77. 重水で処理したシロイヌナズナ種子のプロテオミクス

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Germination commences when the dry seeds take up water. This Imbibition triggers various vital reactions, such as gibberellin biosynthesis, mobilization of stored endosperm reserves, and so on. To get new information about the various reactions involved in seed germination, we examined the effect of deuterium oxide (D_2O), which extremely delays germination, on proteome of Arabidopsis seed. Two-dimensional (2-D) gel electrophoresis was applied to find the difference between seeds imbibed H_2O and D_2O. As a result, degradation products of storage protein (CRA1, CRB) were decreased by D_2O treatment. We checked transcriptional levels of the enzymes (β VPE, δ VPE, carboxy peptidase, Asp protease) involved in the processing of the storage proteins by RT-PCR. It was revealed that transcripts levels of Ser protease and Cys protease were decreased by D_2O treatment. In addition, germination test with Ser protease inhibitors (AEBSF, aprotinin, PMSF) and Cys protease inhibitors (E-64d, antipain, leupeptin) were performed. A certain type of Ser protease inhibitors (AEBSF, PMSF) strongly repressed seed germination. This consists with the results of previous reports on other plants. Furthermore, 2-D gel electrophoresis was performed on the proteome of seeds treated with AEBSF. The same protein spots were decreased as well as D_2O treated seeds. On the other hand, Ser protease activity in D_2O was almost same as that in H_2O. These results suggest that Ser protease plays an important role in degradation of the storage proteins during seed germination and the effect of D_2O is caused by decreasing transcriptional levels of the Ser protease.
植物化学調節学会の論文
2004-10-13

77. 重水で処理したシロイヌナズナ種子のプロテオミクス

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