利用Pi-k~h基因改良恢复系稻瘟病抗性
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摘要
为改良新选育的两个恢复系R1、R2的稻瘟病抗性,挑选出含有抗稻瘟病基因的改良后代,本研究通过分子标记技术,利用携带广谱抗稻瘟病基因Pi-kh的Tetep为供体,R1,R2为受体亲本,对其杂交后代进行选择。实验结果显示分子标记RM224在亲本间有良好的多态性,在稻瘟病抗性选择中的正确率较高,大大缩短了育种年限。实验选育的两个改良恢复系在田间鉴定中有较高的稻瘟病抗性,在与不同不育系配制的组合中也存在较高的稻瘟病抗性和在产量上的超标优势,平均优势率达5%,说明改良的恢复系在实际生产上有良好的生产潜力和应用前景。
In order to improve the rice blast resistance of the newly-selected two restorer lines R1 and R2 and select improved progeny containing blast resistance genes, Tetep, a donor carrying a broad spectrum rice blast resistance gene Pi-kh, was used as a donor, R1 and R2 were used as receptor parents, and their hybrids were selected by molecular marker technology in this study. The experimental results showed that the molecular marker RM224 had good polymorphism among the parents, and the correct rate in selection was higher, which greatly shortened the breeding periods. The two improved restorer lines selected had higher resistance to rice blast in the field identification. There was also high rice blast resistance and excessive yield advantage in the combinations with different sterile lines. The average dominance rate was 5%, indicating that the improved restorer line had a good application prospect and potential in production.
In order to improve the rice blast resistance of the newly-selected two restorer lines R1 and R2 and select improved progeny containing blast resistance genes, Tetep, a donor carrying a broad spectrum rice blast resistance gene Pi-kh, was used as a donor, R1 and R2 were used as receptor parents, and their hybrids were selected by molecular marker technology in this study. The experimental results showed that the molecular marker RM224 had good polymorphism among the parents, and the correct rate in selection was higher, which greatly shortened the breeding periods. The two improved restorer lines selected had higher resistance to rice blast in the field identification. There was also high rice blast resistance and excessive yield advantage in the combinations with different sterile lines. The average dominance rate was 5%, indicating that the improved restorer line had a good application prospect and potential in production.
引文
Bai Z.A.,Ren Z.H.,Mao Y.,Zhou L.,Li S.,and Liu E.M.,2011,Analysis of genetic diversity of Maganaporthe oryzae in rice blast nurseries in Taojiang and Liuyang by SSR markers,Hunan Nongye Kexue(Hunan Agricultural Sciences),(7):7-10(白珍安,任佐华,毛莹,周翎,李书,刘二明,2011,桃江和浏阳病圃稻瘟病菌遗传多样性的SSR分析,湖南农业科学,(7):7-10)
Chen B.T.,Peng Z.M.,and Xu Y.Q.,1993,Inheritance of blastresistance and application of blast-resistance variety Tetep,Zhiwu Baohu Xuebao(Journal of Plant Protection),20(3):233-236(陈葆棠,彭仲明,徐运启,1993,稻瘟病抗源Tetep的抗性遗传及其应用,植物保护学报,20(3):233-236)
Chen Z.W.,Guan H.Z.,Wang Z.H.,Zhou Y.C.,Pan R.S.,Mao D.M.,Duan Y.L.,and Wu W.R.,2012,Improvement of the resistance to rice blast in a high quality rice CMS line by gene pyramiding,Fujian Nonglin Daxue Xuebao(Journal of Fujian Agriculture and Forestry University(Natural Science Edition)),41(5):449-454(陈志伟,官华忠,王宗华,周元昌,潘润森,毛大梅,段远霖,吴为人,2012,利用基因聚合技术改良一个优质水稻不育系的稻瘟病抗性,福建农林大学学报(自然科学版),41(5):449-454)
Chen S.,Lin X.H.,Xu C.G.,and Zhang Q.F.,2000,Improvement of bacterial blight resistance of'Minghui 63',an elite restorer line of hybrid rice,by molecular marker-assisted se lection,Crop Sci.,(40):239-244
Chen X.,Temnykh S.,Xu Y.,Cho Y.G.,and Mccouch S.R.,1997,Development of a microsatellite framework map providing genome-wide coverage in rice(Oryza sativa L.),Theor.Appl.Genet.,95(4):553-567
Jiang Y.L.,2008,Studies on breeding of parents of hybrid rice with durable disease resistance using molecular marker-assisted seleetion,Thesis for M.S.,Agriculture,Fujian Agriculture and Forestry University,Supervisor:Wu W.R.,pp8-71(蒋云林,2008,利用MAS技术选育持久抗病杂交水稻亲本的研究,硕士学位论文,福建农林大学农学院,导师:吴为人,pp.8-71)
Liang Y.,Tang T.T.,Tan L.C.,Wen T.,Wu J.,Jiang N.,Li Z.Q.Dai L.Y.,Wang G.L.,and Liu X.L.,2013,Development of the molecular marker tightly-linked with the broad-spectrum blast resistance gene Pigm and its breeding practice in rice Zajiao Shuidao(Hybrid Rice),28(4):63-68(梁毅,杨婷婷谭令辞,文婷,吴俊,江南,李智强,戴良英,王国梁,刘雄伦,2013,水稻广谱抗瘟基因Pigm紧密连锁分子标记开发及其育种应用,杂交水稻,28(4):63-68)
Lei C.L.,Wang J.L.,Jiang W.R.,Ling Z.Z.,and George M.L.2002,Population structure and genetic variation of rice blast fungus in some rice-growing regions in northern China,Zhiwu Bingli Xuebao(Acta Phytopathologica Sinica)32(3):219-226(雷财林,王久林,蒋琬如,凌忠专,George M.L.,2002,我国北方部分稻区稻瘟病菌群体遗传结构研究,植物病理学报,32(3):219-226)
Sharma T.R.,Madhav M.S.,Singh B.K.,Shanker P.,Jana T.K.Dalal V.,Pandit A.,Singh A.,Gaikwad K.,Upreti H.C.,and Singh N.K.,2005,High-resolution mapping,cloning and molecular characterization of the Pi-khgene of rice,which confers resistance to Magnaporthe grisea,Mol.Gen.Genomics.,274(6):569-578
Song G.S.,Sun L.,Liu Z.Q.,Chen M.J.,Zhu X.X.,Zhou G.C.Feng Y.,Pu G.J.,and Hou L.G.,2016 Rice blast resistance analysis and molecular assistant improvement of main rice varieties in Jilin Province,Xibei Nongye Xuebao(Acta A-griculturae Boreali-occidentalis Sinica),25(5):677-683(宋广树,孙蕾,刘志全,陈寞军,朱秀侠,周广春,封云,浦桂君,侯立刚,2016,吉林省主要水稻品种稻瘟病抗性分析及分子辅助改良,西北农业学报,25(5):677-683)
Wang J.,Yang J.,Zhu J.Y.,Fan F.J.,Li W.Q.,Wang F.Q.,Huang Z.Y.,and Zhong W.G.,2014,Development of a functiona marker for rice blast resistance gene Pi-khand natural variation at Pi-khlocus in japonica rice in Jiangsu Province,C-hina,Zhongguo Shuidao Kexue(Chinese Journal of Rice S-cience),28(2):141-147(王军,杨杰,朱金燕,范方军,李文奇,王芳权,黄转运,仲维功,2014,稻瘟病抗病基因Pi-kh功能标记的开发及江苏粳稻品种中Pi-kh的变异,中国水稻科学,28(2):141-147)
Wu H.,2013,Pyramiding Pik-h and Pi-ta gene in the rice maintainer line BoⅢB(Oryza satita L.)and the isolation of Pyricutaria oryzae fungus,Thesis for M.S.,Guangxi University,Supervisor:Liu P.Q.,pp.5-48(伍豪,2013,在优质水稻保持系博ⅢB中聚合抗稻瘟病基因Pik-h和Pi-ta以及稻瘟病菌的分离,硕士学位论文,广西大学,导师:刘丕庆,pp.5-48)
Wen T.,Liang Y.,Jiang N.,Li Z.Q.,Liu J.L.,Yang T.T.,Dai L.Y.,Wang G.L.,and Liu X.L.,2012,Improving blast resistance of indica rice lines by Pi9 gene marker-assisted selection,Hunan Nongye Daxue Xuebao(Journal of Hunan Agricultural University(Natural Sciences)),38(3):262-266(文婷,梁毅,江南,李智强,刘金灵,杨婷婷,戴良英,王国梁,刘雄伦,2012,利用Pi9基因序列标记辅助选择改良籼稻稻瘟病抗性,湖南农业大学学报(自然科学版),38(3):262-266)
Yang T.T.,Liu X.L.,Tan L.C.,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)
Yang J.Y.,Chen S.,Zeng L.X.,Li Y.L.,Chen Z.,and Zhu X.Y.,2008,Evaluation on resistance of major rice blast resistance genes to Magnaporthe grisea isolates collected from indica rice in Guangdong Province,China,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),22(2):190-196(杨健源,陈深,曾列先,李亦龙,陈珍,朱小源,2008,稻瘟病主效抗性基因对广东省籼稻稻瘟病菌的抗性评价,中国水稻科学,22(2):190-196)
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):11-14,24)
Chen B.T.,Peng Z.M.,and Xu Y.Q.,1993,Inheritance of blastresistance and application of blast-resistance variety Tetep,Zhiwu Baohu Xuebao(Journal of Plant Protection),20(3):233-236(陈葆棠,彭仲明,徐运启,1993,稻瘟病抗源Tetep的抗性遗传及其应用,植物保护学报,20(3):233-236)
Chen Z.W.,Guan H.Z.,Wang Z.H.,Zhou Y.C.,Pan R.S.,Mao D.M.,Duan Y.L.,and Wu W.R.,2012,Improvement of the resistance to rice blast in a high quality rice CMS line by gene pyramiding,Fujian Nonglin Daxue Xuebao(Journal of Fujian Agriculture and Forestry University(Natural Science Edition)),41(5):449-454(陈志伟,官华忠,王宗华,周元昌,潘润森,毛大梅,段远霖,吴为人,2012,利用基因聚合技术改良一个优质水稻不育系的稻瘟病抗性,福建农林大学学报(自然科学版),41(5):449-454)
Chen S.,Lin X.H.,Xu C.G.,and Zhang Q.F.,2000,Improvement of bacterial blight resistance of'Minghui 63',an elite restorer line of hybrid rice,by molecular marker-assisted se lection,Crop Sci.,(40):239-244
Chen X.,Temnykh S.,Xu Y.,Cho Y.G.,and Mccouch S.R.,1997,Development of a microsatellite framework map providing genome-wide coverage in rice(Oryza sativa L.),Theor.Appl.Genet.,95(4):553-567
Jiang Y.L.,2008,Studies on breeding of parents of hybrid rice with durable disease resistance using molecular marker-assisted seleetion,Thesis for M.S.,Agriculture,Fujian Agriculture and Forestry University,Supervisor:Wu W.R.,pp8-71(蒋云林,2008,利用MAS技术选育持久抗病杂交水稻亲本的研究,硕士学位论文,福建农林大学农学院,导师:吴为人,pp.8-71)
Liang Y.,Tang T.T.,Tan L.C.,Wen T.,Wu J.,Jiang N.,Li Z.Q.Dai L.Y.,Wang G.L.,and Liu X.L.,2013,Development of the molecular marker tightly-linked with the broad-spectrum blast resistance gene Pigm and its breeding practice in rice Zajiao Shuidao(Hybrid Rice),28(4):63-68(梁毅,杨婷婷谭令辞,文婷,吴俊,江南,李智强,戴良英,王国梁,刘雄伦,2013,水稻广谱抗瘟基因Pigm紧密连锁分子标记开发及其育种应用,杂交水稻,28(4):63-68)
Lei C.L.,Wang J.L.,Jiang W.R.,Ling Z.Z.,and George M.L.2002,Population structure and genetic variation of rice blast fungus in some rice-growing regions in northern China,Zhiwu Bingli Xuebao(Acta Phytopathologica Sinica)32(3):219-226(雷财林,王久林,蒋琬如,凌忠专,George M.L.,2002,我国北方部分稻区稻瘟病菌群体遗传结构研究,植物病理学报,32(3):219-226)
Sharma T.R.,Madhav M.S.,Singh B.K.,Shanker P.,Jana T.K.Dalal V.,Pandit A.,Singh A.,Gaikwad K.,Upreti H.C.,and Singh N.K.,2005,High-resolution mapping,cloning and molecular characterization of the Pi-khgene of rice,which confers resistance to Magnaporthe grisea,Mol.Gen.Genomics.,274(6):569-578
Song G.S.,Sun L.,Liu Z.Q.,Chen M.J.,Zhu X.X.,Zhou G.C.Feng Y.,Pu G.J.,and Hou L.G.,2016 Rice blast resistance analysis and molecular assistant improvement of main rice varieties in Jilin Province,Xibei Nongye Xuebao(Acta A-griculturae Boreali-occidentalis Sinica),25(5):677-683(宋广树,孙蕾,刘志全,陈寞军,朱秀侠,周广春,封云,浦桂君,侯立刚,2016,吉林省主要水稻品种稻瘟病抗性分析及分子辅助改良,西北农业学报,25(5):677-683)
Wang J.,Yang J.,Zhu J.Y.,Fan F.J.,Li W.Q.,Wang F.Q.,Huang Z.Y.,and Zhong W.G.,2014,Development of a functiona marker for rice blast resistance gene Pi-khand natural variation at Pi-khlocus in japonica rice in Jiangsu Province,C-hina,Zhongguo Shuidao Kexue(Chinese Journal of Rice S-cience),28(2):141-147(王军,杨杰,朱金燕,范方军,李文奇,王芳权,黄转运,仲维功,2014,稻瘟病抗病基因Pi-kh功能标记的开发及江苏粳稻品种中Pi-kh的变异,中国水稻科学,28(2):141-147)
Wu H.,2013,Pyramiding Pik-h and Pi-ta gene in the rice maintainer line BoⅢB(Oryza satita L.)and the isolation of Pyricutaria oryzae fungus,Thesis for M.S.,Guangxi University,Supervisor:Liu P.Q.,pp.5-48(伍豪,2013,在优质水稻保持系博ⅢB中聚合抗稻瘟病基因Pik-h和Pi-ta以及稻瘟病菌的分离,硕士学位论文,广西大学,导师:刘丕庆,pp.5-48)
Wen T.,Liang Y.,Jiang N.,Li Z.Q.,Liu J.L.,Yang T.T.,Dai L.Y.,Wang G.L.,and Liu X.L.,2012,Improving blast resistance of indica rice lines by Pi9 gene marker-assisted selection,Hunan Nongye Daxue Xuebao(Journal of Hunan Agricultural University(Natural Sciences)),38(3):262-266(文婷,梁毅,江南,李智强,刘金灵,杨婷婷,戴良英,王国梁,刘雄伦,2012,利用Pi9基因序列标记辅助选择改良籼稻稻瘟病抗性,湖南农业大学学报(自然科学版),38(3):262-266)
Yang T.T.,Liu X.L.,Tan L.C.,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)
Yang J.Y.,Chen S.,Zeng L.X.,Li Y.L.,Chen Z.,and Zhu X.Y.,2008,Evaluation on resistance of major rice blast resistance genes to Magnaporthe grisea isolates collected from indica rice in Guangdong Province,China,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),22(2):190-196(杨健源,陈深,曾列先,李亦龙,陈珍,朱小源,2008,稻瘟病主效抗性基因对广东省籼稻稻瘟病菌的抗性评价,中国水稻科学,22(2):190-196)
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):11-14,24)