雪茄烟品种Beinhart1000-1赤星病抗性基因的QTL定位
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摘要
由链格孢菌(Alternariaalternata)引起的赤星病是最具破坏性的烟草叶斑病害之一,在中国严重地影响烟草(Nicotiana tabacum L.)的产量和质量。选育抗烟草赤星病的优良品种虽是预防该病最经济、有效的途径,但因其抗性受数量性状基因控制而很难通过常规育种手段实现。为了便于开展烟草抗赤星病分子标记辅助选择育种,本研究利用抗赤星病雪茄烟品种Beinhart1000-1和感病烤烟品种红花大金元经杂交、自交产生的362个F_2单株构建了一张包含670个SSR标记的烟草遗传连锁图谱,并结合组培快繁形成的F_2株系的田间赤星病病情指数(DI),在全基因组范围内检测获得2个与烟草赤星病抗性相关的QTL,分别位于第20和23连锁群上的SSR标记TMs05179和TMs04022,以及TM61049和TM62212之间。这2个QTL等位基因均来自抗病亲本Beinhart1000-1,它们一起解释了两亲本间81%的病情指数(DI)差异及64%的加性效应。为下一步开展烟草抗赤星病的分子标记辅助育种奠定了基础。
Tobacco brown spot(TBS) caused by Alternaria alternata is one of the most destructive foliar diseases affecting tobacco(Nicotiana tabacum L.) production and quality in China. Breeding TBS-resistant cultivars is difficult by traditional method because the resistance has proved to be quantitatively inherited. To facilitate marker-assisted selection, we carried out a study of mapping quantitative trait loci(QTLs) for TBS resistance. We developed an F_2 population consisting of 362 individuals from a cross between a TBS-susceptive flue-cured tobacco Honghua Dajinyuan(HD) and a TBS-resistant cigar tobacco cultivar Beinhart1000-1, and constructed a genetic map consisting of 670 SSR markers based on this population. Using disease index(DI) as the indicator of TBS resistance, we detected two QTLs located between SSR markers TMs05179 and TMs04022, and TM61049 and TM62212 on linkage group(LG) 20 and LG23, respectively. The resistant alleles of the two QTLs were all from the resistant parent Beinhart1000-1. The two QTLs together could explain 81% of the DI difference between the two parents in total, and 64%of their additive effects. Therefore, the two QTLs will be useful for TBS resistance breeding.
Tobacco brown spot(TBS) caused by Alternaria alternata is one of the most destructive foliar diseases affecting tobacco(Nicotiana tabacum L.) production and quality in China. Breeding TBS-resistant cultivars is difficult by traditional method because the resistance has proved to be quantitatively inherited. To facilitate marker-assisted selection, we carried out a study of mapping quantitative trait loci(QTLs) for TBS resistance. We developed an F_2 population consisting of 362 individuals from a cross between a TBS-susceptive flue-cured tobacco Honghua Dajinyuan(HD) and a TBS-resistant cigar tobacco cultivar Beinhart1000-1, and constructed a genetic map consisting of 670 SSR markers based on this population. Using disease index(DI) as the indicator of TBS resistance, we detected two QTLs located between SSR markers TMs05179 and TMs04022, and TM61049 and TM62212 on linkage group(LG) 20 and LG23, respectively. The resistant alleles of the two QTLs were all from the resistant parent Beinhart1000-1. The two QTLs together could explain 81% of the DI difference between the two parents in total, and 64%of their additive effects. Therefore, the two QTLs will be useful for TBS resistance breeding.
引文
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[2]Main C E.The tobacco brown spot lesion,a model to study halo formation.Phytopathology,1969,59:1040-1045.
[3]Deborah R,Harvey W.Biocontrol of tobacco brown-spot disease by bacillus cereus in a controlled environment.Phytopathology,1991,61:930-932.
[4]Ramm C V,Lucus G B.Epiphytology of tobacco brown spot caused by Alternaria longipes.Phytopathology,1963,53:450-455.
[5]Stavely J R,Slana L J.Relation of leaf age to the reaction of tobacco to Alternaria alternate.Phytopathology,1970,61:73-78.
[6]Slavov S,Mayama S,Atamassov A.Toxin production of Alternaria alternata tobacco pathotype.Biotechnol Biotechnol Equip,2004,18:90-95.
[7]Jenning D B,Daub M E,Pharr D M,Williamson J D.Constitutive expression of a celery mannitol dehydrogenase in tobacco enhances resistance to the mannitol-secreting fungal pathogen Alternaria alternata.Plant J,2002,32:41-49.
[8]Yakinova E T,Yordanova Z P,Slavov S,Toteva V M K,Woltering E J.Alternaria alternata AT toxin induces programmed cell death in tobacco.J Phytopathol,2009,157:592-601.
[9]Stavely J R.Inheritance of brown spot resistance in Nicotiana tabacum.Proc Am Phytopathol Soc,1975,2:228-230.
[10]Dobhal V K,Monga D.Genetic analysis of field resistance to brown spot caused by Alternaria altemata(Fries)Keisaler in Nicotiana rustica L.Tobacco Res,1991,17:11-15.
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[13]Stuber C W,Edwards M D,Wendel J F.Molecular markerfacilitated investigations of quantitative trait loci in maize:II.Factors influencing yield and its component traits.Crop Sci,1987,27:639-648.
[14]Nishi T,Tajima T,Noguchi S,Ajisaka H,Negishi H.Identification of DNA markers of tobacco linked to bacterial wilt resistance.Theor Appl Genet,2003,106:765-770.
[15]Julio E,Denoyes R B,Verrier J L,de Borne F D.Detection of QTLs linked to leaf and smoke properties in Nicotiana tabacum based on a study of 114 recombinant inbred lines.Mol Breed,2006,22:144-166.
[16]Vontimitta V,Lewis R S.Mapping of quantitative trait loci affecting resistance to Phytophthora nicotianae in tobacco(Nicotiana tabacum L.)line Beihart-1000.Mol Breed,2010,58:294-300.
[17]Tong Z J,Jiao T L,Wang F Q,Li M Y,Leng X D,Gao Y L,Li YP,Xiao B G,Wu W R.Mapping of quantitative trait loci conferring resistance to brown spot in flue-cured tobacco(Nicotiana tabacum L.).Plant Breed,2012,131:335-339.
[18]蒋彩虹,罗成刚,任民,杨爱国,冯全福,王元英.一个与净叶黄抗赤星病基因紧密连锁的SSR标记.中国烟草科学,2012,33(1):9-22.Jiang C H,Luo C G,Ren M,Yang A G,Feng Q F,Wang Y Y.ASSR marker tightly linked to the resistant gene of Jingyehuang on tobacco brown spot.Chin Tobacco Sci,2012,33(1):19-22(in Chinese with English abstract).
[19]蒋彩虹,王元英,任民,张兴伟,杨爱国,程立锐,冯全福,罗成刚.一个抗赤星病基因的SSR标记连锁群.分子植物育种,2013,11:566-569.Jiang C H,Wang Y Y,Ren M,Zhang X W,Yang A G,Cheng L R,Feng Q F,Luo C G.A linkage group of SSR markers linked to the resistant gene on tobacco brown spot.Mol Plant Breed,2013,11:566-569(in Chinese with English abstract).
[20]高亭亭,蒋彩虹,罗成刚,杨爱国,程立锐,代帅帅.Beinhart1000-1抗赤星病基因的QTL定位.中国烟草学报,2014,20(2):104-107.Gao T T,Jiang C H,Luo C G,Yang A G,Cheng L R,Dai S S.Mapping of quantitative trait loci affecting resistance to brown spot in tobacco line Beinhart1000-1.Acta Tobacco Sin,2014,20(2):104-107(in Chinese with English abstract).
[21]Del Piano L,Abet M,Sorrentino C,Acanfora F,Cozzolino E,DiMuro A.Genetic variability in Nicotiana tabacum and Nicotiana species as revealed by RAPD procedure.Int Contribu Tobacco Res,2000,19:1-15.
[22]Ren N,Timko M P.ALFP analysis of genetic polymorphism and evolutionary relationships among cultivated and wild Nicotiana species.Genome,2001,44:559-571.
[23]Rossi L,Bindler G,Pijnenburg H,Isaac P G,Henri I G,Mahe M,Orvain C,Gadani F.Potential of molecular marker analysis for variety identification in processed tobacco.Plant Varieties Seeds,2001,14:89-101.
[24]Arslan B,Okunus A.Genetic and geographic polymorphism of cultivated tobaccos(Nicotiana tabacum)in Turkey.Plant Genetics,2006,42:667-671.
[25]Julio E,Verrier J L,de Borne F D.Development of SCAR markers linked to three disease resistances based on AFLP within Nicotiana tabacum L.Theor Appl Genet,2006,112:335-346.
[26]Moon H S,Nicholson J S,Lewis R S.Use of transferable Nicotiana tabacum L.microsatellite markers for investigating genetic diversity in the genus Nicotiana.Genome,2008,51:547-559.
[27]Raju K S,Madhav M S,Sharma R K,Murthy T G K,Mohapatra T.Genetic polymorphism of Indian tobacco types as revealed by amplified fragment length polymorphism.Curr Sci,2008,94:633-638.
[28]Moon H S,Nicholson J S,Heineman A,Lion K,der Hoeven R V,Hayes A J,Lewis R S.Changes in genetic diversity of U.S.flue-cured tobacco germplasm over seven decades of cultivar development.Crop Sci,2009,49:498-506.
[29]Moon H S,Nifong J M,Nicholson J S,Heineman A,Lion K,der Hoeven R V,Hayes A J,Lewis R S.Microsatellite-based analysis of tobacco(Nicotiana tabacum L.)genetic resources.Crop Sci,2009,49:2149-2157.
[30]Bindler G,der Hoeven V R,Gunduz I,Plieske J,Ganal M,Rossi L,Gadani F,Donini P.A microsatellite marker based linkage map of tobacco.Theor Appl Genet,2007,114:341-349.
[31]Bindler G,Plieske J,Bakaher N,Gunduz I,Ivanov N,der Hoeven R V,Ganal M,Donini P.A high density genetic map of tobacco(Nicotiana tabacum L.)obtained from large scale microsatellite marker development.Theor Appl Genet,2011,123:219-230.
[32]Tong Z J,Yang Z M,Chen X J,Jiao F C,Li X Y,Wu X F,Gao YL,Xiao B G,Wu W R.Large-scale development of microsatellite markers in Nicotiana tabacum and construction of a genetic map of flue-cured tobacco.Plant Breed,2012,131:674-680.
[33]Tong Z J,Xiao B G,Jiao F C,Fang D H,Zeng J M,Wu X F,Chen X J,Yang J K,Li Y P.Large-scale development of SSRmarkers in tobacco and construction of a linkage map in flue-cured tobacco.Breed Sci,2016,66:381-390.
[34]郭永峰,朱贤朝,孔凡玉,石金开,王年.赤星病抗源的抗性比较.中国烟草科学,1997,11(4):1-6.Guo Y F,Zhu X C,Kong F Y,Shi J K,Wang N.Comparing two origins of resistance to brown spot disease in tobacco.Chin Tobacco Sci,1997,11(4):1-6(in Chinese with English abstract).
[35]许绍斌,陶玉芬,杨昭庆,褚嘉.简单快速的DNA银染和胶保存方法.遗传,2002,24:335-336.Xu S B,Tao Y F,Yang Z Q,Chu J.A simple and rapid methods used for silver staining and gel preservation.Hereditas,2002,24:335-336(in Chinese with English abstract).
[36]Van Ooijen J W.JoinMap?4.0,Software for the calculation of genetic linkage maps in experimental populations.Wageningen:Kyazma B.V Press,2006.pp 1-63.
[37]Voorrips R E.MapChart:Software for the graphical presentation of linkage maps and QTLs.J Heredity,2002,93:77-78.
[38]Van Ooijen J W.MapQTL?6.0,Software for the mapping of quantitative trait loci in experimental populations of diploid species.Wageningen:Kyazma B V Press,2009.pp 1-64.
[39]Mc Couch S R,Cho Y G,Yano M,Paul E,Blinstrub M,Morishima H,Kinoshita T.Report on QTL nomenclature.Rice Genet Newsl,1997,14:11-13.
[40]冯莹,蒋彩虹,程立锐,杨爱国,郑吉云,赵清海,杨修峰,尹华玲,冯全福.两个烟草赤星病抗源的遗传分析.中国烟草科学,2015,36(5):1-7.Feng Y,Jiang C H,Cheng L R,Yang A G,Zheng J Y,Zhao QH,Yang X F,Yin H L,Feng Q F.Genetic analysis of resistance to brown spot disease in tobacco cultivars Jingyehuang and Beinhart1000-1.Chin Tobacco Sci,2015,36(5):1-7(in Chinese with English abstract).
[2]Main C E.The tobacco brown spot lesion,a model to study halo formation.Phytopathology,1969,59:1040-1045.
[3]Deborah R,Harvey W.Biocontrol of tobacco brown-spot disease by bacillus cereus in a controlled environment.Phytopathology,1991,61:930-932.
[4]Ramm C V,Lucus G B.Epiphytology of tobacco brown spot caused by Alternaria longipes.Phytopathology,1963,53:450-455.
[5]Stavely J R,Slana L J.Relation of leaf age to the reaction of tobacco to Alternaria alternate.Phytopathology,1970,61:73-78.
[6]Slavov S,Mayama S,Atamassov A.Toxin production of Alternaria alternata tobacco pathotype.Biotechnol Biotechnol Equip,2004,18:90-95.
[7]Jenning D B,Daub M E,Pharr D M,Williamson J D.Constitutive expression of a celery mannitol dehydrogenase in tobacco enhances resistance to the mannitol-secreting fungal pathogen Alternaria alternata.Plant J,2002,32:41-49.
[8]Yakinova E T,Yordanova Z P,Slavov S,Toteva V M K,Woltering E J.Alternaria alternata AT toxin induces programmed cell death in tobacco.J Phytopathol,2009,157:592-601.
[9]Stavely J R.Inheritance of brown spot resistance in Nicotiana tabacum.Proc Am Phytopathol Soc,1975,2:228-230.
[10]Dobhal V K,Monga D.Genetic analysis of field resistance to brown spot caused by Alternaria altemata(Fries)Keisaler in Nicotiana rustica L.Tobacco Res,1991,17:11-15.
[11]郭永峰,朱贤朝,石金开,孔凡玉,王年,王从丽.烟草对赤星病田间抗性的遗传研究.中国烟草科学,1998,17(3):11-16.Guo Y F,Zhu X C,Shi J K,Kong F Y,Wang N,Wang C L.Genetic studies on resistance to brown spot disease in tobacco.Chin Tobacco Sci,17(3):11-16(in Chinese with English abstract).
[12]郭永峰,石金开,孔凡玉,王年,王从丽,何京美,朱贤朝.烟草赤星病抗性因素遗传的双列分析.中国烟草科学,2000,21(4):17-20.Guo Y F,Shi J K,Kong F Y,Wang N,He J M,Zhu X C.Diallel cross analysis on inheritance of resistance components to tobacco brown spot disease.Chin Tobacco Sci,2000,21(4):17-20(in Chinese with English abstract).
[13]Stuber C W,Edwards M D,Wendel J F.Molecular markerfacilitated investigations of quantitative trait loci in maize:II.Factors influencing yield and its component traits.Crop Sci,1987,27:639-648.
[14]Nishi T,Tajima T,Noguchi S,Ajisaka H,Negishi H.Identification of DNA markers of tobacco linked to bacterial wilt resistance.Theor Appl Genet,2003,106:765-770.
[15]Julio E,Denoyes R B,Verrier J L,de Borne F D.Detection of QTLs linked to leaf and smoke properties in Nicotiana tabacum based on a study of 114 recombinant inbred lines.Mol Breed,2006,22:144-166.
[16]Vontimitta V,Lewis R S.Mapping of quantitative trait loci affecting resistance to Phytophthora nicotianae in tobacco(Nicotiana tabacum L.)line Beihart-1000.Mol Breed,2010,58:294-300.
[17]Tong Z J,Jiao T L,Wang F Q,Li M Y,Leng X D,Gao Y L,Li YP,Xiao B G,Wu W R.Mapping of quantitative trait loci conferring resistance to brown spot in flue-cured tobacco(Nicotiana tabacum L.).Plant Breed,2012,131:335-339.
[18]蒋彩虹,罗成刚,任民,杨爱国,冯全福,王元英.一个与净叶黄抗赤星病基因紧密连锁的SSR标记.中国烟草科学,2012,33(1):9-22.Jiang C H,Luo C G,Ren M,Yang A G,Feng Q F,Wang Y Y.ASSR marker tightly linked to the resistant gene of Jingyehuang on tobacco brown spot.Chin Tobacco Sci,2012,33(1):19-22(in Chinese with English abstract).
[19]蒋彩虹,王元英,任民,张兴伟,杨爱国,程立锐,冯全福,罗成刚.一个抗赤星病基因的SSR标记连锁群.分子植物育种,2013,11:566-569.Jiang C H,Wang Y Y,Ren M,Zhang X W,Yang A G,Cheng L R,Feng Q F,Luo C G.A linkage group of SSR markers linked to the resistant gene on tobacco brown spot.Mol Plant Breed,2013,11:566-569(in Chinese with English abstract).
[20]高亭亭,蒋彩虹,罗成刚,杨爱国,程立锐,代帅帅.Beinhart1000-1抗赤星病基因的QTL定位.中国烟草学报,2014,20(2):104-107.Gao T T,Jiang C H,Luo C G,Yang A G,Cheng L R,Dai S S.Mapping of quantitative trait loci affecting resistance to brown spot in tobacco line Beinhart1000-1.Acta Tobacco Sin,2014,20(2):104-107(in Chinese with English abstract).
[21]Del Piano L,Abet M,Sorrentino C,Acanfora F,Cozzolino E,DiMuro A.Genetic variability in Nicotiana tabacum and Nicotiana species as revealed by RAPD procedure.Int Contribu Tobacco Res,2000,19:1-15.
[22]Ren N,Timko M P.ALFP analysis of genetic polymorphism and evolutionary relationships among cultivated and wild Nicotiana species.Genome,2001,44:559-571.
[23]Rossi L,Bindler G,Pijnenburg H,Isaac P G,Henri I G,Mahe M,Orvain C,Gadani F.Potential of molecular marker analysis for variety identification in processed tobacco.Plant Varieties Seeds,2001,14:89-101.
[24]Arslan B,Okunus A.Genetic and geographic polymorphism of cultivated tobaccos(Nicotiana tabacum)in Turkey.Plant Genetics,2006,42:667-671.
[25]Julio E,Verrier J L,de Borne F D.Development of SCAR markers linked to three disease resistances based on AFLP within Nicotiana tabacum L.Theor Appl Genet,2006,112:335-346.
[26]Moon H S,Nicholson J S,Lewis R S.Use of transferable Nicotiana tabacum L.microsatellite markers for investigating genetic diversity in the genus Nicotiana.Genome,2008,51:547-559.
[27]Raju K S,Madhav M S,Sharma R K,Murthy T G K,Mohapatra T.Genetic polymorphism of Indian tobacco types as revealed by amplified fragment length polymorphism.Curr Sci,2008,94:633-638.
[28]Moon H S,Nicholson J S,Heineman A,Lion K,der Hoeven R V,Hayes A J,Lewis R S.Changes in genetic diversity of U.S.flue-cured tobacco germplasm over seven decades of cultivar development.Crop Sci,2009,49:498-506.
[29]Moon H S,Nifong J M,Nicholson J S,Heineman A,Lion K,der Hoeven R V,Hayes A J,Lewis R S.Microsatellite-based analysis of tobacco(Nicotiana tabacum L.)genetic resources.Crop Sci,2009,49:2149-2157.
[30]Bindler G,der Hoeven V R,Gunduz I,Plieske J,Ganal M,Rossi L,Gadani F,Donini P.A microsatellite marker based linkage map of tobacco.Theor Appl Genet,2007,114:341-349.
[31]Bindler G,Plieske J,Bakaher N,Gunduz I,Ivanov N,der Hoeven R V,Ganal M,Donini P.A high density genetic map of tobacco(Nicotiana tabacum L.)obtained from large scale microsatellite marker development.Theor Appl Genet,2011,123:219-230.
[32]Tong Z J,Yang Z M,Chen X J,Jiao F C,Li X Y,Wu X F,Gao YL,Xiao B G,Wu W R.Large-scale development of microsatellite markers in Nicotiana tabacum and construction of a genetic map of flue-cured tobacco.Plant Breed,2012,131:674-680.
[33]Tong Z J,Xiao B G,Jiao F C,Fang D H,Zeng J M,Wu X F,Chen X J,Yang J K,Li Y P.Large-scale development of SSRmarkers in tobacco and construction of a linkage map in flue-cured tobacco.Breed Sci,2016,66:381-390.
[34]郭永峰,朱贤朝,孔凡玉,石金开,王年.赤星病抗源的抗性比较.中国烟草科学,1997,11(4):1-6.Guo Y F,Zhu X C,Kong F Y,Shi J K,Wang N.Comparing two origins of resistance to brown spot disease in tobacco.Chin Tobacco Sci,1997,11(4):1-6(in Chinese with English abstract).
[35]许绍斌,陶玉芬,杨昭庆,褚嘉.简单快速的DNA银染和胶保存方法.遗传,2002,24:335-336.Xu S B,Tao Y F,Yang Z Q,Chu J.A simple and rapid methods used for silver staining and gel preservation.Hereditas,2002,24:335-336(in Chinese with English abstract).
[36]Van Ooijen J W.JoinMap?4.0,Software for the calculation of genetic linkage maps in experimental populations.Wageningen:Kyazma B.V Press,2006.pp 1-63.
[37]Voorrips R E.MapChart:Software for the graphical presentation of linkage maps and QTLs.J Heredity,2002,93:77-78.
[38]Van Ooijen J W.MapQTL?6.0,Software for the mapping of quantitative trait loci in experimental populations of diploid species.Wageningen:Kyazma B V Press,2009.pp 1-64.
[39]Mc Couch S R,Cho Y G,Yano M,Paul E,Blinstrub M,Morishima H,Kinoshita T.Report on QTL nomenclature.Rice Genet Newsl,1997,14:11-13.
[40]冯莹,蒋彩虹,程立锐,杨爱国,郑吉云,赵清海,杨修峰,尹华玲,冯全福.两个烟草赤星病抗源的遗传分析.中国烟草科学,2015,36(5):1-7.Feng Y,Jiang C H,Cheng L R,Yang A G,Zheng J Y,Zhao QH,Yang X F,Yin H L,Feng Q F.Genetic analysis of resistance to brown spot disease in tobacco cultivars Jingyehuang and Beinhart1000-1.Chin Tobacco Sci,2015,36(5):1-7(in Chinese with English abstract).