四川核桃良种SSR指纹图谱构建及遗传多样性分析
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摘要
为构建四川核桃良种指纹图谱,分析遗传多样性,增强品种间的区分能力,该研究利用SSR标记技术,对四川29个核桃良种进行遗传多样性和聚类分析。结果表明:(1)11对SSR引物共检测到121个基因型和80个等位基因,平均每个位点7.273个等位基因和11个基因型。(2)11个位点的平均有效等位基因数为3.644,平均观察杂合度为0.645,平均期望杂合度为0.718,平均香农信息指数为1.518,平均多态信息含量为0.680。(3)采用引物组合法利用引物wga001、wga032和zmz02构建了29个核桃品种的指纹图谱。(4)聚类分析结果表明,29个核桃品种按照品种类型优先聚类,四川本地核桃品种聚类关系与地理来源没有明显的相关性。研究认为,选用的11个SSR标记能够较好地运用于四川核桃品种的遗传多样性研究(PIC>0.5);29个四川核桃品种间亲缘关系较近,遗传基础相对较窄。
In this study,we investigated the fingerprint and genetic diversity of Sichuan elite Juglans regia cultivars to improve the identification ability for cultivars.The SSR markers were applied for genetic diversity and cluster analysis of 29 elite J.regiacultivars.The results showed that:(1)121 genotypes and 80 alleles were identified by the 11 SSR markers,and per primer pair has 7.273 alleles and 11 genotypes.(2)Mean effective allele number(Ne),heterozygosity(Ho),expected heterozygosity(He),shannon's index(I)and polymorphic information contents(PIC)of the 11 SSR markers were 3.644,0.645,0.718,1.518 and 0.680,respectively.(3)The fingerprint of 29 J.regiacultivars were constructed by primer combination method using primers wga001,wga032 and zmz02.(4)Cluster analysis showed that the 29 J.regia cultivars were classified according to the priority of variety type.There is no obvious correlation among cluster relationship and geographical origin of the local J.regiacultivars in Sichuan.The results suggested that the 11 SSR markers can be applied to analyze the genetic diversity of walnut varieties in Sichuan(PIC>0.5);the genetic relationship of 29 Sichuan J.regiacultivars was relatively close,and theirs genetic basis are relatively narrow.
In this study,we investigated the fingerprint and genetic diversity of Sichuan elite Juglans regia cultivars to improve the identification ability for cultivars.The SSR markers were applied for genetic diversity and cluster analysis of 29 elite J.regiacultivars.The results showed that:(1)121 genotypes and 80 alleles were identified by the 11 SSR markers,and per primer pair has 7.273 alleles and 11 genotypes.(2)Mean effective allele number(Ne),heterozygosity(Ho),expected heterozygosity(He),shannon's index(I)and polymorphic information contents(PIC)of the 11 SSR markers were 3.644,0.645,0.718,1.518 and 0.680,respectively.(3)The fingerprint of 29 J.regiacultivars were constructed by primer combination method using primers wga001,wga032 and zmz02.(4)Cluster analysis showed that the 29 J.regia cultivars were classified according to the priority of variety type.There is no obvious correlation among cluster relationship and geographical origin of the local J.regiacultivars in Sichuan.The results suggested that the 11 SSR markers can be applied to analyze the genetic diversity of walnut varieties in Sichuan(PIC>0.5);the genetic relationship of 29 Sichuan J.regiacultivars was relatively close,and theirs genetic basis are relatively narrow.
引文
[1]田荣欢,刘迪秋,陈朝银,等.核桃分子生物学研究进展[J].生物技术通报,2010,(11):19-24.TIAN R H,LIU D Q,CHEN C Y,et al.Research on molecular biology of walnuts(Juglans)[J].Biotechnology Bulletin,2010,(11):19-24.
[2]肖千文,周兰英,胡庭兴,等.早实核桃新品种‘川早2号’[J].园艺学报,2012,39(11):2 317-2 318.XIAO Q W,ZHOU L Y,HU T X,et al.A new early-maturing walnut cultivar‘Chuanzao 2’[J].Acta Horticulturae Sinica,2012,39(1):2 317-2 318.
[3]周兰英,肖千文,蒲光兰.早实核桃新品种‘蜀玲’的选育[J].安徽农业科学,2009,37(23):10 964-10 978.ZHOU L Y,XIAO Q W,PU G L.Breeding of new earlyfruiting walnut variety‘Shuling’[J].Journal of Anhui Agricultural Sciences,2009,37(23):10 964-10 978.
[4]蒲光兰,肖千文,周兰英.早实核桃新品种‘川早1号’[J].园艺学报,2011,38(10):2 025-2 026.PU G L,XIAO Q W,ZHOU L Y.A new early-fruiting walnut cultivar‘Chuanzao 1’[J].Acta Horticulturae Sinica,2011,38(10):2 025-2 026.
[5]肖千文,肖前刚,周兰英,等.早熟薄皮核桃新品种‘双早’[J].园艺学报,2013,40(1):179-180.XIAO Q W,XIAO Q G,ZHOU L Y,et al.A new early-maturing and thin shell walnut cultivar‘Shuangzao’[J].Acta Horticulturae Sinica,2013,40(1):179-180.
[6]吴佐英,白杰健,邵开茂.早实核桃新品种‘青川1号’[J].园艺学报,2013,40(12):2 531-2 532.WU Z Y,BAI J J,SHAO K M.A new precocious walnut cultivar‘Qingchuan 1’[J].Acta Horticulturae Sinica,2013,40(12):2 531-2 532.
[7]陈霞,陈少瑜,陆斌,等.分子标记及其在核桃种质资源研究中的应用[J].现代农业科技,2009,(14):347-350.CHEN X,CHEN S Y,LU B,et al.Molecular markers and its applications in germplasm research of walnut[J].Modern Agricultural Science and Technology,2009,(14):347-350.
[8]艾呈祥,秦志华,王洁,等.山东牛心柿群体遗传多样性和亲缘关系的SSR分析[J].果树学报,2013,30(4):558-562.AI C X,QIN Z H,WANG J,et al.Genetic diversity and relationship analysis of oriental persimmon(Diospyroskaki)populations in Shandong by SSR[J].Journal of Fruit Science,2013,30(4):558-562.
[9]EBRAHIMI A,FATAHI R,ZAMANI Z.Analysis of genetic diversity among some persian walnut genotypes(Juglans regia L.)using morphological traits and SSRs markers[J].Scientia Horticulturae,2011,130(1):146-151.
[10]肖志娟,翟梅枝,王振元,等.微卫星DNA在分析核桃遗传多样性上的应用[J].中南林业科技大学学报,2014,34(2):55-61.XIAO Z J,ZHAI M Z,WANG Z Y,et al.Application of microsatellite DNA on analyzing genetic diversity of Juglans regia[J].Journal of Central South University of Forestry&Technology,2014,34(2):55-61.
[11]薛华柏,杨健,王龙,等.29个梨品种SSR特征指纹数据表的构建[J].果树学报,2015,32(6):1 028-1 035.XUE H B,YANG J,WANG L,et al.Construction of SSR characteristic fingerprinting data table for 29pear cultivars[J].Journal of Fruit Science,2015,32(6):1 028-1 035.
[12]高源,刘凤之,王昆,等.苹果部分种质资源分子身份证的构建[J].中国农业科学,2015,48(19):3 887-3 898.GAO Y,LIU F Z,WANG K,et al.Establishment of molecular ID for some apple germplasm resources[J].Scientia Agricultura Sinica,2015,48(19):3 887-3 898.
[13]WANG H,WU W,PAN G,et al.Analysis of genetic diversity and relationships among 86persian walnut(Juglans regia L.)genotypes in Tibet using morphological traits and SSR markers[J].Journal of Horticultural Science&Biotechnology,2015,90(5):563-570.
[14]HAN H,WOESTE K,HU Y H,et al.Genetic diversity and population structure of common walnut(Juglans regia)in China based on EST-SSRs and the nuclear gene phenylalanine ammonia-lyase(PAL)[J].Tree Genetics&Genomes,2016,12(6):111-122.
[15]VISCHI M,CHIABA C,RARANCIUC S,et al.Genetic diversity of walnut(Juglans regia L.)in the eastern Italian alps[J].Forests,2017,8(3):81-94.
[16]陈凌娜,马庆国,张俊佩,等.核桃BES-SSR的开发及在遗传多样性分析中的应用[J].北京林业大学学报,2014,36(6):24-29.CHEN L N,MA Q G,ZHANG J P,et al.Development of BES-SSR markers in walnut and its application in genetic diversity analysis[J].Journal of Beijing Forestry University,2014,36(6):24-29.
[17]陈少瑜,宁德鲁,吴涛,等.泡核桃SSR标记开发及在遗传多样性研究中的应用[J].西北林学院学报,2017,32(3):91-96.CHEN S Y,NING D L,WU T,et al.Development of SSR markers in Juglans sigillata and its application in genetic diversity analysis[J].Journal of Northwest Forestry University,2017,32(3):91-96.
[18]史丽会,朱鹏,魏童,等.核桃品种微卫星标记的筛选与鉴别[J].果树学报,2016,33(7):783-793.SHI L H,ZHU P,WEI T,et al.SSR screening and identification of walnut(Juglans regia)cultivars[J].Journal of Fruit Science,2016,33(7):783-793.
[19]李留春.云南省核桃产业实现又好又快发展的理性思考[J].林业调查规划,2010,35(3):64-67.LI L C.Rational thinking on achieving walnut industrys a good and quick development in Yunnan Province[J].Forest Inventory and Planning,2010,35(3):64-67.
[20]韩华柏,朱益川,余凌帆,等.四川核桃生产现状与产业化发展对策[J].经济林研究,2003,21(4):138-140.HAN H B,ZHU Y C,YU L F,et al.Walnut production status and industrialization proposal for Sichuan Province[J].Economic Forest Researches,2003,21(4):138-140.
[21]CHEN L N,MA Q G,CHEN Y K,et al.Identification of major walnut cultivars grown in China based on nut phenotypes and SSR markers[J].Scientia Horticulturae,2014,(168):240-248.
[22]陈凌娜.核桃重要种质遗传分析及主栽品种指纹鉴定[D].北京:北京林业大学,2014.
[23]高源,刘凤之,曹玉芬,等.苹果属种质资源亲缘关系的SSR分析[J].果树学报,2007,24(2):129-134.GAO Y,LIU F Z,CAO Y F,et al.Analysis of genetic relationship for Malus germplasm resources by SSR markers[J].Journal of Fruit Science,2007,24(2):129-134.
[24]张瑞萍,林高星,阎振立,等.基于SSR标记的苹果砧木资源的亲缘关系分析[J].分子植物育种,2017,15(7):2 847-2 855.ZHANG R P,LIN G X,YAN Z L,et al.Genetic relationship analysis of apple rootstock resources based on SSR markers[J].Molecular Plant Breeding,2017,15(7):2 847-2 855.
[25]张明泽,姚玉仙,陈世军.黔南60份茶树种质资源遗传多样性的SSR分析[J].西北植物学报,2016,36(6):1 117-1 124.ZHANG M Z,YAO Y X,CHEN S J.Genetic diversity analysis of tea germplasm in Qiannan prefecture by SSR markers[J].Acta Boreali-Occidentalia Sinica,2016,36(6):1 117-1 124.
[26]张淑青,刘冬成,刘威生,等.普通杏品种SSR遗传多样性分析[J].园艺学报,2010,37(1):23-30.ZHANG S Q,LIU D C,LIU W S,et al.Analysis of genetic diversities in apricot cultivars(Prunus armeniaca L.)with simple sequence repeat(SSR)markers[J].Acta Horticulturae Sinica,2010,37(1):23-30.
[27]宋常美,文晓鹏,杨尔泰.贵州樱桃种质资源的ISSR分析[J].园艺学报,2011,38(8):1 531-1 538.SONG C M,WEN X P,YANG E T.Cherry germplasm from Guizhou Province analyzed by ISSR markers[J].Acta Horticulturae Sinica,2011,38(8):1 531-1 538.
[28]张志仙,何道根,朱长志,等.青花菜种质资源遗传多样性的SSR分析[J].浙江农业学报,2017,29(2):228-235.ZHANG Z X,HE D G,ZHU C Z,et al.Genetic diversity analysis of Brassica oleracea L.var.italica with SSR markers[J].Acta Agriculturae Zhejiangensis,2017,29(2):228-235.
[29]陈良华,胡庭兴,张帆,等.用AFLP技术分析四川核桃资源的遗传多样性[J].植物生态学报,2008,32(6):1 362-1 372.CHEN L H,HU T X,ZHANG F,et al.Genatic diversities of four Juglans populations revealed by AFLP in Sichuan Province,China[J].Journal of Plant Ecology,2008,32(6):1 362-1 372.
[2]肖千文,周兰英,胡庭兴,等.早实核桃新品种‘川早2号’[J].园艺学报,2012,39(11):2 317-2 318.XIAO Q W,ZHOU L Y,HU T X,et al.A new early-maturing walnut cultivar‘Chuanzao 2’[J].Acta Horticulturae Sinica,2012,39(1):2 317-2 318.
[3]周兰英,肖千文,蒲光兰.早实核桃新品种‘蜀玲’的选育[J].安徽农业科学,2009,37(23):10 964-10 978.ZHOU L Y,XIAO Q W,PU G L.Breeding of new earlyfruiting walnut variety‘Shuling’[J].Journal of Anhui Agricultural Sciences,2009,37(23):10 964-10 978.
[4]蒲光兰,肖千文,周兰英.早实核桃新品种‘川早1号’[J].园艺学报,2011,38(10):2 025-2 026.PU G L,XIAO Q W,ZHOU L Y.A new early-fruiting walnut cultivar‘Chuanzao 1’[J].Acta Horticulturae Sinica,2011,38(10):2 025-2 026.
[5]肖千文,肖前刚,周兰英,等.早熟薄皮核桃新品种‘双早’[J].园艺学报,2013,40(1):179-180.XIAO Q W,XIAO Q G,ZHOU L Y,et al.A new early-maturing and thin shell walnut cultivar‘Shuangzao’[J].Acta Horticulturae Sinica,2013,40(1):179-180.
[6]吴佐英,白杰健,邵开茂.早实核桃新品种‘青川1号’[J].园艺学报,2013,40(12):2 531-2 532.WU Z Y,BAI J J,SHAO K M.A new precocious walnut cultivar‘Qingchuan 1’[J].Acta Horticulturae Sinica,2013,40(12):2 531-2 532.
[7]陈霞,陈少瑜,陆斌,等.分子标记及其在核桃种质资源研究中的应用[J].现代农业科技,2009,(14):347-350.CHEN X,CHEN S Y,LU B,et al.Molecular markers and its applications in germplasm research of walnut[J].Modern Agricultural Science and Technology,2009,(14):347-350.
[8]艾呈祥,秦志华,王洁,等.山东牛心柿群体遗传多样性和亲缘关系的SSR分析[J].果树学报,2013,30(4):558-562.AI C X,QIN Z H,WANG J,et al.Genetic diversity and relationship analysis of oriental persimmon(Diospyroskaki)populations in Shandong by SSR[J].Journal of Fruit Science,2013,30(4):558-562.
[9]EBRAHIMI A,FATAHI R,ZAMANI Z.Analysis of genetic diversity among some persian walnut genotypes(Juglans regia L.)using morphological traits and SSRs markers[J].Scientia Horticulturae,2011,130(1):146-151.
[10]肖志娟,翟梅枝,王振元,等.微卫星DNA在分析核桃遗传多样性上的应用[J].中南林业科技大学学报,2014,34(2):55-61.XIAO Z J,ZHAI M Z,WANG Z Y,et al.Application of microsatellite DNA on analyzing genetic diversity of Juglans regia[J].Journal of Central South University of Forestry&Technology,2014,34(2):55-61.
[11]薛华柏,杨健,王龙,等.29个梨品种SSR特征指纹数据表的构建[J].果树学报,2015,32(6):1 028-1 035.XUE H B,YANG J,WANG L,et al.Construction of SSR characteristic fingerprinting data table for 29pear cultivars[J].Journal of Fruit Science,2015,32(6):1 028-1 035.
[12]高源,刘凤之,王昆,等.苹果部分种质资源分子身份证的构建[J].中国农业科学,2015,48(19):3 887-3 898.GAO Y,LIU F Z,WANG K,et al.Establishment of molecular ID for some apple germplasm resources[J].Scientia Agricultura Sinica,2015,48(19):3 887-3 898.
[13]WANG H,WU W,PAN G,et al.Analysis of genetic diversity and relationships among 86persian walnut(Juglans regia L.)genotypes in Tibet using morphological traits and SSR markers[J].Journal of Horticultural Science&Biotechnology,2015,90(5):563-570.
[14]HAN H,WOESTE K,HU Y H,et al.Genetic diversity and population structure of common walnut(Juglans regia)in China based on EST-SSRs and the nuclear gene phenylalanine ammonia-lyase(PAL)[J].Tree Genetics&Genomes,2016,12(6):111-122.
[15]VISCHI M,CHIABA C,RARANCIUC S,et al.Genetic diversity of walnut(Juglans regia L.)in the eastern Italian alps[J].Forests,2017,8(3):81-94.
[16]陈凌娜,马庆国,张俊佩,等.核桃BES-SSR的开发及在遗传多样性分析中的应用[J].北京林业大学学报,2014,36(6):24-29.CHEN L N,MA Q G,ZHANG J P,et al.Development of BES-SSR markers in walnut and its application in genetic diversity analysis[J].Journal of Beijing Forestry University,2014,36(6):24-29.
[17]陈少瑜,宁德鲁,吴涛,等.泡核桃SSR标记开发及在遗传多样性研究中的应用[J].西北林学院学报,2017,32(3):91-96.CHEN S Y,NING D L,WU T,et al.Development of SSR markers in Juglans sigillata and its application in genetic diversity analysis[J].Journal of Northwest Forestry University,2017,32(3):91-96.
[18]史丽会,朱鹏,魏童,等.核桃品种微卫星标记的筛选与鉴别[J].果树学报,2016,33(7):783-793.SHI L H,ZHU P,WEI T,et al.SSR screening and identification of walnut(Juglans regia)cultivars[J].Journal of Fruit Science,2016,33(7):783-793.
[19]李留春.云南省核桃产业实现又好又快发展的理性思考[J].林业调查规划,2010,35(3):64-67.LI L C.Rational thinking on achieving walnut industrys a good and quick development in Yunnan Province[J].Forest Inventory and Planning,2010,35(3):64-67.
[20]韩华柏,朱益川,余凌帆,等.四川核桃生产现状与产业化发展对策[J].经济林研究,2003,21(4):138-140.HAN H B,ZHU Y C,YU L F,et al.Walnut production status and industrialization proposal for Sichuan Province[J].Economic Forest Researches,2003,21(4):138-140.
[21]CHEN L N,MA Q G,CHEN Y K,et al.Identification of major walnut cultivars grown in China based on nut phenotypes and SSR markers[J].Scientia Horticulturae,2014,(168):240-248.
[22]陈凌娜.核桃重要种质遗传分析及主栽品种指纹鉴定[D].北京:北京林业大学,2014.
[23]高源,刘凤之,曹玉芬,等.苹果属种质资源亲缘关系的SSR分析[J].果树学报,2007,24(2):129-134.GAO Y,LIU F Z,CAO Y F,et al.Analysis of genetic relationship for Malus germplasm resources by SSR markers[J].Journal of Fruit Science,2007,24(2):129-134.
[24]张瑞萍,林高星,阎振立,等.基于SSR标记的苹果砧木资源的亲缘关系分析[J].分子植物育种,2017,15(7):2 847-2 855.ZHANG R P,LIN G X,YAN Z L,et al.Genetic relationship analysis of apple rootstock resources based on SSR markers[J].Molecular Plant Breeding,2017,15(7):2 847-2 855.
[25]张明泽,姚玉仙,陈世军.黔南60份茶树种质资源遗传多样性的SSR分析[J].西北植物学报,2016,36(6):1 117-1 124.ZHANG M Z,YAO Y X,CHEN S J.Genetic diversity analysis of tea germplasm in Qiannan prefecture by SSR markers[J].Acta Boreali-Occidentalia Sinica,2016,36(6):1 117-1 124.
[26]张淑青,刘冬成,刘威生,等.普通杏品种SSR遗传多样性分析[J].园艺学报,2010,37(1):23-30.ZHANG S Q,LIU D C,LIU W S,et al.Analysis of genetic diversities in apricot cultivars(Prunus armeniaca L.)with simple sequence repeat(SSR)markers[J].Acta Horticulturae Sinica,2010,37(1):23-30.
[27]宋常美,文晓鹏,杨尔泰.贵州樱桃种质资源的ISSR分析[J].园艺学报,2011,38(8):1 531-1 538.SONG C M,WEN X P,YANG E T.Cherry germplasm from Guizhou Province analyzed by ISSR markers[J].Acta Horticulturae Sinica,2011,38(8):1 531-1 538.
[28]张志仙,何道根,朱长志,等.青花菜种质资源遗传多样性的SSR分析[J].浙江农业学报,2017,29(2):228-235.ZHANG Z X,HE D G,ZHU C Z,et al.Genetic diversity analysis of Brassica oleracea L.var.italica with SSR markers[J].Acta Agriculturae Zhejiangensis,2017,29(2):228-235.
[29]陈良华,胡庭兴,张帆,等.用AFLP技术分析四川核桃资源的遗传多样性[J].植物生态学报,2008,32(6):1 362-1 372.CHEN L H,HU T X,ZHANG F,et al.Genatic diversities of four Juglans populations revealed by AFLP in Sichuan Province,China[J].Journal of Plant Ecology,2008,32(6):1 362-1 372.