毛白杨多态SSR引物库和种质资源指纹图谱库构建
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摘要
【目的】针对毛白杨优树种质资源形态学差异小、不易区分等问题,利用多态SSR引物构建优树种质资源指纹图谱,为毛白杨种质资源管理、新品种选育、知识产权保护等提供参考。【方法】本研究以山东冠县毛白杨种质资源库469个优树无性系为对象,从毛白杨SSR多态引物筛选入手,通过荧光SSR引物PCR扩增和毛细管电泳仪检测筛选SSR引物,并构建指纹图谱。【结果】在2 317对SSR引物中,共计得到清晰、特异、多态、稳定扩增的SSR引物406对,这些SSR引物在19条染色体上的分布数量介于3～39对之间,利用BLAST比对分析可将其中389对SSR引物定位到毛果杨基因组,构建了毛白杨优树种质资源多态SSR引物库。应用多态性较高的SSR核心引物,以及特异的SSR辅助引物相结合的技术策略,筛选出25对多态性较高的SSR核心引物,以及13对特异的SSR辅助引物,构建了毛白杨种质资源库内469个优良无性系指纹图谱库,并完成了指纹图谱的QR编码。【结论】本研究成功构建了毛白杨种质资源多态SSR引物库和优树无性系指纹图谱库,对与毛白杨类似的林木种质资源遗传变异研究以及品种鉴定、系谱分析等具有重要意义。
[Objective] Aimed at the problems that the morphological differences of superior tree germplasm resources of Populus tomentosa were small and difficult to distinguish, polymorphic SSR primers were used to construct the fingerprint of superior tree germplasm resources, provide references for the germplasm resource management, breeding of new cultivar, intellectual property protection, etc.[Method] In this study, 469 superior trees from Populus tomentosa germplasm resource bank in Guanxian County, Shandong Province of eastern China were used as material. Firstly, the SSR polymorphic primers were screened by PCR amplification and capillary electrophoresis detection, and then the fingerprint was constructed. [Result] In 2 317 pairs of SSR primers, a total of 406 pairs of SSR primers with clear, specific,polymorphic and stable amplification were obtained. These SSR primers distributed on 19 chromosomes and ranged in 3-39 pairs. Further, 389 pairs of SSR primers were located into the genome of P. trichocarpa based on BLAST comparison analysis, and a polymorphic SSR primer library of Populus tomentosa germplasm resources was constructed. On this basis, 25 pairs of SSR primers with high polymorphism and13 pairs of specific SSR-assisted primers were used to construct the fingerprint of 469 superior trees in the germplasm resource bank of Populus tomentosa, and the QR coding of the fingerprint was completed.[Conclusion] This study successfully constructed the polymorphic SSR primer library and the superior tree fingerprint library of Populus tomentosa germplasm resources, and be of great significance to the study of genetic variation, variety identification and pedigree analysis of Populus tomentosa.
[Objective] Aimed at the problems that the morphological differences of superior tree germplasm resources of Populus tomentosa were small and difficult to distinguish, polymorphic SSR primers were used to construct the fingerprint of superior tree germplasm resources, provide references for the germplasm resource management, breeding of new cultivar, intellectual property protection, etc.[Method] In this study, 469 superior trees from Populus tomentosa germplasm resource bank in Guanxian County, Shandong Province of eastern China were used as material. Firstly, the SSR polymorphic primers were screened by PCR amplification and capillary electrophoresis detection, and then the fingerprint was constructed. [Result] In 2 317 pairs of SSR primers, a total of 406 pairs of SSR primers with clear, specific,polymorphic and stable amplification were obtained. These SSR primers distributed on 19 chromosomes and ranged in 3-39 pairs. Further, 389 pairs of SSR primers were located into the genome of P. trichocarpa based on BLAST comparison analysis, and a polymorphic SSR primer library of Populus tomentosa germplasm resources was constructed. On this basis, 25 pairs of SSR primers with high polymorphism and13 pairs of specific SSR-assisted primers were used to construct the fingerprint of 469 superior trees in the germplasm resource bank of Populus tomentosa, and the QR coding of the fingerprint was completed.[Conclusion] This study successfully constructed the polymorphic SSR primer library and the superior tree fingerprint library of Populus tomentosa germplasm resources, and be of great significance to the study of genetic variation, variety identification and pedigree analysis of Populus tomentosa.
引文
[1]朱之悌,康向阳,张志毅.毛白杨天然三倍体选种研究[J].林业科学,1998,34(4):22-31.Zhu Z T,Kang X Y,Zhang Z Y.Studies on selection of natural triploids of Populus tomentosa[J].Scientia Silvae Sinicae,1998,34(4):22-31.
[2]朱之悌.毛白杨遗传改良[M].北京：中国林业出版社,2006.Zhu Z T.Genetic improvement of Populus tomentosa Carr.[M].Beijing:China Forestry Publishing House,2006.
[3]Guichoux E,Lagache L,Wgner S,et al.Current trends in microsatellite genotyping[J].Molecular Ecology Resources,2011,11(4):591-611.
[4]Powell W,Machray G C,Provan J.Polymorphism revealed by simple sequence repeats[J].Trends in Plant Science,1996,7(1):215-222.
[5]Dayanandan S,Rajora O P,Bawa K S.Isolation and characterization of microsatellites in trembling aspen(Populus tremuloides)[J].Theoretical and Applied Genetics,1998,96(6/7):950-956.
[6]Yin T M,Difazio S P,Gunter L E,et al.Large-scale heterospecific segregation distortion in Populus revealed by a dense genetic map[J].Theoretical and Applied Genetics,2004,109(3):451-463.
[7]Liu H L,Yang W X,Hou J,et al.Genetic identification of 43 elite clonal accessions of Populus deltoides by SSR fingerprinting[J].Canadian Journal of Plant Science,1918,96(3):494-502.
[8]Han Z Q,Gao P,Geng X N,et al.Identification of the male parent of superior half-sib Populus tomentosa individuals based on SSRmarkers[J/OL].Molecular Breeding,2017,37(12):155[2019-01-13].https://doi.org/10.1007/s11032-017-0754-1.
[9]白凤莹,曾青青,康宁,等.毛白杨基因库优树倍性检测及性状对比分析[J].北京林业大学学报,2015,37(4):113-118.Bai F Y,Zeng Q Q,Kang N,et al.Ploidy level and contrast analysis of the traits for superior trees of Populus tomentosa Carr.in gene pool[J].Journal of Beijing Forestry University,2015,37(4):113-118.
[10]Schuelke,M.An economic method for the fluorescent labeling of PCR fragments[J].Nature Biotechnology,2000,18:233-234.
[11]Hulce D,Li X,Snyder-Leiby T,et al.GeneMarker?genotyping software:tools to increase the statistical power of DNA fragment analysis[J].Journal of Biomolecular Techniques,2011,22(Suppl.):35.
[12]Yeh F C,Yang R C,Boyle T.POPGENE Version 1.32:Microsoft Windows-based freeware for populations genetic analysis[Z].Edmonton:University of Alberta,1999.
[13]马红勃,许旭明,韦新宇,等.基于SSR标记的福建省若干水稻品种DNA指纹图谱构建及遗传多样性分析[J].福建农业学报,2010,25(1):33-38.Ma H B,Xu X M,Wei X Y,et al.DNA fingerprints and genetic diversity analysis based on SSR markers for rice cultivars in Fujian[J].Fujian Journal of Agricultural Sciences,2010,25(1):33-38.
[14]王静毅,陈业渊,黄秉智,等.部分香蕉品种SSR指纹图谱的构建[J].果树学报,2009,26(5):733-738.Wang J Y,Chen Y Y,Huang B Z,et al.Establishment of fingerprinting for bananas(Musa nana)by SSR marker[J].Journal of Fruit Science,2009,26(5):733-738.
[15]Powell W,Machray G C,Provan J.Polymorphism revealed by simple sequence repeat[J].Trends in Plant Science,1996,1(7):215-222.
[16]陈琛,张兴桃,程斐,等.秋甘蓝品种的SSR指纹图谱的构建[J].园艺学报,2011,38(1):159-164.Chen C,Zhang X T,Cheng F,et al.Establishment of SSRfingerprinting on autumn cabbage hybrids and their parents[J].Acta Horticulturae Sinica,2011,38(1):159-164.
[17]刘本英,王丽鸳,李友勇,等.ISSR标记鉴别云南茶树种质资源的研究[J].茶叶科学,2009,29(5):355-364.Liu B Y,Wang L Y,Li Y Y,et al.ISSR markers for discriminating tea germplasm resources from Yunnan Province[J].Journal of Tea Science,2009,29(5):355-364.
[2]朱之悌.毛白杨遗传改良[M].北京：中国林业出版社,2006.Zhu Z T.Genetic improvement of Populus tomentosa Carr.[M].Beijing:China Forestry Publishing House,2006.
[3]Guichoux E,Lagache L,Wgner S,et al.Current trends in microsatellite genotyping[J].Molecular Ecology Resources,2011,11(4):591-611.
[4]Powell W,Machray G C,Provan J.Polymorphism revealed by simple sequence repeats[J].Trends in Plant Science,1996,7(1):215-222.
[5]Dayanandan S,Rajora O P,Bawa K S.Isolation and characterization of microsatellites in trembling aspen(Populus tremuloides)[J].Theoretical and Applied Genetics,1998,96(6/7):950-956.
[6]Yin T M,Difazio S P,Gunter L E,et al.Large-scale heterospecific segregation distortion in Populus revealed by a dense genetic map[J].Theoretical and Applied Genetics,2004,109(3):451-463.
[7]Liu H L,Yang W X,Hou J,et al.Genetic identification of 43 elite clonal accessions of Populus deltoides by SSR fingerprinting[J].Canadian Journal of Plant Science,1918,96(3):494-502.
[8]Han Z Q,Gao P,Geng X N,et al.Identification of the male parent of superior half-sib Populus tomentosa individuals based on SSRmarkers[J/OL].Molecular Breeding,2017,37(12):155[2019-01-13].https://doi.org/10.1007/s11032-017-0754-1.
[9]白凤莹,曾青青,康宁,等.毛白杨基因库优树倍性检测及性状对比分析[J].北京林业大学学报,2015,37(4):113-118.Bai F Y,Zeng Q Q,Kang N,et al.Ploidy level and contrast analysis of the traits for superior trees of Populus tomentosa Carr.in gene pool[J].Journal of Beijing Forestry University,2015,37(4):113-118.
[10]Schuelke,M.An economic method for the fluorescent labeling of PCR fragments[J].Nature Biotechnology,2000,18:233-234.
[11]Hulce D,Li X,Snyder-Leiby T,et al.GeneMarker?genotyping software:tools to increase the statistical power of DNA fragment analysis[J].Journal of Biomolecular Techniques,2011,22(Suppl.):35.
[12]Yeh F C,Yang R C,Boyle T.POPGENE Version 1.32:Microsoft Windows-based freeware for populations genetic analysis[Z].Edmonton:University of Alberta,1999.
[13]马红勃,许旭明,韦新宇,等.基于SSR标记的福建省若干水稻品种DNA指纹图谱构建及遗传多样性分析[J].福建农业学报,2010,25(1):33-38.Ma H B,Xu X M,Wei X Y,et al.DNA fingerprints and genetic diversity analysis based on SSR markers for rice cultivars in Fujian[J].Fujian Journal of Agricultural Sciences,2010,25(1):33-38.
[14]王静毅,陈业渊,黄秉智,等.部分香蕉品种SSR指纹图谱的构建[J].果树学报,2009,26(5):733-738.Wang J Y,Chen Y Y,Huang B Z,et al.Establishment of fingerprinting for bananas(Musa nana)by SSR marker[J].Journal of Fruit Science,2009,26(5):733-738.
[15]Powell W,Machray G C,Provan J.Polymorphism revealed by simple sequence repeat[J].Trends in Plant Science,1996,1(7):215-222.
[16]陈琛,张兴桃,程斐,等.秋甘蓝品种的SSR指纹图谱的构建[J].园艺学报,2011,38(1):159-164.Chen C,Zhang X T,Cheng F,et al.Establishment of SSRfingerprinting on autumn cabbage hybrids and their parents[J].Acta Horticulturae Sinica,2011,38(1):159-164.
[17]刘本英,王丽鸳,李友勇,等.ISSR标记鉴别云南茶树种质资源的研究[J].茶叶科学,2009,29(5):355-364.Liu B Y,Wang L Y,Li Y Y,et al.ISSR markers for discriminating tea germplasm resources from Yunnan Province[J].Journal of Tea Science,2009,29(5):355-364.
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