Spatial and functional correlation between diamine-oxidase and peroxidase activities and their dependence upon de-etiolation and wounding in chick-pea stems

Abstract

The activities of diamine oxidase (DAO, EC 1.4.3.6) and peroxidase (POD, EC 1.11.1.7) were determined along the stems of light-grown Cicer arietinum L. (chick-pea) seedlings. Enzyme activities were evaluated in the soluble, lightly bound (salt extraction) and tightly bound (Driselase digestion) wall fractions, and in residual fractions obtained from the different internodes. Apparent tissue distributions of both enzymes and lignin depositions were visualised by means of histochemical and immunohistochemical techniques. A close relationship was found between DAO and POD activities in the soluble and wall fractions along the stem. The biochemical activities of both enzymes decreased from the base to the apex of the stem in parallel with the distribution pattern of lignifying tissues in this organ. A similar activity gradient was found for each enzyme along the epidermis of the whole organ. Moreover, deetiolation elicited a rise in the activities of both enzymes in this tissue. Wounding chick-pea stems induced parallel increases in DAO and POD activities in the soluble and wall fractions. In-situ histochemical detection of both enzymes demonstrated the parallel occurrence of the DAO/POD system and lignosuberised depositions in the cell walls adjacent to the wound site. The patterns of POD isoforms resulting from the wound-healing process were determined by means of starch-gel electrophoresis. In addition to changes in relative intensity of enzyme bands in soluble and wall fractions, a new POD isoform, possibly related to the wounding response, appeared in the soluble fraction. This isoform was shown to be lightly bound to cell walls as it could be detected in the extracellular fluids obtained from wound-healed seedlings. On the basis of the above-mentioned results, a strict spatial and functional correlation can be inferred between DAO and POD in chick-pea, and probably in other Leguminosae species, in accordance with previous evidence indicating an integrated role for these enzymes in the apoplast.