水稻粒宽基因GW5和GW8功能标记的开发与多重PCR检测
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摘要
水稻粒宽是衡量粒型、影响产量和品质的重要性状。本研究基于功能标记的开发,设计了水稻粒宽主效基因GW5和GW8的多重PCR检测体系,利用该体系对58份水稻新品系和品种进行鉴定。结果表明,GW5和GW8的多重PCR体系稳定、高效、重复性好。多重PCR检测发现粳稻品种津粳优2186和象牙香占等7个籼稻品种为GW5和GW8双插入型,平均粒宽2.32 mm;乐播1号和盐稻15198为GW8插入型,平均粒宽3.16 mm;其余48个品种为两基因缺失型,平均粒宽3.27 mm。研究结果为水稻粒型改良提供材料资源和分子标记检测技术手段。
Rice grain width is an important trait to measure grain size and closely related to yield and quality. In this study, a multiple PCR detection system for the main effect gene GW5 and GW8 of rice grain width was designed based on the development of functional markers. The system was used to identify 58 new lines and varieties of rice. The results showed that the multiple PCR system of GW5 and GW8 was stable, efficient and reproducible.The multiplex PCR detection found that japonica rice Jinjingyou2186 and 7 indica rice varieties such as Xiangyaxianzhan were GW5 and GW8 double inserted, and the average grain width was 2.32 mm. The Lebo No.1 and the Yandao15198 were the inserted type of GW8 with an average grain width of 3.16 mm. The other 48 varieties were two gene deletion types with an average grain width of 3.27 mm. The results could provide material resources and molecular marker assisted detection techniques for rice grain shape improvement.
Rice grain width is an important trait to measure grain size and closely related to yield and quality. In this study, a multiple PCR detection system for the main effect gene GW5 and GW8 of rice grain width was designed based on the development of functional markers. The system was used to identify 58 new lines and varieties of rice. The results showed that the multiple PCR system of GW5 and GW8 was stable, efficient and reproducible.The multiplex PCR detection found that japonica rice Jinjingyou2186 and 7 indica rice varieties such as Xiangyaxianzhan were GW5 and GW8 double inserted, and the average grain width was 2.32 mm. The Lebo No.1 and the Yandao15198 were the inserted type of GW8 with an average grain width of 3.16 mm. The other 48 varieties were two gene deletion types with an average grain width of 3.27 mm. The results could provide material resources and molecular marker assisted detection techniques for rice grain shape improvement.
引文
Dany H.,and Hidenori S.,2012a,An atypical bHLH protein encoded by positive regulator of grain length 2 is involved in controlling grain length and weight of rice through interaction with a typical bHLH protein APG,Breeding Science,62(2):133-141
Dany H.,and Hidenori S.,2012b,Antagonistic actions of HLH/bHLH proteins are involved in grain length and weight in rice,PLoS One,7(2):e31325
Fan C.C.,Xing Y.Z.,Mao H.L.,Lu T.T.,Han B.,Xu C.G.,Li X.H.,and Zhang Q.F.,2006,GS3,a major QTL for grain length and weight and minor QTL for grain width and thickness in rice,encodes a putative transmembrane protein,Theor.Appl.Genet.,112:1l64-1171
Fu F.H.,Wang F.,Huang J.W.,Peng H.P.,Wu Y.W.,and Huang D.J.,1994,Genetic analysis of grain traits in hybrid rice,Zuowu Xuebao(Acta Agronomica Sinica),20(1):39-45(符福鸿,王丰,黄文剑,彭惠普,伍应运,黄德娟,1994,杂交水稻谷粒性状的遗传分析,作物学报,20(1):39-45)
Huang K.,Wang D.K.,Duan P.G.,Zhang B.L.,Xu R.,Li N.,and Li Y.H.,2017,Wide and thick grain 1,which encodes an otubain-like protease with deubiquitylation activity,influences grain size and shape in rice,The Plant Journal,91(5):849-860
Huang R.Y.,Jiang L.R.,Zheng J.S.,Wang T.S.,Wang H.C.,Huang Y.M.,and Hong Z.L.,2013,Genetic bases of rice grain shape:so many genes,so little known,Trends Plant Sci.,18(4):218-226
Liu J.F.,Chen J.,Zheng X.M.,Wu F.Q.,Lin Q.B.,Heng Y.Q.,Tian P.,Cheng Z.J.,Yu X.W.,Zhou K.N.,Zhang X.,Guo X.P.,Wang J.L.,Wang H.Y.,and Wan J.M.,2017,GW5acts in the brassinosteroid signaling pathway to regulate grain width and weight in rice,Nature Plants,3:17043
Liu X.,Mou C.L.,Zhou C.L.,Cheng Z.J.,Jiang L.,and Wan J.M.,2018,Research progress on cloning and regulation mechanism of rice grain shape genes,Zhongguo Shuidao Kexue(Chinese Journal Rice Science),32(1):1-11(刘喜,牟昌铃,周春雷,程治军,江玲,万建民,2018,水稻粒型基因克隆和调控机制研究进展,中国水稻科学,32(1):1-11)
Onishi M.,Matsuoka T.,Kodama T.,Kashiwaba K.,Futo S.,Akiyama H.,Maitani T.,Furui S.,Oguchi T.,and Hino A.,2005,Development of a multiplex polymerase chain reaction method for simultaneous detection of eight events of genetically modified maize,Journal of Agricultural and Food Chemistry,53(25):9713-9721
Ribaut J.M.,and Hoisington D.,1998,Marker-assisted selection:new tools and strategies,Trends in Plant Science,3(6):236-239
Shao G.N.,Tang S.Q.,Luo J.,Jiao G.A.,Wei X.J.,Tang A.,Wu J.L.,Zhuang J.Y.,and Hu P.S.,2010,Mapping of qGL7-2,a grain length QTL on chromosome 7 of rice,J.Genet.Genomics,37(8):523-531
Shomura A.,Izawa T.,Ebana K.,Ebitani T.,Kanegae H.,Konishi S.,and Yano M.,2008,Deletion in a gene associated with grain size increased yields during rice domestication,Nature Genetics,40(8):1023-1028
Song X.J.,Huang W.,Shi M.,Zhu M.Z.,and Lin H.X.,2007,AQTL for rice grain width and weight encodes a previously unknown RING-type E3 ubiquitin ligase,Nature Genetics,39(5):623-630
Sun L.J.,Li X.J.,Fu Y.C.,Zhu Z.F.,Tan L.B.,Liu F.X.,Sun X.Y.,Sun X.W.,and Sun C.Q.,2013,GS6,a member of the GRAS gene family,negatively regulates grain size in rice,Journal of Integrative Plant Biology,55(10):938-949
Tapia R.,Quijano A.,Rojas R.,Larque A.,and Perez D.,2005,Afast,simple,and reliable high-yielding method for DNA extraction from different plant species,Molecular Biotechnology,31(2):137-139
Wang G.L.,Qiu Y.X.,and Fan H.Y.,2015,Establishment and application of PCR detection system of genetically modified maize in laboratory,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(8):1778-1783(王国莉,丘裕雄,范红英,2015,转基因玉米实验室PCR检测体系的优化及应用,基因组学与应用生物学,34(8):1778-1783)
Wang S.K.,Li S.,Liu Q.,Wu K.,Zhang J.Q.,Wang S.S.,Wang Y.,Chen X.B.,Zhang Y.,Gao C.X.,Wang F.,Huang H.X.,and Fu X.D.,2015,The OsSPL16-GW7 regulatory module determines grain shape and simultaneously improves rice yield and grain quality,Nature Genetics,47(8):949-954
Weng J.F.,Gu S.H.,Wan X.Y.,Gao H.,Guo T.,Su N.,Lei C.L.,Zhang X.,Cheng Z.J.,Guo X.P.,Wang J.L.,Jiang L.,Zhai H.Q.,and Wan J.M.,2008,Isolation and initial characterization of GW5,a major QTL associated with rice grain width and weight,Cell Research,18(12):1199-1209
Wu W.G.,Liu X.Y.,Wang M.H.,Rachel S.,Luo X.J.,Marie-Noelle N.,Tian L.B.,Zhang J.W.,Wu J.Z.,Cai H.W.,Sun C.Q.,Wang X.K.,Rod A.,and Zhu Z.F.,2017,A single-nucleotide polymorphism causes smaller grain size and loss of seed shattering during African rice domestication,Nature Plants,3:17064
Xing Y.Z.,and Zhang Q.F.,2010,Genetic and molecular bases of rice yield,Annu.Rev.Plant Biol.,61(1):421-442
Xu F.,Fang J.,Ou S.J.,Gao S.P.,Zhang F.X.,Du L.,Xiao Y.H.,Wang H.R.,Sun X.H.,Chu J.F.,Wang J.D.,and Chu C.C.,2015,Variations in CYP78A13 coding region influence grain size and yield in rice,Plant Cell&Environment,38(4):800-811
Yang J.,Cao Q.,Wang J.,Fan F.J.,Zhang Y.Q.,and Zhong W.G.,2011,Development and application of functional markers for rice polyphenol oxidase gene,Zhongguo Shuidao Kexue(Rice Science in China),25(1):37-42(杨杰,曹卿,王军,范方军,张玉琼,仲维功,2011,水稻多酚氧化酶基因功能标记的开发与应用,中国水稻科学,25(1):37-42)
Dany H.,and Hidenori S.,2012b,Antagonistic actions of HLH/bHLH proteins are involved in grain length and weight in rice,PLoS One,7(2):e31325
Fan C.C.,Xing Y.Z.,Mao H.L.,Lu T.T.,Han B.,Xu C.G.,Li X.H.,and Zhang Q.F.,2006,GS3,a major QTL for grain length and weight and minor QTL for grain width and thickness in rice,encodes a putative transmembrane protein,Theor.Appl.Genet.,112:1l64-1171
Fu F.H.,Wang F.,Huang J.W.,Peng H.P.,Wu Y.W.,and Huang D.J.,1994,Genetic analysis of grain traits in hybrid rice,Zuowu Xuebao(Acta Agronomica Sinica),20(1):39-45(符福鸿,王丰,黄文剑,彭惠普,伍应运,黄德娟,1994,杂交水稻谷粒性状的遗传分析,作物学报,20(1):39-45)
Huang K.,Wang D.K.,Duan P.G.,Zhang B.L.,Xu R.,Li N.,and Li Y.H.,2017,Wide and thick grain 1,which encodes an otubain-like protease with deubiquitylation activity,influences grain size and shape in rice,The Plant Journal,91(5):849-860
Huang R.Y.,Jiang L.R.,Zheng J.S.,Wang T.S.,Wang H.C.,Huang Y.M.,and Hong Z.L.,2013,Genetic bases of rice grain shape:so many genes,so little known,Trends Plant Sci.,18(4):218-226
Liu J.F.,Chen J.,Zheng X.M.,Wu F.Q.,Lin Q.B.,Heng Y.Q.,Tian P.,Cheng Z.J.,Yu X.W.,Zhou K.N.,Zhang X.,Guo X.P.,Wang J.L.,Wang H.Y.,and Wan J.M.,2017,GW5acts in the brassinosteroid signaling pathway to regulate grain width and weight in rice,Nature Plants,3:17043
Liu X.,Mou C.L.,Zhou C.L.,Cheng Z.J.,Jiang L.,and Wan J.M.,2018,Research progress on cloning and regulation mechanism of rice grain shape genes,Zhongguo Shuidao Kexue(Chinese Journal Rice Science),32(1):1-11(刘喜,牟昌铃,周春雷,程治军,江玲,万建民,2018,水稻粒型基因克隆和调控机制研究进展,中国水稻科学,32(1):1-11)
Onishi M.,Matsuoka T.,Kodama T.,Kashiwaba K.,Futo S.,Akiyama H.,Maitani T.,Furui S.,Oguchi T.,and Hino A.,2005,Development of a multiplex polymerase chain reaction method for simultaneous detection of eight events of genetically modified maize,Journal of Agricultural and Food Chemistry,53(25):9713-9721
Ribaut J.M.,and Hoisington D.,1998,Marker-assisted selection:new tools and strategies,Trends in Plant Science,3(6):236-239
Shao G.N.,Tang S.Q.,Luo J.,Jiao G.A.,Wei X.J.,Tang A.,Wu J.L.,Zhuang J.Y.,and Hu P.S.,2010,Mapping of qGL7-2,a grain length QTL on chromosome 7 of rice,J.Genet.Genomics,37(8):523-531
Shomura A.,Izawa T.,Ebana K.,Ebitani T.,Kanegae H.,Konishi S.,and Yano M.,2008,Deletion in a gene associated with grain size increased yields during rice domestication,Nature Genetics,40(8):1023-1028
Song X.J.,Huang W.,Shi M.,Zhu M.Z.,and Lin H.X.,2007,AQTL for rice grain width and weight encodes a previously unknown RING-type E3 ubiquitin ligase,Nature Genetics,39(5):623-630
Sun L.J.,Li X.J.,Fu Y.C.,Zhu Z.F.,Tan L.B.,Liu F.X.,Sun X.Y.,Sun X.W.,and Sun C.Q.,2013,GS6,a member of the GRAS gene family,negatively regulates grain size in rice,Journal of Integrative Plant Biology,55(10):938-949
Tapia R.,Quijano A.,Rojas R.,Larque A.,and Perez D.,2005,Afast,simple,and reliable high-yielding method for DNA extraction from different plant species,Molecular Biotechnology,31(2):137-139
Wang G.L.,Qiu Y.X.,and Fan H.Y.,2015,Establishment and application of PCR detection system of genetically modified maize in laboratory,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(8):1778-1783(王国莉,丘裕雄,范红英,2015,转基因玉米实验室PCR检测体系的优化及应用,基因组学与应用生物学,34(8):1778-1783)
Wang S.K.,Li S.,Liu Q.,Wu K.,Zhang J.Q.,Wang S.S.,Wang Y.,Chen X.B.,Zhang Y.,Gao C.X.,Wang F.,Huang H.X.,and Fu X.D.,2015,The OsSPL16-GW7 regulatory module determines grain shape and simultaneously improves rice yield and grain quality,Nature Genetics,47(8):949-954
Weng J.F.,Gu S.H.,Wan X.Y.,Gao H.,Guo T.,Su N.,Lei C.L.,Zhang X.,Cheng Z.J.,Guo X.P.,Wang J.L.,Jiang L.,Zhai H.Q.,and Wan J.M.,2008,Isolation and initial characterization of GW5,a major QTL associated with rice grain width and weight,Cell Research,18(12):1199-1209
Wu W.G.,Liu X.Y.,Wang M.H.,Rachel S.,Luo X.J.,Marie-Noelle N.,Tian L.B.,Zhang J.W.,Wu J.Z.,Cai H.W.,Sun C.Q.,Wang X.K.,Rod A.,and Zhu Z.F.,2017,A single-nucleotide polymorphism causes smaller grain size and loss of seed shattering during African rice domestication,Nature Plants,3:17064
Xing Y.Z.,and Zhang Q.F.,2010,Genetic and molecular bases of rice yield,Annu.Rev.Plant Biol.,61(1):421-442
Xu F.,Fang J.,Ou S.J.,Gao S.P.,Zhang F.X.,Du L.,Xiao Y.H.,Wang H.R.,Sun X.H.,Chu J.F.,Wang J.D.,and Chu C.C.,2015,Variations in CYP78A13 coding region influence grain size and yield in rice,Plant Cell&Environment,38(4):800-811
Yang J.,Cao Q.,Wang J.,Fan F.J.,Zhang Y.Q.,and Zhong W.G.,2011,Development and application of functional markers for rice polyphenol oxidase gene,Zhongguo Shuidao Kexue(Rice Science in China),25(1):37-42(杨杰,曹卿,王军,范方军,张玉琼,仲维功,2011,水稻多酚氧化酶基因功能标记的开发与应用,中国水稻科学,25(1):37-42)