Little evidence for niche partitioning among ectomycorrhizal fungi on spruce seedlings planted in decayed wood versus mineral soil microsites

Ectomycorrhizal fungal (EMF) communities vary among microhabitats, supporting a dominant role for deterministic processes in EMF community assemblage. EMF communities also differ between forest and clearcut environments, responding to this disturbance in a directional manner over time by returning to the species composition of the original forest. Accordingly, we examined EMF community composition on roots of spruce seedlings planted in three different microhabitats in forest and clearcut plots: decayed wood, mineral soil adjacent to downed wood, or control mineral soil, to determine the effect of retained downed wood on EMF communities over the medium and long term. If downed and decayed wood provide refuge habitat distinct from that of mineral soil, we would expect EMF communities on seedlings in woody habitats in clearcuts to be similar to those on seedlings planted in the adjacent forest. As expected, we found EMF species richness to be higher in forests than clearcuts (P ≤ 0.01), even though soil nutrient status did not differ greatly between the two plot types (P ≥ 0.05). Communities on forest seedlings were dominated by Tylospora spp., whereas those in clearcuts were dominated by Amphinema byssoides and Thelephora terrestris. Surprisingly, while substrate conditions varied among microsites (P ≤ 0.03), especially between decayed wood and mineral soil, EMF communities were not distinctly different among microhabitats. Our data suggest that niche partitioning by substrate does not occur among EMF species on very young seedlings in high elevation spruce-fir forests. Further, dispersal limitations shape EMF community assembly in clearcuts in these forests.     

Community analysis of arbuscular mycorrhizal fungi in a warm-temperate deciduous broad-leaved forest and introduction of the fungal community into the seedlings of indigenous woody plants

A community of arbuscular mycorrhizal (AM) fungi was investigated in a warm-temperate deciduous broad-leaved forest using a molecular analysis method. Root samples were obtained from the forest, and DNA was extracted from the samples. Partial 18S rDNA of AM fungi were amplified from the extracted DNA by polymerase chain reaction using a universal eukaryotic primer NS31 and an AM fungal-specific primer AM1. After cloning the PCR products, 394 clones were obtained in total, which were divided into five types by restriction fragment length polymorphism (RFLP) with HinfI, RsaI, and Hsp92II. More than 20% of the clones were randomly selected from each RFLP type and sequenced. Phylogenetic analysis showed that all the obtained clones belonged to Glomus but could not be identified at species level. Topsoil of the forest containing plant roots was inoculated to nonmycorrhizal seedlings of indigenous woody plants, Rhus javanica var. roxburghii and Clethra barvinervis, to introduce the community of AM fungi into the seedlings. Among these five RFLP types, four types were detected from both seedlings, which indicates that the AM fungal community in the forest root samples was introduced at least partly into the seedlings. Meanwhile, an additional four types that were not found in the forest root samples were newly detected in the seedlings, these types were closely related to one another and close to G. fasciculatum or G. intraradices. It is expected that a community of indigenous diverse AM fungi could be introduced into target fields by planting these mycorrhizal seedlings.     

Ectomycorrhizal fungal exoenzyme activity differs on spruce seedlings planted in forests versus clearcuts

Key message

Ectomycorrhizal (ECM) fungal community structure and potential exoenzymatic activity change after clearcut harvesting, but functional complementarity and redundancy among those ECM fungal species remaining support growth of regenerating seedlings.

Abstract

Ectomycorrhizal (ECM) fungal community composition is altered by forest harvesting, but it is not clear if this shift in structure influences ECM fungal physiological function at the community level. In this study, we characterized activities of extracellular enzymes in the ectomycorrhizospheres of Picea engelmannii seedlings grown in forest and clearcut plots. These exoenzymes are critical for the breakdown of large organic molecules, from which nutrients are subsequently absorbed and translocated by ECM fungi to host plants. We found that ectomycorrhizae on seedlings planted in forests had different exoenzyme activity profiles than those on seedlings planted in clearcuts. Specifically, the activities of glucuronidase, laccase, and acid phosphatase were higher on forest seedlings (P ≤ 0.006). These differences may have been partly driven by soil properties. Total carbon, total nitrogen (N), extractable phosphorus, extractable ammonium-N, and mineralizable N were higher, while pH was lower in forest plots (P ≤ 0.01). However, we also found that enzyme activity only shifted where community composition also changed. Functional complementarity can be inferred within ECM fungal communities in both forests and clearcuts because ectomycorrhizae formed by different species in the same environment had distinct enzyme profiles (P < 0.0001). However, ectomycorrhizae of Thelephora terrestris exhibited high levels of N- and P-mobilizing exoenzyme activities. Seedling biomass did not differ between forest and clearcut environments, so the high abundance of T. terrestris ectomycorrhizae in the clearcuts may have sustained nutrient acquisition by clearcut seedlings even in soils with lower N and P and with reduced ECM fungal species richness.

     

Arbuscular mycorrhizal fungal community structures differ between co-occurring tree species of dry Afromontane tropical forest, and their seedlings exhibit potential to trap isolates suited for reforestation

The diversity of arbuscular mycorrhizal (AM) fungi and their broad or narrow association with distinct plant species in natural environments are crucial information in the understanding of the ecological role of AM fungi on plant co-existence. This knowledge is also needed for appropriate mycorrhization of nursery-grown seedlings for forestation efforts. Here, we report results from comparative studies on three co-occurring indigenous tree species of the dry Afromontane forests of Ethiopia and their seedlings grown under controlled conditions in soil collected from the sites. AM fungal SSU rDNA fragment was amplified and sequenced from mycorrhizas of adult plants and seedlings of Olea europaea subsp. cuspidata and Prunus africana, and from Podocarpus falcatus seedlings. AM fungal identity, diversity and community structure were analyzed based on sequence types defined by the NS31-AM1 SSU rDNA fragment similarity in order to compare with data from other habitats. A total of 409 sequences, grouped in 32 sequence types, belonging to Glomeraceae, Diversisporaceae and Gigasporaceae were found. Some sequence types are close to the widespread Glomus intraradices, G. hoi, G. etunicatum, G. cf. etunicatum and Gigaspora margarita. However, the majority (59%) of sequence types are so far specific for the sites including 11 new types when compared with previous data from the same area. The AM fungal community associated with adult plants, including data previously obtained from adult Podocarpus falcatus seedlings, and seedlings of a host species differed significantly, where seedlings trapping a surprising large number of native fungi. AM fungal community structure also differed significantly between host species and sites, respectively. The results confirm previous results from the same area indicating distinct fungal communities associated with the diverse tree species and suggests the potential of these indigenous tree seedlings to trap a wide range of AM fungi appropriate for successful afforestation.     

Molecular diversity of arbuscular mycorrhizal fungi and patterns of host association over time and space in a tropical forest

We have used molecular techniques to investigate the diversity and distribution of the arbuscular mycorrhizal (AM) fungi colonizing tree seedling roots in the tropical forest on Barro Colorado Island (BCI), Republic of Panama. In the first year, we sampled newly emergent seedlings of the understory treelet Faramea occidentalis and the canopy emergent Tetragastris panamensis, from mixed seedling carpets at each of two sites. The following year we sampled surviving seedlings from these cohorts. The roots of 48 plants were analysed using AM fungal-specific primers to amplify and clone partial small subunit (SSU) ribosomal RNA gene sequences. Over 1300 clones were screened for random fragment length polymorphism (RFLP) variation and 7% of these were sequenced. Compared with AM fungal communities sampled from temperate habitats using the same method, the overall diversity was high, with a total of 30 AM fungal types identified. Seventeen of these types have not been recorded previously, with the remainder being similar to types reported from temperate habitats. The tropical mycorrhizal population showed significant spatial heterogeneity and nonrandom associations with the different hosts. Moreover there was a strong shift in the mycorrhizal communities over time. AM fungal types that were dominant in the newly germinated seedlings were almost entirely replaced by previously rare types in the surviving seedlings the following year. The high diversity and huge variation detected across time points, sites and hosts, implies that the AM fungal types are ecologically distinct and thus may have the potential to influence recruitment and host composition in tropical forests.     

Dual colonization of Eucalyptus urophylla S.T. Blake by arbuscular and ectomycorrhizal fungi affects levels of insect herbivore attack

Abstract 1 Eucalypts are an important part of plantation forestry in Asia but, in south China, productivity is very low. This is due to infertile soils and lack of indigenous symbiotic mycorrhizal fungi. The genus Eucalyptus is unusual because it forms both arbuscular (AM) and ectomycorrhizal (ECM) associations. 2 Eucalyptus urophylla saplings were grown with and without AM (Glomus caledonium) and ECM (Laccaria laccata) fungi in a factorial design. Two experiments were performed: one to simulate nursery conditions and the other to simulate the early stages of plantation establishment. Plant growth was measured over 18 weeks and levels of insect attack were recorded. 3 The AM fungus reduced tree growth in the early stages, but the effect appeared to be transient. No effects of ECM were detected on tree growth, but the ectomycorrhiza reduced colonization by the arbuscular mycorrhiza. AM fungi appear to be rapid invaders of the root system, gradually being replaced by ECM. 4 Both fungal types affected levels of damage by insect herbivores. Most importantly, herbivory by the pest insects Anomala cupripes (Coleoptera) and Strepsicrates spp. (Lepidoptera) was decreased by ECM. 5 It is suggested that mycorrhizal effects on eucalypt insects may be determined by carbon allocation within the plant. Future studies of eucalypt mycorrhizas need to take into account the effects of the fungi on foliar‐feeding insects and also the effects of insect herbivory on mycorrhizal establishment.     

Actinomycetes inducing phytotoxic or fungistatic activity in a Douglas-fir Forest and in an adjacent area of repeated regeneration failure in Southwestern Oregon

Actinomycetes were isolated from the upper 1 - 3 cm of the soil layer in a well-developed forest and in an adjacent clearcut area where Douglas-fir [Pseudotsuga menziesii (MIRB.) Franco] regeneration had been impaired for two decades. The population density in the clearcut area was two times as high as that in the forested area. The percentage of actinomycetes that inhibited seed germination of the test plants was significantly higher in isolates obtained from the clearcut area than in those obtained from the forested area, and isolates from the clearcut showed five times the phytotoxic effect of those from the forest. Some actinomycete isolates, 4 % from the clearcut and 2.6 % from the forest, significantly reduced in vitro growth of two common ectomycorrhizal fungi of Douglas-fir,Laccaria laccata andHebeloma ovstuliniforme. Two actinomycete isolates from the clearcut reduced fungal growth by 40 % and 73 %. Reduction of the nutrient in the growth medium did not affect the antifungal activity of the actinomycetes. The data support the idea that, along with other factors, phytotoxic and antifungal actinomycetes may suppress natural regeneration or establishment of planted seedlings - either directly or. indirectly - through inhibition of seed germination or of mycorrhizal fungi.     

Slowed decomposition is biotically mediated in an ectomycorrhizal, tropical rain forest

Bacteria and fungi drive the cycling of plant litter in forests, but little is known about their role in tropical rain forest nutrient cycling, despite the high rates of litter decay observed in these ecosystems. However, litter decay rates are not uniform across tropical rain forests. For example, decomposition can differ dramatically over small spatial scales between low-diversity, monodominant rain forests, and species-rich, mixed forests. Because the climatic patterns and soil parent material are identical in co-occurring mixed and monodominant forests, differences in forest floor accumulation, litter production, and decomposition between these forests may be biotically mediated. To test this hypothesis, we conducted field and laboratory studies in a monodominant rain forest in which the ectomycorrhizal tree Dicymbe corymbosa forms >80% of the canopy, and a diverse, mixed forest dominated by arbuscular mycorrhizal trees. After 2 years, decomposition was significantly slower in the monodominant forest (P < 0.001), but litter production was significantly greater in the mixed forest (P < 0.001). In the laboratory, we found microbial community biomass was greater in the mixed forest (P = 0.02), and the composition of fungal communities was distinct between the two rain forest types (P = 0.001). Sequencing of fungal rDNA revealed a significantly lower richness of saprotrophic fungi in the monodominant forest (19 species) relative to the species-rich forest (84 species); moreover, only 4% percent of fungal sequences occurred in both forests. These results show that nutrient cycling patterns in tropical forests can vary dramatically over small spatial scales, and that changes in microbial community structure likely drive the observed differences in decomposition.     

Response of shrubland birds to regenerating clearcut area and shape

Conservation of birds that breed in early-successional forests will require an understanding of their response to patch size and shape during the early seral stages following disturbance. We modeled the effects of patch area, patch shape, and time period following harvest on territory densities of 5 shrubland bird species and bird community composition in 36 clearcuts from 1997 to 2014 in the Missouri Ozarks, USA. Our best-supported models indicated that densities of indigo bunting (Passerina cyanea), yellow-breasted chat (Icteria virens), and prairie warbler (Setophaga discolor) declined from the early time period (1–7 years post-harvest) to the late time period (12–18 years post-harvest), regardless of clearcut area or shape. Yellow-breasted chat density and bird species richness were positively related to clearcut shape complexity. Bird species richness increased, and bird species density decreased with clearcut area during both time periods. Non-metric multidimensional scaling indicated that bird community composition was strongly related to time since harvest. Within the spatial scale of our study, successional stage is more influential than clearcut size and shape, and some bird species may benefit from relatively small or irregularly shaped clearcuts. © 2019 The Wildlife Society.     

Organic nitrogen use by salal ericoid mycorrhizal fungi from northern Vancouver Island and impacts on growth in vitro of Gaultheria shallon

 Salal (Gaultheria shallon) recovers quickly from rhizomes after clear-cut timber harvesting and dominates clearcuts of Tsuga heterophylla and Thuja plicata forests. Thus it contributes to considerable problems in regeneration of these sites in coastal British Columbia, Canada. Based on what is known about other ericaceous plants, we speculated that mycorrhizal fungi of salal play a vital role in the growth and dominance of salal by providing access to organic nitrogen. In this study, the ability of four species of fungi isolated from salal to use different forms of organic nitrogen was tested in pure culture and in association with salal. The organic forms of nitrogen applied were glutamine (an amino acid), glutathione (a peptide), and bovine serum albumin (BSA, a protein). The fungi tested were Oidiodendron maius, Acremonium strictum, and two nonsporulating fungi. Inoculated plants always grew better than noninoculated plants regardless of nitrogen source. Glutamine was used as readily as ammonium nitrogen by all four fungi and the mycorrhizal plants of salal colonized by those fungi. There was considerable variation between fungus species or the plants inoculated with those fungi in using glutathione and BSA. Salal inoculated with O. maius grew better on glutathione than BSA, while A. strictum and unknown 1 produced significantly greater yields of salal on BSA. Colonization rates of salal by all four fungi was higher on glutathione or BSA than on ammonium or glutamine. Accepted: 19 June 1999     

Patterns of diversity and adaptation in Glomeromycota from three prairie grasslands

Arbuscular mycorrhizal (AM) fungi are widespread root symbionts that often improve the fitness of their plant hosts. We tested whether local adaptation in mycorrhizal symbioses would shape the community structure of these root symbionts in a way that maximizes their symbiotic functioning. We grew a native prairie grass (Andropogon gerardii) with all possible combinations of soils and AM fungal inocula from three different prairies that varied in soil characteristics and disturbance history (two native prairie remnants and one recently restored). We identified the AM fungi colonizing A. gerardii roots using PCR amplification and cloning of the small subunit rRNA gene. We observed 13 operational taxonomic units (OTUs) belonging to six genera in three families. Taxonomic richness was higher in the restored than the native prairies with one member of the Gigaspora dominating the roots of plants grown with inocula from native prairies. Inoculum source and the soil environment influenced the composition of AM fungi that colonized plant roots. Correspondingly, host plants and AM fungi responded significantly to the soil–inoculum combinations such that home fungi often had the highest fitness and provided the greatest benefit to A. gerardii. Similar patterns were observed within the soil–inoculum combinations originating from two native prairies, where five sequence types of a single Gigaspora OTU were virtually the only root colonizers. Our results indicate that indigenous assemblages of AM fungi were adapted to the local soil environment and that this process occurred both at a community scale and at the scale of fungal sequence types within a dominant OTU.     

Nested multiplex PCR—A feasible technique to study partial community of arbuscular mycorrhizal fungi in field-growing plant root

Plant can be infected by different arbuscular mycorrhizal fungi, but little is known about the interaction between them within root tissues mainly because different species cannot be distinguished on the basis of fungal structure. Accurate species identification of Arbuscular mycorrhizal fungi (AMF) colonized in plant roots is the comerstone of mycorrhizal study, yet this fundamental step is impossible through its morphological character alone. For accurate, rapid and inexpensive detection of partial mycorrhizal fungal community in plant roots, a nested multiplex polymerase chain reaction (PCR) was developed in this study. Five discriminating primers designed based on the variable region of the 5′ end of the large ribosomal subunit were used in the experiment for testing their specificity and the sensitivity in nested PCR by using spores from Glomus mosseae (BEG12), Glomus intraradices (BEG141), Scutellospora castaneae (BEG1) and two unidentified Glomus sp. HAUO3 and HAUO4. The feasibility assay of nested multiplex PCR was conducted by use of spore mixture, Astragalus sinicum roots co-inoculated with 4 species of arbuscular mycorrhizal fungi from pot cultures and 15 different field-growing plant roots respectively after analyses of the compatibility of primers. The result indicated that the sensitivity was in the same range as that of the corresponding single PCR reaction. Overall accuracy was 95%. The efficiency and sensitivity of this multiplex PCR procedure provided a rapid and easy way to simultaneously detect several of arbuscular mycorrhizal fungal species in a same plant root system.     

Effects of green tree retention, prescribed burning and soil treatment on pine weevil (Hylobius abietis and Hylobius pinastri) damage to planted Scots pine seedlings

1 Feeding damage and mortality caused to planted Scots pine seedlings by the pine weevils Hylobius abietis and Hylobius pinastri were studied on burned and unburned sites with 0, 10 and 50 m³ per hectare levels of green tree retention from the second to the fourth summer after logging and burning of the sites. 2 The rate of severe feeding damage to pine seedlings caused by pine weevils was higher on burned clearcut sites than on unburned ones, whereas burning did not increase the feeding damage rate on sites with groups of retention trees. The damage rate in the fourth summer was approximately the same on burned and unburned sites. 3 Pine weevil feeding was the major cause of mortality of freshly planted pine seedlings on unburned sites. On burned sites, mortality was higher than the rate of severe feeding damage, particularly in the second summer after burning, possibly owing to fungal attack and abiotic factors. 4 At a retention tree level of 50 m³, feeding damage to the seedlings was lower than on clearcuts and at a 10 m³ retention tree level. Furthermore, on sites with 50 m³ of retention trees, scarification of the soil was found to decrease feeding damage more effectively than on clearcuts and 10 m³ sites. If the seedlings were situated in the centre of scarified patches, scarification alone was as effective as insecticide treatment on unscarified soil for decreasing feeding damage and mortality. 5 The results suggest that when burning is applied as silvicultural treatment after clear‐cuts, retention of trees is recommended to reduce the damages caused by pine weevils on pine seedlings.     

Effect of mycorrhizal inocula on the growth of Sitka spruce seedlings in different soils

Summary Different mycorrhizal fungi were tested for their effectiveness in promoting growth of Sitka spruce seedlings, in two contrasting soils, in a glasshouse pot experiment. In nursery soil,Laccaria amethystina significantly improved growth of seedlings in comparison toL. laccata. Seedlings inoculated with a forest isolate ofThelephora terrestris were significantly larger than those inoculated with a nursery isolate when grown in forest soil. The effectiveness ofComplexipes moniliformis in forest soil was poor in comparison to other mycorrhizal fungi. Strains aswell as species of mycorrhizal fungi affect seedling growth differently. These effects are further influenced by soil type.     

Ectomycorrhizal fungi of Pinus pinea L. in northeastern Spain

 Although Pinus pinea L. is an important forest species in the Mediterranean region, few reports exist on its ectomycorrhizal associates. Sixty isolates, obtained from fungal sporocarps collected in mixed forests of P. pinea in Catalonia (northeastern Spain), were tested for ectomycorrhiza formation on containerized P. pinea seedlings when applied as mycelial inoculum produced in peat-vermiculite. A total of 17 isolates, in 8 genera (Amanita, Hebeloma, Laccaria, Lactarius, Pisolithus, Rhizopogon, Scleroderma and Suillus), formed ectomycorrhizas and the percentages of mycorrhizal short roots varied among isolates and species from 13% to 89%. Some of these fungi are cited for the first time in association with P. pinea. The results indicate further fungal candidates for controlled inoculation of P. pinea seedlings in the nursery. Accepted: 29 October 1998     

Occurrence of arbuscular mycorrhiza in Castanospermum australe A. Cunn. & C. Fraser and effects on growth and production of castanospermine

 Castanospermum australe A. Cunn. & C. Fraser is the only species of the genus Castanospermum (the Moreton Bay chestnut or black bean) native to NE Australia. One constituent of the plant, castanospermine, can inhibit the AIDS virus. The present study investigated possible symbioses between its roots and arbuscular mycorrhizal (AM) fungi. The effects of mycorrhizal fungi on the growth of the plant and yield of alkaloid castanospermine were also studied. The mycorrhizosphere soil and roots of C. australe collected from various sites in and around Sydney, Australia showed AM symbiotic associations with roots, with arbuscules and vesicles in the root cortices. Wet sieving and decanting yielded AM fungal spores, mainly Glomus spp. A positive correlation was found between AM fungal infection and the castanospermine content of seeds of field-grown trees. Field study results were confirmed by growing seedlings under greenhouse conditions and inoculating them with Glomus intraradices Schenck and Smith (INVAM isolate KS906) and Gigaspora margarita Becker & Hall (INVAM isolate BR444–2). The AM fungi increased the growth and P contents of plants and the yield of castanospermine in the leaves, irrespective of the P treatment. No correlation was found between the alkaloid contents of leaves from mycorrhizal seedlings and from non-mycorrhizal plants which received P. No significant difference in the production of castanospermine was found between P treatments when G. margarita was used as inoculum. Accepted: 14 April 1999     

Response of 11 eucalyptus species to inoculation with three arbuscular mycorrhizal fungi

 Numerous publications have reported growth stimulation of Eucalyptus following ectomycorrhizal inoculation in nursery or field conditions. Although Eucalyptus species can also form arbuscular mycorrhiza, their dependency on this type of mycorrhiza is still debatable. This paper presents information on the effect of inoculation of arbuscular mycorrhizal fungi on eucalypt growth. Twenty weeks after mycorrhizal inoculation, Eucalyptus seedlings' stem dry weight could be increased up to 49% compared to non-inoculated control plants. Intensity of root colonization by a given fungus depended on the host species, but it was not related to a plant growth response. Leaf phosphorus concentration of non-inoculated Eucalyptus seedlings varied greatly between species. Increases in leaf phosphorus concentration following mycorrhizal infection were not necessarily associated with plant growth stimulation. The most mycorrhiza-dependent Eucalyptus species tended to be those having the highest leaf phosphorus concentration in the absence of a fungal symbiont. These mycorrhiza-dependent Eucalyptus species seem to have greater phosphorus requirements and consequently to rely more on the symbiotic association. Accepted: 1 September 1995     

Effects of green tree retention, prescribed burning and soil treatment on pine weevil (Hylobius abietis and Hylobius pinastri) damage to planted Scots pine seedlings

1 Feeding damage and mortality caused to planted Scots pine seedlings by the pine weevils Hylobius abietis and Hylobius pinastri were studied on burned and unburned sites with 0, 10 and 50 m³ per hectare levels of green tree retention from the second to the fourth summer after logging and burning of the sites. 2 The rate of severe feeding damage to pine seedlings caused by pine weevils was higher on burned clearcut sites than on unburned ones, whereas burning did not increase the feeding damage rate on sites with groups of retention trees. The damage rate in the fourth summer was approximately the same on burned and unburned sites. 3 Pine weevil feeding was the major cause of mortality of freshly planted pine seedlings on unburned sites. On burned sites, mortality was higher than the rate of severe feeding damage, particularly in the second summer after burning, possibly owing to fungal attack and abiotic factors. 4 At a retention tree level of 50 m³, feeding damage to the seedlings was lower than on clearcuts and at a 10 m³ retention tree level. Furthermore, on sites with 50 m³ of retention trees, scarification of the soil was found to decrease feeding damage more effectively than on clearcuts and 10 m³ sites. If the seedlings were situated in the centre of scarified patches, scarification alone was as effective as insecticide treatment on unscarified soil for decreasing feeding damage and mortality. 5 The results suggest that when burning is applied as silvicultural treatment after clear‐cuts, retention of trees is recommended to reduce the damages caused by pine weevils on pine seedlings.     

Mycetophilidae (Diptera), an insect group vulnerable to forestry practices? A comparison of clearcut,managed and semi-natural spruce forests in southern Norway

The mycetophilid fauna and environmental variables were studied at 15 sites within a spruce forest in southern Norway. There were five replications of each of the following categories: semi-natural forests, clearcuts, and managed forests (clearcut 70–120 years ago). Clearcutting seems to induce a long lasting effect on the Mycetophilidae fauna. The semi-natural forests were more speciesrich and contained more potentially rare species than the two other categories. Even though managed forests and clearcuts differed in faunal composition, their species richness was not significantly different. Continuity is probably a main factor for maintaining the diversity of Mycetophilidae species. Lumping the 15 sites together, the number of species was strongly correlated with the (temporal) continuity of tree cover and substrates, which may reflect an increased diversity of fungal habitats both in dead wood and on the ground. The degree of continuity was recognized by means of indicator species of fungi, lichen and vascular plants. The amount of dead wood was correlated with the species diversity, but was probably dependent on the continuity factor, since clearcuts with much dead wood had relatively fewer species.Important elements in a strategy for conservation of the diversity of mycetophilid species seem to be: (i) to identify and protect the remnant patches of forests with long continuity, (ii) as far as possible, to practice timber harvesting in earlier clearcut forests instead of semi-natural forests.     

Diversity and Spatial Structure of Belowground Plant–Fungal Symbiosis in a Mixed Subtropical Forest of Ectomycorrhizal and Arbuscular Mycorrhizal Plants

Plant–mycorrhizal fungal interactions are ubiquitous in forest ecosystems. While ectomycorrhizal plants and their fungi generally dominate temperate forests, arbuscular mycorrhizal symbiosis is common in the tropics. In subtropical regions, however, ectomycorrhizal and arbuscular mycorrhizal plants co-occur at comparable abundances in single forests, presumably generating complex community structures of root-associated fungi. To reveal root-associated fungal community structure in a mixed forest of ectomycorrhizal and arbuscular mycorrhizal plants, we conducted a massively-parallel pyrosequencing analysis, targeting fungi in the roots of 36 plant species that co-occur in a subtropical forest. In total, 580 fungal operational taxonomic units were detected, of which 132 and 58 were probably ectomycorrhizal and arbuscular mycorrhizal, respectively. As expected, the composition of fungal symbionts differed between fagaceous (ectomycorrhizal) and non-fagaceous (possibly arbuscular mycorrhizal) plants. However, non-fagaceous plants were associated with not only arbuscular mycorrhizal fungi but also several clades of ectomycorrhizal (e.g., Russula) and root-endophytic ascomycete fungi. Many of the ectomycorrhizal and root-endophytic fungi were detected from both fagaceous and non-fagaceous plants in the community. Interestingly, ectomycorrhizal and arbuscular mycorrhizal fungi were concurrently detected from tiny root fragments of non-fagaceous plants. The plant–fungal associations in the forest were spatially structured, and non-fagaceous plant roots hosted ectomycorrhizal fungi more often in the proximity of ectomycorrhizal plant roots. Overall, this study suggests that belowground plant–fungal symbiosis in subtropical forests is complex in that it includes “non-typical” plant–fungal combinations (e.g., ectomycorrhizal fungi on possibly arbuscular mycorrhizal plants) that do not fall within the conventional classification of mycorrhizal symbioses, and in that associations with multiple functional (or phylogenetic) groups of fungi are ubiquitous among plants. Moreover, ectomycorrhizal fungal symbionts of fagaceous plants may “invade” the roots of neighboring non-fagaceous plants, potentially influencing the interactions between non-fagaceous plants and their arbuscular-mycorrhizal fungal symbionts at a fine spatial scale.