Pleurotus tuber-regium is a sclerotium-producing mushroom with pharmacological and nutritional value. Many researches applied molecular biological strategies that allow analyzing numerous genes expressed in response to different growth stresses or tissues. In order to capture accurate and reliable results in gene expression studies, it is necessary to select appropriate internal reference genes. They must have relatively uniform expression in the majority of tested samples, thereby reducing the impact of the sample itself on the RT-qPCR results. The selection and validation of 14 candidate reference genes, including α-tub, β-tub, γ-tub, GAPDH, Tif-5a, Tef-1α, ATPase, acyl-CoA, U2ribo, S/G, Wlp, VtpAsE, E3upl, and Sdh, were carried out for gene expression analysis in P. tuber-regium mycelia in response to different temperatures, drought levels, and salinity shifts, fruitbody, and sclerotium. Four statistical algorithms, NormFinder, geNorm, BestKeeper, and RefFinder, were recruited to evaluate the transcription stability of candidate reference genes. The RT-qPCR expression stability analysis indicated that the E3upl and Tif-5a were the most stable expressed genes among all experimental samples, so the combination of these two reference genes is suitable for the analysis of gene expression patterns in P. tuber-regium.
In this study, we identified Sec5 in Bipolaris maydis, a homologue of Sec5 in Saccharomyces cerevisiae and a possible exocyst component of the fungus. To examine how Sec5 affects the life cycle of B. maydis, we generated null mutant strains of the gene (Δsec5). The Δsec5 strains showed a strong reduction in hyphal growth and a slight reduction in pathogenicity. In sexual reproduction, they possessed the ability to develop pseudothecia. However, all ascospores were aborted in any of the asci obtained from crosses between Δsec5 and the wild-type. Our cytological study revealed that the abortion was caused by impairments of the post-meiotic stages in ascospore development, where ascospore delimitation and young spore elongation occur.
Morphological characters and multi-gene phylogenetic analyses were used to identify Retiboletus specimens collected in northern Thailand. Retiboletus brevibasidiatus is described as new to science, whereas R. fuscus and R. nigrogriseus are reported for the first time from Thailand. Retiboletus brevibasidiatus produces medium-sized basidiomes, with a dark blonde to clay pileus and densely reticulate stipe mostly on the upper part with pale yellow to chrome yellow basal mycelium. It is difficult to separate R. brevibasidiatus from other closely related species on the basis of macroscopic characters. However, the new species can be distinguished by microscopic characters, mostly the shorter basidia. The macro- and micro-morphology of the R. fuscus and R. nigrogriseus collections from Thailand fit well with the previous descriptions of materials from China and Japan. Detailed descriptions, molecular phylogeny, and illustrations of the three species are provided.
“Kakishimeji” identified as Tricholoma ustale and belonging to Tricholoma sect. Genuina is a common poisonous mushroom in Japan. Kakishimeji contains the toxic compound ustalic acid and causes digestive trouble. However, this fungus is consumed in some regions of Japan without any digestive issues. We clarified the probable species complex of Kakishimeji based on a phylogenetic analysis. We collected 89 basidioma specimens of Kakishimeji and related species from various forest sites in Japan and conducted phylogenetic analyses using 7 nuclear and mitochondrial gene sequences. Kakishimeji was found to consist of four distinct phylogenetic clades based on all DNA regions tested. Of these, two clades included European T. stans and T. albobrunneum type specimens. Another two clades consisted of sister clades to T. pessundatum and T. ustaloides. In addition, all four phylogenetic clades of Kakishimeji had different spore and basidium sizes. Therefore, we regarded the latter two clades as two new Tricholoma species: T. kakishimeji and T. kakishimejioides.
Truffle cultivation is successfully applied throughout the world for several truffles of European origin. However, just two Chinese black truffles (Tuber indicum and T. himalayense) have been cultivated with a favorable outcome so far. Tuber pseudohimalayense is a black truffle of significant economic relevance in China, but little is known about its mycorrhizal ecology and it is not cultivated in orchards yet. Here, we selected seven broad-leaved tree species (Quercus fabrei, Q. aliena, Castanea mollissima, Carya illinoinensis, Q. glauca, Castanopsis orthacantha, Betula costata), and one coniferous tree (Pinus armandii), and inoculated them with T. pseudohimalayense spore suspension using axenically germinated seedlings under greenhouse conditions. The obtained mycorrhizae, well-developed, were analyzed from the morpho-anatomical and molecular points of view, and their main characteristics described. Synthesized T. pseudohimalayense mycorrhizae showed similar characters on all tree species, with a typical interlocking pseudoparenchymatous mantle and Hartig net, swollen appearance, yellow-brownish color, and long hyaline emanating hyphae with right-angle ramifications. These features are similar to those reported for mycorrhizae formed by related black truffle species. The successful mycorrhizal synthesis of T. pseudohimalayense on multiple trees species indicates that it has potential for cultivation in China.
Absidia healeyae is a new species described in the Mucorales genus Absidia after screening 16 strains of Absidia isolated from seven locations in the state of Victoria in Australia. After initial analysis of the large ribosomal subunit sequence, the genomes of representative strains from two clades were sequenced using short paired-reads. Additional taxonomic markers extracted from the genome sequencing data support the novelty of A. healeyae. The identification of a new species in the genus Absidia, from a relatively small collection of isolates, hints at an unexplored diversity in the early diverging lineages of fungi in Australia.
Hemileia gardeniae-floridae is an accepted name for a Cape jasmine (Gardenia jasminoides) rust fungus distributed in East Asia. The fungus name was based on uredinial anamorph collected in Taiwan in 1931. The fungus was rarely collected in Taiwan and southern Japan, and its telial stage remained unknown. Microscopic examination of the type materials of H. gardeniae-floridae and Uredo gardeniae-floridae, which was once proposed to replace H. gardeniae-floridae, resulted in discovery of teliospores on the type of U. gardeniae-floridae. The teliospores are mostly napiform and produced on a sporogenous cell emerging through host stoma. A hemileioid rust fungus, producing both urediniospores and teliospores on Golden gardenia (G. sootepensis), was found in Thailand and morphologically identified to H. gardeniae-floridae. Another Hemileia species on Forest jasmine (G. thunbergia), H. gardeniae-thunbergiae, first found in Angola, Africa, is distinct from H. gardeniae-floridae in producing smaller urediniospores than those of H. gardeniae-floridae.
In order to elucidate the lifecycle of Coprinus comatus, we examined the number of nuclei in basidiospores, hyphal cells and oidia. Basidiospores isolated from the fruiting bodies of four Japanese strains were binucleate. In both primary and secondary mycelia, most of the cells were binucleate. In addition, oidia and oidiophores were observed for the first time in this mushroom and most of the oidia were binucleate. Based on these results, the lifecycle of C. comatus was inferred to be as follows. A homokaryotic binucleate basidiospore germinates and produces homokaryotic binucleate hyphae. After mating between compatible homokaryotic binucleate hyphae, a heterokaryotic binucleate secondary mycelium is produced. If environmental conditions are suitable for fruiting, homokaryotic binucleate basidiospores in the fruiting body are produced.