Decomposition of [C]Lignocelluloses of Spartina alterniflora and a Comparison with Field Experiments

Decomposition of lignocelluloses from Spartina alterniflora in salt-marsh sediments was measured by using C-labeled compounds. Rates of decomposition were fastest in the first 4 days of incubation and declined later. Lignins labeled in side chains were mineralized slightly faster than uniformly labeled lignins; 12% of the [side chain-C]lignin-labeled lignocellulose was mineralized after 816 h of incubation, whereas only 8% of the [U-C]lignin-labeled lignocelluloses were degraded during this period. The carbohydrate moiety within the lignocellulose complex was degraded about four times faster than the lignin moiety; after 816 h of incubation, 29 to 37% of the carbohydrate moiety had been mineralized. Changes in concentration of lignin and cellulose in litter of S. alterniflora were followed over 2 years of decay. Cellulose disappeared from litter more rapidly than lignin; 50% of the initial content of cellulose was lost after 130 days, whereas lignin required 330 to 380 days for 50% loss. The slow loss of lignin compared with other litter components resulted in a progressive enrichment of litter in lignin content. The rates of mineralization of [C]lignocelluloses in marsh sediments were similar to the rates of lignocellulose decomposition in litter on the marsh.     

Mineralization of Detrital Lignocelluloses by Salt Marsh Sediment Microflora

Specifically radiolabeled ¹⁴C-(cellulose)-lignocellulose and ¹⁴C-(lignin)-lignocellulose were isolated from labeled cuttings of Spartina alterniflora (cordgrass) and Pinus elliottii (slash pine). These were used to estimate the rates of mineralization to CO₂ of lignocelluloses of estuarine and terrestrial origin in salt marsh estuarine sediments. The lignin moiety of pine lignocellulose was mineralized 10 to 14 times more slowly than that of Spartina lignocellulose, depending on the source of inoculum. Average values for percent mineralization after 835 h of incubation were 1.4 and 13.9%, respectively. For Spartina lignocellulose, mineralization of the cellulose moiety was three times faster than that of the lignin moiety. Average values for percent mineralization after 720 h of incubation were 32.1 and 10.6%, respectively. Lignocellulose and lignin contents of live pine and Spartina plants were analyzed and found to be 60.7 and 20.9%, respectively, for pine and 75.6 and 15.1%, respectively, for Spartina.     

Isolation of a Bacterium Capable of Degrading Peanut Hull Lignin

Thirty-seven bacterial strains capable of degrading peanut hull lignin were isolated by using four types of lignin preparations and hot-water-extracted peanut hulls. One of the isolates, tentatively identified as Arthrobacter sp., was capable of utilizing all four lignin preparations as well as extracted peanut hulls as a sole source of carbon. The bacterium was also capable of degrading specifically labeled [¹⁴C]lignin-labeled lignocellulose and [¹⁴C]cellulose-labeled lignocellulose from the cordgrass Spartina alterniflora and could also degrade [¹⁴C]Kraft lignin from slash pine. After 10 days of incubation with [¹⁴C]cellulose-labeled lignocellulose or [¹⁴C]lignin-labeled lignocellulose from S. alterniflora, the bacterium mineralized 6.5% of the polysaccharide component and 2.9% of the lignin component.     

Preparation, Characterization, and Microbial Degradation of Specifically Radiolabeled [14C]Lignocelluloses from Marine and Freshwater Macrophytes

Specifically radiolabeled [¹⁴C-lignin]lignocelluloses were prepared from the aquatic macrophytes Spartina alterniflora, Juncus roemerianus, Rhizophora mangle, and Carex walteriana by using [¹⁴C]phenylalanine, [¹⁴C]tyrosine, and [¹⁴C]cinnamic acid as precursors. Specifically radiolabeled [¹⁴C-polysaccharide]lignocelluloses were prepared by using [¹⁴C]glucose as precursor. The rates of microbial degradation varied among [¹⁴C-lignin]lignocelluloses labeled with different lignin precursors within the same plant species. To determine the causes of these differential rates, [¹⁴C-lignin]lignocelluloses were thoroughly characterized for the distribution of radioactivity in nonlignin contaminants and within the lignin macromolecule. In herbaceous plants, significant amounts (8 to 24%) of radioactivity from [¹⁴C]phenylalanine and [¹⁴C]tyrosine were found associated with protein, although very little (3%) radioactivity from [¹⁴C]cinnamic acid was associated with protein. Microbial degradation of radiolabeled protein resulted in overestimation of lignin degradation rates in lignocelluloses derived from herbaceous aquatic plants. Other differences in degradation rates among [¹⁴C-lignin]lignocelluloses from the same plant species were attributable to differences in the amount of label being associated with ester-linked subunits of peripheral lignin. After acid hydrolysis of [¹⁴C-polysaccharide]lignocelluloses, radioactivity was detected in several sugars, although most of the radioactivity was distributed between glucose and xylose. After 576 h of incubation with salt marsh sediments, 38% of the polysaccharide component and between 6 and 16% of the lignin component (depending on the precursor) of J. roemerianus lignocellulose was mineralized to ¹⁴CO₂; during the same incubation period, 30% of the polysaccharide component and between 12 and 18% of the lignin component of S. alterniflora lignocellulose was mineralized.     

Anaerobic Biodegradation of the Lignin and Polysaccharide Components of Lignocellulose and Synthetic Lignin by Sediment Microflora

Specifically radiolabeled [¹⁴C-lignin]lignocelluloses and [¹⁴C-polysaccharide]lignocelluloses were prepared from a variety of marine and freshwater wetland plants including a grass, a sedge, a rush, and a hardwood. These [¹⁴C]lignocellulose preparations and synthetic [¹⁴C]lignin were incubated anaerobically with anoxic sediments collected from a salt marsh, a freshwater marsh, and a mangrove swamp. During long-term incubations lasting up to 300 days, the lignin and polysaccharide components of the lignocelluloses were slowly degraded anaerobically to ¹⁴CO₂ and ¹⁴CH₄. Lignocelluloses derived from herbaceous plants were degraded more rapidly than lignocellulose derived from the hardwood. After 294 days, 16.9% of the lignin component and 30.0% of the polysaccharide component of lignocellulose derived from the grass used (Spartina alterniflora) were degraded to gaseous end products. In contrast, after 246 days, only 1.5% of the lignin component and 4.1% of the polysaccharide component of lignocellulose derived from the hardwood used (Rhizophora mangle) were degraded to gaseous end products. Synthetic [¹⁴C]lignin was degraded anaerobically faster than the lignin component of the hardwood lignocellulose; after 276 days, 3.7% of the synthetic lignin was degraded to gaseous end products. Contrary to previous reports, these results demonstrate that lignin and lignified plant tissues are biodegradable in the absence of oxygen. Although lignocelluloses are recalcitrant to anaerobic biodegradation, rates of degradation measured in aquatic sediments are significant and have important implications for the biospheric cycling of carbon from these abundant biopolymers.     

Relative Contributions of Bacteria and Fungi to Rates of Degradation of Lignocellulosic Detritus in Salt-Marsh Sediments

Specifically radiolabeled [¹⁴C-lignin]lignocellulose and [¹⁴C-polysaccharide]lignocellulose from the salt-marsh cordgrass Spartina alterniflora were incubated with an intact salt-marsh sediment microbial assemblage, with a mixed (size-fractionated) bacterial assemblage, and with each of three marine fungi, Buergenerula spartinae, Phaeosphaeria typharum, and Leptosphaeria obiones, isolated from decaying S. alterniflora. The bacterial assemblage alone mineralized the lignin and polysaccharide components of S. alterniflora lignocellulose at approximately the same rate as did intact salt-marsh sediment inocula. The polysaccharide component was mineralized twice as fast as the lignin component; after 23 days of incubation, ca. 10% of the lignin component and 20% of the polysaccharide component of S. alterniflora lignocellulose were mineralized. Relative to the total sediment and bacterial inocula, the three species of fungi mediated only very slow mineralization of the lignin and polysaccharide components of S. alterniflora lignocellulose. Experiments with uniformly ¹⁴C-labeled S. alterniflora material indicated that the three fungi and the bacterial assemblage were capable of degrading the non-lignocellulosic fraction of S. alterniflora material, but only the bacterial assemblage significantly degraded the lignocellulosic fraction. Our results suggest that bacteria are the predominant degraders of lignocellulosic detritus in salt-marsh sediments.     

Thermophilic Anaerobic Biodegradation of [14C]Lignin, [14C]Cellulose, and [14C]Lignocellulose Preparations

Thermophilic (55°C) anaerobic enrichment cultures were incubated with [¹⁴C-lignin]lignocellulose, [¹⁴C-polysaccharide]lignocellulose, and kraft [¹⁴C]lignin prepared from slash pine, Pinus elliottii, and ¹⁴C-labeled preparations of synthetic lignin and purified cellulose. Significant but low percentages (2 to 4%) of synthetic and natural pine lignin were recovered as labeled methane and carbon dioxide during 60-day incubations, whereas much greater percentages (13 to 23%) of kraft lignin were recovered as gaseous end products. Percentages of label recovered from lignin-labeled substrates as dissolved degradation products were approximately equal to percentages recovered as gaseous end products. High-pressure liquid chromatographic analyses of CuO oxidation products of sound and degraded pine lignin indicated that no substantial chemical modifications of the remaining lignin polymer, such as demethoxylation and dearomatization, occurred during biodegradation. The polysaccharide components of pine lignocellulose and purified cellulose were relatively rapidly mineralized to methane and carbon dioxide; 31 to 37% of the pine polysaccharides and 56 to 63% of the purified cellulose were recovered as labeled gaseous end products. An additional 10 to 20% of the polysaccharide substrates was recovered as dissolved degradation products. Overall, these results indicate that elevated temperatures can greatly enhance rates of anaerobic degradation of lignin and lignified substrates to methane and low-molecular-weight aromatic compounds.     

Preparation, characterization, and microbial degradation of specifically radiolabeled [C]lignocelluloses from marine and freshwater macrophytes

Specifically radiolabeled [C-lignin]lignocelluloses were prepared from the aquatic macrophytes Spartina alterniflora, Juncus roemerianus, Rhizophora mangle, and Carex walteriana by using [C]phenylalanine, [C]tyrosine, and [C]cinnamic acid as precursors. Specifically radiolabeled [C-polysaccharide]lignocelluloses were prepared by using [C]glucose as precursor. The rates of microbial degradation varied among [C-lignin]lignocelluloses labeled with different lignin precursors within the same plant species. To determine the causes of these differential rates, [C-lignin]lignocelluloses were thoroughly characterized for the distribution of radioactivity in nonlignin contaminants and within the lignin macromolecule. In herbaceous plants, significant amounts (8 to 24%) of radioactivity from [C]phenylalanine and [C]tyrosine were found associated with protein, although very little (3%) radioactivity from [C]cinnamic acid was associated with protein. Microbial degradation of radiolabeled protein resulted in overestimation of lignin degradation rates in lignocelluloses derived from herbaceous aquatic plants. Other differences in degradation rates among [C-lignin]lignocelluloses from the same plant species were attributable to differences in the amount of label being associated with ester-linked subunits of peripheral lignin. After acid hydrolysis of [C-polysaccharide]lignocelluloses, radioactivity was detected in several sugars, although most of the radioactivity was distributed between glucose and xylose. After 576 h of incubation with salt marsh sediments, 38% of the polysaccharide component and between 6 and 16% of the lignin component (depending on the precursor) of J. roemerianus lignocellulose was mineralized to CO(2); during the same incubation period, 30% of the polysaccharide component and between 12 and 18% of the lignin component of S. alterniflora lignocellulose was mineralized.     

Effects of Petroleum Hydrocarbons on Plant Litter Microbiota in an Arctic Lake

The effects of petroleum hydrocarbons on the microbial community associated with decomposing Carex leaf litter colonized in Toolik Lake, Alaska, were examined. Microbial metabolic activity, measured as the rate of acetate incorporation into lipid, did not vary significantly from controls over a 12-h period after exposure of colonized Carex litter to 3.0 ml of Prudhoe Bay crude oil, diesel fuel, or toluene per liter. ATP levels of the microbiota became elevated within 2 h after the exposure of the litter to diesel fuel or toluene, but returned to control levels within 4 to 8 h. ATP levels of samples exposed to Prudhoe Bay crude oil did not vary from control levels. Mineralization of specifically labeled ¹⁴C-[lignin]-lignocellulose and ¹⁴C-[cellulose]-lignocellulose by Toolik Lake sediments, after the addition of 2% (vol/vol) Prudhoe Bay crude oil, motor oil, diesel fuel, gasoline, n-hexane, or toluene, was examined after 21 days of incubation at 10°C. Diesel fuel, motor oil, gasoline, and toluene inhibited ¹⁴C-[lignin]-lignocellulose mineralization by 58, 67, 67, and 86%, respectively. Hexane-treated samples displayed an increase in the rate of ¹⁴C-[lignin]-lignocellulose mineralization of 33%. ¹⁴C-[cellulose]-lignocellulose mineralization was inhibited by the addition of motor oil or toluene by 27 and 64%, respectively, whereas diesel fuel-treated samples showed a 17% increase in mineralization rate. Mineralization of the labeled lignin component of lignocellulose appeared to be more sensitive to hydrocarbon perturbations than was the labeled cellulose component.     

Effects of pH on Lignin and Cellulose Degradation by Streptomyces viridosporus

Lignocellulose degradation by Streptomyces viridosporus results in the oxidative depolymerization of lignin and the production of a water-soluble lignin polymer, acid-precipitable polymeric lignin (APPL). The effects of the culture pH on lignin and cellulose metabolism and APPL production by S. viridosporus are reported. Dry, ground, hot-water-extracted corn (Zea mays) lignocellulose was autoclaved in 1-liter reagent bottles (5 g per bottle) and inoculated with 50-ml volumes of S. viridosporus cells suspended in buffers of specific pH (pH 6.0 to 9.2 at 0.4 pH unit intervals). Four replicates of inoculated cultures and of uninoculated controls at each pH were incubated as solid-state fermentations at 37°C. After 6 weeks of incubation the percent loss of lignocellulose, lignin, and carbohydrate and the amount of APPL produced were determined for each replicate. Optimal lignocellulose degradation, as shown by substrate weight loss, was observed in the pH range of 8.4 to 8.8. Only minor differences were seen in the Klason lignin, carbohydrate, protein, and ash contents of the APPLS produced by cultures at each pH. The effects of pH on the degradation of a spruce (Picea pungens) [¹⁴C-lignin]lignocellulose and a Douglas fir (Pseudotsuga menziesii) [¹⁴C-glucan]-lignocellulose were also determined at pH values between 6.5 and 9.5 (0.5 pH unit intervals). The incubations were carried out for 3 weeks at 37°C with bubbler-tube cultures. The percentage of initial ¹⁴C recovered as ¹⁴CO₂, ¹⁴C-labeled water-soluble products, and [¹⁴C]APPL was then determined. The mineralization of lignin and cellulose to CO₂ was optimal at pHs 6.5 and 7.0, respectively. However, the optimum for lignin and cellulose solubilization was pH 8.5, which correlated with the pH 8.5 optimum for APPL production. Overall, the data show that, whereas lignin mineralization is optimal at neutral to slightly acidic pHs, lignocellulose degradation with lignin solubilization and APPL production is promoted by alkaline pHs. These findings indicate that lignin-solubilizing actinomycetes may play an important role in the metabolism of lignin in neutral to alkaline soils in which ligninolytic fungi are not highly competitive.     

Microbial degradation of lignocellulose: the lignin component

A new procedure was developed for the study of lignin biodegradation by pure or mixed cultures of microorganisms. Natural lignocelluloses were prepared containing C in primarily their lignin components by feeding plants l-[U-C]phenylalanine through their cut stems. Lignin degradation was observed in numerous soils by monitoring evolution of CO(2) from [C]lignin-labeled oak (Quercus albus), maple (Acer rubrum), and cattail (Typha latifola). An organism (Thermonospora fusca ATCC 27730) that is known to degrade cellulose but not lignin was shown to grow on lignocellulose in the presence of [C]lignocelluloses without evolution of CO(2). A known lignin degrader (a white-rot fungus, Polyporus versicolor) was shown to readily evolve CO(2) from damp C-labeled cattail and C-labeled maple.     

Thermophilic anaerobic biodegradation of [C]lignin, [C]cellulose, and [C]lignocellulose preparations

Thermophilic (55 degrees C) anaerobic enrichment cultures were incubated with [C-lignin]lignocellulose, [C-polysaccharide]lignocellulose, and kraft [C]lignin prepared from slash pine, Pinus elliottii, and C-labeled preparations of synthetic lignin and purified cellulose. Significant but low percentages (2 to 4%) of synthetic and natural pine lignin were recovered as labeled methane and carbon dioxide during 60-day incubations, whereas much greater percentages (13 to 23%) of kraft lignin were recovered as gaseous end products. Percentages of label recovered from lignin-labeled substrates as dissolved degradation products were approximately equal to percentages recovered as gaseous end products. High-pressure liquid chromatographic analyses of CuO oxidation products of sound and degraded pine lignin indicated that no substantial chemical modifications of the remaining lignin polymer, such as demethoxylation and dearomatization, occurred during biodegradation. The polysaccharide components of pine lignocellulose and purified cellulose were relatively rapidly mineralized to methane and carbon dioxide; 31 to 37% of the pine polysaccharides and 56 to 63% of the purified cellulose were recovered as labeled gaseous end products. An additional 10 to 20% of the polysaccharide substrates was recovered as dissolved degradation products. Overall, these results indicate that elevated temperatures can greatly enhance rates of anaerobic degradation of lignin and lignified substrates to methane and low-molecular-weight aromatic compounds.     

Contribution to lignocellulose degradation and DOC formation from a salt marsh macrophyte by the ascomycete Phaeosphaeria spartinicola

Abstract Specifically radiolabeled [¹⁴C-lignin] lignocellulose and uniformly [U-¹⁴C] lignocellulose from the salt marsh grass Spartina alterniflora were incubated with the ascomycete Phaesphaeria s spartinicola . This fungus is the predominant one found on decaying standing dead S. alterniflora leaves in the salt marsh ecosystem. After 45 days of incubation at 20°C, 3.3% of the lignin moiety was mineralized to ¹⁴CO₂ and 2.7% solubilized to DO¹⁴C. Mineralization of the polysaccharides was seven times faster than that of the lignin. About 22% of the radioactivity was evolved as ¹⁴CO₂ but merely 4% was solubilized to DO¹⁴C within the incubation time. Experiments monitoring the ergosterol content of the mycelium incubated with S. alterniflora plant material were done to elucidate the carbon conversion efficiency of the fungus as well as the influence of the cinnamyl phenols p -coumaric and ferulic acid on lignocellulose degration. After 21 days of incubation, P. spartinicola showed a growth yield of 0.45 and 0.38 with and without the additional cinnamyl phenols, respectively. Grown on unextracted S. alterniflora the fungus caused a loss of organic plant material of about 50% with a corresponding growth yield of 0.38 during the incubation period. Investigation of cupric oxide oxidation products of sound and degraded lignocellulose revealed a preferential utilization of the syrinyl and cinnamly phenols compared with vanilly phenols.     

Degradation of Softwood, Hardwood, and Grass Lignocelluloses by Two Streptomyces Strains

Two Streptomyces strains, S. viridosporus T7A and S. setonii 75Vi2, were grown on softwood, hardwood, and grass lignocelluloses, and lignocellulose decomposition was followed by monitoring substrate weight loss, lignin loss, and carbohydrate loss over time. Results showed that both Streptomyces strains substantially degraded both the lignin and the carbohydrate components of each lignocellulose; however, these actinomycetes were more efficient decomposers of grass lignocelluloses than of hardwood or softwood lignocelluloses. In particular, these Streptomyces strains were more efficient decomposers of grass lignins than of hardwood or softwood lignins.     

Degradation of extractive-free lignocelluloses by Coriolus versicolor and Poria placenta

Summary The wood-decay fungi Coriolus versicolor, a white-rot fungus, and Poria placenta, a brown-rot fungus, were grown on an extractive-free lignocellulose prepared from quackgrass (Agropyron repens). Their abilities to decompose this lignocellulose were compared to their abilities to decompose softwood (Picea pungens) and hardwood (Acer rubrum) lignocelluloses. The two fungi were grown on malt-extract dampened lignocelluloses at 28°C for up to 12 weeks. Replicate cultures were periodically harvested and lignocellulose decomposition was followed by monitoring substrate weight loss, lignin loss, and carbohydrate loss. Coriolus versicolor decomposed the lignin and carbohydrate components of the grass lignocellulose as efficiently as the softwood and hardwood lignocelluloses. Poria placenta, however, was not an efficient degrader of either lignin or carbohydrate in the grass lignocellulose. Poria placenta readily decomposed carbohydrate components of the softwood lignocellulose but not the hardwood lignocellulose.Paper number 81520 of the Idaho Agricultural Experiment Station     

Lignocellulose Mineralization by Arctic Lake Sediments in Response to Nutrient Manipulation

Mineralization of specifically labeled ¹⁴C-cellulose- and ¹⁴C-lignin-labeled lignocelluloses by Toolik Lake, Alaska, sediments was examined in response to manipulation of various environmental factors. Mineralization was measured by quantifying the amount of labeled CO₂ released from the specifically labeled substrates. Nitrogen (NH₄NO₃) and, to a greater degree, phosphorus (PO₄⁻³) additions enhanced the mineralization of white pine (Pinus strobus) cellulose during the summer of 1978. Nitrogen and phosphorus together had no cumulative effect. During the summer of 1979, nitrogen or phosphorus alone had only a slight stimulatory effect on the mineralization of a sedge (Carex aquatilis) cellulose; however, together, they had a dramatic effect. This variable response of mineralization to nutrient addition between 1978 and 1979 was probably attributable to year-to-year variation in nutrient availability within the lake. Cellobiose addition and oxygen depletion inhibited the amount of pine cellulose mineralized. Whereas addition of nitrogen to oxygen-depleted treatments had limited effect, addition of phosphorus resulted in mineralizations equal to or greater than that of the controls. Nitrogen had no effect on mineralization of pine or Carex lignins. Phosphorus, however, inhibited mineralization of both lignins. With Carex lignin, the phosphorus inhibition occurred at a concentration as low as 0.1 μM. The antagonistic role of phosphorus in cellulose and lignin mineralizations may be of significance in understanding the increased proportion of lignin relative to cellulose in decomposing litter.     

Fungal decomposition of Abies needle and Betula leaf litter

The effect of litter type and incubation temperature on the ability of fungi to decompose leaf litter of subalpine trees was examined by a pure-culture test. Mass loss of Abies needle and Betula leaf litter and utilization patterns of lignin and carbohydrates were investigated under two temperature conditions (20 C and 10 C) and compared for 29 species in basidiomycetes, ascomycetes and zygomycetes. The decomposing ability was generally higher in basidiomycetes than in ascomycetes and zygomycetes. Mass loss (% original mass) of litter was higher in Betula than in Abies and higher at 20 C than at 10 C. The 29 fungi were divided into lignocellulose decomposers, cellulose decomposers and sugar fungi based on their substrate utilization in Abies and Betula litter. Mass loss of lignin and carbohydrates by lignocellulose and cellulose decomposers was higher in Betula than in Abies. Mass loss of carbohydrates was higher at 20 C than at 10 C, but the temperature did not influence mass loss of lignin, indicating lignin decomposition by fungi was less sensitive to temperature than carbohydrate decomposition. Lignin/carbohydrate loss ratio (L/C) of Collybia spp. that caused selective delignification was lower at 20 C than at 10 C. These results indicate that the decomposability of litter, lignin and carbohydrate was different between Abies and Betula and that temperature affected not only the rate at which fungi decompose litter but also the ability of fungi to use lignin and carbohydrates.     

Lignin Degradation by Streptomyces viridosporus: Isolation and Characterization of a New Polymeric Lignin Degradation Intermediate

A new, quantitatively significant intermediate formed during lignin degradation by Streptomyces viridosporus T7A was isolated and characterized. In Streptomyces-inoculated cultures, the intermediate, an acid-precipitable, polyphenolic, polymeric lignin (APPL), accumulated in the growth medium. The APPL was a water-soluble polymer probably consisting of a heterogeneous mixture of molecular weight components of ≥20,000. APPLs were precipitable from culture filtrates after they had been acidified to pH <3 to 5. Noninoculated controls yielded little APPL, but supernatant solutions from inoculated cultures produced quantities of APPL that correlated with the biodegradability of the lignocellulose type. Maximal recovery of APPL was obtained from corn lignocellulose, reaching 30% of the initial lignin present in the substrate. APPLs contained small amounts of carbohydrate, organic nitrogen, and inorganic materials. The lignin origin of APPLs was confirmed by chemical analyses, which included acidolysis, permanganate oxidation, elemental analyses, functional group analyses, nuclear magnetic resonance spectroscopy, and ¹⁴C isotopic techniques. Analyses of APPLs from corn lignocelluloses showed that S. viridosporus-degraded APPLs were lignin derived but significantly different in structure from APPLs derived from uninoculated controls or from a standard corn milled-wood lignin. Degraded APPLs were enriched in phenolic hydroxyl groups and, to a small extent, in carboxyl groups. Degradative changes appeared to be largely oxidative and were thought to involve substantial cleavage of p-hydroxy ether linkages and methoxyl groups in the lignin.     

Effects of Selected Assay Parameters on Measurement of Lignocellulose Mineralization with a Radiolabeled Substrate

Varying the amount of labeled substrate or the amount of leaf material resulted in significant nonlinear changes in lignocellulose mineralization, as measured with natural [¹⁴C]lignin-labeled lignocellulose. The use of periodic rather than continuous aeration was found not to have significant effects on measured mineralization.     

Microbial Decomposition of Wood in Streams: Distribution of Microflora and Factors Affecting [14C]Lignocellulose Mineralization

The distribution and lignocellulolytic activity of the microbial community was determined on a large log of Douglas fir (Pseudotsuga menziesii) in a Pacific Northwest stream. Scanning electron microscopy, plate counts, and degradation of [¹⁴C]lignocelluloses prepared from Douglas fir and incubated with samples of wood taken from the surface and within the log revealed that most of the microbial colonization and lignocellulose-degrading activity occurred on the surface. Labeled lignocellulose and surface wood samples were incubated in vitro with nutrient supplements to determine potential limiting factors of [¹⁴C]lignocellulose degradation. Incubations carried out in a nitrogenless mineral salts and trace elements solution were no more favorable to degradation than those carried out in distilled water alone. Incubations supplemented with either (NH₄)₂SO₄ or organic nitrogen sources showed large increases in the rates of mineralization over incubations with mineral salts and trace elements alone, with the greatest effect being observed from an addition of (NH₄)₂SO₄. Subsequent incubations with (NH₄)₂SO₄, KNO₃, and NH₄NO₃ revealed that KNO₃ was the most favorable for lignin degradation, whereas all three supplements were equally favorable for cellulose degradation. Supplementation with glucose repressed both lignin and cellulose mineralization. The results reported in this study indicate that nitrogen limitation of wood decomposition may exist in streams of the Pacific Northwest. The radiotracer technique was shown to be a sensitive and useful tool for assessing relative patterns of lignocellulose decay and microbial activity in wood, along with the importance of thoroughly characterizing the experimental system before its general acceptance.