The genus Akanthomyces (Ascomycota, Hypocreales) includes entomopathogenic species known to infect a variety of insects and spiders. In this study, we present the first isolate of A. ampullifer characterized by molecular methods, found on dead bodies of the common cave limoniid Limonia nubeculosa (Diptera) in the subterranean spaces of southwestern Germany. In total, seven specimens exhibited distinctive morphological traits that, when compared with historical records, confirm their identification as A. ampullifer―particularly noted for its affinity to dipteran hosts. Absent from culture collections and molecular repositories, this species has eluded detailed scientific documentation using modern methods. Our research bridges this knowledge gap, providing the first genetic identification barcodes of five genes, living culture, cultivation requirements, and an updated description. This overlooked fungus is phylogenetically most closely related to the species A. pyralidarum, A. laosensis, and some other species mostly associated with adult moths. It demonstrates a unique morphological signature with monoblastic phialides forming a layer on the surface of synnemata and produces long, cylindrical, chain-forming conidia. It prefers lower temperatures, exhibiting an inability to grow at 25 °C, coupled with notably slow growth in culture.
We collected in Japan five sporocarpic specimens morphologically identical to those of Epigeocarpum japonicum, a recently described Glomeromycota species. Although 18S-ITS-28S nuc rDNA sequences obtained from these sporocarps showed high sequence variability, phylogenetic analyses based on 18S-ITS-28S, the largest subunit of the RNA polymerase II (rpb1) gene, and concatenated sequences of the two loci convincingly demonstrated the identity of these sporocarps to E. japonicum. Importantly, the 18S-ITS-28S+rpb1 analyses highlighted the key role of rpb1 sequences in reconstructing the phylogenies of Glomeromycota taxa with strongly divergent rDNA sequences. Upon inoculation with sporocarpic spores, E. japonicum formed mycorrhiza with arbuscules and vesicles, which was not confirmed in the original description of the species. Comparisons of E. japonicum 18S sequences with Glomeromycota DNA sequences available in a public database indicated that E. japonicum is a cosmopolitan species and is mainly associated with plants in natural habitats such as grasslands, shrublands, and forests. Phylogenetic analyses also confirmed the autonomy of E. crypticum, another known species of the genus Epigeocarpum whose sporocarps were originally found in Brazil.
“Matsushimeji” identified as Tricholoma albobrunneum belonging to sect. Genuina is an edible mushroom commonly used in Japan. This species has been suggested to include another cryptic species with common morphological characteristics and identical internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nuc rDNA). We compared T. albobrunneum specimens sampled in Japan and Denmark, including the holotype. Phylogenetic analyses of nuc rDNA ITS and intergenic spacer 1 (IGS1) regions, and tef-1a and rpb2 genes identified two phylogroups in Japanese T. albobrunneum. In addition, both Japanese phylogroups of T. albobrunneum were distinguished from European T. albobrunneum. Concatenated phylogenetic tree analysis based on these four DNA regions also distinguished two Japanese clades within T. albobrunneum. Here, we report two new species, T. matsushimeji distributed on Honshu Island under two-needle pines and T. miyama-matsushimeji on Rishiri Island under a five-needle pine from Japan.
Astraeus species are valuable edible ectomycorrhizal mushrooms, particularly in Asia. The partial hypogenous nature of Astraeus fruit bodies in soil present challenges for sample collection and studying species distribution. In this study, we developed a PCR-based approach for identifying Astraeus. Two universal primers (AUPF1/AUPR1 and AUPF3/AUPR3) and specific primers for A. asiaticus (AAF4/AAR4), A. odoratus (AOF4/AOR4), and A. sirindhorniae (ASF2/ASR4) were designed based on the alignment of internal transcribed spacer sequences from various Astraeus species. Primer verification was performed by generating amplicons from extracted DNA of Astraeus fruit bodies and 130 soil samples collected from beneath various host plants of Astraeus spp. These novel primers were efficient and precise in identifying Astraeus species. Our results have implications for multi-sample assays for Astraeus identification and investigations into species distribution through large-scale inventories.
Tricholoma matsutake is an important wild edible mycorrhizal mushroom in Japan. The domestic matsutake harvest has been declining for 80 y. Nagano Prefecture, a central region of Japan, is the area that has been most productive of matsutake during the past 20 y. To study how matsutake harvesting was affected by forest management, an experimental site was established in a pine forest in Nagano Prefecture in 1980. Two plots were established: a managed plot (MP) in which all trees except pine were cut, pine trees were thinned, and soil litter layer was removed; and a control plot (CP) with no forest management. The numbers of matsutake fruiting bodies were recorded annually in both plots for 41 y. The annual matsutake harvest was stable in the MP but decreased in the CP, while the number of shiro increased in the MP and was stable at the low level in the CP. These trends were suggested to be affected significantly by the thicker humus layer and large root biomass other than pines in the CP. This is the longest record of matsutake harvest in the managed forest, which reveals the importance of such long-term forest management for the sustainable matsutake harvesting.
The C. cinerea septins are observed at the hyphal tip and septum in the growing vegetative hyphae. In Saccharomyces cerevisiae, septin dynamics have been suggested to be regulated by septin phosphorylation through protein kinases such as Cla4. To gain insight into the relationship between C. cinerea septins and C. cinerea Cla4 protein kinase (Cc.Cla4), we conducted disruption of the C. cinerea cla4 gene (Cc.cla4) by CRISPR/Cas9 system and analyzed the phenotypes of Cc.cla4Δ strains. The vegetative hyphae of the Cc.cla4Δ strains grow slowly in the zigzag shape and are highly branched compared to the wild-type hyphae. The enhanced green fluorescent protein (EGFP) tagged Cc.Cdc3 septin in the Cc.cla4Δ strain was observed in the growing vegetative hyphae. Although EGFP-Cc.Cdc3 signals localized at the plasma membrane of the hyphal tip region, they did not accumulate as a dome with a hole at the hyphal tip as observed in the wild type. Instead, the Cc.Cdc3 was found to stay at the hyphal tip, which split into two tips thereafter. These results suggest that Cc.Cla4 is required for assembly of the hyphal tip apparatus that enables the rapid linear growth of the apical cells.
Sclerotinia borealis is among the most psychrophilic snow mold pathogen, mainly attacking Poaceae crops such as winter wheat and forage grasses as well as non-Poaceae plants under snow. Isolates were collected through the extensive surveys from North Atlantic islands through European and Asian Russia to Japan. Morphological characterization as well as the effects of temperature and water potential on mycelial growth defined three varieties, i.e., vars. borealis, subarctica, and okhotskana. Phylogenetic analyses based on ITS and EF1-α regions of genomic DNA indicated the monophyly of S. borealis and supported varietal differentiation.