Exploring structural and molecular diversity of Ericaceae hair root mycobionts: a comparison between Northern Bohemia and Argentine Patagonia.

Core Ericaceae produce delicate hair roots with inflated rhizodermal cells that host plethora of fungal symbionts. These poorly known mycobionts include various endophytes, parasites, saprobes, and the ericoid mycorrhizal (ErM) fungi (ErMF) that form the ErM symbiosis crucial for the fitness of their hosts. Using microscopy and high-throughput sequencing, we investigated their structural and molecular diversity in 14 different host × site combinations in Northern Bohemia (Central Europe) and Argentine Patagonia (South America). While we found typical ericoid mycorrhiza in all combinations, we did not detect ectomycorrhiza and arbuscular mycorrhiza. Superficial mantles of various thickness formed by non-clamped hyphae were observed in all combinations except Calluna vulgaris from N. Bohemia. Some samples contained frequent intercellular hyphae while others possessed previously unreported intracellular haustoria-like structures linked with intracellular hyphal coils. The 711 detected fungal OTU were dominated by Ascomycota (563) and Basidiomycota (119), followed by four other phyla. Ascomycetes comprised Helotiales (255), Pleosporales (53), Chaetothyriales (42), and other 19 orders, while basidiomycetes Sebacinales (42), Agaricales (28), Auriculariales (7), and other 14 orders. While many dominant OTU from both hemispheres lacked close relatives in reference databases, many were very similar to identical to unnamed sequences from around the world. On the other hand, several significant ericaceous mycobionts were absent in our dataset, incl. Cairneyella, Gamarada, Kurtia, Lachnum, and Leohumicola. Most of the detected OTU could not be reliably linked to a particular trophic mode, and only two could be reliably assigned to the archetypal ErMF Hyaloscypha hepaticicola. Probable ErMF comprised Hyaloscypha variabilis and Oidiodendron maius, both detected only in N. Bohemia. Possible ErMF comprised sebacinoid fungi and several unnamed members of Hyaloscypha s. str. While H. hepaticicola was dominant only in C. vulgaris, this model ErM host lacked O. maius and sebacinoid mycobionts. Hyaloscypha hepaticicola was absent in two and very rare in six combinations from Patagonia. Nine OTU represented dark septate endophytes from the Phialocephala fortinii s. lat.-Acephala applanata species complex, including the most abundant OTU (the only detected in all combinations). Statistical analyses revealed marked differences between N. Bohemia and Patagonia, but also within Patagonia, due to the unique community detected in a Valdivian temperate rainforest. Our results show that the ericaceous hair roots may host diverse mycobionts with mostly unknown functions and indicate that many novel ErMF lineages await discovery. Transhemispheric differences (thousands of km) in their communities may be evenly matched by local differences (scales of km, m, and less).

A review of Hyphodiscaceae.

In a recently published classification scheme for Leotiomycetes, the new family Hyphodiscaceae was erected; unfortunately, this study was rife with phylogenetic misinterpretations and hampered by a poor understanding of this group of fungi. This manifested in the form of an undiagnostic familial description, an erroneous familial circumscription, and the redescription of the type species of an included genus as a new species in a different genus. The present work corrects these errors by incorporating new molecular data from this group into phylogenetic analyses and examining the morphological features of the included taxa. An emended description of Hyphodiscaceae is provided, notes and descriptions of the included genera are supplied, and keys to genera and species in Hyphodiscaceae are supplied. Microscypha cajaniensis is combined in Hyphodiscus, and Scolecolachnum nigricans is a taxonomic synonym of Fuscolachnum pteridis. Future work in this family should focus on increasing phylogenetic sampling outside of Eurasia and better characterising described species to help resolve outstanding issues. Citation: Quijada L, Baral HO, Johnston PR, Pärtel K, Mitchell JK, Hosoya T, Madrid H, Kosonen T, Helleman S, Rubio E, Stöckli E, Huhtinen S, Pfister DH (2022). A review of Hyphodiscaceae. Studies in Mycology 103: 59-85. doi: 10.3114/sim.2022.103.03.

First Report of Collar Rot Caused by Sclerotinia sclerotiorum on Sesame (Sesamum indicum) in Mexico.

Sesame (Sesamum indicum L.: Pedaliaceae) is the second most cultivated oilseed in Mexico with 80,000 ha per year. The seeds of this crop are used as a condiment, for the extraction of oil, and its medicinal properties. In October 2020, collar rot symptoms were observed in six sesame fields (SOPC-9539 TD variety) located in the Carrizo Valley (26°15'33.1"N; 109°01'37.9"W), El Fuerte, Sinaloa, México. Initially, small brown spots in the basal stem of the infected plants were observed. At advanced stages of the disease, the circumference of stem was necrotic with the presence of white mycelium that extends to the roots. Infected plants were showing symptoms of yellowing, wilting, and finally death. Disease incidence was estimated at 15%, counting the total of diseased plants in five counts done in arbitrary quadrants within the sesame fields. For fungal isolation, stem sections from the symptomatic basal stem were surface disinfected with 1.5% sodium hypochlorite for 2 min, then triple rinsed with sterile distilled water. The tissue sections were dried on sterile blotting paper and plated in Petri dishes with potato dextrose agar (PDA) culture medium. The plates were incubated at 28ºC in darkness for 48 h. Sclerotinia-like colonies were consistently isolated and four isolates from different locations were purified by the hyphal-tip method. Fungal colonies were formed of compact white mycelium, with the formation of sclerotia on the margin of the plate 6 days after inoculating PDA cultures. Sclerotia averaged 3.1 mm in diameter and 0.024 g. One isolate was deposited in the Culture Collection of Phytopathogenic Fungi of the Faculty of Agriculture of Fuerte Valley at the Sinaloa Autonomous University under Accession no. FAVF654. To confirm identification, genomic DNA was extracted from one isolate, and the internal transcribed spacer (ITS) region was amplified by PCR and sequenced directly using the primer pair ITS5/ITS4 (White et al. 1990). The resulting consensus sequence was deposited in GenBank under accession no. ON401416. BLASTn alignments in GenBank showed 100% identity of our sequence with the sequence of the type strain of Sclerotinia sclerotiorum ATCC 46762 (accession no. JX648201). Pathogenicity of the fungus was demonstrated by inoculating healthy sesame plants (Dormilón and SOPC-9539 TD ies), germinated in plastic pots with sterile substrate. Plants were inoculated with the FAVF654 isolate by applying 3 sclerotia at the base of each of the 12 plants. Twelve plants were left uninoculated, which served as controls. All the inoculated plants, of both varieties, developed the characteristic symptoms of the disease 7 days after inoculation, while the control plants remained symptomless. The pathogenicity test was performed twice with the same result. The fungus was reisolated from all the inoculated plants, thus fulfilling Koch's postulates. Sclerotinia sclerotiorum has been reported on sesame plants in Bulgaria and Korea (Farr and Rossman, 2022). To our knowledge, this is the first report of Sclerotinia sclerotiorum causing collar rot in sesame plants in Mexico and the Americas. This disease considerably reduces the yield of sesame; therefore it is necessary to develop effective disease-management strategies.

Taxonomy and systematics of Hyaloscyphaceae and Arachnopezizaceae.

The circumscription and composition of the Hyaloscyphaceae are controversial and based on poorly sampled or unsupported phylogenies. The generic limits within the hyaloscyphoid fungi are also very poorly understood. To address this issue, a robust five-gene Bayesian phylogeny (LSU, RPB1, RPB2, TEF-1α, mtSSU; 5521 bp) with a focus on the core group of Hyaloscyphaceae and Arachnopezizaceae is presented here, with comparative morphological and histochemical characters. A wide representative sampling of Hyaloscypha supports it as monophyletic and shows H. aureliella (subgenus Eupezizella) to be a strongly supported sister taxon. Reinforced by distinguishing morphological features, Eupezizella is here recognised as a separate genus, comprising E. aureliella, E. britannica, E. roseoguttata and E. nipponica (previously treated in Hyaloscypha). In a sister group to the Hyaloscypha-Eupezizella clade a new genus, Mimicoscypha, is created for three seldom collected and poorly understood species, M. lacrimiformis, M. mimica (nom. nov.) and M. paludosa, previously treated in Phialina, Hyaloscypha and Eriopezia, respectively. The Arachnopezizaceae is polyphyletic, because Arachnoscypha forms a monophyletic group with Polydesmia pruinosa, distant to Arachnopeziza and Eriopezia; in addition, Arachnopeziza variepilosa represents an early diverging lineage in Hyaloscyphaceae s.str. The hyphae originating from the base of the apothecia in Arachnoscypha are considered anchoring hyphae (vs a subiculum) and Arachnoscypha is excluded from Arachnopezizaceae. A new genus, Resinoscypha, is established to accommodate Arachnopeziza variepilosa and A. monoseptata, originally described in Protounguicularia. Mimicoscypha and Resinoscypha are distinguished among hyaloscyphoid fungi by long tapering multiseptate hairs that are not dextrinoid or glassy, in combination with ectal excipulum cells with deep amyloid nodules. Unique to Resinoscypha is cyanophilous resinous content in the hairs concentrated at the apex and septa. Small intensely amyloid nodules in the hairs are furthermore characteristic for Resinoscypha and Eupezizella. To elucidate species limits and diversity in Arachnopeziza, mainly from Northern Europe, we applied genealogical concordance phylogenetic species recognition (GCPSR) using analyses of individual datasets (ITS, LSU, RPB1, RPB2, TEF-1α) and comparative morphology. Eight species were identified as highly supported and reciprocally monophyletic. Four of these are newly discovered species, with two formally described here, viz. A. estonica and A. ptilidiophila. In addition, Belonium sphagnisedum, which completely lacks prominent hairs, is here combined in Arachnopeziza, widening the concept of the genus. Numerous publicly available sequences named A. aurata represent A. delicatula and the confusion between these two species is clarified. An additional four singletons are considered to be distinct species, because they were genetically divergent from their sisters. A highly supported five-gene phylogeny of Arachnopezizaceae identified four major clades in Arachnopeziza, with Eriopezia as a sister group. Two of the clades include species with a strong connection to bryophytes; the third clade includes species growing on bulky woody substrates and with pigmented exudates on the hairs; and the fourth clade species with hyaline exudates growing on both bryophytes and hardwood. A morphological account is given of the composition of Hyaloscyphaceae and Arachnopezizaceae, including new observations on vital and histochemical characters. Citation: Kosonen T, Huhtinen S, Hansen K. 2021. Taxonomy and systematics of Hyaloscyphaceae and Arachnopezizaceae. Persoonia 46: 26-62. https://doi.org/10.3767/persoonia.2021.46.02.

Root-associated fungal microbiota of nonmycorrhizal Arabis alpina and its contribution to plant phosphorus nutrition.

Most land plants live in association with arbuscular mycorrhizal (AM) fungi and rely on this symbiosis to scavenge phosphorus (P) from soil. The ability to establish this partnership has been lost in some plant lineages like the Brassicaceae, which raises the question of what alternative nutrition strategies such plants have to grow in P-impoverished soils. To understand the contribution of plant-microbiota interactions, we studied the root-associated fungal microbiome of Arabis alpina (Brassicaceae) with the hypothesis that some of its components can promote plant P acquisition. Using amplicon sequencing of the fungal internal transcribed spacer 2, we studied the root and rhizosphere fungal communities of A. alpina growing under natural and controlled conditions including low-P soils and identified a set of 15 fungal taxa consistently detected in its roots. This cohort included a Helotiales taxon exhibiting high abundance in roots of wild A. alpina growing in an extremely P-limited soil. Consequently, we isolated and subsequently reintroduced a specimen from this taxon into its native P-poor soil in which it improved plant growth and P uptake. The fungus exhibited mycorrhiza-like traits including colonization of the root endosphere and P transfer to the plant. Genome analysis revealed a link between its endophytic lifestyle and the expansion of its repertoire of carbohydrate-active enzymes. We report the discovery of a plant-fungus interaction facilitating the growth of a nonmycorrhizal plant under native P-limited conditions, thus uncovering a previously underestimated role of root fungal microbiota in P cycling.

Plant-mediated effects of soil phosphorus on the root-associated fungal microbiota in Arabidopsis thaliana.

Plants respond to phosphorus (P) limitation through an array of morphological, physiological and metabolic changes which are part of the phosphate (Pi) starvation response (PSR). This response influences the establishment of the arbuscular mycorrhizal (AM) symbiosis in most land plants. It is, however, unknown to what extent available P and the PSR redefine plant interactions with the fungal microbiota in soil. Using amplicon sequencing of the fungal taxonomic marker ITS2, we examined the changes in root-associated fungal communities in the AM nonhost species Arabidopsis thaliana in response to soil amendment with P and to genetic perturbations in the plant PSR. We observed robust shifts in root-associated fungal communities of P-replete plants in comparison with their P-deprived counterparts, while bulk soil communities remained unaltered. Moreover, plants carrying mutations in the phosphate signaling network genes, phr1, phl1 and pho2, exhibited similarly altered root fungal communities characterized by the depletion of the chytridiomycete taxon Olpidium brassicae specifically under P-replete conditions. This study highlights the nutritional status and the underlying nutrient signaling network of an AM nonhost plant as previously unrecognized factors influencing the assembly of the plant fungal microbiota in response to P in nonsterile soil.

Gamarada debralockiae gen. nov. sp. nov.-the genome of the most widespread Australian ericoid mycorrhizal fungus.

This study describes a novel ericoid mycorrhizal fungus (ErMF), Gamarada debralockiae Midgley and Tran-Dinh gen. nov. sp. nov. Additionally, catabolism was explored from a genomic perspective. The nuclear and mitochondrial genomes of G. debralockiae were sequenced. Morphological characteristics were assessed on various media. Catabolic genes of G. debralockiae were explored using SignalP and dbCAN. Phylogenetic comparisons were undertaken using Phylogeny.fr. The 58.5-Mbp draft genome of G. debralockiae contained 17,075 putative genes. The complete mitochondrial genome was 28,168 bp in length. In culture, G. debralockiae produces slow-growing non-sporulating colonies. Gamarada debralockiae has many putative secreted catabolic enzymes. Phylogeny indicated G. debralockiae was distinct from known ascomycetous ErMF: Pezoloma ericae, Meliniomyces spp., Oidiodendron spp., and Cairneyella variabilis. It is closely related to many undescribed plant root-associated fungi and its nearest described relative is Hyphodiscus brevicollaris. Gamarada debralockiae has been recovered from virtually all Australian ericoid mycorrhizal studies and biogeographic data suggests the taxon is widespread in Australia. Gamarada debralockiae has similar catabolic potential to C. variabilis and co-occurs with C. variabilis at Australian sites. Plants that host multiple ErMF may benefit from subtle differences in catabolism that improve access to nitrogen and phosphorus from within recalcitrant organic matter.

Localization of helotialean fungi on ectomycorrhizae of Castanopsis cuspidata visualized by in situ hybridization.

Non-ectomycorrhizal fungi that associate with typical ectomycorrhizae often remain hidden, and their localization inside ectomycorrhizal (ECM) roots has remained uncharacterized. In this study, the fungal community associated with the ectomycorrhizae of Castanopsis cuspidata was investigated using a culture-dependent isolation technique. Additionally, the species composition and localization were determined using molecular techniques. The results of the isolation and identification of fungal species revealed the predominance of a few species belonging to the order Helotiales. Furthermore, the fungal community structures were significantly different depending on the taxa of the ectomycorrhiza-forming fungi. A taxon-specific probe was developed to analyze the localization of one dominant Hyaloscyphaceae (Helotiales) species in ECM tissues by in situ hybridization. Hybridization signals were detected on the surface of the fungal mantle and around the ECM fungal cells within the mantle. Hyphal penetration into ECM hyphal cells of fungal mantles was also observed. Signals were not detected in the Hartig net or plant tissues inside the mantle in healthy ectomycorrhizae. These findings suggest that the analyzed species interact not only with host plant as root endophyte but also directly with the ECM fungi.

Sexual and asexual states of some endophytic Phialocephala species of Picea.

Unidentified DNA sequences in isolation-based or culture-free studies of conifer endophytes are a persistent problem that requires a field approach to resolve. An investigation of foliar endophytes of Picea glauca, P. mariana, P. rubens and Pinus strobus in eastern Canada, using a combined field, morphological, cultural and DNA sequencing approach, resulted in the frequent isolation of Phialocephala spp. and the first verified discovery of their mollisia-like sexual states in the field. Phialocephala scopiformis and Ph. piceae were the most frequent species isolated as endophytes from healthy conifer needles. Corresponding Mollisia or mollisioid sexual states for Ph. scopiformis, Ph. piceae and several undescribed species in a clade containing Ph. dimorphospora were collected in the sampling area and characterized by analysis of the nuc internal transcribed spacer rDNA (ITS) and gene for the largest subunit of RNA polymerase II (RPB1) loci. Four novel species and one new combination in a clade containing Ph. dimorphospora, the type of Phialocephala, are presented, accompanied by descriptions of apothecia and previously undocumented synanamorphs. An epitype culture and corresponding reference sequences for Phialocephala dimorphospora are proposed. The resulting ITS barcodes linked with robust taxonomic species concepts are an important resource for future research on forest ecosystems and endophytes.

Genomic insights into the carbohydrate catabolism of Cairneyella variabilis gen. nov. sp. nov., the first reports from a genome of an ericoid mycorrhizal fungus from the southern hemisphere.

This paper describes a novel species of ericoid mycorrhizal fungus from Australia, Cairneyella variabilis, Midgley and Tran-Dinh, gen. nov. sp. nov. The genome of C. variabilis was sequenced and a draft genome assembled. The draft genome of C. variabilis is 52.4 Mbp in length, and to our knowledge, this is the first study to present a genome of an ericoid mycorrhizal fungus from the southern hemisphere. Using the SignalP and dbCAN bioinformatic pipelines, a study of the catabolic potential of C. variabilis was undertaken and showed genes for an array of degradative enzymes, most of which appear to be secreted from the hyphae, to access a suite of different carbon sources. Isolates of C. variabilis have been previously shown to utilise cellulose, carboxymethyl cellulose (CMC), cellobiose, xylan, pectin, starch and tannic acid for growth, and in the current study, putative enzymes for these processes were revealed. These enzymes likely play key roles in nutrient cycling and other edaphic processes in heathland environments. ITS phylogenetic analyses showed C. variabilis to be distinct from the fungi of the "Hymenoscyphus ericae aggregate".

New mutualistic fungal endophytes isolated from poplar roots display high metal tolerance.

This study aimed to isolate, identify, and characterise metal-tolerant fungi colonising poplar roots at a metal-contaminated phytoremediation site. Poplar roots were colonised by arbuscular mycorrhizal, ectomycorrhizal, and endophytic fungi, and the species were determined by ITS molecular analyses. Eight different isolates were successfully isolated into pure culture. Three isolates belonging to the Helotiales (P02, P06) and the Serendipita vermifera species (P04) were highly tolerant to metals (Cd, Zn, Pb, and Cu) compared to the mycorrhizal Hebeloma isolates. The three isolates degraded complex carbohydrates, such as xylan and cellulose, indicating that they could partially degrade root cell walls and penetrate into cells. This hypothesis was confirmed by further in vitro re-synthesis experiments, which showed that the three isolates colonised root tissues of poplar plantlets whereas two of them formed microsclerotia-like structures. Taken together, these results suggest an endophytic lifestyle of these isolates. This is the first evidence of S. vermifera as a root endophyte of poplar. A new endophytic putative species belonging to the Helotiales and closely related to Leohumicola is also reported. Interestingly, and when compared to mock-inoculated plants, both P06 and P04 isolates increased the number of root tips of inoculated poplar plantlets in vitro. Moreover, the S. vermifera P04 isolate also increased the shoot biomass. The results are discussed in relation to the potential use of endophytic strains for tree-based phytoremediation of metal-contaminated sites.

Acid protease production in fungal root endophytes.

Fungal endophytes are ubiquitous in healthy root tissue, but little is known about their ecosystem functions, including their ability to utilize organic nutrient sources such as proteins. Root-associated fungi may secrete proteases to access the carbon and mineral nutrients within proteins in the soil or in the cells of their plant host. We compared the protein utilization patterns of multiple isolates of the root endophytes Phialocephala fortinii s.l., Meliniomyces variabilis and Umbelopsis isabellina with those of two ectomycorrhizal (ECM) fungi, Hebeloma incarnatulum and Laccaria bicolor, and the wood-decay fungus Irpex lacteus at pH values of 2-9 on liquid BSA media. We also assessed protease activity using a fluorescently labeled casein assay and gelatin zymography and characterized proteases using specific protease inhibitors. I. lacteus and U. isabellina utilized protein efficiently, while the ECM fungi exhibited poor protein utilization. ECM fungi secreted metallo-proteases and had pH optima above 4, while other fungi produced aspartic proteases with lower pH optima. The ascomycetous root endophytes M. variabilis and P. fortinii exhibited intermediate levels of protein utilization and M. variabilis exhibited a very low pH optimum. Comparing proteolytic profiles between fungal root endophytes and fungi with well defined ecological roles provides insight into the ecology of these cryptic root associates.

Nitrogen deposition alters plant-fungal relationships: linking belowground dynamics to aboveground vegetation change.

Nitrogen (N) deposition rates are increasing globally due to anthropogenic activities. Plant community responses to N are often attributed to altered competitive interactions between plants, but may also be a result of microbial responses to N, particularly root-associated fungi (RAF), which are known to affect plant fitness. In response to N, Deschampsia cespitosa, a codominant plant in the alpine tundra at Niwot Ridge (CO), increases in abundance, while Geum rossii, its principal competitor, declines. Importantly, G. rossii declines with N even in the absence of its competitor. We examined whether contrasting host responses to N are associated with altered plant-fungal symbioses, and whether the effects of N are distinct from effects of altered plant competition on RAF, using 454 pyrosequencing. Host RAF communities were distinct (only 9.4% of OTUs overlapped). N increased RAF diversity in G. rossii, but decreased it in D. cespitosa. D. cespitosa RAF communities were more responsive to N than G. rossii RAF communities, perhaps indicating a flexible microbial community aids host adaptation to nutrient enrichment. Effects of removing D. cespitosa were distinct from effects of N on G. rossii RAF, and D. cespitosa presence reversed RAF diversity response to N. The most dominant G. rossii RAF order, Helotiales, was the most affected by N, declining from 83% to 60% of sequences, perhaps indicating a loss of mutualists under N enrichment. These results highlight the potential importance of belowground microbial dynamics in plant responses to N deposition.

Omnipolyphilins A and B: Chlorinated Cyclotetrapeptides and Naphtho-α-pyranones from the Plant Nematode-Derived Fungus Polyphilus sieberi.

Chemical exploration for two isolates of the recently described ascomycete species Polyphilus sieberi, derived from the eggs of the plant parasitic nematode Heterodera filipjevi, afforded the identification of many compounds that belong to various metabolite families: two previously undescribed chlorinated cyclotetrapeptides, omnipolyphilins A (1) and B (2), one new pyranonaphthoquinone, ventiloquinone P (3), a 6,6'-binaphto-α-pyranone dimer, talaroderxine D (4) in addition to nine known metabolites (5-13) were isolated from this biocontrol candidate. All isolated compounds were characterized by comprehensive 1D, 2D NMR, and HR-ESI-MS analyses. The absolute configurations of the cyclotetrapeptides were determined by a combination of advanced Marfey's method, ROE correlation aided by conformational analysis, and TDDFT-ECD calculations, while ECD calculations, Mosher's method, and experimental ECD spectra were used for ventiloquinone P (3) and talaroderxine D (4). Among the isolated compounds, talaroderxine D (4) showed potent antimicrobial activities against Bacillus subtilis and Staphylococcus aureus with MIC values of 2.1 and 8.3 µg mL-1, respectively. Additionally, promising inhibitory effects on talaroderxine D (4) against the formation of S. aureus biofilms were observed up to a concentration of 0.25 µg mL-1. Moreover, ophiocordylongiiside A (10) showed activity against the free-living nematode Caenorhabditis elegans.

Phylogeny and taxonomy of the genera of Erysiphaceae, part 5: Erysiphe (the "Microsphaera lineage" part 1).

In this contribution, we offer the fifth installment of a series focusing on the phylogeny and taxonomy of powdery mildews. This paper is the second segment evaluating the genus Erysiphe. The first treatment of Erysiphe focused on phylogenetically basal species in the "Uncinula lineage." This research presents a phylogenetic-taxonomic assessment of species that form the group previously referred to as the "Microsphaera lineage." Given the size of the group, we split the treatment of this lineage of Erysiphe species into two parts based on their phylogenetic placement. Phylogenetic trees based on ITS+28S data are supplemented by sequences of additional markers (CAM, GADPH, GS, RPB2, and TUB). Included in the analysis of the Microsphaera lineage is the "Erysiphe aquilegiae complex" (group, clade, cluster), which encompasses sequences obtained from an assemblage of Erysiphe species with insufficient resolution in rDNA analyses. Attempts have been made to resolve this group at the species level by applying a multilocus approach. A detailed discussion of the "Erysiphe aquilegiae complex" is provided. Sequences are provided for the first time for several species, particularly North American species, such as Erysiphe aggregata, E. erineophila, E. parnassiae, and E. semitosta. Ex-type sequences for Microsphaera benzoin and M. magnusii have been retrieved. Alphitomorpha penicillata, Microsphaera vanbruntiana, and M. symphoricarpi are epitypified with ex-epitype sequences. The new species Erysiphe alnicola, E. deutziana, E. cornigena, E. lentaginis, and E. sambucina are described, the new combinations E. lauracearum, E. passiflorae, and E. sambucicola are introduced, and the new name E. santali is proposed.

Bioactive Naphtho-α-Pyranones from Two Endophytic Fungi of the Genus Polyphilus.

In the course of our survey to study the metabolic potential of two species of a new helotialean genus Polyphilus, namely P. frankenii and P. sieberi, their crude extracts were obtained using different cultivation techniques, which led to the isolation and characterization of two new naphtho-α-pyranone derivatives recognized as a monomer (1) and its 6,6'-homodimer (2) together with two known diketopiperazine congeners, outovirin B (3) and (3S,6S)-3,6-dibenzylpiperazine-2,5-dione (4). The structures of isolated compounds were determined based on extensive 1D and 2D NMR and HRESIMS. The absolute configuration of new naphtho-α-pyranones was determined using a comparison of their experimental ECD spectra with those of related structural analogues. 6,6'-binaphtho-α-pyranone talaroderxine C (2) exhibited potent cytotoxic activity against different mammalian cell lines with IC50 values in the low micromolar to nanomolar range. In addition, talaroderxine C unveiled stronger antimicrobial activity against Bacillus subtilis rather than Staphylococcus aureus with MIC values of 0.52 µg mL-1 (0.83 µM) compared to 66.6 µg mL-1 (105.70 µM), respectively.

Phylogeny and taxonomy of the genera of Erysiphaceae, part 3: Cystotheca.

This contribution is part of a series devoted to the phylogeny and taxonomy of powdery mildews, with an emphasis on North American taxa. An overview of Cystotheca species is given, including references to ex-type sequences or, if unavailable, proposals for representative reference sequences for phylogenetic-taxonomic purposes. The new species C. mexicana is described, based on Mexican collections on Quercus glaucoides × Quercus microphylla and Quercus liebmannii × Q. microphylla. Cystotheca lanestris is reported for the first time worldwide on Quercus laceyi (Collected in Mexico) and on Q. toumeyi (collected in Arizona, USA). Cystotheca lanestris on Q. agrifolia and on Q. cerris is reported for the first time in Mexico. Epitypes with ex-epitype sequences are designated for Cystotheca wrightii, Lanomyces tjibodensis (= C. tjibodensis), Sphaerotheca kusanoi, and S. lanestris (C. lanestris).

Phylogeny and taxonomy of the genera of Erysiphaceae, part 4: Erysiphe (the "Uncinula lineage").

This is the fourth contribution within an ongoing series dedicated to the phylogeny and taxonomy of powdery mildews. This particular installment undertakes a comprehensive evaluation of a group previously referred to as the "Uncinula lineage" within Erysiphe. The genus Erysiphe is too large to be assessed in a single paper; thus, the treatment of Erysiphe is split into three parts, according to phylogenetic lineages. The first paper, presented here, discusses the most basal lineage of Erysiphe and its relationship to allied basal genera within tribe Erysipheae (i.e., Brasiliomyces and Salmonomyces). ITS+28S analyses are insufficient to resolve the basal assemblage of taxa within the Erysipheae. Therefore, phylogenetic multilocus examinations have been carried out to better understand the evolution of these taxa. The results of our analyses favor maintaining Brasiliomyces, Bulbomicroidium, and Salmonomyces as separate genera, at least for the interim, until further phylogenetic multilocus data are available for additional basal taxa within the Erysipheae. The current analyses also confirmed previous results that showed that the "Uncinula lineage" is not exclusively composed of Erysiphe species of sect. Uncinula but also includes some species that morphologically align with sect. Erysiphe, as well as species that had previously been assigned to Californiomyces and Typhulochaeta. Numerous sequences of Erysiphe species from the "Uncinula lineage" have been included in the present phylogenetic analyses and were confirmed by their position in well-supported species clades. Several species have been sequenced for the first time, including Erysiphe clintonii, E. couchii, E. geniculata, E. macrospora, and E. parvula. Ex-type sequences are provided for 16 taxa including E. nothofagi, E. trinae, and E. variabilis. Epitypes are designated and ex-epitype sequences are added for 18 taxa including Erysiphe carpophila, E. densa, and U. geniculata var. carpinicola. The new species Erysiphe canariensis is described, and the new names E. hosagoudarii and E. pseudoprunastri and the new combination E. ampelopsidis are introduced.

Mitrula aurea sp. nov., A New Aero-Aquatic Species from the Republic of Korea.

The genus Mitrula (Mitrulaceae, Helotiales), as also known as swamp beacons, inhabits submerged, decaying vegetation in standing or decaying needles, twigs, leaves, and shallow water. They play an important role in carbon cycling in some freshwater ecosystems. In the herbarium of the Korea National Arboretum (KH), seven Mitrula specimens were collected during mushroom forays in the period from 2019 to 2021. The Korean collections were found to be macromorphologically closely related to M. paludosa and M. elegans, but micromorphologically they could be distinguished by characteristics of slightly narrower asci and aseptate ascospores. Our molecular phylogenetic analyses of the internal transcribed spacer (ITS) and 28S rDNA regions also revealed that our specimens were related to M. paludosa and M. elegans, but formed a distinct clade. Based on these results, we reported our specimens as new to science and discussed the phylogeny and diversity of Mitrula species.

Phylogeny and taxonomy of the genera of Erysiphaceae, part 1: Golovinomyces.

Powdery mildews are a monophyletic group of obligate plant pathogenic fungi in the family Erysiphaceae. Powdery mildews are economically important in that they cause damage to many agriculturally significant crops and plants in ecologically important habitats. In this contribution, we introduce a new series of publications focusing on the phylogeny and taxonomy of this group, with an emphasis on specimens collected from North America. The first part of the series focuses on the genus Golovinomyces and includes a section detailing the powdery mildew species concept. We conducted analyses of Golovinomyces spp. with available rDNA sequence data from GenBank and supplemented the data set with rDNA (ITS, 28S, IGS) as well as protein-coding (GAPDH) data from 94 North American collections. Many of the species evaluated are included in phylogenetic and morphological analyses for the first time, including the American species G. americanus, G. brunneopunctatus, G. californicus, G. greeneanus, G. hydrophyllacearum, and G. sparsus. A special emphasis was placed on acquiring ex-type or ex-epitype sequences or presenting reference sequences for phylogenetic-taxonomic purposes. Three new species, G. eurybiarum, G. galiorum, and G. malvacearum, are described, and the new combinations G. fuegianus, G. mutisiae, and G. reginae are introduced. Ex-holotype sequences of Erysiphe sparsa (≡ G. sparsus) reveal that it should be reduced to synonymy with G. ambrosiae, and ex-epitype sequences of G. valerianae reveal that it should be reduced to synonymy with G. orontii. Multiple epitypes are designated with ex-epitype sequences.