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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

水稻谷蛋白的两个主要亚基，酸性亚基(α亚基)和碱性亚基(β亚基)由不同表观分子质量和等电点的多肽组成。SDSPAGE和IEF分析发现，不同水稻品种的表观分子质量和等电点存在较大差异，特别是在粳型和籼型之间。较准确地分析了“金迷宫”和“IR36”两个品种的谷蛋白含量。在SDSPAGE分析中，谷蛋白被分成4个α亚基条带，即α-1、α-2、α-3和α-4，以及4个β亚基条带，即β-1、β-2、β-3和β-4。在IEF分析中，α和β亚基分别至少有12条和9条带。在这两个亚基中都检测到了多个多态的手。用双向电泳法分析了多肽的表观分子质量与等电点的关系。每条SDSPAGE条带均由一些显示等电点异质性的多肽组成，而一些多肽…等电点相同的多表现为不同的相对分子质量。在IEF中，每个点都检测到了多态条带，属于α-2和β-2。用IEF对“Kinmaze”、“IR24”和RI(重组自交系)的多态条带进行遗传分析。“IR24”表现出与“IR36”几乎相同的模式，但在阿尔法亚基中有另一条带。RI系是由“Asominori”与“Kinmaze”和“IR24”的杂交种子配制而成的。“金迷子”与“IR24”正反交种子的条带表现出一定的基因剂量效应。在F2分析中，显带的分离和染色强度的表型符合1：1：1：1(或3：1)的比例，表明这些带是由不完全显性基因控制的。在这些条带中发现了重组子，表明每个条带都是由不完全基因控制的。在控制α-2多肽和控制α-3和β-2多肽的基因中都发现了控制每个多态条带的基因的连锁关系。然后利用RI系对这些基因进行了染色体定位研究。控制α-2多肽的基因与位于第1染色体的RFLP标记连锁，而控制α-3和β-2多肽的基因与位于第2染色体的RFLP标记连锁。