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Plant and Cell Physiology, 1997, Vol. 38, No. 9 1069-1074
© 1997

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

Yoshiko Minami1, Hiroyasu Takao1, Takeo Kanafuji1, Kazuo Miura1, Maki Kondo2,3, Ikuko Hara-Nishimura2,3, Mikio Nishimura2,3 and Hiroshi Matsubara1

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

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 against the rßglucosidase clearly showed that the rß-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. rßGlucosidase 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-rßglucosidase antibodies. The rß-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 rß-glucosidase is tissue specific in its expression in the leaves of the indigo plant.

(Received April 14, 1997; Accepted July 10, 1997)
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