Corrigendum: He XL, Horak E, Wang D, Li TH, Peng WH, Gan BC (2019) Descriptions of five new species in Entoloma subgenus Claudopus from China, with molecular phylogeny of Entoloma s.l. MycoKeys 61: 1-26. https://doi.org/10.3897/mycokeys.61.46446.

[This corrects the article DOI: 10.3897/mycokeys.61.46446.].

Descriptions of five new species in Entoloma subgenus Claudopus from China, with molecular phylogeny of Entoloma s.l.

Entoloma subgenus Claudopus is widely distributed, yet the taxonomy and systematics of its species are still poorly documented. In the present study, more than forty collections of Claudopus were gathered in China and subsequently analysed, based on morphological and molecular data. The results revealed first a high level of species diversity of Claudopus in China and second, there is a wide ecological range regarding the substrates and the habitats ranging from temperate, tropical to subalpine locations. Based on morphological and molecular evidence, five novel species from China are proposed, viz. E. conchatum, E. flabellatum, E. gregarium, E. pleurotoides and E. reductum. Molecular phylogeny of Entoloma s.l. was also reconstructed, based on 187 representatives of Entoloma s.l. by employing the combined ITS, LSU, mtSSU and RPB2 sequences. Ten monophyletic clades (Claudopus, Leptonia, Nolanea, Cuboid-spored Inocephalus, "Alboleptonia", Cyanula, Pouzarella, Rhodopolia, Prunuloides and Rusticoides) were recovered, while 13 taxa could not be placed in any defined clades. The results confirmed that Claudopus in a traditional morphological sense is not monophyletic and the Rusticoides-group, previously considered within Claudopus, formed a separate clade; but section Claudopus and relatives of E. undatum belong to a distinctive monophyletic group. Despite some monophyletic groups in Entoloma s.l. being distinctive in both morphology and molecular phylogeny, they were still treated as subgenera of Entoloma s.l. temporarily, because accepting them as genera will make Entoloma s.l. paraphyletic.

Validation of reference genes for quantitative gene expression analysis in Auricularia cornea.

Auricularia cornea Ehrenb., previously named A. polytricha (Mont.) Sacc, has become one of the most widely cultivated mushrooms in China. Considerable research has been conducted on its cultivation, pathogen identification, proteomics, and more. However, to the best of our knowledge, no studies have been performed on reference-gene validation in this species. Formerly, reference genes were selected for their expression levels only relied upon from others species, owing to the fact that the gene stability in this species is unknown. In this study, nine candidate genes, including tubulin alpha-1A chain (TUBA1A), ß-tubulin (Btu), phosphoglucomutase (Pgm), actin 1 (Act1), protein phosphatase 2A regulatory subunit (PP2A), polyubiquitin (UBQ), glyceraldehyde-3-phosphate dehydrogenase (Gapdh), 18S ribosomal protein (18S) and 28S ribosomal protein (28S), were evaluated among different strains and developmental stages. Four algorithms (i.e., geNorm, NormFinder, BestKeeper and RefFinder) were used to analyze candidate genes. The results revealed that UBQ was the most stable reference gene, while 18S was the least stable. Despite these results, the candidate genes were largely inadequate and only two were considered suitable. Based on candidate gene stability, PP2A and UBQ were identified as a set of usable interior control genes for future analyses in this species. This is the first systematic study conducted for selecting reference genes in A. cornea, and lays the foundation for identifying genes and quantifying gene expression in this species.

Three new species of EntolomasubgenusPouzarella from China based on morphological and molecular data.

In the present paper, three additional species of EntolomasubgenusPouzarella viz. E.erectoides, E.griseocarpum and E.rubropilosum are described from China. E.rubropilosum is a typical species in section Pouzarella; E.griseocarpum and E.erectoides are members of sect. Dysthales. The taxa are further confirmed by ITS, RPB2, LSU and mtSSU analyses and phylogenetic relationships with other Entolomasubgen.Pouzarella species are also discussed. ITS sequence analysis showed that the sizes of the entire ITS region and ITS1 are remarkably divergent, while the ITS2 is conserved in length within Entolomasubgen.Pouzarella. Molecular analyses, based on the combined dataset, demonstrated that species diversity of subgen.Pouzarella in China is much higher than previously thought, in the present study twenty phylogenetic species from China are taken into consideration. On the other hand, morphological and molecular analyses suggested that classification of Entolomasubgen.Pouzarella probably has to be fundamentally re-adjusted based on additional data.

Intra- and inter-isolate variation of ribosomal and protein-coding genes in Pleurotus: implications for molecular identification and phylogeny on fungal groups.

BACKGROUND: The internal transcribed spacer (ITS), RNA polymerase II second largest subunit (RPB2), and elongation factor 1-alpha (EF1α) are often used in fungal taxonomy and phylogenetic analysis. As we know, an ideal molecular marker used in molecular identification and phylogenetic studies is homogeneous within species, and interspecific variation exceeds intraspecific variation. However, during our process of performing ITS, RPB2, and EF1α sequencing on the Pleurotus spp., we found that intra-isolate sequence polymorphism might be present in these genes because direct sequencing of PCR products failed in some isolates. Therefore, we detected intra- and inter-isolate variation of the three genes in Pleurotus by polymerase chain reaction amplification and cloning in this study. RESULTS: Results showed that intra-isolate variation of ITS was not uncommon but the polymorphic level in each isolate was relatively low in Pleurotus; intra-isolate variations of EF1α and RPB2 sequences were present in an unexpectedly high amount. The polymorphism level differed significantly between ITS, RPB2, and EF1α in the same individual, and the intra-isolate heterogeneity level of each gene varied between isolates within the same species. Intra-isolate and intraspecific variation of ITS in the tested isolates was less than interspecific variation, and intra-isolate and intraspecific variation of RPB2 was probably equal with interspecific divergence. Meanwhile, intra-isolate and intraspecific variation of EF1α could exceed interspecific divergence. These findings suggested that RPB2 and EF1α are not desirable barcoding candidates for Pleurotus. We also discussed the reason why rDNA and protein-coding genes showed variants within a single isolate in Pleurotus, but must be addressed in further research. CONCLUSIONS: Our study demonstrated that intra-isolate variation of ribosomal and protein-coding genes are likely widespread in fungi. This has implications for studies on fungal evolution, taxonomy, phylogenetics, and population genetics. More extensive sampling of these genes and other candidates will be required to ensure reliability as phylogenetic markers and DNA barcodes.

White mold on cultivated morels caused by Paecilomyces penicillatus.

Morchella (morel) includes prized edible and medical mushrooms in the world. Since 2012, commercial cultivation of morels in the field has developed rapidly in China. However, coupled with the rapid expansion of morel cultivation, diseases have been become serious threats to morel production. White mold is one of the most serious diseases on cultivated morels. This study aimed to confirm this pathogen by following Koch's postulates, and to identify it using molecular evidence. Our results indicated that healthy Morchella fruiting bodies inoculated with Paecilomyces sp. isolates produced typical white mold symptoms, and the internal transcribed spacer sequences of the Paecilomyces sp. were 99% similar to that recovered from an epitype of Paecilomyces penicillatus. Therefore, P. penicillatus was considered to be the causative agent of white mold. White mold occurred from the initial harvest to the storage and preservation process, and it produced white mold-like symptoms on the caps and stripes of Morchella. This is the first time that white mold has been reported on cultivated Morchella.

Phylogenetic relationship of two popular edible Pleurotus in China, Bailinggu (P. eryngii var. tuoliensis) and Xingbaogu (P. eryngii), determined by ITS, RPB2 and EF1α sequences.

The aims of this study are to assess the utility of the internal transcribed spacer (ITS) region, and partial translation elongation factor (EF1α) and RNA polymerase II (RPB2) genes, for differentiation of Bailinggu, P. eryngii, and P. nebrodensis; to reconstruct phylogenetic relationships between the three species; and to confirm the taxonomic status of Bailinggu based on ribosomal and protein-coding genes. Pairwise genetic distances between Bailinggu, P. eryngii, and related Pleurotus strains were calculated by using the p-distance model, and molecular phylogeny of these isolates was estimated based on ITS, RPB2, and EF1α using maximum parsimony and Bayesian methods. Differences in ITS, RPB2, and EF1α sequences show that Bailinggu, P. eryngii, and P. nebrodensis are distinct at the species level. Phylogenetic analyses reveal that P. eryngii is closer to P. nebrodensis than to Bailinggu. Sequence analyses of ribosomal and protein-coding genes confirm that P. eryngii var. tuoliensis is identical to Bailinggu. P. eryngii var. tuoliensis should be raised to species level or a new name should be introduced for Bailinggu after a thorough investigation into Pleurotus isolates from Ferula in Xinjiang Province. This study helps to resolve uncertainty regarding Bailinggu, P. eryngii and P. nebrodensis, improving the resource management of these strains. ITS, EF1α, and RPB2 sequences can be used to distinguish Bailinggu, P. eryngii and P. nebrodensis as three different species, and P. eryngii var. tuoliensis should be the scientific name for Bailinggu at present.

[Study on methods of deproteinization from Phellinus baumii polysaccharide].

OBJECTIVE: To study and compare different methods and influencing factors of deproteinization from Phellinus baumii polysaccharide. METHODS: Sevag method and TCA method were used to remove protein from crude polysaccharide of Phellinus baumii, respectively. RESULTS: The effect of TCA method on removing protein was superior to that of Sevag method,and the optimum conditions of removing protein with TCA were as follows: 30 min treating time, 5% trichloroacetic acid,3 times treatemt. Under these conditions the rate of protein-removed was 82% and the rate of polysaccharide-lost was 10.8%. CONCLUSION: TCA method is the optimal mean for deproteinization from Phellinus baumii polysaccharide.