基于SSR分子标记的桉树父本鉴定技术研究
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摘要
利用筛选出的11对SSR引物进行SSR分子标记分析,通过比对子代与母本相同引物对应标记位点的等位基因配置,鉴别自由授粉过程中可能由母本自交产生的个体,并借助Cervus软件应用最大似然值法和遗传排除法,从154个基因型候选父本中鉴定出D2、D3、D4、D7、D11、D19和D20等7个半同胞子代的遗传父本分别为同一试验林内定植的尾巨桉DH18-04、DH18-20、DH32-29和DH241-2。研究对于今后进一步深入探究桉树自交不亲和与近交衰退机制,准确进行子代父本鉴定和杂交亲本重建,以及品种选育等工作具有重要指导意义。
A technique for male parent identification using SSR markers in Eucalyptus half-sib progenies was developed in the study reported here. Eleven selected SSR loci with high polymorphism were used to identify the selfed individuals in an open pollinated population, based on the comparison of marker loci with the same primers between progenies and female parents. According to maximum likelihood scores obtained from analyses by Cervus software, E.urophylla × E. grandis clones DH18-04, DH18-20, DH32-29 and DH241-2 were identified as the male parents of the outcrossed progenies D2, D3, D4, D7, D11 and D20, respectively.This research has important significance for guiding further examination of self-incompatibility, paternal identification and hybrid parental reconstruction in these eucalypt taxa.
A technique for male parent identification using SSR markers in Eucalyptus half-sib progenies was developed in the study reported here. Eleven selected SSR loci with high polymorphism were used to identify the selfed individuals in an open pollinated population, based on the comparison of marker loci with the same primers between progenies and female parents. According to maximum likelihood scores obtained from analyses by Cervus software, E.urophylla × E. grandis clones DH18-04, DH18-20, DH32-29 and DH241-2 were identified as the male parents of the outcrossed progenies D2, D3, D4, D7, D11 and D20, respectively.This research has important significance for guiding further examination of self-incompatibility, paternal identification and hybrid parental reconstruction in these eucalypt taxa.
引文
[1]RICHARD P,李海佳,李宏伟,等.广西桉树人工杂交育种研究[J].广西林业科学,2006,35(4):238-242.
[2]陈小勇.自然植物种群的亲本分析及其在生态学研究中的应用[J].生态学杂志,1999,18(2):30-35.
[3]POLLEGIONI P,WOESTE K,MUGNOZZA G S,et al.Retrospective identification of hybridogenic walnut plants by SSR fingerprinting and parentage analysis[J].Molecular Breeding,2009,24(4):321-335.
[4]MEAGHER T R.Analysis of paternity within a natural population of Chamaelirium luteum.1.Identification of most-likely male parents[J].The American Naturalist,1986,128(2):199-215.
[5]MEAGHER T R,THOMPSON E.The relationship between single parent and parent pair genetic likelihoods in genealogy reconstruction[J].Theoretical Population Biology,1986,29(1):87-106.
[6]MARSHALL T C,SLATE J,KRUUK L,et al.Statistical confidence for likelihood-based paternity inference in natural populations[J].Molecular Ecology,1998,7(5):639-655.
[7]JONES A G,SMALL C M,PACZOLT K A,et al.Apractical guide to methods of parentage analysis[J].Molecular Ecology Resources,2010,10(1):6-30.
[8]ROBLEDO-A J J,GIL L.Patterns of pollen dispersal in a small population of Pinus sylvestris L.revealed by totalexclusion paternity analysis[J].Heredity,2005,94(1):13-22.
[9]REIZ C,BRETO M P,ASINS M J.A quick methodology to identify sexual seedlings in citrus breeding programs using SSR marker[J].Euphytica,2000,112(1):89-94.
[10]GRATTAPAGLIA D,RIBEIRO V J,REZENDE G D S P.Retrospective selection of elite parent trees using paternity testing with microsatellite markers:an alternative short term breeding tactic for Eucalyptus[J].Theoretical Applied Genetics,2004,109(1):192-199.
[11]HAN Z,GAO P,GENG X,et al.Identification of the male parent of superior half-sib Populus tomentosa individuals based on SSR markers[J].Molecular Breeding,2017,37(12):155-166.
[12]CHAIX G,GERBER S,RAZAFIMAHARO V,et al.Gene flow estimation with microsatellites in a Malagasy seed orchard of Eucalyptus grandis[J].Theoretical Applied Genetics,2003,107(4):705-712.
[13]PATTERSOM B,GORE P,POTTS B M,et al.Advances in pollination techniques for large-scale seed production in Eucalyptus globules[J].Australian Journal of Botany,2004,52(6):781-788.
[14]GRATTAPAGLIA D.Molecular breeding of Eucalyptus Molecular biology of woody plants[M].Netherlands:Springer,2000.
[15]HULCE D,LI X,SNYDER-L T.Gene Marker?genotyping software:Tools to increase the statistical power of DNA fragment analysis[J].Journal of Biomolecular Techniques:JBT,2011,22(Suppl):35-36.
[16]ZHOU C,HE X,LI F,et al.Development of 240 novel EST-SSRs in Eucalyptus L’Herit[J].Molecular Breeding,2014,33(1):221-225.
[17]YEH F,YANG R,BOYLE T.POPGENE Version 1.32Microsoft Windows-based freeware for populations genetic analysis[M].Edmonton:University of Alberta,1999.
[18]ISAGI Y,KANAZASHI T,SUZUKI W,et al.Highly variable pollination patterns in Magnolia obovata revealed by microsatellite paternity analysis[J].International Journal of Plant Sciences,2004,165(6):1047-1053.
[19]VAN E A L,WEABER R L,DRAKE D J,et al.DNA-based paternity analysis and genetic evaluation in a large,commercial cattle ranch setting[J].Journal of Animal Science,2007,85(12):3159-3169.
[20]GRIFFIN A R,HAND F C.Post-anthesis development of flowers of Eucalyptus regnans F.Muell and the timing of artificial pollination[J].Australian Forest Research,1979,9:9-15.
[21]ELDRIDGE K G,DAVIDSON J,HARDWOOD C,et al.Eucalypt domestication and breeding[M].Oxford:Clarendon Press,1993.
[22]POTTS B M,GORE P L.Reproductive biology and controlled pollination of Eucalyptus-a review[M].Tasmania:School of Plant Science UTAS,1995.
[23]LARCOMBE M J,COSTA J,TILYARD P,et al.On the persistence of reproductive barriers in Eucalyptus:The bridging of mechanical barriers to zygote formation by F1hybrids is counteracted by intrinsic post-zygotic incompatibilities[J].Annals of Botany,2016,118(3):431-444.
[24]POUND L M,WALLWORK M A B,POTTS B M,et al..Early ovule development following self-and cross-pollinations in Eucalyptus globulus Labill.ssp.globules[J].Annals of Botany,2002,89(5):613-620.
[25]HARDNER C M,POTTS B M.Postdispersal selection following mixed mating in Eucalyptus regnans[J].Evolution,1997,51(1):103-111.
[26]COSTA E S J,HARDNER C,TILYARD P,et al.The effects of age and environment on the expression of inbreeding depression in Eucalyptus globulus[J].Heredity,2011,107(1):50-60.
[27]WILLIAMS D R,POTTS B M,BLACK P G.Testing single visit pollination procedures for Eucalyptus globulus and E.nitens[J].Australian Forestry,1999,62(4):346-352.
[28]HARBARD J L,GRIFFIN A R,ESPEJO J.Mass controlled pollination of Eucalyptus globulus:a practical reality[J].Canadian Journal Forestry Research,1999,29(10):1457-1463.
[29]HORSLEY T N,JOHNSON S D,MYBURG A A.Comparison of different control-pollination techniques for small-flowered eucalypts[J].New Forests,2010,39(1):75-88.
[30]HARDNER C M,Potts B M.Inbreeding depression and changes in variation after selfing in Eucalyptus globulus ssp.globules[J].Silvae Genetica,1995,44:46-54.
[31]DOUGHTY R W.The Eucalyptus.A natural and commercial history of the gum tree[M].Baltimore:John Hopkins Univ Press,2000.
[32]WILCOX M D.A catalogue of the eucalypts[M].Auckland:Groome Poyry,1997.
[33]DONG C B,SUO Y J,KANG X Y.Assessment of the genetic composition of triploid hybrid Populus using SSRmarkers with low recombination frequencies[J].Canadian Journal Forestry Research,2014,44(7):692-699.
[34]BARBOUR R C,POTTS B M,VAILLANCOUNT R E.Gene flow between introduced and native Eucalyptus species:crossability of native Tasmanian species with exotic E.nitens[J].Australian Journal of Botany,2005,53(5):465-477.
[35]BARBOUR R C,POTTS B M,Vaillancourt RE.Gene flow between introduced and native Eucalyptus species:Early-age selection limits invasive capacity of exotic E.ovate×nitens F1 hybrids[J].Forest Ecology and Management,2006,228:206-214.
[36]BARBOUR R C,POTTS B M,VAILLANCOUNT R E.Gene flow between introduced and native Eucalyptus species:Morphological analysis of tri-species and backcross hybrids involving E.nitens[J].Silvae Genetica,2007,56(3/4):127-133.
[2]陈小勇.自然植物种群的亲本分析及其在生态学研究中的应用[J].生态学杂志,1999,18(2):30-35.
[3]POLLEGIONI P,WOESTE K,MUGNOZZA G S,et al.Retrospective identification of hybridogenic walnut plants by SSR fingerprinting and parentage analysis[J].Molecular Breeding,2009,24(4):321-335.
[4]MEAGHER T R.Analysis of paternity within a natural population of Chamaelirium luteum.1.Identification of most-likely male parents[J].The American Naturalist,1986,128(2):199-215.
[5]MEAGHER T R,THOMPSON E.The relationship between single parent and parent pair genetic likelihoods in genealogy reconstruction[J].Theoretical Population Biology,1986,29(1):87-106.
[6]MARSHALL T C,SLATE J,KRUUK L,et al.Statistical confidence for likelihood-based paternity inference in natural populations[J].Molecular Ecology,1998,7(5):639-655.
[7]JONES A G,SMALL C M,PACZOLT K A,et al.Apractical guide to methods of parentage analysis[J].Molecular Ecology Resources,2010,10(1):6-30.
[8]ROBLEDO-A J J,GIL L.Patterns of pollen dispersal in a small population of Pinus sylvestris L.revealed by totalexclusion paternity analysis[J].Heredity,2005,94(1):13-22.
[9]REIZ C,BRETO M P,ASINS M J.A quick methodology to identify sexual seedlings in citrus breeding programs using SSR marker[J].Euphytica,2000,112(1):89-94.
[10]GRATTAPAGLIA D,RIBEIRO V J,REZENDE G D S P.Retrospective selection of elite parent trees using paternity testing with microsatellite markers:an alternative short term breeding tactic for Eucalyptus[J].Theoretical Applied Genetics,2004,109(1):192-199.
[11]HAN Z,GAO P,GENG X,et al.Identification of the male parent of superior half-sib Populus tomentosa individuals based on SSR markers[J].Molecular Breeding,2017,37(12):155-166.
[12]CHAIX G,GERBER S,RAZAFIMAHARO V,et al.Gene flow estimation with microsatellites in a Malagasy seed orchard of Eucalyptus grandis[J].Theoretical Applied Genetics,2003,107(4):705-712.
[13]PATTERSOM B,GORE P,POTTS B M,et al.Advances in pollination techniques for large-scale seed production in Eucalyptus globules[J].Australian Journal of Botany,2004,52(6):781-788.
[14]GRATTAPAGLIA D.Molecular breeding of Eucalyptus Molecular biology of woody plants[M].Netherlands:Springer,2000.
[15]HULCE D,LI X,SNYDER-L T.Gene Marker?genotyping software:Tools to increase the statistical power of DNA fragment analysis[J].Journal of Biomolecular Techniques:JBT,2011,22(Suppl):35-36.
[16]ZHOU C,HE X,LI F,et al.Development of 240 novel EST-SSRs in Eucalyptus L’Herit[J].Molecular Breeding,2014,33(1):221-225.
[17]YEH F,YANG R,BOYLE T.POPGENE Version 1.32Microsoft Windows-based freeware for populations genetic analysis[M].Edmonton:University of Alberta,1999.
[18]ISAGI Y,KANAZASHI T,SUZUKI W,et al.Highly variable pollination patterns in Magnolia obovata revealed by microsatellite paternity analysis[J].International Journal of Plant Sciences,2004,165(6):1047-1053.
[19]VAN E A L,WEABER R L,DRAKE D J,et al.DNA-based paternity analysis and genetic evaluation in a large,commercial cattle ranch setting[J].Journal of Animal Science,2007,85(12):3159-3169.
[20]GRIFFIN A R,HAND F C.Post-anthesis development of flowers of Eucalyptus regnans F.Muell and the timing of artificial pollination[J].Australian Forest Research,1979,9:9-15.
[21]ELDRIDGE K G,DAVIDSON J,HARDWOOD C,et al.Eucalypt domestication and breeding[M].Oxford:Clarendon Press,1993.
[22]POTTS B M,GORE P L.Reproductive biology and controlled pollination of Eucalyptus-a review[M].Tasmania:School of Plant Science UTAS,1995.
[23]LARCOMBE M J,COSTA J,TILYARD P,et al.On the persistence of reproductive barriers in Eucalyptus:The bridging of mechanical barriers to zygote formation by F1hybrids is counteracted by intrinsic post-zygotic incompatibilities[J].Annals of Botany,2016,118(3):431-444.
[24]POUND L M,WALLWORK M A B,POTTS B M,et al..Early ovule development following self-and cross-pollinations in Eucalyptus globulus Labill.ssp.globules[J].Annals of Botany,2002,89(5):613-620.
[25]HARDNER C M,POTTS B M.Postdispersal selection following mixed mating in Eucalyptus regnans[J].Evolution,1997,51(1):103-111.
[26]COSTA E S J,HARDNER C,TILYARD P,et al.The effects of age and environment on the expression of inbreeding depression in Eucalyptus globulus[J].Heredity,2011,107(1):50-60.
[27]WILLIAMS D R,POTTS B M,BLACK P G.Testing single visit pollination procedures for Eucalyptus globulus and E.nitens[J].Australian Forestry,1999,62(4):346-352.
[28]HARBARD J L,GRIFFIN A R,ESPEJO J.Mass controlled pollination of Eucalyptus globulus:a practical reality[J].Canadian Journal Forestry Research,1999,29(10):1457-1463.
[29]HORSLEY T N,JOHNSON S D,MYBURG A A.Comparison of different control-pollination techniques for small-flowered eucalypts[J].New Forests,2010,39(1):75-88.
[30]HARDNER C M,Potts B M.Inbreeding depression and changes in variation after selfing in Eucalyptus globulus ssp.globules[J].Silvae Genetica,1995,44:46-54.
[31]DOUGHTY R W.The Eucalyptus.A natural and commercial history of the gum tree[M].Baltimore:John Hopkins Univ Press,2000.
[32]WILCOX M D.A catalogue of the eucalypts[M].Auckland:Groome Poyry,1997.
[33]DONG C B,SUO Y J,KANG X Y.Assessment of the genetic composition of triploid hybrid Populus using SSRmarkers with low recombination frequencies[J].Canadian Journal Forestry Research,2014,44(7):692-699.
[34]BARBOUR R C,POTTS B M,VAILLANCOUNT R E.Gene flow between introduced and native Eucalyptus species:crossability of native Tasmanian species with exotic E.nitens[J].Australian Journal of Botany,2005,53(5):465-477.
[35]BARBOUR R C,POTTS B M,Vaillancourt RE.Gene flow between introduced and native Eucalyptus species:Early-age selection limits invasive capacity of exotic E.ovate×nitens F1 hybrids[J].Forest Ecology and Management,2006,228:206-214.
[36]BARBOUR R C,POTTS B M,VAILLANCOUNT R E.Gene flow between introduced and native Eucalyptus species:Morphological analysis of tri-species and backcross hybrids involving E.nitens[J].Silvae Genetica,2007,56(3/4):127-133.