Journal Article

rβ-Glucosidase in the Indigo Plant: Intracellular Localization and Tissue Specific Expression in Leaves

Yoshiko Minami, Hiroyasu Takao, Takeo Kanafuji, Kazuo Miura, Maki Kondo, Ikuko Hara-Nishimura, Mikio Nishimura and Hiroshi Matsubara

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 38, issue 9, pages 1069-1074
Published in print January 1997 | ISSN: 0032-0781
Published online January 1997 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/oxfordjournals.pcp.a029273
rβ-Glucosidase in the Indigo Plant: Intracellular Localization and Tissue Specific Expression in Leaves

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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rβ-Glucosidase of indigo plant (Polygonum tinctorium) has a high substrate specificity for indican (indoxyl rβ-D-gIu-coside). To examine the localization of this rβ-glucosidase, we fractionated the cells of the leaves and analysed them im-munocytochemically. Immunoelectron micrographs with specific antibodies against the rβglucosidase clearly showed that the rβ-glucosidase was localized in the stroma of the chloroplasts in mesophyll cells, but not in the thylakoid membrane. Chloroplasts were isolated from the crude ho-mogenate of the fresh leaves by Percoll density gradient centrifugation and then subjected to suborganellar frac-tionation. rβGlucosidase activity was specifically detected in the stromal fraction, but not in the thylakoid membrane. This was also supported by the result of an immunoblot of the fractions with anti-rβglucosidase antibodies. The rβ-gIu-cosidase was immunocytochemically localized in the chloroplasts of mesophyll cells, but not in any chloroplasts in marginal cells of the vascular bundle or epidermal cells; ribulose 1,5-bisphosphate carboxylase (Rubisco), a typical stromal protein, was observed in all chloroplasts in these cells. These results suggest that rβ-glucosidase is tissue specific in its expression in the leaves of the indigo plant.

Keywords: Chloroplasts; jff-Glucosidase (EC 3.2.1.21); Indican; Indigo; Polygonum tinctorium

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Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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