Characterization of a Cu2+, SDS, alcohol and glucose tolerant GH1 ß-glucosidase from Bacillus sp. CGMCC 1.16541.

A ß-glucosidase gene (bsbgl1a) from Bacillus sp. CGMCC 1.16541 was expressed in Escherichia coli BL21 and subsequently characterized. The amino acid sequence shared 83.64% identity with ß-glucosidase (WP_066390903.1) from Fictibacillus phosphorivorans. The recombinant ß-glucosidase (BsBgl1A) had a molecular weight of 52.2 kDa and could hydrolyze cellobiose, cellotriose, cellotetrose, p-nitrophenyl-ß-D-glucopyranoside (pNPG), and p-nitrophenyl-ß-D-xylopyranoside (pNPX). Optimal activity for BsBgl1A was recorded at 45 °C with a pH between 5.6 and 7.6, and 100% of its activity was maintained after a 24 h incubation between pH 4 and 9. Kinetic characterization revealed an enzymatic turnover (Kcat) of 616 ± 2 s-1 (with cellobiose) and 3.5 ± 0.1 s-1 (with p-nitrophenyl-ß-D-glucopyranoside). Interestingly, the recombinant enzyme showed cupric ion (Cu2+), sodium dodecyl sulfate (SDS) and alcohol tolerance at 10 mM for Cu2+ and 10% for both SDS and alcohol. Additionally, BsBgl1A had high tolerance for glucose (Ki = 2095 mM), which is an extremely desirable feature for industrial applications. Following the addition of BsBgl1A (0.05 mg/ml) to a commercial cellulase reaction system, glucose yields from sugarcane bagasse increased 100% after 1 day at 45 °C. This work identifies a Cu2+, SDS, alcohol, and glucose tolerant GH1 ß-glucosidase with potential applications in the hydrolysis of cellulose for the bioenergy industry.

New Species and Records of Pleurotheciaceae from Karst Landscapes in Yunnan Province, China.

Pleurotheciaceae is a genera-rich and highly diverse family of fungi with a worldwide distribution in aquatic and terrestrial habitats. During the investigation of lignicolous freshwater fungi from karst landscapes in Yunnan Province, China, 15 fresh strains were obtained from submerged decaying wood. Based on the morphology and phylogenetic analysis of a combined LSU, ITS, SSU, and rpb2 sequence dataset, Obliquifusoideum triseptatum, Phaeoisaria obovata, Pleurotheciella brachyspora, Pl. longidenticulata, and Pl. obliqua were introduced as new species, P. synnematica and Rhexoacrodictys melanospora were reported as new habitat records, and P. sedimenticola and Pl. hyalospora were reported as new collections. In addition, based on morphological comparisons and phylogenetic analysis, we accepted Obliquifusoideum into in the family Pleurotheciaceae (Pleurotheciales, Savoryellomycetidae). Freshwater habitats are the primary habitats of Pleurotheciaceae species, and Yunnan Province has the highest concentration and species diversity of Pleurotheciaceae in China.

Morphological and Phylogenetic Analyses Reveal Three Novel Species of Sanguinoderma (Ganodermataceae, Basidiomycota) from Yunnan Province, China.

Sanguinoderma (Ganodermataceae) is recognized as a valuable medicinal resource in Taiwan, China. Additionally, it serves as a traditional folk medicine for treating neurotic epilepsy in Malaysia. This study involved the collection of six specimens of Sanguinoderma from Yunnan Province, China. Employing multigene phylogenetic analysis of DNA sequences, including internal transcribed spacer (ITS), nuclear large subunit (LSU), RNA polymerase II second largest subunit (rpb2), translation elongation factor 1-alpha (tef1-α), mitochondrial small subunit (mtSSU), nuclear small subunit (nSSU) and morphological examinations, three new species, viz. Sanguinoderma concentricum, S. dehongense and S. ovisporum, are introduced. Sanguinoderma concentricum is characterized by a central stipe basidiomata, an orbicular to suborbicular pileus, a grayish-yellow surface with alternating concentric zones and wavy margin-like petals and regular pileipellis cells (4-8 × 17-28 µm). Sanguinoderma dehongense is characterized by a long stipe and flabelliform basidiomata, a dark-grayish yellow-to-dark-yellow pileus surface, irregular pileipellis cells and wavy margin and ellipsoid basidia (8-11 × 9-13 µm). Sanguinoderma ovisporum is characterized by a reniform basidiomata, a heterogeneous context and ovoid basidiospores (7.5-8.6 × 5.5-7.2 µm). A detailed description and illustrations of these new species are provided, as well as a morphological comparison with similar taxa.

Three New Species of Tuber Discovered in Alpine Fir Forests in Yunnan, China.

Three new species of Tuber, T. albicavum, T. laojunshanense, and T. umbilicicavatum belonging to the Puberulum phylogroup, are described based on specimens collected in alpine Abies forests at 3600-4000 m, Northwest Yunnan, China. T. albicavum is distinguished by its ascomata with a single chamber of 0.5-1.8 cm diameter, with an apical opening of 0.2-0.6 cm in diameter, and light golden-brown alveolate reticulate ascospores up to 30 µm in length; T. laojunshanense is characterized by having ascomata with a slightly tomentose surface, sometimes with a white navel, a relatively thick peridium, up to 280 µm, and yellow-brown spores with alveolate reticulate ornamentation, up to 34 µm in length; T. umbilicicavatum is characterized by smooth ascomata with a distinct white navel, a relatively thin peridium, up to 110 µm, and golden or golden-brown alveolate reticulate ascospores, up to 40 µm in length. The molecular analysis of the internal transcribed spacer region also supports that these three new species differ from previously described Tuber species.

Diversity of Distoseptispora (Distoseptisporaceae) taxa on submerged decaying wood from the Red River in Yunnan, China.

The Red River Basin is located in the Indo-Burma biodiversity hotspot and is rich in lignicolous freshwater fungi, but no systematic research has been conducted. A systematic study on the species diversity of lignicolous freshwater fungi in the basin is ongoing. Seven distoseptispora-like specimens were collected from the Red River Basin in Yunnan. Phylogenetic analysis of ITS, LSU, tef1-α, and rpb2 genes and combined morphological data indicate that there are six distinct species of Distoseptispora, including two new species and four known species. Two new species were named D.suae and D.xinpingensis, and the four known species were D.bambusae, D.euseptata, D.obpyriformis and D.pachyconidia. This study provides detailed descriptions and illustrations of these six species and an updated phylogenetic backbone tree of Distoseptispora.

New species and new records of Laccaria (Agaricales, Basidiomycota) from Northern Thailand.

Two new species Laccariapseudoalba and L.subroseoalbescens are described and illustrated, based on morphological characteristics and molecular phylogenetic analysis. Two new records, Laccariaumbilicata and L.yunnanensis from Thailand, are also reported. Laccariasubroseoalbescens is characterized by small basidiomata, stipe equal with an enlarged base, and nearly subclavate, pale pink to light orange. Laccariapseudoalba is characterized by pale orange to orange white pileus, has umbo when young on the pileus, and fistulose stipe of the pale to pastel red color. Phylogenetic analysis based on sequence data from rDNA internal transcribed spacer ITS1-5.8S-ITS2 rDNA (ITS), nuc 28S rDNA (28S), RNA polymerase II subunit 2 (rpb2), and translation elongation factor 1-α (tef1-α) are provided as further evidence. Molecular analysis confirms the phylogenetic positions of the two new species and two new records. The differences in characteristics of these two new species and closely related species are discussed herein.

Two new species of Ganoderma (Ganodermataceae, Basidiomycota) from Southwest China.

Ganoderma is a large and diverse genus containing fungi that cause white rot to infect a number of plant families. This study describes G.phyllanthicola and G.suae as new species from Southwest China, based on morphological and molecular evidence. Ganodermaphyllanthicola is characterized by dark brown to purplish black pileus surface with dense concentric furrows, pale yellow margin, irregular pileipellis cells, small pores (5-7 per mm) and ellipsoid to sub-globose basidiospores (8.5-10.0 × 6.0-7.5 µm). Ganodermasuae is characterized by reddish brown to oxblood red pileus surface and lead gray to greyish-white pore surface, heterogeneous context, wavy margin and almond-shaped to narrow ellipsoid basidiospores (8.0-10.5 × 5.0-7.0 µm). The phylogeny of Ganoderma is reconstructed with multi-gene sequences: the internal transcribed spacer region (ITS), the large subunit (nrLSU), translation elongation factor 1-α gene (TEF-1α) and the second subunit of RNA polymerase II (RPB2). The results show that G.suae and G.phyllanthicola formed two distinct line-ages within Ganoderma. Descriptions, illustrations and phylogenetic analyses results of the two new species are presented.

Morphology and multi-gene phylogeny reveal a novel Torula (Pleosporales, Torulaceae) species from the plateau lakes in Yunnan, China.

Background: During an investigation into lignicolous freshwater fungi from the plateau lakes in Yunnan Province, China, two fresh collections of Torula taxa were collected and examined morpholgically. New information: Torulaluguhuensis is characterised by: conidiophores which are semi-macronematous mononematous, erect, septate, smooth, slightly flexuous and pale brown; conidiogenous cells which are holoblastic, mono- to polyblastic, integrated, terminal, terminal or intercalary in conidial chains, doliiform and pale brown; conidia which are branched chains, acrogenous, straight or slightly curved, dark brown to blackish, pale brown or subhyaline at apex, 1-3 septate, strongly constricted at the septa, verruculose or finely echinulate and rounded at both ends. A new species was introduced, based on morphological and phylogenetic analysis of combined ITS, LSU, RPB2 and TEF sequence data. Detailed descriptions and illustrations are provided, with an updated phylogenetic tree depicting intergeneric relationships within the Torulaceae.

Additions to the Genus Helicosporium (Tubeufiaceae, Tubeufiales) from China with an Identification Key for Helicosporium Taxa.

Helicosporous hyphomycetes is a group of filamentous fungi that shows promising application prospects in metabolizing bioactive natural compounds. During a study of helicosporous fungi in China, six new helicosporous taxa were collected and isolated from decaying wood in Guangxi Zhuang Autonomous Region, China. Morphological comparisons with multi-gene phylogenetic analyses revealed that these six taxa belong to Helicosporium (Tubeufiaceae, Tubeufiales), and they were recognized as three novel species and were named Helicosporium liuzhouense, H. multidentatum, and H. nanningense. Detailed descriptions and illustrations of the newly discovered taxa and comparisons with similar fungi are provided. In addition, a list and a key to accepted Helicosporium species are provided.

Two novel species and three new records of Torulaceae from Yunnan Province, China.

While investigating the diversity of lignicolous fungi in Yunnan Province, China, six fresh collections of Torulaceae were collected and identified based on morphological examination and phylogenetic analyses of combined LSU, ITS, SSU, tef1-α, and rpb2 sequence data. Two new species, viz. Neopodoconisyunnanensis and Torulasuae, and three new records, viz. T.canangae (new freshwater habitat record), T.masonii (new host record), and T.sundara (new freshwater habitat record) are reported. Detailed descriptions, illustrations, and a phylogenetic tree to show the placement of these species are provided.

Morphological and phylogenetic analyses reveal three new species and one new record of Tubeufia (Tubeufiales, Tubeufiaceae) from southern China.

During an investigation of helicosporous fungi in China, a total of seven helicosporous hyphomycetes were obtained from decaying wood in the southern region of the country. Based on phylogenetic analyses using a combined LSU, ITS, tef1α, and rpb2 sequence matrix, in conjunction with morphological comparisons, these taxa were classified within Tubeufia (Tubeufiaceae, Tubeufiales) and were recognized as three new species, viz. Tubeufiaguttulata, T.hainanensis, and T.muriformis, as well as one new distribution record, viz. T.cocois. Evidence for these new taxa and the new record, descriptions, illustrations, notes, and phylogenetic evidence are provided for the newly collected helicosporous species.

Lignicolous Freshwater Fungi from Plateau Lakes in China (I): Morphological and Phylogenetic Analyses Reveal Eight Species of Lentitheciaceae, Including New Genus, New Species and New Records.

During the investigation of lignicolous freshwater fungi in plateau lakes in Yunnan Province, China, eight Lentitheciaceae species were collected from five lakes viz. Luguhu, Qiluhu, Xingyunhu, Cibihu, and Xihu lake. Based on morphological characters and phylogenetic analysis of combined ITS, LSU, SSU, and tef 1-α sequence data, a new genus Paralentithecium, two new species (Paralentithecium suae, and Setoseptoria suae), three new records (Halobyssothecium phragmitis, H. unicellulare, and Lentithecium yunnanensis) and three known species viz. Halobyssothecium aquifusiforme, Lentithecium pseudoclioninum, and Setoseptoria bambusae are reported.

New species of Hydnotrya (Ascomycota, Pezizomycetes) from southwestern China with notes on morphological characteristics of 17 species of Hydnotrya.

More specimens of Hydnotrya have been collected from southwestern China in recent years. Morphological and molecular analyses showed that they belonged to three species of Hydnotrya, of which two are new to science, H.oblongispora and H.zayuensis. The third one was H.laojunshanensis, previously reported in 2013. The new species are described, and their relationship to other species of Hydnotrya is discussed. H.laojunshanensis is re-described in more detail. The main morphological characters of 17 species of Hydnotrya are compared and a key to them is provided as well.

Geastrumsuae sp. nov. (Geastraceae, Basidiomycota) a new species from Yunnan Province, China.

Background: Geastrum is the largest genus of Geastraceae and is widely distributed all over the world. Four specimens which belong to Geastrum were collected during our scientific expedition to Cangshan Mountain, Yunnan, China. Based on morphological characteristics and phylogenetic analysis, a new species was introduced. New information: Geastrumsuae is characterised by its large basidiomata (height 35-70 mm, diameter 18-37 mm) with long stipe (height 10-45 mm), smooth pink exoperidium and sessile globose endoperidial body. Phylogenetic analysis has been carried out, based on the internal transcribed spacer (ITS) and large subunit ribosomal ribonucleic acid (nrLSU) sequence data. The illustration and description for the new taxa are provided.

Four novel Pleurocordyceps (Polycephalomycetaceae) species from China.

Entomopathogenic fungi comprise an ecologically important group of specialized pathogens infecting other fungi, invertebrates, and plants. These fungi are species-rich with high diversity and broad distribution worldwide. The majority of entomopathogenic fungi belong to clavicipitoids, which consist of the hypocrealean families, Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae, and Polycephalomycetaceae. The latter is a newly established entomopathogenic family that recently separated from the family Ophiocordycipitaceae to accommodate the genera, Perennicordyceps, Pleurocordyceps, and Polycephalomyces. In recent years, Polycephalomycetaceae has been enriched with parasitic and hyperparasitic fungi. With 16 species spread across China, Ecuador, Japan, and Thailand, Pleurocordyceps is the most speciose genus in the family. In this study, we expand the number of taxa in the genus by introducing four new Pleurocordyceps species from China, namely, P. clavisynnema, P. multisynnema, P. neoagarica, and P. sanduensis. We provide detailed descriptions and illustrations and infer genus-level phylogenies based on a combined 6-loci gene sequence dataset comprising the internal transcribed spacer gene region (ITS), small subunit ribosomal RNA gene region (SSU), large subunit rRNA gene region (LSU), translation elongation factor 1-alpha gene region (TEF-1α), RNA polymerase II largest subunit gene region (RPB1), and RNA polymerase II second largest subunit (RPB2). This study contributes to knowledge with regard to the diversity of Pleurocordyceps specifically and entomopathogenic Hypocreales more broadly.

Lignicolous freshwater fungi in Yunnan Province, China: an overview.

Yunnan Province is one of the rich biodiversity hotspots with abundant resources of lignicolous freshwater fungi. A total of 281 species of lignicolous freshwater fungi from 1986 to the present in Yunnan Province. They are mostly distributed in the classes Dothideomycetes and Sordariomycetes, a few species in the Eurotiomycetes and Leotiomycetes, and rarely reported in Orbiliomycetes and Pezizomycetes. Lignicolous freshwater fungi can decompose lignocellulose substrates and release energy and nutrients, and thus playing an important role in freshwater environment. This study briefly reviewed the biodiversity and taxonomic status of lignicolous freshwater fungi in Yunnan, the ecological functions of lignicolous freshwater fungi, factors affecting community distribution, application status, and research difficulties.

Lignicolous freshwater ascomycetes from Thailand: Introducing Dematipyriformamuriformis sp. nov., one new combination and two new records in Pleurotheciaceae.

During the study of lignicolous freshwater fungi from Thailand, three pleurotheciaceous species were collected from freshwater habitats in Thailand. Two were identified as Pleurotheciumaquaticum and Rhexoacrodictysfimicola, and the third is a new species Dematipyriformamuriformis sp. nov.. Rhexoacrodictys is accepted in Pleurotheciaceae based on phylogenetic analysis. Rhexoacrodictysnigrospora is transferred to Dematipyriforma based on phylogenetic analysis and morphological characters. Pleurotheciumaquaticum and Rhexoacrodictysfimicola are reported from Thailand for the first time.

Novel Species and Records of Dictyosporiaceae from Freshwater Habitats in China and Thailand.

China and Thailand are rich in fungal diversity with abundant freshwater resources that are favorable for numerous fungal encounters. Resulting from this, the majority of the Dictyosporiaceae species reported were from these two countries. During the investigation on the diversity of lignicolous freshwater fungi in the Greater Mekong Subregion, eleven collections of cheirosporous species on submerged wood were collected from lentic and lotic habitats in China and Thailand. Phylogenetic analysis that combined nuclear small-subunit ribosomal RNA (SSU), internal transcribed spacer region (ITS), nuclear large subunit ribosomal RNA (LSU) and translation elongation factor 1α (tef 1-α) loci revealed six new species: Dictyocheirospora chiangmaiensis, D. multiappendiculata, D. suae, Digitodesmium aquaticum, Vikalpa grandispora and V. sphaerica. In addition, four known species were also identified and reported based on morphological and phylogenetic evidence. The detailed descriptions and illustrations of these taxa are provided with an updated phylogenetic tree of Dictyosporiaceae.

Neomonodictysaquatica sp. nov. (Pleurotheciaceae) from a plateau lake in Yunnan Province, China.

BACKGROUND: In this study, a new species Neomonodictysaquatica was collected from submerged decaying wood in Erhai Lake, Yunnan Province, China. NEW INFORMATION: Neomonodictysaquatica is characterised by acrogenous, solitary, oval, dictyospores (most are transverse septum, occasionally vertical septum, in immaturity the septum is clear, but when mature, the conidia becomes darker so the septum is not clear), smooth-walled conidia. The immature conidia are usually hyaline to olivaceous and mature conidia are usually darkened to black, sometimes with one pale basal cell. Phylogenetic analyses of combined ITS and LSU sequence data showed that the new collection is distinct from other Neomonodictys species. Description and illustration are provided as well.

Endophytic Pestalotiopsis species associated with Rhododendron in Cangshan Mountain, Yunnan Province, China.

Rhododendron is an essential ornamental plant that is abundant in Yunnan province. In Cangshan Mountain, Yunnan, China, 61 species of Rhododendron have been reported. Endophytic fungi are internal plant tissue inhabitants that do not harm the host. It has emerged as an exciting research topic as they have the potential to provide numerous secondary metabolites. This study is focused on taxonomic novelties and new host records of endophytic fungi associated with Rhododendron plants collected from Cangshan Mountain in Yunnan Province, China. Pestalotiopsis fungi are associated with a vast array of plant species worldwide. In this study, fresh leaves of Rhododendron cyanocarpum, Rhododendron decorum, and Rhododendron delavayi were collected from Cangshan Mountain, Yunnan Province, China. Endophytic Pestalotiopsis fungi associated with Rhododendron were characterized based on phylogenetic analyses of combined ITS, TEF1-α, and TUB genes along with morphological characteristics. Six new species (Pestalotiopsis appendiculata, Pestalotiopsis cangshanensis, Pestalotiopsis daliensis, Pestalotiopsis fusoidea, Pestalotiopsis rosarioides, and Pestalotiopsis suae) and a new host record (Pestalotiopsis trachicarpicola) are described. Detailed descriptions and color photo plates of these species are provided. It is the first time that the endophytic fungi of Rhododendron plants in Cangshan Mountain have been studied.