Lipids and lipolytic enzymes in the trunkwood ofRobinia pseudoacacia L. during heartwood formation

Abstract

The radial profile of lipase and phospholipase activities was determined in the trunkwood ofRobinia pseudoacacia L. The trees were felled in November at the time of heartwood formation and alterations in the enzymatic activities were investigated across the sapwood and heartwood. Methods employed include gaschromatographic, colorimetric and enzymatic assays. On a dry weight basis, the hydrolysis of the artificial substrate pnitrophenylpalmitate shows a maximum activity in growth ring 4; however, the assay has proved not to be specific for lipase. In contrast, lipase analyses (triacylglycerol acylhydrolase; E.C. 3.1.1.3) with an authentic substrate show activity peaks in growth rings 1 and 4. With protein as a reference the highest activity is found in growth ring 5. A similar tendency is observed for phospholipase A1 (E.C. 3.1.1.32) and phospholipase A2 (E.C. 3.1.1.4). Phospholipase C (E.C. 3.1.4.3) activity decreases towards the sapwood-heartwood boundary; negligible traces of activity are detected in the heartwood, whereas, based on the protein content, growth ring 4 yields maximal activity. Phospholipase D (3.1.4.4) exhibits the same radial pattern with regard to protein content as a reference. On a dry weight basis there is a significant increase within the sapwood area, while in the heartwood the activity drastically decreases. The enzyme activities are discussed in relation to degradative processes within the plasma membranes and the hydrolysis of reserve lipids during heartwood formation.