水稻簇生穗基因OsCl-6的精细定位与候选基因分析
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摘要
水稻(Oryza sativa)穗部性状的改良对水稻产量提高和品种提升具有重要意义。水稻簇生穗的研究有助于了解水稻的穗发育以及相应基因的调控方式。本研究通过构建籼稻(O.sativa ssp.indica)簇生穗材料内江P164和单粒稻9311的正交和反交群体,确定该性状为单基因不完全显性遗传。通过混合群体分离分析法和隐性群体分析法将该基因定位于第6染色体长臂标记ID441和ID020之间,区间内预测有23个基因。推测LOC_Os06g39650为候选基因,并对候选基因进行克隆,测序表明该基因(命名为:水稻簇生穗基因(O.sativa clustered spikelets gene-6,OsCl-6))CDS的第1 556的碱基发生了突变,由A突变为G,碱基的改变造成氨基酸序列发生突变,第519位的氨基酸由天冬酰胺(N)突变为丝氨酸(S),推测从而形成簇生穗性状,进一步对6个非簇生粳稻品种和7个非簇生籼稻品种进行测序分析,结果表明,OsCl-6基因可能是簇生穗基因短花梗(shortened pedicels)sped1-D的等位基因。本研究为阐明水稻穗簇生形成机理以及选育穗大、着粒密度高的水稻新品种提供了基因资源。
The improvement of panicle traits in rice(Oryza sativa) is of great significance to the promotion of rice yield and varieties. The study of characteristics of clustered spikelet helps to understand spikelet development and its corresponding gene expression and regulation. In this study, phenotype analysis of reciprocal crosses population between Neijiang P164 and 9 311 indicated that the phenotype of clustered spikelet in Neijiang P164 was controlled by a single incomplete dominant gene. Combining bulked segregate analysis(BSA) and recessive class analysis(RCA), the gene was mapped between markers ID441 and ID020 in the 6 th chromosome of rice, which contains 23 annotated genes in this region. LOC_Os06 g39650 was selected as candidate gene. Cloning and sequencing of the candidate gene indicated that there was a single base substituted from A to G at the 1 556 thof coding region, resulting in a substitution from asparagine(N) to serine(S) at amino acid site of 519 th. Furthermore, the candidate gene(named as(O. sativa clustered spikelets gene-6, OsCl-6)) of 13 non-clustered varieties including 6 indica and 7 japonica rice varieties were sequenced.The result indicated that OsCl-6 gene might be the allele of clustered spikelet gene shortened pedicels1-D(sped1-D). This study provide a germplasm resource for mechanism analysis of clustered spikelet and breeding new rice varieties with large panicle and high grain density.
The improvement of panicle traits in rice(Oryza sativa) is of great significance to the promotion of rice yield and varieties. The study of characteristics of clustered spikelet helps to understand spikelet development and its corresponding gene expression and regulation. In this study, phenotype analysis of reciprocal crosses population between Neijiang P164 and 9 311 indicated that the phenotype of clustered spikelet in Neijiang P164 was controlled by a single incomplete dominant gene. Combining bulked segregate analysis(BSA) and recessive class analysis(RCA), the gene was mapped between markers ID441 and ID020 in the 6 th chromosome of rice, which contains 23 annotated genes in this region. LOC_Os06 g39650 was selected as candidate gene. Cloning and sequencing of the candidate gene indicated that there was a single base substituted from A to G at the 1 556 thof coding region, resulting in a substitution from asparagine(N) to serine(S) at amino acid site of 519 th. Furthermore, the candidate gene(named as(O. sativa clustered spikelets gene-6, OsCl-6)) of 13 non-clustered varieties including 6 indica and 7 japonica rice varieties were sequenced.The result indicated that OsCl-6 gene might be the allele of clustered spikelet gene shortened pedicels1-D(sped1-D). This study provide a germplasm resource for mechanism analysis of clustered spikelet and breeding new rice varieties with large panicle and high grain density.
引文
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周军杰,李兴星,郑剑,等.2016.水稻粒簇生材料Cgr320的鉴定及遗传偏分离分析[J].植物遗传资源学报,17(5):942-950.(Zhou J J,Li X X,Zheng J,et al.2016.Observation and genetic segregation distortion analysis on clustered spikelets in rice resource Cgr320[J].Journal of Plant Genetic Resources,17(5):942-950.)
周鹏,邓其明,涂诗航,等.2012.水稻穗部簇生突变体(Cldz)的遗传分析与初步定位[J].福建农业学报,27(5):481-484.(Zhou P,Deng Q M,Tu S H,et al.2012.Genetic analysis and gene mapping of the clustered spikelets mutant(Cl-dz)of rice[J].Fujian Journal of Agricultural Sciences,27(5):481-484.)
Guo M,Yang Y H,Liu M,et al.2014.Clustered spikelets 4,encoding a putative cytochrome P450 protein CYP724B1,is essential for rice panicle development[J].Chinese Science Bulletin,59(31):4050-4059.
Jiang G H,Xiang Y H,Zhao J Y,et al.2014.Regulation of inflorescence branch development in rice through a novel pathway involving the pentatricopeptide repeat protein sped1-D[J].Genetics,197:1395-1407.
Jodon N E.1957.Inheritance of some of the more striking characters in rice[J].Heredity,48(4):73-101.
Nagao S,Takahashi M.1963.Trial construction of twelve linkage groups in Japanese rice(Genetical studies on rice Plant,XXVII)[J].Journal of the Faculty of Agriculture,53(1):72-130.
Zheng L Y,Zhu X D,Qian Q,et al.2003.Morphology and mapping analysis of rice(Oryza sativa L.)clustered spikelets(Cl)mutant[J].Chinese Science Bulletin,48(6):559-562.
陈红旗,刘刚,朱旭东,等.2002.水稻簇生穗突变体的鉴定及遗传[J].南京农业大学学报,25(3):116-118.(Chen H Q,Liu G,Zhu X D,et al.2002.Observation and genetically analysis on character of clustered spikelets in rice[J].Journal of Nanjing Agricultural University,25(3):116-118.)
孟浪.2011.水稻簇生穗突变体cll(t)的基因定位及候选基因筛选[D].硕士学位论文,福建农林大学,导师:王锋,pp.2.(Meng L.2011.Gene mapping and candidategene screening of a clustered spikelet mutant cll(t)in rice[D].Thesis for M.S.,Fujian Agricultural and Forestry University,Supervisor:Wang F,pp.2.)
田翠,张涛,蒋开锋,等.2010.水稻小穗簇生突变体的遗传分析及其基因的初步定位[J].分子植物育种,8(1):29-34.(Tian C,Zhang T,Jiang K F,et al.2010.Genetic analysis and preliminary gene mapping of rice clustered spikelets mutant[J].Molecular Plant Breeding,8(1):29-34.)
肖巧珍.2007.水稻簇生穗簇生基因的等位性鉴定和遗传分析[D].硕士学位论文,广西大学,导师:刘丕庆,pp.34.(Xiao Q Z.2007.Allelism test and inheritance analysis of rice clustered spikelets mutant gene[D].Thesis for M.S.,Guangxi University,Supervisor:Liu P Q,pp.34.)
曾秀红.2009.水稻簇生穗基因cl-4的精细定位[D].硕士学位论文,扬州大学,导师:严长杰,pp.1.(Zeng X H.2009.Fine mapping of a clustered spikelet gene cl-4 in Rice[D].Thesis for M.S.,Yangzhou University,Supervisor:Yan C J,pp.1.)
张现伟,李经勇,郑家奎,等.2013.复粒稻种质资源及其遗传育种利用[J].植物学报,48(4):438-446.(Zhang XW,Li J Y,Zheng J K,et al.2013.Germplasm resources and genetic breeding utilization of multiple-grain rice[J].Chinese Bulletin of Bontany,48(4):438-446.)
张艳丽,向珣朝,姚嫣萍,等.2010.水稻顶小穗双粒突变体TS的颖花发育和簇生穗发生特征[J].中国种业,12:44-46.(Zhang Y L,Xiang X C,Yao Y P,et al.2010.Spikelet development and cluster panicle characteristics of a rice top spikelet double mutant TS[J].China Seeds,12:44-46.)
张毅,李云峰,谢戎,等.2006.水稻小穗簇生性近等基因系的构建及其近等性评价[J].作物学报,32(3):397-401.(Zhang Y,Li Y F,Xie R,et al.2006.Development of NILs with Cl-gene of rice restorer and evaluation on the Near-isogenic level[J].Acta Agronomica Sinica,32(3):397-401.)
周军杰,李兴星,郑剑,等.2016.水稻粒簇生材料Cgr320的鉴定及遗传偏分离分析[J].植物遗传资源学报,17(5):942-950.(Zhou J J,Li X X,Zheng J,et al.2016.Observation and genetic segregation distortion analysis on clustered spikelets in rice resource Cgr320[J].Journal of Plant Genetic Resources,17(5):942-950.)
周鹏,邓其明,涂诗航,等.2012.水稻穗部簇生突变体(Cldz)的遗传分析与初步定位[J].福建农业学报,27(5):481-484.(Zhou P,Deng Q M,Tu S H,et al.2012.Genetic analysis and gene mapping of the clustered spikelets mutant(Cl-dz)of rice[J].Fujian Journal of Agricultural Sciences,27(5):481-484.)
Guo M,Yang Y H,Liu M,et al.2014.Clustered spikelets 4,encoding a putative cytochrome P450 protein CYP724B1,is essential for rice panicle development[J].Chinese Science Bulletin,59(31):4050-4059.
Jiang G H,Xiang Y H,Zhao J Y,et al.2014.Regulation of inflorescence branch development in rice through a novel pathway involving the pentatricopeptide repeat protein sped1-D[J].Genetics,197:1395-1407.
Jodon N E.1957.Inheritance of some of the more striking characters in rice[J].Heredity,48(4):73-101.
Nagao S,Takahashi M.1963.Trial construction of twelve linkage groups in Japanese rice(Genetical studies on rice Plant,XXVII)[J].Journal of the Faculty of Agriculture,53(1):72-130.
Zheng L Y,Zhu X D,Qian Q,et al.2003.Morphology and mapping analysis of rice(Oryza sativa L.)clustered spikelets(Cl)mutant[J].Chinese Science Bulletin,48(6):559-562.