水稻细胞质雄性不育系谷丰A稻瘟病抗性基因分子鉴定及遗传分析
|
摘要
谷丰A是一个优良的抗稻瘟病水稻细胞质雄性不育系,利用它已配制出多个优良杂交水稻品种,并且已成为选育抗稻瘟病不育系的重要亲本之一。本研究利用Pigm、Pi9、Pi2、Piz-t、Pi-d2、Pi-d3、Pit、Pi5、Pii、Pib、Pi-ta等11个抗稻瘟病基因的特异分子标记,检测和分析了谷丰A的抗稻瘟病基因型。结果表明,该不育系中含有稻瘟病抗性基因Pi-d2、Pi-d3和Pigm。进一步分析确定了这3个基因都来源于其亲本之一地谷A。用8个稻瘟病菌田间接种鉴定表明,谷丰A比福伊B及单基因系谷梅4号、IRBLKs-Ka (Pik)、IRBL9-W (Pi9)、IRBLta-K1 (Pi-ta)等广谱高抗的材料具有更好的抗性。
GuFeng A is a good rice cytoplasmic male sterile line with resistance to rice blast, many hybrid rice varieties have been bred by using GuFeng A, and it has become one of the most important parents for breeding the rice blast-resistant sterile line. In this paper, the specific molecular primers of eleven rice blast resistant genes, such as Pigm, Pi9, Pi2, Piz-t, Pi-d2, Pi-d3, Pit, Pi5, Pii, Pib, Pi-ta, were used to detect and analyze the rice blast-resistant genotype of Gu Feng A. The results indicated that the sterile line contained three rice blast resistance genes Pi-d2,Pi-d3 and Pigm. The further analysis confirmed that all of these three genes were derived from one of their parents DiGu A. Inoculation identification of eight Magnaporthe oryzae in the field confirmed that the resistance to blast of Gu Feng A is better than the broad spectrum and highly resistant materials FuYi B and single gene lines GuMei 4 Hao, IRBLKs-Ka(Pik), IRBL9-W(Pi9) and IRBLta-K1(Pi-ta).
GuFeng A is a good rice cytoplasmic male sterile line with resistance to rice blast, many hybrid rice varieties have been bred by using GuFeng A, and it has become one of the most important parents for breeding the rice blast-resistant sterile line. In this paper, the specific molecular primers of eleven rice blast resistant genes, such as Pigm, Pi9, Pi2, Piz-t, Pi-d2, Pi-d3, Pit, Pi5, Pii, Pib, Pi-ta, were used to detect and analyze the rice blast-resistant genotype of Gu Feng A. The results indicated that the sterile line contained three rice blast resistance genes Pi-d2,Pi-d3 and Pigm. The further analysis confirmed that all of these three genes were derived from one of their parents DiGu A. Inoculation identification of eight Magnaporthe oryzae in the field confirmed that the resistance to blast of Gu Feng A is better than the broad spectrum and highly resistant materials FuYi B and single gene lines GuMei 4 Hao, IRBLKs-Ka(Pik), IRBL9-W(Pi9) and IRBLta-K1(Pi-ta).
引文
Bryan G.T.,Wu K.S.,Farrall L.,Jia Y.L.,Hershey H.P.,McAdams S.A.,Faulk K.N.,Donaldson G.K.,Tarchini R.,and Valent B.,2000,A single amino acid difference distinguishes resistant and susceptible alleles of the rice blast resistance gene Pi-ta,Plant Cell,12(11):2033-2045
Chen X.W.,Shang J.J.,Chen D.X.,Lei C.L.,Zou Y.,Zhai W.X.,Liu G.Z.,Xu J.C.,Ling Z.Z.,Cao G.,Ma B.T.,Wang Y.P.,Zhao X.F.,Li S.G.,and Zhu L.H.,2006,A B-lectin receptor kinase gene conferring rice blast resistance,Plant J.,46(5):794-804
Chen X.W.,Li S.G.,Xu J.C.,Zhai W.X.,Ling Z.Z.,Ma B.T.,Wang Y.P.,Wang W.M.,Cao G.,Ma Y.Q.,Shang J.J.,Zhao X.F.,Zhou K.D.,and Zhu L.H.,2004,Identification of two blast resistance genes in a rice variety,Digu,J.Phytopathol.,152(2):77-85
Delseny M.,Salses J.,Cooke R.,Sallaud C.,Regad F.,Lagoda P.,Guiderdoni E.,Ventelon M.,Brugidou C.,and Ghesquière A.,2001,Rice genomics:present and future,Plant Physiol.Biochem.,39(3-4):323-334
Deng Y.W.,Zhai K.R.,Xie Z.,Yang D.Y.,Zhu X.D.,Liu J.Z.,Wang X.,Qin P.,Yang Y.Z.,Zhang G.M.,Li Q.,Zhang J.F.,Wu S.Q.,Milazzo J.,Mao B.Z.,Wang E.,Xie H.A.,Tharreau D.,and He Z.H.,2017,Epigenetic regulation of antagonistic receptors confers rice blast resistance with yield balance,Science,355(6328):962-965
Gao L.J.,Deng G.F.,Gao H.L.,Gao G.Q.,Zhou M.,Zhou W.Y.,Yan Q.,Zhang J.,and Dai G.X.,2010,Establishment and application of gene tagging linked to rice blast resistance gene pi-d2,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),23(1):77-82(高利军,邓国富,高汉亮,高国庆,周萌,周维永,颜群,张晋,戴高兴,2010,水稻抗稻瘟病基因pi-d2基因标签的建立与应用,西南农业学报,23(1):77-82)
Hayashi K.,Yasuda N.,Fujita Y.,Koizumi S.,and Yoshida H.,2010,Identification of the blast resistance gene pit in rice cultivars using functional markers,Theor.Appl.Genet.,121(7):1357-1367
Huang L.X.,Liang K.J.,You N.S.,Wu Z.Y.,and Huang S.,2011,Classical genetic analysis of serial rice parents with rice blast resistance genes,Fujian Nonglin Daxue Xuebao(Journal of Fujian Agriculture and Forestry University(Natural Science Edition)),40(3):225-230(黄利兴,梁康迳,游年顺,吴志源,黄姗,2011,系列抗稻瘟病水稻亲本抗性基因的经典遗传学分析,福建农林大学学报(自然科学版),40(3):225-230)
Huang L.X.,Zhang Y.H.,You N.S.,Hu R.H.,Wu Z.Y.,Lei S.P.,Lei J.C.,Pan Y.Q.,and Xie M.Z.,2013,Breeding and utilization of rice fertility line Gufeng A with blast resistance,Zajiao Shuidao(Hybrid Rice),28(5):6-10(黄利兴,张以华,游年顺,胡荣华,吴志源,雷上平,雷捷成,潘玉卿,谢美珠,2013,抗稻瘟病水稻不育系谷丰A的选育与利用,杂交水稻,28(5):6-10)
Lee S.K.,Song M.Y.,Seo Y.S.,Kim H.K.,Ko S.,Cao P.J.,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 coiled coil nucleotide binding-leucine rich repeat genes,Genetics,181(4):1627-1638
Liu Y.,Xu P.Z.,Zhang H.Y.,Xu J.D.,Wu F.Q.,and Wu X.J.,2008,Marker-assisted selection and application of blast resistant gene Pib in rice,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),41(1):9-14(刘洋,徐培洲,张红宇,徐建第,吴发强,吴先军,2008,水稻抗稻瘟病Pib基因的分子标记辅助选择与应用,中国农业科学,41(1):9-14)
Liu J.L.,Wang X.J.,Mitchell T.,Hu Y.J.,Liu X.L.,Dai L.Y.,and Wang G.L.,2010,Recent progress and understanding of the molecular mechanisms of the rice-Magnaporthe oryzae interaction,Mol.Plant Pathol.,11(3):419-427
Miah G.,Rafii M.Y.,Ismail M.R.,Puteh A.B.,Rahim H.A.,Asfaliza R.,and Latif M.A.,2013,Blast resistance in rice:a review of conventional breeding to molecular approaches,Mol.Biol.Rep.,40(3):2369-2388
Shang J.J.,Tao Y.,Chen X.W.,Zou Y.,Lei C.L.,Wang J.,Li X.B.,Zhao X.F.,Zhang M.J.,Lu Z.K.,Xu J.C.,Cheng Z.K.,Wan J.M.,and Zhu L.H.,2009,Identification of a new rice blast resistance gene,Pid3,by genomewide comparison of paired nucleotide-binding site-leucine-rich repeat genes and their pseudogene alleles between the two sequenced rice genomes,Genetics,182(4):1303-1311
Tian D.G.,Chen Z.J.,Chen Z.Q.,Zhou Y.C.,Wang Z.H.,Wang F.,and Chen S.B.,2016,Allele-specific marker-based assessment revealed that the rice blast resistance genes Pi2 and Pi9 have not been widely deployed in Chinese indica rice cultivars,Rice,9(1):19
Takagi H.,Uemura A.,Yaegashi H.,Tamiru M.,Abe A.,Mitsuoka C.,Utsushi H.,Natsume S.,Kanzaki H.,Matsumura H.,Saitoh H.,Yoshida K.,Cano L.M.,Kamoun S.O.,and Terauchi R.,2013,MutMap-Gap:whole-genome resequencing of mutant F2progeny bulk combined with de novo assembly of gap regions identifies the rice blast resistance gene Pii,New Phytol.,200(1):276-283
Wang Z.X.,Yano M.,Yamanouchi U.,Iwamoto M.,Monna L.,Hayasaka H.,Katayose Y.,and Sasaki T.,1999,The Pib gene for rice blast resistance belongs to the nucleotide binding and leucine-rich repeat class of plant disease resistance genes,Plant J.,19(1):55-64
Yi G.,Lee S.K.,Hong Y.K.,Cho Y.C.,Nam M.H.,Kim S.C.,Han G.L.,Hahn T.R.,Ronald P.C.,and Jeon J.S.,2004,Use of Pi5(t)markers in marker-assisted selection to screen for cultivars with resistance to Magnaporthe grisea,Theor.Appl.Genet.,109(5):978-985
Zeng S.Y.,Gong H.B.,Li C.,Du C.C.,Sun L.T.,Jin D.D.,Lin T.Z.,Yu B.,Qian H.F.,Yao W.C.,and Zhou Y.W.,2016,Molecular markers of rice blast resistance gene Pigm and its application in GuMei 4,China Patent,CN106148335A(曾生元,龚红兵,李闯,杜灿灿,孙立亭,景德道,林添资,余波,钱华飞,姚维成,周义文,2016,谷梅4号抗稻瘟病基因Pigm的分子标记及其应用,中国专利,CN106148335A)
Zhang F.J.,Zhang M.L.,and Zhu S.F.,2004,An improved rapid method of plant total DNA extraction,Zhiwu Jianyi(Plant Quarantine),18(6):330-332(张凤娟,张满良,朱水芳,2004,一种改进的水稻总DNA的快速提取方法,植物检疫,18(6):330-332)
Zhou M.J.,Yi C.C.,and Zheng A.P.,2016,PCR-based molecular marker detection of the gene resistant to rice blast in sterile line'GangXiang 1A',Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(4):941-945(周明镜,尹传春,郑爱萍,2016,不育系“冈香1A”抗稻瘟病基因的分子检测,分子植物育种,14(4):941-945)
Zhang Y.,Zhang X.J.,Yang F.J.,and Feng Z.F.,2013,SNP-based detecting method of rice blast resistance gene Pi-ta,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),27(3):325-328(张羽,张晓娟,杨凤娇,冯志峰,2013,水稻稻瘟病抗性基因Pi-ta的SNP检测方法,中国水稻科学,27(3):325-328)
Chen X.W.,Shang J.J.,Chen D.X.,Lei C.L.,Zou Y.,Zhai W.X.,Liu G.Z.,Xu J.C.,Ling Z.Z.,Cao G.,Ma B.T.,Wang Y.P.,Zhao X.F.,Li S.G.,and Zhu L.H.,2006,A B-lectin receptor kinase gene conferring rice blast resistance,Plant J.,46(5):794-804
Chen X.W.,Li S.G.,Xu J.C.,Zhai W.X.,Ling Z.Z.,Ma B.T.,Wang Y.P.,Wang W.M.,Cao G.,Ma Y.Q.,Shang J.J.,Zhao X.F.,Zhou K.D.,and Zhu L.H.,2004,Identification of two blast resistance genes in a rice variety,Digu,J.Phytopathol.,152(2):77-85
Delseny M.,Salses J.,Cooke R.,Sallaud C.,Regad F.,Lagoda P.,Guiderdoni E.,Ventelon M.,Brugidou C.,and Ghesquière A.,2001,Rice genomics:present and future,Plant Physiol.Biochem.,39(3-4):323-334
Deng Y.W.,Zhai K.R.,Xie Z.,Yang D.Y.,Zhu X.D.,Liu J.Z.,Wang X.,Qin P.,Yang Y.Z.,Zhang G.M.,Li Q.,Zhang J.F.,Wu S.Q.,Milazzo J.,Mao B.Z.,Wang E.,Xie H.A.,Tharreau D.,and He Z.H.,2017,Epigenetic regulation of antagonistic receptors confers rice blast resistance with yield balance,Science,355(6328):962-965
Gao L.J.,Deng G.F.,Gao H.L.,Gao G.Q.,Zhou M.,Zhou W.Y.,Yan Q.,Zhang J.,and Dai G.X.,2010,Establishment and application of gene tagging linked to rice blast resistance gene pi-d2,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),23(1):77-82(高利军,邓国富,高汉亮,高国庆,周萌,周维永,颜群,张晋,戴高兴,2010,水稻抗稻瘟病基因pi-d2基因标签的建立与应用,西南农业学报,23(1):77-82)
Hayashi K.,Yasuda N.,Fujita Y.,Koizumi S.,and Yoshida H.,2010,Identification of the blast resistance gene pit in rice cultivars using functional markers,Theor.Appl.Genet.,121(7):1357-1367
Huang L.X.,Liang K.J.,You N.S.,Wu Z.Y.,and Huang S.,2011,Classical genetic analysis of serial rice parents with rice blast resistance genes,Fujian Nonglin Daxue Xuebao(Journal of Fujian Agriculture and Forestry University(Natural Science Edition)),40(3):225-230(黄利兴,梁康迳,游年顺,吴志源,黄姗,2011,系列抗稻瘟病水稻亲本抗性基因的经典遗传学分析,福建农林大学学报(自然科学版),40(3):225-230)
Huang L.X.,Zhang Y.H.,You N.S.,Hu R.H.,Wu Z.Y.,Lei S.P.,Lei J.C.,Pan Y.Q.,and Xie M.Z.,2013,Breeding and utilization of rice fertility line Gufeng A with blast resistance,Zajiao Shuidao(Hybrid Rice),28(5):6-10(黄利兴,张以华,游年顺,胡荣华,吴志源,雷上平,雷捷成,潘玉卿,谢美珠,2013,抗稻瘟病水稻不育系谷丰A的选育与利用,杂交水稻,28(5):6-10)
Lee S.K.,Song M.Y.,Seo Y.S.,Kim H.K.,Ko S.,Cao P.J.,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 coiled coil nucleotide binding-leucine rich repeat genes,Genetics,181(4):1627-1638
Liu Y.,Xu P.Z.,Zhang H.Y.,Xu J.D.,Wu F.Q.,and Wu X.J.,2008,Marker-assisted selection and application of blast resistant gene Pib in rice,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),41(1):9-14(刘洋,徐培洲,张红宇,徐建第,吴发强,吴先军,2008,水稻抗稻瘟病Pib基因的分子标记辅助选择与应用,中国农业科学,41(1):9-14)
Liu J.L.,Wang X.J.,Mitchell T.,Hu Y.J.,Liu X.L.,Dai L.Y.,and Wang G.L.,2010,Recent progress and understanding of the molecular mechanisms of the rice-Magnaporthe oryzae interaction,Mol.Plant Pathol.,11(3):419-427
Miah G.,Rafii M.Y.,Ismail M.R.,Puteh A.B.,Rahim H.A.,Asfaliza R.,and Latif M.A.,2013,Blast resistance in rice:a review of conventional breeding to molecular approaches,Mol.Biol.Rep.,40(3):2369-2388
Shang J.J.,Tao Y.,Chen X.W.,Zou Y.,Lei C.L.,Wang J.,Li X.B.,Zhao X.F.,Zhang M.J.,Lu Z.K.,Xu J.C.,Cheng Z.K.,Wan J.M.,and Zhu L.H.,2009,Identification of a new rice blast resistance gene,Pid3,by genomewide comparison of paired nucleotide-binding site-leucine-rich repeat genes and their pseudogene alleles between the two sequenced rice genomes,Genetics,182(4):1303-1311
Tian D.G.,Chen Z.J.,Chen Z.Q.,Zhou Y.C.,Wang Z.H.,Wang F.,and Chen S.B.,2016,Allele-specific marker-based assessment revealed that the rice blast resistance genes Pi2 and Pi9 have not been widely deployed in Chinese indica rice cultivars,Rice,9(1):19
Takagi H.,Uemura A.,Yaegashi H.,Tamiru M.,Abe A.,Mitsuoka C.,Utsushi H.,Natsume S.,Kanzaki H.,Matsumura H.,Saitoh H.,Yoshida K.,Cano L.M.,Kamoun S.O.,and Terauchi R.,2013,MutMap-Gap:whole-genome resequencing of mutant F2progeny bulk combined with de novo assembly of gap regions identifies the rice blast resistance gene Pii,New Phytol.,200(1):276-283
Wang Z.X.,Yano M.,Yamanouchi U.,Iwamoto M.,Monna L.,Hayasaka H.,Katayose Y.,and Sasaki T.,1999,The Pib gene for rice blast resistance belongs to the nucleotide binding and leucine-rich repeat class of plant disease resistance genes,Plant J.,19(1):55-64
Yi G.,Lee S.K.,Hong Y.K.,Cho Y.C.,Nam M.H.,Kim S.C.,Han G.L.,Hahn T.R.,Ronald P.C.,and Jeon J.S.,2004,Use of Pi5(t)markers in marker-assisted selection to screen for cultivars with resistance to Magnaporthe grisea,Theor.Appl.Genet.,109(5):978-985
Zeng S.Y.,Gong H.B.,Li C.,Du C.C.,Sun L.T.,Jin D.D.,Lin T.Z.,Yu B.,Qian H.F.,Yao W.C.,and Zhou Y.W.,2016,Molecular markers of rice blast resistance gene Pigm and its application in GuMei 4,China Patent,CN106148335A(曾生元,龚红兵,李闯,杜灿灿,孙立亭,景德道,林添资,余波,钱华飞,姚维成,周义文,2016,谷梅4号抗稻瘟病基因Pigm的分子标记及其应用,中国专利,CN106148335A)
Zhang F.J.,Zhang M.L.,and Zhu S.F.,2004,An improved rapid method of plant total DNA extraction,Zhiwu Jianyi(Plant Quarantine),18(6):330-332(张凤娟,张满良,朱水芳,2004,一种改进的水稻总DNA的快速提取方法,植物检疫,18(6):330-332)
Zhou M.J.,Yi C.C.,and Zheng A.P.,2016,PCR-based molecular marker detection of the gene resistant to rice blast in sterile line'GangXiang 1A',Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(4):941-945(周明镜,尹传春,郑爱萍,2016,不育系“冈香1A”抗稻瘟病基因的分子检测,分子植物育种,14(4):941-945)
Zhang Y.,Zhang X.J.,Yang F.J.,and Feng Z.F.,2013,SNP-based detecting method of rice blast resistance gene Pi-ta,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),27(3):325-328(张羽,张晓娟,杨凤娇,冯志峰,2013,水稻稻瘟病抗性基因Pi-ta的SNP检测方法,中国水稻科学,27(3):325-328)