三个水稻突变体的遗传分析及单侧卷叶基因url1（t）的定位
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摘要
突变体是功能基因组学研究的重要材料,许多的突变体已用于基因的定位和克隆。本文鉴定了三个水稻突变体,斑点叶突变体M14、卷叶突变体B157和B369,分析了三个突变体的遗传特征,并初步定位了单侧卷叶基因urll(t)(unilateral rolled leaf 1)。主要结论如下:
1、从已构建的三个突变体的F_2群体中,观察各F_2群体中目标性状的分离情况,探明三个突变基因的显隐性关系和控制突变性状的基因数目。结果显示:斑点叶突变体M14和卷叶突变体B157是由一对等位基因控制的隐性遗传突变体,它们的分离比例符合孟德尔的一对等位基因的3∶1分离比例;卷叶突变体B369的F_2群体中分离出双亲类型的个体和中间类型个体,分离比例为1∶2∶1,推定为一对等位基因控制的不完全显性突变体。
2、以东洋超级稻×B157的F_2群体中的58株卷叶单株为定位群体,将单侧卷叶基因urll(t)初步定位在水稻第1染色体的30cM位置附近处,目前未见在该位点报道相关的卷叶基因,因此推断urll(t)是个新的卷叶基因。通过分子标记的连锁分析,找到了与urll(t)紧密连锁的标记RI02526和RM272,两标记与urll(t)的遗传距离为2.0cM和0.6cM。进一步的insilico分析显示,分子标记RI02526和RM272分别位于BAC克隆AP003200和AP003052,两标记间的物理距离为692.9kb,包含78个假定基因。
3、在TIGR基因组数据库中,查阅第1染色体上的目标区段内78个假定基因。发现在78个假定基因中有两个RNA依赖性的RNA聚合酶假定基因和三个与植物细胞分裂和生长有关的假定基因,但没有发现miRNA基因,也没有发现与叶片近轴化/远轴化发育相关的编码HD-ZIPⅢ转录因子的基因以及AGO蛋白的同源基因。两个编码RNA聚合酶的假定基因在TIGR的登陆号为Gene:11971.t00888和Gene:11971.t00889,两基因都位于克隆AP003200。三个与植物细胞分裂和生长有关的假定基因是:Gene:11971.t00886位于克隆AP003200,编码细胞分裂素脱氢酶的前体蛋白;Gene:11971.t00917位于克隆AP003197,编码与细胞分裂的相关蛋白;Gene:11971.t00935位于克隆AP003105,编码BES1/BZR1的同源蛋白,BES1/BZR1是植物生长素调控的关键转录因子。这五个基因可能与B157的卷叶发生有关。
Mutants were one of the most important materials for function genome research.Many of them had been used for gene mapping and gene cloning. Three rice mutants,which were spot leaf mutant M14、rolled leaf mutants B157 and B369,were identified by classical genetical analysis in this paper.Also,we had primarily mapped url1(t)(unilateral rolled leaf 1)to chromosome 1.The main results were here:
1、Six F_2 populations were developed from three mutants.In order to confirming whether the three mutants were controlled by recessive gene or dominant gene and controlled by a single gene or not,we investigated the segregation ratio of character in every population.It was suggest the mutant phenotypes of M14 and B157 were controlled by a single recessive gene,while B369 was controlled by a semi-dominant gene.
2、A mapping population including 58 mutant phenotype plants was derived from crossing between Super Rice in Toxyox(ssp.indica)and B157.With this mapping population,a rolled leaf gene url1(t)from B157 was primarily mapped to the site of 30cM on Chromosome 1.So far,It was not report there was a rolled leaf gene at this site,so we concluded that url1(t)was a new rolled leaf gene in rice.Two molecular markers RI02526 and RM272 closely linked to url1(t)were obtained on two sides,the genetic distance between them and url1(t) was 2.0cM and 0.6cM respectively.RI02526 and RM272 were located on BAC clones AP003200 and AP003052,a 692.9kb region containing url1(t)gene was defined with the two markers,including 78 putative genes.
3、Red all putative genes in the 692.9kb region in TIGR database annotating rice genome.We didn't find miRNA gene and the homologous gene coding HD-ZIPⅢand AGO proteins related to leaf-adaxial development,but we found five putative genes which might be associated with the rolled leaf gene url1(t). The five putative genes were gene:11971.t00888,gene:11971.t00889, gene:11971.t00886,gene:11971.t00917,gene:11971.t00935.gene:11971.t00888 and gene:11971.t00889 were located on BAC clones AP003200,coded RNA-dependent RNA polymerase family protein;Gene:11971.t00886 was located on BAC clones AP003200,coded cytokinin dehydrogenase 1 precursor; Gene:11971.t00917 was located on BAC clones AP003197,coded cell division related protein;Gene:11971.t00935 was located on BAC clones AP003105, coded BES1/BZR1 homolog protein 4,BES1/BZR1 was the key transcription factor in regulation of auxin.
1、从已构建的三个突变体的F_2群体中,观察各F_2群体中目标性状的分离情况,探明三个突变基因的显隐性关系和控制突变性状的基因数目。结果显示:斑点叶突变体M14和卷叶突变体B157是由一对等位基因控制的隐性遗传突变体,它们的分离比例符合孟德尔的一对等位基因的3∶1分离比例;卷叶突变体B369的F_2群体中分离出双亲类型的个体和中间类型个体,分离比例为1∶2∶1,推定为一对等位基因控制的不完全显性突变体。
2、以东洋超级稻×B157的F_2群体中的58株卷叶单株为定位群体,将单侧卷叶基因urll(t)初步定位在水稻第1染色体的30cM位置附近处,目前未见在该位点报道相关的卷叶基因,因此推断urll(t)是个新的卷叶基因。通过分子标记的连锁分析,找到了与urll(t)紧密连锁的标记RI02526和RM272,两标记与urll(t)的遗传距离为2.0cM和0.6cM。进一步的insilico分析显示,分子标记RI02526和RM272分别位于BAC克隆AP003200和AP003052,两标记间的物理距离为692.9kb,包含78个假定基因。
3、在TIGR基因组数据库中,查阅第1染色体上的目标区段内78个假定基因。发现在78个假定基因中有两个RNA依赖性的RNA聚合酶假定基因和三个与植物细胞分裂和生长有关的假定基因,但没有发现miRNA基因,也没有发现与叶片近轴化/远轴化发育相关的编码HD-ZIPⅢ转录因子的基因以及AGO蛋白的同源基因。两个编码RNA聚合酶的假定基因在TIGR的登陆号为Gene:11971.t00888和Gene:11971.t00889,两基因都位于克隆AP003200。三个与植物细胞分裂和生长有关的假定基因是:Gene:11971.t00886位于克隆AP003200,编码细胞分裂素脱氢酶的前体蛋白;Gene:11971.t00917位于克隆AP003197,编码与细胞分裂的相关蛋白;Gene:11971.t00935位于克隆AP003105,编码BES1/BZR1的同源蛋白,BES1/BZR1是植物生长素调控的关键转录因子。这五个基因可能与B157的卷叶发生有关。
Mutants were one of the most important materials for function genome research.Many of them had been used for gene mapping and gene cloning. Three rice mutants,which were spot leaf mutant M14、rolled leaf mutants B157 and B369,were identified by classical genetical analysis in this paper.Also,we had primarily mapped url1(t)(unilateral rolled leaf 1)to chromosome 1.The main results were here:
1、Six F_2 populations were developed from three mutants.In order to confirming whether the three mutants were controlled by recessive gene or dominant gene and controlled by a single gene or not,we investigated the segregation ratio of character in every population.It was suggest the mutant phenotypes of M14 and B157 were controlled by a single recessive gene,while B369 was controlled by a semi-dominant gene.
2、A mapping population including 58 mutant phenotype plants was derived from crossing between Super Rice in Toxyox(ssp.indica)and B157.With this mapping population,a rolled leaf gene url1(t)from B157 was primarily mapped to the site of 30cM on Chromosome 1.So far,It was not report there was a rolled leaf gene at this site,so we concluded that url1(t)was a new rolled leaf gene in rice.Two molecular markers RI02526 and RM272 closely linked to url1(t)were obtained on two sides,the genetic distance between them and url1(t) was 2.0cM and 0.6cM respectively.RI02526 and RM272 were located on BAC clones AP003200 and AP003052,a 692.9kb region containing url1(t)gene was defined with the two markers,including 78 putative genes.
3、Red all putative genes in the 692.9kb region in TIGR database annotating rice genome.We didn't find miRNA gene and the homologous gene coding HD-ZIPⅢand AGO proteins related to leaf-adaxial development,but we found five putative genes which might be associated with the rolled leaf gene url1(t). The five putative genes were gene:11971.t00888,gene:11971.t00889, gene:11971.t00886,gene:11971.t00917,gene:11971.t00935.gene:11971.t00888 and gene:11971.t00889 were located on BAC clones AP003200,coded RNA-dependent RNA polymerase family protein;Gene:11971.t00886 was located on BAC clones AP003200,coded cytokinin dehydrogenase 1 precursor; Gene:11971.t00917 was located on BAC clones AP003197,coded cell division related protein;Gene:11971.t00935 was located on BAC clones AP003105, coded BES1/BZR1 homolog protein 4,BES1/BZR1 was the key transcription factor in regulation of auxin.
引文
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