Colonization and succession of fungi during decomposition of Swida controversa leaf litter

Decomposition processes of Swida controversa leaves were investigated in initially sterilized (fungi-excluded) and nonsterilized freshly fallen leaves to examine the relationship between chemical changes and fungal succession during decomposition and the effect of exclusion of previously established phyllosphere fungi from leaves on subsequent decomposition and fungal succession. Fifteen species were isolated frequently from decomposing leaves with surface-disinfection and washing methods. These fungi were divided into early and late colonizers according to their occurrence during decomposition. The 1.5 y decomposition process was divided into three stages characterized by different dominant organic chemical constituents. A clear relationship was demonstrated between chemical changes and fungal succession. Total hyphal length and frequencies of some early colonizers were reduced in initially sterilized leaves at 3 wk, but this had no significant effect on loss of litter mass or chemical changes during the first 3 wk or on the subsequent decomposition and fungal succession.     

Determining diversity of freshwater fungi on decaying leaves: comparison of traditional and molecular approaches

Traditional microscope-based estimates of species richness of aquatic hyphomycetes depend upon the ability of the species in the community to sporulate. Molecular techniques which detect DNA from all stages of the life cycle could potentially circumvent the problems associated with traditional methods. Leaf disks from red maple, alder, linden, beech, and oak as well as birch wood sticks were submerged in a stream in southeastern Canada for 7, 14, and 28 days. Fungal biomass, estimated by the amount of ergosterol present, increased with time on all substrates. Alder, linden, and maple leaves were colonized earlier and accumulated the highest fungal biomass. Counts and identifications of released conidia suggested that fungal species richness increased, while community evenness decreased, with time (up to 11 species on day 28). Conidia of Articulospora tetracladia dominated. Modifications of two molecular methods-denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) analysis-suggested that both species richness and community evenness decreased with time. The dominant ribotype matched that of A. tetracladia. Species richness estimates based on DGGE were consistently higher than those based on T-RFLP analysis and exceeded those based on spore identification on days 7 and 14. Since traditional and molecular techniques assess different aspects of the fungal organism, both are essential for a balanced view of fungal succession on leaves decaying in streams.     

Determining Diversity of Freshwater Fungi on Decaying Leaves: Comparison of Traditional and Molecular Approaches

Traditional microscope-based estimates of species richness of aquatic hyphomycetes depend upon the ability of the species in the community to sporulate. Molecular techniques which detect DNA from all stages of the life cycle could potentially circumvent the problems associated with traditional methods. Leaf disks from red maple, alder, linden, beech, and oak as well as birch wood sticks were submerged in a stream in southeastern Canada for 7, 14, and 28 days. Fungal biomass, estimated by the amount of ergosterol present, increased with time on all substrates. Alder, linden, and maple leaves were colonized earlier and accumulated the highest fungal biomass. Counts and identifications of released conidia suggested that fungal species richness increased, while community evenness decreased, with time (up to 11 species on day 28). Conidia of Articulospora tetracladia dominated. Modifications of two molecular methods—denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) analysis—suggested that both species richness and community evenness decreased with time. The dominant ribotype matched that of A. tetracladia. Species richness estimates based on DGGE were consistently higher than those based on T-RFLP analysis and exceeded those based on spore identification on days 7 and 14. Since traditional and molecular techniques assess different aspects of the fungal organism, both are essential for a balanced view of fungal succession on leaves decaying in streams.     

Fungal successions on pine needles fallen at different seasons: The succession of interior colonizers

Experimental studies were carried out to investigate seasonal effects on the fungal succession in the interior or decaying pine needles. At different seasons, the needles fallen for a short period were collected and marked, then placed on the surface of the O horizon in a pine forest. The needles were removed at intervals and their interior fungal communities were examined by using a surface sterilization technique. The successions of interior colonizers observed on the fallen needles at four different times are roughly divided into three groups based on the composition of species colonizing from litter. Seasonal shifts in the species combination were discussed with climatic and biotic factors. As a result, temperature at the surface of litter appeared to be a cardinal factor contributing to these seasonal changes in the succession of interior colonizers. Contributions to Sugadaira Montane Research Center, No. 164.     

Dematiaceous hyphomycetes inhabiting decaying blackish needles ofAbies firma and their distribution in the Kanto district, Japan

Dematiaceous mitosporic fungi darkening decaying fir needles on the ground were studied. Fungal communities on decaying, blackish fir needles were investigated in nine sites of the Kanto district, Japan, using a washing method. A total 108 taxa was recorded from 540 sampled needles. Among abundantly occurring dematiaceous fungi,Anungitea continua, A. uniseptata andEndophragmiella uniseptata were recognized as the major colonizers, forming a hyphal network on the surface of decaying fallen needles and darkening them. The effects of climate on the distributions of seven dematiaceous fungi included the major colonizers were analyzed. The abundance (proportion of needles colonized by a fungal species) ofChaetopsina fulva showed a significantly positive correlation with annual mean air temperature at each sampling site. No other significant correlations between the selected climatic factors and the distributions of dematiaceous fungi were recognized. Contribution No. 176 from Sugadaira Montane Research Center, University of Tsukuba.     

Fungal successions on pine needles fallen at different seasons: The succession of surface colonizers

Field experiments were carried out to investigate influences of seasonal change on the fungal succession occurring on the surface of decaying pine needles at a moder site in Japan. At different seasons, the needles fallen for a short period were collected and marked, then placed on the surface of the O horizon. The needles were removed at intervals and their fungal communities were examined by using a washing technique. Unlike the successions of interior colonizers studied at the same time, those of surface colonizers observed on the fallen needles at four different times are roughly similar to each other.Thysanophora penicillioides was the major first colonizer on the sample needles from the O horizon, andTrichoderma species followed it. In an experiment started in late autumn, three dematiaceous fungi,Chloridium viride var.chlamydosporis, Sporidesmium omahutaense, andChalara sp., commonly occurred and contributed to the darkening of colonized needles. Seasonal variation in climate may have a stronger effect on internal colonizers than external colonizers of needles. Contributions from Sugadaira Montane Research Center, No. 165.     

Fungi on Leaf Blades of <Emphasis Type="Italic">Phragmites australis</Emphasis> in a Brackish Tidal Marsh: Diversity,Succession, and Leaf Decomposition

Although fungi are known to colonize and decompose plant tissues in various environments, there is scanty information on fungal communities on wetland plants, their relation to microhabitat conditions, and their link to plant litter decomposition. We examined fungal diversity and succession on Phragmites australis leaves both attached to standing shoots and decaying in the litter layer of a brackish tidal marsh. Additionally, we followed changes in fungal biomass (ergosterol), leaf nitrogen dynamics, and litter mass loss on the sediment surface of the marsh. Thirty-five fungal taxa were recorded by direct observation of sporulation structures. Detrended correspondence analysis and cluster analysis revealed distinct communities of fungi sporulating in the three microhabitats examined (middle canopy, top canopy, and litter layer), and indicator species analysis identified a total of seven taxa characteristic of the identified subcommunities. High fungal biomass developed in decaying leaf blades attached to standing shoots, with a maximum ergosterol concentration of 548 ± 83 μg g^–1 ash-free dry mass (AFDM; mean ± SD). When dead leaves were incorporated in the litter layer on the marsh surface, fungi experienced a sharp decline in biomass (to 191 ± 60 μg ergosterol g^–1 AFDM) and in the number of sporulation structures. Following a lag phase, species not previously detected began to sporulate. Leaves placed in litter bags on the sediment surface lost 50% of their initial AFDM within 7 months (k = −0.0035 day^–1) and only 21% of the original AFDM was left after 11 months. Fungal biomass accounted for up to 34 ± 7% of the total N in dead leaf blades on standing shoots, but to only 10 ± 4% in the litter layer. These data suggest that fungi are instrumental in N retention and leaf mass loss during leaf senescence and early aerial decay. However, during decomposition on the marsh surface, the importance of living fungal mass appears to diminish, particularly in N retention, although a significant fraction of total detrital N may remain associated with dead hyphae.     

The Role of Early Fungal Colonizers in Leaf‐Litter Decomposition in Portuguese Streams Impacted by Agricultural Runoff

We conducted a transplant experiment between two streams in NW Portugal impacted by agricultural runoff, mainly differing in phosphate concentration, to determine whether fungi on decomposing leaves would adapt to the new environment or would be replaced by fungi of the recipient stream. The most nutrient enriched stream had lower fungal diversity but faster leaf decomposition. Leaf transplantation did not alter fungal activity or species dominance. Multidimensional scaling ordination of fungal communities, from DNA fingerprint or conidial production, revealed that transplanted communities resembled more those of the original stream than the recipient stream. Results suggest that early fungal colonizers will determine the development and activity of fungal communities on decomposing leaves in streams impacted by agricultural practices. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Fungal Propagules and DNA in Feces of Two Detritus-Feeding Amphipods

Aquatic shredders (leaf-eating invertebrates) preferentially ingest and digest leaves colonized by aquatic hyphomycetes (fungi). This activity destroys leaf-associated fungal biomass and detritial resources in streams. Fungal counter-adaptations may include the ability to survive passage through the invertebrate's digestive tract. When fecal pellets of Gammarus tigrinus and Hyalella azteca were incubated with sterile leaves, spores of nine (G. tigrinus) and seven (H. azteca) aquatic hyphomycete species were subsequently released from the leaves, indicating the presence of viable fungal structures in the feces. Extraction, amplification, and sequencing of DNA from feces revealed numerous fungal phylotypes, two of which could be assigned unequivocally to an aquatic hyphomycete. The estimated contributions of major fungal groups varied depending on whether 18S or ITS sequences were amplified and cloned. We conclude that a variable proportion of fungal DNA in the feces of detritivores may originate from aquatic hyphomycetes. Amplified DNA may be associated with metabolically active, dormant, or dead fungal cells.     

Leaf growth,senescence and decomposition of Juncus maritimus Lam. in a coastal Mediterranean marsh

In this study, the growth, senescence, leaf loss and nutrient dynamics of Juncus maritimus were followed to examine litter decay in a Mediterranean coastal marsh. Decomposition was studied in dead leaves still attached to the plant and in leaves placed in litterbags (detached leaves/litter) on the sediment surface. The dynamics of fungi, meiofauna and epiphytes associated with detached litter were also followed. No significant differences were observed between decay rates in dead leaves attached to plants (0.0017 day⁻¹) and detached leaves (0.0015 day⁻¹) in litter bags. The percentage of ash-free dry weight lost was inversely proportional to the C:N and C:P ratios in plant detritus during decay, indicating N and P limitation for the decomposer community inhabiting decaying J. maritimus litter and uptake of these nutrients from the environment. Water availability and high temperatures on the sediment surface increased the density of meiofauna and epiphyton and decreased fungal biomass during the first 20 days of the experiment. The density of ciliates and nematodes in decomposing litter was inversely related to the C:N ratio and directly related to the percentage of AFDW lost. On the basis of these observations, it was concluded that meiofauna are primary colonizers of J. maritimus leaf litter.     

Leaf growth,senescence and decomposition of Juncus maritimus Lam. in a coastal Mediterranean marsh

In this study, the growth, senescence, leaf loss and nutrient dynamics of Juncus maritimus were followed to examine litter decay in a Mediterranean coastal marsh. Decomposition was studied in dead leaves still attached to the plant and in leaves placed in litterbags (detached leaves/litter) on the sediment surface. The dynamics of fungi, meiofauna and epiphytes associated with detached litter were also followed. No significant differences were observed between decay rates in dead leaves attached to plants (0.0017 day⁻¹) and detached leaves (0.0015 day⁻¹) in litter bags. The percentage of ash-free dry weight lost was inversely proportional to the C:N and C:P ratios in plant detritus during decay, indicating N and P limitation for the decomposer community inhabiting decaying J. maritimus litter and uptake of these nutrients from the environment. Water availability and high temperatures on the sediment surface increased the density of meiofauna and epiphyton and decreased fungal biomass during the first 20 days of the experiment. The density of ciliates and nematodes in decomposing litter was inversely related to the C:N ratio and directly related to the percentage of AFDW lost. On the basis of these observations, it was concluded that meiofauna are primary colonizers of J. maritimus leaf litter.     

Succession of microfungi during Pistacia lentiscus litter decomposition in a Sardinian Mediterranean maquis

Abstract

Changes in the microfungal community developing on Pistacia lentiscus L. leaf litter were monitored from February 2001 (senescent leaves) to December 2002 using litter bags. The Principal component analysis of the 16 samples showed a microfungal succession related to the progressive decomposition of the substratum. The Correspondence analysis of fungal colonizers with Rf > 10 revealed three main groups succeeding during the study period. The co-occurrence relationships (on the same leaf) among the major colonizers were analysed. The specialized saprotrophs on P. lentiscus litter: Asterostomella sp. 1 and Endophragmiella boewei were identified as principal colonizers in the mature community.     

Some observations on fungal succession during decomposition of wool in soil

Fungal succession on woolen baits was studied under laboratory conditions for more than one year. It was found that the initial colonizers on woolen baits are non-keratinophilic fungi, while the late colonizers are keratinophilic fungi. Six phases in total were observed during fungal succession. The successional trends obtained during decomposition of wool in soil samples collected from plain and hilly areas were almost the same, except for the dominant colonization in the last phase, which was constituted byChrysosporium tropicum for the plain, butMicrosporum gypseum andM. fulvum for the hilly area.     

Aquatic hyphomycetes in a changing environment

In 1942, Ingold documented an ecologically defined group of fungi, aquatic hyphomycetes, on autumn-shed leaves decaying in streams. They were shown to be vital intermediaries between the nutritionally poor leaf substratum and leaf-eating invertebrates. Research has subsequently emphasized functional aspects such as leaf decomposition and nutritional conditioning by fungi. Structural aspects (community composition) have attracted less attention, partly because of the difficulties of identifying fungal mycelia in situ. Extraction, amplification (PCR, qPCR) and characterization of DNA and RNA, and, more recently, of proteins, allow much greater insights into the presence of fungal taxa, their metabolic status (dead, dormant or active), and their potential and actual participation in decomposition processes. This approach can yield huge amounts of data, and major challenges today are the development and application of suitable bioinformatics techniques. The complexity of data collection and evaluation favour interdisciplinary teams of researchers. Fungi are major players in most ecosystems and are increasingly affected by human impacts. Changing land use, eutrophication/pollution and climate change are among the major factors that affect diversity and ecological functions of aquatic hyphomycetes.     

Influence of fungal-bacterial interactions on bacterial conjugation in the residuesphere

Conjugal gene transfer among bacteria in the residuesphere (area between decaying plant material and soil) of leaves of barley straw was studied. The residuesphere was shown to be a hot-spot for conjugal gene transfer compared to conjugation in sterile sand and non-sterile bulk soil. Impact of fungal colonisation of the residuesphere on bacterial colonisation and conjugation was also investigated. The inhibition of fungal colonisation, due to the application of an eukaryotic inhibitor, increased bacterial colonisation of the residuesphere in soil microcosms compared to non-treated leaves. This treatment also had a transient, positive effect on conjugation. Bacterial conjugation in the residuesphere of leaves subjected to 17 days of fungal colonisation was significantly lower than in the residuesphere of non-colonised leaves. Fungal biomass, as measured by chitinase activity, was inversely related to the conjugation efficiency.     

Autumnal biomass and potential productivity of salt marsh fungi from 29 degrees to 43 degrees north latitude along the United States Atlantic Coast

It has been established that substantial amounts of fungal mass accumulate in standing decaying smooth cordgrass (Spartina alterniflora) marshes in the southeastern United States (e.g., in standing decaying leaf blades with a total fungal organic mass that accounts for about 20% of the decay system organic mass), but it has been hypothesized that in marshes farther north this is not true. We obtained samples of autumnal standing decaying smooth cordgrass from sites in Florida to Maine over a 3-year period. The variation in latitude could not explain any of the variation in the living fungal standing crop (as determined by ergosterol content) or in the instantaneous rates of fungal growth (as determined by acetate incorporation into ergosterol at a standard temperature, 20 degrees C), which led to the conclusion that the potential levels of fungal production per unit of naturally decaying grass are not different in northern and southern marshes. Twenty-one percent of the variation in the size of the living fungal standing crop could be explained by variation in the C/N ratio (the higher the C/N ratio the smaller the fungal crop), but the C/P ratio was not related to the size of the fungal crop. Instantaneous rates of fungal growth were negatively related to the size of the living fungal crop (r = -0.35), but these rates were not correlated with C/nutrient ratios. The same two predominant species of ascomycetes (one Phaeosphaeria species and one Mycosphaerella species) were found ejecting ascospores from standing decaying smooth cordgrass blades at all of the sites examined from Florida to Maine.     

Rapid and Pervasive Occupation of Fallen Mangrove Leaves by a Marine Zoosporic Fungus

Samples of leaves of red mangrove (Rhizophora mangle) were incubated on an agar medium selective for pythiaceous oomycetes. Leaves on trees above the water did not contain oomycetes. Marine oomycetes, principally Phytophthora vesicula, had colonized leaves within 2 h of leaf submergence, probably finding them by chemotaxis. The frequency of occurrence of P. vesicula in submerged leaves reached 100% within 30 h of submergence. By 43 h most, if not all, parts of leaves were occupied, and surface treatment with a biocide indicated that leaves were occupied internally. Frequencies of P. vesicula remained near 100% through about 2 weeks of submergence and then declined to about 60% in older (≥4 weeks) leaves. Leaves of white mangrove (Laguncularia racemosa) were also extensively occupied by P. vesicula after falling into the water column, but decaying leaves of turtlegrass (Thalassia testudinum) were not colonized by oomycetes. Ergosterol analysis indicated that the standing crop of living, non-oomycete (ergosterol-containing) fungal mass in submerged red-mangrove leaves did not rise above that which had been present in senescent leaves on the tree; decaying turtlegrass leaves had an ergosterol content that was only about 2% of the maximum concentration detected for red-mangrove leaves. These results suggest that oomycetes are the predominant mycelial eucaryotic saprotrophs of mangrove leaves that fall into the water column and that for turtlegrass leaves which live, die, and decompose under submerged conditions, mycelial eucaryotes make no substantial contribution to decomposition.     

Minimizing Ergosterol Loss during Preanalytical Handling and Shipping of Samples of Plant Litter

Common preliminary treatments of samples of decaying material can involve changes in water content (e.g., via storage in relatively dry air or rinsing) that could conceivably result in loss or gain of fungal membranes and, consequently, ergosterol. A related problem is that collecting of ergosterol content data from widely distributed locales by shipment of samples ideally requires an inexpensive, safe alternative to submerging the samples in methanol for prevention of ergosterol loss. Experimental testing showed that fungal occupants of decaying salt marsh grass leaves did not exhibit loss or gain of ergosterol during air drying (to a water potential of <-8 MPa) or rewetting (to -0.8 MPa). Wet leaves of one grass species (Juncus roemerianus, black needlerush) could be fixed and dried for shipment by microwaving, or by fully drying after alcoholic or pentane fixation, without ergosterol loss, but those of smooth cordgrass (Spartina alterniflora) lost about 40% of their ergosterol content by all three of these drying methods. Ergosterol content of wet leaves of cordgrass could be maintained by alcoholic fixation and subsequent drying down to a thin film of alcohol.     

Autumnal Biomass and Potential Productivity of Salt Marsh Fungi from 29° to 43° North Latitude along the United States Atlantic Coast

It has been established that substantial amounts of fungal mass accumulate in standing decaying smooth cordgrass (Spartina alterniflora) marshes in the southeastern United States (e.g., in standing decaying leaf blades with a total fungal organic mass that accounts for about 20% of the decay system organic mass), but it has been hypothesized that in marshes farther north this is not true. We obtained samples of autumnal standing decaying smooth cordgrass from sites in Florida to Maine over a 3-year period. The variation in latitude could not explain any of the variation in the living fungal standing crop (as determined by ergosterol content) or in the instantaneous rates of fungal growth (as determined by acetate incorporation into ergosterol at a standard temperature, 20°C), which led to the conclusion that the potential levels of fungal production per unit of naturally decaying grass are not different in northern and southern marshes. Twenty-one percent of the variation in the size of the living fungal standing crop could be explained by variation in the C/N ratio (the higher the C/N ratio the smaller the fungal crop), but the C/P ratio was not related to the size of the fungal crop. Instantaneous rates of fungal growth were negatively related to the size of the living fungal crop (r = −0.35), but these rates were not correlated with C/nutrient ratios. The same two predominant species of ascomycetes (one Phaeosphaeria species and one Mycosphaerella species) were found ejecting ascospores from standing decaying smooth cordgrass blades at all of the sites examined from Florida to Maine.     

Productivities of microbial decomposers during early stages of decomposition of leaves of a freshwater sedge

1. We examined standing-senescing, standing-dead and recently fallen leaf blades of Carex walteriana in fens of the Okefenokee Swamp to determine the nature of the microbial decomposers in the early stages of decomposition, measuring both standing crops and productivities ([³H]leucineprotein method for bacteria, [¹⁴C]acetateergosterol for fungi). 2. Fungal standing crops (ergosterol) became detectable at the mid-senescence stage (leaves about half yellow-brown) and rose to 14–31 mg living-fungal C g^?1 organic mass of the decaying system; bacterial standing crops (direct microscopy) were ± 0.2 mgC g^?1 until the fallen-leaf stage, when they rose to as high as 0.9 mgC g^?1. 3. Potential microbial specific growth rates were similar between fungi and bacteria, at about 0.03–0.06 day^?1, but potential production of fungal mass was 115–512 μgC g^?1 organic mass day^?1, compared with 0–22 μgC g^?1 day^?1 for bacteria. Rates of fungal production were about 6-fold lower on average than previously found for a saltmarsh grass, perhaps because much lower phosphorus concentratiofis in the freshwater fen limit fungal activity. 4. There was little change in lignocellulose (LC) percentage of decaying leaves, although net loss of organic mass at the fallen, broken stage was estimated to be 59%, suggesting that LC was lost at rates proportional to those for total organics during decay. Monomers of fungal-wall polymers (glucosamine and mannose) accumulated 2- to 4-fold during leaf decay. This may indicate that an increase found for proximate (acid-detergent) lignin could be at least partially due to accumulation of refractory fungal-wall material, including melanin. 5. A common sequence in decaying aquatic grasses is suggested: principally fungal alteration of LC during standing decay, followed by a trend toward bacterial decomposition of the LC after leaves fall and break into particles.