芥蓝和结球甘蓝分别与甘蓝型油菜杂交的后代结实性及Rfo传递效率比较
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摘要
为明确甘蓝与甘蓝型油菜种间杂交的生殖隔离程度是否与甘蓝的变种类型有关,以2份Ogura CMS芥蓝和4份Ogura CMS结球甘蓝为母本,分别与2份含有Ogura CMS育性恢复基因Rfo的甘蓝型油菜杂交,获得的种间杂种加倍后,再分别与母本芥蓝和结球甘蓝回交,调查F_1、BC_1两个世代材料的结实性和Rfo基因的传递效率。结果表明,芥蓝与甘蓝型油菜种间杂交F_1代、BC_1代的亲和指数都显著高于结球甘蓝(P<0.05),是结球甘蓝的2倍以上,且在芥蓝材料中Rfo基因的传递效率高于结球甘蓝。进一步比较BC_1回交后代Rfo阳性单株的育性、遗传背景回复率,芥蓝的育性大多显著好于结球甘蓝(P<0.05),26株BC_1单株遗传背景回复率在0.50~0.84之间;而结球甘蓝两个BC_1单株(15Y1和15Y4)遗传背景回复率分别为0.53和0.59。上述结果说明,芥蓝较结球甘蓝更易与甘蓝型油菜杂交获得后代,且异源目的片段的传递效率更高,因此芥蓝可作为结球甘蓝与甘蓝型油菜杂交的桥梁材料,加速甘蓝型油菜中优异性状的转育进程。
In order to determine whether the seed-setting of distant hybridization between Brassica napus and B. oleracea is affected by different B. oleracea sub-species or not,in this study,two Chinese kale and four cabbage materials with Ogura CMS cytoplasm were crossed with two homozygous rapeseed restored lines harboring the Ogura CMS fertility-restored gene Rfo. The F_1 interspecific hybrid plants with doubled chromosome were backcrossed with Ogura CMS Chinese kale and cabbage to produce BC_1 plants,respectively. The seed-setting and the transmission rate of Rfo of Chinese kale × rapeseed cross combinations(CKR)and cabbage × rapeseed cross combinations(CR)were investigated in two generations,respectively. The results suggested that the compatibility index of CKR was two folds higher than that of CR,with significant difference between CKR and CR in two generations(P < 0.05). In addition,the Rfo gene transmission rate of CKR was higher than that of CR. The pollen viability and recovering ratio of genetic background(RRGB)of all BC_1 Rfo-positive individuals obtained from CKR and CR were assessed. The pollen viability of most BC_1 Rfo-positive individuals obtained from CKR were significantly higher than that of the two Rfo-positive individuals obtained from CR(P < 0.05). The RRGB of 26 Rfo-positive individuals obtained from CKR were 0.50–0.84. While the RRGB of two Rfo-positive individuals(15 Y1 and 15 Y4)obtained from CR were 0.53 and 0.59,respectively. Our results revealed that compared with cabbage,Chinese kale is much easier to produce progenies when crossed with B. napus and the transmission rate of alien fragment was much higher. Therefore,Chinese kale could be used as a bridge to accelerate the process of the beneficial agronomic traits transfer from B. napus to cabbage.
In order to determine whether the seed-setting of distant hybridization between Brassica napus and B. oleracea is affected by different B. oleracea sub-species or not,in this study,two Chinese kale and four cabbage materials with Ogura CMS cytoplasm were crossed with two homozygous rapeseed restored lines harboring the Ogura CMS fertility-restored gene Rfo. The F_1 interspecific hybrid plants with doubled chromosome were backcrossed with Ogura CMS Chinese kale and cabbage to produce BC_1 plants,respectively. The seed-setting and the transmission rate of Rfo of Chinese kale × rapeseed cross combinations(CKR)and cabbage × rapeseed cross combinations(CR)were investigated in two generations,respectively. The results suggested that the compatibility index of CKR was two folds higher than that of CR,with significant difference between CKR and CR in two generations(P < 0.05). In addition,the Rfo gene transmission rate of CKR was higher than that of CR. The pollen viability and recovering ratio of genetic background(RRGB)of all BC_1 Rfo-positive individuals obtained from CKR and CR were assessed. The pollen viability of most BC_1 Rfo-positive individuals obtained from CKR were significantly higher than that of the two Rfo-positive individuals obtained from CR(P < 0.05). The RRGB of 26 Rfo-positive individuals obtained from CKR were 0.50–0.84. While the RRGB of two Rfo-positive individuals(15 Y1 and 15 Y4)obtained from CR were 0.53 and 0.59,respectively. Our results revealed that compared with cabbage,Chinese kale is much easier to produce progenies when crossed with B. napus and the transmission rate of alien fragment was much higher. Therefore,Chinese kale could be used as a bridge to accelerate the process of the beneficial agronomic traits transfer from B. napus to cabbage.
引文
Ayotte R,Harney P M,Machado V S.1987.The transfer of triazine resistance from Brassica napus L.to B.oleracea L.Ⅰ.Production of F1 hybrids through embryo rescue.Euphytica,36:615-624.
Bannerot H,Boulidard L,Couderon Y,Temple J.1974.Transfer of cytoplasmic male sterility from Raphanus sativus to Brassica oleracea//Wills AB,North C.Proc eucarpia meet cruciferae.Scottish Hortic Res Inst,Invergavrie,52-54.
Budahn H,Schrader O,Peterka H.2008.Development of a complete set of disomic rape-radish chromosome-addition line.Euphytica,162:117-128.
Dai Hua-jun,Zhu Zheng-bin,Shen Xue-lin,Zhou Jian-ming,He Jian-hua.2010.Essences and approaches of distant hybridization in crops breeding.Genomics and Applied Biology,29(1):144-149.(in Chinese)戴华军,朱正斌,沈雪林,周建明,何建华.2010.作物远缘杂交育种的途径及其实质.基因组学与应用生物学,29(1):144-149.
Deng Jing,Li Xun.2006.The research of incompatibility of distant hybridzation in Brassica crops and genetic characteristics of its hybrids.Crop Research,20(5):459-464.(in Chinese)邓婧,李栒.2006.芸薹属植物远缘杂交不亲和性以及杂种后代遗传特性的研究进展.作物研究,20(5):459-464.
Ding L,Zhao Z G,Ge X H,Li Z Y.2013.Intergeneric addition and substitution of Brassica napus with different chromosomes from Orychophragmus violaceus:Phenotype and cytology.Scientia Horticulturae,164:303-309.
Dolezel J,Greilhuber J,Suda J.2007.Estimation of nuclear DNA content in plants using flow cytometry.Nat Protoc,2:2233-2244.
Dong Dong.2011.Studies on SSR marker assisted selection for genetic background in wheat[Ph.D.Dissertation].Beijing:Chinese Academy of Agricultural Sciences.(in Chinese)董冬.2011.小麦SSR标记辅助遗传背景选择技术研究[博士论文].北京:中国农业科学院.
Fehr W R.1987.Principles of cultivar development,Vol.1 Theory and technique.New York:Macmillan Pub.Co.
Frisch M,Bohn M,Melchinger A E.1999.Comparison of selection strategies for marker-assisted backcrossing of a gene.Crop Sci,39:1295-1301.
Hospital F C,Chevalet P M.1992.Using markers in gene introgression breeding programes.Genetics,132:1199-1210.
Kalloo G.1992.Utilization of wild species//Kalloo G,Chawdhury J B.Distant hybridiza tion of crop plants.Springer,Berlin,149-167.
Li Qin-fei,Qian Wei,He Ya-jun.2016.Crossability between Brassica napus and B.oleracea Journal of Southwestern University(Natural Science Edition),38(10):1-7.(in Chinese)李勤菲,钱伟,贺亚军.2016.甘蓝型油菜与甘蓝杂交的亲和性分析.西南大学学报(自然科学版),38(10):1-7.
Li Q F,Mei J Q,Zhang Y J,Li J N,Ge X H,Li Z Y,Qian W.2013.A large-scale introgression of genomic components of Brassica rapa into B.napus by the bridge of hexaploid derived from between B.napus and B.oleracea.Theor Appl Genet,126(8):2073-2080.
Liu Yao,Ding Yi-juan,Wang Lei,Wan Hua-fang,Mei Jia-qin,Qian Wei.2015.Crossability between Brassica napus with hexaploid AnAnCnCnCoCo and sclerotinia resistance in the hybrids.Scientia Agricultura Sinica,48(24):4885-4891.(in Chinese)刘瑶,丁一娟,汪雷,万华芳,梅家琴,钱伟.2015.甘蓝型油菜与AnAnCnCnCoCo六倍体的可交配性及杂交种菌核病抗性.中国农业科学,48(24):4885-4891.
Lin Chao,Sun Ping,Cheng Fei,Wang Lei,Gao Jian-wei.2007.Distant hybridzation in Brassica crops.Shandong Agricultural Sciences,4:27-31.(in Chinese)林超,孙萍,程斐,王磊,高建伟.2007.芸薹属植物的远缘杂交.山东农业科学,4:27-31.
Mei J,Liu Y,Wei D,Wittkop B,Ding Y,Li Q,Li J,Wan H,Li Z,Ge X,Frauen M,Snowdon R,Qian W,Friedtet W.2015.Transfer of sclerotinia resistance from wild relative of Brassica oleracea into Brassica napus using a hexaploidy step.Theor Appl Genet,128:639-644.
Mei Jia-qin.2011.Genetic investigation of relationships between Brassica oleracea and C subgenome of Brassca napus and mapping QTL for sclerotinia resistance in B.oleracea[Ph.D.Dissertation].Chongqing:Southwest University.(in Chinese)梅家琴.2011.甘蓝与甘蓝型油菜C亚基因组遗传关系调查及甘蓝抗菌核病QTL定位[博士论文].重庆:西南大学.
Meng Jin-ling.1987.The research progress of intergeneric incompatibility in Brassica crops.Oil Crops,4:71-77.(in Chinese)孟金陵.1987.芸薹属植物属间杂交不亲和性研究进展.中国油料,4:71-77.
Nasrallah J B,Liu P,Sherman-Broyles S,Schmidt R,Nasrallah M E.2007.Epigenetic mechanisms for breakdown of self-incompatibility in interspecific hybrids.Genetics,175:1965-1973.
Pratap A,Kumar J.2014.Alien gene transfer in crop plants.Volume 2.Springer:New York:121-151.
Primard-Brisset C,Poupard JP,Horvais R,Eber F,Pelletier G,Remard M,Delourme R.2005.A new recombined double low restorer line for the Ogu-INRA cms in rapeseed(Brassica napus L.).Theor Appl Genet,111:736-746.
Ripley V L,Beversdorf W D.2003.Development of self-incompatible Brassica napus(Ⅰ)introgression of S-alleles from Brassica oleracea through interspecific hybridization.Plant Breed,122:1-5.
Saal B,Brun H,Glais I,Struss D.2004.Identification of a Brassica juncea-derived recessive gene conferring resistance to Leptosphaeria maculans in oilseed rape.Plant Breed,123:505-511.
Saghai-Maroof M A,Soliman K M,Jorgensen R A,Allard R W.1984.Ribosomal DNA spacer-length polymorphisms in barley:Mendelian inheritance,chromosomal location,and population dynamics.Proc Natl Acad Sci USA,81:8014-8018.
Tanksley S D,Young N D,Patterson A H,Bonierbale M W.1989.RFLP mapping in plant breeding:New tools for an old science.Bio/Technology,7:257-263.
Wen J,Tu J X,Li Z Y,Fu T D,Ma C Z,Shen J X.2008.Improving ovary and embryo culture techniques for efficient resynthesis of Brassica napus from reciprocal crosses between yellow-seeded diploids B.rapa and B.oleracea.Euphytica,162:81-89.
Xu Shu-fang,Xuan Zheng-ying,Feng Hui.2004.Compatibility of interspecific hybridzation in Brassica crops.China Veg,1(3):28-29.(in Chinese)徐书法,轩正英,冯辉.2004.芸薹属农作物种间杂交的亲和性.中国蔬菜,1(3):28-29.
Yang Yu-zhu,Zhang Xiao-lan,Yang Wei-guo,Tian Wen-wei,Wang Qin.2011.Studies on the resistance of rape variety to Plasmodiophora brassicae and screening.Chinese Agricultural Science Bulletin,27(16):278-282.(in Chinese)杨玉珠,张晓兰,杨卫国,田文伟,王勤.2011.油菜抗根肿病品种筛选试验研究.中国农学通报,27(16):278-282.
Yu Hai-long,Fang Zhi-yuan,Liu Yu-mei,,Yang Li-mei,Zhuang Mu,LüHong-hao,Li Zhan-sheng,Zhang Yang-yong.2015.Genetic background screen of inter-specific hybrids between SSR marker assisted Chinese kale and rapeseed.China Veg,8:14-21.(in Chinese)于海龙,方智远,刘玉梅,杨丽梅,庄木,吕红豪,李占省,张扬勇.2015.SSR标记辅助芥蓝×甘蓝型油菜种间杂交后代的遗传背景筛选.中国蔬菜,8:14-21.
Yu H L,Fang Z Y,Liu Y M,Yang L M,Zhuang M,LüH H,Li Z S,Han F Q,Liu X P,Zhang Y Y.2016.Development of a novel allele-specific Rfo marker and creation of Ogura CMS fertility-restored interspecific hybrids in Brassica oleracea.Theor Appl Genet,129:1625-1637.
Zhang G Q,Tang G X,Song W J,Zhou W J.2004.Resynthesizing Brassica napus from interspecific hybridization between Brassica rapa and B.oleracea through ovary culture.Euphytica,140:181-187.
Zhang Yang-yong.2011.Research on the cytoplasmic DNA diversity in Brassica oleracea vegetables[Ph.D.Dissertation].Beijing:Chinese Academy of Agricultural Sciences.(in Chinese)张扬勇.2011.甘蓝类蔬菜的细胞质DNA多样性研究[博士论文].北京:中国农业科学院.
Zhu Peng-fang,Wang Xing,Zhang Jian,Wei Yu-tang.2011.Studies on the compatibility of backcrosses of interspecific and inter varietal hybrids in Brassica.Chinese Agricultural Science Bulletin,27(10):149-152.(in Chinese)祝朋芳,王兴,张健,魏毓棠.2011.芸薹属种与变种间杂交后代回交亲和性的研究.中国农学通报,27(10):149-152.
Bannerot H,Boulidard L,Couderon Y,Temple J.1974.Transfer of cytoplasmic male sterility from Raphanus sativus to Brassica oleracea//Wills AB,North C.Proc eucarpia meet cruciferae.Scottish Hortic Res Inst,Invergavrie,52-54.
Budahn H,Schrader O,Peterka H.2008.Development of a complete set of disomic rape-radish chromosome-addition line.Euphytica,162:117-128.
Dai Hua-jun,Zhu Zheng-bin,Shen Xue-lin,Zhou Jian-ming,He Jian-hua.2010.Essences and approaches of distant hybridization in crops breeding.Genomics and Applied Biology,29(1):144-149.(in Chinese)戴华军,朱正斌,沈雪林,周建明,何建华.2010.作物远缘杂交育种的途径及其实质.基因组学与应用生物学,29(1):144-149.
Deng Jing,Li Xun.2006.The research of incompatibility of distant hybridzation in Brassica crops and genetic characteristics of its hybrids.Crop Research,20(5):459-464.(in Chinese)邓婧,李栒.2006.芸薹属植物远缘杂交不亲和性以及杂种后代遗传特性的研究进展.作物研究,20(5):459-464.
Ding L,Zhao Z G,Ge X H,Li Z Y.2013.Intergeneric addition and substitution of Brassica napus with different chromosomes from Orychophragmus violaceus:Phenotype and cytology.Scientia Horticulturae,164:303-309.
Dolezel J,Greilhuber J,Suda J.2007.Estimation of nuclear DNA content in plants using flow cytometry.Nat Protoc,2:2233-2244.
Dong Dong.2011.Studies on SSR marker assisted selection for genetic background in wheat[Ph.D.Dissertation].Beijing:Chinese Academy of Agricultural Sciences.(in Chinese)董冬.2011.小麦SSR标记辅助遗传背景选择技术研究[博士论文].北京:中国农业科学院.
Fehr W R.1987.Principles of cultivar development,Vol.1 Theory and technique.New York:Macmillan Pub.Co.
Frisch M,Bohn M,Melchinger A E.1999.Comparison of selection strategies for marker-assisted backcrossing of a gene.Crop Sci,39:1295-1301.
Hospital F C,Chevalet P M.1992.Using markers in gene introgression breeding programes.Genetics,132:1199-1210.
Kalloo G.1992.Utilization of wild species//Kalloo G,Chawdhury J B.Distant hybridiza tion of crop plants.Springer,Berlin,149-167.
Li Qin-fei,Qian Wei,He Ya-jun.2016.Crossability between Brassica napus and B.oleracea Journal of Southwestern University(Natural Science Edition),38(10):1-7.(in Chinese)李勤菲,钱伟,贺亚军.2016.甘蓝型油菜与甘蓝杂交的亲和性分析.西南大学学报(自然科学版),38(10):1-7.
Li Q F,Mei J Q,Zhang Y J,Li J N,Ge X H,Li Z Y,Qian W.2013.A large-scale introgression of genomic components of Brassica rapa into B.napus by the bridge of hexaploid derived from between B.napus and B.oleracea.Theor Appl Genet,126(8):2073-2080.
Liu Yao,Ding Yi-juan,Wang Lei,Wan Hua-fang,Mei Jia-qin,Qian Wei.2015.Crossability between Brassica napus with hexaploid AnAnCnCnCoCo and sclerotinia resistance in the hybrids.Scientia Agricultura Sinica,48(24):4885-4891.(in Chinese)刘瑶,丁一娟,汪雷,万华芳,梅家琴,钱伟.2015.甘蓝型油菜与AnAnCnCnCoCo六倍体的可交配性及杂交种菌核病抗性.中国农业科学,48(24):4885-4891.
Lin Chao,Sun Ping,Cheng Fei,Wang Lei,Gao Jian-wei.2007.Distant hybridzation in Brassica crops.Shandong Agricultural Sciences,4:27-31.(in Chinese)林超,孙萍,程斐,王磊,高建伟.2007.芸薹属植物的远缘杂交.山东农业科学,4:27-31.
Mei J,Liu Y,Wei D,Wittkop B,Ding Y,Li Q,Li J,Wan H,Li Z,Ge X,Frauen M,Snowdon R,Qian W,Friedtet W.2015.Transfer of sclerotinia resistance from wild relative of Brassica oleracea into Brassica napus using a hexaploidy step.Theor Appl Genet,128:639-644.
Mei Jia-qin.2011.Genetic investigation of relationships between Brassica oleracea and C subgenome of Brassca napus and mapping QTL for sclerotinia resistance in B.oleracea[Ph.D.Dissertation].Chongqing:Southwest University.(in Chinese)梅家琴.2011.甘蓝与甘蓝型油菜C亚基因组遗传关系调查及甘蓝抗菌核病QTL定位[博士论文].重庆:西南大学.
Meng Jin-ling.1987.The research progress of intergeneric incompatibility in Brassica crops.Oil Crops,4:71-77.(in Chinese)孟金陵.1987.芸薹属植物属间杂交不亲和性研究进展.中国油料,4:71-77.
Nasrallah J B,Liu P,Sherman-Broyles S,Schmidt R,Nasrallah M E.2007.Epigenetic mechanisms for breakdown of self-incompatibility in interspecific hybrids.Genetics,175:1965-1973.
Pratap A,Kumar J.2014.Alien gene transfer in crop plants.Volume 2.Springer:New York:121-151.
Primard-Brisset C,Poupard JP,Horvais R,Eber F,Pelletier G,Remard M,Delourme R.2005.A new recombined double low restorer line for the Ogu-INRA cms in rapeseed(Brassica napus L.).Theor Appl Genet,111:736-746.
Ripley V L,Beversdorf W D.2003.Development of self-incompatible Brassica napus(Ⅰ)introgression of S-alleles from Brassica oleracea through interspecific hybridization.Plant Breed,122:1-5.
Saal B,Brun H,Glais I,Struss D.2004.Identification of a Brassica juncea-derived recessive gene conferring resistance to Leptosphaeria maculans in oilseed rape.Plant Breed,123:505-511.
Saghai-Maroof M A,Soliman K M,Jorgensen R A,Allard R W.1984.Ribosomal DNA spacer-length polymorphisms in barley:Mendelian inheritance,chromosomal location,and population dynamics.Proc Natl Acad Sci USA,81:8014-8018.
Tanksley S D,Young N D,Patterson A H,Bonierbale M W.1989.RFLP mapping in plant breeding:New tools for an old science.Bio/Technology,7:257-263.
Wen J,Tu J X,Li Z Y,Fu T D,Ma C Z,Shen J X.2008.Improving ovary and embryo culture techniques for efficient resynthesis of Brassica napus from reciprocal crosses between yellow-seeded diploids B.rapa and B.oleracea.Euphytica,162:81-89.
Xu Shu-fang,Xuan Zheng-ying,Feng Hui.2004.Compatibility of interspecific hybridzation in Brassica crops.China Veg,1(3):28-29.(in Chinese)徐书法,轩正英,冯辉.2004.芸薹属农作物种间杂交的亲和性.中国蔬菜,1(3):28-29.
Yang Yu-zhu,Zhang Xiao-lan,Yang Wei-guo,Tian Wen-wei,Wang Qin.2011.Studies on the resistance of rape variety to Plasmodiophora brassicae and screening.Chinese Agricultural Science Bulletin,27(16):278-282.(in Chinese)杨玉珠,张晓兰,杨卫国,田文伟,王勤.2011.油菜抗根肿病品种筛选试验研究.中国农学通报,27(16):278-282.
Yu Hai-long,Fang Zhi-yuan,Liu Yu-mei,,Yang Li-mei,Zhuang Mu,LüHong-hao,Li Zhan-sheng,Zhang Yang-yong.2015.Genetic background screen of inter-specific hybrids between SSR marker assisted Chinese kale and rapeseed.China Veg,8:14-21.(in Chinese)于海龙,方智远,刘玉梅,杨丽梅,庄木,吕红豪,李占省,张扬勇.2015.SSR标记辅助芥蓝×甘蓝型油菜种间杂交后代的遗传背景筛选.中国蔬菜,8:14-21.
Yu H L,Fang Z Y,Liu Y M,Yang L M,Zhuang M,LüH H,Li Z S,Han F Q,Liu X P,Zhang Y Y.2016.Development of a novel allele-specific Rfo marker and creation of Ogura CMS fertility-restored interspecific hybrids in Brassica oleracea.Theor Appl Genet,129:1625-1637.
Zhang G Q,Tang G X,Song W J,Zhou W J.2004.Resynthesizing Brassica napus from interspecific hybridization between Brassica rapa and B.oleracea through ovary culture.Euphytica,140:181-187.
Zhang Yang-yong.2011.Research on the cytoplasmic DNA diversity in Brassica oleracea vegetables[Ph.D.Dissertation].Beijing:Chinese Academy of Agricultural Sciences.(in Chinese)张扬勇.2011.甘蓝类蔬菜的细胞质DNA多样性研究[博士论文].北京:中国农业科学院.
Zhu Peng-fang,Wang Xing,Zhang Jian,Wei Yu-tang.2011.Studies on the compatibility of backcrosses of interspecific and inter varietal hybrids in Brassica.Chinese Agricultural Science Bulletin,27(10):149-152.(in Chinese)祝朋芳,王兴,张健,魏毓棠.2011.芸薹属种与变种间杂交后代回交亲和性的研究.中国农学通报,27(10):149-152.