黄籽沙逊与黄籽埃塞俄比亚芥种间杂种的鉴定
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摘要
为了把黄籽沙逊的黄籽性状同埃塞俄比亚芥菜的优良特性相结合,在对黄籽沙逊和黄籽埃塞俄比亚芥菜种间远缘杂交的基础上,开展了对种间杂种的形态鉴定、染色体倍性检测、根尖染色体制片和分子标记鉴定研究。形态学鉴定结果表明,黄籽沙逊和埃塞俄比亚芥菜真杂种外观明显不同于其父母本,处于中间类型,更多偏向于父本黄籽埃塞俄比亚芥。总共对11株黄籽沙逊与黄籽埃塞俄比亚芥菜种间杂种后代的倍性进行了检测,流式细胞仪分析结果表明,阳性植株DNA相对含量(主峰位置125)明显小于对照黄籽埃塞俄比亚芥DNA相对含量(主峰位置200),大于亲本黄籽沙逊DNA相对含量(主峰位置75)。细胞学鉴定结果表明,真杂种植株染色体为27条,为黄籽沙逊(AA,2n=20)和黄籽埃塞俄比亚芥菜(BBCC,2n=34)的单倍体染色体相加之和。分子标记鉴定结果表明,真杂种既有父本也有母本的特异性条带。以上均证实了所获得种间杂种的真实性,其包含了双亲中的遗传信息。所获得的含有芸薹属A、B和C 3个亚基因组的种间杂种可作为育种的中间材料应用到甘蓝型油菜的遗传育种研究,具有重要的理论和实践意义。
In order to combine the yellow seed trait of Yellow Sarson with the fine features of Ethiopian mustard,on the basis of interspecific hybridization experiment between Yellow Sarson and yellow seeded Ethiopian mustard,this study carried out morphological identification,chromosome ploidy detection,slide preparation of chromosomes from root tip cells and molecular marker identification for interspecific hybrids. Morphological identification results of hybrid progeny showed that interspecific hybrids between Yellow Sarson and Ethiopia mustard were very different from their parents and belonged to the middle type in appearance,which were more like the maleparent. The results of ploidy analysis by flow cytometry showed that the peak value of positive plants was 125,which was significantly smaller than the main peak of Ethiopia mustard(200),and bigger than the main peak of Yellow Sarson(75). Cytology identification results showed that the real hybrid plants contained 27 chromosomes,which was the sum of the haploid chromosomes number of the Yellow Sarson(AA,2n = 20) and Ethiopia mustard(BBCC,2n = 34). Molec-ular markers identification results showed that true hybrids had specific bands of both male and female parents. The results above indicated that the hybrids were real interspecific hybrids,which contained genetic information from both parents. The created hybrids containing the three subgenomes(A,B and C) of Brassica can be used as intermediate breeding materials for application in genetic breeding research of Brassica napus,which has important theoretical and practical significance.
In order to combine the yellow seed trait of Yellow Sarson with the fine features of Ethiopian mustard,on the basis of interspecific hybridization experiment between Yellow Sarson and yellow seeded Ethiopian mustard,this study carried out morphological identification,chromosome ploidy detection,slide preparation of chromosomes from root tip cells and molecular marker identification for interspecific hybrids. Morphological identification results of hybrid progeny showed that interspecific hybrids between Yellow Sarson and Ethiopia mustard were very different from their parents and belonged to the middle type in appearance,which were more like the maleparent. The results of ploidy analysis by flow cytometry showed that the peak value of positive plants was 125,which was significantly smaller than the main peak of Ethiopia mustard(200),and bigger than the main peak of Yellow Sarson(75). Cytology identification results showed that the real hybrid plants contained 27 chromosomes,which was the sum of the haploid chromosomes number of the Yellow Sarson(AA,2n = 20) and Ethiopia mustard(BBCC,2n = 34). Molec-ular markers identification results showed that true hybrids had specific bands of both male and female parents. The results above indicated that the hybrids were real interspecific hybrids,which contained genetic information from both parents. The created hybrids containing the three subgenomes(A,B and C) of Brassica can be used as intermediate breeding materials for application in genetic breeding research of Brassica napus,which has important theoretical and practical significance.
引文
[1]林超,孙萍,程斐,等.芸薹属植物的远缘杂交[J].山东农业科学,2007(4):27-31.
[2]周庆红,周灿,范淑英.远缘杂交在芸薹属作物育种中的应用研究进展[J].北方园艺,2015(2):165-170.
[3]Prakash S,Chopra V L.Introgression of resistance to shattering in Brassica napus from Brassica juncea through nonhomologous recombination[J].Plant Breeding,1998,101:167-168.
[4]贾思齐,王超.芸薹属作物远缘杂交子房离体培养的研究进展[J].北方农业学报,2016(3):110-114.
[5]杨学乐,陶芬芳,彭烨,等.芸薹属作物远缘杂种优势研究[J].作物研究,2016(5):520-523.
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[7]Kumar A,Singh P,Singh H,et al.Difference in osmo regulation in Brassica species[J].Annals of Botany,1984,54(4):537-541.
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[10]Meng J,Shi S,Gan L,et al.The production of yellow-seeded Brassica napus(AACC)through crossing interspecific hybrids of B.campesrtis(AA)and B.carinata(BBCC)with B.napus[J].Euphytica,1998,103:329-333.
[11]鲁坤存,刘淑艳,郭家保,等.芥甘杂交选育甘蓝型黄籽油菜的研究[J].湖南农业大学学报(自然科学版),2006(2):116-119.
[12]江莹芬,田恩堂,陈伦林,等.埃塞俄比亚芥和白菜型油菜远缘杂种F1的鉴定[J].中国油料作物学报,2007,29(2):103-106.
[13]栗茂腾,张椿雨,李宗芸,等.埃塞俄比亚芥与白菜型油菜间六倍体杂种的获得及其生物学特性研究[J].作物学报,2005,31(12):1579-1585.
[14]仲裕泉,傅寿仲,浦惠明,等.埃塞俄比亚芥黄种皮性状向甘蓝型油菜的转移研究[J].江苏农业学报,1996(2):26-31.
[15]Falk K C.ACS-C29 summer turnip rape[J].Can J Plant Sci,2010,90(2):191-192.
[16]Hutcheson D S,Falk K C,Rakow G F W.TR4 summer turnip rape[J].Can J Plant Sci,2000,80(4):837-838.
[17]Rahman M H,Joersbo M,Poulsen M H.Development of yellow-seed Brassica napus of double low quality[J].Plant Breed,2001,120(6):473-478.
[18]张锦芳,李浩杰,张雪花,等.甘蓝型油菜SSR标记遗传多样性及其与农艺性状关联分析[J].分子植物育种,2013(5):502-512.
[19]杜培,张新友,李丽娜,等.高质量花生根尖细胞染色体制片方法研究[J].河南农业科学,2013,42(3):31-35.
[20]李爱民,张永泰,蒋金金,等.芸苔属主要油料作物黄籽性状分子遗传研究进展[J].分子植物育种,2009(2):407-412.
[21]Daun J K,Declercq R.Quality of yellow and dark seeds in Brassica campestris canola varieties Candle and Tobin[J].J Am Oil Chem Soc,1988,65(1):122-126.
[22]Whetten R W,Mackay J J,Sederoff R.Recent advances in understanding lignin biosynthesis[J].Annu Rev Plant Physiol Plant Mol Biol,1998,49:585-609.
[23]Shirzadegan M,Robbelen G.Influence of seed colour and hull proportions on quality properties of seeds in Brassica napus L.[J].Fette Seifen Anstrichm,1985,87:235-237.
[24]Slominski B A,Campbell L D,Guenter W.Carbohydrates and dietary fibre components of yellow and brown-seeded canola[J].J Agric Food Chem,1994,42:704-707.
[25]Tang Z L,Li J N,Zhang X K,et al.Genetic variation of yellow-seeded rapeseed lines(Brassica napus L.)from different genetic sources[J].Plant Breed,1997,116(5):471-474.
[26]赵德培,张纪增.通过幼胚培养获得结球甘蓝与大白菜的种间杂种[J].中国农业科学,1981,14(2):46-51.
[27]扈新民,李锡香,梁燕,等.甘蓝与萝卜属间杂交及其胚挽救技术优化[J].中国蔬菜,2009(10):7-12.
[28]石淑稳,孟金陵.埃塞俄比亚芥与白菜型、甘蓝型和芥菜型油菜杂种子房培养技术的研究[J].武汉植物学研究,1995(1):8-14.
[29]孟金陵,吴江生,王萍.埃塞俄比亚芥与白菜、芥菜间的可交配性[J].华中农业大学学报,1992,11(3):203-207.
[2]周庆红,周灿,范淑英.远缘杂交在芸薹属作物育种中的应用研究进展[J].北方园艺,2015(2):165-170.
[3]Prakash S,Chopra V L.Introgression of resistance to shattering in Brassica napus from Brassica juncea through nonhomologous recombination[J].Plant Breeding,1998,101:167-168.
[4]贾思齐,王超.芸薹属作物远缘杂交子房离体培养的研究进展[J].北方农业学报,2016(3):110-114.
[5]杨学乐,陶芬芳,彭烨,等.芸薹属作物远缘杂种优势研究[J].作物研究,2016(5):520-523.
[6]熊秋芳,沈金雄,涂金星,等.华中农业大学油菜遗传育种研究五十年[J].中国农业科技导报,2009(6):1-6.
[7]Kumar A,Singh P,Singh H,et al.Difference in osmo regulation in Brassica species[J].Annals of Botany,1984,54(4):537-541.
[8]Malik R S.Prospects for Brassica carinata as an oilseed crop in India[J].Experimental Agriculture,1990,26:125-130.
[9]Rashid A,Rakow G,Downey R K.Development of yellow seeded Brassica napus through interspecific crosses[J].Plant Breeding,1994,112:127-134.
[10]Meng J,Shi S,Gan L,et al.The production of yellow-seeded Brassica napus(AACC)through crossing interspecific hybrids of B.campesrtis(AA)and B.carinata(BBCC)with B.napus[J].Euphytica,1998,103:329-333.
[11]鲁坤存,刘淑艳,郭家保,等.芥甘杂交选育甘蓝型黄籽油菜的研究[J].湖南农业大学学报(自然科学版),2006(2):116-119.
[12]江莹芬,田恩堂,陈伦林,等.埃塞俄比亚芥和白菜型油菜远缘杂种F1的鉴定[J].中国油料作物学报,2007,29(2):103-106.
[13]栗茂腾,张椿雨,李宗芸,等.埃塞俄比亚芥与白菜型油菜间六倍体杂种的获得及其生物学特性研究[J].作物学报,2005,31(12):1579-1585.
[14]仲裕泉,傅寿仲,浦惠明,等.埃塞俄比亚芥黄种皮性状向甘蓝型油菜的转移研究[J].江苏农业学报,1996(2):26-31.
[15]Falk K C.ACS-C29 summer turnip rape[J].Can J Plant Sci,2010,90(2):191-192.
[16]Hutcheson D S,Falk K C,Rakow G F W.TR4 summer turnip rape[J].Can J Plant Sci,2000,80(4):837-838.
[17]Rahman M H,Joersbo M,Poulsen M H.Development of yellow-seed Brassica napus of double low quality[J].Plant Breed,2001,120(6):473-478.
[18]张锦芳,李浩杰,张雪花,等.甘蓝型油菜SSR标记遗传多样性及其与农艺性状关联分析[J].分子植物育种,2013(5):502-512.
[19]杜培,张新友,李丽娜,等.高质量花生根尖细胞染色体制片方法研究[J].河南农业科学,2013,42(3):31-35.
[20]李爱民,张永泰,蒋金金,等.芸苔属主要油料作物黄籽性状分子遗传研究进展[J].分子植物育种,2009(2):407-412.
[21]Daun J K,Declercq R.Quality of yellow and dark seeds in Brassica campestris canola varieties Candle and Tobin[J].J Am Oil Chem Soc,1988,65(1):122-126.
[22]Whetten R W,Mackay J J,Sederoff R.Recent advances in understanding lignin biosynthesis[J].Annu Rev Plant Physiol Plant Mol Biol,1998,49:585-609.
[23]Shirzadegan M,Robbelen G.Influence of seed colour and hull proportions on quality properties of seeds in Brassica napus L.[J].Fette Seifen Anstrichm,1985,87:235-237.
[24]Slominski B A,Campbell L D,Guenter W.Carbohydrates and dietary fibre components of yellow and brown-seeded canola[J].J Agric Food Chem,1994,42:704-707.
[25]Tang Z L,Li J N,Zhang X K,et al.Genetic variation of yellow-seeded rapeseed lines(Brassica napus L.)from different genetic sources[J].Plant Breed,1997,116(5):471-474.
[26]赵德培,张纪增.通过幼胚培养获得结球甘蓝与大白菜的种间杂种[J].中国农业科学,1981,14(2):46-51.
[27]扈新民,李锡香,梁燕,等.甘蓝与萝卜属间杂交及其胚挽救技术优化[J].中国蔬菜,2009(10):7-12.
[28]石淑稳,孟金陵.埃塞俄比亚芥与白菜型、甘蓝型和芥菜型油菜杂种子房培养技术的研究[J].武汉植物学研究,1995(1):8-14.
[29]孟金陵,吴江生,王萍.埃塞俄比亚芥与白菜、芥菜间的可交配性[J].华中农业大学学报,1992,11(3):203-207.