Habitat fragmentation strongly restricts gene flow in endangered ectomycorrhizal fungal populations: Evidence from Rhizopogon togasawarius, specific to Pseudotsuga japonica, across the entire distribution range.

Habitat fragmentation reduces gene flow, causing genetic differentiation and diversity loss in endangered species through genetic drift and inbreeding. However, the impact of habitat fragmentation on ectomycorrhizal (ECM) fungi remains unexplored, despite their critical roles in forest ecosystems. Here, we investigated the population genetic structure and the demographic history of Rhizopogon togasawarius, the ECM fungus specifically colonizing the host tree Pseudotsuga japonica, across its entire distribution range (>200 km). These two species are designated as endangered species on the IUCN Red List since they are found only in small, fragmented forests in Japan. We analysed 236 R. togasawarius individuals from five remaining populations across the Kii Peninsula and the Shikoku Island, separated by a sea channel. Simple sequence repeat analyses using 20 loci revealed strong genetic differentiation among populations (FST = 0.255), even significant in the nearest population pair separated by a distance of only 8 km (FST = 0.075), indicating extremely limited gene flow between populations. DIYABC-RF analyses implied that population divergence occurred approximately 6000 generations ago between the two regions, and nearly 1500 generations ago between the nearest populations within Shikoku Island, related to past climate events. Because of prolonged genetic isolation, significant inbreeding was confirmed in four of five populations, where effective population sizes became very small (Ne = 9.0-58.0). Although evaluation of extinction risks for microorganisms is challenging, our conservation genetic results indicated that habitat fragmentation increases extinction risk through population genetic mechanisms, and therefore should not be overlooked in biodiversity conservation efforts.

Effects of root-colonizing fungi on pioneer Pinus thunbergii seedlings in primary successional volcanic mudflow on Kuchinoerabu Island, Japan.

Root-colonizing fungi, such as mycorrhizal fungi and dark septate endophyte fungi, are often found on pioneer plant species during early primary succession. However, little is known about which fungal species are responsible for the establishment of pioneer plants when these symbionts colonize simultaneously. We investigated the root-colonizing fungal communities of Pinus thunbergii that established prior to lichens, bryophytes, and short-lived herbaceous plants in a primary successional volcanic mudflow site on Kuchinoerabu Island, Japan. We collected a total of 54 current-year and 1- to 2-year-old seedlings. The colonization of root fungi was evaluated by direct observation of key structures (e.g., mantle, arbuscule, microsclerotia, and hyphae) and molecular analysis. Of the 34 current-year seedlings collected, only 12 individuals were colonized by ectomycorrhizal (ECM) fungi. By contrast, all 1- to 2-year-old seedlings were colonized by ECM fungi. Seedlings colonized by pine-specific ECM fungi, specifically Rhizopogon roseolus and Suillus granulatus, showed higher nitrogen and phosphorus contents in their needles compared to non-ECM seedlings. Arbuscular mycorrhizal fungi and dark septate endophyte fungi were found in only two and three individuals, respectively. The high density of mycophagous deer on Kuchinoerabu-jima may contribute to the favored dispersal of ECM fungi over other root-colonizing fungi. In conclusion, the seedling establishment of P. thunbergii at the volcanic mudflow may be largely supported by ECM fungi, with negligible effects of arbuscular mycorrhizal fungi and dark septate endophytes.

Primary succession of ectomycorrhizal fungi associated with Alnus sieboldiana on Izu-Oshima Island, Japan.

The primary succession of ectomycorrhizal (ECM) fungi has been well described for Pinus and Salix, but the succession for other pioneer hosts is almost unknown. Here, we investigated ECM fungal communities of Alnus sieboldiana at different host growth stages in a primary successional volcanic site on Izu-Oshima Island, Japan. ECM root tips were collected from 120 host individuals, encompassing seedling, sapling, and mature tree stages. The taxonomic identity of the ECM fungi was determined based on rDNA internal transcribed spacer region sequences. Nine molecular taxonomic units were detected from a total of 807 root tips. The initial ECM fungal community on the pioneer seedlings was composed of only three species, where an undescribed Alpova species (Alpova sp.) was exclusively frequent. With host growth, other ECM fungal species were added to the communities, while the initial colonizers remained even at mature tree stages. Thus, the ECM fungal composition significantly changed along host growth stages and showed the nested community structure. Although most of the ECM fungi confirmed in this study had a broad Holarctic geographical distribution, the Alpova sp. had no previous records in other regions. These results suggest that a locally evolved Alpova sp. plays an essential role in the initial seedling establishment of A. sieboldiana at early successional volcanic sites.

Towards the conservation of ectomycorrhizal fungi on endangered trees: native fungal species on Pinus amamiana are rarely conserved in trees planted ex situ.

Ectomycorrhizal (ECM) symbiosis is essential for the survival of both host trees and associated ECM fungi. However, during conservation activities of endangered tree species, their ECM symbionts are largely ignored. Here, we investigated ECM fungi in ex situ populations established for the conservation of Pinus amamiana, an endangered species distributed on Yakushima Island, Japan. Our objective was to determine whether ECM fungi in natural forests are conserved in ex situ populations on the same island. In particular, we focused on the existence of Rhizopogon yakushimensis, which is specific to P. amamiana and the most dominant in natural P. amamiana forests. Molecular identification of ECM fungi in resident tree roots and soil propagule banks indicated that ECM fungal species native to natural forests were rarely conserved in ex situ populations. Furthermore, R. yakushimensis was not confirmed in any of the resident root or spore bioassay samples from the ex situ populations. Thus, ECM fungal spores may not be effectively dispersed from natural forests located on the same island. Instead, ECM fungi distributed in other geographical regions occurred more frequently in the ex situ populations, indicating unintentional introductions of non-native ECM fungi from the nurseries where seedlings were raised before transplanting. These findings imply that the current ex situ conservation practices of endangered tree do not work for the conservation of native ECM fungi, and instead may need modification to avoid the risk of introducing non-native ECM fungi near the endangered forest sites.

Temperature niche position and breadth of ectomycorrhizal fungi: Reduced diversity under warming predicted by a nested community structure.

Species with narrow niche breadths are assumed to be more susceptible to environmental changes than those with wide niche breadths. Although information on niche properties is necessary for predicting biological responses to environmental changes, such information is largely missing for soil microbes. In this study, we present the temperature niche positions and breadths of a functionally important group of eukaryotic soil microbes, ectomycorrhizal (EM) fungi. We compiled high-quality EM fungal sequence data from 26 forested sites in Japan (with mean annual temperatures ranging from 1.6 to 23.6°C) to create temperature niche profiles for each individual fungal species. Nested theory and a newly developed weighted-randomization null model were applied to 75 fungal operational taxonomic units (OTUs) with high occurrence records to examine potential preferences for certain temperature positions and breadths. Our analyses revealed that (a) many EM fungal OTUs were restricted to habitats with low mean annual temperatures, (b) fungal OTUs observed at colder sites exhibited narrower temperature breadths than expected by chance, (c) the composition of EM fungal OTUs exhibited a nested pattern along the temperature gradient, and (d) EM fungal richness was highest at colder sites, where the greatest degree of overlap in OTU occurrence was observed. These findings imply that future warming may limit the distribution of many EM fungal species that are currently adapted to only cold climates. This could eventually reduce EM fungal biodiversity, which is linked to forest function through symbiotic associations with trees. This study demonstrates the distribution and environmental ranges of various EM fungal species and can contribute to develop species distribution models with the aim of conserving microbes in the face of climate change.

Ectomycorrhizal fungal communities in alpine relict forests of Pinus pumila on Mt. Norikura, Japan.

Ectomycorrhizal (ECM) symbioses are indispensable for the establishment of host trees, yet available information of ECM symbiosis in alpine forests is scarce. Pinus pumila is a typical ice age relict tree species in Japan and often forms monodominant dwarf vegetation above the tree line in mountains. We studied ECM fungi colonizing P. pumila on Mt. Norikura, Japan, with reference to host developmental stages, i.e., from current-year seedlings to mature trees. ECM fungal species were identified based on rDNA ITS sequences. Ninety-two ECM fungal species were confirmed from a total of 2480 root tips examined. Species in /suillus-rhizopogon and /wilcoxina were dominant in seedling roots. ECM fungal diversity increased with host development, due to the addition of species-rich fungal lineages (/cenococcum, /cortinarius, and /russula-lactarius) in late-successional stages. Such successional pattern of ECM fungi is similar to those in temperate pine systems, suggesting the predominant role of /suillus-rhizopogon in seedling establishment, even in relict alpine habitats fragmented and isolated for a geological time period. Most of the ECM fungi detected were also recorded in Europe or North America, indicating their potential Holarctic distribution and the possibility of their comigration with P. pumila through land bridges during ice ages. In addition, we found significant effects of soil properties on ECM fungal communities, which explained 34.1% of the total variation of the fungal communities. While alpine vegetation is regarded as vulnerable to the ongoing global warming, ECM fungal communities associated with P. pumila could be altered by the edaphic change induced by the warming.

Soil spore bank communities of ectomycorrhizal fungi in endangered Chinese Douglas-fir forests.

Chinese Douglas-fir (Pseudotsuga sinensis) is an endangered Pinaceae species found in several isolated regions of China. Although soil spore banks of ectomycorrhizal (ECM) fungi can play an important role in seedling establishment after disturbance, such as in the well-known North American relative (Pseudotsuga menziesii), we have no information about soil spore bank communities in relict forests of Chinese Douglas-fir. We conducted bioassays of 73 soil samples collected from three Chinese Douglas-fir forests, using North American Douglas-fir as bait seedlings, and identified 19 species of ECM fungi. The observed spore bank communities were significantly different from those found in ECM fungi on the roots of resident trees at the same sites (p = 0.02). The levels of potassium (K), nitrogen (N), organic matter, and the pH of soil were the dominant factors shaping spore bank community structure. A new Rhizopogon species was the most dominant species in the spore banks. Specifically, at a site on Sanqing Mountain, 22 of the 57 surviving bioassay seedlings (representing 21 of the 23 soil samples) were colonized by this species. ECM fungal richness significantly affected the growth of bioassay seedlings (R 2 = 0.20, p = 0.007). Growth was significantly improved in seedlings colonized by Rhizopogon or Meliniomyces species compared with uncolonized seedlings. Considering its specificity to Chinese Douglas-fir, predominance in the soil spore banks, and positive effect on host growth, this new Rhizopogon species could play critical roles in seedling establishment and forest regeneration of endangered Chinese Douglas-fir.

Dictyobacter aurantiacus gen. nov., sp. nov., a member of the family Ktedonobacteraceae, isolated from soil, and emended description of the genus Thermosporothrix.

A mesophilic, Gram-stain-positive, spore-forming bacterium that formed branched mycelia was isolated from paddy soil in Gunung Salak (Mount Salak), West Java, Indonesia. This strain, designated S-27T, grew at temperatures between 20 and 37 °C; the optimum growth temperature was 25 to 30 °C, and no growth was observed at 15 or 45 °C. The pH range for growth was pH 3.5 to 8.6; the optimum pH was 6.0, and no growth was observed at pH 3.0 or 9.2. Strain S-27T was able to hydrolyse polysaccharides such as starch, cellulose and xylan. The G+C content of the DNA of strain S-27T was 55.7 mol%. The major fatty acids were iso-C17 : 0 and C16 : 1 2-OH, and the major menaquinone was MK-9 (H2). The cell wall of strain S-27T contained d-glutamic acid, glycine, l-alanine, d-alanine, l-ornithine and ß-alanine in a molar ratio of 1.0 : 1.6 : 1.4 : 0.6 : 0.9 : 1.1. The polar lipids consisted of phosphatidylglycerol, phosphatidylinositol and two glycolipids. The major cell-wall sugar was arabinose. Detailed phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S-27T belongs to the order Ktedonobacterales and is most closely related to Ktedonobacter racemifer SOSP1-21T (89.6 % sequence identity). On the basis of its chemotaxonomic and phenotypic features and phylogenetic position, we concluded that strain S-27T represents a novel genus and species, for which we propose the name Dictyobacter aurantiacus gen. nov., sp. nov. The type strain of Dictyobacter aurantiacus is strain S-27T (=NBRC 109595T=InaCC B312T). Emendation of the description of the genus Thermosporothrix is also provided.

Root-associated fungal communities in three Pyroleae species and their mycobiont sharing with surrounding trees in subalpine coniferous forests on Mount Fuji, Japan.

Pyroleae species are perennial understory shrubs, many of which are partial mycoheterotrophs. Most fungi colonizing Pyroleae roots are ectomycorrhizal (ECM) and share common mycobionts with their Pyroleae hosts. However, such mycobiont sharing has neither been examined in depth before nor has the interspecific variation in sharing among Pyroleae species. Here, we examined root-associated fungal communities in three co-existing Pyroleae species, including Pyrola alpina, Pyrola incarnata, and Orthilia secunda, with reference to co-existing ECM fungi on the surrounding trees in the same soil blocks in subalpine coniferous forests. We identified 42, 75, and 18 fungal molecular operational taxonomic units in P. alpina, P. incarnata, and O. secunda roots, respectively. Mycobiont sharing with surrounding trees, which was defined as the occurrence of the same mycobiont between Pyroleae and surrounding trees in each soil block, was most frequent among P. incarnata (31 of 44 plants). In P. alpina, sharing was confirmed in 12 of 37 plants, and the fungal community was similar to that of P. incarnata. Mycobiont sharing was least common in O. secunda, found in only 5 of 32 plants. Root-associated fungi of O. secunda were dominated by Wilcoxina species, which were absent from the surrounding ECM roots in the same soil blocks. These results indicate that mycobiont sharing with surrounding trees does not equally occur among Pyroleae plants, some of which may develop independent mycorrhizal associations with ECM fungi, as suggested in O. secunda at our research sites.

Ectomycorrhizal fungi reduce the light compensation point and promote carbon fixation of Pinus thunbergii seedlings to adapt to shade environments.

We examined the effects of three ectomycorrhizal (ECM) symbionts on the growth and photosynthesis capacity of Japanese black pine (Pinus thunbergii) seedlings and estimated physiological and photosynthetic parameters such as the light compensation point (LCP), biomass, and phosphorus (Pi) concentration of P. thunbergii seedlings. Through this investigation, we documented a new role of ectomycorrhizal (ECM) fungi: enhancement of the survival and competitiveness of P. thunbergii seedlings under low-light condition by reducing the LCP of seedlings. At a CO2 concentration of 400 ppm, the LCP of seedlings with ECM inoculations was 40-70 µmol photons m-2 s-1, significantly lower than that of non-mycorrhizal (NM) seedlings (200 µmol photons m-2 s-1). In addition, photosynthetic carbon fixation (Pn) increased with light intensity and CO2 level, and the Pn of ECM seedlings was significantly higher than that of NM seedlings; Pisolithus sp. (Pt)- and Laccaria amethystea (La)-mycorrhizal seedlings had significantly lower Pn than Cenococcum geophilum (Cg)-mycorrhizal seedlings. However, La-mycorrhizal seedlings exhibited the highest fresh weight, relative water content (RWC), and the lowest LCP in the mycorrhizal group. Concomitantly, ECM seedlings showed significantly increased chlorophyll content of needles and higher Pi concentrations compared to NM seedlings. Overall, ECM symbionts promoted growth and photosynthesis while reducing the LCP of P. thunbergii seedlings. These findings indicate that ECM fungi can enhance the survival and competitiveness of host seedlings under low light.

Soil propagule banks of ectomycorrhizal fungi share many common species along an elevation gradient.

We conducted bioassay experiments to investigate the soil propagule banks of ectomycorrhizal (EM) fungi in old-growth forests along an elevation gradient and compared the elevation pattern with the composition of EM fungi on existing roots in the field. In total, 150 soil cores were collected from three forests on Mt. Ishizuchi, western Japan, and subjected to bioassays using Pinus densiflora and Betula maximowicziana. Using molecular analyses, we recorded 23 EM fungal species in the assayed propagule banks. Eight species (34.8 %) were shared across the three sites, which ranged from a warm-temperate evergreen mixed forest to a subalpine conifer forest. The elevation pattern of the assayed propagule banks differed dramatically from that of EM fungi on existing roots along the same gradient, where only a small proportion of EM fungal species (3.5 %) were shared across sites. The EM fungal species found in the assayed propagule banks included many pioneer fungal species and composition differed significantly from that on existing roots. Furthermore, only 4 of 23 species were shared between the two host species, indicating a strong effect of bioassay host identity in determining the propagule banks of EM fungi. These results imply that the assayed propagule bank is less affected by climate compared to EM fungal communities on existing roots. The dominance of disturbance-dependent fungal species in the assayed propagule banks may result in higher ecosystem resilience to disturbance even in old-growth temperate forests.

Soil propagule banks of ectomycorrhizal fungi along forest development stages after mining.

Ectomycorrhizal fungal (EMF) propagules play an important role in seedling establishment following disturbance. However, little is known about how the EMF propagule community changes with forest development. In this study, EMF propagules were examined using seedling bioassays in rhizosphere soils collected from a recently closed Pb-Zn tailing (Taolin Pb-Zn tailing (TLT)), a Cu tailing (Dexing Cu No. 2 tailing (DXT)) that had undergone 21 years of restoration, and a mature Masson pine (Pinus massoniana) forest (DXC) outside the Cu mining areas. The corresponding EMF communities colonizing Masson pine at each site were also investigated for comparison. After 8 months of running bioassays, ectomycorrhizal colonization was poor for seedlings grown in TLT (9.0 % ± 14.9 %) and DXT soils (22.4 % ± 17.7 %), while DXC seedlings were well colonized (47.5 % ± 24.9 %). Internal transcribed spacer sequencing revealed that EMF species richness increased with forest development in both the propagule bank (TLT, 6; DXT, 7; DXC, 12) and in the field (TLT, 8; DXT, 14; DXC, 26), though richness was lower in propagule banks. Several lineages, such as Cenococcum, Rhizopogon, Inocybe, Suillus, and Atheliaceae, were frequently encountered in propagule communities, but species assemblages were different among the three sites. Canonical correspondence analysis revealed that several soil parameters, i.e., N, EC, Cu, Pb, Zn, etc., were responsible for the distribution of EMF in the field and bioassay seedlings. The highest overlap in EMF species composition between the propagule bank and the field community was observed at the recently closed tailing (Morisita-Horn similarity = 0.71 for TLT), whereas the lowest overlap occurred at the mature forest (0.26 for DXC). These results indicate that EMF propagules in soil are less frequent and diverse in early primary succession and become more frequent and diverse along forest development, due mainly to the accumulation of dormant spores of Rhizopogon spp. and sclerotia of Cenococcum spp. Thus, EMF propagule communities in soil may diverge from those root-colonizing EMF communities along a gradient of forest development.

Growth and photosynthetic responses of ectomycorrhizal pine seedlings exposed to elevated Cu in soils.

It is still controversial whether ectomycorrhizal (ECM) mycelia filter out toxic metals in nutrient absorption of host trees. In this study, pine (Pinus densiflora) seedlings colonized by Cu-sensitive and Cu-tolerant ECM species were exposed to a wide spectrum of soil Cu concentrations to investigate functions of ECM fungi under Cu stress. The photosynthetic rates of intact needles were monitored in situ periodically. The biomass and elements of plants were also measured after harvest. The ameliorating effect of ECM infection on host plants exposed to toxic stress was metal concentration specific. Under lower-level Cu stress, ECM fungi increased seedling performance, while ECM seedlings accumulated more Cu than nonmycorrhizal (NM) seedlings. Under higher-level Cu stress, photosynthesis decreased well before visible symptoms of Cu toxicity appeared. The reduced photosynthesis and biomass in ECM seedlings compared to NM seedlings under higher Cu conditions were also accompanied by lower phosphorus in needles. There was no marked difference between the two fungal species. Our results indicate that the two ECM fungi studied in our system may not have an ability to selectively eliminate Cu in nutrient absorption and may not act as effective barriers that decrease toxic metal uptake into host plants.

Genet dynamics and ecological functions of the pioneer ectomycorrhizal fungi Laccaria amethystina and Laccaria laccata in a volcanic desert on Mount Fuji.

To understand the reproduction of the pioneer ectomycorrhizal fungi Laccaria amethystina and Laccaria laccata in a volcanic desert on Mount Fuji, Japan, the in situ genet dynamics of sporocarps were analysed. Sporocarps of the two Laccaria species were sampled at fine and large scales for 3 and 2 consecutive years, respectively, and were genotyped using microsatellite markers. In the fine-scale analysis, we found many small genets, the majority of which appeared and disappeared annually. The high densities and annual renewal of Laccaria genets indicate frequent turnover by sexual reproduction via spores. In the large-scale analysis, we found positive spatial autocorrelations in the shortest distance class. An allele-clustering analysis also showed that several alleles were distributed in only a small, localised region. These results indicate that Laccaria spores contributing to sexual reproduction may be dispersed only short distances from sporocarps that would have themselves been established via rare, long-distance spore dispersal. This combination of rare, long-distance and frequent, short-distance Laccaria spore dispersal is reflected in the establishment pattern of seeds of their host, Salix reinii.

Biogeography of ectomycorrhizal fungi associated with alders (Alnus spp.) in relation to biotic and abiotic variables at the global scale.

· Much of the macroecological information about microorganisms is confounded by the lack of standardized methodology, paucity of metadata and sampling effect of a particular substrate or interacting host taxa. · This study aims to disentangle the relative effects of biological, geographical and edaphic variables on the distribution of Alnus-associated ectomycorrhizal (ECM) fungi at the global scale by using comparable sampling and analysis methods. · Ribosomal DNA sequence analysis revealed 146 taxa of ECM fungi from 22 Alnus species across 96 sites worldwide. Use of spatial and phylogenetic eigenvectors along with environmental variables in model selection indicated that phylogenetic relations among host plants and geographical links explained 43 and 10%, respectively,in ECM fungal community composition, whereas soil calcium concentration positively influenced taxonomic richness. · Intrageneric phylogenetic relations among host plants and regional processes largely account for the global biogeographic distribution of Alnus-associated ECM fungi. The biogeography of ECM fungi is consistent with ancient host migration patterns from Eurasia to North America and from southern Europe to northern Europe after the last glacial maximum, indicating codispersal of hosts and their mycobionts.

Revisiting the host effect on ectomycorrhizal fungal communities: implications from host-fungal associations in relict Pseudotsuga japonica forests.

Host identity is among the most important factors in structuring ectomycorrhizal (ECM) fungal communities. Both host-fungal coevolution and host shifts can account for the observed host effect, but their relative significance in ECM fungal communities is not well understood. To investigate these two host-related mechanisms, we used relict forests of Pseudotsuga japonica, which is an endangered endemic species in Japan. As with other Asian Pseudotsuga species, P. japonica has been isolated from North American Pseudotsuga spp. since the Oligocene and has evolved independently as a warm-temperate species. We collected 100 soil samples from four major localities in which P. japonica was mixed with other conifers and broadleaf trees. ECM tips in the soil samples were subjected to molecular analyses to identify both ECM fungi and host species. While 136 ECM fungal species were identified in total, their communities were significantly different between host groups, confirming the existence of the host effect on ECM fungal communities. None of the 68 ECM fungal species found on P. japonica belonged to Pseudotsuga-specific lineages (e.g., Rhizopogon and Suillus subgroups) that are common in North America. Most of ECM fungi on P. japonica were shared with other host fungi or phylogenetically close to known ECM fungi on other hosts in Asia. These results suggest that after migrating, Pseudotsuga-specific fungal lineages may have become extinct in small isolated populations in Japan. Instead, most of the ECM fungal symbionts on P. japonica likely originated from host shifts in the region.

Extensive gene flow over Europe and possible speciation over Eurasia in the ectomycorrhizal basidiomycete Laccaria amethystina complex.

Biogeographical patterns and large-scale genetic structure have been little studied in ectomycorrhizal (EM) fungi, despite the ecological and economic importance of EM symbioses. We coupled population genetics and phylogenetic approaches to understand spatial structure in fungal populations on a continental scale. Using nine microsatellite markers, we characterized gene flow among 16 populations of the widespread EM basidiomycete Laccaria amethystina over Europe (i.e. over 2900 km). We also widened our scope to two additional populations from Japan (10(4) km away) and compared them with European populations through microsatellite markers and multilocus phylogenies, using three nuclear genes (NAR, G6PD and ribosomal DNA) and two mitochondrial ribosomal genes. European L. amethystina populations displayed limited differentiation (average F(ST) = 0.041) and very weak isolation by distance (IBD). This panmictic European pattern may result from effective aerial dispersal of spores, high genetic diversity in populations and mutualistic interactions with multiple hosts that all facilitate migration. The multilocus phylogeny based on nuclear genes confirmed that Japanese and European specimens were closely related but clustered on a geographical basis. By using microsatellite markers, we found that Japanese populations were strongly differentiated from the European populations (F(ST) = 0.416), more than expected by extrapolating the European pattern of IBD. Population structure analyses clearly separated the populations into two clusters, i.e. European and Japanese clusters. We discuss the possibility of IBD in a continuous population (considering some evidence for a ring species over the Northern Hemisphere) vs. an allopatric speciation over Eurasia, making L. amethystina a promising model of intercontinental species for future studies.

Towards global patterns in the diversity and community structure of ectomycorrhizal fungi.

Global species richness patterns of soil micro-organisms remain poorly understood compared to macro-organisms. We use a global analysis to disentangle the global determinants of diversity and community composition for ectomycorrhizal (EcM) fungi-microbial symbionts that play key roles in plant nutrition in most temperate and many tropical forest ecosystems. Host plant family has the strongest effect on the phylogenetic community composition of fungi, whereas temperature and precipitation mostly affect EcM fungal richness that peaks in the temperate and boreal forest biomes, contrasting with latitudinal patterns of macro-organisms. Tropical ecosystems experience rapid turnover of organic material and have weak soil stratification, suggesting that poor habitat conditions may contribute to the relatively low richness of EcM fungi, and perhaps other soil biota, in most tropical ecosystems. For EcM fungi, greater evolutionary age and larger total area of EcM host vegetation may also contribute to the higher diversity in temperate ecosystems. Our results provide useful biogeographic and ecological hypotheses for explaining the distribution of fungi that remain to be tested by involving next-generation sequencing techniques and relevant soil metadata.

Ectomycorrhizal fungal communities associated with Masson pine (Pinus massoniana Lamb.) in Pb-Zn mine sites of central south China.

To advance our understanding of ectomycorrhizal fungal communities in mining areas, the diversity and composition of ectomycorrhizal fungi associated with Masson pine (Pinus massoniana Lamb.) and soil chemistry were investigated in Taolin lead-zinc (Pb-Zn) mine tailings (TLT), two fragmented forest patches in a Huayuan Pb-Zn mineland (HY1 and HY2), and a non-polluted forest in Taolin in central south China. Ectomycorrhizal fungal species were identified by morphotyping and sequence analyses of the internally transcribed spacer regions of ribosomal DNA. The two study sites in the Huayuan mineland (HY1 and HY2) were significantly different in soil Pb, Zn, and cadmium (Cd) concentrations, but no significant difference was observed in ectomycorrhizal colonization, ectomycorrhizal fungal richness, diversity, or rank-abundance. In addition, the similarity of ectomycorrhizal fungal communities between HY1 and HY2 was quite high (Sørensen similarity index = 0.47). Thus, the concentration of heavy metals may not be determining factors in the structure of these communities. In the tailings, however, significantly lower ectomycorrhizal colonization and ectomycorrhizal fungal richness were observed. The amounts of Pb and Zn in the tailing sand were higher than the non-polluted forest but far lower than in HY1. Thus, these heavy metals did not account for the reduced colonization and ectomycorrhizal fungal richness in TLT. The ectomycorrhizal fungal community in TLT was dominated by four pioneer species (Rhizopogon buenoi, Tomentella ellisii, Inocybe curvipes, and Suillus granulatus), which collectively accounted for 93.2 % of root tip colonization. The immature soil conditions in tailing (low N and P, sand texture, and lack of organic matter) may only allow certain pioneer ectomycorrhizal fungal species to colonize the site. When soil samples from four sites were combined, we found that the occurrences of major ectomycorrhizal fungal taxa were not clearly related to the concentrations of Pb, Zn, and Cd. In conclusion, our results suggest that ectomycorrhizal fungal communities in mining areas are not necessarily affected by heavy metals themselves but could be largely determined by soil maturity.