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

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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.