Species Complex Research Articles

Systematics and distribution of the genus Darevskia Arribas, 1997 on the Iranian plateau: a review

Two hundred ninety species of lacertids are presently placed in from 24 to about 30 genera. Most of the genera that have been recognized appear to be clades on the basis of their morphology, with the exception of the paraphyletic central African Adolfus and Lacerta sensu lato of Europe, Asia and northwest Africa. Traditionally, species boundaries are only based on morphological and appearance differences, and this approach is qualitatively or quantitatively investigated. At the same time, several new and modern methods can be used for a more realistic and logical investigation in the determination of species boundaries. Molecular analysis has provided more evidence about lacertids' kinship relationships. These methods work particularly well when dealing with cryptic species or species complexes. Where there is a high general and morphological similarity between the species of a population. The rock lizards, genus Darevskia Arribas, 1999, are such species, as many of its members have few distinctive and diagnostic morphological features. Biodiversity and species diversity assessment allows for more precise determination of the causes of many biological processes. These processes involve complex patterns that are especially observed in areas with high species diversity. Anatolia and Caucasus regions are two important geographical regions where Darevskia species live, which have complex morphology and unique speciation. Darevskia valentini and D. rudis are two widely distributed species of this genus, both of which have controversial taxonomy. In this study, an attempt has been made to summarize the species available in Iran.

Morphological and multi-gene phylogenetic analyses of two new species of Phyllosticta (Phyllostictaceae, Botryosphaeriales) from Southwestern China

Phyllosticta is an important genus of pathogenic fungi with worldwide distribution. Phyllosticta species also occur as endophytes and saprobes on various plant hosts. Multi-gene phylogenetic analysis based on sequence data of ITS, LSU, TEF1 and morphological evidence supported the introduction of two new species, which were isolated from dead leaves in China. Phylogenetically, the new species, P. savannaensis and P. ovalina, formed two well-separated lineages in the Phyllosticta capitalensis and Phyllosticta cruenta species complexes, respectively. P. savannaensis is closely related to P. capitalensis, but formed a separate branch (BS= 100%/BYPP= 1.0), while P. ovalina and P. hubeiensis clustered together with high support (BS= 100%/BYPP= 1.0). Both new species exhibited the typical characteristics of the genus Phyllosticta but could be distinguished from closely related species. P. savannaensis can be differentiated from the sexual stage of other Phyllosticta species by the length to width ratio of the ascospores and shape of appendages. The ascospores of P. ovalina are oval, which distinguish it from other species of Phyllosticta.

Role of biofilms in antimicrobial resistance of the bacterial bovine respiratory disease complex.

An increase in chronic, non-responsive bovine respiratory disease (BRD) infections in North American feedlot cattle is observed each fall, a time when cattle are administered multiple antimicrobial treatments for BRD. A number of factors are responsible for BRD antimicrobial treatment failure, with formation of biofilms possibly being one. It is widely accepted that biofilms play a role in chronic infections in humans and it has been hypothesized that they are the default lifestyle of most bacteria. However, research on bacterial biofilms associated with livestock is scarce and significant knowledge gaps exist in our understanding of their role in AMR of the bacterial BRD complex. The four main bacterial species of the BRD complex, Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis are able to form biofilms in vitro and there is evidence that at least H. somni retains this ability in vivo. However, there is a need to elucidate whether their biofilm-forming ability contributes to pathogenicity and antimicrobial treatment failure of BRD. Overall, a better understanding of the possible role of BRD bacterial biofilms in clinical disease and AMR could assist in the prevention and management of respiratory infections in feedlot cattle. We review and discuss the current knowledge of BRD bacteria biofilm biology, study methodologies, and their possible relationship to AMR.

Review of the subgenus Lyrothorax of the genus Pterostichus (Coleoptera: Carabidae) in the Western Palaearctic, with descriptions of two new species

Until now, the subgenus Lyrothorax Chaudoir, 1838 (genus Pterostichus Bonelli, 1810) in the Western Palaearctic included only the type species, P. caspius (Ménétriés, 1832), distributed along the southern Caspian Sea in Azerbaijan and Iran and, as an introduced species, in the Czech Republic. Studying rich material, including the types, it was found that under the name P. caspius there is a species complex comprising three closely related species, two of these are described in this paper: P. fritzei sp. n. from Iran (type locality Naharkhoran, Gorgan) and P. vagus sp. n. from Iran, which was also introduced to the Czech Republic (type locality Shaskola, Gorgan). These new species differ clearly from each other, as well as from P. caspius, mainly in the features of the median lobe of the aedeagus and its endophallus. A lectotype of P. caspius is designated to ensure the stability of the nomenclature. An identification key, images of all three species and distribution maps are provided. The occurrence of species of Lyrothorax in the Iranian province of North Khorasan and Turkmenistan is not obvious and requires confirmation.

Draft genome sequences of 148 Klebsiella pneumoniae species complex members from bovine and human hosts.

Klebsiella pneumoniae species complex members, particularly K. pnemoniae sensu stricto, are common bovine clinical mastitis pathogens and often the cause of hospital- and community-acquired infections in humans. Here, we present 148 draft genome assemblies and annotations of K. pneumoniae species complex members from bovine and human hosts in Canada.

Re-appearance of the gregarious gastropod Dendropoma anguliferum (Vermetidae) on abrasion platforms in the Carmel coast (Northern Israel)

After three decades of sharp decline in the Dendropoma populations in Israel and genuine concern for their extinction, we affirm the re-appearance of the gregarious gastropod Dendropoma anguliferum on abrasion platforms in the Carmel coast (Northern Israel). A visual survey conducted during December 2020 and February 2021 along the rims of abrasion platforms in the Carmel coast area revealed 195 living clusters of live D. anguliferum. DNA sequences from tissue samples unambiguously supported the identity of our material as the Levantine Sea member D. anguliferum of the Dendropoma petraeum species complex, with 99- 100% identity to sequences obtained from D. anguliferum from Southern Lebanon.

First report of Fusarium falciforme causing root rot and wilt on strawberry in Sinaloa, Mexico.

In Mexico strawberry production has great economic importance for the local and export markets as the country is the main strawberry supplier to the United States (SIAP, 2020). In 2022, strawberry plants with yellowing and wilting leaves, root rot and wilting, necrosis of vascular bundles and small fruits symptoms were observed in different commercial fields in the north-central Mexican state of Sinaloa, causing yield losses of about 10%. Typical Fusarium spp. colonies were recovered from all samples. They produced abundant white aerial mycelium with cream to orange pigment and branched septate hyphae (Fig. 1) (Leslie and Summerell, 2006). A total of 18 monosporic isolates were obtained by serial dilutions. The 18 isolates grown for 10 days on carnation leaf agar (CLA) produced hyaline microconidia with 0-2 septa, measuring 9.2 - 15.4 by 4.5 - 6.5 μm (n = 40) and hyaline macroconidia with three septa that measured 28.4 - 53.5 by 4.5 - 9 μm (n = 40). Chlamydospores were not observed. A fragment of the translation elongation factor 1-alpha (EF1-alpha) gene was amplified by polymerase chain reaction (PCR) using the primer pair EF-1/EF-2 (O'Donnell et al. 1998) from two monosporic isolates. The sequences were registered in the NCBI GenBank under accession numbers OR878541 and OR878543 (FRESIN178 and FRESIN194). BLASTn queries of NCBI GenBank identified the sequences as F. falciforme with 98% and 100% similarity to accession numbers OQ262968 and DQ246941 respectively. Fusarium ID database also identified the sequences as F. falciforme, is a member of the F. solani species complex (FSSC). Phylogenetic analysis revealed the partial EF1 sequences grouped with F. falciforme (Fig. 2). A pathogenicity test was performed on thirty strawberry plants (cv. Cabrillo) grown in sterile vermiculite. The plants were inoculated by immersing roots in 20 mL of a conidial suspension (1 × 105 conidia/mL) of isolate FRESIN194. Twelve uninoculated plants served as the control. All plants were grown for 60 days under greenhouse conditions (28 to 35°C). The assay was repeated twice. After 50 days, symptoms of root rot and wilting leaves like those observed in the field were observed. Uninoculated control plants did not develop symptoms. The fungus was reisolated from necrotic tissues of the inoculated plants and identified as F. falciforme by sequencing the EF1-alpha gene and morphological characteristics, completing Koch's postulates. Fusarium falciforme has been reported as the causal agent of root rot, stem rot, and wilt of tomato, papaya, chickpea, onion, common bean, and maize in Mexico (Díaz-Najera et al. 2021, Douriet-Angulo et al. 2021, Felix et al 2022, Tirado-Ramírez et al 2018, Vega-Gutiérrez et al. 2019a, Vega-Gutiérrez et al. 2019b). To our knowledge this is the first report of F. falciforme causing root rot and wilt on strawberry in Sinaloa, Mexico. This result provides useful information for the development and implementation of disease control strategies to mitigate damage caused by F. falciforme.

Klebsiella oxytoca inhibits Salmonella infection through multiple microbiota-context-dependent mechanisms

The Klebsiella oxytoca species complex is part of the human microbiome, especially during infancy and childhood. K. oxytoca species complex strains can produce enterotoxins, namely, tilimycin and tilivalline, while also contributing to colonization resistance (CR). The relationship between these seemingly contradictory roles is not well understood. Here, by coupling ex vivo assays with CRISPR-mutagenesis and various mouse models, we show that K. oxytoca provides CR against Salmonella Typhimurium. In vitro, the antimicrobial activity against various Salmonella strains depended on tilimycin production and was induced by various simple carbohydrates. In vivo, CR against Salmonella depended on toxin production in germ-free mice, while it was largely toxin-independent in mice with residual microbiota. This was linked to the relative levels of toxin-inducing carbohydrates in vivo. Finally, dulcitol utilization was essential for toxin-independent CR in gnotobiotic mice. Together, this demonstrates that nutrient availability is key to both toxin-dependent and substrate-driven competition between K. oxytoca and Salmonella.

Mapping of Repetitive Sequences in Brachyhypopomus brevirostris (Hypopomidae, Gymnotiformes) from the Brazilian Amazon.

Brachyhypopomus (Hypopomidae, Gymnotiformes) is a monophyletic genus consisting of 28 formally described species. Karyotypic data are available for 12 species. The same karyotype is described for two species (B. brevirostris and B. hamiltoni), as well as different karyotypes for the same species from distinct locations (B. brevirostris). In this context, B. brevirostris may constitute a cryptic species complex. Thus, in the present study, we analyzed the karyotype of B. brevirostris, from Santarém, Pará, and Tefé, Amazonas, using classical cytogenetics (conventional staining and C-banding) and molecular techniques (fluorescence in situ hybridization using 18S rDNA, 5S rDNA, U2 snRNA, and telomeric probes). The results show that samples from both locations present 2n = 38, with all chromosomes being acrocentric (FC = 38a). In both populations, 18S rDNA sequences are present on only one pair of homologous chromosomes and telomeric sequences occur only at the ends of the chromosomes. In the Tefé sample, the 5S rDNA occurs in two pairs, and the U2 snRNA in three pairs. These results are the first descriptions of these sequences for B. brevirostris samples from the Tefé locality, as well as the first karyotypic description for the Santarém locality. Future cytotaxonomic studies of this genus can benefit from these results.

Integrative taxonomy of an East Asian songbird indicates rapid dwarfism after island colonization

AbstractAnimals that colonize islands often undergo significant evolutionary changes in comparison with their continental counterparts as a response to specific island conditions. The pace of such changes can be relatively fast, which poses challenges in the evaluation of the taxonomic status of island taxa. The Japanese and Manchurian Bush Warbler species complex (Horornis diphone–canturians), which breeds in East Asia and the Japanese Archipelago, is such an avian example. This species complex exhibits significant morphological differentiation between different taxa, and the taxonomic status and phylogenetic relationships within the complex are debated. Here, we updated the taxonomy of this species complex and shed insight on its evolutionary history using multi‐locus phylogeographic and acoustic analyses. Our results support the conventional treatment of the two continental taxa borealis and canturians as subspecies of H. canturians, contrary to some recent proposals that they are affiliated to H. diphone. We also document a reduction in body size, that is dwarfism, and vocal divergence in the nominate subspecies H. d. diphone, which is endemic to the remote and isolated Ogasawara Islands. These changes may have happened following colonization of these islands, which was estimated to have taken place approximately 0.2 million years ago. Although H. d. diphone is clearly distinctive and deserves recognition as an evolutionarily significant unit, H. d. diphone and other H. diphone samples were not reciprocally monophyletic. Because of this lack of reciprocal monophyly and a relatively recent divergence time, we advocate maintaining its current subspecies status. We also detected reduced genetic diversity, measured as heterozygosity, in H. d. diphone. We suggest that conservation efforts in the Ogasawara Islands should prioritize the protection of this endemic subspecies. Collectively, our findings suggest that the separation between the populations on the East Asian continent and the Japanese Archipelago, followed by colonization of remote oceanic islands through long‐distance dispersal, underlie rapid phenotypic and genetic diversification of the Horornis diphone–canturians species complex.

Biological and molecular characterization of pydiflumetofen and phenamacril dual-resistant Fusarium graminearum strains.

Fusarium head blight (FHB) caused by Fusarium graminearum species complex (FGSG) remains a major challenge to cereal crops and resistance to key fungicides by the pathogen threatens control efficacy. Pydiflumetofen, a succinate dehydrogenase inhibitor, and phenamacril, a cyanoacrylate fungicide targeting myosin I, have been applied to combat this disease. Nonetheless, emergence of pydiflumetofen resistance in a subset of field isolates alongside laboratory-induced facile generation of phenamacril-resistant isolates signals a critical danger of resistance proliferation. Our study investigates the development of dual resistance to these fungicides in F. graminearum. Utilizing pydiflumetofen-resistant (PyR) and -sensitive (PyS) isolates, we obtained dual-resistant (PyRPhR) and phenamacril-resistant (PySPhR) mutants on potato sucrose agar containing phenamacril. Mutation rates for phenamacril resistance were comparable between pydiflumetofen-resistant and -sensitive isolates, implying independent pathways for resistance development. The mutants compromised in fungal growth, competitive viability and deoxynivalenol production, suggesting fitness penalties for the dual-resistant mutants. However, no cross-resistance was found with tebuconazole or fludioxonil. In addition, we characterized four critical amino acid changes (S217L, C423R, K537T, E420G) in the Myo1 that were verified to confer phenamacril resistance in F. graminearum. This research indicates the possibility of resistance development for both pydiflumetofen and phenamacril in F. graminearum and emphasizes the need for fungicide resistance management for FHB. © 2024 Society of Chemical Industry.

Identifying and Controlling Anthracnose Caused by Colletotrichum Taxa of Welsh Onion in Sanxing, Taiwan.

Leaves of Welsh onion (Allium fistulosum) are subject to various fungal diseases such as anthracnose (Colletotrichum species) and Stemphylium leaf blight (Stemphylium vesicarium). These diseases are the main biotic limitations to Welsh onion production in northern Taiwan. From 2018 to 2020, anthracnose symptoms were observed throughout Welsh onion fields in northern Taiwan, mainly the Sanxing area. In total, 33 strains of Colletotrichum species were isolated from diseased leaves, and major causative agents were identified based on a multilocus phylogenetic analysis using four genomic regions (act, tub2, gapdh, and internal transcribed spacer). Based on this phylogeny, Colletotrichum species causing anthracnose of Welsh onion were identified as C. spaethianum (C. spaethianum species complex) and C. circinans (C. dematium species complex) in the Sanxing area, northern Taiwan. To determine and compare the pathogenicity of each species, representative fungal strains of each species were inoculated on the cultivar 'Siao-Lyu' by spraying spore suspension onto the leaf surface. Welsh onion plants were susceptible to both species, but disease incidence and severity were higher in C. spaethianum. In total, 31 fungicides were tested to determine their efficacy in reducing mycelial growth and conidial germination of representative strains of C. spaethianum and C. circinans under laboratory conditions. Five fungicides-fluazinam, metiram, mancozeb, thiram, and dithianon-effectively reduced mycelial growth and spore germination in both C. spaethianum and C. circinans. In contrast, difenoconazole and trifloxystrobin + tebuconazole, which are commonly used in Welsh onion production in northern Taiwan, mainly the Sanxing area, were ineffective. These results serve as valuable insights for growers, enabling them to identify and address the emergence of anthracnose caused by C. spaethianum and C. circinans of Welsh onion, employing fungicides with diverse modes of action. The findings of this study support sustainable management of anthracnose in Sanxing, northern Taiwan, although further field tests of the fungicides are warranted.

Novel phages of Pseudomonas syringae unveil numerous potential auxiliary metabolic genes.

Relatively few phages that infect plant pathogens have been isolated and investigated. The Pseudomonas syringae species complex is present in various environments, including plants. It can cause major crop diseases, such as bacterial canker on apricot trees. This study presents a collection of 25 unique phage genomes that infect P. syringae. These phages were isolated from apricot orchards with bacterial canker symptoms after enrichment with 21 strains of P. syringae. This collection comprises mostly virulent phages, with only three being temperate. They belong to 14 genera, 11 of which are newly discovered, and 18 new species, revealing great genetic diversity within this collection. Novel DNA packaging systems have been identified bioinformatically in one of the new phage species, but experimental confirmation is required to define the precise mechanism. Additionally, many phage genomes contain numerous potential auxiliary metabolic genes with diversified putative functions. At least three phages encode genes involved in bacterial tellurite resistance, a toxic metalloid. This suggests that viruses could play a role in bacterial stress tolerance. This research emphasizes the significance of continuing the search for new phages in the agricultural ecosystem to unravel novel ecological diversity and new gene functions. This work contributes to the foundation for future fundamental and applied research on phages infecting phytopathogenic bacteria.

Seven new species of Rinorea (Violaceae) from the Neotropics.

Over the course of revising the genus Rinorea (Violaceae) from Colombia, field observations and herbarium studies revealed seven new species. Several of the new species described here belong to species complexes that required examination of herbarium material from across the Neotropics. Each of the new species described here have oppositely arranged leaves and belong to Rinoreasect.Pubiflorae, a section restricted to the Neotropics. Two new species are segregated from the R.ovalifolia species complex: Rinoreachiribiquetensis from Chiribiquete National Park in the Colombian Amazon and Rinoreastevensii from the Orinoco River near the border of Colombia and Venezuela. Two new species are segregated from the Rinoreahirsuta species complex: Rinoreagaleanoae-bernalii and Rinoreacogolloi, both from the eastern slopes of the Andean Central Cordillera along the mid-Magdalena River Valley in Colombia. From the widely distributed R.pubiflora species complex, we segregated one new species, Rinoreacallejasii, from southeast Panama and the Chocó in Colombia. In addition to these five new taxa segregated from widely distributed species complexes, we discovered two previously unknown species with affinities to other Neotropical Rinorea. Rinoreaaymardii is described from the Alto Orinoco-Casiquiare Biosphere Reserve in Venezuela and most closely resembles R.melanodonta from Colombia. Rinoreabetancurii is segregated from R.macrocarpa and occurs in the Amazonian Regions of Colombia, Brazil, Peru and Venezuela. In this study, we provide descriptions, illustrations and distribution maps of the new species and make preliminary assessments of the risk of extinction using the IUCN Red List Categories and Criteria. We also furnish an identification key to the species of Rinoreasect.Pubiflorae in Colombia.

Spotting mistakes: Reappraisal of Spotted Drum Stellifer punctatissimus (Meek & Hildebrand, 1925) (Teleostei: Sciaenidae) reveals species misidentification trends and suggests latitudinal sexual dimorphism

A major part of the described species is understudied, falling into the Linnean shortfall. This is a major concern for cryptic species, which require integrative approaches to better evaluate their diversity. We conducted morphological analyses using specimens of Stellifer punctatissimus, S. gomezi, and S. menezesi to reassess their taxonomical identity. We evaluated the allometric and sexual components of the morphology of the Stellifer punctatissimus species complex, and tested and discussed species hypotheses. The combined evidence of our work and previous studies agrees with the current morphological hypothesis of three species, as opposed to the two-lineage molecular hypothesis. However, as cryptic species, they overlap in most their traits, especially females and juveniles. Previously unaccounted variation of allometric and sexually dimorphic characters in this species complex revealed a confounding effect that might explain past and current taxonomic errors. Taxonomical practice of using body depth as a diagnostic character had led to juveniles and females being, respectively, described as a different species or wrongfully identified. Hence, taxonomical studies demand better assessment of allometric and sexual dimorphism components. Herein, we present new characters in a key to the Atlantic species of Stellifer, which disclose size and sexual variation unnoticed in previous studies. The contrasting growth patterns among these species could imply distinct habitat use. As a result, it could be suggested that such species are under different threats, which highlights the need of differential management and conservation strategies.

Integrative taxonomy of the groundwater amphipod Niphargus bihorensis Schellenberg, 1940 reveals a species-rich clade

Abstract The genus Niphargus is the most diverse subterranean amphipod genus in the western Palearctic region but, owing to the presence of cryptic species and homoplasy, its taxonomy and biogeographic scenarios are complex, making molecular methods essential to understand its evolution. We conducted a study combining dna-based taxonomy with traditional morphotaxonomy to investigate Niphargus bihorensis Schellenberg, 1940, known from the Western Alps and Carpathians. We redescribed the type material of N. bihorensis from Bihor County, Romania, and revealed the presence of a cryptic species, N. absconditus n. sp., in the same karstic area (Pădurea Craiului Mountains). Additionally, the Alpine populations previously attributed to N. bihorensis turned out to belong to a new, not so closely related species, N. tizianoi n. sp. Phylogenetic analyses based on a concatenated dataset of one mitochondrial and two nuclear markers suggest that the N. bihorensis species complex belongs to a strongly supported clade, together with several species distributed from Switzerland to Iran.

Genetic bias in repeated evolution of pigment loss in cave populations of the Asellus aquaticus species complex.

Similar phenotypes can evolve repeatedly under the same evolutionary pressures. A compelling example is the evolution of pigment loss and eye loss in cave-dwelling animals. While specific genomic regions or genes associated with these phenotypes have been identified in model species, it remains uncertain whether a bias towards particular genetic mechanisms exists. An isopod crustacean, Asellus aquaticus, is an ideal model organism to investigate this phenomenon. It inhabits surface freshwaters throughout Europe but has colonized groundwater on multiple independent occasions and evolved several cave populations with distinct ecomorphology. Previous studies have demonstrated that three different cave populations utilized common genetic regions, potentially the same genes, in the evolution of pigment and eye loss. Expanding on this, we conducted analysis on two additional cave populations, distinct either phylogenetically or biogeographically from those previously examined. We generated F2 hybrids from cave × surface crosses and tested phenotype-genotype associations, as well as conducted complementation tests by crossing individuals from different cave populations. Our findings revealed that pigment loss and orange eye pigment in additional cave populations were associated with the same genomic regions as observed in the three previously tested cave populations. Moreover, the lack of complementation across all cross combinations suggests that the same gene likely drives pigment loss. These results substantiate a genetic bias in the recurrent evolution of pigment loss in this model system. Future investigations should focus on the cause behind this bias, possibly arising from allele recruitment from ancestral surface populations' genetic variation or advantageous allele effects via pleiotropy.

Integrative taxonomy sheds light on species boundaries and relationships in the inbred haplodiploid ambrosia beetle genus Xyleborus

Abstract This study investigated the relationship between genetic and morphological evidence in a species complex of haplodiploid, near-inbred ambrosia beetles, which are also vectors of an invasive tree pathogen: Xyleborus ferrugineus, Xyleborus bispinatus, and Xyleborus impressus. The unorthodox genetic system may blur species boundaries and challenges the applicability of standard species concepts. Haplo-diploid scolytine systematics is rife with contentious species delimitation, but species limits have rarely been tested empirically. To test the congruence of morphology, genetics, and geography in species identity, we analyzed 16 morphological characters and two DNA barcodes (COI mtDNA and CAD DNA sequences) in samples from across Florida. Morphological characters used to distinguish species corresponded with phylogenetic relationships in the majority of individuals. The maximum intraspecific divergence for the COI barcoding region was up to 19%, among the largest reported for animals. The phylogenetic and morphological analyses suggest that the three species are valid, but the astonishing intraspecific genetic diversity suggests either additional cryptic species or pseudogenes.

Consideration of the Disease Complexes, the Missing Link to Correctly Analyze the Impact of Intercropping on Disease Development

Diversification at the plot level, through the use of intercropping (mixtures of crops), is an alternative to the conventional system of intensive agriculture, based on monospecific, usually single-variety canopies. Intercropping has been shown to provide benefits in terms of disease control. However, competition phenomena and the heterogeneity of the associated crops raise new ecological questions, particularly with regard to the dynamics and evolution of parasite populations. No study has assessed the potential impact of these associations on the dynamics of pathogenic species complexes. Changes in the nutritional status of plants and therefore in their physiological susceptibility to infection within intercropping systems could contribute to an increased diversity of ecological niches and thus affect the composition of the parasitic complex and its spatiotemporal dynamics. In this review, focusing on foliar diseases of fungal origin, and after outlining some elements of the biology and epidemiology of these fungal diseases, we will (i) describe the mechanisms that contribute to the composition of disease clusters and that drive interactions, but we will also review the strategies that these foliar diseases have adopted to deal with these co-infections; (ii) define how intercropping can lead to changes in epidemic dynamics, in particular by presenting the mechanisms that have a direct and indirect effect on disease evolution; and (iii) present the approach that should be adopted to properly study intercropping correctly in a multi-infection situation.

Phylogeny and species delimitation of ciliates in the genus Spirostomum (Class, Heterotrichea) using single-cell transcriptomes.

Ciliates are single-celled microbial eukaryotes that diverged from other eukaryotic lineages over a billion years ago. The extensive evolutionary timespan of ciliate has led to enormous genetic and phenotypic changes, contributing significantly to their high level of diversity. Recent analyses based on molecular data have revealed numerous cases of cryptic species complexes in different ciliate lineages, demonstrating the need for a robust approach to delimit species boundaries and elucidate phylogenetic relationships. Heterotrich ciliate species of the genus Spirostomum are abundant in freshwater and brackish environments and are commonly used as biological indicators for assessing water quality. However, some Spirostomum species are difficult to identify due to a lack of distinguishable morphological characteristics, and the existence of cryptic species in this genus remains largely unexplored. Previous phylogenetic studies have focused on only a few loci, namely the ribosomal RNA genes, alpha-tubulin, and mitochondrial CO1. In this study, we obtained single-cell transcriptome of 25 Spirostomum species populations (representing six morphospecies) sampled from South Korea and the USA, and used concatenation- and coalescent-based methods for species tree inference and delimitation. Phylogenomic analysis of 37 Spirostomum populations and 265 protein-coding genes provided a robustious insight into the evolutionary relationships among Spirostomum species and confirmed that species with moniliform and compact macronucleus each form a distinct monophyletic lineage. Furthermore, the multispecies coalescent (MSC) model suggests that there are at least nine cryptic species in the Spirostomum genus, three in S. minus , two in S. ambiguum, S. subtilis , and S. teres each. Overall, our fine sampling of closely related Spirostomum populations and wide scRNA-seq allowed us to demonstrate the hidden crypticity of species within the genus Spirostomum , and to resolve and provide much stronger support than hitherto to the phylogeny of this important ciliate genus.