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Genetic mechanism controlling the transportation and accumulation of glutelin into protein body in rice endosperm

控制水稻胚乳中谷蛋白向蛋白体转运和积累的遗传机制

基本信息

批准号：
12138206
负责人：
SATOH Hikaru
金额：
$ 24.51万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2003
项目状态：
已结题

项目摘要

To elucidate the mechanism of intracellular transportation and accumulation of rice seed storage protein, 130 Iines of 57H mutant, which are characterized by the increased amount of proglutelin with 57 kD, were produced and novel 4 genes for 57H trait were identified. Based on genetic and histochemical analyses of the mutants, the function of each gene was discussed.-Mutation for the sorting of storage proteins within endoplasmic reticulum (ER), Type1 mutation The analysis of esp2 mutation deleted PDI demonstrated that PDI was indispensable for the sorting of proglutelin precursor and prolamin within ER Iumen. glup4 and glup6 mutations also were classified into Type1 mutation, showing that at least 3 genes participate in the sorting of both proteins within ER lumen.-Mutation for transportation from ER to protein storage vacuole (PSV), Type2 mutation In Type2 mutation, proglutelin precursor and prolamin were accumulated into separated particles within ER derived PB. Glup1, glup2, Glup5, glup7, Glup8 mutations were classified into Type2 mutation, showing that at least 5 genes participate in the transportation of proglutelin from ER lumen to PSV.-Mutation for proteolysis within PSV. Type3 mutation The analysis of glup3 mutation demonstrated that VPE was indispensable for proteolytical cleavage of proglutelin precursor within PSV. glup3 mutation accumulated the progiutelin and the cleaved form of glutelin in PSV together, showing that at least 2 VPEs participate in proteolysis of proglutelin precursor.These results demonstrated that the pathway of transportation and accumulation of glutelin in seed was divided into steps and that the genes regulating each step were 1) Targeting of glutelin mRNA to cis-ER, 2) Sorting of proglutelin precursor within ER lumen, Esp2 (PDI), Glup4, Glup6, 3) Transportation of proglutelin precursor into PSV, glupl, Glup2, glup5, Glup7, glup8, 4) Proteolysis of proglutelin precursor within PSV, Glup3 (VPE),

为了阐明水稻种子贮藏蛋白的细胞内转运和积累机制，我们培育了130株57H突变体，其特征是原谷蛋白含量增加，57kd，并鉴定了4个与57H性状相关的新基因。根据突变体的遗传和组织化学分析，讨论了每个基因的功能。-内质网（ER）储存蛋白分选突变，1型突变esp2突变缺失PDI的分析表明，PDI对于内质网（ER）内原谷蛋白前体和蛋白的分选是必不可少的。glup4和glup6突变也被归类为1型突变，表明至少有3个基因参与内质网管腔内这两种蛋白的分选。-从内质网转运到蛋白质储存液泡（PSV）的突变，2型突变在2型突变中，原谷朊蛋白前体和原蛋白在内质网衍生的PB中积聚成分离的颗粒。Glup1、glup2、Glup5、glup7、Glup8突变被归类为2型突变，表明至少有5个基因参与了原谷蛋白从ER管腔向PSV的转运。- PSV蛋白水解突变。对glup3突变的分析表明，VPE是PSV中原谷蛋白前体蛋白水解裂解所必需的。glup3突变将原谷蛋白和裂解形式的谷蛋白聚集在PSV中，表明至少有2个vpe参与了原谷蛋白前体的蛋白水解。这些结果表明，谷氨酸在种子中的转运和积累途径分为几个步骤，每个步骤的调控基因分别是：1)将谷氨酸mRNA靶向顺式内质网，2)内质网腔内的原谷氨酸前体Esp2 (PDI), Glup4, Glup6, 3)原谷氨酸前体转运到PSV， glupl, Glup2, glup5, Glup7, glup8, 4)原谷氨酸前体在PSV， Glup3 (VPE)，