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Study on glutelin genes of storage proteins in rice.

水稻贮藏蛋白谷蛋白基因的研究.

基本信息

批准号：
04454041
负责人：
SATOH Hikaru
金额：
$ 4.1万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454041/
关键词：
Rice Improvement of grain properties Genetic resources Storage protein Glutelin Gene analysis SDS-PAGE IEF グリテリン SOS-PAGE 突然変異体

项目摘要

Two major subunits of rice glutelin, the acidic (alpha) and basic (beta) subunits consist of various polypeptides in their apparent molecular mass and pI.We found a wide variation among the rice atrains in the apparent molecular mass and pI by SDSPAGE and IEF,especially between Japonica and Indica types. The glutelin of "Kinmaze", Japonica type, and "IR36", Indica type, were analyzed exactly. In SDSPAGE analysis, the glutelin was separated into four alpha subnit bands, alpha-1, alpha-2, alpha-3 and alpha-4, and four beta subunit bands, beta-1, beta-2, beta-3 and beta-4.The alpha-3 of "Kinmaze" moved faster than that of "IR36". In IEF analysis, alpha and beta subunits had at least 12 and 9 bands, respectively. Several polymorphic hands were detected in both subunits. The relation between the apparent molecular mass and pI of the polypeptides were analyzed by 2-dimensional electrophoresis. Each SDS-PAGE band consists of some polypeptides showing pI heterogeneity, while some polypeptides … More of same pI showed different molecular weight. The polymorphic bands in IEF were detected as each single spot, and belong to alpha-2 and beta-2. The genetic analysis of the polymorphic bands were done by IEF using "Kinmaze", "IR24" and RI (recombinant inbred) lines. "IR24" shows almost the same pattern with "IR36", but has another band in alpha subunit. RI lines were made from a cross seed between "Asominori", showing the same pattern with "Kinmaze", and "IR24". The bands of reciprocal F1 hybrid seeds between "Kinmaze"and "IR24"showed the gene dosage effect. In F2 analysis, segregation of the bands, phenotypes in staining intensity, fitted to a ratio of 1 : 1 : 1 : 1 (or 3 : 1), indicating these bands were controlled by incomplete dominant genes. Recombinants were found among the bands, indicating each band was controlled by each incomplete gene. The linkage relationship of the genes controlling each polymorphic bands were found in both genes controlling alpha-2 polypeptides and alpha-3 and beta-2 polypeptides. Then chromosomal location of these genes was studied by means of RI lines. The genes controlling alpha-2 polypeptides were shown to be linked with RFLP markers located on the chromosome 1, while The genes controlling alpha-3 and beta-2 polypeptides were shown to be linked with RFLP markers located on the chromosome 2. Less

谷蛋白的两个主要亚基，酸性亚基（α）和碱性亚基（β）在其表观分子质量和pI上由不同的多肽组成。通过SDSPAGE和IEF分析发现，水稻系间表观分子质量和表观分子质量存在较大差异，籼粳品种间差异较大。对粳稻型“Kinmaze”和籼稻型“IR36”的谷蛋白进行了精确分析。在SDSPAGE分析中，将谷蛋白分离为α -1、α -2、α -3和α -4四个α亚基带，以及β -1、β -2、β -3和β -4四个β亚基带。“Kinmaze”的alpha-3移动速度比“IR36”快。在IEF分析中，α亚基和β亚基分别至少有12个和9个波段。在两个亚基中均检测到多态手。通过二维电泳分析了多肽的表观分子质量与pI之间的关系。每条SDS-PAGE带由一些具有pI异质性的多肽组成，而相同pI的多肽更多表现出不同的分子量。在IEF中检测到的多态性条带为单个点，分别属于α -2和β -2。以“Kinmaze”、“IR24”和RI（重组自交系）为材料，利用IEF对多态性条带进行遗传分析。“IR24”与“IR36”表现出几乎相同的模式，但在α亚基上有另一个条带。以“Asominori”与“Kinmaze”和“IR24”的杂交种子为杂交材料，获得了与“Kinmaze”相同的杂交株系。“Kinmaze”与“IR24”互易F1杂交种子条带表现出基因剂量效应。在F2分析中，条带的分离，染色强度的表型，符合1:1:1:1（或3:1）的比例，表明这些条带由不完全显性基因控制。在条带之间发现了重组，表明每个条带由每个不完整基因控制。在α -2多肽和α -3和β -2多肽的基因中均发现了控制各多态性带的基因连锁关系。然后用RI系研究了这些基因的染色体定位。结果表明，控制α -2多肽的基因与位于1号染色体上的RFLP标记相连，而控制α -3和β -2多肽的基因与位于2号染色体上的RFLP标记相连。少