticulturae,2016,201:10-17.
[31]GILMORE B,BASSIL N,NYBERG A,et al.Microsatellite marker development in peony using next generation sequencing[J].Journal of the American Society for Horticultural Science,2013,138(1):64-74.
[32]NEI M.Estimation of average heterozygosity and genetic distance from a small number of individuals[J].Genetics,1978,89(3):583-590.
[33]MOGEA J P.Relationships and phylogeny of the species of the genusArenga(Palmae)based on morphology using the polarity method and the NTSYS program[J].Mem.New York Bot.Gard,1999,83:169-177.
[34]张艳欣,林忠旭,李武,等.海岛棉EST-SSR引物的开发与应用研究[J].科学通报,2007,52(15):1779-1787.
[35]王西成,姜淑苓,上官凌飞,等.梨EST-SSR标记的开发及其在梨品种遗传多样性分析中的应用评价[J].中国农业科学,2010,43(24):5079-5087.
[36]董清华,王西成,赵密珍,等.草莓EST-SSR标记开发及在品种遗传多样性分析中的应用[J].中国农业科学,2011,44(17):3603-3612.
[37]李美芹,潘叶羽,钱萍仙,等.杜鹃花EST-SSR标记的开发及遗传多样性分析[J].植物生理学报,2016,52(3):356-364.
[38]黄磊,王义权.微卫星分子标记在濒危动物保护遗传学研究中的应用[J].生物多样性,2004,12(5):528-533.
[39]ZHAO X,ZHOU Z Q,LIN Q B,et al.Phylogenetic analysis of Paeonia sect.Moutan(Paeoninceae)based on multiple DNA fragments and morphological data[J].Journal of Systematics and Evolution,2008,46(4):563-572.
[40]石颜通,周波,张秀新,等.牡丹89个不同种源品种遗传多样性和亲缘关系分析[J].园艺学报,2012,39(12):2499-2506.
[41]周志钦,潘开玉,洪德元.牡丹组野生种间亲缘关系和栽培牡丹起源研究进展[J].园艺学报,2003,30(6):751-757.
[42]蔡长福.牡丹高密度遗传图谱构建及重要性状QTL分析[D].北京:北京林业大学,2015.
[2]SARKER S D,WHITING P,DINAN L,et al.Identification and ecdysteroid antagonist activity of three resveratrol trimers(suffruticosols A,B and C)fromPaeonia suffruticosa[J].Tetrahedron,1999,55(2):513-524.
[3]SU J,MA C,LIU C,et al.Hypolipidemic activity of peony seed oil rich in alpha-linolenic,is mediated through inhibition of lipogenesis and upregulation of fatty acid beta-oxidation[J].Journal of Food Science,2016,81(4):1001-1009.
[4]ROGERS A:Peonies[M].Portland:Timber Press,1995.
[5]CHENG F.Advances in the breeding of tree peonies and a cultivar system for the cultivar group[J].International Journal of Plant Breeding,2007,1(2):89-104.
[6]李嘉珏,张西方,赵孝庆.中国牡丹[M].北京:中国大百科全书出版社,2011.
[7]郭大龙.牡丹种质资源遗传多样性研究进展[J].北方园艺,2007(9):61-65.
[8]郭琪,郭大龙,郭丽丽,等.SSR分子标记在牡丹亲缘关系研究中的应用与研究进展[J].植物学报,2015,50(5):652-664.
[9]陈向明,郑国生,张圣旺.牡丹栽培品种的RAPD分析[J].园艺学报,2001,28(4):370-372.
[10]HOU X G,YIN W,LI J J,et al.AFLP analysis of genetic diversity of 30 tree peony(Paeonia suffruticosaAndr.)cultivars[J].Scientia Agri Sinica,2006,39:1709-1715.
[11]GUO D L,HOU X G,ZHANG J.Sequence-related amplified polymorphism analysis of tree peony(Paeonia suffruticosaAndrews)cultivars with different flower colours[J].Journal of Horticultural Science&Biotechnology,2009,84(2):131-136.
[12]袁军辉.紫斑牡丹及延安牡丹起源研究[D].北京:北京林业大学,2010.
[13]HOU X G,GUO D L,WANG J.Development and characterization of EST-SSR markers inPaeonia suffruticosa(Paeoniaceae)[J].American Journal of Botany,2011,98(11):303-305.
[14]HOU X G,GUO D L,CHENG S P,et al.Development of thirty new polymorphic microsatellite primers forPaeonia suffruticosa[J].Biologia Plantarum,2011,55(4):708-710.
[15]WANG X,FAN H,LI Y,et al.Analysis of genetic relationships in tree peony of different colors using conserved DNA-derived polymorphism markers[J].Scientia Horticulturae,2014,175:68-73.
[16]ZHOU S L,ZOU X H,ZHOU Z Q,et al.Multiple species of wild tree peonies gave rise to the‘king of flowers’,Paeonia suffruticosaAndrews[J].Proceedings of the Royal Society of London B:Biological Sciences,2014,281(1797):20141687.
[17]李宗艳,秦艳玲,蒙进芳,等.西南牡丹品种起源的ISSR研究[J].中国农业科学,2015,48(5):931-940.
[18]钟原,成仿云,吴静.牡丹(Paeoniasect.Moutan)分子标记研究进展[J].分子植物育种,2016,14(6):1559-1567.
[19]TAUTZ D.Hypervariabflity of simple sequences as a general source for polymorphic DNA markers[J].Nucleic acids research,1989,17(16):6463-6471.
[20]WANG J,XIA T,ZHANG J,et al.Isolation and characterization of fourteen microsatellites from a tree peony(Paeonia suffruticosa)[J].Conservation Genetics,2009,10(4):1029-1031.
[21]HOMOLKA A,BERENYI M,BURG K,et al.Microsatellite markers in the tree peony,Paeonia suffruticosa(Paeoniaceae)[J].American Journal of Botany,2010,97(6):42-44.
[22]GAI S P,ZHANG Y X,MU P,et al.Transcriptome analysis of tree peony during chilling requirement fulfillment:assembling,annotation and markers discovering[J].Gene,2012,497(2):256-262.
[23]WU J,CAI C F,CHENG F Y,et al.Characterisation and development of EST-SSR markers in tree peony using transcriptome sequences[J].Molecular Breeding,2014,34(4):1853-1866.
[24]GAO Z M,WU J,LIU Z A,et al.Rapid microsatellite development for tree peony and its implications[J].BMC Genomics,2013,14(1):886.
[25]YU H P,CHENG F Y,ZHONG Y,et al.Development of simple sequence repeat(SSR)markers fromPaeonia ostiito study the genetic relationships among tree peonies(Paeoniaceae)[J].Scientia Horticulturae,2013,164:58-64.
[26]WU J,CHENG F Y,CAI C F,et al.Association mapping for floral traits in cultivatedPaeonia rockiibased on SSR markers[J].Molecular genetics and genomics,2017,292(1):187-200.
[27]CAI C F,CHENG F Y,WU J,et al.The first high-density genetic map construction in tree peony(PaeoniaSect.Moutan)using genotyping by specific-locus amplified fragment sequencing[J].Plos One,2015,10(5):e0128584.
[28]ZHANG J M,LIU J,SUN H L,et al.Nuclear and chloroplast SSR markers inPaeonia delavayi(paeoniaceae)and cross-species amplification inP.Ludlowii[J].American Journal of Botany,2011,98(12):346-348.
[29]ZHANG J J,SHU Q Y,LIU Z A,et al.Two EST-derived marker systems for cultivar identification in tree peony[J].Plant Cell Reports,2012,31(2):299-310.
[30]WEN S S,CHENG F Y,ZHONG Y,et al.Efficient protocols for the micropropagation of tree peony(Paeonia suffruticosa‘Jin Pao Hong’,P.suffruticosa‘Wu Long Peng Sheng’,andP.×lemoinei‘High Noon’)and application of Arbuscular mycorrhizal fungi to improve plantlet establishment[J].Scientia Horticulturae,2016,201:10-17.
[31]GILMORE B,BASSIL N,NYBERG A,et al.Microsatellite marker development in peony using next generation sequencing[J].Journal of the American Society for Horticultural Science,2013,138(1):64-74.
[32]NEI M.Estimation of average heterozygosity and genetic distance from a small number of individuals[J].Genetics,1978,89(3):583-590.
[33]MOGEA J P.Relationships and phylogeny of the species of the genusArenga(Palmae)based on morphology using the polarity method and the NTSYS program[J].Mem.New York Bot.Gard,1999,83:169-177.
[34]张艳欣,林忠旭,李武,等.海岛棉EST-SSR引物的开发与应用研究[J].科学通报,2007,52(15):1779-1787.
[35]王西成,姜淑苓,上官凌飞,等.梨EST-SSR标记的开发及其在梨品种遗传多样性分析中的应用评价[J].中国农业科学,2010,43(24):5079-5087.
[36]董清华,王西成,赵密珍,等.草莓EST-SSR标记开发及在品种遗传多样性分析中的应用[J].中国农业科学,2011,44(17):3603-3612.
[37]李美芹,潘叶羽,钱萍仙,等.杜鹃花EST-SSR标记的开发及遗传多样性分析[J].植物生理学报,2016,52(3):356-364.
[38]黄磊,王义权.微卫星分子标记在濒危动物保护遗传学研究中的应用[J].生物多样性,2004,12(5):528-533.
[39]ZHAO X,ZHOU Z Q,LIN Q B,et al.Phylogenetic analysis of Paeonia sect.Moutan(Paeoninceae)based on multiple DNA fragments and morphological data[J].Journal of Systematics and Evolution,2008,46(4):563-572.
[40]石颜通,周波,张秀新,等.牡丹89个不同种源品种遗传多样性和亲缘关系分析[J].园艺学报,2012,39(12):2499-2506.
[41]周志钦,潘开玉,洪德元.牡丹组野生种间亲缘关系和栽培牡丹起源研究进展[J].园艺学报,2003,30(6):751-757.
[42]蔡长福.牡丹高密度遗传图谱构建及重要性状QTL分析[D].北京:北京林业大学,2015.