MAS育种改良籼稻恢复系R747及其杂交种稻瘟病抗性
|
摘要
为了改良籼稻恢复系R747及其杂交种的稻瘟病抗性,以携带广谱抗稻瘟病基因Pi9的籼稻品系75-1-127为供体亲本,以R747为受体及轮回亲本,利用Pi9基因内功能标记,开展分子标记辅助选择育种实践。获得了如下主要结果:室内接种试验表明,75-1-127与R747对来自国内外不同稻区的24份稻瘟菌菌株的抗菌谱差异显著,抗性频率分别为91.7%和41.7%,两亲本在田间自然病圃苗瘟抗性表现分别为高抗和高感;根据Pi9基因序列信息开发了一个共显性多态标记SPL-1,该标记在75-1-127和R747基因组中分别扩增出一条745 bp和一条850 bp的条带,标记基因型对稻瘟病抗感表型的选择效率为100%;从BC6F3株系中遴选出一个高抗苗瘟的改良恢复系R747-Pi9,并用它与不育系培矮64S、P88S和桃1A分别配制出高抗稻瘟病的杂交种;利用分布于水稻全基因组不同位点的220个SSR标记,分析了改良抗病恢复系R747-Pi9与受体亲本R747的遗传背景差异,结果表明R747-Pi9对R747的背景回复率为0.94。本研究通过分子标记辅助选择和连续回交育种,成功定向改良了恢复系R747及其杂交种的稻瘟病抗性。
To improve blast resistance of the indica rest orer line R747 and its hybrids, the indica rice line75-1-127 carrying a broad spectrum rice blast resistance gene Pi9 was used as the donor parent and R747 as the recipient and recurrent parent. Functional markers of Pi9 gene were used to carry out molecular marker-assisted selection breeding. The main results obtained were as follows: Greenhouse inoculation showed that resistance spectra to 24 tested worldwide M. oryzae isolates were obviously different between 75-1-127 and R747, with resistance frequencies of 91.7% and 41.7% respectively. Moreover, two parents showed high resistance and high susceptibility to blast in natural field nursery, respectively. Based on the sequence information of Pi9 gene, a co-dominant polymorphic marker SPL-1 was developed. A 745 bp band was amplified from 75-1-127 genome while a 850 bp band from R747, and the selection efficiency by SPL-1 genotype for blast resistance phenotype reached 100%. A seedling blast resistance improved restorer 'R747-Pi9' was bred from the BC6 F3 strains, and hybrids developed by R747-Pi9 and three rice sterile lines Peiai64 S, P88 S and Tao1 A also showed high-level blast resistance in field nursery. Using 220 SSR markers distributed in different rice genome locations, the genetic background differences between the improved blast resistant restorer line R747-Pi9 and the recipient parent R747 were analyzed, and the results indicated that the background response rate of R747-Pi9 to R747 was 0.94. In this study, the rice blast resistance of the indica restorer R747 and its hybrids was successfully improved through molecular marker-assisted selection and continuous backcross breeding.
To improve blast resistance of the indica rest orer line R747 and its hybrids, the indica rice line75-1-127 carrying a broad spectrum rice blast resistance gene Pi9 was used as the donor parent and R747 as the recipient and recurrent parent. Functional markers of Pi9 gene were used to carry out molecular marker-assisted selection breeding. The main results obtained were as follows: Greenhouse inoculation showed that resistance spectra to 24 tested worldwide M. oryzae isolates were obviously different between 75-1-127 and R747, with resistance frequencies of 91.7% and 41.7% respectively. Moreover, two parents showed high resistance and high susceptibility to blast in natural field nursery, respectively. Based on the sequence information of Pi9 gene, a co-dominant polymorphic marker SPL-1 was developed. A 745 bp band was amplified from 75-1-127 genome while a 850 bp band from R747, and the selection efficiency by SPL-1 genotype for blast resistance phenotype reached 100%. A seedling blast resistance improved restorer 'R747-Pi9' was bred from the BC6 F3 strains, and hybrids developed by R747-Pi9 and three rice sterile lines Peiai64 S, P88 S and Tao1 A also showed high-level blast resistance in field nursery. Using 220 SSR markers distributed in different rice genome locations, the genetic background differences between the improved blast resistant restorer line R747-Pi9 and the recipient parent R747 were analyzed, and the results indicated that the background response rate of R747-Pi9 to R747 was 0.94. In this study, the rice blast resistance of the indica restorer R747 and its hybrids was successfully improved through molecular marker-assisted selection and continuous backcross breeding.
引文
Amante-Bordeos A.,Sitch L.A.,Nelson R.,Dalmacio R.D.,Oliva N.P.,Aswidinnoor H.,and Leung H.,1992,Transfer of bacterial blight and blast resistance from the tetraploid wild rice Oryza minuta to cultivated rice,Oryza sativa,Theor.Appl.Genet.,84(3-4):345-354
Bai B.,Wu J.,Zhou B.,and Deng Q.Y.,2012,Research progress of molecular breeding of rice blast resistance,Zajiao Shuidao(Hybrid Rice),27(3):5-9(柏斌,吴俊,周波,邓启云,2012,稻瘟病抗性分子育种研究综述,杂交水稻,27(3):5-9)
Bonman J.M.,VergeldeDios T.L.,and Khin M.M.,1986,Physiologic specialization of Pyricularia oryzae in the Philippines,Plant Disease,70(8):767-769
Cao Z.,Zeng G.,Hao M.,Sheng H.W.,Ye N.Z.,and Xiao Y.H.,2015,Improving blast resistance of dual-purpose genic sterile line C815S by using molecular marker-assisted selection,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(6):1193-1200(曹志,曾盖,郝明,盛浩闻,叶乃忠,肖应辉,2015,利用MAS技术改良水稻两用核不育系C815S的稻瘟病抗性,分子植物育种,13(6):1193-1200)
Chen H.L.,Li Y.C.,Liu X.L.,Liu J.L.,Yang F.Y.,Xing X.,Liao H.,Huang J.,and Liu J.F.,2017,Improving Blast Resistance of indica rice restorer R599 and its two-line hybrid using MAS technology,Zajiao Shuidao(Hybrid Rice),32(5):61-65(陈海龙,李永聪,刘雄伦,刘金灵,杨丰宇,行璇,廖花,黄俊,刘建丰,2017,MAS育种改良籼稻恢复系R599及其杂交种的稻瘟病抗性,杂交水稻,32(5):61-65)
Li Y.,Nie Y.Y.,Cai Y.H.,Yan M.L.,Mao L.H.,Li Y.H.,Lu M.,and Li M.D.,2011,Research progress in rice blast resistant genes in rice and their molecular marker assisted selection breeding,Jiangxi Nongye Xuebao(Acta Agriculturae Jiangxi),23(8):115-117(李瑶,聂元元,蔡耀辉,颜满莲,毛凌华,李永辉,卢明,李名迪,2011,水稻稻瘟病抗性基因及其分子标记辅助选择育种研究进展,江西农业学报,23(8):115-117)
Liu G.,Lu G.,Zeng L.,and Wang G.L.,2002,Two broad-spectrum blast resistance genes,Pi9(t)and Pi2(t),are physically linked on rice chromosome 6,Mol.Genet.Genomics,267(4):472-480
LüX.L.,Bai H.B.,Hui J.,Cai Z.Y.,Zhang L.F.,and Li S.H.,2017,The improvement of rice blast resistance by broadspectrum gene Pigm,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(11):4336-4342(吕学莲,白海波,惠建,蔡正云,张龙飞,李树华,2017,利用广谱抗病基因Pigm改良水稻稻瘟病抗性的研究,分子植物育种,15(11):4336-4342)
Moffat A.S.,1994,Plant genetics,mapping the sequence of disease resistance,Science,265(5180):1804-1805
Murray M.G.,and Thompson W.F.,1980,Rapid isolation of high molecular weight plant DNA,Nucleic Acids Res.,8(19):4321-4326
Ni D.H.,Yi C.X.,Li L.,Wang X.F.,Wang W.X.,and Yang J.B.,2005,Pyramiding Xa21 and Pi9(t)in rice by marker-assisted selection,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),3(3):329-334(倪大虎,易成新,李莉,汪秀峰,王文相,杨剑波,2005,利用分子标记辅助选择聚合水稻基因Xa21和Pi9(t),分子植物育种,3(3):329-334)
Qu S.H.,Liu G.F.,Zhou B.,Bellizzi M.,Zeng L.R.,Dai L.Y.,Han B.,and Wang G.L.,2006,The broad-spectrum blast resistance gene Pi9 encodes a nucleotide-binding site-leucinerich repeat protein and is a member of a multigene family in rice,Genetics,172(3):1901-1914
Tan L.Q.,Liu X.L.,Yang T.T.,Xing X.,Liang Y.,Chen H.L.,Yang F.Y.,Li Y.C.,Liu J.L.,Dai L.Y.,Chen L.Y.,and Wang G.L.,2015,Breeding new rice line with blast resistance by molecular marker-assisted selection of the Pi9gene,Zuowu Yanjiu(Crop Research),29(4):348-351,356(谭令辞,刘雄伦,杨婷婷,行璇,梁毅,陈海龙,杨丰宇,李永聪,刘金灵,戴良英,陈立云,王国梁,2015,利用Pi9基因分子标记辅助选择培育抗稻瘟病水稻新品系,作物研究,29(4):348-351,356)
Xiao G.,Borja F.N.,Mauleon R.,Padilla J.,Telebanco-Yanoria M.J.,Yang J.,Lu G.D.,Dionisio-Sese M.,and Zhou Bo.,2017,Identification of resistant germplasm containing novel resistance genes at or tightly linked to the Pi2/9 locus conferring broad-spectrum resistance against rice blast,Rice,10(1):37
Xing X.,Liu X.L.,Chen H.L.,Yang F.Y.,Li Y.C.,Liao H.,You L.,Liu J.L.,Dai L.Y.,and Wang G.L.,2016,Improving blast resistance of rice restorer R288 by molecular marker-assisted selection of Pi9 gene,Zuowu Yanjiu(Crop Research),30(5):487-491(行璇,刘雄伦,陈海龙,杨丰宇,李永聪,廖花,游亮,刘金灵,戴良英,王国梁,2016,分子标记辅助选择Pi9基因改良R288的稻瘟病抗性,作物研究,30(5):487-491)
Yang F.Y.,Li Y.C.,Liu X.L.,Liu J.L.,Chen H.L.,Liao H.,Huang J.,and Liang Y.,2017,Improving blast resistance of the early indica rice 1701 by molecular marker-assisted selection,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(6):2212-2217(杨丰宇,李永聪,刘雄伦,刘金灵,陈海龙,廖花,黄俊,梁毅,2017,分子标记辅助选择改良早籼稻1701的稻瘟病抗性,分子植物育种,15(6):2212-2217)
Yang T.T.,Liu X.L.,Tan L.Q.,Xing X.,Wen T.,Liang Y.,Wu J.,Liu J.L.,Dai L.Y.,Liu J.F.,and Wang G.L.,2014,Development of the Pi9 gene marker and its breeding practice in rice blast resistance,Zuowu Yanjiu(Crop Research),28(3):231-235(杨婷婷,刘雄伦,谭令辞,行璇,文婷,梁毅,吴俊,刘金灵,戴良英,刘建丰,王国梁,2014,水稻Pi9基因序列标记的开发及其抗瘟育种应用,作物研究,28(3):231-235)
Yin D.S.,Xia M.Y.,Li J.B.,Wan B.L.,Zha Z.P.,Du X.S.,and Qi H.X.,2011,Development of STS marker linked to rice blast resistance gene Pi9 in marker-assisted selection breeding,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),25(1):25-30(殷得所,夏明元,李进波,万丙良,査中萍,杜雪树,戚华雄,2011,抗稻瘟病基因Pi9的STS连锁标记开发及在分子标记辅助育种中的应用,中国水稻科学,25(1):25-30)
Zou J.F.,Li Y.C.,Liu X.L.,Liu J.L.,Chen H.L.,Yang F.Y.,Huang J.,and Liao H.,2017,Improving blast resistance of rice restorer'E32'and its hybrid through molecular marker-assisted selection,Zuowu Yanjiu(Crop Research),31(1):11-14,24(邹俊锋,李永聪,刘雄伦,刘金灵,陈海龙,杨丰宇,黄俊,廖花,2017,MAS育种改良籼稻恢复系‘E32’及其杂交种的稻瘟病抗性,作物研究,31(1):11-14)
Bai B.,Wu J.,Zhou B.,and Deng Q.Y.,2012,Research progress of molecular breeding of rice blast resistance,Zajiao Shuidao(Hybrid Rice),27(3):5-9(柏斌,吴俊,周波,邓启云,2012,稻瘟病抗性分子育种研究综述,杂交水稻,27(3):5-9)
Bonman J.M.,VergeldeDios T.L.,and Khin M.M.,1986,Physiologic specialization of Pyricularia oryzae in the Philippines,Plant Disease,70(8):767-769
Cao Z.,Zeng G.,Hao M.,Sheng H.W.,Ye N.Z.,and Xiao Y.H.,2015,Improving blast resistance of dual-purpose genic sterile line C815S by using molecular marker-assisted selection,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(6):1193-1200(曹志,曾盖,郝明,盛浩闻,叶乃忠,肖应辉,2015,利用MAS技术改良水稻两用核不育系C815S的稻瘟病抗性,分子植物育种,13(6):1193-1200)
Chen H.L.,Li Y.C.,Liu X.L.,Liu J.L.,Yang F.Y.,Xing X.,Liao H.,Huang J.,and Liu J.F.,2017,Improving Blast Resistance of indica rice restorer R599 and its two-line hybrid using MAS technology,Zajiao Shuidao(Hybrid Rice),32(5):61-65(陈海龙,李永聪,刘雄伦,刘金灵,杨丰宇,行璇,廖花,黄俊,刘建丰,2017,MAS育种改良籼稻恢复系R599及其杂交种的稻瘟病抗性,杂交水稻,32(5):61-65)
Li Y.,Nie Y.Y.,Cai Y.H.,Yan M.L.,Mao L.H.,Li Y.H.,Lu M.,and Li M.D.,2011,Research progress in rice blast resistant genes in rice and their molecular marker assisted selection breeding,Jiangxi Nongye Xuebao(Acta Agriculturae Jiangxi),23(8):115-117(李瑶,聂元元,蔡耀辉,颜满莲,毛凌华,李永辉,卢明,李名迪,2011,水稻稻瘟病抗性基因及其分子标记辅助选择育种研究进展,江西农业学报,23(8):115-117)
Liu G.,Lu G.,Zeng L.,and Wang G.L.,2002,Two broad-spectrum blast resistance genes,Pi9(t)and Pi2(t),are physically linked on rice chromosome 6,Mol.Genet.Genomics,267(4):472-480
LüX.L.,Bai H.B.,Hui J.,Cai Z.Y.,Zhang L.F.,and Li S.H.,2017,The improvement of rice blast resistance by broadspectrum gene Pigm,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(11):4336-4342(吕学莲,白海波,惠建,蔡正云,张龙飞,李树华,2017,利用广谱抗病基因Pigm改良水稻稻瘟病抗性的研究,分子植物育种,15(11):4336-4342)
Moffat A.S.,1994,Plant genetics,mapping the sequence of disease resistance,Science,265(5180):1804-1805
Murray M.G.,and Thompson W.F.,1980,Rapid isolation of high molecular weight plant DNA,Nucleic Acids Res.,8(19):4321-4326
Ni D.H.,Yi C.X.,Li L.,Wang X.F.,Wang W.X.,and Yang J.B.,2005,Pyramiding Xa21 and Pi9(t)in rice by marker-assisted selection,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),3(3):329-334(倪大虎,易成新,李莉,汪秀峰,王文相,杨剑波,2005,利用分子标记辅助选择聚合水稻基因Xa21和Pi9(t),分子植物育种,3(3):329-334)
Qu S.H.,Liu G.F.,Zhou B.,Bellizzi M.,Zeng L.R.,Dai L.Y.,Han B.,and Wang G.L.,2006,The broad-spectrum blast resistance gene Pi9 encodes a nucleotide-binding site-leucinerich repeat protein and is a member of a multigene family in rice,Genetics,172(3):1901-1914
Tan L.Q.,Liu X.L.,Yang T.T.,Xing X.,Liang Y.,Chen H.L.,Yang F.Y.,Li Y.C.,Liu J.L.,Dai L.Y.,Chen L.Y.,and Wang G.L.,2015,Breeding new rice line with blast resistance by molecular marker-assisted selection of the Pi9gene,Zuowu Yanjiu(Crop Research),29(4):348-351,356(谭令辞,刘雄伦,杨婷婷,行璇,梁毅,陈海龙,杨丰宇,李永聪,刘金灵,戴良英,陈立云,王国梁,2015,利用Pi9基因分子标记辅助选择培育抗稻瘟病水稻新品系,作物研究,29(4):348-351,356)
Xiao G.,Borja F.N.,Mauleon R.,Padilla J.,Telebanco-Yanoria M.J.,Yang J.,Lu G.D.,Dionisio-Sese M.,and Zhou Bo.,2017,Identification of resistant germplasm containing novel resistance genes at or tightly linked to the Pi2/9 locus conferring broad-spectrum resistance against rice blast,Rice,10(1):37
Xing X.,Liu X.L.,Chen H.L.,Yang F.Y.,Li Y.C.,Liao H.,You L.,Liu J.L.,Dai L.Y.,and Wang G.L.,2016,Improving blast resistance of rice restorer R288 by molecular marker-assisted selection of Pi9 gene,Zuowu Yanjiu(Crop Research),30(5):487-491(行璇,刘雄伦,陈海龙,杨丰宇,李永聪,廖花,游亮,刘金灵,戴良英,王国梁,2016,分子标记辅助选择Pi9基因改良R288的稻瘟病抗性,作物研究,30(5):487-491)
Yang F.Y.,Li Y.C.,Liu X.L.,Liu J.L.,Chen H.L.,Liao H.,Huang J.,and Liang Y.,2017,Improving blast resistance of the early indica rice 1701 by molecular marker-assisted selection,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(6):2212-2217(杨丰宇,李永聪,刘雄伦,刘金灵,陈海龙,廖花,黄俊,梁毅,2017,分子标记辅助选择改良早籼稻1701的稻瘟病抗性,分子植物育种,15(6):2212-2217)
Yang T.T.,Liu X.L.,Tan L.Q.,Xing X.,Wen T.,Liang Y.,Wu J.,Liu J.L.,Dai L.Y.,Liu J.F.,and Wang G.L.,2014,Development of the Pi9 gene marker and its breeding practice in rice blast resistance,Zuowu Yanjiu(Crop Research),28(3):231-235(杨婷婷,刘雄伦,谭令辞,行璇,文婷,梁毅,吴俊,刘金灵,戴良英,刘建丰,王国梁,2014,水稻Pi9基因序列标记的开发及其抗瘟育种应用,作物研究,28(3):231-235)
Yin D.S.,Xia M.Y.,Li J.B.,Wan B.L.,Zha Z.P.,Du X.S.,and Qi H.X.,2011,Development of STS marker linked to rice blast resistance gene Pi9 in marker-assisted selection breeding,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),25(1):25-30(殷得所,夏明元,李进波,万丙良,査中萍,杜雪树,戚华雄,2011,抗稻瘟病基因Pi9的STS连锁标记开发及在分子标记辅助育种中的应用,中国水稻科学,25(1):25-30)
Zou J.F.,Li Y.C.,Liu X.L.,Liu J.L.,Chen H.L.,Yang F.Y.,Huang J.,and Liao H.,2017,Improving blast resistance of rice restorer'E32'and its hybrid through molecular marker-assisted selection,Zuowu Yanjiu(Crop Research),31(1):11-14,24(邹俊锋,李永聪,刘雄伦,刘金灵,陈海龙,杨丰宇,黄俊,廖花,2017,MAS育种改良籼稻恢复系‘E32’及其杂交种的稻瘟病抗性,作物研究,31(1):11-14)