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Immunological Identification of Proteinase Inhibitors I and II in Isolated Tomato Leaf Vacuoles ¹

^a Department of Agricultural Chemistry and Program in Biochemistry and Biophysics, Washington State University, Pullman, Washington 99164

Proteinase inhibitor I has been identified and quantified in isolated vacuoles from tomato (Lycopersicon esculentum) leaves induced to accumulate inhibitors either by wounding or by supplying excised leaves with the wound hormone, proteinase inhibitor-inducing factor. Proteinase inhibitor II was also identified in the vacuoles but not quantified. Control vacuoles were prepared from unwounded plants that did not contain inhibitors. Vacuole to leaf cell ratios of inhibitors, chlorophyll, and several vacuolar and cytoplasmic enzymes were determined. The inhibitors were found almost entirely in the vacuoles. Acid phosphatase was located in control leaf vacuoles, but was found in both vacuoles and cytoplasm in induced leaves. Carboxypeptidase, induced by wounding, was found distributed between the vacuoles and cytoplasm of induced leaves. Low vacuole to leaf cell ratios of three cytoplasmic markers, triosephosphate isomerase, catalase, and chlorophyll, indicated that the isolated vacuoles were relatively free of intact protoplasts and cell debris.

¹ Supported in part by National Science Foundation Grants GB-37972 and PCM-75-23629, and United States Department of Agricultural Cooperative States Research Service Grant 316-15-60. Scientific Paper No. 4742, Project 1791, College of Agriculture Research Center, Washington State University.

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