ANTHER SMUT DISEASE IN DIANTHUS SILVESTER (CARYOPHYLLACEAE): NATURAL SELECTION ON FLORAL TRAITS

Mating opportunities, pollination intensity, and pollen dispersal ability may vary with variation in floral traits such as color, size, and shape. Where these traits are selected by pollinators for enhanced elaboration, they should evolve toward the equilibrium between selection for further elaboration and selection against this through reduced fecundity or vitality. Here we show that pollinator-borne fungal diseases of plants may be a factor influencing the position of this equilibrium. Populations of the rock pink, Dianthus silvester often contain individuals infected with the anther smut fungus Microbotryum violaceum (= Ustilago violacea). In a naturally infected population in the Alps of eastern Switzerland we investigated how intrapopulation variation in flower size and nectar rewards influenced spore deposition and how floral traits varied with disease status. We found that spore deposition increased with increasing petal size, suggesting that large-flowered plants were at a greater risk of disease. Spore deposition was also higher for plants growing in patches with many or a high proportion of diseased neighbors. Multiple regression analyses showed that petal size or nectar reward influenced spore deposition when the effects of neighborhood disease abundance were controlled statistically. In sequential analyses, after removing the effects of disease density or frequency and plant gender, petal length explained significant variation in spore deposition. Diseased plants had reduced female reproductive organs, but calyx size was intermediate between that of healthy perfect and female flowers of this gynodioecious-gynomonoecious species, and diseased plants bore flowers with the largest petals. This may reflect a symptom of this disease or the cause, if larger-flowered plants are more likely to become infected. We conclude that investment to pollinator attraction may bring an enhanced risk of contracting this sterilizing pollinator-borne disease, so natural selection by the fungus M. violaceum acts to lower attractiveness to pollinators.     

Distribution and population structure of the anther smut Microbotryum silenes‐acaulis parasitizing an arctic–alpine plant

Cold‐adapted organisms with current arctic–alpine distributions have persisted during the last glaciation in multiple ice‐free refugia, leaving footprints in their population structure that contrast with temperate plants and animals. However, pathogens that live within hosts having arctic–alpine distributions have been little studied. Here, we therefore investigated the geographical range and population structure of a fungus parasitizing an arctic–alpine plant. A total of 1437 herbarium specimens of the plant Silene acaulis were examined, and the anther smut pathogen Microbotryum silenes‐acaulis was present throughout the host's geographical range. There was significantly greater incidence of anther smut disease in more northern latitudes and where the host locations were less dense, indicating a major influence of environmental factors and/or host demographic structure on the pathogen distribution. Genetic analyses with seven microsatellite markers on recent collections of 195 M. silenes‐acaulis individuals revealed three main genetic clusters, in North America, northern Europe and southern Europe, likely corresponding to differentiation in distinct refugia during the last glaciation. The lower genetic diversity in northern Europe indicates postglacial recolonization northwards from southern refugia. This study combining herbarium surveys and population genetics thus uniquely reveals the effects of climate and environmental factors on a plant pathogen species with an arctic–alpine distribution.     

Lower prevalence but similar fitness in a parasitic fungus at higher radiation levels near Chernobyl

Nuclear disasters at Chernobyl and Fukushima provide examples of effects of acute ionizing radiation on mutations that can affect the fitness and distribution of species. Here, we investigated the prevalence of Microbotryum lychnidis‐dioicae, a pollinator‐transmitted fungal pathogen of plants causing anther‐smut disease in Chernobyl, its viability, fertility and karyotype variation, and the accumulation of nonsynonymous mutations in its genome. We collected diseased flowers of Silene latifolia from locations ranging by more than two orders of magnitude in background radiation, from 0.05 to 21.03 μGy/h. Disease prevalence decreased significantly with increasing radiation level, possibly due to lower pollinator abundance and altered pollinator behaviour. Viability and fertility, measured as the budding rate of haploid sporidia following meiosis from the diploid teliospores, did not vary with increasing radiation levels and neither did karyotype overall structure and level of chromosomal size heterozygosity. We sequenced the genomes of twelve samples from Chernobyl and of four samples collected from uncontaminated areas and analysed alignments of 6068 predicted genes, corresponding to 1.04 × 10⁷ base pairs. We found no dose‐dependent differences in substitution rates (neither dN, dS, nor dN/dS). Thus, we found no significant evidence of increased deleterious mutation rates at higher levels of background radiation in this plant pathogen. We even found lower levels of nonsynonymous substitution rates in contaminated areas compared to control regions, suggesting that purifying selection was stronger in contaminated than uncontaminated areas. We briefly discuss the possibilities for a mechanistic basis of radio resistance in this nonmelanized fungus.     

Ein Beitrag zur Systematik und Entwicklungsbiologie höherer Pilze: Hefe-Typen der Basidiomyceten. Teil II: Microbotryum-Typ

In this and three further papers 205 yeasts and yeast states of Basidiomycetes and presumed relatives were investigated comparatively on the basis of the carbohydrate (neutral sugars) pattern of purified cell walls, urease-activity, diazonium blue B reaction on the production of extracellular amyloid compounds (EAS), fermentation of carbohydrates, and ubiquinone data. A clustering leading to the Protomyces-, the Microbotryum-, the Ustilago-, the Dacrymyces-, and the Tremella-type became apparent, especially from the qualitative and quantitative cell wall carbohydrate pattern. The different yeast types correspond well with 5S rRNA clusters known from the literature. 31 strains clustering within the Microbotryum-type comprise the phragmobasidial smut fungi of dicotyledonous hosts (Microbotryum. Sphacelotheca), the phragmobasidial Rhodosporidium- and Leucosporidium-species including some anamorph Rhodotorula-species, which lack an oxidative degradation of myo-inositol, the genera Sporobolomyces and Sporidiobolus, the Septobasidiales and some simple septate Auriculariales e.g. Agaricostilbum, Platygloea. Main characteristics of the Microbotryum-type are: 1. The absence of extracellular amyloid compounds. 2. The dominance of mannose and the presence of fucose as cell wall constituents. 3. A positive DBB-reaction and splitting of urea. Four Ustilago species parasitic on dicotyledonous hosts were transfered to Microbotryum (M. scabiosae, M. scorzonerae, M. cordae, M. vinosum) as a consequence from cell wall carbohydrate composition, production of rhodotorulic acid, and 5S rRNA sequence data from the literature. The predominance of mannose in the cell wall — otherwise only known from ascomycetous yeasts –, a type A secondary structure of 5S rRNA, a simple unifactorial mating system in all parasitic smut species suggest that the Microbotryum-type might be ancestral to the Ustilago-type. An evolution of simple (“siphonal”) holobasidia from “pseudotrichal” phragmobasidia will be discussed.     

The tempo and modes of evolution of reproductive isolation in fungi

Reproductive isolation is an essential ingredient of speciation, and much has been learned in recent years about the evolution of reproductive isolation and the genetics of reproductive barriers in animals and plants. Fungi have been neglected on these aspects, despite being tractable model eukaryotes. Here, we used a model fitting approach to look at the importance of different barriers to gene flow to explain the decrease of reproductive compatibility with genetic distance in fungi. We found support for the occurrence of reinforcement in the presyngamy compatibility among basidiomycetes. In contrast, no evidence for reinforcement was detected in ascomycetes, concurring with the idea that host/habitat adaptation in this group can pleiotropically cause reproductive isolation. We found no evidence of a snowballing accumulation of postsyngamic reproductive incompatibilities in either ascomycetes or the complex of anther smut fungi. Together with previous studies, our results suggest that ecologically based barriers to gene flow and karyotypic differences may have an important role in hybrid inviability and sterility in fungi. Interestingly, hybrid sterility appeared to evolve faster than hybrid inviability in fungi.     

Seed fitness of hermaphrodites in areas with females and anther smut disease: Silene acaulis and Microbotryum violaceum

Sex-dependent infection rates could change the effective sex ratio of a population. Here, I tested whether females and hermaphrodites of Silene acaulis were equally likely to be infected by Microbotryum violaceum, a fungus that sterilizes the host, and whether sex allocation in hermaphrodites differed between low and high disease plots. Sex ratios of healthy and diseased plants were estimated in five natural plots. Fitness gained through seed production was estimated by measuring seed quantity and quality for each sex morph in eight plots for 2 yr; four plots had 1-5% disease frequency and four plots had 18-25% disease frequency. Sex ratios of healthy and diseased plants did not differ in five plots. The proportion of fitness hermaphrodites gained through ovules varied from 25 to 48%, indicating that this population is near the cosexual end of gynodioecy. Variation in functional gender of hermaphrodites was not explained by sex-dependent infection rates. Spatial heterogeneity in resources and microclimate seems to be important in explaining both disease frequency and variation in seed production by females and hermaphrodites.     

Natural selection on floral traits of female Silene dioica by a sexually transmitted disease

Floral traits endowing high reproductive fitness can also affect the probability of plants contracting sexually transmitted diseases. We explore how variations in floral traits influence the fitness of Silene dioica females in their interactions with pollinators carrying pollen or spores of the sterilizing anther-smut fungus Microbotryum violaceum. We collected healthy and infected plants in a highly diseased population and grew them under conditions that 'cure' infected individuals, and used standard regression methods to detect natural selection on floral traits. Narrow-sense heritabilities, coefficients of additive genetic variation (CV(A)) and genetic correlations among traits were estimated from paternal half-sib groups. Pollinator preferences imposed strong direct and directional selection on traits affecting female attractiveness and pollen-/spore-capturing abilities. Levels of additive genetic variance were high in these traits, suggesting that rapid responses to selection are possible. By considering our results in the light of spatial and temporal heterogeneity resulting from the colonization dynamics typical for this species, we suggest that the conflicting selective effects of pollen/spore loads lead to the maintenance of genetic variation in these traits.     

Within‐ and among‐population variation in infectivity,latency and spore production in a host–pathogen system

In spatially structured populations, host–parasite coevolutionary potential depends on the distribution of genetic variation within and among populations. Inoculation experiments using the plant, Silene latifolia, and its fungal pathogen, Microbotryum violaceum, revealed little overall differentiation in infectivity/resistance, latency or spore production among host or pathogen populations. Within populations, fungal strains had similar means, but varied in performance across plant populations. Variation in resistance among seed families indicates the potential for parasite‐mediated selection, whereas there was little evidence for local pathogen genotype × plant genotype interactions assumed by most theoretical coevolution models. Lower spore production on sympatric than allopatric hosts confirmed local fungal maladaptation already observed for infectivity. Correlations between infectivity and latency or spore production suggest a common mechanism for variation in these traits. Our results suggest low variation available to this pathogen for tracking its coevolving host. This may be caused by random drift, breeding system or migration characteristic of metapopulation dynamics.     

Spore Liberation and Dispersal in Smut Fungi*

Teliospores are the most important diaspores of smut fungi, albeit not the only ones. The role of basidla, basidiospores, secondary spores, yeast cells, and infected parts of the host for dispersal has often been neglected. Many smut species have soral structures like galls, peridia, and elaters, which cause teliospores to be liberated over prolonged periods. This increases the chance that at least some spores are released under favourable wet climatic conditions and while host plants are susceptible. In this review, the diversity of dispersal units as well as vectors of smut fungi are presented. The importance of timing of diaspore liberation, flexibility in dispersal strategies, and the genetic and evolutionary implications of dispersal strategies of smut fungi are discussed. The general considerations are complemented by examples based on original field and laboratory observations: peridia of Farysia corniculata and certain species of Sporisorium expose the spore mass by hygroscopic movement under wet conditions (hygrochasy) favourable for teliospore germination and infection of a host plant. Basidia with firmly attached basidiospores liberated from spore balls of Doassansiopsis deformans, branched basidiospores of Rhamphospora nymphaeae, needle-shaped basidiospores of species of Entyloma, folded basidiospores of Mycosyrinx cissi, and stellate groups of yeast cells of Trichocintractia utriculicola show enlarged surfaces, which are advantageous for dispersal in water. Galls filled with spore balls of Doassansiopsis limnocharidis and witches' brooms formed by spikelets infected by Cintractia standleyana separate from the host and fall into water where they are dispersed.