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Functional analysis of organelle in the cotyledonous cells of Arabidopsis thaliana under the ultrastructural research.

超微结构研究下拟南芥子叶细胞细胞器的功能分析。

基本信息

批准号：
15570048
负责人：
HAYASHI Yasuko
金额：
$ 2.37万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2006
项目状态：
已结题

项目摘要

We have interested in the mechanism of organelle and the transport between organelle. To examine the formation mechanism of ER body that existed in the peculiarity in epidermal cell of the nourishment organ such as cotyledon of Arabidopsis thaliana, the ER-body form mutant was analyzed. Our data from ultra-microscopic analysis and immune-electron microscopic analysis revealed that the formation of ER-body was obstructed in A2 mutant line, and GFP-HDEL synthesized by the rough endoplasmic reticulum was not discharged from the endoplasmic reticulum lumen. As a result, GFP-HDEL had accumulated abnormally in the endoplasmic reticulum lumen. In A28 and the B23 mutant lines, GFP-HDEL did not stay in the endoplasmic reticulum lumen and was discharged from endoplasmic reticulum. As a result, GFP-HDEL had being packed into the follicle structure formed differing from ER body abnormally. It was guessed that three genes or more took part from this for a normal formation and the function of ER body. From the rough mapping of the cause gene about A28 line, we identified the candidate gene existed in the vicinity of the YUP4G12RE marker on the third chromosome. From the analysis of two syntaxin (Vam3) mutants that abnormality accumulated myrosinases, we found that the myrosin cell had been accumulated voluminously in vacuole and ER-body-like structure took part in the accumulation process. It was suggested that ER-body carry out various functions by the difference of the tissues.In addition to these, we examined whether rapid frozen metal contact device (HIF-4K) was able to be used for the frozen fixation of the plant organization cell, and it was confirmed that both success rates and the qualities of pressurizing frozen device (HPM-010) were more effective than HIF-4K device.

我们对细胞器的机制和细胞器之间的运输很感兴趣。为了探讨存在于拟南芥子叶等营养器官表皮细胞中的ER小体的形成机制，对ER小体形态突变体进行了分析。超微结构分析和免疫电子显微镜分析结果表明，A2突变系ER小体的形成受阻，粗面内质网合成的GFP-HDEL没有从内质网腔中排出。结果GFP-HDEL在内质网腔内异常积聚。在A28和B23突变株中，GFP-HDEL不停留在内质网上，而是从内质网上排出。因此，GFP-HDEL被包裹在卵泡结构中，形成了不同于内质网小体的结构。推测有三个或更多的基因参与了内质网的正常形成和内质网的功能。通过对A28系的致病基因进行粗略定位，确定了位于第3染色体上YUP4G12RE标记附近的候选基因。通过对两个异常积累黑芥子酶的合成素(Vam3)突变体的分析，发现黑芥子细胞在液泡中大量积累，内质网体样结构参与了积累过程。此外，我们还考察了快速冷冻金属接触器(HIF-4K)能否用于植物组织细胞的冷冻固定，证实加压冷冻装置(HPM-010)的成功率和质量都优于HIF-4K装置。