Over the last two decades the molecular phylogeny and classification of Metarhizium has been widely studied. Despite these efforts to understand this enigmatic genus, the basal lineages in ...Metarhizium are still poorly resolved. In this study, a phylogenetic framework is reconstructed for the Clavicipitaceae focusing on Metarhizium through increased taxon-sampling using five genomic loci (SSU, LSU, tef, rpb1, rpb2) and the barcode marker ITS rDNA. Multi-gene phylogenetic analyses and morphological characterisation of green-spored entomopathogenic Metarhizium isolates from Thailand and soil isolates of M. carneum and M. marquandii reveal their ecological, genetic and species diversity. Nineteen new species are recognised in the Metarhizium clade with narrow host ranges: two new species are found in the M. anisopliae complex – M. clavatum on Coleoptera larvae and M. sulphureum on Lepidoptera larvae; four new species are found in the M. flavoviride complex – M. biotecense and M. fusoideum on brown plant hoppers (Hemiptera), M. culicidarum on mosquitoes, M. nornnoi on Lepidoptera larvae; three new species M. megapomponiae, M. cicadae, M. niveum occur on cicadas; five new species M. candelabrum, M. cercopidarum, M. ellipsoideum, M. huainamdangense M. ovoidosporum occur on planthoppers, leafhoppers and froghoppers (Hemiptera); one new species M. eburneum on Lepidoptera pupae; and four new species M. phuwiangense, M. purpureum, M. purpureonigrum, M. flavum on Coleoptera. Of these 19 new species, seven produce a sexual morph (M. clavatum, M. eburneum, M. flavum, M. phuwiangense, M. purpureonigrum, M. purpureum, and M. sulphureum) and asexual morphs are found in the remaining new species and also in M. sulphureum, M. purpureonigrum and M. purpureum. Metarhizium blattodeae, M. koreanum and M. viridulum are new records for Thailand. An alternative neotype for Metarhizium anisopliae is proposed based on multi-gene and 5′tef analyses showing that CBS 130.71 from Ukraine is more suitable, being from a much closer geographical location to Metchnikoff’s Metarhizium anisopliae. This isolate is distinct from the neotype of Metarhizium anisopliae var. anisopliae proposed by M. Tulloch from Ethiopia (ARSEF 7487). Six new genera are established for monophyletic clades subtending the core Metarhizium clade, including Keithomyces, Marquandomyces, Papiliomyces, Purpureomyces, Sungia, and Yosiokobayasia. Metarhizium carneum, M. aciculare, and M. neogunnii are combined in Keithomyces and one new combination for M. marquandii in Marquandomyces is proposed. Purpureomyces is introduced for species producing purple stromata including a new combination for M. khaoyaiense and two new species P. maesotensis and P. pyriformis. Papiliomyces contains two new combinations for M. liangshanense and Metacordyceps shibinensis. The genus Sungia is proposed for the Korean species M. yongmunense on Lepidoptera pupa and Yosiokobayasia for the Japanese species M. kusanagiense also on Lepidoptera pupa. A synoptic and dichotomous key to the accepted taxa is provided together with tables listing distinguishing morphological characters between species, host preferences, and geography.
A new genus and eight new species, all with isaria-like phialides, are described in Cordycipitaceae from Thailand. The new genus, Samsoniella, is segregated from Akanthomyces based on morphological ...and molecular evidence. Samsoniella differs from Akanthomyces in producing orange cylindrical to clavate stromata with superficial perithecia and orange conidiophores with isaria-like phialides and white to cream conidia. A new combination for CBS 240.32, originally identified as Paecilomyces farinosus (Isaria farinosa), and CBS 262.58, originally identified as Penicillium alboaurantium, respectively, is made in Samsoniella. Two new species, Samsoniella aurantia and S. inthanonensis, are described from lepidopteran larvae. Two new species of Cordyceps, C. blackwelliae and C. lepidopterorum, were also found on coleopteran and lepidopteran larvae. Both produce isaria-like morphs with globose phialides and attenuated long necks and white mycelium in culture. The authors established a sexual-asexual link for Cordyceps javanica (= Isaria javanica) on lepidopteran larvae. Four new species, Akanthomyces kanyawimiae, A. sulphureus, A. thailandicus, and A. waltergamsii, were pathogenic on spiders, with some strains of A. kanyawimiae also found on unidentified insect larvae. These four species of Akanthomyces occur on the underside of leaves and produce white to cream white powdery conidia, whereas S. aurantia and S. inthanonensis were found in leaf litter and produce bright orange stromata and synnemata with white conidia. Another new combination, Akanthomyces ryukyuensis, is proposed. Phylogenetic analyses based on a combined data set comprising the nuc rDNA region encompassing the internal transcribed spacers 1 and 2 along with the 5.8S rDNA (ITS), nuc 28S rDNA (28S), partial sequences of translation elongation factor 1-α gene (TEF1), and the genes for RNA polymerase II largest (RPB1) and second-largest (RPB2) subunits strongly support the delimitation of these new species of Cordyceps, Akanthomyces, and in a new genus Samsoniella in Cordycipitaceae.
Two new fungal genera and six species occurring on insects in the orders Orthoptera and Phasmatodea (superorder Orthopterida) were discovered that are distributed across three families in the ...Hypocreales. Sixty-seven sequences generated in this study were used in a multi-locus
phylogenetic study comprising SSU, LSU, TEF, RPB1 and RPB2 together with the nuclear intergenic region (IGR). These new taxa are introduced as Metarhizium gryllidicola, M. phasmatodeae, Neotorrubiella chinghridicola, Ophiocordyceps kobayasii,
O. krachonicola and Petchia siamensis. Petchia siamensis shows resemblance to Cordyceps mantidicola by infecting egg cases (ootheca) of praying mantis (Mantidae) and having obovoid perithecial heads but differs in the size of its perithecia and ascospore shape.
Two new species in the Metarhizium cluster belonging to the M. anisopliae complex are described that differ from known species with respect to phialide size, conidia and host. Neotorrubiella chinghridicola resembles Torrubiella in the absence of a stipe and can
be distinguished by the production of whole ascospores, which are not commonly found in Torrubiella (except in Torrubiella hemipterigena, which produces multiseptate, whole ascospores). Ophiocordyceps krachonicola is pathogenic to mole crickets and shows resemblance to
O. nigrella, O. ravenelii and O. barnesii in having darkly pigmented stromata. Ophiocordyceps kobayasii occurs on small crickets, and is the phylogenetic sister species of taxa in the 'sphecocephala' clade.
In the course of our exploration of the Thai invertebrate-pathogenic fungi for biologically active metabolites, pigmentosin A (
) and a new bis(naphtho-α-pyrone) derivative, pigmentosin B (
), were ...isolated from the spider-associated fungus
sp. Furthermore, a new glycosylated asperfuran
, together with one new (
) and two known (
and
) cyclodepsipeptides, was isolated from
. The pigmentosins
and
showed to be active against biofilm formation of
DSM1104. The lack of toxicity toward the studied microorganism and cell lines of pigmentosin B (
), as well as the antimicrobial effect of pigmentosin A (
), made them good candidates for further development for use in combination therapy of infections involving biofilm-forming
. The structure elucidation and determination of the absolute configuration were accomplished using a combination of spectroscopy, including 1D and 2D NMR, HRMS, Mosher ester analysis, and comparison of calculated/experimental ECD spectra. A chemotaxonomic investigation of the secondary metabolite profiles using analytical HPLC coupled with diode array detection and mass spectrometry (HPLC-DAD-MS) revealed that the production of pigmentosin B (
) was apparently specific for
sp., while the glycoasperfuran
was specific for
.
Fungal specimens parasitic on spider egg sacs (
Araneidae sensu lato
) were collected, isolated, and identified based on molecular phylogenetic analyses of five nuclear loci (ITS, LSU,
TEF1
,
RPB1
...and
RPB2
) combined with morphological data. In this study, one novel monotypic genus is described,
Bhushaniella rubra
for Thailand
. Bhushaniella rubra
is characterized by producing superficial perithecia. Its anamorph has a unique character by producing verticillate phialides with a slightly curved neck. A concurrent evaluation of the secondary metabolites of the mycelial extracts of the new fungus revealed the presence of picoline alkaloids of the penicolinate type, for which we propose the trivial names penicolinates F and G. Their chemical structures were elucidated by two-dimensional nuclear magnetic resonance (2D-NMR) spectroscopy and high resolution mass spectrometry (HR-MS). They only showed weak to no antibiotic activity and were devoid of significant cytotoxic effects.
Fungal specimens parasitic on ants producing
Stilbella
-like anamorph were collected from Thailand. Based on morphological characters and phylogenetic analyses, a new record for
Ophiocordyceps ...buquetii
and a new species,
Ophiocordyceps laotii
, were identified.
Ophiocordyceps laotii
is a pathogen specific to the ant species
Dolichoderus laotius
, while
O. buquetii
is a species distributed globally with a broad range of ant host species, occurring on
Camponotus
sp.,
Crematogaster
sp.,
Dilobocondyla
sp
.
,
Oecophylla smaragdina
,
Platythyrea
sp.,
Polyrhachis illaudata
,
Polyrhachis
sp. 1, and
Polyrhachis
sp. 2.
Ophiocordyceps buquetii
and
O. laotii
share the morphological character of producing multiple synnemata from ant hosts but differ in the shape of fertile head and conidia, as well as its phylogenetic placement, which clearly distinguishes these species
.
Nine species in
Cordyceps
sensu
lato
producing orange to red stromata were discovered during diversity surveys of arthropod-pathogenic fungi in Thailand. The phylogenetic analyses of combined ITS, ...LSU,
RPB1
,
RPB2
and
TEF1
sequence data indicated four novel species belonging to
Blackwellomyces
and five novel species of
Cordyceps.
All
Blackwellomyces
species produce filiform ascospores with septations.
Blackwellomyces aurantiacus
and
Blackwellomyces roseostromatus
occur on lepidopteran larvae, and both species produce an
Evlachovaea
-like conidial arrangement of the anamorph and produce a red pigment that diffuses across agar medium.
Blackwellomyces calendulinus
and
Blackwellomyces
minutus
are found on coleopteran larvae producing
Acremonium-
like and
Mariannaea
-like conidial arrangements of their anamorphs and do not produce any pigment. Novel members of
Cordyceps
include the species
Cordyceps brevistroma
,
Cordyceps inthanonensis
,
Cordyceps neopruinosa
, and
Cordyceps parvistroma
that are pathogenic on lepidopteran larvae and cocoons, whereas
C. araneae
is pathogenic on spiders
.
All
Cordyceps
species produce bola-shaped, whole ascospores, except for
C. inthanonensis
that produces ascospores dissociating into part-spores. A common feature of the novel
Cordyceps
species presented herein is the production of
Evlachovaea
-like or
Mariannaea
-like conidial arrangement of the anamorphs.
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•A prenyl-tryptophan dimer was isolated from Conoideocrella luteorostrata.•The structure was elucidated by spectroscopic analysis.•Conoideoxime A displayed antibacterial activity.
...Conoideoxime A (1), a prenyl-tryptophan dimer possessing bis-oxime functionality, was isolated from cultures of the whitefly pathogenic fungus Conoideocrella luteorostrata BCC 76664. The structure was elucidated by spectroscopic analyses. It displayed antibacterial activity against Gram-positive bacteria, Bacillus cereus, Enterococcus faecium, and Staphylococcus aureus, with MIC values of 3.13, 6.25, and 6.25 μg/mL, respectively.
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•Host-specific species of Ophiocordyceps unilateralis s. l. supported by whole-genome data.•Occasional host-shift and hybridization without persistent introgression.•Genetic clusters ...suggest the existence of further cryptic species.•Limited recombinations within species and genetic clusters suggesting high inbreeding.•North/Center vs. South biogeographic genetic structure in Thailand.
The identification and delimitation of species boundaries are essential for understanding speciation and adaptation processes and for the management of biodiversity as well as development for applications. Ophiocordyceps unilateralis sensu lato is a complex of fungal pathogens parasitizing Formicine ants, inducing zombie behaviors in their hosts. Previous taxonomic works with limited numbers of samples and markers led to the “one ant-one fungus” paradigm, resulting in the use of ant species as a proxy for fungal identification. Here, a population genomics study with sampling on three ant species across Thailand supported the existence of host-specific species in O. unilateralis s.l. with no footprints of long term introgression despite occasional host shifts and first-generation hybrids. We further detected genetic clusters within the previously delimited fungal species, with each little footprints of recombination, suggesting high levels of inbreeding. The clusters within each of O. camponoti-leonardi and O. camponoti-saundersi were supported by differentiation throughout the genome, suggesting they may constitute further cryptic species parasitizing the same host, challenging the one ant-one fungus paradigm. These genetic clusters had different geographical ranges, supporting different biogeographic influences between the north/center and the south of Thailand, reinforcing the scenario in which Thailand endured compartmentation during the latest Pleistocene glacial cycles.
Four new species of the genus Niveomyces are described from Thailand. They were found as mycoparasites on: Ophiocordyceps infecting flies ( Diptera ) for Niveomyces albus ; ants ( Hymenoptera ) for ...N. formicidarum ; and leafhoppers ( Hemiptera ) for N. hirsutellae and N. multisynnematus . A new genus, Pseudoniveomyces with two species: Pseudoniveo. blattae (type species), parasitic on Ophiocordyceps infecting cockroaches, and Pseudoniveo. arachnovorum , found on a spider egg sac, are also described. These fungi share a common feature which is a sporothrix-like asexual morph. Based on our molecular data, Sporothrix insectorum is shown to be affiliated to the genus Niveomyces , and thus a new combination N. insectorum comb. nov. is proposed. Niveomyces coronatus , N. formicidarum and N . insectorum formed the N. coronatus species complex found on ant-pathogenic Ophiocordyceps from different continents. Pseudoniveomyces species are distinguished from Niveomyces spp. based on the presence of fusoid macroconidia in culture and a red pigment diffused in the medium, resembling to Gibellula and Hevansia . The molecular phylogenetic analyses also confirmed its generic status. The host/substrates associated with the genera within Cordycipitaceae were mapped onto the phylogeny to demonstrate that mycoparasitism also evolved independently multiple times in this family.
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