Content uploaded by Miroslav Kolarik
Author content
All content in this area was uploaded by Miroslav Kolarik on Apr 04, 2014
Content may be subject to copyright.
A preview of the PDF is not available
New Claviceps species from warm-season grasses
Abstract and Figures
Eight undescribed species of Claviceps were recognized from the combinations of molecular and morphological characters. The teleomorph was observed only for Claviceps setariicola. Phylogenetic affinities of the new species inside the genus were revealed by a 5.8S-ITS-28S nrDNA analysis. Claviceps chloridicola, C. tenuispora, C. setariicola and C. setariiphila are related to C. maximensis; C. truncatispora is a sister species to C. pusilla. Claviceps clavispora and C. langdonii cluster with species colonizing maize and sorghum. The position of C. loudetiae is unclear. Comparisons with herbarium specimens showed C. setariicola as a well-established species on Setaria spp. in the southern USA. C. tenuispora was recorded on Cenchrus and Pennisetum in Brazil, USA, and Zimbabwe. C. setariiphila was found on S. geniculata in Brazil. C. chloridicola, C. loudetiae and C. truncatispora occurred in African savannas on Chloris, Loudetia, and Hyparrhenia spp., respectively. C. clavispora was found on Paspalum sp. and Urochloa sp. in Mexico and C. langdonii colonized Dichanthium spp. in the southern USA and probably in Mexico. The occurrence of C. pusilla on pearl millet in the USA (Texas) is reported and the record of C. sulcata on Urochloa brizantha in Brazil is confirmed by nrDNA sequence comparison with an African herbarium specimen. No alkaloids were detected in sclerotia
and/or sphacelia of the new species.
KeywordsAscomycota–Taxonomy–Phylogeny–
Clavicipitales
–Ergot
Figures - uploaded by Miroslav Kolarik
Author content
All figure content in this area was uploaded by Miroslav Kolarik
Content may be subject to copyright.
... 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. ...
... pusilla" (CCC 499) from Australia. All the species in the C. pusilla clade have similar triangular macroconidia (Pažoutová et al. 2011, van der Linde et al. 2022, thus this type of macroconidia can be considered a morphological synapomorphy. ...
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.
... Claviceps species are the causal agents of the ubiquitous ergot disease of sedges, rushes, and grasses (Pažoutová 2002). Ergot in Paspalum spp. is caused mainly by Claviceps paspali described by Stevens and Hall (1910) in USA and C. clavispora described recently by Pažoutová and Odvody (2011) in Mexico. In addition, C. queenslandica was reported by Langdon (1954) affecting only P. orbiculare in Australia. ...
... Sequences of regions of rDNA (Pažoutová 2001;Spatafora et al. 2007), fragments of the minichromosome maintenance complex component 7 (MCM7) gene (Aguileta et al. 2008), regions of the elongation factor 1-α (TEF1) and β-tubulin (TUB2) genes (Tooley et al. 2001), and part of the RNA polymerase subunit II gene (RPB2) (Píchová et al. 2018) have been used to assess taxonomic relatedness among Claviceps species and to develop identification techniques (Tooley et al. 2001;Gilmore et al. 2016). Randomly amplified polymorphic DNA (RAPD) method has been successfully used to discriminate different Claviceps species (Pažoutová and Parbery 2005) and especially to type C. purpurea isolates (Jungehulsing and Tudzynski 1997;Pažoutová et al. 2000Pažoutová et al. , 2002Fisher et al. 2005;Pažoutová and Odvody 2011;Irzykowska et al. 2012). However, molecular studies of Claviceps species infecting P. dilatatum and P. plicatulum are scarce. ...
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 Japan, S. Tanda investigated Claviceps in a series of papers published from 1977 to 1992 and described five new species and several new varieties. The last significant contribution to ergot taxonomy was made by S. Pažoutová ( † 6th September 2013) and colleagues, who described fifteen ergot species from America and Africa (Pažoutová et al., 1998(Pažoutová et al., , 2008a(Pažoutová et al., , 2011Van der Linde et al. 2016) and studied cryptic species within C. purpurea s.l. (Pažoutová et al., 2000b(Pažoutová et al., , 2015. ...
... EAs, and in particular ergovaline, were found to have no effect on endophytic growth in Epichloë (Panaccione et al., 2001;Mulinti et al., 2016). Numerous Clavicipitaceae species lack alkaloids, which is also true for some C. purpurea individuals (Pažoutová et al., 2011;Supplementary Table A.3), suggesting that EAs are not necessary for pathogenic growth. Nothing is known about ergochromes. ...
The ergot, genus Claviceps, comprises approximately 60 species of specialised ovarial grass parasites famous for the production of food toxins and pharmaceutics. Although the ergot has been known for centuries, its evolution have not been resolved yet. Our approach combining multilocus phylogeny, molecular dating and the study of ecological, morphological and metabolic features shows that Claviceps originated in South America in the Palaeocene on a common ancestor of BEP (subfamilies Bambusoideae, Ehrhartoideae, Pooideae) and PACMAD (subfamilies Panicoideae, Aristidoideae, Chloridoideae, Micrairoideae, Arundinoideae, Danthonioideae) grasses. Four clades described here as sections diverged during the Paleocene and Eocene. Since Claviceps are parasitic fungi with a close relationship with their host plants, their evolution is influenced by interactions with the new hosts, either by the spread to a new continent or the radiation of the host plants. Three of the sections possess very narrow host ranges and biogeographical distributions and have relatively low toxicity. On the contrary, the section Claviceps, comprising the rye ergot, C. purpurea, is unique in all aspects. Fungi in this section of North American origin have spread all over the world and infect grasses in all subfamilies as well as sedges, and it is the only section synthesising toxic ergopeptines and secalonic acids. The evolutionary success of the Claviceps section members can be explained by high toxin presence, serving as feeding deterrents and playing a role in their protective mutualism with host plants. Closely related taxa Neoclaviceps monostipa and Cepsiclava phalaridis were combined into the genus Aciculosporium.
... While many plant-inhabiting fungi are regarded as endophytes in the Clavicipitaceae, some members of this family adopt an epibiotic existence whereby they live superficially on the exterior of host plants (15). Ergot alkaloids, a group of mycotoxins often produced by clavicipitaceous fungi, have been reported from several C4 and C3 grass species (16). For example, ergot alkaloids were detected in situ from three Balansia epichloë-infected grass leaves, including Sporobolus from the southeastern United States (17). ...
Grasses harbor diverse fungi, including some that produce mycotoxins or other secondary metabolites. Recently, Florida cattle farmers reported cattle illness, while the cattle were grazing on warm-season grass pastures, that was not attributable to common causes, such as nutritional imbalances or nitrate toxicity. To understand correlations between grass mycobiome and mycotoxin production, we investigated the mycobiomes associated with five prominent, perennial forage and weed grasses [Paspalum notatum Flügge, Cynodon dactylon (L.) Pers., Paspalum nicorae Parodi, Sporobolus indicus (L.) R. Br., and Andropogon virginicus (L.)] collected from six Florida pastures actively grazed by livestock. Black fungal stromata of Myriogenospora and Balansia were observed on P. notatum and S. indicus leaves and were investigated. High-throughput amplicon sequencing was applied to delineate leaf mycobiomes. Mycotoxins from P. notatum leaves were inspected using liquid chromatography-mass spectrometry (LC-MS/MS). Grass species, cultivars, and geographic localities interactively affected fungal community assemblies of asymptomatic leaves. Among the grass species, the greatest fungal richness was detected in the weed S. indicus. The black fungal structures of P. notatum leaves were dominated by the genus Myriogenospora, while those of S. indicus were codominated by the genus Balansia and a hypermycoparasitic fungus of the genus Clonostachys. When comparing mycotoxins detected in P. notatum leaves with and without M. atramentosa, emodin, an anthraquinone, was the only compound which was significantly different (P < 0.05). Understanding the leaf mycobiome and the mycotoxins it may produce in warm-season grasses has important implications for how these associations lead to secondary metabolite production and their subsequent impact on animal health. IMPORTANCE The leaf mycobiome of forage grasses can have a major impact on their mycotoxin contents of forage and subsequently affect livestock health. Despite the importance of the cattle industry in warm-climate regions, such as Florida, studies have been primarily limited to temperate forage systems. Our study provides a holistic view of leaf fungi considering epibiotic, endophytic, and hypermycoparasitic associations with five perennial, warm-season forage and weed grasses. We highlight that plant identity and geographic location interactively affect leaf fungal community composition. Yeasts appeared to be an overlooked fungal group in healthy forage mycobiomes. Furthermore, we detected high emodin quantities in the leaves of a widely planted forage species (P. notatum) whenever epibiotic fungi occurred. Our study demonstrated the importance of identifying fungal communities, ecological roles, and secondary metabolites in perennial, warm-season grasses and their potential for interfering with livestock health.
... Yet, in the United States alone, C. purpurea can colonize more than 160 different grass species [120]. Ergot of other crops, e.g., sorghum and pearl millet, have a different causal agent that does not contaminate grain with the same alkaloids [121][122][123][124][125][126]. ...
Mycotoxins in small grains are a significant and long-standing problem. These contaminants may be produced by members of several fungal genera, including Alternaria, Aspergillus, Fusarium, Claviceps, and Penicillium. Interventions that limit contamination can be made both pre-harvest and post-harvest. Many problems and strategies to control them and the toxins they produce are similar regardless of the location at which they are employed, while others are more common in some areas than in others. Increased knowledge of host-plant resistance, better agronomic methods, improved fungicide management, and better storage strategies all have application on a global basis. We summarize the major pre- and post-harvest control strategies currently in use. In the area of pre-harvest, these include resistant host lines, fungicides and their application guided by epidemiological models, and multiple cultural practices. In the area of post-harvest, drying, storage, cleaning and sorting, and some end-product processes were the most important at the global level. We also employed the Nominal Group discussion technique to identify and prioritize potential steps forward and to reduce problems associated with human and animal consumption of these grains. Identifying existing and potentially novel mechanisms to effectively manage mycotoxin problems in these grains is essential to ensure the safety of humans and domesticated animals that consume these grains.
Ergot is a fungal disease of many plants but is perhaps most commonly associated with domesticated grasses or cereals, such as rye, wheat, barley, oat, sorghum, millet, maize and rice. Ergot is of historical significance, having been reported for several millennia, but is also of concern in modern agricultural production systems. Caused by many different species within the genus Claviceps , the fungi cause the production of sclerotia, which are typically dark in colour, in place of healthy grain. The sclerotia contain toxins that can make the grain unsafe for consumption by humans or livestock. Ergot can be managed both preharvest as well as postharvest to minimize the presence of sclerotia and their associated toxins in food and feed systems. In this review, we provide a detailed update on our current knowledge of ergot on cereals, with a focus on recent advances in our understanding of fungal toxins and their regulation, pathogen biology and disease management.
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 ergots. 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.
Taxonomic novelties: New species: Claviceps miscanthicola E. Tanaka, Claviceps oplismeni E. Tanaka, Claviceps palustris E. Tanaka, Claviceps phragmitis E. Tanaka, Claviceps sasae E. Tanaka, Claviceps tandae E. Tanaka; New status and combination: Claviceps agropyri (Tanda) E. Tanaka, Claviceps kawatanii (Tanda) E. Tanaka; Typifications (basionyms): Lecto- and epitypification: Claviceps yanagawaensis Togashi; Neotypifications: Claviceps purpurea var. agropyri Tanda, Claviceps bothriochloae Tanda & Y. Muray, Claviceps imperatae Tanda & Kawat., Claviceps microspora var. kawatanii Tanda, Claviceps litoralis Kawat., Claviceps microspora Tanda, Claviceps panicoidearum Tanda & Y. Harada; Resurrection: Claviceps queenslandica Langdon.
Citation: Tanaka E, Tanada K, Hosoe T, Shrestha B, Kolařík M, Liu M (2023). In search of lost ergots: phylogenetic re-evaluation of Claviceps species in Japan and their biogeographic patterns revealed. Studies in Mycology 106: 1–39. doi: 10.3114/sim.2022.106.01
O gênero Cenchrus encontra-se distribuído em todo mundo. No Brasil, a
espécie mais importante para produção animal é a Cenchrus ciliaris L., popularmente
conhecida como Capim-buffel. Esta gramínea se encontra principalmente no
Nordeste, devido a sua alta resistência a seca e ao pastejo. A base de cultivares é
estreita e o cultivar mais difundido é o Biloela. O uso do Capim-buffel no Nordeste
tem permitido aumentar a capacidade de suporte e proporcionar maior desempenho
animal na pastagem nativa.
O objetivo deste capítulo é caracterizar o gênero Cenchrus, com ênfase nos
aspectos de plantio, manejo e utilização do Capim-buffel.
Ergot, the genus Claviceps comprises several deeply diverged lineages, recently classified as sections. Among them, the section Pusillae, is the most speciose, with a centre of distribution in Africa but occurring worldwide, often as a consequence of its invasive potential. This section includes the most severe plant pathogens such as Claviceps africana and C. gigantea, responsible for toxicoses and a significant reduction in the seed yields of Sorghum and Zea. In this study we surveyed ergot diversity in South Africa, focusing on grasses native to this region, but known for their high potential of invasiveness. The revision based on molecular and phenotypic markers revealed 16 species, with a high proportion of undescribed diversity, confirming Africa as a hot spot for this section. Five new species, Claviceps tulasnei, Claviceps eulaliae, Claviceps hypertheliae, Claviceps fredericksoniae and Claviceps arundinellae were described from Setaria, Eulalia, Hyperthelia, Miscanthus and Arundinella respectively. Claviceps texensis infecting Cenchrus, previously only identified from the same host in Texas, USA, was confirmed to be present in Africa, which is assumed to be its primary area of distribution. In addition, the host grass genus Anthephora is newly reported as a host of Claviceps digitariae. The most of the taxa were negligible concerning alkaloid production, with the exception of C. fredericksoniae, which is a sister of potent alkaloid producer C. africana, and produces mainly DH-ergosine, together with traces of DH-ergocornine. The host/parasite associations within Pusillae section is very narrow, suggesting that co-speciation is the major speciation driver in this group. Host grasses of the described species are already recognised invasive species and their ovarial parasites need to be monitored. This is highlighted by the fact that all Pusillae produced air-borne secondary conidia, which is autapomorphy of this section and considered to be important for their invasive abilities.
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.
Isolates of Claviceps species with lunate to fusiform macroconidia were collected from panicoid grasses in Texas and Zimbabwe and described as new species based on anamorphs since no teleomorphs were available. Characterization was based upon morphology and partial sequences of rDNA and beta-tubulin. The isolates grouped into two strongly-supported clades. The first clade contained ancestral C. hirtella and C. fusiformis from pearl millet (Pennisetum glaucum) in clade terminal position with Texas isolates from native cup grass (Eriochloa sericea) and pearl millet grouped between them. The second clade consisted of African isolates from Urochloa and Eragrostis. The isolates from Texas from pearl millet and buffel grass (Cenchrus ciliaris) and isolates from E. sericea were described as new species, Sphacelia texensis and Sphacelia eriochloae, respectively. Both species had morphology, DNA markers, and alkaloid production that was intermediate between those features exhibited in C. fusiformis and C. hirtella. The African isolates from Urochloa and Eragrostis were also described as a new species, Sphacelia lovelessii. In shaken cultures, C. hirtella readily produced a whole range of clavines with agroclavine and festuclavine predominating, but ergometrine was also detected. Claviceps fusiformis produced mainly agroclavine and elymoclavine, S. eriochloae produced mainly agroclavine, elymoclavin and festuclavine and the cultures of S. texensis contained small amounts of agroclavine and festuclavine. Only traces of clavines were found in cultures of S. lovelessii of the second clade. The alkaloid content of infected florets in the sphacelial (honeydew) developmental stage was also measured. Only C. fusiformis and S. eriochloae produced alkaloids in planta at this early stage.
In our previous study of Claviceps purpurea three populations were found: G1 on open localities, G2 from shady or wet habitats and G3 on Spartina stands of coastal salt marshes. The latter two are also chemoraces. In the Czech Republic, isolates of G1 and G2 were found. The ability of four isolates representing these populations to infect and develop sclerotia on different host species (Holcus lanatus, Helictotrichon pubescens, Phalaris arundinacea, Dactylis glomerata, Arrhenatherum elatius, Bromus inermis, Bromus erectus, Elytrigia repens, Avenella flexuosa, Lolium perenne, Poa nemoralis, Poa annua, and different cultivars of Poa pratensis) was studied along with their alkaloid production. P. pratensis and D. glomerata were infected by all the isolates and sclerotia were formed by isolates 207 (G1) and 434 (G2), and on two P. pratensis cultivars even by 481 (G3). Infection ability (formation of sphacelial stage and honeydew) was less host-restricted than formation of mature sclerotia. G2 and G3 strains infected A. flexuosa without sclerotia formation. L. perenne was infected only once by strain 207 (G1) without sclerotia formation. P. annua (natural host of G2), was infected by all isolates, but no sclerotia were formed even with G2 strains. From the two G2 isolates, strain 434 from Dactylis formed sclerotia on five host species, whereas isolate 475 originating from Phragmites stand formed only sphacelia. Composition of alkaloid mixture produced in sclerotia of the same strain from various hosts confirmed that host plant does not influence the type of alkaloids produced, only their ratio.
Previous investigators have identified a number of ergot alkaloids (EAs) in tall fescue (Festuca arundinacea Schreb.) infected by the endophytic fungus, Neotyphoidium coenophialum [(Morgan-Jones & W. Gams) Glenn, Bacon & Hanlin comb. nov.]. Their results, however, may have been confounded by the presence of the related parasitic ergot fungus (Claviceps purpurea [Fr.:Fr.] Tul.), which also produces EAs. Semipreparative HPLC was used to separate fractions giving a high fluorescence response in a sample of endophyte-infected (EI) tall fescue seeds, carefully examined to eliminate the possibility of Claviceps infection. Analytical high-performance liquid chromatography (HPLC) and LC/electrospray ionization mass spectrometry (ESI-MS) were used to identify EAs after isolation. Clearly identified in the spectra were ergine, ergovaline, ergosine, ergonine, and previously undescribed EAs, didehydroergovaline and aci-ergovaline, including their epimers. Several additional EAs may have been present, but structures could not be confirmed by their spectra. Not found in the isolated fractions were ergonovine, ergotamine, or lysergylmethylcarbinolamide. The developed HPLC method was used to determine the alkaloids in plants, culms, and seeds of EI tall fescue.