7份贵州禾香味鉴定及OsBADH2基因CDS差异分析
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摘要
水稻OsBADH2基因功能丧失是稻米产生香味的内因。本研究对7份贵州禾和2份对照品种的香味性状进行物化鉴定,发现榕禾与2个对照品种为香稻,其余6份贵州禾都为非香稻。为解析贵州禾产生香味与OsBADH2基因的内在关系,通过TA克隆、测序分析和序列拼接,本研究获得9份材料OsBADH2基因CDS的序列信息,采用Megalign软件对其进行序列比对分析。结果表明:4份贵州禾(白香禾2号、半坡禾、黎平禾1号和黎平禾2号)的OsBADH2基因CDS无突变;榕禾和2个对照品种的CDS差异都为第7外显子的8 bp缺失和3 SNP;还发现2种新的CDS差异(白香禾1号为1 bp缺失和1 SNP,便禾为2 bp缺失和2 SNP)。这些结果为贵州禾香味品种保护及育种改良提供理论基础。
Dysfunction of OsBADH2 gene in rice is the internal cause of rice to have fragrance. This study applied physicochemical methods to identify the fragrance trait of 7 Guizhou HE(Oryza sativa) and 2 control varieties. It was found that Rong He and 2 control varieties were fragrant rice, and the other 6 varieties were all non-fragrant rice.To analysis the internal relationship between fragrant production in Guizhou HE and OsBADH2 gene, the study obtained the CDS sequences of OsBADH2 gene in 9 tested materials, and conducted alignment analysis of these sequences by Megalign software through TA-cloning, sequencing analysis and sequence assembly. The results showed that the OsBADH2 gene CDS of 4 Guizhou HE(Baixianghe 2, Banpo He, Liping He 1 and Liping He 2) were normal. The CDS differences of Rong He and 2 control varieties were both 8 bp deletion and 3 SNP in exon 7, and2 new CDA differences were found which Baixianghe 1 was 1 bp deletion and 1 SNP, and Bian He was 2 bp deletion and 2 SNP. These results might provide the theoretical basis for varieties protection and breeding improvement of Guizhou HE(Oryza sativa).
Dysfunction of OsBADH2 gene in rice is the internal cause of rice to have fragrance. This study applied physicochemical methods to identify the fragrance trait of 7 Guizhou HE(Oryza sativa) and 2 control varieties. It was found that Rong He and 2 control varieties were fragrant rice, and the other 6 varieties were all non-fragrant rice.To analysis the internal relationship between fragrant production in Guizhou HE and OsBADH2 gene, the study obtained the CDS sequences of OsBADH2 gene in 9 tested materials, and conducted alignment analysis of these sequences by Megalign software through TA-cloning, sequencing analysis and sequence assembly. The results showed that the OsBADH2 gene CDS of 4 Guizhou HE(Baixianghe 2, Banpo He, Liping He 1 and Liping He 2) were normal. The CDS differences of Rong He and 2 control varieties were both 8 bp deletion and 3 SNP in exon 7, and2 new CDA differences were found which Baixianghe 1 was 1 bp deletion and 1 SNP, and Bian He was 2 bp deletion and 2 SNP. These results might provide the theoretical basis for varieties protection and breeding improvement of Guizhou HE(Oryza sativa).
引文
Bradbury L.M.T.,Fitzgerald T.L.,Henry R J,Liu X.G.,and Xu M.L.,2005,The gene for fragrance in rice,Plant Biotechnol.J.,3(3):363-370
Chen S.H.,Yang Y.,Shi W.W.,Ji Q.,He F.,Zhang Z.D.,Cheng Z.K.,Liu X.G.,and Xu M.D.,2008,Badh2,Encoding betaine aldehyde dehydrogenase,inhibits the biosynthesis of2-acetyl-1-pyrroline,a major component in rice fragrance,Plant Cell,20(7):1850-1861
Dhulappanavar C.V.,1976,Inheritance of scent in rice,Euphytica,25(1):659-662
Grimm C.C.,Bergman C.,Delgado J.T.,and Bryant R.,2001,Screening for 2-acetyl-1-pyrroline in the headspace of rice using SPME/GC-MS,Agric.Food.Chem.,49(1):245-249
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)
Ruan R.C.,Chen H.C.,Zhang Z.X.,Yang Y.S.,Jin T.Y.,You J.M.,and Zhu Y.Q.,2001,Progress and prospects of research and utilization on genetic diversity of rice germplasm in Guizhou,Zhiuwu Yichuan Ziyuan Kexue(Journal of Plant Genetic Resources),2(2):56-60(阮仁超,陈惠查,张再兴,杨玉顺,金桃叶,游俊梅,朱玉琴,2001,贵州稻种资源遗传多样性研究现状与展望,植物遗传资源科学,2(2):56-60)
Sakthivel K.,Sundaram R.M.,Shobha R.N.,Balachandran S.M.,and Neeraja C.N.,2009,Genetic and molecular basis of fragrance in rice,Biotechnol Adv.,27:468-473
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):185-19
Sood B.C.,and Siddiq E.A.,1978,A rapid technique for scent determination in rice,Indian Journal of Genetics&Plant Breeding,38(2):268-275
Xu X.L.,Zhao G.Q.,and Li J.Y.,2011,Development of molecular markers used to identify two types of fragrant rice and analysis of mutation sites of BADH2 gene in 24 varieties of fragrant rice,Zhiwu Fenlei Yu Ziyuan Xuebao(Plant Diversity and Resources),33(6):667-673(徐小龙,赵国超,李建粤,2011,24种香稻品种甜菜碱醛脱氢酶2基因突变位点的分析及分子标记开发,植物分类与资源学报,33(6):667-673)
Yajima L.,Yanai T.,Nakamura M.,Sakakibara H.,and Hayashi K.,1979,Volatile flavor components of cooked Kaorimai(scented rice,O.sativa japonica),Agric.Biol.Chem.,43:2425-2429
Chen S.H.,Yang Y.,Shi W.W.,Ji Q.,He F.,Zhang Z.D.,Cheng Z.K.,Liu X.G.,and Xu M.D.,2008,Badh2,Encoding betaine aldehyde dehydrogenase,inhibits the biosynthesis of2-acetyl-1-pyrroline,a major component in rice fragrance,Plant Cell,20(7):1850-1861
Dhulappanavar C.V.,1976,Inheritance of scent in rice,Euphytica,25(1):659-662
Grimm C.C.,Bergman C.,Delgado J.T.,and Bryant R.,2001,Screening for 2-acetyl-1-pyrroline in the headspace of rice using SPME/GC-MS,Agric.Food.Chem.,49(1):245-249
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)
Ruan R.C.,Chen H.C.,Zhang Z.X.,Yang Y.S.,Jin T.Y.,You J.M.,and Zhu Y.Q.,2001,Progress and prospects of research and utilization on genetic diversity of rice germplasm in Guizhou,Zhiuwu Yichuan Ziyuan Kexue(Journal of Plant Genetic Resources),2(2):56-60(阮仁超,陈惠查,张再兴,杨玉顺,金桃叶,游俊梅,朱玉琴,2001,贵州稻种资源遗传多样性研究现状与展望,植物遗传资源科学,2(2):56-60)
Sakthivel K.,Sundaram R.M.,Shobha R.N.,Balachandran S.M.,and Neeraja C.N.,2009,Genetic and molecular basis of fragrance in rice,Biotechnol Adv.,27:468-473
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):185-19
Sood B.C.,and Siddiq E.A.,1978,A rapid technique for scent determination in rice,Indian Journal of Genetics&Plant Breeding,38(2):268-275
Xu X.L.,Zhao G.Q.,and Li J.Y.,2011,Development of molecular markers used to identify two types of fragrant rice and analysis of mutation sites of BADH2 gene in 24 varieties of fragrant rice,Zhiwu Fenlei Yu Ziyuan Xuebao(Plant Diversity and Resources),33(6):667-673(徐小龙,赵国超,李建粤,2011,24种香稻品种甜菜碱醛脱氢酶2基因突变位点的分析及分子标记开发,植物分类与资源学报,33(6):667-673)
Yajima L.,Yanai T.,Nakamura M.,Sakakibara H.,and Hayashi K.,1979,Volatile flavor components of cooked Kaorimai(scented rice,O.sativa japonica),Agric.Biol.Chem.,43:2425-2429