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The rates of CO/sub 2/ evolution from Douglas-fir (Pseudotsuga menziesii )Mirb.) Franco) needle litter, following application of divalent metal (Hg, Cd, Pb, Ni, Zn, and Cu) chlorides at rates of 10, 100, and 1000 ..mu..g/g and Ca chloride at 7, 68, and 683 ..mu..g/g, were monitored at 2- to 3-day intervals for 4 weeks; extractable enzymatic activities were also measured on additional samples at 1 day, 2, and 4 weeks following treatment. All metals except Pb inhibited respiration when applied at the highest level. Cellulase activity was depressed by treatments of 1000 ..mu..g/g of either Hg or Cd after 4 weeks but was not affected initially (1 day after). After 4 weeks, xylanase activity was also depressed by the 1000-..mu..g/g Hg treatment. Apparently these enzymatic activities declined due to decreased enzyme synthesis associated with inhibited microbial growth rather than by direct enzymatic inhibition. Although addition of Hg or Cu chlorides directly to litter extracts caused marked inhibition of most enzymatic activities, equivalent levels, when applied to the needle litter, produced no inhibitions in extracts after 1 day, except for invertase activity. Invertase activity was depressed at all intervals and ..beta..-glucosidase activity was elevated at all intervals by Hg at themore » 1000-..mu..g/g level. The level of extractable ninhydrin-reactive compounds was elevated by this Hg treatment at all intervals similar to the effect of fumigation of soil or litter. Regardless of treatment, cellulase and xylanase activities were correlated (r = 0.70, n = 192).« less

Assays for measuring the activities of cellulase, xylanase, mannase, amylase, ..beta..-glucosidase, invertase, and protease employing buffered suspensions of ground coniferous and deciduous leaf litter exhibited zero-order kinetics. Only a small percentage of the whole-litter activities of invertase, ..beta..-glucosidase, and protease were extractable into 0.05M potassium acetate, pH 5.0; however, extractable activities of cellulase and xylanase represented from 39 to 174% of the whole-litter activities indicating their soluble exocellar nature. Extractable protease and amylase activities were best correlated with the average daily rates of CO/sub 2/ evolution in a group of 90 leaf litter samples equally representing 18 coniferous species. Enzymatic activities were readily detectable in extracts of all samples but classification of the samples by species provided little differentiation in the distribution of either enzymatic activities or rates of CO/sub 2/ evolution. Mannase, cellulase, and xylanase activities were well-correlated with each other in all samples. Assays attempting to measure a pool of readily-metabolizable substances in litter by extractable reducing substances, ninhydrin-positive substances, glucose, and phenolics failed to show correlation coefficients >0.41 with rates of CO/sub 2/ evolution. Addition of D-(+)-catechin to litter extracts, up to levels equivalent to those observed in the group of samples, did not inhibit any carbohydrasemore » thus suggesting the lack of inhibition of litter-decomposing enzymes by the concentrations of phenolics present in these coniferous leaf litters.« less

Douglas fir (Pseudotsuga menziesii (Mirb.) Franco) needle-litter was given treatments of varying biocidal severity including air-drying, oven-drying, fumigation with methyl bromide plus chloropicrin. autoclaving, and a control. The litter was inoculated with a small amount of fresh litter homogenate and monitored for CO2 evolution rate and extractable enzymatic activity during a 77-day incubation. Enzymatic activities assayed included cellulase. xylanase, mannase, amylase. invertase, β-glucosidase, protease, polyphenoloxidase and peroxidase. The concentrations of extractable ninhydrin-reactive compounds, glucose, and reducing substances were also determined. Air-drying produced a small, initial stimulation in respiration but had no effect on any enzymatic activity throughout the 77-day incubation except for an initial decrease in polyphenoloxidase and peroxidase activities. Oven-drying produced a substantial loss of all enzymatic activities initially but all carbohydrase and protease activities recovered to greater than control levels by 7 days; after a 1-day lag, oven-drying produced a pronounced stimulation in the rate of CO2 evolution. Autoclaving produced a 2-day lag in a less pronounced (compared to oven-drying) respiratory stimulation: initially all enzymatic activities were destroyed and only protease and β-glucosidase activities recovered to control levels. Fumigation produced a respiratory stimulation after a 9-day lag (probably due to residual chloropicrin) which coincided with a pronounced increase in cellulase. xylanase, mannase and amylase activities: initially, no activities except polyphenoloxidase and peroxidase were adversely affected by fumigation. During the entire incubation, the activities of cellulase. xylanase. mannase. amylase and invertase were highly correlated with each other regardless of treatment. After 77 days, the rates of CO2 evolution of all samples were correlated with the level of amylase and cellulase activity (r = 0.86 and 0.83, respectively). This observation points to the value of enzymatic assays for following the processes which release substrate for microbial assimilation from plant litter. The close association of polysaccharidase and protease activities with the flush of CO2 evolution following biocidal treatment, particularly oven-drying, indicates that the decomposition of carbohydrates and proteins released from microbial biomass or plant litter is a mechanism for this often-observed respiratory stimulation.