Sensitivity of spiders from different ecosystems to lambda-cyhalothrin: effects of phylogeny and climate.

BACKGROUND: In spite of their importance as arthropod predators, spiders have received little attention in the risk assessment of pesticides. In addition, research has mainly focused on a few species commonly found in agricultural habitats. Spiders living in more natural ecosystems may also be exposed to and affected by pesticides, including insecticides. However, their sensitivity and factors driving possible variations in sensitivity between spider taxa are largely unknown. To fill this gap, we quantified the sensitivity of 28 spider species from a wide range of European ecosystems to lambda-cyhalothrin in an acute exposure scenario. RESULTS: Sensitivity varied among the tested populations by a factor of 30. Strong differences in sensitivity were observed between families, but also between genera within the Lycosidae. Apart from the variation explained by the phylogeny, spiders from boreal and polar climates were more sensitive than spiders from warmer areas. Overall, the median lethal concentration (LC50 ) of 85% of species was below the recommended application rate of lambda-cyhalothrin (75 ng a.i. cm-2 ). CONCLUSION: Our study underlines the high sensitivity of spiders to lambda-cyhalothrin, which can lead to unintended negative effects on pest suppression in areas treated with this insecticide. The strong differences observed between families and genera indicate that the functional composition of spider communities would change in affected areas. Overall, the variation in spider sensitivity suggests that multispecies investigations should be more widely considered in pesticide risk assessment. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Copper Uptake and Its Effects on Two Riparian Plant Species, the Native Urtica dioica, and the Invasive Fallopia japonica.

Copper accumulating in stream sediments can be transported to adjacent riparian habitats by flooding. Although being an essential element for plants, copper is toxic at high concentrations and restricts, among other things, plant growth. Besides copper, invasive plants, such as Fallopia japonica, which are known to be tolerant toward heavy metals, modify riparian habitats. If the tolerance of F. japonica is higher compared to native plants, this could accelerate invasion under high heavy metal stress. Therefore, we aimed to compare the effect of copper on two common riparian plants, the invasive F. japonica and the native Urtica dioica. We performed a pot experiment with a gradient from 0 to 2430 mg kg-1 of soil copper. We hypothesized that (i) negative effects on plant growth increase with increasing soil copper concentrations with F. japonica being less affected and (ii) accumulating higher amounts of copper in plant tissues compared to U. dioica. In support of our first hypothesis, growth (height, leaf number) and biomass (above- and belowground) of F. japonica were impacted at the 810 mg kg-1 treatment, while the growth of U. dioica was already impacted at 270 mg kg-1. Due to 100% mortality of plants, the 2430 mg kg-1 treatment was omitted from the analysis. In contrast, chlorophyll content slightly increased with increasing copper treatment for both species. While U. dioica accumulated more copper in total, the copper uptake by F. japonica increased more strongly after exposure compared to the control. In the 810 mg kg-1 treatment, copper concentrations in F. japonica were up to 2238% higher than in the control but only up to 634% higher in U. dioica. Our results indicate that F. japonica might be able to more efficiently detoxify internal copper concentrations controlling heavy metal effects compared to the native species. This could give F. japonica a competitive advantage particularly in polluted areas, facilitating its invasion success.

Phylogenetic relationships in the genus Podoserpula and description of P. aliweni, a new species from Chile.

Phylogenetic analyses based on a three-locus nuclear data set (ITS1-5.8S-ITS2, the 5' end of the 28S, and the largest subunit of RNA polymerase I) supported the pagoda fungus (Podoserpula, Amylocorticiales) as a monophyletic group most closely related to species of Anomoporia, which is nonmonophyletic, and Amyloathelia. Phylogenetic relationships inferred from internal transcribed spacer (ITS) sequences of specimens sampled in Australia, Chile, China, Madagascar, and New Zealand divided Podoserpula into two major lineages: Clade A containing Australian and New Zealand collections designated P. pusio and the Chinese species P. ailaoshanensis, which have basidiospores with no reaction to Melzer's reagent, and Clade B, which includes a species described from Chile, P. aliweni, and specimens originating from Australia, Chile, Madagascar, and New Zealand with dextrinoid basidiospores. Podoserpula aliweni forms a unique branch in the phylogenetic tree and differs from its most closely related taxon by 1.8-2.1% in the ITS region. The new species exhibits a tree-like habit with a white to concolorous stipe-like base with the hymenophore's main subcylindrical axis bearing up to 18 superimposed pilei, slightly enrolled white margins shading from yellowish white to orange-yellow toward the center, and ellipsoid to broadly ellipsoid basidiospores measuring 4.0-4.5 × 3.5 µm. Ecologically, P. aliweni occurs during the rainy season, often gregariously on dried branches or wet soil under Nothofagus dombeyi or N. obliqua and has a distribution range of more than 600 km in southern Chile. This study extends the known distributional range and increases our knowledge on the phylogenetic diversity and taxonomy in Podoserpula.

Determining threshold values for barcoding fungi: lessons from Cortinarius (Basidiomycota), a highly diverse and widespread ectomycorrhizal genus.

Different distance-based threshold selection approaches were used to assess and compare use of the internal transcribed spacer (ITS) region to distinguish among 901 Cortinarius species represented by >3000 collections. Sources of error associated with genetic markers and selection approaches were explored and evaluated using MOTUs from genus and lineage based-alignments. Our study indicates that 1%-2% more species can be distinguished by using the full-length ITS barcode as compared to either the ITS1 or ITS2 regions alone. Optimal threshold values for different picking approaches and genetic marker lengths inferred from a subset of species containing major lineages ranged from 97.0% to 99.5% sequence similarity using clustering optimization and UNITE SH, and from 1% to 2% sequence dissimilarity with CROP. Errors for the optimal cutoff ranged from 0% to 70%, and these can be reduced to a maximum of 22% when excluding species lacking a barcode gap. A threshold value of 99% is suitable for distinguishing species in the majority of lineages in the genus using the entire ITS region but only 90% of the species could be identified using just the ITS1 or ITS2 region. Prior identification of species, lacking barcode gaps and their subsequent separate analyses, maximized the accuracy of threshold approaches.

Divergence Times and Phylogenetic Patterns of Sebacinales, a Highly Diverse and Widespread Fungal Lineage.

Patterns of geographic distribution and composition of fungal communities are still poorly understood. Widespread occurrence in terrestrial ecosystems and the unique richness of interactions of Sebacinales with plants make them a target group to study evolutionary events in the light of nutritional lifestyle. We inferred diversity patterns, phylogenetic structures and divergence times of Sebacinales with respect to their nutritional lifestyles by integrating data from fossil-calibrated phylogenetic analyses. Relaxed molecular clock analyses indicated that Sebacinales originated late Permian within Basidiomycota, and their split into Sebacinaceae and Serendipitaceae nom. prov. likely occurred during the late Jurassic and the early Cretaceous, coinciding with major diversifications of land plants. In Sebacinaceae, diversification of species with ectomycorrhizal lifestyle presumably started during the Paleocene. Lineage radiations of the core group of ericoid and cavendishioid mycorrhizal Sebacinales started probably in the Eocene, coinciding with diversification events of their hosts. The diversification of Sebacinales with jungermannioid interactions started during the Oligocene, and occurred much later than the diversification of their hosts. Sebacinales communities associated either with ectomycorrhizal plants, achlorophyllous orchids, ericoid and cavendishioid Ericaceae or liverworts were phylogenetically clustered and globally distributed. Major Sebacinales lineage diversifications started after the continents had drifted apart. We also briefly discuss dispersal patterns of extant Sebacinales.

On the Evolutionary History of Uleiella chilensis, a Smut Fungus Parasite of Araucaria araucana in South America: Uleiellales ord. nov. in Ustilaginomycetes.

The evolutionary history, divergence times and phylogenetic relationships of Uleiella chilensis (Ustilaginomycotina, smut fungi) associated with Araucaria araucana were analysed. DNA sequences from multiple gene regions and morphology were analysed and compared to other members of the Basidiomycota to determine the phylogenetic placement of smut fungi on gymnosperms. Divergence time estimates indicate that the majority of smut fungal orders diversified during the Triassic-Jurassic period. However, the origin and relationships of several orders remain uncertain. The most recent common ancestor between Uleiella chilensis and Violaceomyces palustris has been dated to the Lower Cretaceous. Comparisons of divergence time estimates between smut fungi and host plants lead to the hypothesis that the early Ustilaginomycotina had a saprobic lifestyle. As there are only two extant species of Araucaria in South America, each hosting a unique Uleiella species, we suggest that either coevolution or a host shift followed by allopatric speciation are the most likely explanations for the current geographic restriction of Uleiella and its low diversity. Phylogenetic and age estimation analyses, ecology, the unusual life-cycle and the peculiar combination of septal and haustorial characteristics support Uleiella chilensis as a distinct lineage among the Ustilaginomycotina. Here, we describe a new ustilaginomycetous order, the Uleiellales to accommodate Uleiella. Within the Ustilaginomycetes, Uleiellales are sister taxon to the Violaceomycetales.

Comparative Genomics Including the Early-Diverging Smut Fungus Ceraceosorus bombacis Reveals Signatures of Parallel Evolution within Plant and Animal Pathogens of Fungi and Oomycetes.

Ceraceosorus bombacis is an early-diverging lineage of smut fungi and a pathogen of cotton trees (Bombax ceiba). To study the evolutionary genomics of smut fungi in comparison with other fungal and oomycete pathogens, the genome of C. bombacis was sequenced and comparative genomic analyses were performed. The genome of 26.09 Mb encodes for 8,024 proteins, of which 576 are putative-secreted effector proteins (PSEPs). Orthology analysis revealed 30 ortholog PSEPs among six Ustilaginomycotina genomes, the largest groups of which are lytic enzymes, such as aspartic peptidase and glycoside hydrolase. Positive selection analyses revealed the highest percentage of positively selected PSEPs in C. bombacis compared with other Ustilaginomycotina genomes. Metabolic pathway analyses revealed the absence of genes encoding for nitrite and nitrate reductase in the genome of the human skin pathogen Malassezia globosa, but these enzymes are present in the sequenced plant pathogens in smut fungi. Interestingly, these genes are also absent in cultivable oomycete animal pathogens, while nitrate reductase has been lost in cultivable oomycete plant pathogens. Similar patterns were also observed for obligate biotrophic and hemi-biotrophic fungal and oomycete pathogens. Furthermore, it was found that both fungal and oomycete animal pathogen genomes are lacking cutinases and pectinesterases. Overall, these findings highlight the parallel evolution of certain genomic traits, revealing potential common evolutionary trajectories among fungal and oomycete pathogens, shaping the pathogen genomes according to their lifestyle.

Entorrhizomycota: A New Fungal Phylum Reveals New Perspectives on the Evolution of Fungi.

Entorrhiza is a small fungal genus comprising 14 species that all cause galls on roots of Cyperaceae and Juncaceae. Although this genus was established 130 years ago, crucial questions on the phylogenetic relationships and biology of this enigmatic taxon are still unanswered. In order to infer a robust hypothesis about the phylogenetic position of Entorrhiza and to evaluate evolutionary trends, multiple gene sequences and morphological characteristics of Entorrhiza were analyzed and compared with respective findings in Fungi. In our comprehensive five-gene analyses Entorrhiza appeared as a highly supported monophyletic lineage representing the sister group to the rest of the Dikarya, a phylogenetic placement that received but moderate maximum likelihood and maximum parsimony bootstrap support. An alternative maximum likelihood tree with the constraint that Entorrhiza forms a monophyletic group with Basidiomycota could not be rejected. According to the first phylogenetic hypothesis, the teliospore tetrads of Entorrhiza represent the prototype of the dikaryan meiosporangium. The alternative hypothesis is supported by similarities in septal pore structure, cell wall and spindle pole bodies. Based on the isolated phylogenetic position of Entorrhiza and its peculiar combination of features related to ultrastructure and reproduction mode, we propose a new phylum Entorrhizomycota, for the genus Entorrhiza, which represents an apparently widespread group of inconspicuous fungi.

Identification of a new order of root-colonising fungi in the Entorrhizomycota: Talbotiomycetales ord. nov. on eudicotyledons.

The recently described fungal phylum Entorrhizomycota was established solely for the genus Entorrhiza, species of which cause root-galls in Cyperaceae and Juncaceae. Talbotiomyces calosporus (incertae sedis) shares morphological characteristics and an ecological niche with species of Entorrhiza. We investigated the higher classification of T. calosporus to determine whether it belongs in Entorrhizomycota. Ribosomal DNA sequences showed Talbotiomyces to be a close relative of Entorrhiza and both taxa form a highly supported monophyletic group. Based on molecular phylogenetic analyses and in congruence with existing morphological and ecological data, Entorrhiza and Talbotiomyces represent a deep dichotomy within the Entorrhizomycota. While species of Entorrhiza are characterised by dolipores and occur on monocotyledons, members of Talbotiomyces are characterised by simple pores and are associated with eudicotyledons. This expands the host range of the recently described Entorrhizomycota from Poales to other angiosperms. Higher taxa, namely Talbotiomycetales ord. nov. and Talbotiomycetaceae fam. nov., are proposed here to accommodate Talbotiomyces.

Communities of endophytic sebacinales associated with roots of herbaceous plants in agricultural and grassland ecosystems are dominated by Serendipita herbamans sp. nov.

Endophytic fungi are known to be commonly associated with herbaceous plants, however, there are few studies focusing on their occurrence and distribution in plant roots from ecosystems with different land uses. To explore the phylogenetic diversity and community structure of Sebacinales endophytes from agricultural and grassland habitats under different land uses, we analysed the roots of herbaceous plants using strain isolation, polymerase chain reaction (PCR), transmission electron microscopy (TEM) and co-cultivation experiments. A new sebacinoid strain named Serendipita herbamans belonging to Sebacinales group B was isolated from the roots of Bistorta vivipara, which is characterized by colourless monilioid cells (chlamydospores) that become yellow with age. This species was very common and widely distributed in association with a broad spectrum of herbaceous plant families in diverse habitats, independent of land use type. Ultrastructurally, the presence of S. herbamans was detected in the cortical cells of Plantago media, Potentilla anserina and Triticum aestivum. In addition, 13 few frequent molecular operational taxonomic units (MOTUs) or species were found across agricultural and grassland habitats, which did not exhibit a distinctive phylogenetic structure. Laboratory-based assays indicate that S. herbamans has the ability to colonize fine roots and stimulate plant growth. Although endophytic Sebacinales are widely distributed across agricultural and grassland habitats, TEM and nested PCR analyses reinforce the observation that these microorganisms are present in low quantity in plant roots, with no evidence of host specificity.

High genetic diversity at the regional scale and possible speciation in Sebacina epigaea and S. incrustans.

BACKGROUND: Phylogenetic studies, particularly those based on rDNA sequences from plant roots and basidiomata, have revealed a strikingly high genetic diversity in the Sebacinales. However, the factors determining this genetic diversity at higher and lower taxonomic levels within this order are still unknown. In this study, we analysed patterns of genetic variation within two morphological species, Sebacina epigaea and S. incrustans, based on 340 DNA haplotype sequences of independent genetic markers from the nuclear (ITS + 5.8S + D1/D2, RPB2) and mitochondrial (ATP6) genomes for 98 population samples. By characterising the genetic population structure within these species, we provide insights into species boundaries and the possible factors responsible for genetic diversity at a regional geographic scale. RESULTS: We found that recombination events are relatively common between natural populations within Sebacina epigaea and S. incrustans, and play a significant role in generating intraspecific genetic diversity. Furthermore, we also found that RPB2 and ATP6 genes display higher levels of intraspecific synonymous polymorphism. Phylogenetic and demographic analyses based on nuclear and mitochondrial loci revealed three distinct phylogenetic lineages within of each of the morphospecies S. epigaea and S. incrustans: one major and widely distributed lineage, and two geographically restricted lineages, respectively. We found almost no differential morphological or ecological characteristics that could be used to discriminate between these lineages. CONCLUSIONS: Our results suggest that recombination and negative selection have played significant roles in generating genetic diversity within these morphological species at small geographical scales. Concordance between gene genealogies identified lineages/cryptic species that have evolved independently for a relatively long period of time. These putative species were not associated with geographic provenance, geographic barrier, host preference or distinct phenotypic innovations.

Phylogenetic diversity and structure of sebacinoid fungi associated with plant communities along an altitudinal gradient.

To study the diversity and phylogenetic structure of Sebacinales communities from eight vegetation communities along an altitudinal gradient in the Bavarian Alps (Germany), we analysed 456 thalli or roots of plants. We detected 264 sebacinoid sequences, spanning the intergenic transcribed spacer region, 5.8S and D1/D2 regions of the nuclear rRNA gene, mostly using a nested PCR approach. Based on 97% sequence similarity, 73 Sebacinales molecular taxonomic units were found from 70 host species belonging to 44 plant families. Twenty-six molecular taxonomic units represented singletons, the most frequent of these being restricted exclusively to either wooded or grassland habitats. Although Sebacinales appear to occur in low abundance in plant roots, these microorganisms are phylogenetically diverse and widely spread in the ecosystems studied. Ordination analyses showed that land use, pH and humus content strongly influence the diversity and assembly of Sebacinales. In most cases, Sebacinales communities in ecosystems with extreme soil conditions or intensive land use exhibited significant phylogenetic clustering, whereas in undisturbed plant communities no trend was observed. These results suggest that ecosystem disturbance and environmental forces have an influence on the diversity and structure of Sebacinales community assembly over local and spatial scales.

Sebacinales everywhere: previously overlooked ubiquitous fungal endophytes.

Inconspicuous basidiomycetes from the order Sebacinales are known to be involved in a puzzling variety of mutualistic plant-fungal symbioses (mycorrhizae), which presumably involve transport of mineral nutrients. Recently a few members of this fungal order not fitting this definition and commonly referred to as 'endophytes' have raised considerable interest by their ability to enhance plant growth and to increase resistance of their host plants against abiotic stress factors and fungal pathogens. Using DNA-based detection and electron microscopy, we show that Sebacinales are not only extremely versatile in their mycorrhizal associations, but are also almost universally present as symptomless endophytes. They occurred in field specimens of bryophytes, pteridophytes and all families of herbaceous angiosperms we investigated, including liverworts, wheat, maize, and the non-mycorrhizal model plant Arabidopsis thaliana. They were present in all habitats we studied on four continents. We even detected these fungi in herbarium specimens originating from pioneering field trips to North Africa in the 1830s/40s. No geographical or host patterns were detected. Our data suggest that the multitude of mycorrhizal interactions in Sebacinales may have arisen from an ancestral endophytic habit by specialization. Considering their proven beneficial influence on plant growth and their ubiquity, endophytic Sebacinales may be a previously unrecognized universal hidden force in plant ecosystems.