湿地松×洪都拉斯加勒比松遗传多样性的ISSR分析（英文）
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摘要
杂交育种是产生遗传变异、表型变异及选择新变异的重要方法。然而系统发育不清晰,选择较近的亲缘关系亲本用于杂交子代往往表现出较低的遗传多样性。为探究湿地松、洪都拉斯加勒比松种间杂交后代遗传多样性水平,对8个湿地松×洪都拉斯加勒比松家系进行ISSR分析。利用10条引物共产生60个表达清晰可用于分析的标记,其中48个标记表现为多态性,占总标记数的80%;湿加松各个家系多态位点百分率在5%～23.33%之间;各个家系基因多样性指数在0.015 2～0.087 2之间,Shannon指数的范围在0.021 6～0.129 4之间(家系水平为0.293 4)。8个家系间的基因分化系数Gst为0.743 5,即总的遗传变异中有74.35%的变异存在于家系间,家系内的遗传变异占总遗传变异的25.65%。采用UPGMA法对湿加松的8个家系进行了聚类分析,确定了各个家系之间的遗传亲缘关系。8个家系间的基因流N_m为0.172 5,表明基因流处于较低水平。
Cross-breeding is an important method for generating genetic and phenotypic variation for selecting new varieties. However,because of an uncertainty of phylogenetic relationships,the parents selected for crosses may have a close genetic relationship resulting in hybrid progeny that shows low genetic diversity. Analysis of inter-family genetic diversity was undertaken among eight Pinus elliottii × P. caribaea var. hondurensis full-sib families using inter-sequence simple repeat(ISSR) markers. A total of 480 individuals were analyzed using 10 ISSR primers. Nei's unbiased gene diversity in the families ranged from 0.015 2 to 0.087 2. Shannon genetic diversity index values ranged from 0.021 6 to 0.129 4. Only a small proportion(25. 65%) of genetic variation resided within families,whereas the majority of genetic variation(74.35%) accounted for the inter-family genetic differentiation index of Gst= 0.743 5. On the basis of estimated genetic distance and UPGMA clustering analysis,the genetic differentiation among the eight families was indicated to be relatively high with low gene flow(N_m= 0.172 5). The low inter-family gene flow may be related to the high genetic heterozygosity of slash pine and Caribbean pine. These findings are expected to provide a foundation for genetic breeding of Pinus elliottii × P. caribaea var. hondurensis hybrids.
Cross-breeding is an important method for generating genetic and phenotypic variation for selecting new varieties. However,because of an uncertainty of phylogenetic relationships,the parents selected for crosses may have a close genetic relationship resulting in hybrid progeny that shows low genetic diversity. Analysis of inter-family genetic diversity was undertaken among eight Pinus elliottii × P. caribaea var. hondurensis full-sib families using inter-sequence simple repeat(ISSR) markers. A total of 480 individuals were analyzed using 10 ISSR primers. Nei's unbiased gene diversity in the families ranged from 0.015 2 to 0.087 2. Shannon genetic diversity index values ranged from 0.021 6 to 0.129 4. Only a small proportion(25. 65%) of genetic variation resided within families,whereas the majority of genetic variation(74.35%) accounted for the inter-family genetic differentiation index of Gst= 0.743 5. On the basis of estimated genetic distance and UPGMA clustering analysis,the genetic differentiation among the eight families was indicated to be relatively high with low gene flow(N_m= 0.172 5). The low inter-family gene flow may be related to the high genetic heterozygosity of slash pine and Caribbean pine. These findings are expected to provide a foundation for genetic breeding of Pinus elliottii × P. caribaea var. hondurensis hybrids.
引文
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DVORAK WS,POTTER KM,HIPKINS VD,et al,2009.Genetic diversity and gene exchange in Pinus oocarpa,a Mesoamerican pine with resistance to the pitch canker fungus(Fusarium circinatum)[J].Inter J Plant Sci,170(5):609-626.
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HAMELIN RC,BEAULICU J,PLOURDE A,1995.Genetic diversity in populations of cronartium ribicola in plantations and natural stands of Pinus strobes[J].Theor Appl Genet,91(8):1214-1221.
LERCETEAU E,SZMIDT AE,1999.Properties of AFLPmarkers in inheritance and genetic diversity studies of Pinus sylvestris L.[J].Heredity,82:252-260.
LI YL,ZHAO FC,ZHANG YZ,et al,2010.Rapid DNAextraction method suitable for SSR analysis in slash pine and caribbean pine[J].Biotechnol Bull,1:83-86.[李义良,赵奋成,张应中,等,2010.适用于微卫星标记的湿地松、加勒比松DNA快速提取法[J].生物技术通报,1:83-86.]
MARIETTE S,CHAGND,LZIER,et al,2001.Genetic diversity within and among Pinus pinaster populations:comparison between AFLP and microsatellite markers[J].Heredity,86:469-479.
NIKLES DG,1995.Hybirds of the Slash-Caribbean-Central American pine complex:characteristics,bases of superiority and potential utility in South China and elsewhere[M]//SHEN XH.Forest Tree Improvement in the Asia-Pacific Region.China Forestry Publishing House:168-186.
REDDY MP,SARLA N,SIDDIQ EA,2002.Inter simple sequence repeat(ISSR)polymorphism and its application in plant breeding[J].Euphytica,128:9-17.
SHAO D,PEI Y,ZHANG HQ,2007.Cp SSR Analysis of variation of genetic diversity in temporal dimension of natural population of Pinus koraiensis in liangshui national nature reserve[J].Bull Bot Res,93(4):969-980.
SHUI J,HUANG SW,CHEN BQ,2005.RAPD analysis on the genetic diversity of original provenances and internal populations of Pinus taeda[J].J S Chin Agric Univ,26(3):74-76,81.[税珺,黄少伟,陈炳铨,2005.火炬松原生种源和引种群体RAPD遗传多样性[J].华南农业大学学报,26(3):74-76,81.]
SZMIDT AE,WANG XR,LU MZ,1996.Empirical assessment of allozyme and RAPD variation in Pinus sylvestris(L.)using haploid tissue analysis[J].Heredity,76:412-420.
TANG JJ,FAN YR,ZHU MY,2003.Analysis of the genetic diversity of Pinus taiwanensis populations[J].J Zhejiang For Coll,20(1):23-26.[唐娟娟,范义荣,朱睦元,2003.黄山松群体遗传多样性分析[J].浙江林学院学报,20(1):3-26.]
WRIGHT S,1931.Evolution in Mendelian population[J].Genetics,16:97-159.
YANG CP,WEI L,JIANG J,et al,2005.Analysis of genetic diversity for nineteen populations of Pinus sibirica du tour with technique of ISSR[J].J NE For Univ,33(1):1-3.[杨传平,魏利,姜静,等,2005.应用ISSR-PCR对西伯利亚红松19个种源的遗传多样性分析[J].东北林业大学学报,33(1):1-3.]
ZHANG HG,ZHANG L,ZHU H Y,et al,2011.ISSR idetification technology of hybrid larch progenies and their parents[J].J NE For Univ,39(7):1-4.[张含国,张磊朱,航勇,等,2011.落叶松杂种与亲本ISSR鉴别技术[J].东北林业大学学报,39(7):1-4]
ZHANG W,GONG J,JI KS,2008.Genetic diversity for seedling orchard of masson’s pine[J].Mol Plant Breed,6:717-723.
ZHAO FC,LI XZ,ZHANG YZ,et al,2009.Research results and prospects of crossing breeding of slash pine×caribbean pine[J].Chin Sci Technol Achiev,10(8):21-23.[赵奋成,李宪政,张应中,等,2009.湿地松×加勒比松杂交育种研究成果与展望[J].中国科技成果,10(8):21-23.]
ZHOU FM,FAN JF,HOU WW,2008.Genetic diversity in Pinus tabulaeformis natural population in Shaanxi by RAPDmarkers[J].J NE For Univ,36(12):1-3.[周飞梅,樊军锋,侯万伟,2008.陕西地区油松天然群体遗传结构的RAPD分析[J].东北林业大学学报,36(12):1-3.]
ZHU BF,CHEN DX,CHEN YL,et al,2007.Study on the genetic diversity of seed orchard of Pinus massoniana in Guangdong Province[J].J Fujian For Sci Technol,34(3):1-5,22.[朱必凤,陈德学,陈虞禄,等,2007.广东韶关马尾松种子园遗传多样性分析[J].福建林业科技,34(3):1-5,22.]
BRAHM B,MIKLSSY I,KOVCS E,et al,2010.Genetic analysis of Pinus sylvestris L.and Pinus sylvestris forma turfosa L.using RAPD markers[J].Not Sci Biol,2(1):129-132.
DVORAK WS,POTTER KM,HIPKINS VD,et al,2009.Genetic diversity and gene exchange in Pinus oocarpa,a Mesoamerican pine with resistance to the pitch canker fungus(Fusarium circinatum)[J].Inter J Plant Sci,170(5):609-626.
FENG FJ,CHEN MM,ZHANG DD,et al,2009.Application of SRAP in the genetic diversity of Pinus koraiensis of different provenances[J].Afr J Biotechnol,8(6):1000-1008.
FENG FJ,ZHANG DD,HAN SJ,2007.Genetic diversity of superior clones from Pinus koraiensis seed orchard[J].J NEFor Univ,35(9):9-11.[冯富娟,张冬东,韩士杰,2007.红松种子园优良无性系的遗传多样性[J].东北林业大学学报,35(9):9-11.]
GODWIN ID,AITKEN EAB and SMITH LW,1997.Application of inter simple sequence repeat(ISSR)markers to plant genetics[J].Electrophoresis,18:1524-1528.
HAMELIN RC,BEAULICU J,PLOURDE A,1995.Genetic diversity in populations of cronartium ribicola in plantations and natural stands of Pinus strobes[J].Theor Appl Genet,91(8):1214-1221.
LERCETEAU E,SZMIDT AE,1999.Properties of AFLPmarkers in inheritance and genetic diversity studies of Pinus sylvestris L.[J].Heredity,82:252-260.
LI YL,ZHAO FC,ZHANG YZ,et al,2010.Rapid DNAextraction method suitable for SSR analysis in slash pine and caribbean pine[J].Biotechnol Bull,1:83-86.[李义良,赵奋成,张应中,等,2010.适用于微卫星标记的湿地松、加勒比松DNA快速提取法[J].生物技术通报,1:83-86.]
MARIETTE S,CHAGND,LZIER,et al,2001.Genetic diversity within and among Pinus pinaster populations:comparison between AFLP and microsatellite markers[J].Heredity,86:469-479.
NIKLES DG,1995.Hybirds of the Slash-Caribbean-Central American pine complex:characteristics,bases of superiority and potential utility in South China and elsewhere[M]//SHEN XH.Forest Tree Improvement in the Asia-Pacific Region.China Forestry Publishing House:168-186.
REDDY MP,SARLA N,SIDDIQ EA,2002.Inter simple sequence repeat(ISSR)polymorphism and its application in plant breeding[J].Euphytica,128:9-17.
SHAO D,PEI Y,ZHANG HQ,2007.Cp SSR Analysis of variation of genetic diversity in temporal dimension of natural population of Pinus koraiensis in liangshui national nature reserve[J].Bull Bot Res,93(4):969-980.
SHUI J,HUANG SW,CHEN BQ,2005.RAPD analysis on the genetic diversity of original provenances and internal populations of Pinus taeda[J].J S Chin Agric Univ,26(3):74-76,81.[税珺,黄少伟,陈炳铨,2005.火炬松原生种源和引种群体RAPD遗传多样性[J].华南农业大学学报,26(3):74-76,81.]
SZMIDT AE,WANG XR,LU MZ,1996.Empirical assessment of allozyme and RAPD variation in Pinus sylvestris(L.)using haploid tissue analysis[J].Heredity,76:412-420.
TANG JJ,FAN YR,ZHU MY,2003.Analysis of the genetic diversity of Pinus taiwanensis populations[J].J Zhejiang For Coll,20(1):23-26.[唐娟娟,范义荣,朱睦元,2003.黄山松群体遗传多样性分析[J].浙江林学院学报,20(1):3-26.]
WRIGHT S,1931.Evolution in Mendelian population[J].Genetics,16:97-159.
YANG CP,WEI L,JIANG J,et al,2005.Analysis of genetic diversity for nineteen populations of Pinus sibirica du tour with technique of ISSR[J].J NE For Univ,33(1):1-3.[杨传平,魏利,姜静,等,2005.应用ISSR-PCR对西伯利亚红松19个种源的遗传多样性分析[J].东北林业大学学报,33(1):1-3.]
ZHANG HG,ZHANG L,ZHU H Y,et al,2011.ISSR idetification technology of hybrid larch progenies and their parents[J].J NE For Univ,39(7):1-4.[张含国,张磊朱,航勇,等,2011.落叶松杂种与亲本ISSR鉴别技术[J].东北林业大学学报,39(7):1-4]
ZHANG W,GONG J,JI KS,2008.Genetic diversity for seedling orchard of masson’s pine[J].Mol Plant Breed,6:717-723.
ZHAO FC,LI XZ,ZHANG YZ,et al,2009.Research results and prospects of crossing breeding of slash pine×caribbean pine[J].Chin Sci Technol Achiev,10(8):21-23.[赵奋成,李宪政,张应中,等,2009.湿地松×加勒比松杂交育种研究成果与展望[J].中国科技成果,10(8):21-23.]
ZHOU FM,FAN JF,HOU WW,2008.Genetic diversity in Pinus tabulaeformis natural population in Shaanxi by RAPDmarkers[J].J NE For Univ,36(12):1-3.[周飞梅,樊军锋,侯万伟,2008.陕西地区油松天然群体遗传结构的RAPD分析[J].东北林业大学学报,36(12):1-3.]
ZHU BF,CHEN DX,CHEN YL,et al,2007.Study on the genetic diversity of seed orchard of Pinus massoniana in Guangdong Province[J].J Fujian For Sci Technol,34(3):1-5,22.[朱必凤,陈德学,陈虞禄,等,2007.广东韶关马尾松种子园遗传多样性分析[J].福建林业科技,34(3):1-5,22.]