香稻材料Badh2基因第7外显子的序列突变及其香味鉴定
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摘要
本研究对12个香稻材料Badh2基因第7外显子序列进行PCR扩增和序列分析,利用籽粒咀嚼法、KOH浸泡法对香稻材料进行香味鉴定,以分析香稻材料Badh2基因第7外显子的序列突变及其与香味性状的对应关系。结果表明,香糯、绿金香、美香占2号、华香、越香选1号、红芋香、美香选1号、巴香选1号、泰香选1号、泰香选2号10个材料Badh2基因的第7外显子均发生了8 bp的缺失(aaaa--t——ggc)和1 bp的突变(g→t),与GenBank的EU155083序列的突变一致,在中国水稻材料中首次发现该突变。在发生突变的10个香稻材料中,7个材料被鉴定为香稻品种,其中美香占2号和越香选1号的糙米有甜味;华香和美香选1号被鉴定为杂合基因型,红芋香被鉴定为非香稻。香稻材料Badh2基因第7外显子的序列突变与其香味性状不能一一对应。研究结果为水稻香味性状遗传机制研究和香稻品种的推广提供了参考。
In this study, we performed the amplificaion and the sequence analysis of exon7 of Badh2 gene of 12 fragrant rice materials, and identified the aroma of fragrant rice materials by the method of seed chewing and KOH soaking, to analysed the sequence mutation of exon 7 in the Badh2 gene of rice material and its corresponding relation with the flavor character. The results showed that an 8 bp deletion(aaaa--t---ggc) and a mutation of SNP(g →t) in exon 7 of Badh2 gene were found in 10 materials, including Xiangnuo, Lüjinxiang, Meixiangzhan 2,Huaxiang, Yuexiangxuan 1, Hongyuxiang, Meixiangxuan 1, Baxiangxuan 1, Taixiangxuan 1, Taixiangxuan 2.This mutaion was consistent with the mutation of EU155083 sequence in GenBank and was reported for the first time in Chinese rice materials. In these 10 fragrant rice materials with mutation, 7 rice materials were identified as fragrant rice where the brown rice of Meixiangzhan 2 and Yuexiangyuan 1 had sweet taste, Huaxiang and Meixiangxuan 1 were identified as the heterozygote genotype, and Hongyuxiang was identified as the non-fragrant rice. So the sequence mutation in exon 7 of Badh2 gene in fragrant rice materials didn't correspond to aroma traits one by one. Our results could provide a reference for the research on the genetic mechanism of rice aroma character and the promotion of the rice variety.
In this study, we performed the amplificaion and the sequence analysis of exon7 of Badh2 gene of 12 fragrant rice materials, and identified the aroma of fragrant rice materials by the method of seed chewing and KOH soaking, to analysed the sequence mutation of exon 7 in the Badh2 gene of rice material and its corresponding relation with the flavor character. The results showed that an 8 bp deletion(aaaa--t---ggc) and a mutation of SNP(g →t) in exon 7 of Badh2 gene were found in 10 materials, including Xiangnuo, Lüjinxiang, Meixiangzhan 2,Huaxiang, Yuexiangxuan 1, Hongyuxiang, Meixiangxuan 1, Baxiangxuan 1, Taixiangxuan 1, Taixiangxuan 2.This mutaion was consistent with the mutation of EU155083 sequence in GenBank and was reported for the first time in Chinese rice materials. In these 10 fragrant rice materials with mutation, 7 rice materials were identified as fragrant rice where the brown rice of Meixiangzhan 2 and Yuexiangyuan 1 had sweet taste, Huaxiang and Meixiangxuan 1 were identified as the heterozygote genotype, and Hongyuxiang was identified as the non-fragrant rice. So the sequence mutation in exon 7 of Badh2 gene in fragrant rice materials didn't correspond to aroma traits one by one. Our results could provide a reference for the research on the genetic mechanism of rice aroma character and the promotion of the rice variety.
引文
Amarawathi Y.,Singh R.,Singh A.K.,Singh V.P.,Mohapatra T.,Sharma T.R.,and Singh N.K.,2008,Mapping of quantitative trait loci for basmati quality traits in rice(Oryza sativa L.),Mol.Breeding,21(1):49-65
Bindusree G.,Natarajan P.,Kalva S.,and Madasamy P.,2017,Whole genome sequencing of Oryza sativa L.cv.Seeragasamba identifies a new fragrance allele in rice,PLoS One,12(11):e0188920
Bourgis F.,Guyot R.,Gherbi H.,Tailliez E.,Amabile I.,Salse J.,Lorieux M.,Delseny M.,and Ghesquière A.,2008,Characterization of the major fragance gene from an aromatic japonica rice and analysis of its diversity in Asian cultivated rice,Theor.Appl.Genet.,117(3):353-368
Bradbury L.M.T.,Fitzgerald T.L.,Henry R.J.,Jin Q.S.,and Waters D.L.E.,2005,The gene for fragrance in rice,Plant Biotechnol.J.,3(3):363-370
Fitzgerald M.A.,Hamilton N.R.S.,Calingacion M.N.,Verhoeven H.A.,and Butardo V.M.,2008,Is there a second fragrance gene in rice?Plant Biotechnol.J.,6(4):416-423
Jiang D.,Lu X.Y.,Wang Y.C.,Fan Y.L.,Zhai W.X.,Luo Y.H.,and Xia Z.H.,2015,Sequence analysis of mutation site of Badh2 in 27 varieties of fragrant rice,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(2):276-280(姜达,卢小勇,王延春,范玉龙,翟文学,罗越华,夏志辉,2015,27种香稻品种badh2突变位点序列的分析,分子植物育种,13(2):276-280)
Kovach M.J.,Calingacionb M.N.,Fitzgeraldb M.A.,and McCoucha S.R.,2009,The origin and evolution of fragrance in rice(Oryza sativa L.),Proc.Natl.Acad.Sci.USA,106(34):14444-14449
Myint K.M.,Courtois B.,Risterucci A.M.,Frouin J.,Soe K.,Thet K.M.,Vanavichit A.,and Glaszmann J.C.,2012,Specific patterns of genetic diversity among aromatic rice varieties in Myanmar,Rice,5(1):1-13
Ootsuka K.,Takahashi I.,Tanaka K.,Itani T.,Tabuchi H.,Yoshihashi T.,Tonouchi A.,and Ishikawa R.,2014,Genetic polymorphisms in Japanese fragrant landraces and novel fragrant alleledomesticated in northern Japan,Breeding Science,64(2):115-124
Ren G.J.,Li Q.M.,Peng X.F.,and Liu G.C.,1994,Inheritance of aroma in rice,Sichuan Nongye Daxue Xuebao(Journal of Sichuan Agricultural University),12(3):392-395(任光俊,李青茂,彭兴富,刘光春,1994,水稻香味的遗传,四川农业大学学报,12(3):392-395)
Sakthivel K.,Sundaram R.M.,Shobha Rani N.,Balachandran S.M.,and Neeraja C.N.,2009,Genetic and molecular basis of fragrance in rice,Biotechnol.Adv.,27(4):468-473
Sansenya S.,Hua Y.,Chumanee S.,Phasai K.,and Sricheewin C.,2017,Effect of gamma irradiation on 2-acetyl-1-pyrrolinecontent,GABA content and volatile compounds of germinated rice(Thai upland rice),Plants,6(2):E18
Shao G.N.,Tang A.,Tang S.Q.,Luo J.,Jiao G.A.,Wu J.L.,and Hu P.S.,2011,A new deletion mutation of fragrant gene and the development of three molecular markers for fragrance in rice,Plant Breeding,130(2):172-176
Shi W.W.,Yang Y.,Chen S.H.,and Xu M.L.,2008,Discovery of a new fragrance allele and the development of functional markers for the breeding of fragrant rice varieties,Mol.Breeding,22(2):185-192
Shi Y.Q.,Zhao G.C.,Xu X.L.,and Li J.Y.,2014,Discovery of a new fragrance allele and development of functional markers for identifying diverse fragrant genotypes in rice,Mol.Breeding,33(3):701-708
Wang F.,Li J.H.,Liu W.G.,Liao Y.L.,Zhu M.S.,Liu Z.R.,Huang H.J.,and Huang D.J.,2008,Development of a functional marker for fragrance gene in rice,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),22(4):347-352(王丰,李金华,柳武革,廖亦龙,朱满山,刘振荣,黄慧君,黄德娟,2008,一种水稻香味基因功能标记的开发,中国水稻科学,22(4):347-352)
Widjaja R.,Craske J.D.,and Wooton M.,1996,Comparative studies on volatile components of non-fragrant and fragrant rices,Sci.Food Agric.,70(2):151-161
Zeng Y.,Xia X.Z.,Nong B.X.,Zhang Z.Q.,Yang X.H.,Deng G.F.,Liu K.Q.,Huang Z.,and Li D.T.,2017,Identification of aroma and aroma genotype in local specific fragrant rice varieties from Guangxi,Nanfang Nongye Xuebao(Journal of Southern Agriculture),48(9):1548-1553(曾宇,夏秀忠,农保选,张宗琼,杨行海,邓国富,刘开强,黄志,李丹婷,2017,广西特色香稻地方品种香味及其香味基因型的鉴定,南方农业学报,48(9):1548-1553)
Bindusree G.,Natarajan P.,Kalva S.,and Madasamy P.,2017,Whole genome sequencing of Oryza sativa L.cv.Seeragasamba identifies a new fragrance allele in rice,PLoS One,12(11):e0188920
Bourgis F.,Guyot R.,Gherbi H.,Tailliez E.,Amabile I.,Salse J.,Lorieux M.,Delseny M.,and Ghesquière A.,2008,Characterization of the major fragance gene from an aromatic japonica rice and analysis of its diversity in Asian cultivated rice,Theor.Appl.Genet.,117(3):353-368
Bradbury L.M.T.,Fitzgerald T.L.,Henry R.J.,Jin Q.S.,and Waters D.L.E.,2005,The gene for fragrance in rice,Plant Biotechnol.J.,3(3):363-370
Fitzgerald M.A.,Hamilton N.R.S.,Calingacion M.N.,Verhoeven H.A.,and Butardo V.M.,2008,Is there a second fragrance gene in rice?Plant Biotechnol.J.,6(4):416-423
Jiang D.,Lu X.Y.,Wang Y.C.,Fan Y.L.,Zhai W.X.,Luo Y.H.,and Xia Z.H.,2015,Sequence analysis of mutation site of Badh2 in 27 varieties of fragrant rice,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(2):276-280(姜达,卢小勇,王延春,范玉龙,翟文学,罗越华,夏志辉,2015,27种香稻品种badh2突变位点序列的分析,分子植物育种,13(2):276-280)
Kovach M.J.,Calingacionb M.N.,Fitzgeraldb M.A.,and McCoucha S.R.,2009,The origin and evolution of fragrance in rice(Oryza sativa L.),Proc.Natl.Acad.Sci.USA,106(34):14444-14449
Myint K.M.,Courtois B.,Risterucci A.M.,Frouin J.,Soe K.,Thet K.M.,Vanavichit A.,and Glaszmann J.C.,2012,Specific patterns of genetic diversity among aromatic rice varieties in Myanmar,Rice,5(1):1-13
Ootsuka K.,Takahashi I.,Tanaka K.,Itani T.,Tabuchi H.,Yoshihashi T.,Tonouchi A.,and Ishikawa R.,2014,Genetic polymorphisms in Japanese fragrant landraces and novel fragrant alleledomesticated in northern Japan,Breeding Science,64(2):115-124
Ren G.J.,Li Q.M.,Peng X.F.,and Liu G.C.,1994,Inheritance of aroma in rice,Sichuan Nongye Daxue Xuebao(Journal of Sichuan Agricultural University),12(3):392-395(任光俊,李青茂,彭兴富,刘光春,1994,水稻香味的遗传,四川农业大学学报,12(3):392-395)
Sakthivel K.,Sundaram R.M.,Shobha Rani N.,Balachandran S.M.,and Neeraja C.N.,2009,Genetic and molecular basis of fragrance in rice,Biotechnol.Adv.,27(4):468-473
Sansenya S.,Hua Y.,Chumanee S.,Phasai K.,and Sricheewin C.,2017,Effect of gamma irradiation on 2-acetyl-1-pyrrolinecontent,GABA content and volatile compounds of germinated rice(Thai upland rice),Plants,6(2):E18
Shao G.N.,Tang A.,Tang S.Q.,Luo J.,Jiao G.A.,Wu J.L.,and Hu P.S.,2011,A new deletion mutation of fragrant gene and the development of three molecular markers for fragrance in rice,Plant Breeding,130(2):172-176
Shi W.W.,Yang Y.,Chen S.H.,and Xu M.L.,2008,Discovery of a new fragrance allele and the development of functional markers for the breeding of fragrant rice varieties,Mol.Breeding,22(2):185-192
Shi Y.Q.,Zhao G.C.,Xu X.L.,and Li J.Y.,2014,Discovery of a new fragrance allele and development of functional markers for identifying diverse fragrant genotypes in rice,Mol.Breeding,33(3):701-708
Wang F.,Li J.H.,Liu W.G.,Liao Y.L.,Zhu M.S.,Liu Z.R.,Huang H.J.,and Huang D.J.,2008,Development of a functional marker for fragrance gene in rice,Zhongguo Shuidao Kexue(Chinese Journal of Rice Science),22(4):347-352(王丰,李金华,柳武革,廖亦龙,朱满山,刘振荣,黄慧君,黄德娟,2008,一种水稻香味基因功能标记的开发,中国水稻科学,22(4):347-352)
Widjaja R.,Craske J.D.,and Wooton M.,1996,Comparative studies on volatile components of non-fragrant and fragrant rices,Sci.Food Agric.,70(2):151-161
Zeng Y.,Xia X.Z.,Nong B.X.,Zhang Z.Q.,Yang X.H.,Deng G.F.,Liu K.Q.,Huang Z.,and Li D.T.,2017,Identification of aroma and aroma genotype in local specific fragrant rice varieties from Guangxi,Nanfang Nongye Xuebao(Journal of Southern Agriculture),48(9):1548-1553(曾宇,夏秀忠,农保选,张宗琼,杨行海,邓国富,刘开强,黄志,李丹婷,2017,广西特色香稻地方品种香味及其香味基因型的鉴定,南方农业学报,48(9):1548-1553)