Immunogold Localization of the Citrus Exocortis Viroid-Induced Pathogenesis-Related Proteinase P69 in Tomato Leaves

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Abstract
Citrus exocortis viroid induces in tomato plants (Lycopersicon esculentum) synthesis and accumulation of a pathogenesis-related protein (P69) previously reported to a proteinase (Vera P, Conejero V [1988] Plant Physiol 87: 58-63). By immunogold/transmission electron microscopy, we have studied the distribution of this protein in thin sections of parenchymatous leaf tissue. The enzyme was present intra- and extracellularly. The intracellular location was limited to the vacuole and was always associated with engulfed cell material. When extracellularly located, the enzyme was associated with a dispersed, electron-dense material in the intercellular spaces. This latter location was confirmed after analysis of intercellular washing fluids obtained by vacuum infiltration of leaves. These observations provide new data for the understanding of viroid pathogenesis and the biological role of the pathogenesis-related proteinase P69.