New findings on the fungal species Tricholoma matsutake from Ukraine, and revision of its taxonomy and biogeography based on multilocus phylogenetic analyses.

Matsutake mushrooms are among the best-known edible wild mushroom taxa worldwide. The representative Tricholoma matsutake is from East Asia and the northern and central regions of Europe. Here, we report the existence of T. matsutake under fir trees in Eastern Europe (i.e., Ukraine), as confirmed by phylogenetic analysis of nine loci on the nuclear and mitochondrial genomes. All specimens from Japan, Bhutan, China, North Korea, South Korea, Sweden, Finland, and Ukraine formed a T. matsutake clade according to the phylogeny of the internal transcribed spacer region. The European population of T. matsutake was clustered based on the ß2 tubulin gene, with a moderate bootstrap value. In contrast, based on analyses of three loci, i.e., rpb2, tef1, and the ß2 tubulin gene, T. matsutake specimens sampled from Bhutan and China belonged to a clade independent of the other specimens of this species, implying a genetically isolated population. As biologically available type specimens of T. matsutake have not been designated since its description as a new species from Japan in 1925, we established an epitype of this fungus, sampled in a Pinus densiflora forest in Nagano, Japan.

Phylogeny of epilachna, henosepilachna, and some minor genera of phytophagous ladybird beetles (coleoptera: coccinellidae: coccinellinae: epilachnini), with an analysis of ancestral biogeography and host-plant utilization.

Ladybird beetles in the tribe Epilachnini include notorious crop pests and model species studied intensively in various fields of evolutionary biology. From a combined dataset of mitochondrial (ND2) and nuclear (28S) DNA sequences, we reconstructed the phylogeny of 46 species of Epilachnini from Asia, Africa, America, and the Australian region: 16 species in Epilachna, 24 species in Henosepilachna, and one species each in Adira, Afidenta, Afidentula, Afissula, Chnootriba, and Epiverta. In our phylogenetic trees, both Epilachna and Henosepilachna were reciprocally polyphyletic. Asian Epilachna species were monophyletic, except for the inclusion of Afissula sp. Asian and Australian Henosepilachna species likewise formed a monophyletic group, excluding H. boisduvali. African Epilachna and Henosepilachna species did not group with their respective Asian and American congeners, but were paraphyletic to other clades (Epilachna species) or formed a separate monophyletic group (Henosepilachna species) together with Chnootriba similis. The American Epilachna species were monophyletic and formed a clade with American Adira clarkii and Asian Afidentula manderstjernae bielawskii; this clade was the sister group to Asian and Australian Henosepilachna, but was distant from Asian Epilachna. Chnootriba was embedded in the African Henosepilachna clade, and Afissula in the Asian Epilachna clade. Epiverta, which is morphologically unique, was the sister group to Asian Epilachna, although with weak support. From reconstructions of biogeographical distribution and host-plant utilization at ancestral nodes, we inferred an African origin for the common ancestor of the species studied, and found the frequency of host shifts to differ greatly between the two major lineages of Epilachnini examined.

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.

Phylogenetic relationships among host plants explain differences in fungal species richness and community composition in ectomycorrhizal symbiosis.

Geographic and taxonomic host ranges determine the distribution of biotrophic organisms. Host phylogenetic distance strongly affects the community composition of pathogens and parasites, but little is known about the host phylogeny effect on communities of mutualists, such as plant-pollinator and plant-mycorrhizal fungi systems. By incorporating phylogenetic eigenvectors into univariate and multivariate models, we aimed to determine the relative contribution of host phylogeny and environmental variables to mycorrhizal traits and community composition of ectomycorrhizal (EcM) fungi in Salicaceae at the local scale. Host phylogeny explained 75% of the variation in fungal species richness and 20% of the variation in community composition. We also re-analyzed a system involving eight hosts from Japan, in which host phylogeny explained 26% and 9% of the variation in fungal richness and community composition, respectively. [Correction added after online publication 21 May 2013: in the preceding sentence the values 9% and 26% have been transposed.] Phylogenetic eigenvectors that differentially account for clades and terminal taxa across the phylogeny revealed stronger host effects than did the treatment of host species as categorical or dummy variables in multiregression models, and in comparison with methods such as Mantel test and its analogs. Our results indicate the usefulness of the eigenvector method for the quantification of the host phylogeny effect, which represents an integrated complex function of taxonomic sampling effect and phylogenetic distance per se.

Ectomycorrhizal fungal community in alkaline-saline soil in northeastern China.

Alkaline-saline soil is widespread in arid and semiarid regions of the world and causes severe environmental and agricultural problems. To advance our understanding of the adaptation of ectomycorrhizal fungi (EMF) to alkaline-saline soil, we investigated EMF communities on Mongolian willow (Salix linearistipularis) growing in alkaline-saline soil (up to pH 9.2) in northeastern China. In total, 75 root samples were collected from 25 willow individuals over 4.7 ha. To identify fungal species in ectomycorrhizal root tips, we used terminal restriction fragment length polymorphism and sequencing analyses of the internal transcribed spacer region of ribosomal DNA. We detected 11 EMF species, including species of Inocybe, Hebeloma, and Tomentella of the Basidiomycota and three Ascomycota species. The EMF richness of the study site was estimated to be 15-17 using major estimators. The most abundant species was Geopora sp. 1, while no Geopora-dominated EMF communities have been reported so far. Phylogenetic analysis showed that the phylogroup including Geopora sp. 1 has been found mostly in alkaline soil habitats, indicating its adaptation to high soil pH. Because EMF are indispensable for host plant growth, the EMF species detected in this study may be useful for restoration of alkaline-saline areas.

Germination and infectivity of ectomycorrhizal fungal spores in relation to their ecological traits during primary succession.

The spores of ectomycorrhizal fungi (EMF) play critical roles in the population and community development of EMF. Here, the germination and infectivity of EMF spores are examined with reference to the ecological traits of the EMF species. Spores were collected from 12 EMF species, whose successional patterns have been studied in the volcanic desert on Mount Fuji, Japan. Spore germination experiments were conducted with host plants (Salix reinii), with nonhost plants (Polygonum cuspidatum), and without plants. The mycorrhizal formation ability of spores was also examined in seven EMF using spore inoculation experiments. To determine the effects of the spore preservation period, both experiments were repeated up to 1 yr after spore collection. Spore germination was very low in the absence of host plants. In the presence of hosts, even 30 d after spore collection, spore germination was significantly enhanced in all pioneer EMF (c. 20%) but less so in late-stage EMF (< 5%), except in Hebeloma species. Mycorrhizal formation from spores was also greater in pioneer EMF but was significantly reduced by 1 yr of spore preservation. High spore germination and infectivity of pioneer EMF should enable these species to colonize disturbed and isolated areas in accordance with their ecological traits.

Host effects on ectomycorrhizal fungal communities: insight from eight host species in mixed conifer-broadleaf forests.

To advance our understanding of host effects on the community structure of ectomycorrhizal fungi (EMF), EMF communities were compared among different host species, genera and families in two mixed conifer-broadleaf forests in Japan. Using molecular identification methods we examined EMF root tips of eight coexisting species belonging to six genera (three families): Abies and Tsuga (Pinaceae), Betula and Carpinus (Betulaceae) and Fagus and Quercus (Fagaceae). In total, 205 EMF species were detected, and the total richness was estimated to exceed 300 species using major estimators. Of the 55 EMF species occurring three or more times, eight showed significantly biased host preference. A Mantel test showed a significantly negative correlation between EMF community similarity and host taxonomic distance. Detrended correspondence analysis separated EMF communities mainly by host taxonomic and successional status. Thus, EMF communities are similar on hosts with similar taxonomic and successional status. A significant proportion of EMF exhibited host specificity, which may contribute to the extremely diverse EMF community in conifer-broadleaf forests.

Sex allocation of a cosexual wind-pollinated tree, Quercus dentata, in terms of four currencies.

Sex allocation of a cosexual wind-pollinated species, Quercus dentata (Fagaceae), was analyzed using biomass, carbon, nitrogen and phosphorus as currencies based on data accumulated for 61 individuals from 1997-2004. Strongly female-biased sex allocation was indicated when measured in terms of biomass and carbon, but no significant bias was detected when measured in terms of nitrogen or phosphorus. From an adaptive viewpoint, there is little support for strong female-biased sex allocation, suggesting that sex allocation in terms of nitrogen or phosphorus is closer to the real picture. The relative sex allocation considerably varied from year to year, but the relative femaleness of individuals in the population was rather constant across years. No significant correlation was observed between relative sex allocation and fecundity or tree height, but individuals that showed very low fecundity tended to produce only acorns.

Differentiation and hybridization between Quercus crispula and Q. dentata (Fagaceae): insights from morphological traits, amplified fragment length polymorphism markers, and leafminer composition.

Quercus crispula and Q. dentata (Fagaceae) are dominant members of cool-temperate forests of Japan and are assumed to hybridize in nature. To characterize and discriminate these two species and their hybrids, we carried out multivariate analysis using several morphological traits and principal coordinate analysis using molecular (amplified fragment length polymorphism [AFLP]) data. Further, we examined the composition of Phyllonorycter species (leafmining insects) on individuals from a mixed forest. Morphological traits and Phyllonorycter composition differ enough in these two oak species to be useful for identification of species and hybrids. AFLP data, however, are less informative because the degree of molecular differentiation between the two species is low. Nine out of 105 individuals from a mixed stand had intermediate morphologies according to the multivariate analysis, and eight out of the nine individuals had intermediate Phyllonorycter composition in either one or both of the two study years. These eight individuals were tentatively assigned as hybrids or backcross individuals, and the remaining individual with intermediate morphologies was assigned as Q. dentata according to its Phyllonorycter composition and the AFLP analysis.