基于ISSR标记的咖啡资源遗传多样性分析
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摘要
采用ISSR分子标记对87份咖啡资源进行遗传多样性和亲缘关系分析。结果表明,19条ISSR引物扩增出140条带,其中多态性带107条,多态性位点为76.4%。运用UPGMA方法构建了聚类图,在遗传相似系数0.625水平下,87份资源被分为3大类。第Ⅰ类群包括了3份大粒种(CoffealibericaBullexHiern);第Ⅱ类群为中粒种咖啡(C.canephora Pierre);第Ⅲ类群包括了全部小粒种资源(C. arabica Linne),共83份。咖啡种质的遗传关系种间容易划分,在种的分类水平上存在遗传关系多样性,而小粒种咖啡种内遗传多样性较窄。该研究结果将为咖啡种质鉴定、分类及分子育种提供重要科学依据。表明用ISSR分子标记进行咖啡资源遗传多样性的分析是可行的。
To assess the genetic diversity and the relationship of coffee germplasm accessions, 87 resources were analyzed by ISSR molecular marker technology. The results demonstrated that 140 bands were amjplified by 19 primers,among which 107 bands were polymorphic, accounting for 76.4%. Based on a clustering graph using UPGMA, 87 resources could be divided into three groups at the level of 0.625. Three accessions in C. liberica were in the first group.One accessions in C. canephora was in the second group. A total of 83 accessions in C. arabica were in the third group.There were large genetic differences among species(C. arabica, C. canephora, and C. liberica), and the small genetic differences within Coffea arabica. These results would provide an important scientific basis for pineapple germplasm identification, classification and molecular breeding of coffee. The ISSR markers was an available method to study the genetic diversity of coffee germplasm resources.
To assess the genetic diversity and the relationship of coffee germplasm accessions, 87 resources were analyzed by ISSR molecular marker technology. The results demonstrated that 140 bands were amjplified by 19 primers,among which 107 bands were polymorphic, accounting for 76.4%. Based on a clustering graph using UPGMA, 87 resources could be divided into three groups at the level of 0.625. Three accessions in C. liberica were in the first group.One accessions in C. canephora was in the second group. A total of 83 accessions in C. arabica were in the third group.There were large genetic differences among species(C. arabica, C. canephora, and C. liberica), and the small genetic differences within Coffea arabica. These results would provide an important scientific basis for pineapple germplasm identification, classification and molecular breeding of coffee. The ISSR markers was an available method to study the genetic diversity of coffee germplasm resources.
引文
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[11]李冬梅.饲草型小黑麦的遗传图谱构建及草产量和抗锈病相关基因的QTL定位[D].兰州:甘肃农业大学,2016.
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[13]Aga E,Bekele E,Bryngelsson T.Inter-simple sequence repeat(ISSR)variation in forest coffee trees(Coffea arabica L.)populations from Ethiopia[J].Genetica,2005,124(2-3):213-221.
[14]Masumbuko L I,Bryngelsson T.Inter simple sequence repeat(ISSR)analysis of diploid coffee species and cultivated Coffea arabica L.from Tanzania[J].Genetic Resources and Crop Evolution,2006,53(2):357-366.
[15]Kumar S A,Sudisha J,Sreenath H L.Genetic relation of Coffea and Indian Psilanthus species as revealed through RAPD and ISSR markers[J].International Journal of Integrative Biology,2008,3(3):150-158.
[16]Tshilenge P,Nkongolo K K,Mehes M,et al.Genetic variation in Coffea canephora L.(Var.Robusta)accessions from the founder gene pool evaluated with ISSR and RAPD[J].African Journal of Biotechnology,2009,8(3):380-390.
[17]Kassahun T,Kim G,Endashaw B,et al.ISSR fingerprinting of Coffea arabica throughout Ethiopia reveals high variability in wild populations and distinguishes them from landraces[J].Plant Systematics and Evolution,2014,300(5):881-897.
[18]周华,张洪波,夏红云,等.咖啡种质资源多样性研究[J].中国热带农业,2015,5:48-52.
[19]黄丽芳,董云萍,王晓阳,等.利用RAPD标记分析咖啡种质资源的遗传多样性[J].热带作物学报,2014,35(12):2313-2319.
[20]黄丽芳,董云萍,王晓阳,等.云南咖啡资源遗传多样性的RAPD分析[J].中国热带农业,2017,5:48-52.
[21]黄丽芳,董云萍,王晓阳,等.咖啡种质资源DNA指纹图谱的构建[J].热带农业科学,2016,(36),12:48-52.
[22]黄丽芳,闫林,董云萍,等.咖啡叶片DNA提取方法的比较研究[J].热带农业科学,2011,(31),12:42-45.
[23]闫林,黄丽芳,谭乐和,等.咖啡ISSR与RAPD-PCR反应体系优化[J].热带作物学报,2012,33(5):854-859.
[24]王野,陈磊,白云,等.云杉矮槲寄生遗传多样性的ISSR分析[J].西北植物学报,2017,37(11):2153-2162.
[25]时圣明,潘明佳,王洁,等.分子鉴定技术在中药中的应用[J].中草药,2016,47(17):3121-3126.
[26]马克平.试论生物多样性的概念[J].生物多样性,1993,1(1):20-22.
[27]郑勇民,张旭业,徐秉良,等.美洲南瓜种质资源遗传多样性的ISSR分析[J].甘肃农业大学学报,2017,6:76-82.
[28]Masumbuko L I,Bryngelsson T,Mneney E E.Genetic diversity in Tanzanian Arabica coffee using random amplified polymorphic DNA(RAPD)markers[J].Hereditas,2010,139(1):56-63.
[29]Rotondi A,Magli M,Ricciolini C,et al.Morphological andmolecular analyses for the characterization of a group of Italyolive cuhivars[J].Euphytica,2003,132(2):129-137.
[30]陈海云,宁德鲁,李勇杰,等.59个油橄榄种质的ISSR分子鉴定[J].东北林业大学学报,2013,41(3):13-17.
[31]席春奕,唐茜,吴永胜,等.30份四川茶树种质资源遗传多样性与亲缘关系的SRAP分析[J].贵州农业科学,2013,41(2):6-9.
[2]Ceja-Navarro J A,Vega F E,Karaoz U,et al.Gut microbiota mediate caffeine detoxification in the primary insect pest of coffee[J].Nature Communication,2015,6(7618):1-9.
[3]Stoffelen P,Noirot M,Couturon E,et al.Coffea anthonyi;a new self-compatible Central African coffee species,closely related to an ancestor of Coffea arabica[J].Taxon,2009,58(1):133-140.
[4]Nowak M D,Davis A P,Anthony F,et al.Expression and trans-specific polymorphism of self-incompatibility rnases in Coffea(Rubiaceae)[J].PLoS One,2011,6(6):1-11.
[5]Anthony F,Combes C,Astorga C,et al.The origin of cultivated Coffea arabica L.varieties revealed by AFLP and SSRmarkers[J].Theoretical&Applied Genetics.2002,104(5):894-900.
[6]Silvestrini M,Junqueira M G,Favarin A C,et al.Genetic diversity and structure of Ethiopian,Yemen and Brazilian Coffea arabica L.accessions using microsatellites markers[J].Genetic Resources and Crop Evolution,2007,54(1):1367-1379.
[7]Cenci A,Combes M C,Lashermes P.Genome evolution in diploid and tetraploid coffea species as revealed by comparative analysis of orthologous genome segments[J].Plant Molecular Biology,2012,78(1):135-145.
[8]Culley T M,Wolfe A D.Population genetic structuer of the cletstogamous plant species Viola pubescens Aiton(Vilaceae),as indicated by allozyme and ISSR molecular markers[J].Heredity,2001,86(5):545-556.
[9]朱岩芳,祝水金,李永平,等.ISSR分子标记技术在植物种质资源研究中的应用[J].种子,2010,29(2):55-59.
[10]易克,徐向利,卢向阳,等.利用SSR和ISSR标记技术构建西瓜分子遗传图谱[J].湖南农业大学学报(自然科学版),2003,29(4):333-337.
[11]李冬梅.饲草型小黑麦的遗传图谱构建及草产量和抗锈病相关基因的QTL定位[D].兰州:甘肃农业大学,2016.
[12]Ruas P M,Ruas C F,Rampim L,et al.Genetic relationship in coffea species and parentage determination of interspecific hybrids using ISSR(Inter-Simple Sequence Repeat)markers[J].Genetics and Molecular Biology,2003,26(3):319-327.
[13]Aga E,Bekele E,Bryngelsson T.Inter-simple sequence repeat(ISSR)variation in forest coffee trees(Coffea arabica L.)populations from Ethiopia[J].Genetica,2005,124(2-3):213-221.
[14]Masumbuko L I,Bryngelsson T.Inter simple sequence repeat(ISSR)analysis of diploid coffee species and cultivated Coffea arabica L.from Tanzania[J].Genetic Resources and Crop Evolution,2006,53(2):357-366.
[15]Kumar S A,Sudisha J,Sreenath H L.Genetic relation of Coffea and Indian Psilanthus species as revealed through RAPD and ISSR markers[J].International Journal of Integrative Biology,2008,3(3):150-158.
[16]Tshilenge P,Nkongolo K K,Mehes M,et al.Genetic variation in Coffea canephora L.(Var.Robusta)accessions from the founder gene pool evaluated with ISSR and RAPD[J].African Journal of Biotechnology,2009,8(3):380-390.
[17]Kassahun T,Kim G,Endashaw B,et al.ISSR fingerprinting of Coffea arabica throughout Ethiopia reveals high variability in wild populations and distinguishes them from landraces[J].Plant Systematics and Evolution,2014,300(5):881-897.
[18]周华,张洪波,夏红云,等.咖啡种质资源多样性研究[J].中国热带农业,2015,5:48-52.
[19]黄丽芳,董云萍,王晓阳,等.利用RAPD标记分析咖啡种质资源的遗传多样性[J].热带作物学报,2014,35(12):2313-2319.
[20]黄丽芳,董云萍,王晓阳,等.云南咖啡资源遗传多样性的RAPD分析[J].中国热带农业,2017,5:48-52.
[21]黄丽芳,董云萍,王晓阳,等.咖啡种质资源DNA指纹图谱的构建[J].热带农业科学,2016,(36),12:48-52.
[22]黄丽芳,闫林,董云萍,等.咖啡叶片DNA提取方法的比较研究[J].热带农业科学,2011,(31),12:42-45.
[23]闫林,黄丽芳,谭乐和,等.咖啡ISSR与RAPD-PCR反应体系优化[J].热带作物学报,2012,33(5):854-859.
[24]王野,陈磊,白云,等.云杉矮槲寄生遗传多样性的ISSR分析[J].西北植物学报,2017,37(11):2153-2162.
[25]时圣明,潘明佳,王洁,等.分子鉴定技术在中药中的应用[J].中草药,2016,47(17):3121-3126.
[26]马克平.试论生物多样性的概念[J].生物多样性,1993,1(1):20-22.
[27]郑勇民,张旭业,徐秉良,等.美洲南瓜种质资源遗传多样性的ISSR分析[J].甘肃农业大学学报,2017,6:76-82.
[28]Masumbuko L I,Bryngelsson T,Mneney E E.Genetic diversity in Tanzanian Arabica coffee using random amplified polymorphic DNA(RAPD)markers[J].Hereditas,2010,139(1):56-63.
[29]Rotondi A,Magli M,Ricciolini C,et al.Morphological andmolecular analyses for the characterization of a group of Italyolive cuhivars[J].Euphytica,2003,132(2):129-137.
[30]陈海云,宁德鲁,李勇杰,等.59个油橄榄种质的ISSR分子鉴定[J].东北林业大学学报,2013,41(3):13-17.
[31]席春奕,唐茜,吴永胜,等.30份四川茶树种质资源遗传多样性与亲缘关系的SRAP分析[J].贵州农业科学,2013,41(2):6-9.