Fig 5 - uploaded by Miroslav Kolarik
Content may be subject to copyright.
e C. humidiphila. (A) CCC434, culture on T2 medium after 2 weeks AT 25 C. (B) PRM922709 conidia from Sphacelia morph. (C) PRM922709 sclerotia. Scale bar [ 10 mm.
Similar publications
Vier Sunius-Arten werden beschrieben und abgebildet: S. tronqueti sp. n. (Spanien: Sierra Nevada), S. filabresicus sp. n. (Spanien: Sierra de los Filabres), S. gadoricus sp. n. (Spanien: Sierra de Gádor) und S. kastcheevi sp. n. (Kasachstan). Zwei Namen werden synonymisiert: S. melanocephalus (Fabricius, 1793) = S. anatolicus Assing, 1995, syn. n.,...
Citations
... However, the centre of Claviceps diversity is presumed to be in tropical and temperate regions (Píchová et al. 2018). Multi-locus phylogenetic analysis has recently advanced our understanding of species delimitation in Claviceps (Pažoutová et al. 2011(Pažoutová et al. , 2015Liu et al. 2020, van der Linde et al. 2022. By 2000, eight species and six varieties of Claviceps had been described based on Japanese specimens, i.e. C. amamiensis on Digitaria setigera (Tanda 1992a), C. bothriochloae on Capillipedium parviflorum (Tanda 1991e), C. imperatae on Imperata cylindrica (Tanda & Kawatani 1976), C. litoralis on Leymus mollis (Kawatani 1946), C. microspora on Arundinella hirta (Tanda 1991d), C. panicoidearum on Isachne globosa (Tanda & Harada 1989), C. sorghicola on Sorghum bicolor (Tsukiboshi et al. 1999b), C. yanagawaensis on Zoysia japonica (Togashi 1936) and the varieties C. microspora var. ...
... In recent years, molecular studies have changed the status of three varieties. Pažoutová et al. (2015) proposed Claviceps humidiphila based on the description of C. purpurea var. phalaridis from Japan. ...
... Claviceps agropyri (Tanda) Description supplement to the original description by Tanda (1981a) Notes: Tanda (1981a) validly published this taxon as a variety of C. purpurea because its somewhat fusiform-shaped conidial features differed from C. purpurea. Pažoutová et al. (2015) considered this taxon as a synonym of C. purpurea. Our phylogenetic analysis revealed this taxon as a distinct species. ...
Claviceps ( Clavicipitaceae , Hypocreales ) was erected in 1853, although ergotism had been well-known for a much longer time. By 2000, about 70 taxa had been described in Claviceps , of which eight species and six varieties were based on Japanese type or authentic specimens. Most of these Japanese Claviceps taxa are based on lost specimens or have invalid names, which means many species practically exist only in the scientific literature. The ambiguous identities of these species have hindered taxonomic resolution of the genus Claviceps . Consequently, we sought and collected more than 300 fresh specimens in search of the lost Japanese ergot. Multilocus phylogenetic analyses based on DNA sequences from LSU, TEF-1α , TUB2 , Mcm7 , and RPB2 revealed the phylogenetic relationships between the Japanese specimens and known Claviceps spp., as well as the presence of biogeographic patterns. Based on the phylogenetic analysis, host range and morphology, we re-evaluated Japanese Claviceps and recognised at least 21 species in Japan. Here we characterised 14 previously described taxa and designated neo-, lecto- and epi-types for C. bothriochloae , C. imperatae , C. litoralis , C. microspora , C. panicoidearum and C. yanagawaensis . Two varieties were elevated to species rank with designated neotypes, i.e. C. agropyri and C. kawatanii . Six new species, C. miscanthicola , C. oplismeni , C. palustris , C. phragmitis , C. sasae and C. tandae were proposed and described.
... Kolařík. In addition, the new lineage G2a was named C. arundinis (Pažoutová et al. 2015). In terms of the host range and morphology, C. arundinis corresponds well with Tulasne's concept of C. microcephala (Wallr.) ...
... Poa, Dactylis) than others, whereas in particular areas (Central Europe vs Scandinavia) host specificity is more evident at a certain level, such that cultivated cereals in Europe are exclusively infected by C. purpurea s.str. (Pažoutová et al. 2015, Negård et al. 2015. Pažoutová et al. (2000) observed that C. purpurea s.str. ...
... The low resolution at the species level is another shortcoming from which the ITS region in Claviceps spp. suffers; for instance, it could neither separate C. ripicola (G6, 7) from C. spartinae (G3), nor C. arundinis (G2a), C. humidiphila (G2) and C. perihumidiphila (G2b) (Negård et al. 2015;Pažoutová et al. 2015;Shoukouhi et al. 2019). In contrast, the protein-coding genes provide better resolutions, although, for certain species, a fragment of the second largest subunit of RNA polymerase II (RPB2) and the translation elongation factor 1-α (EF1-α) displayed a varied pattern of affinities among species. ...
Ergot fungi produce toxic sclerotia that have significant impacts on the agricultural, food and pharmaceutical industries. These fungi were classified in various genera (such as Sclerotium clavus, or Spermoedia clavus) before Tulasne (1853) erected Claviceps and described three species based on variations in morphology and host ranges, viz. C. purpurea, C. microcephala, and C. nigricans. Since then, knowledge regarding the biological, clinical, and pharmaceutical perspectives of ergot has accumulated rapidly. However, a serious taxonomic examination was lacking until Langdon’s (1952) revision accepted 25 species. That was followed by intensive regional studies by Loveless in the 1960s for Rhodesia (now Zimbabwe, Africa), and Tanda in Japan (1970s – 1990s). More species names were reported and currently over 90 named taxa (species, varieties) are recorded in fungal name repositories (Mycobank and Index Fungorum). Most species were described based on morphological characteristics (sclerotia, ascomata and conidia) and host ranges. Some were characterized by their alkaloid profiles. Recently, DNA multi-locus sequence analyses (MLSA) were applied to resolve species complexes. For instance, the C. purpurea complex was separated into four species, and additional new species were recognized from South Africa and Canada. Infra- and supra-specific level genetic variations were identified via multi-locus and genomic studies. Based on five-locus phylogenies, Píchová and colleagues separated Claviceps into four sections: C. sect. Claviceps, C. sect. Citrinae, C. sect. Paspalorum and C. sect. Pusillae, for 60 species. Among these sections, several doubtful species names require clarification. Careful research on type specimens combined with molecular analyses is essential for clarifying these names.
... The first published record seems to be from Dennis (1975). Claviceps purpurea was considered to be a single species, but in common with many fungi, recent work analysing regions of its DNA demonstrate that it is a complex of several closely related species, some more or less host specific, and some with a wide host range (Pažoutová et al. 2015). ...
... A 2004 study (Alderman et al. 2004) found that C. purpurea infected 165 grass species in the continental United States and Alaska alone. Although C. purpurea is historically considered a cosmopolitan fungus with a broad host range, a series of studies revealed at least three genetic lineages that reflected ecological differentiation and adaptation (Douhan et al. 2008;Pažoutová et al. 2000Pažoutová et al. , 2015. The three lineages were referred to as G1, G2, and G3 ecotypes and were differentiated based on conidial morphology, alkaloid profiles, random amplified polymorphic DNA (RAPD) markers, DNA sequences, and the ability or inability of the sclerotia to float (Jungehülsing and Tudzynski 1997;Pažoutová et al. 2000Pažoutová et al. , 2015. ...
... Although C. purpurea is historically considered a cosmopolitan fungus with a broad host range, a series of studies revealed at least three genetic lineages that reflected ecological differentiation and adaptation (Douhan et al. 2008;Pažoutová et al. 2000Pažoutová et al. , 2015. The three lineages were referred to as G1, G2, and G3 ecotypes and were differentiated based on conidial morphology, alkaloid profiles, random amplified polymorphic DNA (RAPD) markers, DNA sequences, and the ability or inability of the sclerotia to float (Jungehülsing and Tudzynski 1997;Pažoutová et al. 2000Pažoutová et al. , 2015. Recently, C. purpurea ecotypes G1, G2, and G3 were described as three separate species: C. purpurea sensu stricto, C. humidiphila, and C. spartinae, respectively . ...
... purpurea sensu stricto) or G2 ecotype (C. humidiphila), respectively, as described by Pažoutová et al. (2000Pažoutová et al. ( , 2015. b NA, not amplified. ...
Ergot, caused by Claviceps purpurea sensu lato, is an economically important seed replacement disease of Kentucky bluegrass (Poa pratensis) and perennial ryegrass (Lolium perenne) seed crops. Claviceps purpurea sensu stricto is considered the primary Claviceps species responsible, but genetic diversity and cryptic species within C. purpurea sensu lato have previously been reported. Fifty-six C. purpurea sensu lato isolates collected from P. pratensis (n=21) and L. perenne (n=35) in Oregon and Washington between 2010 and 2014 were characterized using RAPD, partial ITS, β-tubulin and elongation factor-1α sequences, conidial size, and ergot alkaloid chemotype. Based on RAPD analysis, 7 isolates from P. pratensis and 33 isolates from L. perenne collected in Oregon corresponded to C. purpurea sensu stricto, while 13 isolates collected from P. pratensis in Washington and Oregon were identified as C. humidiphila. Partial ITS, β-tubulin, and elongation factor-1α sequences identified 10 isolates from P. pratensis as C. humidiphila, while seven isolates from P. pratensis and 33 isolates from L. perenne were identified as C. purpurea sensu stricto. Several isolates generated ambiguous RAPD bands and/or sequences that prevented identification. Ergot alkaloid chemotype profiling found that ergocornine and its epimer were predominant in sclerotia from P. pratensis, whereas ergotamine and its epimer were found in highest abundance in sclerotia from L. perenne. This study confirms the presence of the C. purpurea sensu lato species complex in the U.S. Pacific Northwest and suggests that more research is needed to characterize and mitigate Claviceps spp. infection of grass seed crops in North America.
... A previous investigation of the genetic diversity of Canadian ergot fungi isolated from agricultural areas, using a multilocus sequence analysis (MLSA), revealed seven lineages within the premolecular concept of C. pupurea (Shoukouhi et al. 2019). Three of the seven lineages corresponded to C. humidiphila, C. spartinae, and C. purpurea sensu stricto (Pažoutová et al. 2015), whereas the remaining four lineages did not affiliate with any formally described species (Shoukouhi et al. 2019). Inclusion of two single-copy ergot alkaloid synthesis genes (easA, encoding chanoclavine I aldehyde oxidoreductase, and easE, encoding chanoclavine I synthase oxidoreductase) in the MLSA proved efficacious, due to the presence of a large number of informative characters when compared among species (Shoukouhi et al. 2019). ...
... Conidia from sclerotia and T2 medium were smaller compared with the other three species and C. humidiphila, which was featured as producing larger conidia than C. purpurea on T2. The shapes of conidia from culture were variable, predominantly ellipsoid, pyriform, broad lancinate, or reniform, but seldom oblongoid as in C. humidiphila (Pažoutová et al. 2015). Conidiogenous cells from in vitro culture occur as various shapes: phialidic, cylindrical, or as undifferentiate hyphal cells. ...
... Occasionally bifurcate conidiophores were observed. The interior of sclerotia was pale yellow to grayish yellow, not white-colored as in C. humidiphila (Pažoutová et al. 2015 Sclerotia and stromata were not observed. Only living culture available from historical collections. ...
Four ergot species (Claviceps ripicola, C. quebecensis, C. perihumidiphila, and C. occidentalis) were recognized based on analyses of DNA sequences from multiple loci, including two housekeeping genes, RNA polymerase II second largest subunit (RPB2), and translation elongation factor 1-α (TEF1-α), and a single-copy ergot alkaloid synthesis gene (easE) encoding chanoclavine I synthase oxidoreductase. Morphological features, ergot alkaloid production, and pathogenicity on five common cereal crops of each species were evaluated and presented in taxonomic descriptions. A synoptic key was also provided for identification.
... Since section Paspalorum comprises only C. paspali ( Píchová et al. 2018), the position of the Claviceps isolates from P. plicatulum suggests that they could be considered an ecological subspecies or specialized variants adapted to colonize other Paspalum species, such as P. plicatulum. This situation could mirror the speciation process recently described for C. purpurea cryptic species ( Douhan et al. 2008;Pažoutová et al. 2015). The remarkable separation of the three isolates obtained from P. plicatulum from those obtained from P. dilatatum was confirmed by the four single-gene phylogenetic analyses, by RAPD fingerprinting combined with the presence of EAS and IDT genes, and the observation that no sequence-based haplotype was shared, for any locus, among isolates from both grasses. ...
Claviceps species affecting Paspalum spp. are a serious problem, as they infect forage grasses such as Paspalum dilatatum and P. plicatulum, producing the ergot disease. The ascomycete C. paspali is known to be the pathogen responsible for this disease in both grasses. This fungus produces alkaloids, including ergot alkaloids and indole-diterpenes, that have potent neurotropic activities in mammals. A total of 32 isolates from Uruguay were obtained from infected P. dilatatum and P. plicatulum. Isolates were phylogenetically identified using partial sequences of the genes coding for the second largest subunit of RNA polymerase subunit II (RPB2), translation elongation factor 1-α (TEF1), β-tubulin (TUB2), and the nuc rDNA 28S subunit (28S). Isolates were also genotyped by randomly amplified polymorphic DNA (RAPD) and presence of genes within the ergot alkaloid (EAS) and indole-diterpene (IDT) biosynthetic gene clusters. This study represents the first genetic characterization of several isolates of C. paspali. The results from this study provide insight into the genetic and genotypic diversity of Claviceps paspali present in P. dilatatum and suggest that isolates from P. plicatulum could be considered an ecological subspecies or specialized variant of C. paspali. Some of these isolates show hypothetical alkaloid genotypes never reported before.
... In light of phylogenetic analyses of TEF1-α gene (Fig. 15A), C. zizaniae is grouped in Claviceps sect. Claviceps (Pažoutová et al. 2015;Píchová et al. 2018), however without marked affinity to any species. Phylogenetic analysis based on the ergot alkaloid biosynthesis gene easE suggests that C. zizaniae is more closely related with C. (Fig. 15B). ...
... Multigene studies have confirmed cryptic speciation within C. purpurea, and three infraspecific groups correlated with ecological niches were designated as lineages G1-3 (Pažoutová et al. 2000;Douhan et al. 2008). Pažoutová et al. (2015) discovered a fourth lineage closely related to G2, which they labeled as G2a, and meanwhile recognized the four phylogenetic lineages as species, i.e., C. purpurea (G1), C. humidiphila (G2), C. arundinis (G2a), and C. spartinae (G3). These results were further confirmed with a comprehensive study of Norweigian samples (Negård et al. 2015). ...
... A portion of the nuclear gene encoding translation elongation factor 1-α (TEF1) was amplified and sequenced using EF1-983F and EF1-2218R (Rehner and Buckley 2005). A portion of the second largest subunit of RNA polymerase II (RPB2) was amplified and sequenced using a modified fRPB2-5F specific for ergot fungi (5′-TTTCGTGGTATTGTTCG-CAGA-3′) (Pažoutová et al. 2015) and fRPB2-7cR (Liu et al. 1999). The gene encoding β-tubulin (tubB) was amplified using T1 and T22 and sequenced with primers T1, T2, T12, and T22 (O'Donnell and Cigelnik 1997). ...
... Douhan et al. (2008) confirmed that the separation of three groups (G1-3) correlated with habitats using ITS, an RAS-like locus, and partial tubB. Pažoutová et al. (2015) recognized four phylogenetic species using genealogical concordance phylogenetic analysis based on six loci. Van der Linde et al. (2016) discovered four more phylogenetic species from Africa. ...
The ergot disease of cereals has become increasingly important in agricultural areas of Canada since 1999. Generally, this disease is considered to be caused by Claviceps purpurea, but the taxonomy of Claviceps from these areas has not been well studied. The objectives of this study were (i) to determine the phylogenetic lineages (phylogenetic species) present in agricultural areas of Canada and (ii) to develop a molecular assay that can separate the lineages on crops from other lineages. Genetic diversity of Claviceps collected from agriculture areas in Canada were investigated using multilocus sequence typing. The loci sequenced include nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS), partial fragments of translation elongation factor 1-α (TEF1), RNA polymerase II second largest subunit (RPB2), β-tubulin (tubB), and two ergot alkaloid synthesis genes (easA, easE). Based on individual locus and concatenated alignments, phylogenetic analyses revealed seven lineages within the premolecular concept of C. purpurea, of which five corresponded with undescribed species (G2b and G4-7). Although lineages G2-7 had narrow host ranges, lineage G1 (= C. purpurea s.s.) had a broad host range that overlapped with other lineages. A molecular diagnostic quantitative polymerase chain reaction (qPCR) assay was developed and validated with 185 samples from a wide range of host plants and geographic origins, including 10 phylogenetic species in C. sect. Claviceps, 8 in C. sect. Pusillae, 1 in C. sect. Citrinae, and 1-2 species from Alternaria, Fusarium, and Penicillium. The assay can detect lineage G1 at a concentration of 7.5 pg/μL and distinguish it from other Claviceps species and lineages. This facilitates disease management by detecting the inocula from nonagriculture host plants.
... We considered commending the adoption of the suffix 'agg.' for material when precise molecular species identifications cannot be made. While this has been done in a few other groups of fungi (e.g., Parnmen et al. 2013, Pažoutová et al. 2015, 'complex' has come to be used more widely and was strongly favoured at the Cryptic Speciation in Fungi symposium in Utrecht in September 2017 (report awaited). We therefore suggest the use of 'complex' here but recognise some may prefer to use 'agg.' or 's.lat.'. ...
In many lichen-forming fungi, molecular phylogenetic analyses lead to the discovery of cryptic species within traditional morphospecies. However, in some cases, molecular sequence data also questions the separation of phenotypically characterised species. Here we apply an integrative taxonomy approach ‒ including morphological, chemical, molecular, and distributional characters ‒ to re-assess species boundaries in a traditionally speciose group of hair lichens, Bryoria sect. Implexae. We sampled multilocus sequence and microsatellite data from 142 specimens from a broad intercontinental distribution. Molecular data included DNA sequences of the standard fungal markers ITS, IGS, GAPDH, two newly tested loci (FRBi15 and FRBi16), and SSR frequencies from 18 microsatellite markers. Datasets were analysed with Bayesian and maximum likelihood phylogenetic reconstruction, phenogram reconstruction, STRUCTURE Bayesian clustering, principal coordinate analysis, haplotype network, and several different species delimitation analyses (ABGD, PTP, GMYC, and DISSECT). Additionally, past population demography and divergence times are estimated. The different approaches to species recognition do not support the monophyly of the 11 currently accepted morphospecies, and rather suggest the reduction of these to four phylogenetic species. Moreover, three of these are relatively recent in origin and cryptic, including phenotypically and chemically variable
specimens. Issues regarding the integration of an evolutionary perspective into taxonomic conclusions in species complexes, which have undergone recent diversification, are discussed. The four accepted species, all epitypified by sequenced material, are Bryoria fuscescens, B. glabra, B. kockiana, and B. pseudofuscescens. Ten species rank names are reduced to synonymy. In the absence of molecular data, they can be recorded as the B. fuscescens
complex. Intraspecific phenotype plasticity and factors affecting the speciation of different morphospecies in this group of Bryoria are outlined.
... A SYBR Green qPCR assay was developed for the detection and quantification of C. purpurea sensu lato, including C. purpurea sensu stricto and C. humidiphila, both of which can cause ergot in grass seed production systems of the Pacific Northwest. The assay could also be used to detect and quantify C. spartinae, a pathogen of Spartina and Distichlis grass species in coastal salt marshes (Duncan et al. 2002;Pažoutová et al. 2000Pažoutová et al. , 2015. ...
The U.S. Pacific Northwest states of Oregon and Washington are major producers of cool-season grass seed. Ergot, caused by fungi in the Claviceps purpurea sensu lato group, is an important seed replacement disease of grass worldwide. Microscopic methods that are currently used to quantify airborne Claviceps ascospores captured by spore traps are not currently rapid enough to allow for detecting and reporting of spore numbers in a timely manner, hindering growers from using this information to help manage ergot. We developed a SYBR Green real-time quantitative polymerase chain reaction (qPCR)-based assay for fast and efficient detection and quantification of C. purpurea sensu lato ascospores from Hirst-type spore traps. Species-specificity of the qPCR assay was confirmed against 41 C. purpurea sensu lato isolates collected from six hosts and six other Claviceps spp. Significant relationships were observed between cycle threshold (Ct) values and standard curves of serial dilutions of DNA ranging from 1 pg to 10 ng (R2 = -0.99; P = 0.0002) and DNA extracted from a conidial suspension representing 8 to 80,000 conidia (R2 = -0.99; P = 0.0004). Ct values from qPCR were significantly correlated with results from microscopic examination of spore trap samples from the field (r = -0.68; P < 0.0001) and the procedure was able to detect a single ascospore from spore trap tape samples. The qPCR procedure developed in this study provided a means for quantifying airborne Claviceps ascospores that was highly specific and useful over a wide range of spore densities, and could be performed in a matter of hours instead of days. The qPCR assay developed in this study could be part of an integrated pest management approach to help grass seed growers make risk-based fungicide application decisions for ergot management in grass grown for seed.
