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

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Abstract
Proteinase inhibitor I was 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 distributed between the vacuoles and cytoplasm of induced leaves. Low vacuole to leaf cell ratios of 3 cytoplasmic markers, triosephosphate isomerase, catalase and chlorophyll, indicated that the isolated vacuoles were relatively free of intact protoplasts and cell debris.