RESUMO
Laccaria amethystine and L. bicolor have served as model species for studying the life history and genetics of ectomycorrhizal fungi. However, the characterizations and variations of their mitogenomes are still unknown. In the present study, the mitogenomes of the two Laccaria species were assembled, annotated, and compared. The two mitogenomes of L. amethystine and L. bicolor comprised circular DNA molecules, with the sizes of 65,156 bp and 95,304 bp, respectively. Genome collinearity analysis revealed large-scale gene rearrangements between the two Laccaria species. Comparative mitogenome analysis indicated the introns of cox1 genes in Agaricales experienced frequent lost/gain eveants, which promoted the organization and size variations in Agaricales mitogenomes. Evolutionary analysis indicated the core protein-coding genes in the two mitogenomes were subject to strong pressure of purifying selection. Phylogenetic analysis using the Bayesian inference (BI) and Maximum likelihood (ML) methods based on a combined mitochondrial gene set resulted in identical and well-supported tree topologies, wherein the two Laccaria species were most closely related to Coprinopsis cinerea. This study severed as the first study on the mitogenomes of Laccaria species, which promoted a comprehensive understanding of the genetics and evolution of the model ectomycorrhizal fungi.
Assuntos
Genes Mitocondriais/genética , Genoma Mitocondrial/genética , Íntrons/genética , Laccaria/genética , Micorrizas/genética , Filogenia , Agaricales/genética , Basidiomycota/classificação , Basidiomycota/genética , Teorema de Bayes , Complexo IV da Cadeia de Transporte de Elétrons/genética , Proteínas Fúngicas/genética , Rearranjo Gênico , Laccaria/classificação , Micorrizas/classificação , RNA/genética , Análise de SequênciaRESUMO
Species of Laccaria (Hydnangiaceae, Basidiomycota) are important in forest ecosystems as ectomycorrhizal fungi. Nine of the 75 described Laccaria species worldwide been reported from Korea. Most of these have European and North American names, and their identities are based solely on morphological features. To evaluate the taxonomy of Korean Laccaria, we used 443 specimens collected between 1981 and 2016 in a phylogenetic analysis based on sequence data from nuc rDNA internal transcribed spacer ITS1-5.8S-ITS2 rDNA (ITS) region, nuc 28S rDNA (28S), RNA polymerase II subunit 2 (rpb2), and translation elongation factor 1-α (tef1). Ten Laccaria species were identified. Three of these were previously reported from Korea: L. bicolor, L. tortilis, and L. vinaceoavellanea. Laccaria alba, L. japonica, and L. murina are confirmed as new reports from Korea. Lastly, four new Laccaria species are described: L. araneosa, L. parva, L. torosa, and L. versiforma. This study supports the general contention that Asian species of ectomycorrhizal fungi may not be conspecific with morphologically similar species from Europe and North America. Furthermore, identification based on morphology alone is often unreliable in Laccaria due to considerable overlap of characters among species. Thus, use of molecular methods is necessary for effective identification. Illustrations of the four newly described species and a taxonomic key to species of Laccaria in Korea are provided.
Assuntos
Carpóforos/crescimento & desenvolvimento , Laccaria/classificação , Micorrizas/classificação , Filogenia , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Coreia (Geográfico) , Laccaria/citologia , Laccaria/genética , Laccaria/crescimento & desenvolvimento , Microscopia , Microscopia Eletrônica de Varredura , Micorrizas/citologia , Micorrizas/genética , Micorrizas/crescimento & desenvolvimento , Fator 1 de Elongação de Peptídeos/genética , RNA Polimerase II/genética , RNA Ribossômico 28S/genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Esporos Fúngicos/citologia , Esporos Fúngicos/ultraestruturaRESUMO
Purple Laccaria are ectomycorrhizal basidiomycetes associated with temperate forests all over the Northern Hemisphere in at least two taxa: Laccaria amethysteo-occidentalis in North America, and L. amethystina complex in Eurasia, as shown by Vincenot et al. (2012). Here, we combine a further study of the genetic structure of L. amethystina populations from Europe to southwestern China and Japan, using neutral Single Sequence Repeat (SSR; microsatellite) markers; and a systematic description of two novel Asian species, namely Laccaria moshuijun and Laccaria japonica, based on ecological, morphological, and molecular criteria (rDNA sequences). Population genetics provides evidence of the ancient isolation of three regional groups, with strong signal for speciation, and suggests a centre of origin of modern populations closest to present-day Chinese populations. Phylogenetic analyses confirm speciation at the molecular level, reflected in morphological features: L. moshuijun samples (from Yunnan, China) display strongly variable cheilocystidia, while L. japonica samples (from Japan) present distinctive globose to subglobose spores and clavate cheilocystidia. This study of a species complex primarily described with an extremely wide ecological and geographical range sheds new light on the biodiversity and biogeography of ectomycorrhizal fungi.
Assuntos
Laccaria/classificação , Laccaria/isolamento & purificação , Repetições de Microssatélites , Filogenia , Filogeografia , China , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Europa (Continente) , Japão , Laccaria/citologia , Laccaria/genética , Microscopia , Análise de Sequência de DNARESUMO
Some species of Laccaria have been known to contain relatively high levels of arsenic in Europe and are used as edible mushrooms in the southwest China. One population of Laccaria proxima and one population of L. vinaceoavellanea as well as topsoil (0-10 cm) they grew on were collected from natural habitats of Yunnan (SW China), while other samples such as Laccaria mushroom samples without soil were purchased from four different local markets in Yunnan. Concentrations of arsenic were determined in fruit bodies of the mushrooms and in the soils by using atomic fluorescence spectrometry to assess potential health risks of these species. The mean arsenic concentrations in caps were 135, 14.1-143, 5.5 and 130-163 mg kg(-1) dry weight (dw) for Laccaria amethystina, Laccaria laccata, L. proxima and L. vinaceoavellanea, respectively. The mean value for bioconcentration factor of arsenic in caps of L. vinaceoavellanea was 29.1 for soil with arsenic content at 5.6 mg kg(-1) dw, which indicate that L. vinaceoavellanea is an accumulator for arsenic. Caps of L. amethystina, L. laccata and L. vinaceoavellanea consumed at a volume of 300 g fresh weight for a single meal in a week can yield an exposure amount of arsenic at 4.1, 0.42-4.3 and 3.9-4.9 mg, respectively. These values are higher than the limit dose for the intake of inorganic arsenic recommended by the Joint FAO/WHO Expert Committee on Food Additives.
Assuntos
Agaricales/química , Arsênio/análise , Carpóforos/química , Laccaria/química , China , Contaminação de Alimentos/análise , Contaminação de Alimentos/prevenção & controle , Geografia , Laccaria/classificação , Saúde Pública , Medição de Risco , Fatores de Risco , Solo/química , Poluentes do Solo/análise , Especificidade da Espécie , Espectrofotometria AtômicaRESUMO
Given the diversity and ecological importance of Fungi, there is a lack of population genetic research on these organisms. The reason for this can be explained in part by their cryptic nature and difficulty in identifying genets. In addition the difficulty (relative to plants and animals) in developing molecular markers for fungal population genetics contributes to the lack of research in this area. This study examines the ability of restriction-site associated DNA (RAD) sequencing to generate SNPs in Laccaria bicolor. Eighteen samples of morphologically identified L. bicolor from the United States and Europe were selected for this project. The RAD sequencing method produced anywhere from 290 000 to more than 3 000 000 reads. Mapping these reads to the genome of L. bicolor resulted in 84 000-940 000 unique reads from individual samples. Results indicate that incorporation of non-L. bicolor taxa into the analysis resulted in a precipitous drop in shared loci among samples, suggests the potential of these methods to identify cryptic species. F-statistics were easily calculated, although an observable "noise" was detected when using the "All Loci" treatment versus filtering loci to those present in at least 50% of the individuals. The data were analyzed with tests of Hardy-Weinburg equilibrium, population genetic statistics (FIS and FST), and population structure analysis using the program Structure. The results provide encouraging feedback regarding the potential utility of these methods and their data for population genetic analysis. We were unable to draw conclusions of life history of L. bicolor populations from this dataset, given the small sample size. The results of this study indicate the potential of these methods to address population genetics and general life history questions in the Agaricales. Further research is necessary to explore the specific application of these methods in the Agaricales or other fungal groups.
Assuntos
Genoma Fúngico , Laccaria/genética , Laccaria/isolamento & purificação , Análise de Sequência de DNA/métodos , Genótipo , Laccaria/classificação , Dados de Sequência Molecular , FilogeniaRESUMO
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.
Assuntos
Variação Genética , Laccaria/classificação , Laccaria/genética , Micorrizas/classificação , Micorrizas/genética , Esporos Fúngicos/classificação , Esporos Fúngicos/genética , Alelos , Japão , Laccaria/isolamento & purificação , Micorrizas/isolamento & purificação , Densidade Demográfica , Dinâmica Populacional , Esporos Fúngicos/isolamento & purificação , Erupções VulcânicasRESUMO
Laccaria (Hydnangiaceae, Agaricales, Basidiomycota) is one of the more intensively studied ectomycorrhizal genera; however, species boundaries within Laccaria and the closely related Hydnangium and Podohydnangium in Australia have not yet been examined with molecular sequence data. Based on morphological characters, eight native species of Laccaria are currently recognized in Australia, as well as three Hydnangium species and the monotypic Podohydnangium australe. Sequences of the internal transcribed spacer region of nuclear rDNA (ITS), RNA polymerase beta subunit II (rpb2) and translation elongation factor 1 alpha (tef-1α) were generated from 77 collections of Laccaria, Hydnangium and Podohydnangium from Australia. Ten phylogenetic species and a further 11 potential species (represented by singletons) of Laccaria in Australia are delimited from sequence analyses. Most of the morphological species contained cryptic phylogenetic species, but these species were always nested entirely within a given morphological species, although not always as sister taxa. The rpb2 locus performed best as a species barcode with pairwise and patristic distance measures. The ITS sequence region returned the least resolved gene tree of the three regions examined and was the least useful as a barcode region. Based on the phylogenetic topology, there appears to have been multiple gains and/or losses of the ectomycorrhizal association of some species with the myrtle beech, Nothofagus cunninghamii as well as of sequestrate basidiocarps and two-spored basidia.
Assuntos
DNA Fúngico/genética , Laccaria/classificação , Laccaria/genética , DNA Ribossômico/genética , RNA Polimerases Dirigidas por DNA/genética , Dados de Sequência Molecular , Família Multigênica , Elongação Traducional da Cadeia Peptídica/genética , Filogenia , Austrália do SulRESUMO
Hydrophobins are morphogenetic, small secreted hydrophobic fungal proteins produced in response to changing development and environmental conditions. These proteins are important in the interaction between certain fungi and their hosts. In mutualistic ectomycorrhizal fungi several hydrophobins form a subclass of mycorrhizal-induced small secreted proteins that are likely to be critical in the formation of the symbiotic interface with host root cells. In this study, two genomes of the ectomycorrhizal basidiomycete Laccaria bicolor strains S238N-H82 (from North America) and 81306 (from Europe) were surveyed to construct a comprehensive genome-wide inventory of hydrophobins and to explore their characteristics and roles during host colonization. The S238N-H82 L. bicolor hydrophobin gene family is composed of 12 genes while the 81306 strain encodes nine hydrophobins, all corresponding to class I hydrophobins. The three extra hydrophobin genes encoded by the S238N-H82 genome likely arose via gene duplication and are bordered by transposon rich regions. Expression profiles of the hydrophobin genes of L. bicolor varied greatly depending on life stage (e.g. free living mycelium vs. root colonization) and on the host root environment. We conclude from this study that the complex diversity and range of expression profiles of the Laccaria hydrophobin multi-gene family have likely been a selective advantage for this mutualist in colonizing a wide range of host plants.
Assuntos
Proteínas Fúngicas/genética , Laccaria/classificação , Laccaria/genética , Micorrizas/classificação , Micorrizas/genética , Filogenia , Sequência de Aminoácidos , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Regulação Fúngica da Expressão Gênica , Genoma Fúngico , Laccaria/crescimento & desenvolvimento , Laccaria/metabolismo , Dados de Sequência Molecular , Família Multigênica , Micorrizas/crescimento & desenvolvimento , Micorrizas/metabolismo , Alinhamento de SequênciaRESUMO
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.