Ducellierialesord. nov. and evidence for a novel clade of endobiotic pollen-infecting "lagenidiaceous" Peronosporomycetes.

The genus Ducellieria (Ducellieriaceae) contains three species (D. chodatii, D. tricuspidata, D. corcontica), and a single variety (D. chodatii var. armata) of obligate endobiotic pollen parasites. These organisms have been first assigned to the green alga genus Coelastrum, as they form very similar spherical structures, but the observation of heterokont zoospores has led to their reclassification to the phylum Oomycota. However, despite their widespread nature, these organisms are only known from their descriptive morphology, and life cycle traits of some species still remain incompletely known. Only the type species, D. chodatii, has been rediscovered several times, but the phylogeny of the genus remains unresolved, since none of its species has been studied for their molecular phylogeny. At present the genus is still included in some algal databases. To clarify the evolutionary affiliation of Ducellieria, efforts were undertaken to isolate D. chodatii from pollen grains, to infer its phylogenetic placement based on nrSSU sequences. By targeted isolation, the pollen endoparasitoid was rediscovered from three lakes in Germany (Mummelsee, Okertalsperre, Knappensee). Apart from the typical coelastrum-like spheroids, oomycetes sporulating directly from pollen grains in a lagenidium-like fashion were observed, and molecular sequences of both types of oomycetes were obtained. Phylogenetic reconstruction revealed that coelastrum-like and lagenidium-like forms are unrelated, with the former embedded within the deep branching early-diverging lineages, and the later stage forming a distinct clade in Peronosporales. Consequently, the life cycle of D. chodatii needs careful revision using single-spore isolates of the species, to infer if previous lifecycle reconstructions that involve various different thallus types are stages of a single species or potentially of several ones. Citation: Buaya AT, Thines M (2023). Ducellieriales ord. nov. and evidence for a novel clade of endobiotic pollen-infecting "lagenidiaceous" Peronosporomycetes. Fungal Systematics and Evolution 12: 247-254. doi: 10.3114/fuse.2023.12.12.

Adding a missing piece to the puzzle of oomycete phylogeny: the placement of Rhipidium interruptum (Rhipidiaceae).

Oomycetes are a group of fungus-like organisms, which phylogenetically comprise early diverging lineages that are mostly holocarpic, and two crown classes, the Peronosporomycetes and Saprolegniomycetes, including many well-investigated pathogens of plants and animals. However, there is a poorly studied group, the Rhipidiales, which placement amongst the crown oomycetes is ambiguous. It accommodates several taxa with a sophisticated vegetative and reproductive cycle, as well as structural organisation, that is arguably the most complex in the oomycete lineage. Despite the remarkable morphological complexity and their notable perseverance in the face of faster-growing saprotrophic oomycetes and fungi, the knowledge on Rhipidiales is limited to date, as the most complex members are not easily cultured, even by targeted approaches. This also leads to inadequate sequence data for the order, which was sourced from only the two least complex out of seven introduced genera, i.e. Sapromyces and Salispina. In the present study, ex-situ baiting was done using various fruit substrates, and naturally-shed twigs or fruits acquired from water bodies were examined. As a result of these efforts, the species Rhipidium interruptum was obtained and gross cultivation was accomplished using poplar (Populus nigra) twigs as substrate, which allowed further documentation of both asexual and sexual reproduction. This enabled phylogenetic and detailed morphological study, as well as an epitypification of the species. Phylogenetic analyses based on cox2 and nrLSU sequences revealed Rhipidium as the sister genus of Sapromyces. The morphological studies done support a conspecificity of R. interruptum and R. continuum, which might in turn be conspecific with R. americanum. Though several further studies will be required to fit the scattered missing pieces of knowledge on Rhipidiales together revealing a more complete picture of oomycete evolution, we hope that the current study can serve as a cornerstone for future investigations in the group. Citation: Tsai I, Thines M (2023). Adding a missing piece to the puzzle of oomycete phylogeny: the placement of Rhipidium interruptum (Rhipidiaceae). Fungal Systematics and Evolution 11: 95-108. doi: 10.3114/fuse.2023.11.08.

Peronosporaceae species causing downy mildew diseases of Poaceae, including nomenclature revisions and diagnostic resources.

Downy mildew pathogens of graminicolous hosts (Poaceae) are members of eight morphologically and phylogenetically distinct genera in the Peronosporaceae (Oomycota, Peronosporales). Graminicolous downy mildews (GDMs) cause severe losses in crops such as maize, millets, sorghum, and sugarcane in many parts of the world, especially in tropical climates. In countries where the most destructive GDMs are not endemic, these organisms are often designated as high-risk foreign pathogens and subject to oversight and quarantine by regulatory officials. Thus, there is a need to reliably and accurately identify the causal organisms. This paper provides an overview of the Peronosporaceae species causing graminicolous downy mildew diseases, with a description of their impact on agriculture and the environment, along with brief summaries of the nomenclatural and taxonomic issues surrounding these taxa. Key diagnostic characters are summarized, including DNA sequence data for types and/or voucher specimens, morphological features, and new illustrations. New sequence data for cox2 and 28S rDNA markers are provided from the type specimens of three species, Peronosclerospora philippinensis, Sclerospora iseilematis, and Sclerospora northii. Thirty-nine species of graminicolous downy mildews are accepted, and seven previously invalidly published taxa are validated. Fifty-five specimens are formally designated as types, including lectotypification of 10 species, neotypification of three species, and holotype designation for Sclerophthora cryophila. Citation: Crouch JA, Davis WJ, Shishkoff N, Castroagudín VL, Martin F, Michelmore R, Thines M (2022). Peronosporaceae species causing downy mildew diseases of Poaceae, including nomenclature revisions and diagnostic resources. Fungal Systematics and Evolution 9: 43-86. doi: 10.3114/fuse.2022.09.05.

High Nucleotide Substitution Rates Associated with Retrotransposon Proliferation Drive Dynamic Secretome Evolution in Smut Pathogens.

Transposable elements (TEs) play a pivotal role in shaping diversity in eukaryotic genomes. The covered smut pathogen on barley, Ustilago hordei, encountered a recent genome expansion. Using long reads, we assembled genomes of 6 U. hordei strains and 3 sister species, to study this genome expansion. We found that larger genome sizes can mainly be attributed to a higher genome fraction of long terminal repeat retrotransposons (LTR-RTs). In the studied smut genomes, LTR-RTs fractions are the largest in U. hordei and are positively correlated with the mating-type locus sizes, which is up to ~560 kb in U. hordei. Furthermore, LTR-RTs were found to be associated with higher nucleotide substitution levels, as these occur in specific genome regions of smut species with a recent LTR-RT proliferation. Moreover, genes in genome regions with higher nucleotide substitution levels generally reside closer to LTR-RTs than other genome regions. Genome regions with many nucleotide substitutions encountered an especially high fraction of CG substitutions, which is not observed for LTR-RT sequences. The high nucleotide substitution levels particularly accelerate the evolution of secretome genes, as their more accessory nature results in substitutions that often lead to amino acid alterations. IMPORTANCE Genomic alteration can be generated through various means, in which transposable elements (TEs) can play a pivotal role. Their mobility causes mutagenesis in itself and can disrupt the function of the sequences they insert into. They also impact genome evolution as their repetitive nature facilitates nonhomologous recombination. Furthermore, TEs have been linked to specific epigenetic genome organizations. We report a recent TE proliferation in the genome of the barley covered smut fungus, Ustilago hordei. This proliferation is associated with a distinct nucleotide substitution regime that has a higher rate and a higher fraction of CG substitutions. This different regime shapes the evolution of genes in subjected genome regions. We hypothesize that TEs may influence the error-rate of DNA polymerase in a hitherto unknown fashion.

Miracula blauvikensis: a new species of Miracula from Iceland, and report of a co-cultivation system for studying oomycete-diatom interactions.

The genus Miracula represents an early-diverging lineage of diatom-parasitic Oomycota, straminipilous eukaryotes that have evolved fungal features independent from the opisthokont Fungi. Recent studies have revealed that diatom parasitoids are much more species-rich than previously thought and may play an important role in limnic and marine ecosystems. Of the different diatom-parasitic lineages, the genus Miracula is one of the most abundant in marine ecosystems. Here a species of Miracula parasitising Fragilaria capucina s.l. from Iceland is described as Miracula blauvikensis. In addition, its phylogenetic position is clarified and its life-cycle documented. The species has been brought into co-cultivation with its host, and due to the ease of cultivation and the convenient microscopy of the diatom threads, this co-culture might be a useful tool to study oomycete-diatom interactions in the future. Citation: Buaya A, Thines M (2022). Miracula blauvikensis: a new species of Miracula from Iceland, and report of a co-cultivation system for studying oomycete-diatom interactions. Fungal Systematics and Evolution 10: 169-175. doi: 10.3114/fuse.2022.10.07.

Sirolpidium bryopsidis, a parasite of green algae, is probably conspecific with Pontisma lagenidioides, a parasite of red algae.

The genus Sirolpidium (Sirolpidiaceae) of the Oomycota includes several species of holocarpic obligate aquatic parasites. These organisms are widely occurring in marine and freshwater habitats, mostly infecting filamentous green algae. Presently, all species are only known from their morphology and descriptive life cycle traits. None of the seven species classified in Sirolpidium, including the type species, S. bryopsidis, has been rediscovered and studied for their molecular phylogeny, so far. Originally, the genus was established to accommodate all parasites of filamentous marine green algae. In the past few decades, however, Sirolpidium has undergone multiple taxonomic revisions and several species parasitic in other host groups were added to the genus. While the phylogeny of the marine rhodophyte- and phaeophyte-infecting genera Pontisma and Eurychasma, respectively, has only been resolved recently, the taxonomic placement of the chlorophyte-infecting genus Sirolpidium remained unresolved. In the present study, we report the phylogenetic placement of Sirolpidium bryopsidis infecting the filamentous marine green algae Capsosiphon fulvescens sampled from Skagaströnd in Northwest Iceland. Phylogenetic reconstructions revealed that S. bryopsidis is either conspecific or at least very closely related to the type species of Pontisma, Po. lagenidioides. Consequently, the type species of genus Sirolpidium, S. bryopsidis, is reclassified to Pontisma. Further infection trials are needed to determine if Po. bryopsidis and Po. lagenidioides are conspecific or closely related. In either case, the apparently recent host jump from red to green algae is remarkable, as it opens the possibility for radiation in a largely divergent eukaryotic lineage. Citation: Buaya AT, Scholz B, Thines M (2021). Sirolpidium bryopsidis, a parasite of green algae, is probably conspecific with Pontisma lagenidioides, a parasite of red algae. Fungal Systematics and Evolution 7: 223-231. doi: 10.3114/fuse.2021.07.11.

Bolbea parasitica gen. et sp. nov., a cultivable holocarpic parasitoid of the early-diverging Saprolegniomycetes.

Holocarpic oomycetes convert their entire cytoplasm into zoospores and thus do not form dedicated sporangia or hyphal compartments for asexual reproduction. The majority of holocarpic oomycetes are obligate parasites and parasitoids of a diverse suite of organisms, among them green and red algae, brown seaweeds, diatoms, fungi, oomycetes and invertebrates. Most of them are found among the early diverging oomycetes or the Peronosporomycetes, and some in the early-diverging Saprolegniomycetes (Leptomitales). The obligate parasitism renders it difficult to study some of these organisms. Only a few members of the genus Haliphthoros s. l. have been cultured without their hosts, and of the parasitoid Leptomitales, some transient cultures have been established, which are difficult to maintain. Here, the cultivation of a new holocarpic oomycete genus of the Leptomitales, Bolbea, is presented. Bolbea is parasitic to ostracods, is readily cultivable on malt extract agar, and upon contact with water converts its cytoplasm into zoospores. Its morphology and phylogenetic relationships are reported. Due to the ease of cultivation and the ready triggering of zoospore development, similar to some lagenidiaceous oomycetes, the species could be a promising model to study sporulation processes in detail.

Two new species of the Peronospora belbahrii species complex, Pe. choii sp. nov. and Pe. salviae-pratensis sp. nov., and a new host for Pe. salviae-officinalis.

The downy mildew species parasitic to Mentheae are of particular interest, as this tribe of Lamiaceae contains a variety of important medicinal plants and culinary herbs. Over the past two decades, two pathogens, Peronospora belbahrii and Pe. salviae-officinalis have spread globally, impacting basil and common sage production, respectively. In the original circumscription of Pe. belbahrii, the downy mildew of coleus (Plectranthus scutellarioides) was ascribed to this species in the broader sense, but subtle differences in morphological and molecular phylogenetic analyses using two genes suggested that this pathogen would potentially need to be assigned to a species of its own. In the present study, Peronospora species causing downy mildew on members of the Mentheae, including clary sage (Salvia sclarea), meadow sage (S. pratensis), basil (Ocimum basilicum), ground ivy (Glechoma hederacea) and coleus (Plectranthus scutellarioides) were studied using light microscopy and molecular phylogenetic analyses based on six loci (ITS rDNA, cox1, cox2, ef1a, hsp90 and ß-tubulin) to clarify the species boundaries in the Pe. belbahrii species complex. The downy mildew on Salvia pratensis is shown to be distinct from Pe. salviae-officinalis and closely related to Pe. glechomae, and is herein described as a new species, Pe. salviae-pratensis. The downy mildew on S. sclarea was found to be caused by Pe. salviae-officinalis. This is of phytopathological importance, because meadow sage thus does not play a role as inoculum source for common sage in the natural habitat of the former in Europe and Asia, while clary sage probably does. The multi-gene phylogeny revealed that the causal agent of downy mildew on coleus is distinct from Pe. belbahrii on basil, and is herein described as a new taxon, Pe. choii.

Diatomophthoraceae - a new family of olpidiopsis-like diatom parasitoids largely unrelated to Ectrogella.

The oomycete genus Ectrogella currently comprises a rather heterogeneous group of obligate endoparasitoids, mostly of diatoms and algae. Despite their widespread occurrence, little is known regarding the phylogenetic affinities of these bizarre organisms. Traditionally, the genus was included within the Saprolegniales, based on zoospore diplanetism and a saprolegnia/achlya-like zoospore discharge. The genus has undergone multiple re-definitions in the past, and has often been used largely indiscriminately for oomycetes forming sausage-like thalli in diatoms. While the phylogenetic affinity of the polyphyletic genus Olpidiopsis has recently been partially resolved, taxonomic placement of the genus Ectrogella remained unresolved, as no sequence data were available for species of this genus. In this study, we report the phylogenetic placement of Ectrogella bacillariacearum infecting the freshwater diatom Nitzschia sigmoidea. The phylogenetic reconstruction shows that Ectrogella bacillariacearum is grouped among the early diverging lineages of the Saprolegniomycetes with high support, and is unrelated to the monophyletic diatom-infecting olpidiopsis-like species. As these species are neither related to Ectrogella, nor to the early diverging lineages of Olpidiopsis s. str. and Miracula, they are placed in a new genus, Diatomophthora, in the present study.

Miracula moenusica, a new member of the holocarpic parasitoid genus from the invasive freshwater diatom Pleurosira laevis.

Holocarpic oomycetes are poorly known but widespread parasites in freshwater and marine ecosystems. Most of the holocarpic species seem to belong to clades that diverge before the two crown lineages of the oomycetes, the Saprolegniomycetes and the Peronosporomycetes. Recently, the genus Miracula was described to accommodate Miracula helgolandica, a holocarpic parasitoid of Pseudo-nitzschia diatoms, which received varying support for its placement as the earliest-diverging oomycete lineage. In the same phylogenetic reconstruction, Miracula helgolandica was grouped with some somewhat divergent sequences derived from environmental sequencing, indicating that Miracula would not remain monotypic. Here, a second species of Miracula is reported, which was found as a parasitoid in the limnic centric diatom Pleurosira leavis. Its life-cycle stages are described and depicted in this study and its phylogenetic placement in the genus Miracula revealed. As a consequence, the newly discovered species is introduced as Miracula moenusica.

Revisiting Salisapiliaceae.

Of the diverse lineages of the Phylum Oomycota, saprotrophic oomycetes from the salt marsh and mangrove habitats are still understudied, despite their ecological importance. Salisapiliaceae, a monophyletic and monogeneric taxon of the marine and estuarine oomycetes, was introduced to accommodate species with a protruding hyaline apical plug, small hyphal diameter and lack of vesicle formation during zoospore release. At the time of description of Salisapilia, only few species of Halophytophthora, an ecologically similar, phylogenetically heterogeneous genus from which Salisapilia was segregated, were included. In this study, a revision of the genus Salisapilia is presented, and five new combinations (S. bahamensis, S. elongata, S. epistomia, S. masteri, and S. mycoparasitica) and one new species (S. coffeyi) are proposed. Further, the species description of S. nakagirii is emended for some exceptional morphological and developmental characteristics. A key to the genus Salisapilia is provided and its generic circumscription and character evolution in cultivable Peronosporales are discussed.

Holocarpic oomycete parasitoids of red algae are not Olpidiopsis.

Olpidiopsis is a genus of obligate holocarpic endobiotic oomycetes. Most of the species classified in the genus are known only from their morphology and life cycle, and a few have been examined for their ultrastructure or molecular phylogeny. However, the taxonomic placement of all sequenced species is provisional, as no sequence data are available for the type species, O. saprolegniae, to consolidate the taxonomy of species currently placed in the genus. Thus, efforts were undertaken to isolate O. saprolegniae from its type host, Saprolegnia parasitica and to infer its phylogenetic placement based on 18S rDNA sequences. As most species of Olpidiopsis for which sequence data are available are from rhodophyte hosts, we have also isolated the type species of the rhodophyte-parasitic genus Pontisma, P. lagenidioides and obtained partial 18S rDNA sequences. Phylogenetic reconstructions in the current study revealed that O. saprolegniae from Saprolegnia parasitica forms a monophyletic group with a morphologically similar isolate from S. ferax, and a morphologically and phylogenetically more divergent species from S. terrestris. However, they were widely separated from a monophyletic, yet unsupported clade containing P. lagenidioides and red algal parasites previously classified in Olpidiopsis. Consequently, all holocarpic parasites in red algae should be considered to be members of the genus Pontisma as previously suggested by some researchers. In addition, a new species of Olpidiopsis, O. parthenogenetica is introduced to accommodate the pathogen of S. terrestris.

Broad host range species in specialised pathogen groups should be treated with suspicion - a case study on Entyloma infecting Ranunculus.

Plant pathogenic smut fungi in the broader sense can be divided into the Ustilaginomycetes, which cause classical smut symptoms with masses of blackish spores being produced in a variety of angiosperms, and the Exobasidiomycetes, which are often less conspicuous, as many do not shed large amounts of blackish spores. The leaf-spot causing members of the genus Entyloma (Entylomatales, Exobasidiomycetes) belong to the latter group. Currently, 172 species that all infect eudicots are included in the genus. Vánky (2012) recognised five Entyloma species on species of Ranunculus s.lat. Two have been reported only from Ficaria verna s.lat., while three, E. microsporum, E. ranunculi-repentis, E. verruculosum, have been reported to have a broad host range, encompassing 30, 26, and 5 species of Ranunculus, respectively. This broad host range is in contrast to the generally high host specificity assumed for species of Entyloma, indicating that they may represent complexes of specialised species. The aim of this study was to investigate Entyloma on Ranunculus s.lat. using multigene phylogenies and morphological comparisons. Phylogenetic analyses on the basis of up to four loci (ITS, atp2, ssc1, and map) showed a clustering of Entyloma specimens according to host species. For some of these Entyloma lineages, names not currently in use were available and reinstated. In addition, Entyloma microsporum s.str. is neotypified. Six novel species are described in this study, namely, Entyloma jolantae on Ranunculus oreophilus, E. klenkei on R. marginatus, E. kochmanii on R. lanuginosus, E. piepenbringiae on R. polyanthemos subsp. nemorosus (type host) and R. repens, E. savchenkoi on R. paludosus, and E. thielii on R. montanus. For all species diagnostic bases and morphological characteristics are provided. The results in this study once more highlight the importance of detailed re-investigation of broad host-range pathogens of otherwise specialised plant pathogen groups.

The Genetic Structure of Pseudoperonospora cubensis Populations.

Pseudoperonospora cubensis is a destructive foliar pathogen of economically important cucurbitaceous crops in the United States and worldwide. In this study, we investigated the genetic structure of 465 P. cubensis isolates from three continents, 13 countries, 19 states of the United States, and five host species using five nuclear and two mitochondrial loci. Bayesian clustering resolved six genetic clusters and suggested some population structure by geographic origin and host, because some clusters occurred more or less frequently in particular categories. All of the genetic clusters were present in the sampling from North America and Europe. Differences in cluster occurrence were observed by country and state. Isolates from cucumber had different cluster composition and lower genetic diversity than isolates from other cucurbits. Because genetic structuring was detected, isolates that represent the genetic variation in P. cubensis should be used when developing diagnostic tools, fungicides, and resistant host varieties. Although this study provides an initial map of global population structure of P. cubensis, future genotyping of isolates could reveal population structure within specific geographic regions, across a wider range of hosts, or during different time points during the growing season.

Salisapiliaceae - a new family of oomycetes from marsh grass litter of southeastern North America.

Several filamentous oomycete species of the genus Halophytophthora have recently been described from marine environments, mostly from subtropical and tropical ecosystems. During a survey of oomycetes from leaf litter of Spartina alterniflora in salt marshes of southeastern Georgia, isolates of four taxa were recovered that bore similarity to some members of Halophytophthora but were highly divergent from isolates of Halophytophthora s.str. based on a combined sequence analysis of two nuclear loci. In phylogenetic analyses, these isolates were placed basal to a monophyletic group comprised of Pythium of the Pythiaceae and the Peronosporaceae. Sequence and morphology of these taxa diverged from the type species Halophytophthora vesicula, which was placed within the Peronosporaceae with maximum support. As a consequence a new family, the Salisapiliaceae, and a new genus, Salisapilia, are described to accommodate the newly discovered species, along with one species previously classified within Halophytophthora. Morphological features that separate these taxa from Halophytophthora are a smaller hyphal diameter, oospore production, lack of vesicle formation during sporulation, and a plug of hyaline material at the sporangial apex that is displaced during zoospore release. Our findings offer a first glance at the presumably much higher diversity of oomycetes in estuarine environments, of which ecological significance requires further exploration.

A new species of Albugo parasitic to Arabidopsis thaliana reveals new evolutionary patterns in white blister rusts (Albuginaceae).

The obligate biotrophic lineages of the white blister rusts (Albuginales, Oomycota) are of ancient origin compared to the rather recently evolved downy mildews, and sophisticated mechanisms of biotrophy and a high degree of adaptation diversity are to be expected in these organisms. Speciation in the biotrophic Oomycetes is usually thought to be the consequence of host adaptation or geographic isolation. Here we report the presence of two distinct species of Albugo on the model plant Arabidopsis thaliana, Albugo candida and Albugo laibachii, the latter being formally described in this manuscript. Both species may occupy the same host within the same environment, but are nevertheless phylogenetically distinct, as inferred from analyses of both mitochondrial and nuclear DNA sequences. Different ways of adapting to their host physiology might constitute an important factor of their different niches. Evidence for this can be gained from the completely different host range of the two pathogens. While Albugo candida is a generalist species, consisting of several physiological varieties, which is able to parasitize a great variety of Brassicaceae, Albugo laibachii has not been found on any host other than Arabidopsis thaliana. Therefore, Albugo laibachii belongs to a group of highly specialised species, like the other known specialist species in Albugo s.s., Albugo koreana, Albugo lepidii and Albugo voglmayrii. The comparative investigation of the effector genes and host targets in the generalist and the specialist species may constitute a model system for elucidating the fundamental processes involved in plant pathogen co-adaptation and speciation.

First Report of Pustula tragopogonis, the Cause of White Rust on Cultivated Sunflower in Southern Germany.

Pustula tragopogonis (Pers.) Thines (2), formerly known as Albugo tragopogonis (Pers.) Gray, causes white rust on some members of the Asteraceae. It has been reported to attack cultivated sunflower in several parts of the world, for example, North America (1) and South Africa (3), but has not been previously reported on sunflower in Germany. During June 2003, the first diseased plants were observed in a sunflower field near Stuttgart, where sunflowers were grown as ornamentals. During 2004, P. tragopogonis was found in several ornamental sunflower fields in southern Germany. In some cases, P. tragopogonis co-occurred with Plasmopara halstedii (Farlow) Berl. & De Toni on the same plants. After a cold winter, the first diseased plants for 2005 were observed during June in the area around Stuttgart. The percentage of affected plants in the sunflower fields varied considerably, ranging from 20 to 80%. Only small lesions with oospores could be found on petioles and stems of infected plants. The typical spots with white blisters were observed mostly on leaves in the upper half of the plants, but also occurred on the bracteoles of the flower heads. Spots were chlorotic on the upper side of the leaf and exhibited whitish pustules on the lower side of the leaf. Pustules were concentrated around the margins of the leaves, but also were observed scattered on the whole lower surface. Light and scanning electron microscopy, as well as polymerase chain reaction and internal transcribed spacer sequencing were applied to identify the attacking pathogen as P. tragopogonis. Infection tests under laboratory conditions also proved the pathogenicity of the attacking agent to oilseed sunflower. Since the pathogen has survived the harsh winter months of early 2005 and appeared in three consecutive seasons, the disease seems now to be established in southern Germany and is expected to spread into other regions. References: (1) T. J. Gulya et al. Plant Dis. 86:559, 2002. (2) M. Thines and O. Spring. Mycotaxon 92:443, 2005. (3) P. S. van Wyk et al. Helia 22:83, 1995.