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Plant and Cell Physiology, 1994, Vol. 35, No. 5 821-827
© 1994

Characterization of Accumulation of Tobacco PR-5 Proteins by IEF-Immunoblot Analysis

Hisashi Koiwa, Fumihiko Sato1 and Yasuyuki Yamada

Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University Kyoto, 606 Japan

1To whom all correspondence should be addressed.

Accumulation of group 5 pathogenesis-related (PR) proteins in tobacco was characterized by immunological analysis combined with isoelectric focusing. This method clearly distinguished 3 subclasses of PR-5 proteins; basic (osmotin), neutral (osmotin-like protein: OLP) and acidic (PRS). A high accumulation of PR-5 proteins was detected only in root tissues in which neutral osmotin-like protein was mainly accumulated, in addition to a small amount of osmotin. Immunohistochemical analysis revealed that OLP accumulated in the cortex of the root. Cultured tobacco cells accumulated large amounts of neutral PR-5 proteins (OLP) in cells and the acidic form (PR-S) in the medium. Adaptation to salt stress was associated with a higher accumulation of basic PR-5 (osmotin) and less neutral PR-5.

In an analysis of the effect of biotic and abiotic stimuli on the synthesis of PR-5 proteins in leaf tissues, ethylene was found to induce a high accumulation of basic and neutral PR-5 proteins. Tobacco mosaic virus (TMV) infection induced accumulation of all major PR-5 proteins, but induction of OLP was less than that of the other isoforms. Systemic induction of PR-5 proteins in upper non-infected leaves was not observed. Salicylate induced only a small accumulation of PRS. These results indicate that each PR-5 protein has an independent regulatory pathway for its gene expression.

(Received December 7, 1993; Accepted May 11, 1994)
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