秦黑杨1号等美洲黑杨指纹图谱构建及遗传关系分析
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摘要
以西北农林科技大学渭河试验站的12个美洲黑杨无性系为材料,从国际杨树基因组委员会官方网站中提供的引物中选取25对引物对美洲黑杨进行指纹图谱构建,最终筛选出5对多态性丰富、扩增效率高、条带清晰的引物。5对引物共扩增条带24条,其中多态性条带23条,占95.8%,SSR引物多态性条带数4～8条,平均4.8条。对12份种质进行UPGMA聚类分析,品种遗传相似性系数在0.458 3～0.958 3,平均相似系数为0.666 0。研究利用5对引物(PMGC_2217、PMGC_2675、PMGC_2866、WPMS_14、WPMS_17)可将12份美洲黑杨种质完全区分开。研究结果可为美洲黑杨品种鉴定和知识产权保护提供理论依据。
Twelve Populus deltoides clones from Weihe Experimental Station of Northwest A&F University were used as materials,and 25 pairs of primers were selected from the official website of the International Poplar Genomics Commission to construct fingerprints of P.deltoides and eventually to screen out 5 pairs of rich polymorphism,highly amplified,and clear band primers.Five pairs of primers co-amplified 24 bands,including 23 polymorphic bands,accounting for 95.8%,and 4-8 polymorphic bands of SSR primers,with an average of 4.8.A UPGMA cluster analysis was performed on 12 germplasms.The genetic similarity coefficients of the varieties were between 0.458 3-0.958 3,and the average similarity coefficient was 0.666 0.In this study,using 5 pairs of primers(PMGC_2217,PMGC_2675,PMGC_2866,WPMS_14,WPMS_17) could completely distinguish 12 P.deltoides germplasms.The results of this study could provide a theoretical basis for the identification and protection of intellectual property of P.deltoides.
Twelve Populus deltoides clones from Weihe Experimental Station of Northwest A&F University were used as materials,and 25 pairs of primers were selected from the official website of the International Poplar Genomics Commission to construct fingerprints of P.deltoides and eventually to screen out 5 pairs of rich polymorphism,highly amplified,and clear band primers.Five pairs of primers co-amplified 24 bands,including 23 polymorphic bands,accounting for 95.8%,and 4-8 polymorphic bands of SSR primers,with an average of 4.8.A UPGMA cluster analysis was performed on 12 germplasms.The genetic similarity coefficients of the varieties were between 0.458 3-0.958 3,and the average similarity coefficient was 0.666 0.In this study,using 5 pairs of primers(PMGC_2217,PMGC_2675,PMGC_2866,WPMS_14,WPMS_17) could completely distinguish 12 P.deltoides germplasms.The results of this study could provide a theoretical basis for the identification and protection of intellectual property of P.deltoides.
引文
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[17] 王竹林,杨睿,刘联正,等.聚丙烯酰胺凝胶电泳银染的影响因素[J].实验室研究与探索,2011,30(9):15-17,113.WANG Z L,YANG R,LIU L Z,et al.Factors of silver staining of polyacrylamide gel electrophoresis[J].Research and Exploration in Laboratory,2011,30(9):15-17,113.(in Chinese)
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[2] 吕士行,徐锡增.美洲黑杨及其引种展望[J].南京林产工业学院学报,1980(3):13-21.LU S X,XU X Z.Populus deltoids and their prospect of being introduced from abroad[J].Journal of Nanjing Forestry University:Natural Sciences Edition,1980(3):13-21.(in Chinese)
[3] 符毓秦,刘玉媛,李均安,等.美洲黑杨杂种无性系——陕林3、4号杨的选育[J].陕西林业科技,1990,3(13):1-9,13.
[4] 王洁华,籍燕,杨少辉,等.速生优质北创杨新品系的选育[J].天津大学学报:自然科学与工程技术版,2014,47(10):914-918.WANG J H,JI Y,YANG S H,et al.Breeding of a fast-growing and high quality Beichuang variety[J].Journal of Tianjin University:Science and Technology,2014,47(10):914-918.(in Chinese)
[5] 苏晓华,丁昌俊,马常耕,等.我国杨树育种的研究进展及对策[J].林业科学研究,2010,23 (1):31-37.
[6] 黄秦军,苏晓华,张香华.SSR分子标记与林木遗传育种[J].世界林业研究,2002,15(3):14-21.HUANG Q J,SU X H,ZHANG X H.Microsatellite marker and its application in tree genetics and breeding[J].World Forestry Research,2002,15(3):14-21.
[7] 佘花娣,陈景堂,黄亚群,等.利用DNA指纹图谱进行农作物品种鉴定的研究进展[J].河北农业大学学报,2003,26(增刊1):28-30,33.SHE H D,CHEN J T,HUANG Y Q,et al.Progress on variety identification of crop using DNA fingerprinting[J].Journal of Agricultural University of Hebei,2003,26(Supp.1):28-30,33.(in Chinese)
[8] 吴敏,杜红岩,乌云塔娜,等.杜仲基因组微卫星特征及SSR标记开发[J].林业科学研究,2015,28(3):387-393.
[9] 齐建勋,郝艳宾,朱艳,等.核桃属种质资源的EST-SSR标记研究[J].园艺学报,2011,38(3):441-448.QI J X,HAO Y B,ZHU Y,et al.Studies on germplasm of Juglans by EST-SSR markers[J].Acta Horticulturae Sinica,2011,38(3):441-448.(in Chinese)
[10] DAYANANDAN S,RAJORA O P,BAWA K S.Isolation and characterization of microsatellites in trembling aspen (Populus tremuloides)[J].Theoretical & Applied Genetics,1998,96(6-7):950-956.
[11] 藕丹,樊军锋,周永学,等.美洲黑杨×青杨派杂种无性系SSR指纹图谱的构建与遗传差异性分析[J].西北林学院学报,2017,32(2):112-116.OU D,FAN J F,ZHOU Y X,et al.Construction of fingerprint and analysis of genetic difference for hybrid clones of Populus deltoides×Section tacamahaca using SSR marker[J].Journal of Northwest Forestry University,2017,32(2):112-116.(in Chinese)
[12] 韩志校,张军,左力辉,等.基于SSR和ITS标记的杨树多态性及聚类分析[J].中国农业科技导报,2017,19(7):36-43.
[13] 李薇,刘春英,郭娟,等.3个美洲黑杨新无性系的SSR分析及鉴定[J].西北林学院学报,2014,29(1):73-77.LI W,LIU C Y,GUO J,et al.Analysis and identification of three new clones of Populus deltoides based on SSR molecular markers[J].Journal of Northwest Forestry University,2014,29(1):73-77.(in Chinese)
[14] 李金璐,王硕,于婧,等.一种改良的植物DNA提取方法[J].植物学报,2013,48(1):72-78.LI J L,WANG S,YU J,et al.A modified CTAB protocol for plant DNA extraction[J].Chinese Bulletin of Botany,2013,48(1):72-78.(in Chinese)
[15] 樊蓉,樊军锋,李周岐.9个白杨品种SSR指纹图谱构建及遗传关系的研究[J].西北林学院学报,2015,30(3):76-80.FAN R,FAN J F,LI Z Q,Fingerprinting and genetic relatedness of 9 varieties in Populus L.sect.Populus using SSR markers[J].Journal of Northwest Forestry University,2015,30(3):76-80.(in Chinese)
[16] 韩志校,张军,左力辉,等.3个不同派别杨树资源遗传多样性的SSR分析[J].河南农业科学,2017,46(4):99-103.
[17] 王竹林,杨睿,刘联正,等.聚丙烯酰胺凝胶电泳银染的影响因素[J].实验室研究与探索,2011,30(9):15-17,113.WANG Z L,YANG R,LIU L Z,et al.Factors of silver staining of polyacrylamide gel electrophoresis[J].Research and Exploration in Laboratory,2011,30(9):15-17,113.(in Chinese)
[18] 郭娟.美洲黑杨5个优良品种EST-SSR和SRAP指纹图谱构建[D].杨陵:西北农林科技大学,2014.
[19] 贾会霞,姬慧娟,胡建军,等.杨树新品种的SSR指纹图谱构建和倍性检测[J].林业科学,2015,51(2):69-79.JIA H X,JI H J,HU J J,et al.Fingerprints of SSR markers and ploidyd detection for new Populus varieties[J].Scientia Silvae Sinicae,2015,51(2):69-79.(in Chinese)
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