5个稻瘟病抗性基因在云南水稻育种中的利用分析
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摘要
为明确Pi9等5个抗稻瘟病主效基因在云南省抗稻瘟病育种中的利用状况,明确种质资源携带的抗性基因,本研究用4个分子标记对云南省80年代以来育成的60个推广品种和23份优异地方稻种的Pik-h、Pi2、Pi9、Pi5抗稻瘟病基因进行了分子检测,并对抗稻瘟病基因Pita第二编码区功能位点碱基进行了测序分析。结果表明:Pik-h、Pi5、Pita在地方品种中出现频率比较高,分别为65.22%、52.17%和43.48%,而Pi9和Pi2极少或未出现;现代育成推广品种中Pik-h、Pi5、Pi9、Pita频率分别为70%,38.33%,33.33%和11.67%,仅1个品种携带Pi2基因;一个品种一般携带5个基因中的0~3个;根据基因组合,可将83个品种归为14种基因型,现代育成品种具有14种,其中Pik-h、(Pik-h)-Pi5、(Pik-h)-Pi9等组合是优势类型;地方品种具有14种基因组合中的9种,2000年后审定品种中不携带或只携带1个基因的品种比例有所增加,且集中在Pik-h,比例高达此类品种的57.89%,抗性基因单一化严重,易造成成批品种抗性丧失。
In order to clarify the utilization status of five major resistant genes to rice blast in rice breeding in Yunnan province and to identify the resistance genes of germplasm resources, we used four molecular markers to detect the rice blast resistant genes Pik-h, Pi2, Pi9 and Pi5 of 60 promotion varieties and 23 excellent local rice varieties from Yunnan province since the 1980 s, and analyzed the sequences of functional site base of the second coding region in the resistance gene Pita. The results showed that Pik-h, Pi5 and Pita showed high occurrence frequencies in local varieties, which were 65.22%, 52.17% and 43.48% respectively, while Pi9 and Pi2 hardly occurred. In the modern varieties, the frequencies of Pik-h, Pi5, Pi9 and Pita were 70%, 38.33%, 33.33% and11.67%, respectively. Only one cultivar carried Pi2 gene, and a variety usually carried 0~3 of the 5 genes.According to the genes combinations, 83 varieties could be classified into 14 genotypes and modern cultivars had14 kinds, among which Pik-h,(Pik-h)-Pi5,(Pik-h)-Pi9 were the dominant types. Local varieties had 9 of 14 genot-ypes. Since 2000 years, the proportion of detected cultivars that had no gene or only one gene has increased, and the rate of cultivars with single gene Pik-h was as high as 57.89% among these cultivars with single or noncarrying gene. The resistance gene was seriously simple and quite susceptible to the loss of resistance in batches.
In order to clarify the utilization status of five major resistant genes to rice blast in rice breeding in Yunnan province and to identify the resistance genes of germplasm resources, we used four molecular markers to detect the rice blast resistant genes Pik-h, Pi2, Pi9 and Pi5 of 60 promotion varieties and 23 excellent local rice varieties from Yunnan province since the 1980 s, and analyzed the sequences of functional site base of the second coding region in the resistance gene Pita. The results showed that Pik-h, Pi5 and Pita showed high occurrence frequencies in local varieties, which were 65.22%, 52.17% and 43.48% respectively, while Pi9 and Pi2 hardly occurred. In the modern varieties, the frequencies of Pik-h, Pi5, Pi9 and Pita were 70%, 38.33%, 33.33% and11.67%, respectively. Only one cultivar carried Pi2 gene, and a variety usually carried 0~3 of the 5 genes.According to the genes combinations, 83 varieties could be classified into 14 genotypes and modern cultivars had14 kinds, among which Pik-h,(Pik-h)-Pi5,(Pik-h)-Pi9 were the dominant types. Local varieties had 9 of 14 genot-ypes. Since 2000 years, the proportion of detected cultivars that had no gene or only one gene has increased, and the rate of cultivars with single gene Pik-h was as high as 57.89% among these cultivars with single or noncarrying gene. The resistance gene was seriously simple and quite susceptible to the loss of resistance in batches.
引文
Bryan G.T.,Wu K.S.,Farrall L.,Jia Y.,Hershey H.P.,Mc Adams S.A.,Faulk K.N.,Donaldson G.K.,Tarchini R.,and Valen B.,2000,TA single amino acid difference distinguishes resistant and susceptible alleles of the rice blast resistance gene Pi-ta,Plant Cell,12(11):2033-2046
Chen H.L.,Chen B.T.,Zhang D.P.,Xie Y.F.,and Zhang Q.F.2001,Pathotypes of Pyricularia grisea in Rice Fields of central and southern China,Plant Disease,85(8):843-850
Dai X.J.,Yang Y.Z.,Zhou L.,Liang M.Z.,Hu X.C.,and Chen LB.,2012,Distribution research of blast resistance genes pita pib,pi9 and pikm in blast-resistant rice resources,Shengming Kexue Yanjiu(Life Science Research),16(4):340-344(戴小军,杨远柱,周亮,梁满中,胡小淳,陈良碧,2012,抗稻瘟病水稻资源抗性基因Pita,Pib,Pi9以及Pikm的分布研究,生命科学研究,16(4):340-344)
He C.,Chen T.,Zhang Y.D.,Zhu Z.,Zhao Q.Y.,Zhou L.H.,Yu X.,and Wang C.L.,2014,Genotypic analysis of blast resistance genes Pi-ta and Pi-b for japonica rice varieties and lines in Jiangsu Province,Jiangsu Nongye Xuebao(Jiangsu Journal of Agricultural Sciences),30(5):921-927(何重,陈涛,张亚东,朱镇,赵庆勇,周丽慧,于新,王才林,2014,江苏部分粳稻品种和品系中稻瘟病抗性基因Pi-ta和Pi-b的基因型分析,江苏农业学报,30(5):921-927)
Jin G.G.,Zhao Y.D.,Jin J.H.,Piao R.H.,Wang J.M.,and Zhang Q.,2017,Molecular marker detection of rice blast resistance gene from 196 rice varieties in Jilin Province,Beifang Shuidao(North Rice),47(2):5-12(金国光,赵亚东,金京花,朴日花,王金明,张强,2017,196份吉林省水稻材料的稻瘟病抗性基因分子标记检测,北方水稻,47(2):5-12)
Lee S.K.,Song M.Y.,Seo Y.S.,Kim H.K.,Ko S.H.,Cao P.,Suh J.P.,Yi G.,Roh J.H.,Lee S.,An G.,Hahn T.R.,Wang G.L.,Ronald P.,and Jeon J.S.,2009,Rice Pi5-Mediated resistance to Magnaporthe oryzae requires the presence of two oiled-coil-nucleotide-binding-leucine-rich,Repeat Genes Genetics,181(4):1627-1638
Li J.B.,Li C.Y.,Chen Y.,Lei C.L.,and Lin Z.Z.,2005,Evaluation of twenty-two blast resistance genes in Yunnan using monogenetic rice lines,Zhiwu Baohu Xuebao(Acta Phytophylacica Sinica),32(2):113-119(李进斌,李成云,陈艳,雷财林,凌忠专,2005,二十二个抗稻瘟病基因在云南的利用价值评价,植物保护学报,32(2):113-119)
Liu G.,Lu G.,Zeng L.,and Wang G.L.,2002,Two broadspectrum blast resistance genes,Pi9(t)and Pi2(t),are physically linked on rice chromosome 6,Mol.Genet.Genomics,267(4):472-480
Ma J.Q.,Yang Y.,Sun Y.D.,Wang Y.Y.,and Xu M.H.,2016,Sequence variation of functional fragment in Pitagene for rice blast resistance i n Yunnan rice landraces,Zhongguo Shuidao Kexue(Chinses Journal of Rice Science),30(3):265-272(马继琼,杨奕,孙一丁,王炎炎,许明辉,2016,云南地方水稻品种抗稻瘟病基因Pita功能区段序列变异分析,中国水稻科学,30(3):265-272)
Ma J.T.,Zhang G.M.,Xin A.H.,Zhang L.Y.,Deng L.W.,and Ren Y.,2016,Resistance analysis and improvement of rice varieties by gene pyramiding,Zhiwu Baohu Xuebao(Journal of Plant Protection),43(2):177-183(马军韬,张国民,辛爱华,张丽艳,邓凌韦,任洋,2016,水稻品种抗稻瘟病分析及基因聚合抗性改良,植物保护学报,43(2):177-183)
Shi K.,Lei C.L.,Cheng Z.J.,Xu X.T.,Wang J.L.,and Wan J.M.,2009,Distribution of two blast resistance genes pita and pib in major rice cultivars in China,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),10(1):21-26(时克,雷财林,程治军,许兴涛,王久林,万建明,2009,稻瘟病抗性基因Pita和Pib在我国水稻主栽品种中的分布植物遗传资源学报,10(1):21-26)
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 functional marker for rice blast resitance gene Pi-kh and natura variationg at Pi-kh locus in japonica rice in Jiangsu Province Zhongguo Shuidao Kexue(Chinese Journal of Rice Science)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)
Wang S.X.,Li J.,Zhou X.Y.,Li Y.,Li M.Q.,Cao B.W.,and Zhao Q.Z.,2017,Molecular detection of rice blast resistance Gene Pi9,Pita and Piz-t in Henan Japonica Rice,Fenz Zhiwu Yuzhong(Molecular Plant Breeding),15(3):951-955(王生轩,李俊,周谢瑛,李勇,李梦琪,曹炳武,赵全志2017,河南粳稻抗稻瘟病基因Pi9,Pita和Piz-t的分子检测,分子植物育种,15(3):951-955)
Wang W.J.,Zhou J.Y.,Wang C.Y.,Su J.,Feng J.Q.,Chen B.Feng A.Q.,Yang J.Y.,Chen S.,and Zhu X.Y.,2017,Distribution of eight rice blast resistance genes in indica hybrid rice in China,Zhongguo Shuidao Kexue(Chinses Journal of Rice Science),31(3):299-306(汪文娟,周继勇,汪聪颖,苏菁,封金奇,陈炳,冯爱卿,杨健源,陈深,朱小源,2017,八个抗稻瘟病基因在华南籼型杂交水稻中的分布,中国水稻科学,31(3):299-306)
Wang Y.Y.,Sun Y.D.,Ma J.Q.,Yang Y.,and Xu M.H.,2016Preliminary study on the characteristics SNP of microsatellite sequence(CT)n and intron 1 in Wx gene in wild rices,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(12):3458-3464(王炎炎,孙一丁,马继琼,杨奕,许明辉,2016,野生稻Wx基因(CT)n卫星序列和第一内含子SNP特征标签初探,基因组学与应用生物学,35(12):3458-3464)
Yan Y.,Ma Z.F.,Qin G.,Chen Y.M.,Qin Y.Y.,Yan Q.,Liu C.,Zhang Y.X.,and Huang D.H.,2017,Evaluation of blast resistance and identification of resistance genes in main indica rice patent materials in South China,Nanfang Nongye Xuebao(Journal of Southern Agriculture),48(4):587-593(阎勇,马增凤,秦钢,陈远孟,秦媛媛,颜群,刘驰,张月雄,黄大辉,2017,华南常用籼稻亲本稻瘟病抗性评价及抗性基因鉴定,南方农业学报,48(4):587-593)
Zhang Y.,Feng Z.F.,Cui M.Z.,and Zhang X.J.,2013a,Distribution of blast resistant gene Pi-km in rice germplasm in Shanxi province of northwest China,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),11(3):311-316(张羽,冯志峰,崔明珠,张晓娟,2013a,稻瘟病抗性基因Pi-km在陕西省稻种资源中的分布状况,分子植物育种,11(3):311-316)
Zhang Y.,Feng Z.F.,Zhang H.,and Mo D.,2013b,Distribution of blast resistant gene Pi9 in rice germplasm in Shanxi Province,Sichuan Nongye Daxue Xuebao(Journal of Sichuan Agricultural University),31(2):115-121(张羽,冯志峰,张晗,莫丹,2013b,陕西省水稻种质资源中Pi9基因的分布状况,四川农业大学学报,31(2):115-121)
Zou D.T.,Jiang S.D.,Zhao H.W.,Guo L.Y.,Sun J.,Liu H.L.,and Xin W.,2016,Distribution of broad-spectrum resistance gene-Pi9 in the rice cultivars of Heilongjiang Province,Dongbei Nongye Daxue Xuebao(Journal of Northeast Agricultural University),47(7):1-8(邹德堂,姜思达,赵宏伟,郭丽颖,孙健,刘化龙,辛威,2016,广谱抗性基因Pi9在黑龙江省水稻品种中的分布,东北农业大学学报,47(7):1-8)
Chen H.L.,Chen B.T.,Zhang D.P.,Xie Y.F.,and Zhang Q.F.2001,Pathotypes of Pyricularia grisea in Rice Fields of central and southern China,Plant Disease,85(8):843-850
Dai X.J.,Yang Y.Z.,Zhou L.,Liang M.Z.,Hu X.C.,and Chen LB.,2012,Distribution research of blast resistance genes pita pib,pi9 and pikm in blast-resistant rice resources,Shengming Kexue Yanjiu(Life Science Research),16(4):340-344(戴小军,杨远柱,周亮,梁满中,胡小淳,陈良碧,2012,抗稻瘟病水稻资源抗性基因Pita,Pib,Pi9以及Pikm的分布研究,生命科学研究,16(4):340-344)
He C.,Chen T.,Zhang Y.D.,Zhu Z.,Zhao Q.Y.,Zhou L.H.,Yu X.,and Wang C.L.,2014,Genotypic analysis of blast resistance genes Pi-ta and Pi-b for japonica rice varieties and lines in Jiangsu Province,Jiangsu Nongye Xuebao(Jiangsu Journal of Agricultural Sciences),30(5):921-927(何重,陈涛,张亚东,朱镇,赵庆勇,周丽慧,于新,王才林,2014,江苏部分粳稻品种和品系中稻瘟病抗性基因Pi-ta和Pi-b的基因型分析,江苏农业学报,30(5):921-927)
Jin G.G.,Zhao Y.D.,Jin J.H.,Piao R.H.,Wang J.M.,and Zhang Q.,2017,Molecular marker detection of rice blast resistance gene from 196 rice varieties in Jilin Province,Beifang Shuidao(North Rice),47(2):5-12(金国光,赵亚东,金京花,朴日花,王金明,张强,2017,196份吉林省水稻材料的稻瘟病抗性基因分子标记检测,北方水稻,47(2):5-12)
Lee S.K.,Song M.Y.,Seo Y.S.,Kim H.K.,Ko S.H.,Cao P.,Suh J.P.,Yi G.,Roh J.H.,Lee S.,An G.,Hahn T.R.,Wang G.L.,Ronald P.,and Jeon J.S.,2009,Rice Pi5-Mediated resistance to Magnaporthe oryzae requires the presence of two oiled-coil-nucleotide-binding-leucine-rich,Repeat Genes Genetics,181(4):1627-1638
Li J.B.,Li C.Y.,Chen Y.,Lei C.L.,and Lin Z.Z.,2005,Evaluation of twenty-two blast resistance genes in Yunnan using monogenetic rice lines,Zhiwu Baohu Xuebao(Acta Phytophylacica Sinica),32(2):113-119(李进斌,李成云,陈艳,雷财林,凌忠专,2005,二十二个抗稻瘟病基因在云南的利用价值评价,植物保护学报,32(2):113-119)
Liu G.,Lu G.,Zeng L.,and Wang G.L.,2002,Two broadspectrum blast resistance genes,Pi9(t)and Pi2(t),are physically linked on rice chromosome 6,Mol.Genet.Genomics,267(4):472-480
Ma J.Q.,Yang Y.,Sun Y.D.,Wang Y.Y.,and Xu M.H.,2016,Sequence variation of functional fragment in Pitagene for rice blast resistance i n Yunnan rice landraces,Zhongguo Shuidao Kexue(Chinses Journal of Rice Science),30(3):265-272(马继琼,杨奕,孙一丁,王炎炎,许明辉,2016,云南地方水稻品种抗稻瘟病基因Pita功能区段序列变异分析,中国水稻科学,30(3):265-272)
Ma J.T.,Zhang G.M.,Xin A.H.,Zhang L.Y.,Deng L.W.,and Ren Y.,2016,Resistance analysis and improvement of rice varieties by gene pyramiding,Zhiwu Baohu Xuebao(Journal of Plant Protection),43(2):177-183(马军韬,张国民,辛爱华,张丽艳,邓凌韦,任洋,2016,水稻品种抗稻瘟病分析及基因聚合抗性改良,植物保护学报,43(2):177-183)
Shi K.,Lei C.L.,Cheng Z.J.,Xu X.T.,Wang J.L.,and Wan J.M.,2009,Distribution of two blast resistance genes pita and pib in major rice cultivars in China,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),10(1):21-26(时克,雷财林,程治军,许兴涛,王久林,万建明,2009,稻瘟病抗性基因Pita和Pib在我国水稻主栽品种中的分布植物遗传资源学报,10(1):21-26)
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 functional marker for rice blast resitance gene Pi-kh and natura variationg at Pi-kh locus in japonica rice in Jiangsu Province Zhongguo Shuidao Kexue(Chinese Journal of Rice Science)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)
Wang S.X.,Li J.,Zhou X.Y.,Li Y.,Li M.Q.,Cao B.W.,and Zhao Q.Z.,2017,Molecular detection of rice blast resistance Gene Pi9,Pita and Piz-t in Henan Japonica Rice,Fenz Zhiwu Yuzhong(Molecular Plant Breeding),15(3):951-955(王生轩,李俊,周谢瑛,李勇,李梦琪,曹炳武,赵全志2017,河南粳稻抗稻瘟病基因Pi9,Pita和Piz-t的分子检测,分子植物育种,15(3):951-955)
Wang W.J.,Zhou J.Y.,Wang C.Y.,Su J.,Feng J.Q.,Chen B.Feng A.Q.,Yang J.Y.,Chen S.,and Zhu X.Y.,2017,Distribution of eight rice blast resistance genes in indica hybrid rice in China,Zhongguo Shuidao Kexue(Chinses Journal of Rice Science),31(3):299-306(汪文娟,周继勇,汪聪颖,苏菁,封金奇,陈炳,冯爱卿,杨健源,陈深,朱小源,2017,八个抗稻瘟病基因在华南籼型杂交水稻中的分布,中国水稻科学,31(3):299-306)
Wang Y.Y.,Sun Y.D.,Ma J.Q.,Yang Y.,and Xu M.H.,2016Preliminary study on the characteristics SNP of microsatellite sequence(CT)n and intron 1 in Wx gene in wild rices,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(12):3458-3464(王炎炎,孙一丁,马继琼,杨奕,许明辉,2016,野生稻Wx基因(CT)n卫星序列和第一内含子SNP特征标签初探,基因组学与应用生物学,35(12):3458-3464)
Yan Y.,Ma Z.F.,Qin G.,Chen Y.M.,Qin Y.Y.,Yan Q.,Liu C.,Zhang Y.X.,and Huang D.H.,2017,Evaluation of blast resistance and identification of resistance genes in main indica rice patent materials in South China,Nanfang Nongye Xuebao(Journal of Southern Agriculture),48(4):587-593(阎勇,马增凤,秦钢,陈远孟,秦媛媛,颜群,刘驰,张月雄,黄大辉,2017,华南常用籼稻亲本稻瘟病抗性评价及抗性基因鉴定,南方农业学报,48(4):587-593)
Zhang Y.,Feng Z.F.,Cui M.Z.,and Zhang X.J.,2013a,Distribution of blast resistant gene Pi-km in rice germplasm in Shanxi province of northwest China,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),11(3):311-316(张羽,冯志峰,崔明珠,张晓娟,2013a,稻瘟病抗性基因Pi-km在陕西省稻种资源中的分布状况,分子植物育种,11(3):311-316)
Zhang Y.,Feng Z.F.,Zhang H.,and Mo D.,2013b,Distribution of blast resistant gene Pi9 in rice germplasm in Shanxi Province,Sichuan Nongye Daxue Xuebao(Journal of Sichuan Agricultural University),31(2):115-121(张羽,冯志峰,张晗,莫丹,2013b,陕西省水稻种质资源中Pi9基因的分布状况,四川农业大学学报,31(2):115-121)
Zou D.T.,Jiang S.D.,Zhao H.W.,Guo L.Y.,Sun J.,Liu H.L.,and Xin W.,2016,Distribution of broad-spectrum resistance gene-Pi9 in the rice cultivars of Heilongjiang Province,Dongbei Nongye Daxue Xuebao(Journal of Northeast Agricultural University),47(7):1-8(邹德堂,姜思达,赵宏伟,郭丽颖,孙健,刘化龙,辛威,2016,广谱抗性基因Pi9在黑龙江省水稻品种中的分布,东北农业大学学报,47(7):1-8)