甜瓜单性雌花共显性分子标记的开发及应用
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摘要
单性雌花是甜瓜育种中的重要目标性状之一,其在种子生产过程中可提高纯度,降低人工成本。当前控制甜瓜单性雌花基因CmACS-7已经被克隆并完成了功能验证。本研究根据甜瓜纯合单性雌花材料与两性花材料在CmACS-7基因缺失位点(Insert/Deletion),开发出了InDel-1标记。利用该分子标记对25份甜瓜材料进行基因型检测。结果显示:甜瓜单性雌花材料表现为缺失带型或杂合带型,两性花则表现为非缺失带型,标记多态性与花性型共分离。此外利用InDel分子标记辅助育种,对四个由纯合单性雌花与两性花杂交得到的F2代分离群体,进行花性型的鉴定,检测准确率都达到94.0%以上。研究表明:InDel-1标记在甜瓜单性雌花的实际鉴定中具有较高的准确性,能够大大提高品种选育的效率,缩短育种周期。
Monoecious is one of the important target traits in melon breeding, which could improve the purity of seeds and reduced the labor cost in seed production. The monoecious gene CmACS-7 has been cloned and functionally verified in melon. In this study, the InDel-1 marker was developed based on homozygous monoecious and andromonoecious at the CmACS-7 gene deletion site(Insert/Deletion). 25 melon materials were genotyped using this marker, and results showed that the monoecious showed a missing band or a heterozygous band. The andromonoecious showed no missing bands, and marked the co-segregated of marker polymorphism and flower sexual type. Four F2 populations segregating generations were constructed from the hybrid combinations with homozygous monoecious and andromonoecious by InDel molecular marker-assisted breeding, and identified the flower sexual type. The accuracy of genotyping results reached over 94.0% respectively in four populations.The results showed that Indel-1 marker might have high accuracy, which could abundantly raise the efficiency of breeding and shorten the breeding cycle.
Monoecious is one of the important target traits in melon breeding, which could improve the purity of seeds and reduced the labor cost in seed production. The monoecious gene CmACS-7 has been cloned and functionally verified in melon. In this study, the InDel-1 marker was developed based on homozygous monoecious and andromonoecious at the CmACS-7 gene deletion site(Insert/Deletion). 25 melon materials were genotyped using this marker, and results showed that the monoecious showed a missing band or a heterozygous band. The andromonoecious showed no missing bands, and marked the co-segregated of marker polymorphism and flower sexual type. Four F2 populations segregating generations were constructed from the hybrid combinations with homozygous monoecious and andromonoecious by InDel molecular marker-assisted breeding, and identified the flower sexual type. The accuracy of genotyping results reached over 94.0% respectively in four populations.The results showed that Indel-1 marker might have high accuracy, which could abundantly raise the efficiency of breeding and shorten the breeding cycle.
引文
Bhowmick B.K.,and Jh a S.,2015,Dynamics of sex expression and chromosome diversity in Cucurbitaceae:a story in the making,J.Genet.,94(4):793-808
Boualem A.,Troadec C.,Kovalski I.,Sari M.A.,Perl-Treves R.,and Bendahmane A.,2009,A conserved ethylene biosynthesis enzyme leads to andromonoecy in two cucumis species,PLoS One,4(7):e6144
Carvalho N.,Canela F.M.,Leite P.,Ferreira M.A.,Oliveira V.R.,Santos M.F.,Souza N.,and Buso G.,2017,Analysis of genetic variability of commercial melon cultivars using SSRmolecular markers,Genet.Mol.Res.,16(3):1-8
Collard B.C.Y.,and Mackill D.J.,2008,Marker-assisted selection:an approach for precision plant breeding in the twenty-first century,Philos.Trans.R Soc.Lond B Biol.Sci.,363(1491):557-572
Feng H.,Li X.M.,Liu Z.Y,Wei P.,and Ji R.Q.,2009,A co-dominant molecular marker linked to the monoecious gene CmACS-7 derived from gene sequence in Cucumis melo L.,African Journal of Biotechnology,8(14):3168-3174
Fernandoj Y.L.,Carmen C.,Emilio S.,Rocio T.,Marial G.G.,Juan C.,Rafael L.,and Ana I.L.,2011,Genetic linkage map of melon(Cucumis melo L.)and localization of a major QTLfor powdery mildew resistance,Mol.Breeding,27(2):181-192
Gong H.,Luo J.N.,Luo S.B.,Zheng X.M.,He X.L.,and Wu H.B.,2014,Progress of study on the molecular mechanism of sex-determination in melon and cucumber,Yuanyi Xuebao(Acta Horticulturae Sinica),41(2):382-388(龚浩,罗剑宁,罗少波,郑晓明,何晓莉,吴海滨,2014,黄瓜与甜瓜的性别决定分子机制研究进展,园艺学报,41(2):382-388)
Li B.,Zhao Y.,Zhu Q.,Zhang Z.,Fan C.,Amanullah S.,Gao P.,and Luan F.,2017,Mapping of powdery mildew resistance genes in melon(Cucumis melo L.)by bulked segregant analysis,Sci.Hortic.,220(16):160-167
Liang L.,Li R.F.,Li K.,Du G.F.,and Li Y.L.,2003,Studies on transferring the character of single-sex flower in oriental melon,Huabei Nongxuebao(North China Agriculture Re port),18(2):78-80(梁莉,李荣富,李堃,都桂芳,李玉莲,2003,薄皮甜瓜单性花性状转育研究,华北农学报,18(2):78-80)
Martin A.,Troadec C.,Boualem A.,Rajab M.,Fernandez R.,Morin H.,Pitrat M.,Dogimont C.,and Bendahmane A.,2009,A transposon-induced epigenetic change leads to sex determination in melon,Nature,461(7267):1135-1138
Murray M.G.,and Thompson W.F.,1980,Rapid isolation of high molecular weight plant DNA,Nucleic Acids Res.,8(19):4321-4325
Song X.,Wei H.,Cheng W.,Yang S.,Zhao Y.,Li X.,Luo D.,Zhang H.,and Feng X.,2015,Development of INEL markers for genetic mapping based on whole genome resequencing in soybean,G3(Bethesda,Md.),5(12):2793-2799
Sousaraei N.,Ramshini H.,Lotfi M.,and Sharzei A.,2018,Marker assisted backcrossing for introgression of Fusarium wilt resistance gene into melon,Euphytica,214(1):7
Tzuri G.,Zhou X.,Chayut N.,Yuan H.,Portnoy V.,Meir A.,Sa'Ar U.,Baumkoler F.,Mazourek M.,Lewinsohn E.,Fei Z.,Schaffer A.A.,Li L.,Burger J.,Katzir N.,and Tadmor Y.A.,2015,'Golden'SNP in Cm Or governs the fruit flesh color of melon(Cucumis melo),Plant J.,82(2):267-279
Yang J.,He J.,Wang D.B.,Shi E.,Yang W.Y.,Geng Q.F.,and Wang Z.S.,2016,Progress in research and application of InDel markers,Shengwu Duoyangxing(Biodiversity Science),24(2):237-243(杨洁,赫佳,王丹碧,施恩,杨文宇,耿其芳,王中生,2016,InDel标记的研究和应用进展,生物多样性,24(2):237-243)
Yang X.H.,Hua D.P.,Fu J.Y.,Zhang Q.L.,Xie M.T.,and Liu L.,2018,Development and application of a molecular for melon flesh color,Xibei Zhiwu Xuebao(Acta Botanica BorealiOccidentalia Sinica),38(4):654-660(杨旭辉,华德平,付金玉,张乔玲,谢梅婷,刘莉,2018,甜瓜果肉颜色相关的分子标记开发和应用,西北植物学报,38(4):654-660)
Yu R.,Zhang X.,Yu Y.,Yu Y.M.,and Yang J.Q.,2006,Studies on selective breeding of monoecious muskmelon and analysis of heredity regular,Xibei Nonglin Keji Daxue Xuebao(Journal of Northwest A&F University(Natural Science Edition)),34(11):160-162(于蓉,张显,于远,杨玉梅,杨建强,2006,厚皮甜瓜单性花系的选育及其遗传分析,西北农林科技大学学报(自然科学版),34(11):160-162)
Zhang H.J.,Li Y.,Yu J.,and Dai Y.,2016,Research progress in gene engineering of melon,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(3):608-612(张慧君,李烨,余静,戴誉,2016,甜瓜基因工程研究进展,分子植物育种,14(3):608-612)
Boualem A.,Troadec C.,Kovalski I.,Sari M.A.,Perl-Treves R.,and Bendahmane A.,2009,A conserved ethylene biosynthesis enzyme leads to andromonoecy in two cucumis species,PLoS One,4(7):e6144
Carvalho N.,Canela F.M.,Leite P.,Ferreira M.A.,Oliveira V.R.,Santos M.F.,Souza N.,and Buso G.,2017,Analysis of genetic variability of commercial melon cultivars using SSRmolecular markers,Genet.Mol.Res.,16(3):1-8
Collard B.C.Y.,and Mackill D.J.,2008,Marker-assisted selection:an approach for precision plant breeding in the twenty-first century,Philos.Trans.R Soc.Lond B Biol.Sci.,363(1491):557-572
Feng H.,Li X.M.,Liu Z.Y,Wei P.,and Ji R.Q.,2009,A co-dominant molecular marker linked to the monoecious gene CmACS-7 derived from gene sequence in Cucumis melo L.,African Journal of Biotechnology,8(14):3168-3174
Fernandoj Y.L.,Carmen C.,Emilio S.,Rocio T.,Marial G.G.,Juan C.,Rafael L.,and Ana I.L.,2011,Genetic linkage map of melon(Cucumis melo L.)and localization of a major QTLfor powdery mildew resistance,Mol.Breeding,27(2):181-192
Gong H.,Luo J.N.,Luo S.B.,Zheng X.M.,He X.L.,and Wu H.B.,2014,Progress of study on the molecular mechanism of sex-determination in melon and cucumber,Yuanyi Xuebao(Acta Horticulturae Sinica),41(2):382-388(龚浩,罗剑宁,罗少波,郑晓明,何晓莉,吴海滨,2014,黄瓜与甜瓜的性别决定分子机制研究进展,园艺学报,41(2):382-388)
Li B.,Zhao Y.,Zhu Q.,Zhang Z.,Fan C.,Amanullah S.,Gao P.,and Luan F.,2017,Mapping of powdery mildew resistance genes in melon(Cucumis melo L.)by bulked segregant analysis,Sci.Hortic.,220(16):160-167
Liang L.,Li R.F.,Li K.,Du G.F.,and Li Y.L.,2003,Studies on transferring the character of single-sex flower in oriental melon,Huabei Nongxuebao(North China Agriculture Re port),18(2):78-80(梁莉,李荣富,李堃,都桂芳,李玉莲,2003,薄皮甜瓜单性花性状转育研究,华北农学报,18(2):78-80)
Martin A.,Troadec C.,Boualem A.,Rajab M.,Fernandez R.,Morin H.,Pitrat M.,Dogimont C.,and Bendahmane A.,2009,A transposon-induced epigenetic change leads to sex determination in melon,Nature,461(7267):1135-1138
Murray M.G.,and Thompson W.F.,1980,Rapid isolation of high molecular weight plant DNA,Nucleic Acids Res.,8(19):4321-4325
Song X.,Wei H.,Cheng W.,Yang S.,Zhao Y.,Li X.,Luo D.,Zhang H.,and Feng X.,2015,Development of INEL markers for genetic mapping based on whole genome resequencing in soybean,G3(Bethesda,Md.),5(12):2793-2799
Sousaraei N.,Ramshini H.,Lotfi M.,and Sharzei A.,2018,Marker assisted backcrossing for introgression of Fusarium wilt resistance gene into melon,Euphytica,214(1):7
Tzuri G.,Zhou X.,Chayut N.,Yuan H.,Portnoy V.,Meir A.,Sa'Ar U.,Baumkoler F.,Mazourek M.,Lewinsohn E.,Fei Z.,Schaffer A.A.,Li L.,Burger J.,Katzir N.,and Tadmor Y.A.,2015,'Golden'SNP in Cm Or governs the fruit flesh color of melon(Cucumis melo),Plant J.,82(2):267-279
Yang J.,He J.,Wang D.B.,Shi E.,Yang W.Y.,Geng Q.F.,and Wang Z.S.,2016,Progress in research and application of InDel markers,Shengwu Duoyangxing(Biodiversity Science),24(2):237-243(杨洁,赫佳,王丹碧,施恩,杨文宇,耿其芳,王中生,2016,InDel标记的研究和应用进展,生物多样性,24(2):237-243)
Yang X.H.,Hua D.P.,Fu J.Y.,Zhang Q.L.,Xie M.T.,and Liu L.,2018,Development and application of a molecular for melon flesh color,Xibei Zhiwu Xuebao(Acta Botanica BorealiOccidentalia Sinica),38(4):654-660(杨旭辉,华德平,付金玉,张乔玲,谢梅婷,刘莉,2018,甜瓜果肉颜色相关的分子标记开发和应用,西北植物学报,38(4):654-660)
Yu R.,Zhang X.,Yu Y.,Yu Y.M.,and Yang J.Q.,2006,Studies on selective breeding of monoecious muskmelon and analysis of heredity regular,Xibei Nonglin Keji Daxue Xuebao(Journal of Northwest A&F University(Natural Science Edition)),34(11):160-162(于蓉,张显,于远,杨玉梅,杨建强,2006,厚皮甜瓜单性花系的选育及其遗传分析,西北农林科技大学学报(自然科学版),34(11):160-162)
Zhang H.J.,Li Y.,Yu J.,and Dai Y.,2016,Research progress in gene engineering of melon,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(3):608-612(张慧君,李烨,余静,戴誉,2016,甜瓜基因工程研究进展,分子植物育种,14(3):608-612)