白菜型油菜黄籽沙逊和芥蓝种间杂交合成甘蓝型油菜研究
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摘要
人工合成甘蓝型油菜是拓宽油菜遗传育种种质资源的有效途径,也是研究芸薹属物种起源与演化的重要手段之一。为了把黄籽沙逊的黄籽性状同芥蓝的优良特性相结合,本研究利用白菜型油菜黄籽沙逊(Brassica. campestris var. Yellow Sarson Pain, 2n=AA=20)与芥蓝(Brassica albograbra, 2n=CC=18)开展了芸薹属种间杂交研究,在MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+7%蔗糖+活性炭1.0 mg/L的培养基上,通过胚珠培养从授粉7 d的140个胚珠中得到6个远缘杂交胚发育而成的植株,其中1个自然加倍成为双二倍体,随后应用形态学、细胞学和分子标记的方法对双二倍体种间杂种植株进行了鉴定,结果表明杂种植株生长势较强,形态属于中间类型;根尖染色体压片观察显示杂种植株具有父母本的染色体数量之和;SSR分子标记分析表明,杂种植株包含了双亲的遗传信息,由此证明了所获杂种的真实性,这为最后自交分离获得黄籽甘蓝型油菜提供了科学依据。
Synthetic Brassica napus is an effective way to broaden the germplasm resources of rape breeding and an important way to study the origin and evolution of Brassica species. In order to combine the yellow seed of'Yellow Sarson' with the fine traits of B. albograbra, in this research, the intergeneric hybridization experiment between 'Yellow Sarson'(Brassica campestris, AA genome, 2 n=20) and B. albograbra(CC genome, 2 n=18) were car ried out. 6 intergeneric hybrids were obtained from 140 ovules of pollinated for 7 d on the medium(MS+6-BA1.0 mg/L+NAA 0.1 mg/L+7% sucrose+carbon 1.0 mg/L) through embryo rescue. One of 6 embryos became double diploid through natural chromosome doubling, which was subsequently identified by morphological characteriza-tion, cytological observation and molecular marker techniques. The results indicated that the hybrids showed stronger growth vigor compared with their parents and belonged to middle type in morphology. Cytological observation by slide preparation of chromosomes from root tip cells revealed that the hybrid plants contained the all chromosome of both parents. SSR molecular marker analysis showed that the hybrids contained genetic information of both parents, which proved that the hybrids were real hybrids. The creation and identification of the hybrids could lay the basis for the obtain of yellow-seed Brassica napus by selfing.
Synthetic Brassica napus is an effective way to broaden the germplasm resources of rape breeding and an important way to study the origin and evolution of Brassica species. In order to combine the yellow seed of'Yellow Sarson' with the fine traits of B. albograbra, in this research, the intergeneric hybridization experiment between 'Yellow Sarson'(Brassica campestris, AA genome, 2 n=20) and B. albograbra(CC genome, 2 n=18) were car ried out. 6 intergeneric hybrids were obtained from 140 ovules of pollinated for 7 d on the medium(MS+6-BA1.0 mg/L+NAA 0.1 mg/L+7% sucrose+carbon 1.0 mg/L) through embryo rescue. One of 6 embryos became double diploid through natural chromosome doubling, which was subsequently identified by morphological characteriza-tion, cytological observation and molecular marker techniques. The results indicated that the hybrids showed stronger growth vigor compared with their parents and belonged to middle type in morphology. Cytological observation by slide preparation of chromosomes from root tip cells revealed that the hybrid plants contained the all chromosome of both parents. SSR molecular marker analysis showed that the hybrids contained genetic information of both parents, which proved that the hybrids were real hybrids. The creation and identification of the hybrids could lay the basis for the obtain of yellow-seed Brassica napus by selfing.
引文
Cheng F.,Liu S.Y,Wu J.,Fang L.,Sun S.L,Liu B.,Li P.X,Hua W.,and Wang X.,2011,BRAD,the genetics and genomics database for Brassica plants,BMC Plant Biol.,11:136
Daun J.K.,and DeClercq D.R.,1988,Quality of yellow and dark seeds in Brassica campestris canola varieties Candle and Tobin,J.Am.Oil Chem.Soc.,65(1):122-126
Du P.,Zhang X.Y.,Li L.N.,Huang B.Y.,Yi M.L.,Dong W.Z.and Tang F.S.,2013,Study on slide preparation methods for high quality chromosomes from root tip cells of Arechis Henan Nongye Kexue(Journal of Henan Agricultural Sciences),42(3):31-35(杜培,张新友,李丽娜,黄冰艳,易明林,董文召,汤丰收,2013,高质量花生根尖细胞染色体制片方法研究,河南农业科学,42(3):31-35)
Feng W.,Deng Y.F.,Li M.X.,and Zhang X.Y.,1993,The cytoplasmic inheritance displayed in the reciprocal crosses between B.pekinensis and B.oleracea and its prospect in utilization,Yuanyi Xuebao(Acta Horticulturae Sinica),20(3):267-273(冯午,邓岳芬,李懋学,张新英,1993,白菜甘蓝种间杂种后代细胞质遗传及其利用前景探讨,园艺学报,20(3):267-273)
Fu S.H.,Li Y.,Yang J.,Wang J.S.,Chen X.H.,Zou Q.T.,Lan R.Kang Z.M.,Tang R.,and Zhang R.Q.,2012,Study on obtain ing of hybrids(B.napus×B.campestris)by chromosome doubling method,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),25(4):1164-1170(付绍红,李云,杨进,王继胜,陈晓华,邹琼陶,兰蓉,康泽明,唐蓉,张汝全,2012,染色体加倍获得甘白(甘蓝型×白菜型油菜)杂交后代研究,西南农业学报,25(4):1164-1170)
Ge K.L.,1964,Synthesis of new species and cytogenetic studies in oleiferous BrassicaⅠ:studies of an artificially synthesized Napus(Brassica napus),Fudan Daxue Xuebao(Fudan University Journal(Natural Science Edition)),9(3):393-412(葛扣麟,1964,芸薹属油菜植物的新种合成及其细胞遗传学研究,一个人工合成欧洲油菜(B.napus)的研究,复旦大学学报(自然科学版),9(3):393-412)
He Y.T.,Chen B.Y.,Fu T.D.,Li D.R.,and Tu J.X.,2003,Origins and evolution of Brassica campestris L.in China,Yichuan Xuebao(Acta Genetica Sinica),30(11):1003-1012(何余堂,陈宝元,傅廷栋,李殿荣,涂金星,2003,白菜型油菜在中国的起源与进化,遗传学报,30(11):1003-1012)
Hu X.M.,Li X.X.,Liang Y.,Fang Z.Y.,Shen D.,and Qiu Y.,2009,Optimization of technology for embryo rescue in distant hybridization between cabbage and radish,Zhongguo Shucai(China Vegetables),(10):7-12(扈新民,李锡香,梁燕,方智远,沈镝,邱杨,2009,甘蓝与萝卜属间杂交及其胚挽救技术优化,中国蔬菜,(10):7-12)
Inomata N.,1977,Production of inter specific hybrids between B.campestris and B.oleracea by culture in vitro of excised ovaries:Ⅰ.Effects of yeast extract and case in hydrolysate on the development of excised ovaries,Japan J.Breed.,27(4):295-304
Inomata N.,1979,Production of inter specific hybrids between B.campestris and B.oleracea by culture in vitro of excised ovaries:Ⅱ.development of excised ovaries on various culture media,Japan J.Breed.,29(2):115-120
Inomata N.,1980,Hybrid progenies of the cross between B.campestris and B.oleracea,Japan J.Breed.,55:189-202
Liang H.,and Feng W.,1990,The interspecific cross-Brassica pekinensis×B.oleracea and observations on the F1proge nies,Yuanyi Xuebao(Acta Horticulture Sinica),17(3):203-210(梁红,冯午,1990,白菜与甘蓝的种间杂交及其后代的研究,园艺学报,17(3):203-210)
Momotaz A.,Kato M.,and Kakihara F.,1998,Production of intergeneric hybridization between Brassica and Sinapis species by means of embryo rescue techniques,Euphytica,103:123-130
Nagaharu U.,1935,Genome analysis in Brassica with special reference to the experimental formation of B.napus and peculiar mode of fertilization,Jan.J.Bot.,7(7):389-452
Quazi M.H.,1988,Interspecific hybrids between B.napus L.,and B.oleracea L.,developed by embryo culture,Theor.Appl.Genet.,75(2):309-318
Rahman M.H.,2001,Production of yellow-seeded Brassica napus through interspecific crosses,Plant Breed.,120(6):463-472
Rahman M.H.,Joersbo M.,and Poulsen M.H.,2001,Development of yellow-seeded Brassica napus of double low quality,Plant Breed.,120(6):473-478
Shirzadegan M.,and Robbelen G.,1985,Influence of seed colour and hull proportions on quality properties of seeds in Brassica napus L.,Fette.Seifen.Anstrichm.,87(6):235-237
Slominski B.A.,Campbell L.D.,and Guenter W.,1994,Carbohydrates and dietary fibre components of yellow and brownseeded canola,J.Agric.Food Chem.,42(3):704-707
Tan Y.F.,Zhang Q.,Zhao Y.Y.,Xu Q.,Li Y.,Yuan Y.X.,Yao Q.J.,Jiang W.S.,Wei X.C.,Wang Z.Y.,and Zhang X.W.2017,Identification of interspecific hybrid between Yellow Sarson and yellow seeded Ethiopianmustard,Henan Nongye Kexue(Journal of Henan Agricultural Sciences),46(3):107-111(谭亚飞,张强,赵艳艳,许茜,李扬,原玉香,姚秋菊,蒋武生,魏小春,王志勇,张晓伟,2017,黄籽沙逊与黄籽埃塞俄比亚芥种间杂种的鉴定,河南农业科学,46(3):107-111)
Tang Z.L.,Li J.N.,Zhang X.K.,Chen L.,and Wang R.,1997,Genetic variation of yellow-seeded rapeseed lines(Brassica napus L.)from different genetic sources,Plant Breed.,116(5)471-474
Wen Y.C.,Zhang S.F.,Wang J.P.,Li G.Y.,and Zhang X.W.1999,Studies on synthetic napus by interspecific bybridization between B.ocleracea and B.campestris,Zhongguo Youliao Zuowu Xuebao(Chinese Journal of Oil Crops Sciences),21(4):8-11(文雁成,张书芬,王建平,栗根义,张晓伟,1999,对甘蓝与大白菜种间杂交合成的甘蓝型油菜的研究,中国油料作物学报,21(4):8-11)
Whetten R.W.,Mackay J.J.,and Sederoff R.R.,1998,Recent advances in understanding lignin biosynthesis,Annu.Rev Plant Physiol.Mol.Biol.,49(1):585-609
Yin J.M.,Chen S.Z.,Tang Z.L.,Chen L.,and Li J.N.,2004Resynthesis of Brassica napus through interspecific hybridization between yellow-seeded B.oleracea var.acephala and B.campestris,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),17(2):149-151(殷家明陈树忠,唐章林,谌利,李加纳,2004,黄籽羽衣甘蓝和白菜型油菜杂交再合成甘蓝型油菜研究,西南农业学报,17(2):149-151)
Zhang R.M.,and An X.Z.,2006,Research progress of genetic and breeding on yellow seed variety of Brassica napus,Guizhou Nongye Kexue(Guizhou Agricultural Sciences),34(2):75-78(张瑞茂,安兴智,2006,甘蓝型黄籽油菜遗传及育种研究进展,贵州农业科学,34(2):75-78)
Zhao Y.Y.,Jiang W.S.,Yuan Y.X.,Wang Z.Y.,Yao Q.J.,Wei X.C.,Zhang Q.,and Zhang X.W.,2016,Embryo rescue of Brassica oleracea L.var.capitata×Brassica napus L.and their ploidy identification,Henan Nongye Kexue(Journal of Henan Agricultural Sciences),45(9):88-93(赵艳艳,蒋武生,原玉香,王志勇,姚秋菊,魏小春,张强,张晓伟,2016,结球甘蓝与油菜杂交胚挽救及其后代倍性鉴定,河南农业科学,45(9):88-93)
Zhou Q.Y.,Yin J.M.,Cui C.,Lin N.,Chen L.,Tang Z.L.,and Li J.N.,2006,Characters of interspecific hybrids between Brassica campestris and B.oleracea var.Acephala,Xinan Nongye Daxue Xuebao(Journal of Southwest Agricultural University),28(1):1-3(周清元,殷家明,崔翠,林呐,谌利,唐章林,李加纳,2006,白菜型油菜×羽衣甘蓝种间杂种的性状表现,西南农业大学学报,28(1):1-3)
Zhou Q.Y.,Li J.N.,Yin J.M.,Tang Z.L.,Chen L.,and Tao L.,2003,Preliminary studies on interspecific hybridization between Brassica campestris and B.oleracea AceaphalaⅠ:The effect of the time of overay taking on seed froming in vitro,Xinan Nongye Daxue Xuebao(Journal of Southwest Agricultural University),25(6):495-497(周清元,李加纳,殷家明,唐章林,谌利,陶澜,2003,白菜型油菜和羽衣甘蓝种间杂交的初步研究Ⅰ,取材时间对子房离体培养结籽率的影响,西南农业大学学报,25(6):495-497)
Daun J.K.,and DeClercq D.R.,1988,Quality of yellow and dark seeds in Brassica campestris canola varieties Candle and Tobin,J.Am.Oil Chem.Soc.,65(1):122-126
Du P.,Zhang X.Y.,Li L.N.,Huang B.Y.,Yi M.L.,Dong W.Z.and Tang F.S.,2013,Study on slide preparation methods for high quality chromosomes from root tip cells of Arechis Henan Nongye Kexue(Journal of Henan Agricultural Sciences),42(3):31-35(杜培,张新友,李丽娜,黄冰艳,易明林,董文召,汤丰收,2013,高质量花生根尖细胞染色体制片方法研究,河南农业科学,42(3):31-35)
Feng W.,Deng Y.F.,Li M.X.,and Zhang X.Y.,1993,The cytoplasmic inheritance displayed in the reciprocal crosses between B.pekinensis and B.oleracea and its prospect in utilization,Yuanyi Xuebao(Acta Horticulturae Sinica),20(3):267-273(冯午,邓岳芬,李懋学,张新英,1993,白菜甘蓝种间杂种后代细胞质遗传及其利用前景探讨,园艺学报,20(3):267-273)
Fu S.H.,Li Y.,Yang J.,Wang J.S.,Chen X.H.,Zou Q.T.,Lan R.Kang Z.M.,Tang R.,and Zhang R.Q.,2012,Study on obtain ing of hybrids(B.napus×B.campestris)by chromosome doubling method,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),25(4):1164-1170(付绍红,李云,杨进,王继胜,陈晓华,邹琼陶,兰蓉,康泽明,唐蓉,张汝全,2012,染色体加倍获得甘白(甘蓝型×白菜型油菜)杂交后代研究,西南农业学报,25(4):1164-1170)
Ge K.L.,1964,Synthesis of new species and cytogenetic studies in oleiferous BrassicaⅠ:studies of an artificially synthesized Napus(Brassica napus),Fudan Daxue Xuebao(Fudan University Journal(Natural Science Edition)),9(3):393-412(葛扣麟,1964,芸薹属油菜植物的新种合成及其细胞遗传学研究,一个人工合成欧洲油菜(B.napus)的研究,复旦大学学报(自然科学版),9(3):393-412)
He Y.T.,Chen B.Y.,Fu T.D.,Li D.R.,and Tu J.X.,2003,Origins and evolution of Brassica campestris L.in China,Yichuan Xuebao(Acta Genetica Sinica),30(11):1003-1012(何余堂,陈宝元,傅廷栋,李殿荣,涂金星,2003,白菜型油菜在中国的起源与进化,遗传学报,30(11):1003-1012)
Hu X.M.,Li X.X.,Liang Y.,Fang Z.Y.,Shen D.,and Qiu Y.,2009,Optimization of technology for embryo rescue in distant hybridization between cabbage and radish,Zhongguo Shucai(China Vegetables),(10):7-12(扈新民,李锡香,梁燕,方智远,沈镝,邱杨,2009,甘蓝与萝卜属间杂交及其胚挽救技术优化,中国蔬菜,(10):7-12)
Inomata N.,1977,Production of inter specific hybrids between B.campestris and B.oleracea by culture in vitro of excised ovaries:Ⅰ.Effects of yeast extract and case in hydrolysate on the development of excised ovaries,Japan J.Breed.,27(4):295-304
Inomata N.,1979,Production of inter specific hybrids between B.campestris and B.oleracea by culture in vitro of excised ovaries:Ⅱ.development of excised ovaries on various culture media,Japan J.Breed.,29(2):115-120
Inomata N.,1980,Hybrid progenies of the cross between B.campestris and B.oleracea,Japan J.Breed.,55:189-202
Liang H.,and Feng W.,1990,The interspecific cross-Brassica pekinensis×B.oleracea and observations on the F1proge nies,Yuanyi Xuebao(Acta Horticulture Sinica),17(3):203-210(梁红,冯午,1990,白菜与甘蓝的种间杂交及其后代的研究,园艺学报,17(3):203-210)
Momotaz A.,Kato M.,and Kakihara F.,1998,Production of intergeneric hybridization between Brassica and Sinapis species by means of embryo rescue techniques,Euphytica,103:123-130
Nagaharu U.,1935,Genome analysis in Brassica with special reference to the experimental formation of B.napus and peculiar mode of fertilization,Jan.J.Bot.,7(7):389-452
Quazi M.H.,1988,Interspecific hybrids between B.napus L.,and B.oleracea L.,developed by embryo culture,Theor.Appl.Genet.,75(2):309-318
Rahman M.H.,2001,Production of yellow-seeded Brassica napus through interspecific crosses,Plant Breed.,120(6):463-472
Rahman M.H.,Joersbo M.,and Poulsen M.H.,2001,Development of yellow-seeded Brassica napus of double low quality,Plant Breed.,120(6):473-478
Shirzadegan M.,and Robbelen G.,1985,Influence of seed colour and hull proportions on quality properties of seeds in Brassica napus L.,Fette.Seifen.Anstrichm.,87(6):235-237
Slominski B.A.,Campbell L.D.,and Guenter W.,1994,Carbohydrates and dietary fibre components of yellow and brownseeded canola,J.Agric.Food Chem.,42(3):704-707
Tan Y.F.,Zhang Q.,Zhao Y.Y.,Xu Q.,Li Y.,Yuan Y.X.,Yao Q.J.,Jiang W.S.,Wei X.C.,Wang Z.Y.,and Zhang X.W.2017,Identification of interspecific hybrid between Yellow Sarson and yellow seeded Ethiopianmustard,Henan Nongye Kexue(Journal of Henan Agricultural Sciences),46(3):107-111(谭亚飞,张强,赵艳艳,许茜,李扬,原玉香,姚秋菊,蒋武生,魏小春,王志勇,张晓伟,2017,黄籽沙逊与黄籽埃塞俄比亚芥种间杂种的鉴定,河南农业科学,46(3):107-111)
Tang Z.L.,Li J.N.,Zhang X.K.,Chen L.,and Wang R.,1997,Genetic variation of yellow-seeded rapeseed lines(Brassica napus L.)from different genetic sources,Plant Breed.,116(5)471-474
Wen Y.C.,Zhang S.F.,Wang J.P.,Li G.Y.,and Zhang X.W.1999,Studies on synthetic napus by interspecific bybridization between B.ocleracea and B.campestris,Zhongguo Youliao Zuowu Xuebao(Chinese Journal of Oil Crops Sciences),21(4):8-11(文雁成,张书芬,王建平,栗根义,张晓伟,1999,对甘蓝与大白菜种间杂交合成的甘蓝型油菜的研究,中国油料作物学报,21(4):8-11)
Whetten R.W.,Mackay J.J.,and Sederoff R.R.,1998,Recent advances in understanding lignin biosynthesis,Annu.Rev Plant Physiol.Mol.Biol.,49(1):585-609
Yin J.M.,Chen S.Z.,Tang Z.L.,Chen L.,and Li J.N.,2004Resynthesis of Brassica napus through interspecific hybridization between yellow-seeded B.oleracea var.acephala and B.campestris,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),17(2):149-151(殷家明陈树忠,唐章林,谌利,李加纳,2004,黄籽羽衣甘蓝和白菜型油菜杂交再合成甘蓝型油菜研究,西南农业学报,17(2):149-151)
Zhang R.M.,and An X.Z.,2006,Research progress of genetic and breeding on yellow seed variety of Brassica napus,Guizhou Nongye Kexue(Guizhou Agricultural Sciences),34(2):75-78(张瑞茂,安兴智,2006,甘蓝型黄籽油菜遗传及育种研究进展,贵州农业科学,34(2):75-78)
Zhao Y.Y.,Jiang W.S.,Yuan Y.X.,Wang Z.Y.,Yao Q.J.,Wei X.C.,Zhang Q.,and Zhang X.W.,2016,Embryo rescue of Brassica oleracea L.var.capitata×Brassica napus L.and their ploidy identification,Henan Nongye Kexue(Journal of Henan Agricultural Sciences),45(9):88-93(赵艳艳,蒋武生,原玉香,王志勇,姚秋菊,魏小春,张强,张晓伟,2016,结球甘蓝与油菜杂交胚挽救及其后代倍性鉴定,河南农业科学,45(9):88-93)
Zhou Q.Y.,Yin J.M.,Cui C.,Lin N.,Chen L.,Tang Z.L.,and Li J.N.,2006,Characters of interspecific hybrids between Brassica campestris and B.oleracea var.Acephala,Xinan Nongye Daxue Xuebao(Journal of Southwest Agricultural University),28(1):1-3(周清元,殷家明,崔翠,林呐,谌利,唐章林,李加纳,2006,白菜型油菜×羽衣甘蓝种间杂种的性状表现,西南农业大学学报,28(1):1-3)
Zhou Q.Y.,Li J.N.,Yin J.M.,Tang Z.L.,Chen L.,and Tao L.,2003,Preliminary studies on interspecific hybridization between Brassica campestris and B.oleracea AceaphalaⅠ:The effect of the time of overay taking on seed froming in vitro,Xinan Nongye Daxue Xuebao(Journal of Southwest Agricultural University),25(6):495-497(周清元,李加纳,殷家明,唐章林,谌利,陶澜,2003,白菜型油菜和羽衣甘蓝种间杂交的初步研究Ⅰ,取材时间对子房离体培养结籽率的影响,西南农业大学学报,25(6):495-497)