大葱种质资源亲缘关系的ISSR分析
|
摘要
研究不同性状大葱种质资源的亲缘关系,并构建DNA指纹图谱,为大葱种质资源分类、品种鉴定、新品种选育提供参考依据。以具有不同性状的71份大葱种质资源为材料,用ISSR分子标记分析大葱种质资源的遗传多样性和品种亲缘关系。用筛选出的14条扩增条带清晰、多态性较好的ISSR引物对71份种质资源进行ISSR-PCR扩增。共得到97条清晰的谱带,结果每个引物平均扩增出6.93条谱带,多态性较好的谱带60条,多态性条带比率为61.9%。利用ISSR技术能有效地从分子水平揭示出大葱种质资源的遗传多样性和品种间的亲缘关系,并能有效应用于大葱品种指纹图谱的构建。本研究结果为大葱种质资源的进一步开发利用提供理论依据。
This study was carried out to investigate the genetic relationship of different characters of welsh onion germplasm resources and to construct DNA fingerprint, provide reference for classification, variety identification and new variety breeding of welsh onion germplasm resources. Genetic diversity and genetic relationship of 71 onion germplasm resources with different characters were analyzed by ISSR markers. The ISSR-PCR amplification of 71 germplasm resources was carried out by 14 ISSR primers with clear and polymorphic amplification bands. A total of 97 clear bands were obtained, with an average of 6.93 bands per primer. 60 of them were polymorphic bands, with the polymorphic bands ratio of 61.9%. ISSR technology could effectively reveal genetic diversity and genetic relationship among varieties of welsh onion germplasm resources at the molecular level, and could be effectively applied to the construction of fingerprint of welsh onion varieties. The results of this study could provide theoretical basis for further exploration and utilization of welsh onion germplasm resources.
This study was carried out to investigate the genetic relationship of different characters of welsh onion germplasm resources and to construct DNA fingerprint, provide reference for classification, variety identification and new variety breeding of welsh onion germplasm resources. Genetic diversity and genetic relationship of 71 onion germplasm resources with different characters were analyzed by ISSR markers. The ISSR-PCR amplification of 71 germplasm resources was carried out by 14 ISSR primers with clear and polymorphic amplification bands. A total of 97 clear bands were obtained, with an average of 6.93 bands per primer. 60 of them were polymorphic bands, with the polymorphic bands ratio of 61.9%. ISSR technology could effectively reveal genetic diversity and genetic relationship among varieties of welsh onion germplasm resources at the molecular level, and could be effectively applied to the construction of fingerprint of welsh onion varieties. The results of this study could provide theoretical basis for further exploration and utilization of welsh onion germplasm resources.
引文
Dong H.Y.,Ji K.S.,Hou B.X.,and Zhao H.B.,2014,Genetic relatives analysis of 41 Loropetalum chinense var.rubrum cultivars by ISSR markers,Yuanyi Xuebao(Acta Horticulturae Sinica),41(2):365-374(董海燕,季孔庶,侯伯鑫,赵宏波,2014,基于ISSR标记的红花檵木品种亲缘关系分析,园艺学报,41(2):365-374)
Gao L.M.,Dong F.,Huo Y.M.,Liu B.J.,Liao J.,and Chen Y.Q.,2013,Development of SCAR marker identifying the cytoplasmic male sterility gene in bunching onion(Allium fistulosum L.),Yuanyi Xuebao(Acta Horticulturae Sinica),40(7):1382-1388(高莉敏,董飞,霍雨猛,刘冰江,缪军,陈运起,2013,大葱细胞质雄性不育基因的SCAR标记开发,园艺学报,40(7):1382-1388)
Hu X.H.,Li J.J.,Wang Y.,and Huang S.X.,2014,Reaction system optimization and primer screening of ISSR fingerprint map for Camellia cultivars,Beifang Yuanyi(Northern Horticulture),(22):84-88(胡兴华,李景剑,王燕,黄仕训,2014,茶花品种ISSR指纹图谱反应体系建立与优化,北方园艺,(22):84-88)
Jia J.X.,and Tao C.G.,2008,RAPD analysis of genetic relationship among some welsh onion cultivars,Zhongguo Shucai(China Vegetables),1(5):23-26(贾俊香,陶承光,2008,大葱部分种质资源亲缘关系的RAPD分析,中国蔬菜,1(5):23-26)
Jia J.X.,Cui L.W.,Li N.,and Jiang B.B.,2017,Establishment and optimization of ISSR reaction system on welsh onion,Beifang Yuanyi(Northern Horticulture),(12):92-96(贾俊香,崔连伟,李娜,姜滨滨,2017,大葱ISSR反应体系的建立及优化,北方园艺,(12):92-96)
Jin X.,Zhou X.G.,Liu SW.,Liu SQ.,and Jia T.J.,2013,ISSRmarkers of 48 tillering onion(Allium cepa var.aggregatum)germplasms genetic diversity and analysis of their agronom ic traits,Zhongguo Shucai(China Vegetables),(2):27-34(金雪,周新刚,刘守伟,刘淑芹,吴凤芝,2013,48份分蘖洋葱种质资源遗传多样性的ISSR标记和农艺性状分析,中国蔬菜,(2):27-34)
Liu B.H.,Deng C.L.,Chen C.W.,Deng G.Y.,Ding P.,Niu Y.,Tang Y.,and Fu H.M.,2015,Genetic analysis of local Citrus germplasm resources in Guangxi by ISSR markers,Guoshu Xuebao(Journal of Fruit Science),32(6):1001-1006(刘冰浩,邓崇岭,陈传武,邓光宙,丁萍,牛英,唐艳,付慧敏,2015,广西地方柑橘资源的ISSR分析,果树学报,32(6):1001-1006)
Liu J.,Liao K.,Mansuer N.,Sun Q.,Liu H.,and Jia Y.,2015,Analysis of genetic diversity and construction of DNA fingerprint database of Xinjiang apricot varieties(lines),Zhongguo Nongye Kexue(Scientia Agricultura Sinica),48(4):748-758(刘娟,廖康,曼苏尔·那斯尔,孙琪,刘欢,贾杨,2015,新疆杏品种(系)遗传多样性分析及DNA指纹图谱库构建,中国农业科学,48(4):748-758)
Ohara T.,Song Y.S.,Tsukazaki H.,Wako T.,Nunome T.,and Kojima A.,2005,Genetic mapping of AFLP markers in Japanese bunching onion(Allium fistulosum),Euphytica,144(3):255-263
Parvathaneni R.K.,Natesan S.,Devaraj A.A.,Muthuraja R.,Venkatachalam R.,Subramani A.P.,and Laxmanan P.,2011,Fingerprinting in cucumber and melon(Cucumis spp.)genotypes using morphological and ISSR markers,J.Crop Sci.Biotech.,14(1):39-43
Peng J.Q.,Cao F.X.,Cao J.W.,Dong X.J.,Liu Z.M.,Ren R.,Xiong Y.L.,and Zhang P.H.,2016,Genetic diversity and fingerprint construction of varieties of Hydrangea macrophylla by ISSR markers,Zhongnan Linye Keji Daxue Xuebao(Journal of Central South University of Forestry&Technology),36(12):115-120(彭继庆,曹福祥,曹基武,董旭杰,刘志明,任锐,熊亚丽,张鹏鸿,2016,绣球花品种遗传多样性的ISSR分析及指纹图谱构建,中南林业科技大学学报,36(12):115-120)
Tsukazaki H.,Yamashita K.I.,Kojima A.,and Wako T.,2009,SSR-tagged breeding s cheme for allogamous crops:a trial in bunching onion(Allium fistulosum),Euphytica,169(3):327-334
Wang C.,Zhao H.,Zhang L.Y.,Pei Y.X.,and Wang Y.Q.,2015,RFLP analysis of mitochondrial genomes of cytoplasmic male sterile lines and maintainer lines in welsh onion(Allium fistulosum L.),Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),23(7):888-893(王婵,赵泓,张丽英,裴雁曦,王永勤,2015,大葱细胞质雄性不育系及保持系线粒体基因组RFLP分析,农业生物技术学报,23(7):888-893)
Yang X.,Liu F.,Zhang Y.,Cheng Y.F.,Xue L.B.,and Chen X.H.,2016,Study on the genetic diversity of eggplant germplasm with SSR markers,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(12):3450-3457(杨旭,刘飞,张宇,成玉富,薛林宝,陈学好,2016,利用SSR标记研究茄子种质资源遗传多样性,基因组学与应用生物学,35(12):3450-3457)
Yin L.Q.,Peng Y.Q.,Fu S.H.,Yang J.,Chen T.,Huang M.,Yu Q.,Wei X.Y.,and Niu Y.Z.,2016,Genetic diversity and DNA fingerprint of pigmented potato(Solanumtuberosum L.)based on ISSR markers,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),39(1):20-25(殷丽琴,彭云强,付绍红,杨进,陈涛,黄敏,余勤,韦献雅,牛应泽,2016,基于ISSR标记的彩色马铃薯遗传多样性分析及指纹图谱构建,西南农业学报,39(1):20-25)
Zhang M.,Li Y.L.,W Z.,Yin S.H.,and Liu H.M.,2012,The primary exploring of genetic diversity of leek varieties based on ISSR markers,Xibei Nongye Xuebao(Acta Agriculture Boreali-occidentalis Sinica),21(1):146-150(张明,李延龙,王贞,尹守恒,刘宏敏,2012,基于ISSR标记的韭菜种质资源遗传多样性初探,西北农业学报,21(1):146-150)
Zhang X.F.,Li.X.,Cui L.N.,Zhou C.J.,Li J.,and Long Y.C.,2015,Genetic diversity analysis of Cercospora spp.by ISSRin China,Zhiwu Baohu Xuebao(Journal of Plant Protection),42(6):908-913(张小飞,李晓,崔丽娜,邹成佳,李菁,龙永昌,2015,我国玉米灰斑病菌遗传多样性的ISSR分析,植物保护学报,42(6):908-913)
Zhao Y.X.,Gao Q.,Sun S.Y.,Wu Y.X.,Zhang R.X.,and Hao W.S.,2016,ISSR analysis on genetic diversity of different varieties(lines)in Solanum tuberosum L.,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(12):3434-3435(赵永秀,高茜,孙淑英,吴玉仙,张瑞霞,郝文胜,2016,马铃薯不同品种(系)遗传多样性的ISSR分析,分子植物育种,14(12):3434-3435)
Gao L.M.,Dong F.,Huo Y.M.,Liu B.J.,Liao J.,and Chen Y.Q.,2013,Development of SCAR marker identifying the cytoplasmic male sterility gene in bunching onion(Allium fistulosum L.),Yuanyi Xuebao(Acta Horticulturae Sinica),40(7):1382-1388(高莉敏,董飞,霍雨猛,刘冰江,缪军,陈运起,2013,大葱细胞质雄性不育基因的SCAR标记开发,园艺学报,40(7):1382-1388)
Hu X.H.,Li J.J.,Wang Y.,and Huang S.X.,2014,Reaction system optimization and primer screening of ISSR fingerprint map for Camellia cultivars,Beifang Yuanyi(Northern Horticulture),(22):84-88(胡兴华,李景剑,王燕,黄仕训,2014,茶花品种ISSR指纹图谱反应体系建立与优化,北方园艺,(22):84-88)
Jia J.X.,and Tao C.G.,2008,RAPD analysis of genetic relationship among some welsh onion cultivars,Zhongguo Shucai(China Vegetables),1(5):23-26(贾俊香,陶承光,2008,大葱部分种质资源亲缘关系的RAPD分析,中国蔬菜,1(5):23-26)
Jia J.X.,Cui L.W.,Li N.,and Jiang B.B.,2017,Establishment and optimization of ISSR reaction system on welsh onion,Beifang Yuanyi(Northern Horticulture),(12):92-96(贾俊香,崔连伟,李娜,姜滨滨,2017,大葱ISSR反应体系的建立及优化,北方园艺,(12):92-96)
Jin X.,Zhou X.G.,Liu SW.,Liu SQ.,and Jia T.J.,2013,ISSRmarkers of 48 tillering onion(Allium cepa var.aggregatum)germplasms genetic diversity and analysis of their agronom ic traits,Zhongguo Shucai(China Vegetables),(2):27-34(金雪,周新刚,刘守伟,刘淑芹,吴凤芝,2013,48份分蘖洋葱种质资源遗传多样性的ISSR标记和农艺性状分析,中国蔬菜,(2):27-34)
Liu B.H.,Deng C.L.,Chen C.W.,Deng G.Y.,Ding P.,Niu Y.,Tang Y.,and Fu H.M.,2015,Genetic analysis of local Citrus germplasm resources in Guangxi by ISSR markers,Guoshu Xuebao(Journal of Fruit Science),32(6):1001-1006(刘冰浩,邓崇岭,陈传武,邓光宙,丁萍,牛英,唐艳,付慧敏,2015,广西地方柑橘资源的ISSR分析,果树学报,32(6):1001-1006)
Liu J.,Liao K.,Mansuer N.,Sun Q.,Liu H.,and Jia Y.,2015,Analysis of genetic diversity and construction of DNA fingerprint database of Xinjiang apricot varieties(lines),Zhongguo Nongye Kexue(Scientia Agricultura Sinica),48(4):748-758(刘娟,廖康,曼苏尔·那斯尔,孙琪,刘欢,贾杨,2015,新疆杏品种(系)遗传多样性分析及DNA指纹图谱库构建,中国农业科学,48(4):748-758)
Ohara T.,Song Y.S.,Tsukazaki H.,Wako T.,Nunome T.,and Kojima A.,2005,Genetic mapping of AFLP markers in Japanese bunching onion(Allium fistulosum),Euphytica,144(3):255-263
Parvathaneni R.K.,Natesan S.,Devaraj A.A.,Muthuraja R.,Venkatachalam R.,Subramani A.P.,and Laxmanan P.,2011,Fingerprinting in cucumber and melon(Cucumis spp.)genotypes using morphological and ISSR markers,J.Crop Sci.Biotech.,14(1):39-43
Peng J.Q.,Cao F.X.,Cao J.W.,Dong X.J.,Liu Z.M.,Ren R.,Xiong Y.L.,and Zhang P.H.,2016,Genetic diversity and fingerprint construction of varieties of Hydrangea macrophylla by ISSR markers,Zhongnan Linye Keji Daxue Xuebao(Journal of Central South University of Forestry&Technology),36(12):115-120(彭继庆,曹福祥,曹基武,董旭杰,刘志明,任锐,熊亚丽,张鹏鸿,2016,绣球花品种遗传多样性的ISSR分析及指纹图谱构建,中南林业科技大学学报,36(12):115-120)
Tsukazaki H.,Yamashita K.I.,Kojima A.,and Wako T.,2009,SSR-tagged breeding s cheme for allogamous crops:a trial in bunching onion(Allium fistulosum),Euphytica,169(3):327-334
Wang C.,Zhao H.,Zhang L.Y.,Pei Y.X.,and Wang Y.Q.,2015,RFLP analysis of mitochondrial genomes of cytoplasmic male sterile lines and maintainer lines in welsh onion(Allium fistulosum L.),Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),23(7):888-893(王婵,赵泓,张丽英,裴雁曦,王永勤,2015,大葱细胞质雄性不育系及保持系线粒体基因组RFLP分析,农业生物技术学报,23(7):888-893)
Yang X.,Liu F.,Zhang Y.,Cheng Y.F.,Xue L.B.,and Chen X.H.,2016,Study on the genetic diversity of eggplant germplasm with SSR markers,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(12):3450-3457(杨旭,刘飞,张宇,成玉富,薛林宝,陈学好,2016,利用SSR标记研究茄子种质资源遗传多样性,基因组学与应用生物学,35(12):3450-3457)
Yin L.Q.,Peng Y.Q.,Fu S.H.,Yang J.,Chen T.,Huang M.,Yu Q.,Wei X.Y.,and Niu Y.Z.,2016,Genetic diversity and DNA fingerprint of pigmented potato(Solanumtuberosum L.)based on ISSR markers,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),39(1):20-25(殷丽琴,彭云强,付绍红,杨进,陈涛,黄敏,余勤,韦献雅,牛应泽,2016,基于ISSR标记的彩色马铃薯遗传多样性分析及指纹图谱构建,西南农业学报,39(1):20-25)
Zhang M.,Li Y.L.,W Z.,Yin S.H.,and Liu H.M.,2012,The primary exploring of genetic diversity of leek varieties based on ISSR markers,Xibei Nongye Xuebao(Acta Agriculture Boreali-occidentalis Sinica),21(1):146-150(张明,李延龙,王贞,尹守恒,刘宏敏,2012,基于ISSR标记的韭菜种质资源遗传多样性初探,西北农业学报,21(1):146-150)
Zhang X.F.,Li.X.,Cui L.N.,Zhou C.J.,Li J.,and Long Y.C.,2015,Genetic diversity analysis of Cercospora spp.by ISSRin China,Zhiwu Baohu Xuebao(Journal of Plant Protection),42(6):908-913(张小飞,李晓,崔丽娜,邹成佳,李菁,龙永昌,2015,我国玉米灰斑病菌遗传多样性的ISSR分析,植物保护学报,42(6):908-913)
Zhao Y.X.,Gao Q.,Sun S.Y.,Wu Y.X.,Zhang R.X.,and Hao W.S.,2016,ISSR analysis on genetic diversity of different varieties(lines)in Solanum tuberosum L.,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(12):3434-3435(赵永秀,高茜,孙淑英,吴玉仙,张瑞霞,郝文胜,2016,马铃薯不同品种(系)遗传多样性的ISSR分析,分子植物育种,14(12):3434-3435)