Fellhaneropsis sigmoidea sp. nov. (Byssolomataceae, lichenized Ascomycota), a new foliicolous species from China, with a world key to the species of the genus Fellhaneropsis

Stictidaceae comprises taxa with diverse lifestyles. Many species in this family are drought resistant and important for studying fungal adaptation and evolution. Stictidaceae comprises 32 genera, but many of them have been neglected for decades due to the lack of field collections and molecular data. In this study, we introduce a new species Fitzroyomyces hyaloseptisporus and a new combination Fitzroyomyces pandanicola. We also provide additional morphological and molecular data for Ostropomyces pruinosellus and O. thailandicus based on new collections isolated from an unidentified woody dicotyledonous host in Chiang Rai, Thailand. Taxonomic conclusions are made with the aid of morphological evidence and phylogenetic analysis of combined LSU, ITS and mtSSU sequence data. Characteristics such as the shape and septation of ascospores and conidia as well as lifestyles among genera of Stictidaceae are discussed.

The present study aimed at elucidating the structure of Inocybe subg. Inosperma sect. Rimosae but included also representatives from subg. Mallocybe and the genus Auritella. Phylogenetic relationships were inferred using ITS, LSU and mtSSU sequence data. The analyses recovered the ingroup as a monophyletic, strongly supported clade. The results indicate that recognizing Auritella on the genus level renders Inocybe paraphyletic. The species traditionally placed in sect. Rimosae were found to be distributed over two strongly supported clades, Maculata and Rimosae s.s. The Maculata clade clusters with sect. Cervicolores and the two represent subg. Inosperma in a strict sense. Rimosae s.s. emerges as an independent, supported clade well separated from Inosperma s.s. Twenty-one terminal groups were correlated with morphologically distinct species. In addition several taxa on single branches and minor less supported clades were recovered. A key to the identified species of the Maculata and Rimosae s.s. clades which occur in Northwest Europe is provided.

Jasminum sambac L. is a species of jasmine native to a small region in the eastern Himalayas and is cultivated worldwide as an ornamental plant (USDA-ARS 2016). In Pakistan, it is cultivated for ornamental purposes throughout the country. The flowers of this plant are traditionally used in the preparation of essential oils and for making jasmine tea. The flowers and leaves also have been used in folk medicine to treat breast cancer, epilepsy, ulcers and promote wound healing (Al-Snafi 2018). In December, 2017, almost 10 leaves of 3 plants of J. sambac growing plant nursery of Gehlan, Pattoki, Punjab a province of Pakistan were observed with leaf spot disease. Infected leaves exhibited circular to sub-circular spots with indistinct margins and grey papery centers delimited by dark brown rims. For further microscopic study, the infected leaves were examined under a stereomicroscope. For the isolation and cultural studies of infecting fungus, infected parts of leaves were surface sterilized in 1% NaOCl for about 10 seconds, washed twice in sterilized distilled water, plated on potato dextrose agar (PDA) medium and incubated at 25°C for 4 days. Pure cultures were obtained having colonies of light to dark brown color. Conidia (n=20) were light brown to pale olivaceous brown, smooth, obclavate to cylindrical in shape, 99.5-118.5 μm in length and 12.5-15.0 μm in width, with mostly 3 to 14 pseudosepta. Conidiophores (n=20) were straight to slightly curved, unbranched, and pale to light brown in color. Based on the morphological characteristics of the colonies and conidiophores and conidia, the pathogen was identified as Corynespora cassiicola (Berk and M.A. Curtis) C.T. Wei. (Berkeley & Curtis 1968; Lu et al. 2021; Wei 1950). Genomic DNA was extracted following using modified CTAB method (Gardes and Bruns 1993) and internal transcribed spacer (ITS) region was amplified with ITS1 and ITS4 primers (White et al. 1990). The ITS sequence generated of about 553 bp and deposited in GenBank (accession no. MN954556), was found more than 99% similar to previously deposited sequences of C. cassiicola (GenBank accession nos. MN339671, EU364535, FJ852574, MK139711, EU131374) as verified through BLASTn and phylogenetic tree construction. A pathogenicity test was performed for fulfilling Koch'spostulates. Conidial suspension (105 conidia/ml) of the recovered isolate was sprayed on the 5 healthy leaves of 2-month-old seedling of J. sambac. Mock inoculated plants sprayed sterile distilled water were used as a control. The seedlings were covered with plastic bags to maintain high humidity at 24 to 28°C for a week. Identical disease symptoms to those observed in nursery plants were observed on the leaves of the inoculated plants in 7 days but not mock inoculated plants and results were reconfirmed. The reoccurred fungus was isolated from the diseased spots of the inoculated leaves to complete Koch's postulates and identified microscopically. A representative sample of leaves with lesions was deposited in the LAH herbarium, Department of Botany University of the Punjab, Pakistan (LAH35691). Previously, C. cassicola has been found infecting Jasminum mesnyi in China and Jasminum sp. in Florida (Alfieri et al. 1984; Zhang et al. 2018). The best of our knowledge, this is the first report of leaf spot caused by C. cassiicola on J. sambac in Pakistan. It will establish a foundation for future studies of management strategies for this plant disease caused by C. cassiicola.

Didymosphaeriaceae is a ubiquitous fungal family that is reported to include saprobic, endophytic and pathogenic species associated with a wide variety of substrates. The family is characterized by 1-septate ascospores and trabeculate pseudoparaphyses, mainly anastomosing above the asci. In recent treatments Appendispora, Didymosphaeria, Roussoella, Phaeodothis and Verruculina were placed in the family. The aim of the present study is to delineate phylogenetic lineages within Didymosphaeriaceae and allied genera. A new species, Didymosphaeria rubi-ulmifolii, was isolated and identified based on morphological characters and phylogenetic analyses of partial 18S nrDNA and 28S nrDNA nucleotide sequence data. Didymosphaeria rubi-ulmifolii clustered with Montagnulaceae as a separate genus, while two putative strains (HKUCC 5834 and CMW 22186) of D. futilis from GenBank clustered with Cucurbitariaceae and Didymellaceae, respectively. The new species is characterized by immersed to slightly erumpent ascomata immersed under a clypeus, a peridium with compressed cells of textura intricata, long trabeculate pseudoparaphyses, anastomosing mostly above the asci and brown, 1-septate ascospores with granulate ornamentation. Phylogenetic analysis in combination with morphology and a review of literature show that Appendispora, Phaeodothis, Roussoella and Verruculina should be excluded from the family. Phaeodothis belongs in Montagnulaceae, Verruculina in Testudinaceae, while Appendispora and Roussoella belong in Roussoellaceae. The position of Didymosphaeriaceae as a distinct family, based on 1-septate ascospores and trabeculate pseudoparaphyses, mainly anastomosing above the asci is doubtful. Fresh collections of more Didymosphaeria strains are needed for epitypification and to obtain sequence data to establish if this family can be maintained.

The Amazonian fish Colossoma macropomum harbors high myxosporean diversity: A description of three novel species from a fish farm

Henneguya Thélohan, 1892is the second most species rich genus of myxozoans, with reports from freshwater and marine fish worldwide. In the Great Lakes region of North America, muskellunge Esox masquinongy is an important game fish species that serves as an apex predator in the ecosystems of many inland lakes. The myxozoan fauna of esocid fish, especially muskellunge, remains largely understudied. During fish health assessments, muskellunge were examined for parasitic infections and myxozoan pseudocysts were observed on gill clip wet mounts. When ruptured under pressure, the intralamellar pseudocysts released thousands of myxospores consistent with those of the genus Henneguya. The myxospores were 67.3-96.6 (79.1 ± 5.9) µm in total length. The spore body was 18.6-22.6 (20.9 ± 1.0) µm × 5.4-6.9 (6.3 ± 0.4) µm in valvular view and 3.5-4.0 (3.8 ± 0.3) µm wide in sutural view. The two pyriform polar capsules positioned at the anterior of the spore body were 6.4-7.7 (7.0 ± 0.4) µm × 1.8-2.1 (2.0 ± 0.1) µm and each contained a tightly coiled polar filament with 9-10 turns. Two tapering caudal processes extended from the posterior of the spore body and were 47.3-75.6 (58.3 ± 5.8) µm in length. Histologically, large intralamellar polysporic plasmodia were surrounded by plump pillar cells and a distinct layer of plasma. Mild inflammation was present peripherally, with small numbers of necrotic germinative cells and intraplasmodial phagocytes internally. Ribosomal 18S rRNA gene sequence data were obtained from three gill pseudocysts. The three ~2000-bp sequences were identical, but shared no significant similarity with any publicly available sequence data. Phylogenetic analyses demonstrated sequence data from this Henneguya fell within a well-supported clade of Henneguya spp. reported from northern pike Esox lucius in Europe. Based on the distinct morphological, histological and molecular data, this species is designated as Henneguya michiganensis n. sp. from muskellunge in Michigan, USA.

Phylogenetic analyses of the Lyophylleae (Agaricales, Basidiomycota) based on nuclear and mitochondrial rDNA sequences

During fungal surveys between 2012 and 2014 in pine-dominated forests of the western Himalayas in Pakistan, several collections of Pseudosperma (Agaricales, Inocybaceae) were made. These were documented, based on morphological and molecular data. During this work, three new species came to light, which are here formally described as Pseudospermabrunneoumbonatum, P.pinophilum and P.triacicularis. These species belong in the genus PseudospermafideMatheny et al. (2019) = Pseudosperma clade fideMatheny (2005) = Inocybe sect. Rimosaes.s.fideLarsson et al. (2009). Macro- and micro-morphological descriptions, illustrations and molecular phylogenetic reconstructions of the studied taxa are provided. The new species are differentiated from their close relatives by basidiospore size and colouration of basidiomata. Molecular phylogenetic relationships are inferred using ITS (ITS1–5.8S–ITS2), nrLSU and mtSSU sequence data. All three newly-described taxa likely share an ectomycorrhizal association with trees in the genus Pinus. In addition, five names are recombined in Inosperma, Mallocybe and Pseudosperma. These are Inospermavinaceobrunneum, Mallocybeerratum, Pseudospermaalboflavellum, Pseudospermafriabile and Pseudospermaneglectum.

Bacidia schweinitzii is a common crustose lichen that is widespread in eastern North America. It is comprised of three distinct morphotypes differing in apothecial pigmentation. Here we show that molecular data from the mtSSU region affirms the distinctiveness of these morphotypes, prompting the recognition of three species: B. schweinitzii s.str., B. ekmaniana sp. nov. and B. purpurans sp. nov. We also show that a common sorediate crustose lichen, sympatric with B. schweinitzii, represents a monophyletic lineage whose relationship with B. schweinitzii s.str. could not be resolved with certainty using analyses of ITS and mtSSU sequence data. We recognize this sorediate lineage as a distinct species, B. sorediata sp. nov. All four taxa are described, illustrated and mapped.

The lichenicolous species Sclerococcum glaucomarioides growing on the lichen Ochrolechia akagiensis is newly reported from China. This study provides an illustration and morphological description for this species, complemented by phylogenetic analyses based on ITS, LSU and mtSSU sequence data. The taxon is characterized by adpressed to sessile apothecia that are round to angular with black disc, hymenium with brownish color, 25–40 μm high, broadly cylindrical to subclavate 8-spored asci, 20–30 × 10–15 μm, with euamyloid apical cap and brown ascospores with 2–3 transverse septa.

Megalaria allenae, a blue-gray to green-blue sterile sorediate crustose lichen containing atranorin, fumarprotocetraric acid and zeorin, is described as new to science. It is corticolous on deciduous trees and endemic to swamp forests of the Atlantic Coastal Plain of southeastern North America. As sexual reproductive structures are unknown in the species, the generic placement and relationship with M. pulverea were determined using molecular phylogenetic analyses of ITS and mtSSU sequence data.

Leprocaulon beechingii is described as new to science based on collections from exposed rock outcrops in the southern Appalachian Mountains in eastern North America. Taxonomic placement in Leprocaulon, and delimitation from other members of the genus with usnic acid, is supported by molecular phylogenetic analyses of ITS and mtSSU sequence data. The species is readily recognized by its occurrence on non-calcareous rocks, normandinoides-type placodioid thallus, and the production of usnic acid and zeorin.

A review of the crustose lichen genus Phlyctis in North America is presented derived from large-scale studies of chemical, morphological, and molecular data (ITS and mtSSU). Five species are recognized based on a combination of morphological and chemical characters, namely P. agelaea, P. argena, P. boliviensis, P. speirea and the newly described P. petraea. Analyses of molecular data supported the recognition of P. boliviensis and P. petraea but recovered P. agelaea, P. argena or P. speirea in a single clade with little internal phylogenetic structure, suggesting their circumscription based on reproductive mode and thallus morphology requires further study. An account is provided that includes an identification key, discussion of each species, distribution maps and illustrations of morphology. Phlyctis petraea, provisionally named more than a decade ago, is described as new to science. Molecular phylogenetic analyses using ITS and mtSSU sequence data both confirm the placement of P. petraea in Phlyctis and demonstrate two known chemotypes (one with norstictic acid and one with norstictic and stictic acids) are not reciprocally monophyletic and should be treated as variants of a single species. Psathyrophlyctis, described as a member of the Phlyctidaceae, is discussed and treated as a member of the Lecanorales incertae sedis.

Previous morphological and histological data are supplemented with molecular and ultrastructural data for a Henneguya sp. isolated from farm-raised channel catfish Ictalurus punctatus in Mississippi, USA. Myxospores were cryptic, encapsulated within a thin layer of epithelium in the gill lamellae with spore measurements consistent with the original description of Henneguya postexilis Minchew, 1977. Myxospores were 42.7-49.1 µm in total length with spore bodies 12.1-17.2 × 3.6-4.8 × 2.9-3 µm. Polar capsules were of unequal length, with the longer capsule being 4.4-6.7 × 1.1-1.6 µm and the shorter capsule being 4.4-6.4 × 1.1-1.6 µm. Polar tubules had 6-8 turns. Caudal processes were 25.7-38.1 µm in length. Spores were encapsulated in a thin layer of epithelium in the gill lamellae. Molecular data from the most commonly used markers for myxozoan identification and phylogeny, partial 18S small subunit ribosomal gene (SSU), partial 28S large subunit ribosomal gene (LSU), and elongation factor 2 (EF2) were generated for H. postexilis. Additionally, novel data for LSU and EF2 were generated for archived myxozoan specimens from farm-raised catfish (H. mississippiensis, H. ictaluri, H. exilis, H. adiposa, H. sutherlandi, H. bulbosus, Unicauda fimbrethilae), as well as archived specimens from wild fish (H. laseeae [from Pylodictis olivaris], Hennegoides flockae [from Aphredoderus sayanus], Myxobolus cloutmani [from Cycleptus elongatus]. These include the first EF2 sequence data for the genera Hennegoides and Unicauda. Phylogenetic analyses using these data placed H. postexilis in well supported clades with other ictalurid-infecting Henneguya species. Phylogenetic signal assessments on these analyses suggest that while SSU provided the greatest phylogenetic signal, LSU yielded comparable signal, supporting previous work implying this region may be underutilised in reconstructing myxobolid phylogenies.

Two new species of the lichenized genus Lasioloma are described from Asia: Lasioloma longiramosum W. C. Wang & A. Abas (collected from Malaysia), is characterized by a distinct woolly prothallus between dispersed thallus patches, comparatively small, muriform ascospores, long filiform conidia (main branch 22–28 μm in length, the other three branches 65–80 μm) and a foliicolous habitat; L. verrucosum W. C. Wang & X. L. Wei (collected from China), is characterized by a warted thallus, filiform conidia (main branch 22–32 μm in length, the other three branches 50–65 μm) and a corticolous habitat. The placement of both new species was confirmed by a molecular phylogenetic approach based on combined ITS, mtSSU and mtLSU sequences, and both are compared in detail to other similar species of the genus. Our study also revealed that the length of the conidial branches, which has not been explored in previous studies, should be regarded as an important feature for species delimitation in Lasioloma.