Proteolytic activity in relation to seasonal cambial growth and xylogenesis in Pinus banksiana

Proteolytic activity in the cambial zone and developing xylem of Pinus banksiana Lamb. was investigated over an annual cycle of growth and dormancy. Highest proteolytic activity was associated with the most active period of primary-wall radial expansion of cambial derivatives, in early spring, before protoplasmic autolysis was initiated in developing earlywood. Three pH maxima of proteolytic activity, near pH 3.0, 6.5 and 9.5, were observed at that time. In general, activities measured at pH values below 7.0 were greater than those determined above pH 7.0 at all stages in the annual cycle, in both cambial zone and developing xylem, although elevated activity at alkaline pH was also observed during springtime growth. Polyvinylpolypyrollidone (PVP) treatment markedly enhanced pH 7.5 but not pH 4.0 proteolytic activity in the cambial zone, but not in developing xylem, indicating the presence of PVP-binding proteinase regulators in the cambium. By fractionation and effector studies total proteolysis was determined to comprise interactions between serine, cystine, aspartate and metallo-proteases having MWs, by gel chromatography, between 10 and 100 kDa. The observations point to a complex regulatory mechanism controlling the presence and catalytic rates of the distinct types of proteases in the cambial region throughout an annual cycle of growth and dormancy.