Morphology, Phylogeny and Culture Characteristics of Ganoderma gibbosum Collected from Kunming, Yunnan Province, China

Ganoderma is a genus of medicinally and economically important mushrooms in the family Ganodermataceae.Ganoderma species are popular medicinal mushrooms and their health benefits are well-documented.Ganoderma is a cosmopolitan genus that is widely distributed in both tropical and temperate regions.This genus is characterized by its unique laccate or non-laccate species with double-walled basidiospores.Here,we report on eight collections of G.gibbosum collected during surveys in Kunming,Yunnan Province,China.The specimens are described and illustrated based on macro-and micro-morphological characteristics.Total DNA of the eight G.gibbosum strains were extracted using the Biospin Fungal Extraction Kit following manufacturer protocol.Amplification of the Internal Transcribed Spacer(nrITS)region was carried out using ITS5/ITS4 primers and LROR/LR5 for the nuclear ribosomal large subunit 28S rDNA gene(LSU).Phylogenetic analysis with closely related species to G.gibbosum showed that all eight collections grouped with G.gibbosum with 100%bootstrap support.Phylogenetic similarity and morphological variations within the eight collections of G.gibbosum are discussed.

Multi-gene phylogeny of Jattaea bruguierae,a novel asexual morph from Bruguiera cylindrica

During our survey on marine-based ascomycetes of southern Thailand,fallen mangrove twigs were collected from the intertidal zones.Those specimens yielded a novel asexual morph of Jattaea(Calosphaeriaceae,Calosphaeriales),Jattaea bruguierae,which is confirmed as a new species by morphological characteristics such as nature and measurements of conidia and conidiophores,as well as a multigene analysis based on combined LSU,SSU,ITS andβ-tubulin sequence data.Jattaea species are abundantly found from wood in terrestrial environments,while the asexual morphs are mostly reported from axenic cultures.Jattaea bruguierae is the first documentation of an asexual morph species from marine-habitats.

First successful domestication of a white strain of Auricularia cornea from Thailand

Intraspecies colour variations in cultivated edible mushrooms present novel and potentially valuable alternatives to the research and cultivation industries.In this study,we collected,identified,and domesticated a white strain of Auricularia cornea from Thailand.The brown strain of A.cornea is one of the top two species of Auricularia cultivated and traded in Asia.Since both white and brown phenotypes of A.cornea belong to a single species,we established their similarities or differences.Both morphological characteristics and phylogenetic analysis of ITS rDNA sequence data were used to confirm the taxonomic placement of the white A.cornea strain in the same clade with the brown A.cornea.Nutritional analysis showed that fat,fiber,protein,and total soluble sugar contents of the white A.cornea were significantly higher than the commercially used brown strain.The melanin content of the white strain of A.cornea(less than 1.5 mg/100g)was not significantly different from that of the brown strain.This discovery may create new opportunities for the mushroom growing industry and for smallholder farmers in Asia.

Sexual Morph of Furcasterigmium furcatum (Plectosphaerellaceae) from Magnolia liliifera Collected in Northern Thailand

We isolated an interesting fungus from dead leaves of Magnolia liliifera collected from Chiang Mai,Thailand.The novel strain is related to Plectosphaer-ellaceae based on the morphology of its asexual morph and the analysis of sequence data.Phylogenetic analyses using a combined gene analysis of LSU and ITS sequence data showed that this strain is clustered in the same clade with Furcasterigmium furcatum with high statistical support.The new strains produced the asexual morph in culture which is morphologically similar to F.furcatum.Thus,we identified this strain as the sexual morph of F.furcatum.This is the first record of sexual morph for the monotypic genus Furcasterigmium and the first record of this genus on Magnolia.

Neohelicosporium fusisporum sp.nov.(Tubeufiaceae)and a first record of a sexual morph within Neohelicosporium

Both sexual and asexual morphs(holomorph)are known in several genera of Tubeufiaceae whereas in others,mainly the asexual or rarely the sexual morph is known.The genus Neohelicosporium is known only by its asexual morph,we have recently collected the sexual morph of the genus,which is characterized by fleshy,superficial ascomata,bitunicate asci,and hyaline to pale brown,fusiform ascospores.Analyses of combined ITS,LSU and tef1 sequence data placed the taxon in Neohelicosporium.In this paper,we introduce the new taxon N.fusisporum based on both morphological characteristics and phylogenetic data.

A review of Entoloma sensu lato(Basidiomycota,Entolomataceae)from Yunnan Province,China

This review succinctly discusses data on morphology,systematics,ecology and biochemical properties of all the Entoloma species from Yunnan Province,China.We also propose directions for future research on Entoloma.Altogether,8 species of Entoloma have been reported in the present study.Entoloma sinuatum and E.mengsongense showed the most contrasting variation in colour and size of the basidiomes while E.mengsongense and E.yunnanense were found to display wide variations in spore morphology.Most species are saprotrophic except E.caeruleoflavum and E.sinuatum which form mycorrhizal associations.Knowledge on biochemical and other chemical attributes of the genus are minimal although such details would help for better utilization of the genus.

Morphological and molecular identification of Panus conchatus(Polyporaceae,Polyporales)from Yunnan Province,China

Panus conchatus is a species of lentinoid fungi in the family Polyporaceae.This species is characterized by its concave,smooth,deeply decurrent gills,with distinctive purple grey to greyish magenta basidiocarps.This fungus is widely distributed in both tropical and temperate regions.Here,we report a specimen of P.conchatus collected from a temperate region in Yunnan Province,China.The specimen is described and illustrated based on macro-and micro-morphological characteristics.Phylogenetic analyses were done based on the sequence data of ITS and LSU,and the placement of the taxon was confirmed.This is the first time P.conchatus has been reported with molecular phylogenetic data from China.Full description,illustrations,color photographs,and a phylogenetic tree to show the placement of P.conchatus are provided.

Saprobic Dothideomycetes in Thailand:Muritestudina gen.et sp.nov.(Testudinaceae)a new terrestrial pleosporalean ascomycete,with hyaline and muriform ascospores

The family Testudinaceae and its intergeneric classification are poorly understood.This is due to overlap of morphological characteristics in genera and lack of DNA sequence data to infer phylogenetic relationships.The main objective of the present paper is to establish a novel genus,Muritestudina,based on distinct morphological characteristics and analyses of combined LSU,SSU,ITS,rpb2 and tef1 sequence data.We also fill the gap of our current knowledge on the phylogenetic position of Testudinaceae.Based on the morphological characteristics of species representing existing genera of Testudinaceae,we herein introduce a new genus,Muritestudina with M.chiangraiensis as the type species.The new genus is characterized by globose to subglobose,ostiolate ascomata;a peridium of brown to dark-brown cells of textura angularis;septate and cellular pseudoparaphyses;cylindric-clavate asci with a distinct pedicel;and hyaline,ellipsoidal and muriform ascospores.The new genus differs from the other genera in Testudinaceae in having hyaline,muriform ascospores.Combined analyses of ribosomal and protein coding gene sequence data confirmed that our new taxon belongs in Testudinaceae with a close relationship with Neotestudina rosatii.

Saprobic Dothideomycetes in Thailand: Vaginatispora appendiculata sp. nov. (Lophiostomataceae) introduced based on morphological and molecular data

In order to establish the evolutionary relationships and resolve the polyphyletic nature of Dothideomycetes,we are studying their natural classification based on both morphology and multigene phylogeny.In this paper we introduce Vaginatispora appendiculata,a novel species on dead twigs from southern Thailand.Morphological character differences and analyses of combined LSU,TEF,SSU and ITS sequence datasets support the validity of the new species and its placement in Vaginatispora(Lophiostomataceae).

Bionectria pseudochroleuca,a new host record on Prunus sp.in northern Thailand

This study presents the first report of Bionectria pseudochroleuca(Bionectriaceae)on Prunus sp.(Rosaceae)from northern Thailand,based on both morphological characteristics and multilocus phylogenetic analyses of internal transcribe spacer(ITS)and Beta-tubulin(TUB2).

Polyurethane-degrading fungi from soils contaminated with rocket propellant and their ability to decompose alkyne terminated polybutadiene with urethane

A large amount of propellant materials are produced every year,and storage and disposal of these propellant materials seriously contributes to environmental pollution.Alkyne terminated polybutadiene with urethane segments(PUPB)is the macromolecule backbone of these propellant materials,and degradation of PUPB is central to the eco-friendly treatment of propellant materials.In this study,we isolated a polyurethane(PU)-and PUPB-degrading fungus from soils contaminated with rocket propellant,and the fungus H14 was identified as Fusarium solani(Mart.)Sacc.based on macro-and micro-morphology as well as phylogenetic analyses.The ability of F.solani H14 to degrade PU film and PUPB patches was evaluated via mass loss,scanning electron microscopy(SEM)and enzyme production ability.Mass loss analyses revealed a 25.8%reduction in mass of PU and 1.3%reduction in mass of PUPB after F.solani H14 was incubated with PU and PUPB for 90 days,respectively.We found that F.solani H14 mycelia significantly colonized both PU and PUPB.SEM images showed that the surface of PU films and PUPB patches formed holes,underwent folding and experienced damage as well as irregular fissuring from the erosion of fungal hypha.Moreover,two possible degradative enzymes,lipase and esterase,were produced by F.solani.Our study opens a new avenue of research for eco-friendly treatments of explosive materials and propellants.This paper represents the first article on the degradation of PUPB patches.

Camarosporium uniseriatum nom.nov.,from Celtis occidentalis in European Russia

Celtis occidentalis(American hackberry)is a deciduous tree widely distributed in northern America and introduced in many regions of Europe.In this study we collected Cucurbitaria celtidis from dead or dying twigs and branches of C.occidentalis(Cannabaceae)in the Rostov region(Southern European Russia),where this tree is a common ergasiophyte in artificial forests.The placement of this species in Camarosporium sensu stricto in Pleosporinae,Pleosporales is shown in a multi-locus tree based on combined LSU,SSU and ITS sequence data.Camarosporium uniseriatum nom.nov.is introduced based on morphological and phylogenetic analyses.

Ramularia coleosporii(Mycosphaerella)on Plumeria rust in Thailand

A hyperparasitic fungus was found on uredinia of Coleosporium plumeria on leaves of Plumeria rubra in Thailand.The hyperparasite was identified as Ramularia coleosporii following an examination of its morphological characters and a phylogenetic analysis by using ITS sequence data.This is the first record of R.coleosporii on C.plumeriae in Thailand.Ramularia coleosporii has the potential for biocontrol management strategies of the rust.

Additions to wild seed and fruit fungi 2:Parascedosporium putredinis:a new Thailand record from Delonix regia seed pods

We are studying seed pod fungi associated with Delonix regia in Thailand and in this paper we provide an illustrated account of Parascedosporium putredinis,which is a new record for Thailand.Phylogenetic analysis of combined ITS and LSU sequence data and morphological similarities provide evidence for its identification.We illustrate the synnematal stage and lateral,solitary conidiophores of P.putredinis obtained in vitro on MEA.We also provide ITS,LSU,SSU,and tef sequence data for P.putredinis,which are deposited in GenBank.

Linking ectomycorrhizal mushroom species richness and composition with dominant trees in a tropical seasonal rainforest

Vegetation,elevation gradient and soil temperature are considered as major drivers of ECM fungi species richness.ECM sporocarps were collected during rainy seasons for two years to study the link between the distribution of ECM mushrooms with Castonopsis echinocarpa,Parashorea chinensis,and Pittosporopsis kerrii with varying elevations and soil temperatures,in a tropical rain forest Xishuangbanna,Yunnan,China.For each tree species,60 trees of approximately the same size were selected,where half of them were growing at higher elevation levels and the rest at lower levels.The highest total counts of ECM fungi,as well as the highest species richness were produced by P.chinensis followed by C.echinocarpa and P.kerrii.Highest species richness was shown in September by P.chinensis,while P.kerrii trees had the lowest count of mushrooms across rainy seasons.Species of Boletales were recorded with highest species richness followed by species of order Agaricales around both C.echinocarpa and P.chinensis.ECM fungi count declined with increased elevation.Furthermore,fungi species richness increased positively with increased soil temperature in a tropical seasonal rainforest.

Morphology and multigene phylogeny revealed Peroneutypa aquilariae sp.nov.(Diatrypaceae,Xylariales)from Aquilaria sinensis in Yunnan Province,China

Dead twigs of Aquilaria sinensis(Thymelaeaceae)with fungal fruiting bodies were collected from Xishuangbanna,Yunnan Province,China.After initial morphological observations,an interesting fungus whose morphologically resembled Peroneutypa was isolated.Molecular phylogeny of combined ITS and tub2 showed our fungal collection is phylogenetically closely related to P.mackenziei.However,in morphology,our fungal collection is distinct from P.mackenziei in having an ostiolar canal without periphyses and the absence of paraphyses.Based on unique morphological characteristics and multigene phylogenetic analyses results,our fungal isolate is described in this paper as Peroneutypa aquilariae sp.nov.In addition,this is the first report of the genus Peroneutypa from the host A.sinensis.Full description,illustrations,and a phylogenetic tree to show the placement of the new species are provided.A synoptic table of morphological characteristics in Peroneutypa reported worldwide is also provided.

Diaporthe collariana sp.nov.,with prominent collarettes associated with Magnolia champaca fruits in Thailand

We are studying seed and fruit inhabiting fungi in Thailand and this paper introduces a new species,Diaporthe collariana,from Magnolia champaca fruits,collected in Chiang Rai Province.Molecular analysis of a combined ITS,TEF1,TUB and CAL sequence DNA and morphological data provide evidence to justify the new species.Diaporthe collariana is characterized by producing alpha and beta conidia,and conidiogenous cells with prominent,flared collarettes.The new species is compared with closely related species in the genus.

Taxonomic circumscription and phylogenetics of novel didymellaceous taxa with brown muriform spores

Sexual morph of didymellaceous taxa are characterized by their ascomata with relatively thin peridium,cylindric-clavate to clavate,short-pedicellate or apedicellate asci,hyaline to brown,1-septate to muriform ascospores.Its asexual morphs are coelomycetous and comprising pycnidial or acervulus conidiomata,phialidic,hyaline conidiogenous cells and hyaline or pale brown,septate or aseptate conidia.The majority of these cosmopolitan species are plant associated fungi which can be pathogens on a wide range of hosts and some species are of particular relevance for quarantine measures.Recent studies have significantly improved the taxonomy and systematics of didymellaceous taxa based on molecular phylogenetics.In contrast to the accurate and detailed studies on the asexual morphs which are common obligate pathogens,information on their usually saprobic sexual morphs is still limited.Among these phenotypically diverse species,spore characteristics are quite unique as most have hyaline spores with 0-1 septum,while only Neomicrosphaeropsis and Didymellocamarosporium are reported as producing pigmented,muriform spores.These dematiaceous muriform spores are characteristic of a considerable number of species that may be quite divergent in other characters.During taxonomic investigations on the diversity of didymellaceous taxa,we have isolated species from Alhagi pseudalhagi,Coronilla emerus,Cytisus sp.,Elaeagnus angustifolia and Spartium junceum in Italy,Russia and Uzbekistan.A comprehensive phylogeny,based on four loci(ITS,LSU,rpb2 and tub2)is used to infer species relationships.Comprehensive morphological descriptions and in-depth phylogenetic investigations of five new species viz.Ascochyta coronillae-emeri,Microsphaeropsis spartii-juncei,Neomicrosphaeropsis alhagi-pseudalhagi,N.cytisicola and N.elaeagni are presented.

New host and geographic records of five pleosporalean hyphomycetes associated with Musa spp.(Banana)

A study was undertaken to collect and identify saprobic fungi associated with Musa spp.(banana)from Taiwan(China),and Thailand.Samples were collected during the dry season and their morpho-molecular relationships were investigated.Five brown pleosporalean hyphomycetous taxa in Periconiaceae and Torulaceae viz.Periconia cortaderiae,P.delonicis,Torula chromolaenae,T.fici,and T.masonii were identified for the first time from Musa spp.(Musaceae).Phylogenetic analyses of a combined SSU,LSU,ITS,RPB2 and TEF DNA sequence dataset further justified the taxonomic placements of these five taxa in the above mentioned families.Periconia delonicis is reported for the first time on a monocotyledonous host and T.masonii is the first geographical record from Taiwan(China).

Direct comparison of culture-dependent and culture-independent molecular approaches reveal the diversity of fungal endophytic communities in stems of grapevine(Vitis vinifera)

Grapevines(Vitis vinifera)are colonized by ubiquitous microorganisms known as endophytes,which may have advantageous or neutral effects without causing disease symptoms.Certain endophytes are uncultivable,so culture-independent approaches such as next generation sequencing(NGS)can help for a better understanding of their ecology and distribution.To date,there are no studies which directly link NGS results with taxa derived from a culturing approach,integrating morphological and multi-gene phylogenetic analysis of endophytes.In this study,a culture-dependent and high-resolution culture-independent approach(next generation sequencing)were used to identify endophytes in grapevine stems.In the culture-dependent approach,a total of 94 isolates were recovered from 84 of 144 healthy grapevine stem fragments(colonization rate=58.3%).The study is unique as we used subsets of combined multi-gene regions to identify the endophytes to species level.Based on each multi-gene phylogenetic analysis,28 species belong to 19 genera(Acremonium,Alternaria,Arthrinium,Ascorhizoctonia,Aspergillus,Aureobasidium,Bipolaris,Botryosphaeria,Botrytis,Chaetomium,Cladosporium,Curvularia,Hypoxylon,Lasiodiplodia,Mycosphaerella,Nigrospora,Penicillium,Phoma,Scopulariopsis)were identified.A higher number of culturable fungi were obtained from 13 year-old vines,followed by eight and three yearold vines.In the culture-independent approach,a fungal richness of 59 operational taxonomic units(OTU)was detected,being highest in 13 year-old grapevines,followed by eight and three years.Even though the cultivation approach detected lower fungal richness,the results related to stem are consistent for fungal community composition and richness.Comparison of the fungal taxa identified by the two approaches resulted in an overlap of 53%of the fungal genera.Due to interspecific variability of the sequences from NGS,in many cases the OTUs(even with the highly abundant ones)were only assignable to order,family or genus level.Incorporating multi-gene phylogenies we successfully identified many of the NGS derived OTUs with poor taxonomic information in reference databases to the genus or species levels.Hence,this study signifies the importance of applying both culture-dependent and culture-independent approaches to study the fungal endophytic community composition in Vitis vinifera.This principle could also be applied to other host species and ecosystem level studies.