Obligate Parasite Research Articles

Development of Aster Yellows on Crop and Noncrop Species from the Canadian Prairies

Aster yellows phytoplasmas (AYp) are a group of obligate parasites that infect a wide range of plant species, including crops such as canola and cereals and noncrops such as dandelion and wild mustard. In the Canadian Prairies, these microorganisms are mainly transmitted by a migratory species of leafhopper ( Macrosteles quadrilineatus Forbes). Although a low incidence of the disease associated with this pathogen has been reported for most years in canola fields, several outbreaks have been documented in this region. A selection of crop and noncrop species commonly found in the Canadian Prairies and Arabidopsis thaliana were used to assess the suitability of these plant species as hosts for AYp (16SrI-B). Symptom expression and phytoplasma levels were examined at different time points following exposure to infective insects. A. thaliana, barley, and canola were susceptible to infection with AYp, yet symptoms differed among these plant species. A. thaliana and canola exhibited symptoms of infection as early as 2 weeks following exposure to infected insects, whereas symptoms in barley were observed at 5 weeks. A lower incidence rate was observed in wheat, and levels of AYp in phytoplasma-infected wheat plants were low. Dandelion and sowthistle tested negative for the presence of AYp at all time points, suggesting that these are unsuitable hosts for these microorganisms. Moreover, we observed a partial disassociation between the plant species that were suitable hosts for AYp and those that had been characterized as more suitable or suitable hosts for aster leafhopper oviposition and nymphal development in previous studies. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Genomes and transcriptomes help unravel the complex life cycle of the blastoclad fungus, Coelomomyces lativittatus, an obligate parasite of mosquitoes and microcrustaceans

ABSTRACT Species of the phylum Blastocladiomycota, early-diverging zoosporic (flagellated) lineages of fungi, are vastly understudied. This phylum includes the genus Coelomomyces, which consists of more than 80 fungal species that are obligate parasites of arthropods. Known Coelomomyces species lack a complete asexual life cycle, instead surviving through an obligate heterecious alternation of generations life cycle. Despite their global distribution and interesting life cycle, little is known about the genomics of any Coelomomyces species. To address this, we generated three draft-level genomes and annotations for C. lativittatus representing its haploid meiospore, orange gamete, and amber gamete life stages. These draft genome assemblies ranged in size from 5002 to 5799 contigs, with a total length of 19.8–22.8 Mb and a mean of 7416 protein-coding genes. We then demonstrated the utility of these genomes by combining the draft annotations as a reference for analysis of C. lativittatus transcriptomes. We analyzed transcriptomes from across host-associated life stages, including infected larvae and excised mature sporangia from the mosquito Anopheles quadrimaculatus. We identified differentially expressed genes and enriched GO terms both across and within life stages and used these to make hypotheses about C. lativittatus biology. Generally, we found the C. lativittatus transcriptome to be a complex and dynamic expression landscape; GO terms related to metabolism and transport processes were enriched during infection and terms related to dispersal were enriched during sporulation. We further identified five high mobility group (HMG)-box genes in C. lativittatus, three belonging to clades with mating type (MAT) loci from other fungi, as well as four ortholog expansions in C. lativittatus compared with other fungi. The C. lativittatus genomes and transcriptomes reported here are a valuable resource and may be leveraged toward furthering understanding of the biology of these and other early-diverging fungal lineages.

Comparative analysis of the effect of nano‑copper and Cu2O treatment on Cryptocaryon irritans tomonts in large yellow croaker (Larimichthys crocea) farming

Cryptocarya irritans is an obligate parasite causing the “white spot disease.” This disease often leads to the death of a large number of cultured marine fish. This study aimed to compare the effects of cuprous oxide (Cu2O) and nano‑copper on the morphology and hatchability of tomonts to investigate the effects of copper derivatives on C. irritans tomonts. The potential mechanistic difference was also explored using the transcriptome. The results showed that Cu2O seriously damaged the internal cell structure of tomonts, and the hatching rate of the tomonts in the Cu2O-treated group was lower than that in the control group within 10 days. On the contrary, the effect of nano‑copper on tomonts was not significant. The cytoplasm of tomonts treated with nano‑copper dissolved, the internal structure was slightly loose, and the hatching rate of tomonts was even slightly higher than control group within 5 days. In the transcriptome analysis, 850 differentially expressed genes (DEGs), including 332 upregulated genes and 518 downregulated genes, were identified in the Cu2O-treated groups. However, only 56 DEGs, including 34 upregulated genes and 22 downregulated genes, were identified in the nano‑copper-treated groups. This indicated that the effect of Cu2O on tomonts was greater than nano‑copper. In the enrichment pathway analysis, DEGs in the Cu2O-treated group were mostly enriched in the ribosome and apoptosis pathways. Further analysis showed that genes (RPL18e, RPLP0, RPL10, RPL17, RPL26, RPL35, and RPL37) related to cell stress and defense were significantly upregulated and those (P53 and PKA) related to cell proliferation and development were downregulated, indicating that the killing effect was mainly mediated by different ribosomal and structural proteins. However, the DEGs (KCNMA1, CACNA1C, CNGA1) in the nano‑copper-treated group were enriched in the insulin secretion and cAMP signaling pathways, which were activate part of tomonts' innate immune response, completely different from those in the Cu2O-treated group. The results provided more information on the mechanism of killing parasites by copper derivatives.

Advances in new target molecules against schistosomiasis: A comprehensive discussion of physiological structure and nutrient intake.

Schistosomiasis, a severe parasitic disease, is primarily caused by Schistosoma mansoni, Schistosoma japonicum, or Schistosoma haematobium. Currently, praziquantel is the only recommended drug for human schistosome infection. However, the lack of efficacy of praziquantel against juvenile worms and concerns about the emergence of drug resistance are driving forces behind the research for an alternative medication. Schistosomes are obligatory parasites that survive on nutrients obtained from their host. The ability of nutrient uptake depends on their physiological structure. In short, the formation and maintenance of the structure and nutrient supply are mutually reinforcing and interdependent. In this review, we focus on the structural features of the tegument, esophagus, and intestine of schistosomes and their roles in nutrient acquisition. Moreover, we introduce the significance and modes of glucose, lipids, proteins, and amino acids intake in schistosomes. We linked the schistosome structure and nutrient supply, introduced the currently emerging targets, and analyzed the current bottlenecks in the research and development of drugs and vaccines, in the hope of providing new strategies for the prevention and control of schistosomiasis.

Phylogeography and host‐shift speciation in the vampirecups, an enigmatic clade of endophytic holoparasitic plants

AbstractAimDiversification of obligate parasites is often closely associated with historical shifts among host species. This phenomenon is well‐documented in animal parasites but still poorly understood for parasitic plants. We studied the associations between host shifts, biogeography and diversification in Cytinus, a genus of endophytic holoparasitic plants which show a remarkable disjunct distribution.LocationMediterranean Basin, southern Africa, Madagascar.TaxonCytinus.MethodsWe sampled numerous populations in South Africa and the western Mediterranean Basin, as well as a few populations in Madagascar where the genus is rare. We amplified two nuclear (ITS and SSU) and two mitochondrial (matR and nad1) loci, to infer its evolutionary history using maximum likelihood and time‐calibrated Bayesian analyses, to elucidate the factors responsible for its current distribution and to assess whether host switching is a main driver of speciation.ResultsAll genes recover lineages corresponding to the three geographic areas of Cytinus and sublineages associated with specific host plants, although they disagree on their deep‐level relationships. Ribosomal nuclear genes support an initial split of the Malagasy lineage and an Afrotropical origin of Cytinus before colonization of warm temperate latitudes. Mitochondrial genes support an initial split of the Mediterranean and Afro‐Malagasy lineages and a later split between the South African and Malagasy clades. Introgressive hybridization is the most likely explanation for this discrepancy, while horizontal gene transfers from hosts is less likely. We found strong support for cryptic host‐specific diversity, with five and eight host races in the Mediterranean and South Africa respectively, and a strong phylogenetic signal in the host–parasite associations suggestive of parasite diversification via phylogenetic host tracking.Main ConclusionsThese dated phylogenies indicate that radiation through long‐distance dispersal events and adaptation to new hosts best explain the disjunct distribution of Cytinus. We emphasize the importance of host specificity and host‐switching in the diversification and biogeography of plant endoparasites.

Unique asymmetric distribution of phosphatidylserine and phosphatidylethanolamine in Toxoplasma gondii revealed by nanoscale analysis.

Toxoplasma gondii is a highly prevalent obligate apicomplexan parasite that is important in clinical and veterinary medicine. It is known that glycerophospholipids phosphatidylserine (PtdSer) and phosphatidylethanolamine (PtdEtn), especially their expression levels and flip-flops between cytoplasmic and exoplasmic leaflets, in the membrane of T. gondii play important roles in efficient growth in host mammalian cells, but their distributions have still not been determined because of technical difficulties in studying intracellular lipid distribution at the nanometer level. In this study, we developed an electron microscopy method that enabled us to determine the distributions of PtdSer and PtdEtn in individual leaflets of cellular membranes by using quick-freeze freeze-fracture replica labeling. Our findings show that PtdSer and PtdEtn are asymmetrically distributed, with substantial amounts localized at the luminal leaflet of the inner membrane complex (IMC), which comprises flattened vesicles located just underneath the plasma membrane (see Figs. 2B and 7). We also found that PtdSer was absent in the cytoplasmic leaflet of the inner IMC membrane, but was present in considerable amounts in the cytoplasmic leaflet of the middle IMC membrane, suggesting a barrier-like mechanism preventing the diffusion of PtdSer in the cytoplasmic leaflets of the two membranes. In addition, the expression levels of both PtdSer and PtdEtn in the luminal leaflet of the IMC membrane in the highly virulent RH strain were higher than those in the less virulent PLK strain. We also found that the amount of glycolipid GM3, a lipid raft component, was higher in the RH strain than in the PLK strain. These results suggest a correlation between lipid raft maintenance, virulence, and the expression levels of PtdSer and PtdEtn in T. gondii.

Soil Aspergillus Species, Pathogenicity and Control Perspectives.

Five Aspergillus sections have members that are established agricultural pests and producers of different metabolites, threatening global food safety. Most of these pathogenic Aspergillus species have been isolated from almost all major biomes. The soil remains the primary habitat for most of these cryptic fungi. This review explored some of the ecological attributes that have contributed immensely to the success of the pathogenicity of some members of the genus Aspergillus over time. Hence, the virulence factors of the genus Aspergillus, their ecology and others were reviewed. Furthermore, some biological control techniques were recommended. Pathogenic effects of Aspergillus species are entirely accidental; therefore, the virulence evolution prediction model in such species becomes a challenge, unlike their obligate parasite counterparts. In all, differences in virulence among organisms involved both conserved and species-specific genetic factors. If the impacts of climate change continue, new cryptic Aspergillus species will emerge and mycotoxin contamination risks will increase in all ecosystems, as these species can metabolically adjust to nutritional and biophysical challenges. As most of their gene clusters are silent, fungi continue to be a source of underexplored bioactive compounds. The World Soil Charter recognizes the relevance of soil biodiversity in supporting healthy soil functions. The question of how a balance may be struck between supporting healthy soil biodiversity and the control of toxic fungi species in the field to ensure food security is therefore pertinent. Numerous advanced strategies and biocontrol methods so far remain the most environmentally sustainable solution to the control of toxigenic fungi in the field.

Detection of Secreted Effector Proteins from Phelipanche aegyptiaca During Invasion of Melon Roots.

Parasites can interact with their host plants through the induction and delivery of secreted effector proteins that facilitate plant colonization by decomposing plant cell walls and inhibiting plant immune response to weaken the defense ability of the host. Yet effectors mediating parasitic plant-host interactions are poorly understood. Phelipanche aegyptiaca is an obligate root parasite plant causing severe yield and economic losses in agricultural fields worldwide. Host resistance against P. aegyptiaca occurred during the attachment period of parasitism. Comparative transcriptomics was used to assess resistant and susceptible interactions simultaneously between P. aegyptiaca and two contrasting melon cultivars. In total, 2,740 secreted proteins from P. aegyptiaca were identified here. Combined with transcriptome profiling, 209 candidate secreted effector proteins (CSEPs) were predicted, with functional annotations such as cell wall degrading enzymes, protease inhibitors, transferases, kinases, and elicitor proteins. A heterogeneous expression system in Nicotiana benthamiana was used to investigate the functions of 20 putatively effector genes among the CSEPs. Cluster 15140.0 can suppress BAX-triggered programmed cell death in N. benthamiana. These findings showed that the prediction of P. aegyptiaca effector proteins based on transcriptomic analysis and multiple bioinformatics software is effective and more accurate, providing insights into understanding the essential molecular nature of effectors and laying the foundation of revealing the parasite mechanism of P. aegyptiaca, which is helpful in understanding parasite-host plant interaction.

Decision rules for egg-color-based rejection by two cavity-nesting hosts of the brown-headed cowbird.

Hosts of obligate avian brood parasites often evolve defense mechanisms to avoid rearing unrelated young. One common defense is egg rejection, for which hosts often rely on eggshell color. Most research has assumed that hosts respond to perceived color differences between their own eggs and parasite eggs regardless of the particular color; however, recent experiments have found that many hosts respond more strongly to brown foreign eggs than to equally dissimilar blue eggs. Yet, none of these prior studies tested a brown-egg-laying species and, with only one exception, all were conducted in open nests where light levels are considered sufficient for effective color-based egg discrimination. Here, we explored how two cavity-nesting hosts of the parasitic brown-headed cowbird (Molothrus ater) - the blue-egg-laying eastern bluebird (Sialia sialis) and the brown-egg-laying house wren (Troglodytes aedon) - respond to experimental eggs painted six distinct colors ranging from blue to brown. Rejection responses of both hosts were best predicted by perceived differences in color between the model egg and their own eggs. Specifically, we found that house wrens preferentially rejected eggs bluer than their own eggs. However, although we found that bluebirds relied on perceived differences in color for their egg rejection decisions, further tests are needed to determine whether they preferentially rejected brown eggs or simply responded to absolute perceived differences in color. These findings demonstrate that these cavity-nesting birds treat perceived color differences in distinct ways, which has important implications on the coevolutionary arms races and the interpretation of avian-perceived color differences.

Widespread mermithid nematode parasitism of Cretaceous insects.

Mermithid nematodes are obligate invertebrate parasites dating back to the Early Cretaceous. Their fossil record is sparse, especially before the Cenozoic, thus little is known about their early host associations. This study reports 16 new mermithids associated with their insect hosts from mid-Cretaceous Kachin amber, 12 of which include previously unknown hosts. These fossils indicate that mermithid parasitism of invertebrates was already widespread and played an important role in the mid-Cretaceous terrestrial ecosystem. Remarkably, three hosts (bristletails, barklice, and perforissid planthoppers) were previously unknown to be parasitized by mermithids both past and present. Furthermore, our study shows that in contrast to their Cenozoic counterparts, Cretaceous nematodes including mermithids are more abundant in non-holometabolous insects. This result suggests that nematodes had not completely exploited the dominant Holometabola as their hosts until the Cenozoic. This study reveals what appears to be a vanished history of nematodes that parasitized Cretaceous insects.

Targeting proteases and proteolytic processing of unusual N-terminal extensions of Plasmodium proteins: parasite peculiarity

More than sesquicentennial years of malarial research, however the unique malarial parasite, Plasmodium still bewilders us with its atypical characteristic features. Elimination strategies, deeper knowledge of the parasite biology and pathways can help combat this global health concern that affects ∼250 million people annually. In this review, we unveil an unusual phenomenon observed in the parasite proteome, N-terminal extensions in proteins and highlight that the proteases that may be involved in their processing events, are potential candidates to target this pathogen. Plasmodium encodes larger proteins as compared to its eukaryotic counterparts with homology regions present in the C-terminus of the protein. In contrast, the function of unusual extensions in the N-terminus remains mostly elusive. This novelty observed in Plasmodium proteins is collated here with a focus on replication proteins. The plausible functions and prevalence of these extensions, despite the reduction in genome size, through the parasite evolution are also mentioned. We hypothesize that these extensions, propagated via the energy consuming cellular processes in the otherwise host-dependent obligate parasite, are beneficial to the parasite in ways that are yet to be explored. Consequently, targeting the proteolytic processing of these proteins and the involved proteases would serve as a new drug development regimen to tackle the emerging resistance in parasites to existing antimalarials.

Root-knot nematodes produce functional mimics of tyrosine-sulfated plant peptides

Root-knot nematodes (Meloidogyne spp.) are highly evolved obligate parasites threatening global food security. These parasites have a remarkable ability to establish elaborate feeding sites in roots, which are their only source of nutrients throughout their life cycle. A wide range of nematode effectors have been implicated in modulation of host pathways for defense suppression and/or feeding site development. Plants produce a diverse array of peptide hormones including PLANT PEPTIDE CONTAINING SULFATED TYROSINE (PSY)-family peptides, which promote root growth via cell expansion and proliferation. A sulfated PSY-like peptide RaxX (required for activation of XA21 mediated immunity X) produced by the biotrophic bacterial pathogen (Xanthomonas oryzae pv. oryzae) has been previously shown to contribute to bacterial virulence. Here, we report the identification of genes from root-knot nematodes predicted to encode PSY-like peptides (MigPSYs) with high sequence similarity to both bacterial RaxX and plant PSYs. Synthetic sulfated peptides corresponding to predicted MigPSYs stimulate root growth in Arabidopsis. MigPSY transcript levels are highest early in the infection cycle. Downregulation of MigPSY gene expression reduces root galling and egg production, suggesting that the MigPSYs serve as nematode virulence factors. Together, these results indicate that nematodes and bacteria exploit similar sulfated peptides to hijack plant developmental signaling pathways to facilitate parasitism.

Drivers and consequences of bacteriophage host range.

Bacteriophages are obligate parasites of bacteria characterized by the breadth of hosts that they can infect. This "host range" depends on the genotypes and morphologies of the phage and the bacterial host, but also on the environment in which they are interacting. Understanding phage host range is critical to predicting the impacts of these parasites in their natural host communities and their utility as therapeutic agents, but is also key to predicting how phages evolve and in doing so drive evolutionary change in their host populations, including through movement of genes among unrelated bacterial genomes. Here, we explore the drivers of phage infection and host range from the molecular underpinnings of the phage-host interaction to the ecological context in which they occur. We further evaluate the importance of intrinsic, transient, and environmental drivers shaping phage infection and replication, and discuss how each influences host range over evolutionary time. The host range of phages has great consequences in phage-based application strategies, as well as natural community dynamics, and we therefore highlight both recent developments and key open questions in the field as phage-based therapeutics come back into focus.

An insight into the functional genomics and species classification of Eudiplozoon nipponicum (Monogenea, Diplozoidae), a haematophagous parasite of the common carp Cyprinus carpio

BackgroundMonogenea (Platyhelminthes, Neodermata) are the most species-rich class within the Neodermata superclass of primarily fish parasites. Despite their economic and ecological importance, monogenean research tends to focus on their morphological, phylogenetic, and population characteristics, while comprehensive omics analyses aimed at describing functionally important molecules are few and far between. We present a molecular characterisation of monogenean representative Eudiplozoon nipponicum, an obligate haematophagous parasite infecting the gills of the common carp. We report its nuclear and mitochondrial genomes, present a functional annotation of protein molecules relevant to the molecular and biochemical aspect of physiological processes involved in interactions with the fish hosts, and re-examinate the taxonomic position of Eudiplozoon species within the Diplozoidae family.ResultsWe have generated 50.81 Gbp of raw sequencing data (Illumina and Oxford Nanopore reads), bioinformatically processed, and de novo assembled them into a genome draft 0.94 Gbp long, consisting of 21,044 contigs (N50 = 87 kbp). The final assembly represents 57% of the estimated total genome size (~ 1.64 Gbp), whereby repetitive and low-complexity regions account for ~ 64% of the assembled length. In total, 36,626 predicted genes encode 33,031 proteins and homology-based annotation of protein-coding genes (PCGs) and proteins characterises 14,785 (44.76%) molecules. We have detected significant representation of functional proteins and known molecular functions. The numbers of peptidases and inhibitors (579 proteins), characterised GO terms (16,016 unique assigned GO terms), and identified KEGG Orthology (4,315 proteins) acting in 378 KEGG pathways demonstrate the variety of mechanisms by which the parasite interacts with hosts on a macromolecular level (immunomodulation, feeding, and development). Comparison between the newly assembled E. nipponicum mitochondrial genome (length of 17,038 bp) and other diplozoid monogeneans confirms the existence of two distinct Eudiplozoon species infecting different fish hosts: Cyprinus carpio and Carassius spp.ConclusionsAlthough the amount of sequencing data and characterised molecules of monogenean parasites has recently increased, a better insight into their molecular biology is needed. The E. nipponicum nuclear genome presented here, currently the largest described genome of any monogenean parasite, represents a milestone in the study of monogeneans and their molecules but further omics research is needed to understand these parasites’ biological nature.

Food Provisioning and Body Mass of Nestling Meadow Pipits and Cuckoos

Common Cuckoos Cuculus canorus are obligate brood parasites, laying eggs in nests of other species. Cuckoo nestlings are often thought to be insatiable, compared to host broods. However, in Reed Warblers Acrocephalus scirpaceus and a few other species, cuckoo nestlings are fed at most only as frequently as a host brood. To add to the small body of knowledge on feeding frequencies and body mass development of Cuckoo nestlings, I studied Cuckoos using another host, the Meadow Pipit Anthus pratensis, in the Dutch dunes (2019–2021). By filming feeding parents (hosts) I determined the provisioning frequency for broods of Pipits and for nestling Cuckoos. I found that nestling Cuckoos receive on average fewer feeds per hour than broods of Meadow Pipits. Furthermore, overall Cuckoos received as many feeds during their 22-day nestling period as a brood of Pipits during their 13 days in the nest. At 13 days of age (day 1 is the day of hatching), the single Cuckoo was as heavy as a brood of four Meadow Pipits. At 22 days, the maximum weights of the nestling Cuckoos were greater than the Meadow Pipit broods and varied between 91 and 105 g. Thus, although Cuckoos are being fed less frequently, young Cuckoos are heavier at fledging than a whole brood of Pipits. This could be because Cuckoos are fed larger prey. Alternatively, Cuckoos may require less food because their thermoregulatory costs could be smaller: they have almost black skin which absorbs solar radiation efficiently, do not have to compete with siblings in the nest and, once older and feathered, have a smaller surface-to-volume ratio than a Pipit brood of four nestlings.

Effects of water movement and temperature on Rhizophydium infection of Planktothrix in a shallow hypereutrophic lake.

Grand Lake St. Marys (GLSM) is a popular recreational lake located in western Ohio, United States, generating nearly $150 million in annual revenue. However, recurring algal blooms dominated by Planktothrix agardhii, which can produce harmful microcystin toxins, have raised concerns about water safety and negatively impacted the local economy. Planktothrix agardhii is host to a number of parasites and pathogens, including an obligate fungal parasite in the Chytridiomycota (chytrids). In this study, we investigated the potential of these chytrid (Rhizophydium sp.) to infect P. agardhii blooms in the environment by modifying certain environmental conditions thought to limit infection prevalence in the wild. With a focus on temperature and water mixing, mesocosms were designed to either increase or decrease water flow compared to the control (water outside the mesocosm). In the control and water circulation mesocosms, infections were found infrequently and were found on less than 0.75% of the Planktothrix population. On the other hand, by decreasing the water flow to stagnation, chytrid infections were more frequent (found in nearly 3x as many samples) and more prevalent, reaching a maximum infection rate of 4.12%. In addition, qPCR coupled with 16S-18S sequencing was utilized to confirm the genetic presence of both host and parasite, as well as to better understand the effect of water circulation on the community composition. Statistical analysis of the data confirmed that chytrid infection was dependent on water temperature, with infections predominantly occurring between 19°C and 23°C. Additionally, water turbulence can significantly reduce the infectivity of chytrids, as infections were mostly found in stagnant mesocosms. Further, decreasing the water circulation promoted the growth of the cyanobacterial population, while increasing water agitation promoted the growth of green algae (Chlorophyta). This study starts to explore the environmental factors that affect chytrid pathogenesis which can provide valuable insights into controlling measures to reduce the prevalence of harmful algal blooms and improve water quality in GLSM and similarly affected waterbodies.

Contrasting outcomes of genome reduction in mikrocytids and microsporidians

BackgroundIntracellular symbionts often undergo genome reduction, losing both coding and non-coding DNA in a process that ultimately produces small, gene-dense genomes with few genes. Among eukaryotes, an extreme example is found in microsporidians, which are anaerobic, obligate intracellular parasites related to fungi that have the smallest nuclear genomes known (except for the relic nucleomorphs of some secondary plastids). Mikrocytids are superficially similar to microsporidians: they are also small, reduced, obligate parasites; however, as they belong to a very different branch of the tree of eukaryotes, the rhizarians, such similarities must have evolved in parallel. Since little genomic data are available from mikrocytids, we assembled a draft genome of the type species, Mikrocytos mackini, and compared the genomic architecture and content of microsporidians and mikrocytids to identify common characteristics of reduction and possible convergent evolution.ResultsAt the coarsest level, the genome of M. mackini does not exhibit signs of extreme genome reduction; at 49.7 Mbp with 14,372 genes, the assembly is much larger and gene-rich than those of microsporidians. However, much of the genomic sequence and most (8075) of the protein-coding genes code for transposons, and may not contribute much of functional relevance to the parasite. Indeed, the energy and carbon metabolism of M. mackini share several similarities with those of microsporidians. Overall, the predicted proteome involved in cellular functions is quite reduced and gene sequences are extremely divergent. Microsporidians and mikrocytids also share highly reduced spliceosomes that have retained a strikingly similar subset of proteins despite having reduced independently. In contrast, the spliceosomal introns in mikrocytids are very different from those of microsporidians in that they are numerous, conserved in sequence, and constrained to an exceptionally narrow size range (all 16 or 17 nucleotides long) at the shortest extreme of known intron lengths.ConclusionsNuclear genome reduction has taken place many times and has proceeded along different routes in different lineages. Mikrocytids show a mix of similarities and differences with other extreme cases, including uncoupling the actual size of a genome with its functional reduction.

Female red-winged blackbirds (Agelaius phoeniceus) do not alter nest site selection, maternal programming, or hormone-mediated maternal effects in response to perceived nest predation or brood parasitism risk

Predation or brood parasitism risks can change the behaviors and reproductive decisions in many parental animals. For oviparous species, mothers can mitigate their reproductive success in at least three ways: (1) by avoiding nest sites with high predation or parasitism risks, (2) through hormonal maternal effects that developmentally prime offspring for survival in risky environments, or (3) by investing less in reproduction when predation or parasitism risks are high. Here, we tested if perceived predation and parasitism risks can induce any of these behavioral or physiological responses by exposing female red-winged blackbirds (Agelaius phoeniceus) to playbacks of two major nest threats, a predator (Cooper’s hawk, Accipiter cooperii) and an obligate brood parasite (brown-headed cowbird; Molothrus ater), as well as two controls (harmless Eastern meadowlark, Sturnella magna; and silence). We found that female blackbirds did not avoid nesting at sites treated with predator or brood parasite playbacks, nor were females more likely to abandon nesting attempts at these sites. Egg size and yolk hormone profiles, which are common proxies for maternal investment in oviparous species, were statistically similar across treatment sites. Instead, we found intraclutch variation in yolk steroid hormone profiles: concentrations of three progestogens (pregnanedione, 17α-hydroxypregnenolone, and deoxycorticosterone) and two androgens (testosterone and androstenedione) were higher in third-laid than first-laid eggs. Our study largely confirms previous findings of consistent intraclutch yolk hormone variation in this species, in birds in general, and in other oviparous lineages, but uniquely reports on several yolk steroid hormones largely overlooked in the literature on hormone-mediated maternal effects.

Разработка ДНК-маркеров признака устойчивости подсолнечника к расе G заразихи (Orobanche cumana Wallr.)

Broomrape (Orobanche cumana Wallr.) is an obligatory parasite which is able to decrease significantly sunflower yields. The race G is the most spread and virulence broomrape race in the Russian Federation. Cultivation of genetically resistance sunflower varieties and hybrids is the most ecologic and economically efficient method to control this parasite plant. Usage of DNA-markers when selecting the resistant plants increases significantly the effectiveness of a breeding process. To mark the sunflower resistance to broomrape race G, using bioinformative approaches, we developed new DNA-markers: four SCAR (sequence characterized amplified region) and ten SSR (simple sequence repeat). These markers were tested on 20 lines and hybrids of sunflower. Due to the results of this testing, all SSR- and two SCAR-markers were eliminated by steps. SCAR-marker RORS1 was characterized with the presence of an amplified DNA fragment with a length ≈ 168 bps and showed association with a trait of resistance, and SORS1 with a fragment length ≈ 322 bps had a relation with a sus-ceptible phenotype. These markers composed a marker system for a multiplex PCR, which allowed distinguishing resistant and susceptible lines and hybrids, except a line VK678 and F1 hybrids developed using this line as parental one. To eliminate faults of the marker system, seven SCAR-markers analogous to SORS1 were created. Due to estimation result, we selected a marker SORS9 characterized with the presence an amplified DNA fragment with a length ≈ 217 bps in susceptible phenotypes. The new system of markers RORS1/SORS9 for a multiplex PCR was validated on 70 sunflower lines and hybrids. The susceptible lines and hybrids were characterized with amplification of only marker SORS9, resistant lines – only marker RORS1, and resistant hybrids – both markers. The developed marker system allowed distinguishing all susceptible lines and hybrids from the resistant sunflower genotypes.

Proteins released into the plant apoplast by the obligate parasitic protist Albugo selectively repress phyllosphere-associated bacteria.

Biotic and abiotic interactions shape natural microbial communities. The mechanisms behind microbe-microbe interactions, particularly those protein based, are not well understood. We hypothesize that released proteins with antimicrobial activity are a powerful and highly specific toolset to shape and defend plant niches. We have studied Albugo candida, an obligate plant parasite from the protist Oomycota phylum, for its potential to modulate the growth of bacteria through release of antimicrobial proteins into the apoplast. Amplicon sequencing and network analysis of Albugo-infected and uninfected wild Arabidopsis thaliana samples revealed an abundance of negative correlations between Albugo and other phyllosphere microbes. Analysis of the apoplastic proteome of Albugo-colonized leaves combined with machine learning predictors enabled the selection of antimicrobial candidates for heterologous expression and study of their inhibitory function. We found for three candidate proteins selective antimicrobial activity against Gram-positive bacteria isolated from A. thaliana and demonstrate that these inhibited bacteria are precisely important for the stability of the community structure. We could ascribe the antibacterial activity of the candidates to intrinsically disordered regions and positively correlate it with their net charge. This is the first report of protist proteins with antimicrobial activity under apoplastic conditions that therefore are potential biocontrol tools for targeted manipulation of the microbiome.