油橄榄×尖叶木樨榄种间杂交种真实性的SSR鉴定
|
摘要
油橄榄种间杂交可拓宽现有油橄榄种质资源的遗传基础,其杂种真实性鉴定可为种间杂交技术改良和种质利用提供技术条件。云南省林业科学院以佛奥(Olea europaea cv. Frantoio)作母本、尖叶木樨榄(O.cuspidata)作父本,成功杂交育得子代。本研究利用14个在亲本间存在差异的油橄榄SSR标记,对3个种间杂交子代和1个疑似杂交种"欧莱雅"的杂种真实性进行了鉴定。结果表明,1个SSR标记(EMO90)仅在佛奥和杂交子代中有SSR-PCR扩增产物,而在尖叶木樨榄中无产物,表现为显性标记;其余13个SSR标记(Oli23,Oli26, GAPU01, GAPU59, GAPU71, DCA05, DCA07, DCA08, DCA15, DCA18, EMO2, EMO3, UDO09)为共显性标记,其中3个SSR标记(GAPU71, EMO2和EMO3)能够将子代杂合而亲本纯合的基因型直观进行呈现,明确证实了杂交种的真实性。本研究为今后油橄榄远缘杂交育种时杂种的早期鉴定提供了技术支持。
Interspecific hybrid ization of olive can broaden the genetic basis of the existing olive germplasm resources, and hybrid's authenticity identification can provide technical support for the improvement of interspecific hybridization technology and the utilization of elite germplasm. Yunnan Academy of Forestry had bred a few varieties of olive by crossbreeding of Olea europaea cv. Frantoio as the female parent and O. cuspidata as the male parent. In this study, the authenticities of three hybrids and one suspected hybrid "Oulaiya" were identified by using 14 polymorphic olive SSR markers. The results showed that one SSR marker(EMO90) was a dominant marker, only had SSR-PCR amplification product in Frantoio, while no product was found in O.cuspidate. The remaining 13 SSR markers(Oli23, Oli26, GAPU01, GAPU59, GAPU71, DCA05, DCA07, DCA08,DCA15, DCA18, EMO2, EMO3, UDO09) were codominant markers. Among them, three SSR markers(GAPU71,EMO2 and EMO3) could visually present the different genotypes among parents(homozygous types) and hybrids(heterozygous types), and clearly prove the authenticity of hybrids. The results of this study could provide technique support for the early-stage evaluation of hybrid in the olive distant crossbreeding.
Interspecific hybrid ization of olive can broaden the genetic basis of the existing olive germplasm resources, and hybrid's authenticity identification can provide technical support for the improvement of interspecific hybridization technology and the utilization of elite germplasm. Yunnan Academy of Forestry had bred a few varieties of olive by crossbreeding of Olea europaea cv. Frantoio as the female parent and O. cuspidata as the male parent. In this study, the authenticities of three hybrids and one suspected hybrid "Oulaiya" were identified by using 14 polymorphic olive SSR markers. The results showed that one SSR marker(EMO90) was a dominant marker, only had SSR-PCR amplification product in Frantoio, while no product was found in O.cuspidate. The remaining 13 SSR markers(Oli23, Oli26, GAPU01, GAPU59, GAPU71, DCA05, DCA07, DCA08,DCA15, DCA18, EMO2, EMO3, UDO09) were codominant markers. Among them, three SSR markers(GAPU71,EMO2 and EMO3) could visually present the different genotypes among parents(homozygous types) and hybrids(heterozygous types), and clearly prove the authenticity of hybrids. The results of this study could provide technique support for the early-stage evaluation of hybrid in the olive distant crossbreeding.
引文
Baldoni L.,Cultrera N.G.,Mariotti R.,Ricciolini C.,Arcioni S.,Vendramin G.G.,Buonamici A.,Pordeddu A.,Sarri V.,Ojeda M.A.,Trujillo I.,Rallo L.,Belaj A.,Perri E.,Salimonti A.,Muzzalupo I.,Casagrande A.,Lain O.,Messina R.,and Testolin R.,2009,A consensus list of microsatellite markers for olive genotyping,Mol.Breeding,24(3):213-231
Carriero F.,Fontanazza G.,Cellini F.,and Giorio G.,2002,Identification of simple sequence repeats(SSRs)in olive(Olea europaea L),Theor.Appl.Genet.,104(2-3):301-307
Cipriani G.,Marazzo M.T.,Marconi R.,Cimato A.,and Testolin R.,2002,Microsatellite markers isolated in olive(Olea europaea L.)are suitable fo r individual fingerprinting and reveal polymorphism within ancient cultivars,Theor.Appl.Genet.,104(2-3):223-228
Collani S.,Galla G.,Ramina A.,Barcaccia G.,Alagna F.,Caceres E.M.,Baldoni L.,Muleo R.,and Perrotta G.,2012,Self-incompatibility in olive:a new hypothesis on the S-locus genes controlling pollen-pistil interaction,Acta Hortic.,967(1):133-140
Díaz A.,Martín A.,Rallo P.,and Rosa R.D.L.,2006a,Self-incompatibility of'Arbequina'and'Picual'olive assessed by SSR markers,J.Amer.Soc.Hort.Sci.,131(2):250-255
Díaz A.,Rosa R.D.L.,Martín A.,and Rallo P.,2006b,Development,characterization and inheritance of new microsatellites in olive(Olea europaea L.)and evaluation of their usefulness in cultivar identification and genetic relationship studies,Tree Genet.Genomes,2(3):165-175
Díaz A.,Martín A.,Rallo P.,and Rosa R.D.L.,2007,Cross-compatibility of the parents as the main factor for successful olive breeding crosses,J.Amer.Soc.Hort.Sci.,132(6):830-835
Doveri S.,Gil S.F.,Díaz A.,Reale S.,Busconi M.,Machado A.C.,Martin A.,Fogher C.,Donini P.,and Lee D.,2008,Standardization of a set of microsatellite markers for use in cultivar identification studies in olive(Olea europaea L.),Sci.Hortic.,116(4):367-373
Kapellakis I.E.,Tsagarakis K.P.,and Crowther J.C.,2008,Olive oil history,production and by-product management,Rev.Environ.Sci.Biotechno.,7(1):1-26
Ma T.,Ning D.L.,Yang W.M.,Zhang Z.Z.,Li Y.J.,Xu T.,and Xiao L.J.,2014,Breeding of a new olive cultivar'Jinyefoxi lan',Zhongguo Guoshu(China Fruits),(6):3-4,85(马婷,宁德鲁,杨卫明,张植中,李勇杰,徐田,肖良俊,2014,油橄榄新品种金叶佛樨榄的选育,中国果树,(6):3-4,85)
Ma T.,Xu T.,Ning D.L.,Xiao L.J.,and Li J.,2015,Comparative Study on the growth and morphology of new olive varieties'Jinyefoxilan'and its parents,Nanfang Yuanyi(Southern hor-ticulture),26(4):1-3(马婷,徐田,宁德鲁,肖良俊,李静,2015,油橄榄新品种“金叶佛樨榄”与其父母本生长与形态的比较研究,南方园艺,26(4):1-3)
Mookerjee S.,Guerin J.,Collins G.,Ford C.,and Sedgley M.,2005,Paternity analysis using microsatellite markers to identify pollen donors in an olive grove,Theor.Appl.Genet.,111(6):1174-1182
Rosa R.D.L.,James C.M.,and Tobutt K.R.,2002,Isolation and characterization of polymorphic microsatellites in olive(Olea europaea L.)and their transferability to other genera in the Oleaceae,Mol.Ecol.Notes,2(3):265-267
Rosa R.D.L.,James C.M.,and Tobutt K.R.,2004,Using microsatellites for paternity testing in olive progenies,Hort.Sci.,39(2):351-354
Sefc K.M.,Lopes M.S.,Mendonca D.,Santos M.R.D.,and Machado A.D.C.,2010,Identification of microsatellite loci in olive(Olea europaea L.)and their characterization in Italian and Iberian olive trees,Mol.Ecol.,9(8):1171-1173
Wang J.N.,Zhou M.M.,Shang S.,Tian L.B.,and Pan Q.Y.,2018,Optimization of bitter gourd SSR-PCR reaction system and shuttle of its primer in the melon crops,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),16(6):1869-1877(王佳楠,周萌萌,商桑,田丽波,潘琼玉,2018,苦瓜SSR-PCR体系优化及其引物在瓜类作物中的穿梭性,分子植物育种,16(6):1869-1877)
Wang R.J.,and Li P.Y.,2016,Analysis on transferability of wheat EST-SSR markers in Elytrigia repens,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(6):1516-1523(王瑞晶,李培英,2016,小麦EST-SSR标记在偃麦草中的通用性分析,分子植物育种,14(6):1516-1523)
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)
Zhang Y.Y.,Qi H.,Guo Q.M.,Sun Z.Y.,Zhou F.Q.,and Li S.B.,2016,Analysis of genetic diversity in Chaenomeles using apple EST-SSRs,Shengwu Jishu Tongbao(Biotechnology Bulletin),32(7):93-98(张艳艳,齐红,郭庆梅,孙稚颖,周凤琴,李圣波,2016,利用苹果EST-SSR分析木瓜属种质遗传多样性,生物技术通报,32(7):93-98)
Carriero F.,Fontanazza G.,Cellini F.,and Giorio G.,2002,Identification of simple sequence repeats(SSRs)in olive(Olea europaea L),Theor.Appl.Genet.,104(2-3):301-307
Cipriani G.,Marazzo M.T.,Marconi R.,Cimato A.,and Testolin R.,2002,Microsatellite markers isolated in olive(Olea europaea L.)are suitable fo r individual fingerprinting and reveal polymorphism within ancient cultivars,Theor.Appl.Genet.,104(2-3):223-228
Collani S.,Galla G.,Ramina A.,Barcaccia G.,Alagna F.,Caceres E.M.,Baldoni L.,Muleo R.,and Perrotta G.,2012,Self-incompatibility in olive:a new hypothesis on the S-locus genes controlling pollen-pistil interaction,Acta Hortic.,967(1):133-140
Díaz A.,Martín A.,Rallo P.,and Rosa R.D.L.,2006a,Self-incompatibility of'Arbequina'and'Picual'olive assessed by SSR markers,J.Amer.Soc.Hort.Sci.,131(2):250-255
Díaz A.,Rosa R.D.L.,Martín A.,and Rallo P.,2006b,Development,characterization and inheritance of new microsatellites in olive(Olea europaea L.)and evaluation of their usefulness in cultivar identification and genetic relationship studies,Tree Genet.Genomes,2(3):165-175
Díaz A.,Martín A.,Rallo P.,and Rosa R.D.L.,2007,Cross-compatibility of the parents as the main factor for successful olive breeding crosses,J.Amer.Soc.Hort.Sci.,132(6):830-835
Doveri S.,Gil S.F.,Díaz A.,Reale S.,Busconi M.,Machado A.C.,Martin A.,Fogher C.,Donini P.,and Lee D.,2008,Standardization of a set of microsatellite markers for use in cultivar identification studies in olive(Olea europaea L.),Sci.Hortic.,116(4):367-373
Kapellakis I.E.,Tsagarakis K.P.,and Crowther J.C.,2008,Olive oil history,production and by-product management,Rev.Environ.Sci.Biotechno.,7(1):1-26
Ma T.,Ning D.L.,Yang W.M.,Zhang Z.Z.,Li Y.J.,Xu T.,and Xiao L.J.,2014,Breeding of a new olive cultivar'Jinyefoxi lan',Zhongguo Guoshu(China Fruits),(6):3-4,85(马婷,宁德鲁,杨卫明,张植中,李勇杰,徐田,肖良俊,2014,油橄榄新品种金叶佛樨榄的选育,中国果树,(6):3-4,85)
Ma T.,Xu T.,Ning D.L.,Xiao L.J.,and Li J.,2015,Comparative Study on the growth and morphology of new olive varieties'Jinyefoxilan'and its parents,Nanfang Yuanyi(Southern hor-ticulture),26(4):1-3(马婷,徐田,宁德鲁,肖良俊,李静,2015,油橄榄新品种“金叶佛樨榄”与其父母本生长与形态的比较研究,南方园艺,26(4):1-3)
Mookerjee S.,Guerin J.,Collins G.,Ford C.,and Sedgley M.,2005,Paternity analysis using microsatellite markers to identify pollen donors in an olive grove,Theor.Appl.Genet.,111(6):1174-1182
Rosa R.D.L.,James C.M.,and Tobutt K.R.,2002,Isolation and characterization of polymorphic microsatellites in olive(Olea europaea L.)and their transferability to other genera in the Oleaceae,Mol.Ecol.Notes,2(3):265-267
Rosa R.D.L.,James C.M.,and Tobutt K.R.,2004,Using microsatellites for paternity testing in olive progenies,Hort.Sci.,39(2):351-354
Sefc K.M.,Lopes M.S.,Mendonca D.,Santos M.R.D.,and Machado A.D.C.,2010,Identification of microsatellite loci in olive(Olea europaea L.)and their characterization in Italian and Iberian olive trees,Mol.Ecol.,9(8):1171-1173
Wang J.N.,Zhou M.M.,Shang S.,Tian L.B.,and Pan Q.Y.,2018,Optimization of bitter gourd SSR-PCR reaction system and shuttle of its primer in the melon crops,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),16(6):1869-1877(王佳楠,周萌萌,商桑,田丽波,潘琼玉,2018,苦瓜SSR-PCR体系优化及其引物在瓜类作物中的穿梭性,分子植物育种,16(6):1869-1877)
Wang R.J.,and Li P.Y.,2016,Analysis on transferability of wheat EST-SSR markers in Elytrigia repens,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(6):1516-1523(王瑞晶,李培英,2016,小麦EST-SSR标记在偃麦草中的通用性分析,分子植物育种,14(6):1516-1523)
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)
Zhang Y.Y.,Qi H.,Guo Q.M.,Sun Z.Y.,Zhou F.Q.,and Li S.B.,2016,Analysis of genetic diversity in Chaenomeles using apple EST-SSRs,Shengwu Jishu Tongbao(Biotechnology Bulletin),32(7):93-98(张艳艳,齐红,郭庆梅,孙稚颖,周凤琴,李圣波,2016,利用苹果EST-SSR分析木瓜属种质遗传多样性,生物技术通报,32(7):93-98)