Patterns of amino acid utilization by aquatic hyphomycetes

Summary The utilization of amino acids in leaf protein and leaf leachate by aquatic hyphomycetes was studied during decomposition in a combined field and laboratory experiment. Leaves were sampled from a stream which exhibited a seasonal variation in free amino acid concentration in surface water, reaching peaks in autumn and winter. In the leaf drift environment the concentration of amino acids was approximately two orders of magnitude higher than in surface water. Protein amino acid content, which was higher in alder leaves than in beech leaves, decreased exponentially and faster in alder leaves, so that protein amino acid content was similar in the two leaf types after 9–10 weeks decomposition. From 55% to 75% of leaf amino acids were used instantaneously by attached fungi, which grew well, especially on alder leaves, regardless of the presence of a grazing amphipod. If nitrogen was a limiting nutrient source for fungi, it appeared to be more advantageous to colonize alder leaves. Four times more fungal species were found on alder leaves than on beech leaves. The changes in concentration of amino acids in leaves and water was described by a set of differential equations. Rate constants for the transfer of amino acids from leaves and water were estimated from experimental data and the preference in fungi for protein-bound and free amino acids evaluated.The amounts of free amino acids in water absorbed by fungi varied between leaf types and leaves at different stages of decay. Experimental data showed a switching behaviour in fungal absorption of dissolved amino acids so that absorption became superproportional at a certain proportion of free amino acids available in the water.     

Autumnal changes in total nitrogen, salt-extractable proteins and amino acids in leaves and adjacent bark of black alder, eastern cottonwood and white basswood

Autumnal changes in total nitrogen, salt-extractable protein and amino acid concentrations in leaves and adjacent bark of black alder [ Ainus glutinosa (L.) Gaertn.], eastern cottonwood ( Populus deltoides Bartr. ex Marsh.) and white basswood ( Tilia heterophylla Vent.) were determined for trees growing on minespoils and a prairiederived loamy soil in central Illinois. The composition of free amino acids in foliage was also determined at peak concentration for each tree species during late senescence. Total nitrogen concentration in the leaves decreased slowly throughout most of the fall for all species. In the final stages of senescence, total leaf nitrogen concentrations were about halved in eastern cottonwood and white basswood but continued to decrease slowly in black alder. The concentration of salt-extractable proteins in leaves of all species peaked early in the fall and then declined prior to leaf abscission. This decline coincided with an increase in the concentration of free amino acids in the leaves. The increase stabilized in both eastern poplar and white basswood but continued in black alder. Glutamine in black alder and eastern cottonwood, and asparagine in white basswood were the most abundant free amino acids at the time of peak concentration of total free amino acids in senescent leaves. Bark of trees of all species had higher nitrogen concentrations and higher proportions of salt-extractable proteins to estimated total proteins after leaf senescence than during the preceding summer. Results indicate that autumnal fluxes in leaf and bark nitrogen fractions of alder can differ substantially from fluxes in other broadleaved winter-deciduous trees in a way which suggests that alder does not effectively conserve leaf nitrogen through retranslocation to bark tissue.     

Impacts of invasive riparian knotweed on litter decomposition,aquatic fungi,and macroinvertebrates

Bohemian knotweed (Polygonum × bohemicum), the hybrid between Japanese and giant knotweed, is the most common invasive knotweed species in western North America and the most difficult to control. Invasive knotweed congeners spread aggressively along streams and establish dense monotypic stands, reducing riparian plant species diversity. Allochthonous organic matter inputs from riparian plants are an important source of energy and nutrients for organisms in small streams. However, little information exists concerning the influence of knotweed on stream processes. This study examines the quality of Bohemian knotweed leaves compared to native red alder and black cottonwood leaves, along with leaf-associated fungal biomass accumulation, macroinvertebrate communities, and decay rates from three forested streams in western Washington State. Senesced knotweed leaves were lower in nitrogen and phosphorus, and higher in cellulose, fiber, and lignin content than alder leaves, but were more similar to cottonwood leaves. Fungal biomass differed among species and changed over time. Macroinvertebrate shredders collected from leaf packs after 31 days were proportionately more abundant on alder leaves than knotweed and cottonwood. Decay rates were not significantly different among leaf species, but during the first 31 days alder broke down faster than knotweed. After 56 days, all of the leaf packs were mostly decomposed. Overall, these findings do not show major discrepancies between leaf species except those related to initial litter structural and chemical quality. However, changes in the timing and quantity of litter inputs are also important factors to be considered in understanding the impact of invasive knotweed on stream ecosystem processes.     

Fungi associated with the aerial parts of Malaysian mangrove plants

The leaf surface fungi associated with nine species of mangrove plants includingAvicennia alba, A. officinalis, Bruguiera parviflora, Ceriops tagal, Rhizophora apiculata, R. mucronata, Sesuvium portulacastrum, Sonneratia alba, andXylocarpus mollucensis were studied using direct observation techniques and leaf washings. Over 40 fungal taxa were isolated from the leaf washings. Of these, species ofAspergillus, Choanephora, Cladosporium, Curvularia, Fusarium, Nigrospora, Penicillium, Pestalotiopsis, Trichoderma, andZygosporium were frequently encountered in the washings of all nine mangrove plants. Fewer species of fungi includingCladosporium oxysporum, Corynespora cassiicola, Fusarium, Penicillium, Pestalotiopsis, andZygosporium were capable of growth on the washed leaves. The major phylloplane fungus on plants with higher leaf tannin content (e.g.B. parviflora, C. tagal, Rhizophora spp., andX. mollucensis) wasPestalotiopsis. Leaves with relatively lower amounts of tannin supported the proliferation ofFusarium as the major fungus. Fungi were present on the plumule and cotyledonous sheath even before the leaves opened.Pestalotiopsis persisted throughout the development and growth of the leaves. Many of the fungi encountered on senescent leaves have been reported in earlier studies to be the primary colonisers of submerged decaying leaves.     

Comparison of Degradative Ability, Enzymatic Activity, and Palatability of Aquatic Hyphomycetes Grown on Leaf Litter

Stream fungi have the capacity to degrade leaf litter and, through their activities, to transform it into a more palatable food source for invertebrate detritivores. The objectives of the present study were to characterize various aspects of fungal modification of the leaf substrate and to examine the effects these changes have on leaf palatability to detritivores. Fungal species were grown on aspen leaves for two incubation times. Leaves were analyzed to determine the weight loss, the degree of softening of the leaf matrix, and the concentrations of ATP and nitrogen associated with leaves. The activities of a protease and 10 polysaccharide-degrading enzymes produced by each fungus were also determined. Most fungi caused similar changes in physicochemical characteristics of the leaves. All fungi exhibited the capability to depolymerize pectin, xylan, and cellulose. Differences among fungi were found in their capabilities to produce protease and certain glycosidases. Leaf palatability was assessed by offering leaves of all treatments to larvae of two caddisfly shredders (Trichoptera). Feeding preferences exhibited by the shredders were similar and indicated that they perceived distinct differences among fungi. Two fungal species were highly consumed, some moderately and others only slightly. No relationships were found between any of the fungal characteristics measured and detritivore feeding preferences. Apparently, interspecific differences among fungi other than parameters associated with biomass or degradation of structural polysaccharides influence fungal palatability to caddisfly detritivores.     

Riparian plant species loss alters trophic dynamics in detritus-based stream ecosystems

Riparian vegetation is closely connected to stream food webs through input of leaf detritus as a primary energy supply, and therefore, any alteration of plant diversity may influence aquatic ecosystem functioning. We measured leaf litter breakdown rate and associated biological parameters in mesh bags in eight headwater streams bordered either with mixed deciduous forest or with beech forest. The variety of leaf litter types in mixed forest results in higher food quality for large-particle invertebrate detritivores (‘shredders’) than in beech forest, which is dominated by a single leaf species of low quality. Breakdown rate of low quality (oak) leaf litter in coarse mesh bags was lower in beech forest streams than in mixed forest streams, a consequence of lower shredder biomass. In contrast, high quality (alder) leaf litter broke down at similar rates in both stream categories as a result of similar shredder biomass in coarse mesh bags. Microbial breakdown rate of oak and alder leaves, determined in fine mesh bags, did not differ between the stream categories. We found however aquatic hyphomycete species richness on leaf litter to positively co-vary with riparian plant species richness. Fungal species richness may enhance leaf litter breakdown rate through positive effects on resource quality for shredders. A feeding experiment established a positive relationship between fungal species richness per se and leaf litter consumption rate by an amphipod shredder (Gammarus fossarum). Our results show therefore that plant species richness may indirectly govern ecosystem functioning through complex trophic interactions. Integrating microbial diversity and trophic dynamics would considerably improve the prediction of the consequences of species loss.     

Fungal community composition shifts along a leaf degradation gradient in a European beech forest

Aims

The fungal communities in living and decomposed leaves of European Beech (Fagus sylvatica) were compared to identify the phyllosphere fungi involved in litter decomposition at a site in Bavaria, Germany.

Methods

New primers were designed to cover a broad range of fungal ribosomal DNA sequence diversity. Following ‘environmental PCR’, clone libraries from each of five samples of living leaves (surface-sterilized and untreated), freshly fallen, initially and highly decomposed leaves, were screened using RFLP fingerprinting.

Results

Statistical analysis (ANOSIM) revealed that the fungal communities colonizing living (a) and initially decomposed leaves (c) significantly differed between each other and from freshly fallen (b) and highly decomposed leaves (d). Fungal assemblages of a and d were statistically indistinguishable from each other and from the endophyllous fungal community in living leaves.

Conclusions

The results showed that endophyllous fungi play a role throughout the whole decomposition process of beech leaf litter. Therefore, clarification of the life cycle of certain endophytic and/or soil fungi may only be achieved by considering both phyllosphere and soil habitats.     

Population genetics of the aquatic fungus Tetracladium marchalianum over space and time

Aquatic hyphomycete fungi are fundamental mediators of energy flow and nutrient spiraling in rivers. These microscopic fungi are primarily dispersed in river currents, undergo substantial annual fluctuations in abundance, and reproduce either predominantly or exclusively asexually. These aspects of aquatic hyphomycete biology are expected to influence levels and distributions of genetic diversity over both spatial and temporal scales. In this study, we investigated the spatiotemporal distribution of genotypic diversity in the representative aquatic hyphomycete Tetracladium marchalianum. We sampled populations of this fungus from seven sites, three sites each in two rivers in Illinois, USA, and one site in a Wisconsin river, USA, and repeatedly sampled one population over two years to track population genetic parameters through two seasonal cycles. The resulting fungal isolates (N = 391) were genotyped at eight polymorphic microsatellite loci. In spite of seasonal reductions in the abundance of this species, genotypic diversity was consistently very high and allele frequencies remarkably stable over time. Likewise, genotypic diversity was very high at all sites. Genetic differentiation was only observed between the most distant rivers (∼450 km). Clear evidence that T. marchalianum reproduces sexually in nature was not observed. Additionally, we used phylogenetic analysis of partial β-tubulin gene sequences to confirm that the fungal isolates studied here represent a single species. These results suggest that populations of T. marchalianum may be very large and highly connected at local scales. We speculate that large population sizes and colonization of alternate substrates in both terrestrial and aquatic environments may effectively buffer the aquatic populations from in-stream population fluctuations and facilitate stability in allele frequencies over time. These data also suggest that overland dispersal is more important for structuring populations of T. marchalianum over geographic scales than expected.     

Effect of Oxygen Concentration on C-Photoassimilate Transport from Leaves of Salvia splendens L

Partitioning and transport of recently fixed photosynthate was examined following ¹⁴CO₂ pulse-labeling of intact, attached leaves of Salvia splendens L. maintained in an atmosphere of 300 microliters per liter CO₂ and 20, 210, or 500 milliliters per liter O₂. Under conditions of increasing O₂ (210, 500 milliliters per liter), a smaller percentage of the recently fixed ¹⁴C in the leaf was allocated to starch, whereas a greater percentage of the fixed ¹⁴C appeared in amino acids, particularly serine. The increase in ¹⁴C in amino acids was reflected in material exported from source leaves. A higher percentage of ¹⁴C in serine, glycine, and glutamate was recovered in petiole extracts when source leaves were maintained under elevated O₂ levels. Although pool sizes of these amino acids were increased in both the leaves and petioles with increasing photorespiratory activity, no significant changes in either ¹⁴C distribution or concentration of transport sugars (i.e. stachyose, sucrose, verbascose) were observed. The data indicate that, in addition to being recycled intracellularly into Calvin cycle intermediates, amino acids produced during photorespiration may also serve as transport metabolites, allowing the mobilization of both carbon and nitrogen from the leaf under conditions of limited photosynthesis.     

Species diversity of freshwater hyphomycetes in some streams of Pakistan. III. Autumnal colonization of freshwater hyphomycetes on bait leaves

Significantly more species of freshwater hyphomycetes colonized bait leaves of alder (16 species) and willow (14 species) during early submersion period (1 wk) than those of oak (8 species). Production of conidia was also higher on alder and willow leaves than on oak leaves. Higher amounts of dry mass were lost from the alder (28.6%) and willow leaves (29.4%) than from the oak leaves (18.7%). Rapid loss of dry mass was accompanied by rapid development of a fungal community. Fungal species took a longer time (6 wk) to reach their peak of occurrence on oak than on willow and alder leaves (3–4 wk).Flagellospora curvula dominated the assemblages of freshwater hyphomycetes on alder and willow leaves, andLunulospora curvula dominated the assemblage on oak leaves. The freshwater hyphomycete community showed higher values of species diversity (H value) on oak leaves than on alder and willow leaves.     

The breakdown and decomposition of sweet chestnut (Castanea sativa mill.) and beech (Fagus sylvatica L.) leaf litter in two deciduous woodland soils

Summary Changes in the carbon, hydrogen, nitrogen and polyphenol content of chestnut and beech leaves were measured during the first year after fall.Chestnut leaves had an initial carbon, hydrogen and nitrogen content (by weight) of 48.71%, 5.56% and 0.77% respectively; beech had a similar carbon and hydrogen content (47.77% and 5.36%) but less nitrogen (0.56%).Both leaf litter types showed percentage increases in nitrogen content during the study period but only the beech showed absolute increases in nitrogen content of up to 66.7% of the initial weight present in the leaves. The percentage increases in the nitrogen content of chestnut litter were largely attributable to more rapid losses of non-nitrogenous leaf constituents while the weight of nitrogen present in the leaves remained relatively constant.The percentage carbon and hydrogen contents of the chestnut and beech litter showed changes of less than 1% throughout the year, indicating that carbohydrate losses were directly proportional to weight losses.Soil animals fed on chestnut leaves to a far greater extent than beech leaves; this difference did not appear to be directly attributable to differences between the nitrogen contents or C/N ratios of the two leaf litter species. The gross polyphenol contents of chestnut and beech litter showed an intraspecific inverse correlation with the feeding activities of soil animals on the leaves, but did not account for interspecific differences in leaf palatability since chestnut leaves were eaten when they contained higher polyphenol concentrations than beech leaves. However, there was an interspecific negative correlation between palatability and the presence of protocatechuic and gallic acids.     

Effects of Leaf Litter Species on Macroinvertebrate Colonization during Decomposition in a Portuguese Stream

This study evaluated if there are differences in leaf breakdown and invertebrate colonization among tree species differing in quality (toughness), and which factors could influence these differences. Common alder leaves decomposed significantly faster then either sweet chestnut or Spanish oak (k values of –0.0332, –0.0108, and –0.0112, respectively) during the first 2 months. Shredder abundance was highest when leaf mass remaining was 50%, and the samples clustered in mixed groups of sampling dates and leaf species, suggesting that stage of decomposition was an important factor influencing shredder colonization. During the first two months of decomposition, the physicochemical characteristics of leaf litter and the interaction between leaf toughness and the occurrence of frequent spates seemed to be the main factors affecting leaf breakdown rates in the stream. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Effects of Riparian Plant Diversity Loss on Aquatic Microbial Decomposers Become More Pronounced with Increasing Time

We examined the potential long-term impacts of riparian plant diversity loss on diversity and activity of aquatic microbial decomposers. Microbial assemblages were obtained in a mixed-forest stream by immersion of mesh bags containing three leaf species (alder, oak and eucalyptus), commonly found in riparian corridors of Iberian streams. Simulation of species loss was done in microcosms by including a set of all leaf species, retrieved from the stream, and non-colonized leaves of three, two or one leaf species. Leaves were renewed every month throughout six months, and microbial inoculum was ensured by a set of colonized leaves from the previous month. Microbial diversity, leaf mass loss and fungal biomass were assessed at the second and sixth months after plant species loss. Molecular diversity of fungi and bacteria, as the total number of operational taxonomic units per leaf diversity treatment, decreased with leaf diversity loss. Fungal biomass tended to decrease linearly with leaf species loss on oak and eucalyptus, suggesting more pronounced effects of leaf diversity on lower quality leaves. Decomposition of alder and eucalyptus leaves was affected by leaf species identity, mainly after longer times following diversity loss. Leaf decomposition of alder decreased when mixed with eucalyptus, while decomposition of eucalyptus decreased in mixtures with oak. Results suggest that the effects of leaf diversity on microbial decomposers depended on leaf species number and also on which species were lost from the system, especially after longer times. This may have implications for the management of riparian forests to maintain stream ecosystem functioning.     

Colonization of Japanese beech leaves by phyllosphere fungi

To examine the colonization of living leaves from buds and twigs by phyllosphere fungi of Japanese beech, the mycobiota were investigated on buds and twigs and on leaves covered with well-ventilated bags before budbreak. Ten phyllosphere fungi were isolated from rolled-up leaves within buds, bud scales, and twigs. However, frequencies of phyllosphere fungi on expanded leaves were reduced markedly when the buds were covered with well-ventilated bags before budbreak compared to the leaves that were not covered. This observation suggests that invasion of the fungi to the leaves from buds and twigs may be possible but is not the main route. Horizontal transmission may be common in endophytes and epiphytes of beech leaves. Phyllosphere mycobiota were then compared between sun and shade leaves. Of 13 species recorded as phyllosphere fungi, the frequencies of 2 species were lower and those of 3 species were higher in sun leaves than in shade leaves. Frequencies of the other 8 phyllosphere species were not different between sun and shade leaves. This result indicates that the colonization of leaves by some phyllosphere fungi was affected by the microenvironmental conditions on leaf surfaces.     

Fungi associated with the aerial parts of Malaysian mangrove plants

The leaf surface fungi associated with nine species of mangrove plants includingAvicennia alba, A. officinalis, Bruguiera parviflora, Ceriops tagal, Rhizophora apiculata, R. mucronata, Sesuvium portulacastrum, Sonneratia alba, andXylocarpus mollucensis were studied using direct observation techniques and leaf washings. Over 40 fungal taxa were isolated from the leaf washings. Of these, species ofAspergillus, Choanephora, Cladosporium, Curvularia, Fusarium, Nigrospora, Penicillium, Pestalotiopsis, Trichoderma, andZygosporium were frequently encountered in the washings of all nine mangrove plants. Fewer species of fungi includingCladosporium oxysporum, Corynespora cassiicola, Fusarium, Penicillium, Pestalotiopsis, andZygosporium were capable of growth on the washed leaves. The major phylloplane fungus on plants with higher leaf tannin content (e.g.B. parviflora, C. tagal, Rhizophora spp., andX. mollucensis) wasPestalotiopsis. Leaves with relatively lower amounts of tannin supported the proliferation ofFusarium as the major fungus. Fungi were present on the plumule and cotyledonous sheath even before the leaves opened.Pestalotiopsis persisted throughout the development and growth of the leaves. Many of the fungi encountered on senescent leaves have been reported in earlier studies to be the primary colonisers of submerged decaying leaves.     

Non-protein amino acids of Acacia species and their effect on the feeding of the acridids Anacridium melanorhodon and Locusta migratoria

The leaves of Acacia species have been found to contain homoarginine, pipecolic acid and 4-hydroxy-pipecolic acid. The nymphs of the tree locust Anacridium melanorhodon, which feed on the leaves of Acacia species, were not inhibited from feeding on palatable media containing concentrations of these amino acids equivalent to, or greater than, those found in the leaves. The graminivorous Locusta migratoria was more sensitive to these compounds, inhibitory effects being observed at concentrations comparable to those found in the leaves. The inhibitory effects of mixtures of homoarginine and pipecolic acid were additive in A. melanorhodon but not in L. migratoria. Three of the non-protein amino acids found in the seeds of Acacia species, 2,3-diaminopropionic acid, 2-amino-3-acetylaminopropionic acid and 2-amino-3-oxalylaminopropionic acid, were more effective inhibitors of feeding in Anacridium than were the leaf amino acids.     

Interactions between fungi,bacteria and beech leaves in a stream microcosm

Summary Immigration and colonization of isolates of naturally occurring stream bacteria and hyphomycetes on beech leaves were studied in a laboratory stream microcosm. Fungal spores were more successful immigrants, especially on new leaves, than bacteria, which were more repelled than attracted by the substrate. Fewer bacteria immigrated to older leaves than to new, and bacteria multiplied faster in water than on leaves. Fungi and bacteria showed synergistic relationships so that each group grew significantly faster in presence of the other group. If one considers, differences in immigration, colonization and synergism patterns, fungal mycelia doubled about 10 times faster than bacterial cells which might explain the dominance of fungi usually found on leaves in early decay. The individual fungal species could be assigned to one of three colonization groups; one of fugitive species, preceding a second group of species that grew from rarity to dominance, and a third group of very slow colonizers. The leachate was fractionated in different molecular size classes by gel chromatography, and the fraction around 2500D in the new leaf leachate was associated with a high concentration of polyphenols. High-pressure liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) revealed the presence of 16 phenolic acids in the new leaf leachate at concentrations ranging from < 1 to 640 g l^–1. All fungi except the species with the slowest growth rate grew faster on leachate with the fraction around 2500D removed, and the density of bacteria was significantly reduced when pure stream water was supplemented with compounds from the same fraction.     

Do aphid galls provide good nutrients for the aphids?: Comparisons of amino acid concentrations in galls among Tetraneura species (Aphididae: Eriosomatinae)

Some aphid species induce leaf galls, in which the fundatrix parthenogenetically produces many nymphs. In order to ensure high performance, galls have to provide the aphids with sufficient nutrients, in particular, amino acids as a nitrogen source. We tested this hypothesis using six Tetraneura aphid species that induce closed galls. We extracted free amino acids from the whole gall tissues of unit weight and quantified the concentration of each amino acid. There were large differences in the total amino acid concentrations among galls of the Tetraneura species. Tetraneura species in which higher concentrations of total amino acids were found in the gall tended to produce larger numbers of offspring. Of the amino acids found, asparagine was predominant in the gall. The asparagine concentration in T. yezoensis galls was several hundred times as high as in control leaves. We discussed why such a high level of asparagine accumulates in aphid galls.     

Essential oils of four Myrtaceae species from the Brazilian southeast

The essential oils of the leaves of Eugenia acutata, Eugenia candolleana, Eugenia copacabanensis and Myrcia splendens (Myrtaceae) from Brazil’s southeastern Atlantic Forest were obtained by hydrodistillation and analyzed by GC–MS. Oxygenated sesquiterpenes were predominant in E. copacabanensis (54.3%) and E. candolleana (50.9%) whilst hydrocarbon species predominated in E. acutata (83.4%) and M. splendens (94.5%). trans-Caryophyllene was the most abundant component in E. acutata. Isomers of guaiol and cadinol alcohols, followed by δ-elemene and viridiflorene, were the major components of the essential oil of the leaves of E. candolleana. Hydrocarbons and alcohols of the cadinane-type predominated in E. copacabanensis the most abundant being epi-cubenol (14%). M. splendens had 80% α-bisabolene in the leaf oil along with <5% β-farnesene. Additionally, E. copacabanensis exhibited 13.7% monoterpenes. Whereas the bisabolene-rich M. splendens oil is highly similar to that of other Myrcia species reported elsewhere, the Eugenia species oils corroborated the complex array and differing abundances of terpene classes within this genus. This study generated data which may provide further comprehension of the phylogenetic relationships between Myrtaceae genera and species.