Interrelationships between yeast fungi and collembolans in soil

The possibility of feeding on green and newly fallen leaves of the small-leaved lime Tilia cordata was studied for the collembolans Protaphorura armata and Vertagopus pseudocinereus. Young leaves grown under sterile conditions and almost free of yeast fungi were found to be toxic to the collembolan V. pseudocinereus: feeding on them led to the death of the animals. Leaves grown under natural conditions were nontoxic: when used by the collembolans as feed, they provided for collembolan growth and fecundity. Feeding preferences of the collembolans in relation to the yeasts attributed to different ecomorphs—epiphytes, litter saprophytes, pedobionts, and saccharobionts—were studied. Of the 24 yeast strains isolated from plant green parts, litter, and soil and assigned to eight species, no strain was revealed that was not used by the collembolans. However, certain yeast strains were preferable for the collembolans. The population of the V. pseudocinereus collembolans feeding on the yeast Rhodotorula glutinis (nss 31–4) exceeded that grown on Cryptococcus terricola (2044) 1.5-fold. Thus, the collembolans have feeding preferences in relation to yeast fungi, as was shown earlier with mycelial micromycetes. The possible mechanisms of the feeding preferences of the collembolans in relation to yeasts are discussed.     

Selective foraging of fungi by collembolans in soil

Soils contain highly diverse communities of microorganisms and invertebrates. The trophic interactions between these species are largely unknown. Collembolans form an abundant part of the invertebrate community in soils. A prevailing view is that soil collembolans are generalist feeders on fungi, lichens, fragmented litter and bacteria. However, in laboratory food choice experiments, it has been shown that collembolans preferentially select certain taxa of fungi. To examine this apparent contradiction, we developed a molecular technique based on the analysis of 18S ribosomal DNA (rDNA) sequences to explore the diversity of fungi in soils and in the guts of collembolans. We report that the diversity of fungi found in the natural soil was 33 times higher than that in the guts of the collembolan Protaphorura armata. The data support the view that collembolan species can be highly selective when foraging on fungi in soils.     

Reproductive significance of feeding on saprobic and arbuscular mycorrhizal fungi by the collembolan, Folsomia candida

1. Collembolans have often been credited with negatively affecting arbuscular mycorrhizal (AM) symbioses, mainly by grazing and severing the associated external fungal network from host roots. However, most previous experiments were performed using relatively 'clean' systems where other, non-mycorrhizal, fungi were largely excluded. Yet, plant rhizospheres harbour a wide variety of highly palatable non-AM fungi, most of which have saprobic lifestyles.
2. In this study we isolated and cultured several rhizosphere fungi, and the collembolan , Folsomia candida , from the Long-Term Mycorrhiza Research Site, University of Guelph, Canada, to test the hypothesis that, given a choice, collembolans would prefer to feed on saprobic fungi and that such a choice is of adaptive significance to the animals.
3. A laboratory food preference experiment revealed that F. candida favours common saprobic fungi over a variety of AM fungi. Coincidentally, fecundity levels across two Folsomia generations were higher when animals fed exclusively on the preferred fungus, Alternaria alternata . When fed less palatable fungi, fecundity was greatly reduced; in fact animals from the F1 generation were unable to produce any eggs when placed on an exclusive diet of one of the following three AM fungi, Acaulospora spinosa, Scutellospora calospora and Gigaspora gigantea .
4. These results indicate that a strict diet of AM fungi by collembolans has reproductive consequences. Therefore, we propose that under natural conditions these animals spend more time feeding on common saprobic fungi rather than their AM counterparts. This suggests that previous 'clean' studies that investigated the interactions between collembolans and AM fungi may have reported exaggerated effects of animal grazing. The influence of collembolans on the functioning of AM symbioses, under more natural conditions, remains not well understood.     

Large 13C/12C and small 15N/14N isotope fractionation in an experimental detrital foodweb (litter–fungi–collembolans)

Correctly estimating the trophic fractionation factors (Δ¹⁵N and Δ¹³C) in controlled laboratory conditions is essential for the application of stable isotope analysis in studies on the trophic structure of soil communities. Laboratory experiments usually suggest large ¹⁵N/¹⁴N and small ¹³C/¹²C trophic fractionation, but in field studies litter-dwelling microarthropods and other invertebrates are consistently enriched in ¹³C relative to plant litter. In the present study, we report data from two laboratory experiments investigating both fungi–collembolans and litter–fungi–collembolans systems. In the fungi–collembolans system, Δ¹⁵N and Δ¹³C averaged 1.4 ± 0.1 and 1.0 ± 0.2 ‰, respectively. In microcosms with fungi-inoculated litter, the difference in δ¹⁵N between collembolans and plant litter averaged 1.5 ± 0.2 ‰, confirming the relatively small ¹⁵N/¹⁴N trophic fractionation at the basal level of detrital foodwebs reported in numerous field studies. In full agreement with field observations, the difference in δ¹³C between bulk litter and collembolans in laboratory microcosms averaged 3.6 ± 0.1 ‰ and only little depended on collembolan species identities or the presence of water-soluble compounds in the litter. We conclude that increased δ¹³C values typical of litter-dwelling decomposers are largely determined by an increased ¹³C content in saprotrophic microorganisms.     

Consequences of the colonisation of leaves by fungi and oomycetes for leaf consumption by a gammarid shredder

1. Leaf litter breakdown by shredders in the field is affected by leaf toughness, nutritional value and the presence of secondary compounds such as polyphenols. However, experiments involving the use of single fungal strains have not supported the assumption that leaf parameters determine food selection by shredders perhaps because of a failure to test for high consumption prior to isolation of fungal strains, overrepresentation of hyphomycetes or the potential effects of accompanying bacteria. In this study, we used bacteria‐free, actively growing fungi and oomycetes isolated from conditioned leaf litter for which a shredder had already shown high consumption rates. 2. Black alder (Alnus glutinosa) leaf litter was exposed to the littoral zone of Lake Constance in autumn, and subsamples were analysed for leaf parameters and consumption by Gammarus roeselii under standard conditions at regular intervals. On dates with a high consumption rate of the exposed leaves, 14 single strains of fungi and oomycetes were isolated, freed of bacteria and grown on autoclaved leaves. 3. Six of eight measured leaf parameters of exposed leaves were significantly correlated with Gammarus consumption rates, with high colinearity among leaf parameters hampering the identification of causal relations between leaf parameters and feeding activity. 4. When single strains of fungi and oomycetes were grown on autoclaved leaf litter, toughness of colonised leaves was always lower than in the control and the content of protein, N and P were increased. There were pronounced strain‐specific effects on leaf parameters. Consumption rates also differed significantly, with nine of fourteen isolates consumed at higher rates than controls and none proving to be a deterrent. Protein and polyphenol content were significantly correlated with consumption rates. Oomycete‐colonised leaves were consumed at similar rates but were of lower food quality than fungi‐colonised leaves. 5. We argue that direct strain‐specific attractant or repellent effects of fungi and oomycetes on consumption by G. roeselii are not important. However, we found indirect strain‐specific role operating via effects on leaf parameters.     

Influence of aeration during propagation of pitching yeast on fermentation and beer flavor

The effect of yeast propagated at different aeration conditions on yeast physiology, fermentation ability, and beer quality was investigated using three strains of Saccharomyces cerevisiae. It was shown that yeast cells grown under continuous aeration conditions during propagation were almost two times higher as compared with discontinuous aeration conditions. The maximum of cell growth of all samples reached between 36 h and 48 h. The concentration of trehalose was increased under continuous aerated yeasts, whereas glycogen was decreased. It was also observed that the concentration of glycogen and trehalose in yeast cells had no direct effect on subsequent fermentation ability. The effect of yeast propagated under different aeration conditions on subsequent fermentation ability was different from yeast strains, in which the influence will be most pronounced at the first fermentation. Later, the yeasts might regain its original characteristics in the following fermentations. Generally, continuously propagated yeast had a positive effect on beer quality in subsequent fermentation. Hence, the concentration of aroma compounds obtained with yeast propagated under 6 1/h for 48 h aeration was lower than those grown under other aeration conditions in the bottom yeasts; in particular, the amounts of phenylethyl alcohol, ester, and fatty acids were decreased.     

Effects of fungal food quality and starvation on the fatty acid composition of Protaphorura fimata (Collembola)

The lipid pattern of animals is influenced by species, life stage, environmental conditions and diet. We investigated the effects of food quality and starvation on the phospholipid (PLFA) and neutral lipid (NLFA) fatty acid pattern of the collembolan Protaphorura fimata. Collembolans were fed with two common soil fungi, Agrocybe gibberosa and Chaetomium globosum, of which the cellular lipid composition was analysed. A. gibberosa was grown on agar with different nitrogen contents, resulting in altered fatty acid patterns and C:N ratios, i.e. fungi of different food quality. Collembolans did not mirror the lipid composition of the fungal diet as the pattern of major NLFAs in P. fimata was vice versa. Presumably, altered food quality of fungi caused compensatory responses by the collembolans, thereby diminishing the fungal signal. In a further experiment P. fimata (previously maintained with C. globosum) was kept without food for up to 4 weeks. Starvation resulted in a decline in the total amount of NLFAs; however, it did not affect the fatty acid pattern, indicating that NLFAs were degraded indiscriminately. Generally, the PLFA profile of the collembolans changed only slightly due to variations in diet quality or starvation.     

Zoophagy in Collembola

The feeding of collembolans on animal food, including nematodes and small enchytraeids, was studied in laboratory. Active preying on worms was found in more than 30 Collembola species. Fecundity and natural growth of the collembolan populations were similar in the case of separate diet (fungal and animal), but mixed feeding increased both parameters by several times. No special hunting behavior was found, and feeding on nematodes was very similar to gathering. Several collembolans attack small enchytraeids together, dismembering living worms.     

Patchiness and compensatory growth in a fungus-Collembola system

The compensatory growth potential of a grazed fungal biomass was mathematically expressed as a function of patchiness in its distribution and demonstrated in an experiment using the fungivorous collembolan Onychiurus armatus and the soil fungi Verticillium bulbillosum and Penicillium spinulosum. The model addresses the regrowth potential in relation to patch fragmentation, travelling time and consumption rate of the collembolan and the mean relative growth rate of the fungus. It suggests that the mean relative growth rate required for regrowth decreases with patch fragmentation and increases with the mean growth rate of the fungus. The experiments were performed with a system of soil-filled vials provided with fungi and collembolans. The size of the vials and the length of the tubes connecting them were varied to give different patch sizes and travelling times. The respiratory activity of fungi after grazing increased as a unit of mycelium was distributed into smaller connected vials. The slow growing species V. bulbillosum showed a greater but delayed response to grazing in comparison with the fast growing P. spinulosum. An increased travelling time delayed the growth response in both species.     

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.     

Quantitative and qualitative seasonal changes in the microbial community from the phyllosphere of sugar beet (Beta vulgaris)

Bacteria, yeasts and filamentous fungi colonizing immature, mature and senescing primary leaves of field grown Beta vulgaris (sugar beet) were analysed over a complete growing season. Greatest microbial numbers were detected on senescing primary leaves and these numbers increased over most of the season. The number of colonizers detected on mature leaves was found to be stable over most of the study.Filamentous fungi and yeasts were identified to the genus level and the communities found to have greatest diversity during the summer months. There was no consistent pattern of diversity according to leaf type. Two genera of filamentous fungi, Cladosporium and Alternaria and two yeast genera, Cryptococcus and Sporobolomyces were the most numerous fungal populations isolated. Only 8 filamentous fungi and 3 yeast genera were commonly isolated on PDA (potato dextrose agar).Bacterial strains (1236) were isolated on Tryptic Soy Broth (TSB) agar and identified to species, or in some cases sub-species level, by analysis of their fatty acid methyl ester (FAME) profiles. Isolated bacteria were grouped into 78 named and 37 unnamed species clusters. Greatest number of bacterial species were isolated from young plants and leaves, sampled during the autumn months. Bacterial community diversity was lowest in mid-summer and winter months. Pseudomonas was the most commonly isolated genus and Erwinia herbicola the most common species. P. aureofaciens was the only species isolated from soil that was also isolated from the phyllosphere of B. vulgaris throughout the season.     

Repellency of injured ascomata of Ciborinia camelliae and Spathularia flavida to fungivorous collembolans

The repellency of injured ascomata of Ciborinia camelliae and Spathularia flavida was tested with the collembolan Ceratophysella denisana, a common mushroom feeder. Presentation of a test ascoma (intact or injured) at a distance to a collembolan that was feeding at a bait showed that significantly more collembolans moved away in response to the presentation of injured ascomata than intact ones for both species. These results suggest that the ascomata of these species showed repellency to the collembolan species as a response to injury. Odor is speculated to be one of the stimuli eliciting the behavioral responses observed.     

Barley pathogenesis-related proteins with fungal cell wall lytic activity inhibit the growth of yeasts.

Proteins from intercellular fluid extracts of chemically stressed barley (Hordeum vulgare L.) leaves were separated by native polyacrylamide gel electrophoresis at alkaline or acid pH. Polyacrylamide gels contained Saccharomyces cerevisiae (bakers' yeast) or Schizosaccharomyces pombe (fission yeast) crude cell walls for assaying yeast wall lysis. In parallel, gels were overlaid with a suspension of yeasts for assaying growth inhibition by pathogenesis-related proteins. The same assays were also performed with proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions. In alkaline native polyacrylamide gels, only one band corresponding to yeast cell wall lytic activity was found to be inhibitory to bakers' yeast growth, whereas in acidic native polyacrylamide gels one band inhibited the growth of both yeasts. Under denaturing nonreducing conditions, one band of 19 kD inhibited the growth of both fungi. The 19-kD band corresponded to a basic protein after two-dimensional gel analysis. The 19-kD protein with yeast cell wall lytic activity and inhibitory to both yeasts was found to be different from previously reported barley chitosanases that were lytic to fungal spores. It could be different from other previously reported lytic antifungal activities related to pathogenesis-related proteins.     

Revisiting the phylogeny of Wolbachia in Collembola

The endosymbiont Wolbachia has been detected in a few parthenogenetic collembolans sampled in Europe and America, including three species of Poduromorpha, two species of Entomobryomorpha, and two species of Neelipleona. Based on 16S rRNA and ftsZ gene sequences, most of the Wolbachia infecting parthenogenetic collembolans were characterized as members of supergroup E and showed concordant phylogeny with their hosts. However, the two neelipleonan symbionts form another unique group, indicating that Wolbachia has infected parthenogenetic collembolans multiple times. In this study, five parthenogenetic collembolan species were identified as hosts of Wolbachia, and four new Wolbachia strains were reported for four collembolan species sampled in China, respectively, including a neelipleonan strain from Megalothorax incertus (wMinc). Our results demonstrated that the Wolbachia multilocus sequence typing (MLST) system is superior to the 16S rRNA + ftsZ approach for phylogenetic analyses of collembolan Wolbachia. The MLST system assigned these Wolbachia of parthenogenetic collembolans to supergroup E as a unique clade, which included wMinc, supporting the monophyletic origin of Wolbachia in parthenogenetic collembolan species. Moreover, our data suggested supergroup E as one of the most divergent lineages in Wolbachia and revealed the discrepancy between the phylogenies of Wolbachia from parthenogenetic collembolans and their hosts, which may result from the high level of genetic divergence between collembolan Wolbachia, in association with the geographic differentiation of their hosts, or the possible horizontal transmission of Wolbachia between different collembolan species.     

Effects of elevated carbon dioxide on green leaf tissue and leaf litter quality in an intact Mojave Desert ecosystem

The effects of elevated carbon dioxide (CO₂) on plant litter are critical determinants of ecosystem feedback to changing atmospheric CO₂ concentrations. We measured concentrations of nitrogen (N) and carbon (C) and calculated C : N ratios of green leaves of two desert perennial shrubs, and the same quality parameters plus lignin and cellulose content of leaf litter from four shrub species exposed to elevated CO₂ (FACE technology; Hendrey & Kimball, 1994 ) for 3 years in an intact Mojave Desert ecosystem. Shrubs tested were Larrea tridentata, Lycium pallidum, Lycium andersonii and Ambrosia dumosa. We calculated resorption efficiency from green tissue and leaf litter N data and measured lignin and cellulose content in litter in the last year study. Green leaves of L. tridentata grown under elevated CO₂ had significantly lower N concentrations and higher C : N ratios than shrubs grown in ambient conditions in 1999 (P < 0.05). Lycium pallidum green leaves grown under elevated CO₂ had significantly lower N concentrations and higher C : N ratios than shrubs grown under ambient conditions in 2000 (P < 0.05). There was no CO₂ effect on C content of either species. We found no effect of CO₂ on N or C content, C : N ratios, or lignin or cellulose concentrations in leaf litter of L. tridentata, L. pallidum, L. andersonii, or A. dumosa. There was no significant effect of CO₂ on estimates of shrub resorption efficiency. There was a seasonal effect on green tissue and litter tissue quality for L. tridentata, with lower tissue N content in summer than in spring or winter months. These data suggest that any productivity increases with elevated CO₂ in desert ecosystems may not be limited by lower leaf litter quality and that resorption efficiency calculations are best performed on an individual leaf basis.     

Feeding plasticity of two detritivore-shredders

1. The feeding preferences of the trichopteran shredder Sericostoma personatum Spence and the amphipod shredder Gammarus pulex L. were studied using specimens collected in a springbrook where the major food source was beech litter (Fagus sylvatica L.). Six food items were tested: conditioned beech leaves, conditioned alder leaves (Alnus glutinosa L.), conditioned Sitka spruce needles [Picea sitchensis (Bong.) Carr], fresh beech leaves, a fresh macrophyte (Potamogeton perfoliatus L.) and a fresh filamentous green alga (Microspora sp.). 2. The overall preference pattern shown by the two shredders was the same: both preferred conditioned Alnus most, followed by fresh Microspora. The least preferred food items were conditioned Fagus leaves and Picea needles. 3. The feeding preference of the two shredders proved to be unrelated to food source fibre content, toughness, total phosphorus, C:N ratio and total nitrogen (P>0.05). 4. Despite the overall similarity in their feeding preferences, Gammarus was more selective than Sericostoma. The reason for this difference is discussed. 5. We interpret our findings as indicating that detritivore-shredders do not per se prefer leaf litter, but in fact actively select other food items such as filamentous green algae or macrophytes, even when terrestrial leaf litter is abundant. Most shredders in Danish forest streams thus seem to live on a growth-limiting food resource that they do not prefer. This may have important implications for secondary production in such streams.     

Transcriptional regulation of fermentative and respiratory metabolism in Saccharomyces cerevisiae industrial bakers' strains

Bakers' yeast-producing companies grow cells under respiratory conditions, at a very high growth rate. Some desirable properties of bakers' yeast may be altered if fermentation rather than respiration occurs during biomass production. That is why differences in gene expression patterns that take place when industrial bakers' yeasts are grown under fermentative, rather than respiratory conditions, were examined. Macroarray analysis of V1 strain indicated changes in gene expression similar to those already described in laboratory Saccharomyces cerevisiae strains: repression of most genes related to respiration and oxidative metabolism and derepression of genes related to ribosome biogenesis and stress resistance in fermentation. Under respiratory conditions, genes related to the glyoxylate and Krebs cycles, respiration, gluconeogenesis, and energy production are activated. DOG21 strain, a partly catabolite-derepressed mutant derived from V1, displayed gene expression patterns quite similar to those of V1, although lower levels of gene expression and changes in fewer number of genes as compared to V1 were both detected in all cases. However, under fermentative conditions, DOG21 mutant significantly increased the expression of SNF1 -controlled genes and other genes involved in stress resistance, whereas the expression of the HXK2 gene, involved in catabolite repression, was considerably reduced, according to the pleiotropic stress-resistant phenotype of this mutant. These results also seemed to suggest that stress-resistant genes control desirable bakers' yeast qualities.     

Le Paraquat, un Outil pour la Révélation Rapide d'Infections Fongiques Latentes et de Champignons Endophytes

Paraquat: a tool for the quick development of latent fungal infections and endophytic fungi.
Detached winter wheat leaves without any symptom of fungal infection were thoroughly washed, surface desinfected, treated with paraquat (0.03–0.32% a.i.) and incubated on nutrient agar or in moist chambers under sterile conditions. Microscopic examinations showed that hypae and yeast cells emerged from several stomata within 48–72 h after treatment. Fungal colonies developed on agar within the same period of time. More than 27 filamentous fungal genera and yeasts were identified during a wheat cropin healthy, green leaves. Non-treated leaves showed very few early emergences of fungi from stomata, but senescent leaves eventually yielded several fungal taxa observed after paraquat treatment. Similar observations were made on non-detached wheat leaves, and on detached sugarbeet leaves. Paraquat also induced development of Fusarium spp. from detached healthy winter wheat ears before or after heading. In grape berries, paraquat induced the development of latent infections of Botrytis cinerea , a few days after treatment.
The usefulness of paraquat for the detection of endophytes and for the diagnosis of early and latent fungal infections in plants, and the possible involvement of endophytes in tissue senescence are discussed.     

CARDIOLIPIN CONTENT OF WILD TYPE AND MUTANT YEASTS IN RELATION TO MITOCHONDRIAL FUNCTION AND DEVELOPMENT

The phospholipid composition of various strains of the yeast, Saccharomyces cerevisiae, and several of their derived mitochondrial mutants grown under conditions designed to induce variations in the complement of mitochondrial membranes has been examined. Wild type and petite (cytoplasmic respiratory deficient) yeasts were fractionated into various subcellular fractions, which were monitored by electron microscopy and analyzed for cytochrome oxidase (in wild type) and phospholipid composition. 90% or more of the phospholipid, cardiolipin was found in the mitochondrial membranes of wild type and petite yeast. Cardiolipin content differed markedly under various growth conditions. Stationary yeast grown in glucose had better developed mitochondria and more cardiolipin than repressed log phase yeast. Aerobic yeast contained more cardiolipin than anaerobic yeast. Respiration-deficient cytoplasmic mitochondrial mutants, both suppressive and neutral, contained less cardiolipin than corresponding wild types. A chromosomal mutant lacking respiratory function had normal cardiolipin content. Log phase cells grown in galactose and lactate, which do not readily repress the development of mitochondrial membranes, contained as much cardiolipin as stationary phase cells grown in glucose. Cytoplasmic mitochondrial mutants respond to changes in the glucose concentration of the growth medium by variations in their cardiolipin content in the same way as wild type yeast does under similar growth conditions. It is concluded that cardiolipin content of yeast is correlated with, and is a good indicator of, the state of development of mitochondrial membrane.     

Microbial introductions to apple leaves: Influences of altered immigration on fungal community dynamics

Fungal immigration to apple leaves in the field was altered by the introduction of populations ofChaetomium globosum orAureobasidium pullulans to surface-disinfested leaves either immediately following, or 6 days after, disinfestation. Total numbers of fungal individuals and numbers of filamentous fungal and yeast individuals were estimated and compared over time for 4–7 weeks on control leaves (leaves disinfested but no populations applied), onAureobasidium-treated, and onChaetomium-treated leaves. Fungal communities developing on leaves during three experiments in two different time frames (experiment 1: July 9–August 27; experiments 2 and 3: July 29–August 27), and thus under different immigration regimes, were also compared. Survival of introduced populations was not related to the presence of prior fungal immigrants. Rates of increase in total numbers of fungi and numbers of filamentous fungi and yeasts per leaf varied among experiments, apparently in relation to differences in immigration and environmental history. Differences among leaves in immigration had a short-term (days) influence on community size. However, no long-term effects of altered immigration on phylloplane fungal community size were evident.