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

我们对细胞器的机制和细胞器之间的运输感兴趣。为了探讨拟南芥子叶等营养器官表皮细胞中特异性存在的内质网体的形成机制，对突变体进行了分析。超微结构分析和免疫电镜分析结果表明，A2突变株ER体的形成受阻，粗面内质网合成的GFP-HDEL不能从内质网腔排出。结果，GFP-HDEL在内质网腔内异常积累。在A28和B23突变株中，GFP-HDEL不停留在内质网腔内，而是从内质网排出。结果，GFP-HDEL被异常包装在形成的不同于ER体的卵泡结构中。推测可能有三个或三个以上的基因参与了ER体的正常形成和功能。通过对A28株系致病基因的初步定位，确定候选基因位于第三染色体上的YUP 4G 12 RE标记附近。通过对两个异常积累黑芥子酶的syntaxin（Vam 3）突变体的分析，发现黑芥子酶在液泡中大量积累，并有ER体样结构参与积累过程。此外，我们还研究了快速冷冻金属接触装置（HIF-4K）能否用于植物组织细胞的冷冻固定，并证实了加压冷冻装置（HPM-010）的成功率和质量均优于HIF-4K装置。

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.