Abstract
We have isolated nodule specific genes from Astragalus sinicus which forms indeterminate-type nodules. One of the clones, AsNODf32, encodes a cysteine protease that strongly expresses in a senescent zone of the nodules. It is important to compare AsNODf32 with homologues from other legumes that form determinate-type nodule for understanding the functions. Therefore, we identified GNf037h07 from EST database of model leguminous plant Lotus japonicus. In this study, we constructed a plasmid which expresses GNf037h07:GFP fusion protein constitutively to examine the subcellular localization of GNf037h07. The fusion protein expressed in cultured cells of tobacco BY-2 localized in cytoplasm of rapidly growing cells and in the vacuole of senescent cells. These findings suggest that GNf037h07 works in the vacuoles of mature nodules. We are now studying leader sequence of the protease that determines growth-phase-dependent subcellular localization of the protease using GFP fusion proteins.
