Inheritance of alleles for glutelin α-2 subunit genes in rice and identification of their corresponding cDNA clone
Inheritance of alleles for glutelin α-2 subunit genes in rice and identification of their corresponding cDNA clone
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DOI：
10.1007/s00122-002-1046-6
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发表时间：
2002-12-01
影响因子：
5.4
通讯作者：
Takaiwa, F
中科院分区：
文献类型：
--
作者：
Qu, LQ;Wei, XL;Takaiwa, F
水稻谷蛋白由酸性（α）和碱性（β）亚基组成，其进一步分为三种多肽组分，分别称为α-1、α-2和α-3亚基组分以及β-1、β-2和β-3亚基组分。用SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）方法，对约6,800个N-甲基-N-亚硝基脲（MNU）处理水稻受精卵的潜在突变株系进行筛选，获得了9个谷蛋白α-2亚基组分（α-2L）含量降低的突变株系。将突变体分为三种类型：增加的α-1亚基（α-1H/α-2L）、减少的β-2亚基（β-2L/α-2L）和增加的α-3亚基（α-3H/α-2L），分别以EM 278、CM 1707和EM 659为代表。等电聚焦（IEF）分析显示，所有突变体都具有极少量的具有6.71 pI值的多肽，而具有6.50 pI值或具有6.90 pI值的多肽分别在α-1H/α-2L突变体或α-3H/α-2L突变体中显著增加。β-2L/α-2L突变体的碱性多肽量减少，pI值为8.74。遗传分析表明，这三种突变体均受一对不完全显性基因控制，均为等位基因。该基因被暂时命名为glu 4，它被发现位于1号染色体上，与eg和sp16基因连锁。双向电泳分析显示，glu 4编码的多肽pI 6.71/α-2和pI 8.74/β-2。氨基酸序列分析表明，突变的酸性多肽是GluA亚家族基因的产物。北方和RT-PCR分析表明，glu 4对应于GluA-1基因。
Rice glutelins consist of acidic (alpha) and basic (beta) subunits which are further separated into three polypeptide components assigned as alpha-1, alpha-2, and alpha-3 subunit components and beta-1, beta-2 and beta-3 subunit components. Nine rice mutant lines with a decreased amount of the glutelin alpha-2 subunit component (alpha-2L) were obtained by screening about 6,800 potential mutant lines derived from the fertilized egg treatment with N-methyl-N-nitrosourea (MNU) using the SDS-PAGE method. The mutants were classified into three types of the increased alpha-1 subunit (alpha-1H/alpha-2L), the decreased beta-2 subunit (beta-2L/alpha-2L) and the increased alpha-3 subunit (alpha-3H/alpha-2L) represented by EM278, CM1707 and EM659, respectively. Iso-electric focus (IEF) analysis revealed that all of the mutants had an extremely low amount of a polypeptide with a 6.71 pI value, whereas a polypeptide with either a 6.50 pI value or with a 6.90 pI value increased significantly in alpha-1H/alpha-2L mutants or in alpha-3H/alpha-2L mutants, respectively. The beta-2L/alpha-2L mutants had a decreased amount of a basic polypeptide with a 8.74 pI value. Genetic analysis revealed that the three types of mutants were controlled by a single incomplete dominant gene respectively, and the three are alleles. The gene was temporarily named glu4, which was found to be located on chromosome 1 linked with the eg and sp16 genes. Two-dimensional electrophoresis analysis revealed that the glu4 encoded polypeptides of pI 6.71/alpha-2 and pI 8.74/beta-2. Amino acid sequence analysis suggested that the mutated acidic polypeptide was the product of a GluA subfamily gene. Northern and RT-PCR analyses revealed that glu4 corresponded to the GluA-1 gene.