Preliminary insights into the evolutionary relationships of aquatic hyphomycetes and endophytic fungi

Aquatic hyphomycetes play a key role in leaf litter decomposition and are mediators of organic matter turnover in streams. Molecular studies have shown that some aquatic fungi are also plant endophytes, however, more evidence is needed to evaluate their multiple ecological abilities. To date, little information is available on fungal lineages that might have undergone convergent evolution to adapt to multiple ecological modes. We examined the phylogenetic relationships and evolutionary divergences of aquatic hyphomycetes, endophytic aquatic hyphomycetes and other fungal endophytes of riparian/terrestrial plants by analyzing ITS1-5.8S-ITS2 sequences retrieved from the National Center for Biotechnology Information (NCBI). Sequences with close phylogenetic affinity to aquatic fungi can occur as endophytes of terrestrial plants or in soil far from streams. To fully understand the ecological impact of aquatic hyphomycetes, we need to document and interpret their niches more broadly.     

Fungal communities on decaying leaves in streams: a comparison of two leaf species

Decomposition of leaf litter is a microbial mediated process that helps to transfer energy and nutrients from leaves to higher trophic levels in woodland streams. Generally, aquatic hyphomycetes are viewed as the major fungal group responsible for leaf litter decomposition. In this study, traditional microscopic examination (based on identification of released conidia) and phylogenetic analysis of 18S rRNA genes from cultivated fungi were used to compare fungal community composition on decomposing leaves of two species (sugar maple and white oak) from a NE Ohio stream. No significant differences were found in sporulation rates between maple and oak leaves and both had similar species diversity. From the 18S rRNA gene sequence data, identification was achieved for 12 isolates and taxonomic affiliation of 12 of the remaining 14 isolates could be obtained. A neighbor-joining tree (with bootstrap values) was constructed to examine the taxonomic distribution of the isolates relative to sequences of known operational taxonomic units (OTUs). Surprisingly, only 2 of the isolates obtained were aquatic hyphomycetes based on phylogenetic analysis. Overall, there were no differences between the two leaf types and a higher diversity was observed via culturing and subsequent 18S rRNA gene sequencing than by conidia staining. These differences resulted from the fact that traditional microscopy provides estimates of aquatic hyphomycete diversity while the other approach revealed the presence of both aquatic hyphomycete and non-aquatic hyphomycete taxa. The presence of this broad array of taxa suggests that the role of aquatic hyphomycetes relative to other fungi be re-evaluated. Even though the functional role of these non-aquatic hyphomycetes taxa is unknown, their presence and diversity demonstrates the need to delve further into fungal community structure on decomposing leaves.     

Decomposition of deciduous leaf litter in a woodland stream

Microorganisms associated with decomposing deciduous leaf litter in a woodland stream were examined by scanning electron microscopy. The use of a critical point drying method allowed the preservation of a wide variety of microorganisms as well as the decomposing litter with a minimum of distortion. The micrographs provide evidence that the aquatic hyphomycetes are the major fungal flora present during decomposition. Two distinct groups of these fungi were found during the seasonal cycle with one group occurring only in the summer while the other occurred throughout the rest of the year. The presence of all developmental stages of these organisms in the environment is considered further evidence of their active role in the decomposition of litter.     

Decomposition of deciduous leaf litter in a woodland stream

Microorganisms associated with decomposing deciduous leaf litter in a woodland stream were examined by scanning electron microscopy. The use of a critical point drying method allowed the preservation of a wide variety of microorganisms as well as the decomposing litter with a minimum of distortion. The micrographs provide evidence that the aquatic hyphomycetes are the major fungal flora present during decomposition. Two distinct groups of these fungi were found during the seasonal cycle with one group occurring only in the summer while the other occurred throughout the rest of the year. The presence of all developmental stages of these organisms in the environment is considered further evidence of their active role in the decomposition of litter.     

ADVANCES IN THE STUDY OF SO-CALLED AQUATIC HYPHOMYCETES

Progress in the study of the freshwater aquatic fungal flora (mainly hyphomycetes) of submerged decaying leaves of dicotyledonous trees and shrubs, since the discovery of the flora in 1942, is reviewed. The abundant occurrence of liberated conidia, which tend to be of distinctive form, in persistent foam on a stream is noted and illustrated by an example from Scotland. Developments in the taxonomy of these fungi since 1942 are noted. About 60 genera and around 120 species are now recorded. In a number the perfect stage has been discovered. In most cases it has proved to be an ascomycete, but it is sometimes a basidiomycete. Conidium development in aquatic hyphomycetes is discussed in the light of recent views on the classification of conidial types. It is argued that the most valid distinction is between phialoconidia and other types, rather than between those of blastic and those of thallic development. Dispersal is discussed with special reference to upstream spread and the problem of dispersal from one isolated water system to another. Finally, attention is given to the ecology and general physiology of these fungi. Their possible importance in processing decaying leaves for subsequent consumption by small aquatic animals is considered. The terrestrial occurrence of many “aquatic hyphomycetes” is noted and it is suggested that “amphibious hyphomycetes” might be a better term.     

Potential of aquatic fungi derived from diverse freshwater environments to decolourise synthetic azo and anthraquinone dyes

A total of 37 strains of aquatic hyphomycetes and 95 fungal isolates derived from diverse freshwater environments were screened on agar plates for the decolourisation of the disazo dye Reactive Black 5 and the anthraquinone dye Reactive Blue 19. The decolourisation of 9 azo and 3 anthraquinone dyes by 9 selected aquatic fungi was subsequently assessed in a liquid test system. The fungi were representatives of mitosporic anamorphs, and 6 strains had proven ascomycete affiliations. For comparison, 5 white rot basidiomycetes were included. The majority of dyes were decolourised by several mitosporic aquatic isolates at rates essentially comparable to those observed with the most efficient white rot fungus. Under certain conditions, particular aquatic strains decolourised dyes even more efficiently than the best performing white rot basidiomycete. Upon fungal treatment of several dyes, new absorbance peaks appeared, indicating biotransformation metabolites. All together, these results point to the potential of fungi occurring in freshwater environments for the treatment of dye-containing effluents.     

Lentic and lotic habitats as templets for fungal communities: traits,adaptations, and their significance to litter decomposition within freshwater ecosystems

Decomposition of plant matter is a key ecosystem process and considerable research has examined plant litter decay processes in freshwater habitats. Fungi are common inhabitants of the decomposer microbial community and representatives of all major fungal phyla have been identified within freshwater systems. Development and application of quantitative methods over the last several decades have firmly established that fungi are central players in the decomposition of plant litter in freshwaters and are important mediators of energy and nutrient transfer to higher trophic levels. Despite the critical roles that fungi play in carbon and nutrient cycling in freshwater ecosystems, there are notable differences in the types and adaptations of fungal communities between lotic and lentic habitats. These differences can be explained by the wide range of hydrologic, physical, chemical and biological conditions within freshwater systems, all of which can influence the presence, type, and activity of fungal decomposers and their impact on litter decomposition. This paper seeks to provide a brief overview of the types, adaptations, and role of fungi within lotic and lentic freshwater ecosystems, with a particular emphasis on their importance to litter decomposition and the key environmental conditions that impact their growth and decay activities. This discussion will specifically focus on fungal dynamics occurring on plant litter in forested headwater streams and emergent freshwater marshes, since published data concerning their role in these systems is considerably more abundant in comparison to other freshwater habitats.     

水生丝孢菌研究进展

本文回顾了自1942年水生丝孢菌发现以来,该类群的研究进展。世界范围内已有约80属、200种的水生丝孢菌被报道。中国发现20属、28种。12属中的15种水生丝孢菌的有性阶段已发现,它们大多数是子囊菌,少数为担子菌。生理学研究证实,该类真菌普遍地可以分泌胞外多糖酶,从而能较好地利用多种碳源。其它生理特性,如氮源和矿物的需求等,本文也作了综述。一些植物寄生性水生丝孢菌也有报道。生态学研究表明,秋季和冬季是水生丝孢菌大发生季节。它们在水环境中的重要作用,文中也作了较详细的讨论。最后涉及的是水生丝孢菌的传布和陆生情况的发现。本文还对该类群真菌的研究前景作了展望。     

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.     

Diversity,role in decomposition,and succession of zoosporic fungi and straminipiles on submerged decaying leaves in a woodland stream

Leaf litter is a very important primary source of energy in woodland streams. Decomposition of leaf litter is a process mediated by many groups of microorganisms which release extracellular enzymes for the degradation of complex macromolecules. In this process, true fungi and straminipiles are considered to be among the most active groups, more active than the bacteria, at least during the early stages of the process. Colonization increases the quality of the leaves as a food resource for detritivores. In this way, matter and energy enter detritus-based food chains. Previously, aquatic hyphomycetes were considered to be the major fungal group responsible for leaf litter decomposition. Although zoosporic fungi and straminipiles are known to colonize and decompose plant tissues in various environments, there is scant information on their roles in leaf decomposition. This study focuses on the communities of zoosporic fungi and straminipiles in a stream which are involved in the decomposition of leaves of two plant species, Ligustrum lucidum and Pouteria salicifolia, in the presence of other groups of fungi. A characteristic community dominated by Nowakowskiella elegans, Phytophthora sp., and Pythium sp. was found. Changes in the fungal community structure over time (succession) was observed: terrestrial mitosporic fungi appeared during the early stages, zoosporic fungi, straminipiles, and aquatic Hyphomycetes in early-to-intermediate stages, while representatives of the phylum Zygomycota were found at early and latest stages of the decomposition. These observations highlight the importance of zoosporic fungi and straminipiles in aquatic ecosystems.     

Leaf litter decomposition and microbial activity in nutrient-enriched and unaltered reaches of a headwater stream

SUMMARY 1. Decomposition of red maple ( Acer rubrum ) and rhododendron ( Rhododendron maximum ) leaves and activity of associated microorganisms were compared in two reaches of a headwater stream in Coweeta Hydrologic Laboratory, NC, U.S.A. The downstream reach was enriched with ammonium, nitrate, and phosphate whereas the upstream reach was not altered.
2. Decomposition rate, microbial respiration, fungal and bacterial biomass, and the sporulation rate of aquatic hyphomycetes associated with decomposing leaf material were significantly higher for both leaf types in the nutrient-enriched reach. Species richness and community structure of aquatic hyphomycetes also exhibited considerable changes with an increase in the number of fungal codominants in the nutrient-enriched reach.
3. Fungal biomass was one to two orders of magnitude greater than bacterial biomass in both reaches. Changes in microbial respiration rate corresponded to those in fungal biomass and sporulation, suggesting a primary role of fungi in leaf decomposition.
4. Nutrient enrichment increased microbial activity, the proportion of leaf carbon channelled through the microbial compartment and the decomposition rate of leaf litter.     

Lignicolous freshwater fungi along a north–south latitudinal gradient in the Asian/Australian region; can we predict the impact of global warming on biodiversity and function?

Fungi play a key role in decomposition of submerged wood in streams, breaking down lignocelluloses and releasing nutrients, and are important in ecosystem functioning. These wood decay fungi are known as freshwater lignicolous fungi and are usually studied by collecting submerged woody litter, followed by incubation in a moist chamber. This review explains what are freshwater lignicolous fungi, their decay mechanisms, roles and physiological attributes. Asian/Australasian lignicolous freshwater fungi have been relatively well-surveyed and enable an account of their distribution along a latitudinal transect. Unlike freshwater leaf-dwelling fungi their diversity in water bodies is greater towards the Equator which suggests they are important for decaying submerged wood in the tropics. Riparian vegetation, disturbances such as pollution, streams drying and study methods, may all affect the diversity of freshwater lignicolous fungi, however, the overall trend is a higher diversity in the tropics and subtropics. Climate changes together with increasing deposition of woody debris from human activities, and alteration of environmental factors (such as water pollution, and dam building) will impact freshwater lignicolous fungi. Changing diversity, structure and activities of freshwater fungal communities can be expected, which will significantly impact on aquatic ecosystems, particularly on nutrient and carbon cycles. There is a great opportunity to monitor changes in freshwater fungi communities along latitudinal (north to south) and habitat gradients (from human disturbed to natural habitats), and study ecological thresholds and consequences of such changes, particularly its feedback on nutrient and carbon cycles in freshwater systems.     

Biodiversity of freshwater fungi

There are more than 600 species of freshwater fungi with more known from temperate, as compared to tropical regions. These includeca 340 ascomycetes, 300 deuteromycetes, and a number of lower fungi which are not discussed here.Aniptodera, Annulatascus, Massarina, Ophioceras andPseudohalonectria are common freshwater ascomycetes, which appear to be well adapted for this lifestyle either in their ascospore types or their competitive-degradative characters. The most common genera of wood-inhabiting deuteromycetes includeCancellidium, Dactylaria, Dictyosporium andHelicomyces. They are categorized into four groups depending on their form and life style: the ingoldian hyphomycetes; the aero-aquatic hyphomycetes; the terrestrial-aquatic hyphomycetes; and the submerged-aquatic hyphomycetes. The adaptations of aquatic fungi for their dispersal and subsequent attachment to new substrates are discussed.     

Elemental composition and degree of homeostasis of fungi: are aquatic hyphomycetes more like metazoans,bacteria or plants?

Ecological stoichiometry generally assumes that heterotrophs have a higher degree of elemental homeostasis than autotrophs. Differences between fixed consumer nutrient requirements and nutrients available in resources allow prediction of the intensity of nutrient recycling ensured by heterotrophs. Despite their fundamental role in detritus decomposition, extremely few data are currently available on fungal elemental composition. In this study, we quantified the degree of elemental homeostasis of aquatic hyphomycetes used as model organisms. Contrary to metazoans, but similar to plants, aquatic hyphomycetes exhibited highly plastic elemental compositions. Mycelium also reached far higher C/nutrient ratios than reported for bacteria. Our results suggest that non-homeostasis of fungi should be explicitly included in stoichiometric models dealing with nutrient recycling, and that the discrepancy in homeostasis between some bacterial strains and fungi should certainly be considered when investigating interactions between both groups of decomposers.     

Comparison of aquatic hyphomycetes communities between lotic and lentic environments in the Atlantic rain forest of Pernambuco,Northeast Brazil

Riparian forests are important to aquatic ecosystems and produce large quantities of organic matter that are recycled by the microbial community that includes microscopic fungi. The aim of this study was to unveil and compare the diversity of aquatic hyphomycetes associated to submerged leaf litter of tropical lotic and lentic environments in the Atlantic Forest of Northeast Brazil. Six sampling events were carried out in six points of two study areas: Biological Reserve “Mata da Chuva” (MC) and Environmental Protection Area “Lagoa da Mata” (LM), in Pernambuco, Brazil. Twenty three taxa of hyphomycetes were identified resulting in 87 occurrences. In the lake LM, 13 taxa of hyphomycetes were identified with 34 occurrences and in the MC (stream), 20 taxa with 53 occurrences. Ten species were common to both areas. Diversity indices and fungal biomass (ergosterol) were mostly higher in the lotic system. The fungal community analysis did not show any structure regarding sampling periods or sampling points within an area, however the two areas are different. Although the turbulence of the water is considered important for the development of these aquatic fungi, it is possible to find a diverse community of hyphomycetes and considerable fungal biomass in the lentic environment.     

Does nanosized plastic affect aquatic fungal litter decomposition?

Nanosized plastics are an emerging concern in freshwater ecosystems, raising the question whether they put freshwater ecological processes at risk. Litter decomposition is a major ecological function in forested streams which is mainly driven by aquatic hyphomycetes. Here we investigated whether increasing concentrations (up to 102.4 mg/L) of nanosized polystyrene plastics (NPPs; 100nm) affect litter decomposition by five widely distributed species of aquatic hyphomycetes. Results showed that average litter decomposition decreased by 8% relative to the control when exposed to 102.4 mg/L NPPs. Aquatic hyphomycete species differed in their sensitivity to NPPs. The greatest inhibition of litter decomposition was found with Tetracladium marchalianum, where it dropped from 37 (control) to 16% (102.4 mg/L of NPP). Overall our study highlights the emerging risks and potential dangers of NPPs to freshwater ecosystem functioning. It also indicates that the impact of NPPs may be species specific.     

Allelopathic inhibition of primary producer growth and photosynthesis by aquatic fungi

Autochthonous primary production is generally much reduced in forested headwater streams. Several hypotheses have been proposed for explaining this observation, among them, the low light intensity, or the strong constraints exerted by stream current. Allelopathic inhibition of competitors is a common ecological process in aquatic environments. Aquatic hyphomycetes are known to chemically inhibit bacteria and other fungi (including other aquatic hyphomycetes) but a possible allelopathic effect of aquatic hyphomycetes on primary producers has never been tested. The inhibitory effect of twelve aquatic hyphomycete species was tested on three diatom species. Nine aquatic hyphomycete species exhibited anti-diatom activity. Up to 100% diatom growth inhibition was observed. Our study reveals that such allelopathic interactions might be common in streams and probably involve an array of fungal compounds. We propose that the generally reduced primary production observed in forested headwater streams is, among other factors, due to the inhibition of primary producers by allelopathic compounds released by aquatic hyphomycetes.     

Effects of Zinc on Leaf Decomposition by Fungi in Streams: Studies in Microcosms

The effect of zinc on leaf decomposition by aquatic fungi was studied in microcosms. Alder leaf disks were precolonized for 15 days at the source of the Este River and exposed to different zinc concentrations during 25 days. Leaf mass loss, fungal biomass (based on ergosterol concentration), fungal production (rates of [1-¹⁴C]acetate incorporation into ergosterol), sporulation rates, and species richness of aquatic hyphomycetes were determined. At the source of the Este River decomposition of alder leaves was fast and 50% of the initial mass was lost in 25 days. A total of 18 aquatic hyphomycete species were recorded during 42 days of leaf immersion. Articulospora tetracladia was the dominant species, followed by Lunulospora curvula and two unidentified species with sigmoid conidia. Cluster analysis suggested that zinc concentration and exposure time affected the structure of aquatic hyphomycete assemblages, even though richness had not been severely affected. Both zinc concentration and exposure time significantly affected leaf mass loss, fungal production and sporulation, but not fungal biomass. Zinc exposure reduced leaf mass loss, inhibited fungal production and affected fungal reproduction by either stimulating or inhibiting sporulation rates. The results of this work suggested zinc pollution might depress leaf decomposition in streams due to changes in the structure and activity of aquatic fungi.     

The molecular phylogeny of aquatic hyphomycetes with affinity to the Leotiomycetes

Aquatic hyphomycetes play a key role in decomposition of submerged organic matter and stream ecosystem functioning. We examined the phylogenetic relationships among various genera of aquatic hyphomycetes belonging to the Leotiomycetes (Ascomycota) using sequences of internal transcribed spacer (ITS) and large subunit (LSU) regions of rDNA generated from 42 pure cultures including 19 ex-types. These new sequence data were analyzed together with additional sequences from 36 aquatic hyphomycetes and 60 related fungi obtained from GenBank. Aquatic hyphomycetes, characterized by their tetraradiate or sigmoid conidia, were scattered in nine supported clades within the Helotiales (Leotiomycetes). Tricladium, Lemonniera, Articulospora, Anguillospora, Varicosporium, Filosporella, and Flagellospora are not monophyletic, with species from the same genus distributed among several major clades. The Gyoerffyella clade and the Hymenoscyphus clade accommodated species from eight and six different genera, respectively. Thirteen aquatic hyphomycete taxa were grouped in the Leotia-Bulgaria clade while twelve species clustered within the Hymenoscyphus clade along with several amphibious ascomycetes. Species of Filosporella and some species from four other aquatic genera were placed in the Ascocoryne-Hydrocina clade. It is evident that many aquatic hyphomycetes have relatives of terrestrial origin. Adaptation to colonize the aquatic environment has evolved independently in multiple phylogenetic lineages within the Leotiomycetes.     

Effects of increased temperature and aquatic fungal diversity on litter decomposition

Climate warming and biodiversity loss are two major factors threatening freshwaters. Aquatic hyphomycetes are fungi that play a key role in organic matter turnover in streams. To assess the impacts of temperature increase and aquatic hyphomycete diversity on plant-litter decomposition, we manipulated fungal assemblage composition at two levels of diversity (four and eight species) under ambient temperature of 16 °C and two regimes of temperature increase differing in 8 °C: abrupt versus gradual increase from 16 to 24 °C. The effects were evaluated on leaf-litter decomposition, fungal biomass and reproduction. Results showed faster leaf decomposition under increased temperature, but no differences were found between an abrupt and a gradual increase in temperature. Assemblage composition was the major factor controlling fungal biomass and reproduction, while fungal diversity was only critical to maintain reproduction.