r{beta}-Glucosidase in the Indigo Plant: Intracellular Localization and Tissue Specific Expression in Leaves |
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Authors: | Minami, Yoshiko Takao, Hiroyasu Kanafuji, Takeo Miura, Kazuo Kondo, Maki Hara-Nishimura, Ikuko Nishimura, Mikio Matsubara, Hiroshi |
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Affiliation: | 1 Department of Biochemistry, Okayama University of Science 1-1 Ridai-cho, Okayama, 700 Japan 2 Department of Cell Biology, National Institute for Basic Biology Okazaki, 444 Japan 3 Department of Molecular Biomechanics, School of Life Science, The Graduate University for Advanced Studies Okazaki, 444 Japan |
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Abstract: | rß-Glucosidase of indigo plant (Polygonum tinctorium)has a high substrate specificity for indican (indoxyl rß-D-gIu-coside).To examine the localization of this rß-glucosidase,we fractionated the cells of the leaves and analysed them im-munocytochemically.Immunoelectron micrographs with specific antibodies againstthe rßglucosidase clearly showed that the rß-glucosidasewas localized in the stroma of the chloroplasts in mesophyllcells, but not in the thylakoid membrane. Chloroplasts wereisolated from the crude ho-mogenate of the fresh leaves by Percolldensity gradient centrifugation and then subjected to suborganellarfrac-tionation. rßGlucosidase activity was specificallydetected in the stromal fraction, but not in the thylakoid membrane.This was also supported by the result of an immunoblot of thefractions with anti-rßglucosidase antibodies. Therß-gIu-cosidase was immunocytochemically localizedin the chloroplasts of mesophyll cells, but not in any chloroplastsin marginal cells of the vascular bundle or epidermal cells;ribulose 1,5-bisphosphate carboxylase (Rubisco), a typical stromalprotein, was observed in all chloroplasts in these cells. Theseresults suggest that rß-glucosidase is tissue specificin its expression in the leaves of the indigo plant. (Received April 14, 1997; Accepted July 10, 1997) |
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