甜瓜MR-1细菌性果斑病抗病基因QTL定位分析
|
摘要
为了研究甜瓜细菌性果斑病的抗病基因,以甜瓜(Cucumis melo L.)抗细菌性果斑病品种‘MR-1’和感果斑病品种‘伽师瓜’为亲本构建F2群体,研究‘MR-1’中细菌性果斑病抗性遗传规律及基因定位。群体遗传分析表明,F2分离群体病情级别个数呈现连续分布,为多基因控制。采用简单重复序列(simple sequence repeats, SSR)分子标记结合集团分离分析法(bulked segregation analysis, BSA)对‘MR-1’抗果斑病位点进行初步定位。利用全基因组范围内288个SSR分子标记检测抗果斑病、感果斑病基因池,筛选出15个具有多态性的标记分别位于LG5、LG6、LG12。对个体进行SSR分子标记基因型检测和连锁分析,将果斑病抗性数量性状位点(quantitative trait locus, QTL)初步定位于LG12。利用针对LG12上设计的共90对SSR引物检测‘MR-1’和‘伽师瓜’,其中共20个SSR标记在双亲之间表现为多态性。利用具有多态的分子标记构建LG12的遗传连锁图,进一步将bfb-QTL12.1定位于CMMS35-4与DE1851之间,两标记间物理距离约3.4 Mb,遗传距离为9.1 cM。本研究为后续利用抗性品种进行品种改良提供理论依据。
In order to study the resistance genes of melon bacterial fruit blotch, this study took the antibacterial fruit blotch breed 'MR-1' and the susceptible melon 'Jiashigua'as parent to construct in F2 population and study the genetic regularity and gene location of bacterial fruit blotch resistence in 'MR-1'. Genetic analysis showed that the segregation data of F2 population was continuous distribution and controlled by multiple genes. This study primary located the locus of resisit fruit blotch by Simple sequence repeats(SSR) markers combined with Bulked Segregation Analysis(BSA) method. Among 288 gene pools for detecting the resistant bacterial fruit blotch and feeling bacterial fruit blotch by SSR markers,we detected 15 polymorphic markers, which were located at LG5, LG6 and LG12 respectively. By genotyping these individuals with SSR markers and performing linkage analysis showed that the major resistance QTL was located on LG12. A total of 90 SSR primers on LG12 were tested to identify the'MR-1' and 'Jiashigua'. A linkage map was developed constructed using 20 SSR markers with polymorphism between the parents. We constructed the genetic linkage map of LG12 by molecular maker with polymorphic. And the bfb-QTL12.1 was located between CMMS35-4 and DE1851 with physical distance of 3.4 Mb and genetic distance of 9.1 cM. This study might lay the foundation for the subsequent improvement of varieties using resistant varieties.
In order to study the resistance genes of melon bacterial fruit blotch, this study took the antibacterial fruit blotch breed 'MR-1' and the susceptible melon 'Jiashigua'as parent to construct in F2 population and study the genetic regularity and gene location of bacterial fruit blotch resistence in 'MR-1'. Genetic analysis showed that the segregation data of F2 population was continuous distribution and controlled by multiple genes. This study primary located the locus of resisit fruit blotch by Simple sequence repeats(SSR) markers combined with Bulked Segregation Analysis(BSA) method. Among 288 gene pools for detecting the resistant bacterial fruit blotch and feeling bacterial fruit blotch by SSR markers,we detected 15 polymorphic markers, which were located at LG5, LG6 and LG12 respectively. By genotyping these individuals with SSR markers and performing linkage analysis showed that the major resistance QTL was located on LG12. A total of 90 SSR primers on LG12 were tested to identify the'MR-1' and 'Jiashigua'. A linkage map was developed constructed using 20 SSR markers with polymorphism between the parents. We constructed the genetic linkage map of LG12 by molecular maker with polymorphic. And the bfb-QTL12.1 was located between CMMS35-4 and DE1851 with physical distance of 3.4 Mb and genetic distance of 9.1 cM. This study might lay the foundation for the subsequent improvement of varieties using resistant varieties.
引文
Bahar O.,Burdman S.,Edelstein M.,and Cohen R.,2010,Bacterial fruit blotch:A threat to the cucurbit industry,Israel Journal of Plant Sciences,58(1):19-31
Bahar O.,Kritzman G.,and Burdman S.,2009,Bacterial fruit blotch of melon:screens for disease tolerance and role of seed transmission in pathogenicity,Eur.J.Plant Pathol.,123(1):71-83
Chen Y.P.,Meng G.Q.,and Yuan J.H.,2008,Identification of the molecular markers linked to the MRDV-resistance locus in maize using SSR-BSA technique,Jiangsu Nongye Kexue(Jiangsu Agricultural Sciences),24(5):590-594(陈艳萍,孟庆长,袁建华,2008,利用SSR-BSA技术筛选玉米粗缩病抗性基因分子标记,江苏农业学报,24(5):590-594)
Fukino N.,Ohara T.,Sugiyama M.,Kubo N.,Hirai M.,Sakata Y.,and Matsumoto S.,2012,Mapping of a gene that confers short lateral branching(slb)in melon(Cucumis melo L.),E-uphytica,187(1):145-146
Garcia-Mas J.,Benjak A.,Sanseverino W.,Bourgeois M.,Mir G.,González V.M.,Hénaff E.,C覾mara F.,Cozzuto L.,Lowy E.,Alioto T.,Capella-Gutiérrez S.,Blanca J.,Ca觡izares J.,Ziarsolo P.,Gonzalez-Ibeas D.,Rodríguez-Moreno L.,Droege M.,Du L.,Alvarez-Tejado M.,Lorente-Galdos B.,MeléM.,Yang L.,Weng Y.,Navarro A.,Marques-Bonet T.,Aranda M.A.,Nuez F.,PicóB.,Gabaldón T.,Roma G.,GuigóR.,Casacuberta J.M.,Arús P.,and Puigdomènech P.,2012,The genome of melon(Cucumis melo L.),Proc.Natl.Acad.Sci.USA,109(29):11872-11877
Gao X.W.,Ning X.F.,Wang Y.M.,Wang X.L.,Yan W.L.,Zhang Z.Q.,and Guan L.,2014,Fine mapping of a gene that confers palmately lobed leaf(pll)in melon(Cucumis melo L.),Euphytica,200(3):337-347
Hopkins D.L.,and Thompson C.M.,2002,Evaluation of Citrullus sp.germ plasm for resistance to Acidovorax avenae subsp.Citrulli,Plant Disease,86(1):61-64
Hwang J.,Oh J.,Kim Z.,Staub J.E.,Chung S.M.,and Par Y.H.,2014,Fine genetic mapping of a locus controlling short internode length in melon(Cucumis melo L.),Mol.Breeding,34(3):949-961
Han Q.D.,Chen Z.W.,Deng Y.,Lan T.,Guan H.Z.,Duan Y.L.,Zhou Y.C.,Lin M.C.,and Wu W.R.,2008,Fine mapping of qBlsr5a,a QTL controlling resistance to bacterial leaf streak in rice,Zuowu Xuebao(Acta Agronomica Sinica),34(4):587-590(韩庆典,陈志伟,邓云,兰涛,官华忠,段远霖,周元昌,林闽川,吴为人,2008,水稻细菌性条斑病抗性QTLqB1sr5a的精细定位,作物学报,34(4):587-590)
Hui W.G.,Zhao T.C.,Schaad N.W.,Sun F.Z.,and Wang J.R.,2007,Establishment of rapid detection method for the pathogen of hami melon fruit blotch,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),40(11):2495-2501(回文广,赵廷昌,Schaad N.W.,孙福在,王建荣,2007,哈密瓜细菌性果斑病菌快速检测方法的建立,中国农业科学,40(11):2495-2501)
Li J.G.,2015,Resistance identification of different muskmelons to Acidovorax citrulli and studies on mechanism of disease resistance,Thesis for M.S.,Xinjiang Agricultural University,Supervisor:Wang H.L.,pp.1-53(李俊阁,2015,甜瓜细菌性果斑病抗性鉴定及抗性相关生理研究,硕士学位论文,新疆农业大学,导师:王惠林,pp.1-53)
Li J.G.,Wang H.L.,Zhang L.,Zheng J.,and Gao Q.,2015,Identification of resistance to Acidovorax avenae subsp.Citrulli among different melons(Cucumis spp.)at the seeding stage,Xinjing Nongye Kexue(Xinjiang Agricultural Sciences),52(10):1843-1848(李俊阁,王惠林,张亮,郑健,高强,2015,不同甜瓜材料苗期对细菌性果斑病抗病性鉴定,新疆农业科学,52(10):1843-184)
Li W.,Ren S.Z.,Jing J.J.,Hou P.,and Xie F.S.,2007,Identification of citrull resistance to Acidovorax avenae subsp.Citrulli in Xinjiang,Beifang Yuanyi(Northern Horticulture),(3):186-188(李威,任毓忠,丁建军,侯鹏,谢方生,2007,新疆瓜类细菌性果斑病品种抗病性鉴定,北方园艺,(3):186-188)
Lu H.,Wang X.L.,Gao X.W.,Ning X.F.,Chen J.,and Li G.,2015,Inheritance and location of powdery mildew resistance gene in melon'PMR6',Yuanyi Xuebao(Acta Horticulturae Sinica),42(6):1121-1128(卢浩,王贤磊,高兴旺,宁雪飞,陈静,李冠,2015,甜瓜'PMR6'抗白粉病基因的遗传及其定位研究,园艺学报,42(6):1121-1128)
Luo R.,Wu W.L.,Zhang Y.,and Li Y.H.,2010,SSR marker and its application to crop genetics and breeding,Jiyinzuxue yu Yingyong Shengwuxue(Genomics and Applied Biology),29(1):137-143(罗冉,吴委林,张旸,李玉花,2010,SSR分子标记在作物遗传育种中的应用,基因组学与应用生物学,29(1):137-143)
Mao L.,Ji Y.L.,Zhang J.,Wang C.X.,Pan W.P.,Zhang Y.H.,and Wang X.D.,2017,A preliminary study on resistance of hami melon varieties to Acidovorax avenae subsp,citrulli in turpan area,Xinjing Nongye Kexue(Xinjiang Agricultural Sciences),54(11):2054-2059(毛亮,吉艳玲,张娟,王宣仓,潘卫萍,张以和,王晓东,2017,吐鲁番地区哈密瓜品种对细菌性果斑病抗性研究,新疆农业科学,54(11):2054-2059)
Ning X.F.,Wang X.L.,Gao X.W.,Zhang Z.Q.,Zhang L.H.,Yan W.L.,and Guan L.,2014,Inheritances and location of powdery mildew resistance gene in melon edisto47,Euphytica,195(3):345-353
Pu Y.Y.,Cheng K.,and Li S.S.,2011,Genetic diversity of recent wheat cultivars in Shandong province using SSR markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),9(4):443-449(蒲艳艳,程凯,李斯深,2011,山东省近期育成小麦品种遗传多样性的SSR分析,分子植物育种,9(4):443-449)
Wang R.Q.,Zhao P.,Wang D.D.,Wang Y.H.,and Chen Q.,2018,Genetic relationship analysis and finger print profile construction of pigmented potato based on SSR markers,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),38(2):249-257(王若秋,赵朋,王冬冬,王耀红,陈勤,2018,基于SSR标记的彩色马铃薯亲缘关系分析及指纹图谱构建,38(2):249-257)
Wang X.L.,Li G.,Gao X.,Xiong L.W.,Xiong L.M.,Wang W.,and Han R.,2011,Powdery mildew resistance gene(Pm-AN)located in a segregation distortion region of melon LGV,Euphytica,180(3):421-428
Walcott R.R.,ed.,2008,Integrated pest management of bacterial fruit blotch cucurbits,Berlin,Germany,Springer,pp.187-205
Wang X.H.,Liu X.Y.,and Li G.,2015,Isolation,Isolation,identification and 16S r DNA sequence analysis of bacterial fruit blotch pathogen in melon,Shiping Kexue(Food Science),36(19):186-189(王新绘,刘晓颖,李冠,2015,甜瓜细菌性果斑病病原菌的分离鉴定及16S r DNA序列分析,食品科学,36(19):186-189)
Yang X.L.,Ma J.Y.,Hu B.S.,Chen T.,and Li X.X.,2008,Identification of disease resistance of different varieties of hami melon at seedling stage to Acidovorax avenae subsp.citrulli,Xinjing Nongye Kexue(Xinjiang Agricultural Sciences),45(3):452-455(杨小丽,马俊义,胡白石,陈涛,李晓霞,2008,不同哈密瓜品种苗期对细菌性果斑病的抗病性鉴定,新疆农业科学,45(3):452-455)
Zhang C.Q.,Ren Y.,Guo S.G.,Zhang H.Y.,Gong G.Y.,Du Y.C,and Xu Y.,2013,Application of comparative genomics in developing markers tightly linked to the Pm-2F gene for powdery mildew resistance in melon(Cucumis melo L.),Euphytica,190(2):157-168
Zhang L.,2016,Resistance identification of different melons to Acidovorax citrulli and studies on morph-physiology of disease resistance,Thesis for M.S.,Xinjiang Agricultural University,Supervisor:Wang H.L.,pp.1-53(张亮,2016,甜瓜果斑病抗性鉴定及抗性相关形态和生理研究,硕士学位论文,新疆农业大学,导师:王惠林,pp.1-45)
Zhang L.,Wang H.L.,and Wan X.Q.,2016,Identification of resistance to Acidovorax citrulli of different melons in the seeding stage,Beifang Yuanyi(Northern Horticulture),(9):117-120(张亮,王惠林,万秀琴,2016,不同甜瓜材料苗期细菌性果斑病抗性鉴定,北方园艺,(9):117-120)
Bahar O.,Kritzman G.,and Burdman S.,2009,Bacterial fruit blotch of melon:screens for disease tolerance and role of seed transmission in pathogenicity,Eur.J.Plant Pathol.,123(1):71-83
Chen Y.P.,Meng G.Q.,and Yuan J.H.,2008,Identification of the molecular markers linked to the MRDV-resistance locus in maize using SSR-BSA technique,Jiangsu Nongye Kexue(Jiangsu Agricultural Sciences),24(5):590-594(陈艳萍,孟庆长,袁建华,2008,利用SSR-BSA技术筛选玉米粗缩病抗性基因分子标记,江苏农业学报,24(5):590-594)
Fukino N.,Ohara T.,Sugiyama M.,Kubo N.,Hirai M.,Sakata Y.,and Matsumoto S.,2012,Mapping of a gene that confers short lateral branching(slb)in melon(Cucumis melo L.),E-uphytica,187(1):145-146
Garcia-Mas J.,Benjak A.,Sanseverino W.,Bourgeois M.,Mir G.,González V.M.,Hénaff E.,C覾mara F.,Cozzuto L.,Lowy E.,Alioto T.,Capella-Gutiérrez S.,Blanca J.,Ca觡izares J.,Ziarsolo P.,Gonzalez-Ibeas D.,Rodríguez-Moreno L.,Droege M.,Du L.,Alvarez-Tejado M.,Lorente-Galdos B.,MeléM.,Yang L.,Weng Y.,Navarro A.,Marques-Bonet T.,Aranda M.A.,Nuez F.,PicóB.,Gabaldón T.,Roma G.,GuigóR.,Casacuberta J.M.,Arús P.,and Puigdomènech P.,2012,The genome of melon(Cucumis melo L.),Proc.Natl.Acad.Sci.USA,109(29):11872-11877
Gao X.W.,Ning X.F.,Wang Y.M.,Wang X.L.,Yan W.L.,Zhang Z.Q.,and Guan L.,2014,Fine mapping of a gene that confers palmately lobed leaf(pll)in melon(Cucumis melo L.),Euphytica,200(3):337-347
Hopkins D.L.,and Thompson C.M.,2002,Evaluation of Citrullus sp.germ plasm for resistance to Acidovorax avenae subsp.Citrulli,Plant Disease,86(1):61-64
Hwang J.,Oh J.,Kim Z.,Staub J.E.,Chung S.M.,and Par Y.H.,2014,Fine genetic mapping of a locus controlling short internode length in melon(Cucumis melo L.),Mol.Breeding,34(3):949-961
Han Q.D.,Chen Z.W.,Deng Y.,Lan T.,Guan H.Z.,Duan Y.L.,Zhou Y.C.,Lin M.C.,and Wu W.R.,2008,Fine mapping of qBlsr5a,a QTL controlling resistance to bacterial leaf streak in rice,Zuowu Xuebao(Acta Agronomica Sinica),34(4):587-590(韩庆典,陈志伟,邓云,兰涛,官华忠,段远霖,周元昌,林闽川,吴为人,2008,水稻细菌性条斑病抗性QTLqB1sr5a的精细定位,作物学报,34(4):587-590)
Hui W.G.,Zhao T.C.,Schaad N.W.,Sun F.Z.,and Wang J.R.,2007,Establishment of rapid detection method for the pathogen of hami melon fruit blotch,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),40(11):2495-2501(回文广,赵廷昌,Schaad N.W.,孙福在,王建荣,2007,哈密瓜细菌性果斑病菌快速检测方法的建立,中国农业科学,40(11):2495-2501)
Li J.G.,2015,Resistance identification of different muskmelons to Acidovorax citrulli and studies on mechanism of disease resistance,Thesis for M.S.,Xinjiang Agricultural University,Supervisor:Wang H.L.,pp.1-53(李俊阁,2015,甜瓜细菌性果斑病抗性鉴定及抗性相关生理研究,硕士学位论文,新疆农业大学,导师:王惠林,pp.1-53)
Li J.G.,Wang H.L.,Zhang L.,Zheng J.,and Gao Q.,2015,Identification of resistance to Acidovorax avenae subsp.Citrulli among different melons(Cucumis spp.)at the seeding stage,Xinjing Nongye Kexue(Xinjiang Agricultural Sciences),52(10):1843-1848(李俊阁,王惠林,张亮,郑健,高强,2015,不同甜瓜材料苗期对细菌性果斑病抗病性鉴定,新疆农业科学,52(10):1843-184)
Li W.,Ren S.Z.,Jing J.J.,Hou P.,and Xie F.S.,2007,Identification of citrull resistance to Acidovorax avenae subsp.Citrulli in Xinjiang,Beifang Yuanyi(Northern Horticulture),(3):186-188(李威,任毓忠,丁建军,侯鹏,谢方生,2007,新疆瓜类细菌性果斑病品种抗病性鉴定,北方园艺,(3):186-188)
Lu H.,Wang X.L.,Gao X.W.,Ning X.F.,Chen J.,and Li G.,2015,Inheritance and location of powdery mildew resistance gene in melon'PMR6',Yuanyi Xuebao(Acta Horticulturae Sinica),42(6):1121-1128(卢浩,王贤磊,高兴旺,宁雪飞,陈静,李冠,2015,甜瓜'PMR6'抗白粉病基因的遗传及其定位研究,园艺学报,42(6):1121-1128)
Luo R.,Wu W.L.,Zhang Y.,and Li Y.H.,2010,SSR marker and its application to crop genetics and breeding,Jiyinzuxue yu Yingyong Shengwuxue(Genomics and Applied Biology),29(1):137-143(罗冉,吴委林,张旸,李玉花,2010,SSR分子标记在作物遗传育种中的应用,基因组学与应用生物学,29(1):137-143)
Mao L.,Ji Y.L.,Zhang J.,Wang C.X.,Pan W.P.,Zhang Y.H.,and Wang X.D.,2017,A preliminary study on resistance of hami melon varieties to Acidovorax avenae subsp,citrulli in turpan area,Xinjing Nongye Kexue(Xinjiang Agricultural Sciences),54(11):2054-2059(毛亮,吉艳玲,张娟,王宣仓,潘卫萍,张以和,王晓东,2017,吐鲁番地区哈密瓜品种对细菌性果斑病抗性研究,新疆农业科学,54(11):2054-2059)
Ning X.F.,Wang X.L.,Gao X.W.,Zhang Z.Q.,Zhang L.H.,Yan W.L.,and Guan L.,2014,Inheritances and location of powdery mildew resistance gene in melon edisto47,Euphytica,195(3):345-353
Pu Y.Y.,Cheng K.,and Li S.S.,2011,Genetic diversity of recent wheat cultivars in Shandong province using SSR markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),9(4):443-449(蒲艳艳,程凯,李斯深,2011,山东省近期育成小麦品种遗传多样性的SSR分析,分子植物育种,9(4):443-449)
Wang R.Q.,Zhao P.,Wang D.D.,Wang Y.H.,and Chen Q.,2018,Genetic relationship analysis and finger print profile construction of pigmented potato based on SSR markers,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),38(2):249-257(王若秋,赵朋,王冬冬,王耀红,陈勤,2018,基于SSR标记的彩色马铃薯亲缘关系分析及指纹图谱构建,38(2):249-257)
Wang X.L.,Li G.,Gao X.,Xiong L.W.,Xiong L.M.,Wang W.,and Han R.,2011,Powdery mildew resistance gene(Pm-AN)located in a segregation distortion region of melon LGV,Euphytica,180(3):421-428
Walcott R.R.,ed.,2008,Integrated pest management of bacterial fruit blotch cucurbits,Berlin,Germany,Springer,pp.187-205
Wang X.H.,Liu X.Y.,and Li G.,2015,Isolation,Isolation,identification and 16S r DNA sequence analysis of bacterial fruit blotch pathogen in melon,Shiping Kexue(Food Science),36(19):186-189(王新绘,刘晓颖,李冠,2015,甜瓜细菌性果斑病病原菌的分离鉴定及16S r DNA序列分析,食品科学,36(19):186-189)
Yang X.L.,Ma J.Y.,Hu B.S.,Chen T.,and Li X.X.,2008,Identification of disease resistance of different varieties of hami melon at seedling stage to Acidovorax avenae subsp.citrulli,Xinjing Nongye Kexue(Xinjiang Agricultural Sciences),45(3):452-455(杨小丽,马俊义,胡白石,陈涛,李晓霞,2008,不同哈密瓜品种苗期对细菌性果斑病的抗病性鉴定,新疆农业科学,45(3):452-455)
Zhang C.Q.,Ren Y.,Guo S.G.,Zhang H.Y.,Gong G.Y.,Du Y.C,and Xu Y.,2013,Application of comparative genomics in developing markers tightly linked to the Pm-2F gene for powdery mildew resistance in melon(Cucumis melo L.),Euphytica,190(2):157-168
Zhang L.,2016,Resistance identification of different melons to Acidovorax citrulli and studies on morph-physiology of disease resistance,Thesis for M.S.,Xinjiang Agricultural University,Supervisor:Wang H.L.,pp.1-53(张亮,2016,甜瓜果斑病抗性鉴定及抗性相关形态和生理研究,硕士学位论文,新疆农业大学,导师:王惠林,pp.1-45)
Zhang L.,Wang H.L.,and Wan X.Q.,2016,Identification of resistance to Acidovorax citrulli of different melons in the seeding stage,Beifang Yuanyi(Northern Horticulture),(9):117-120(张亮,王惠林,万秀琴,2016,不同甜瓜材料苗期细菌性果斑病抗性鉴定,北方园艺,(9):117-120)
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |