RESUMO
The endosymbiotic bacteria Spiroplasma (Mollicutes) infect diverse plants and arthropods, and some of which induce male killing, where male hosts are killed during development. Male-killing Spiroplasma strains belong to either the phylogenetically distant Citri-Poulsonii or Ixodetis groups. In Drosophila flies, Spiroplasma poulsonii induces male killing via the Spaid toxin. While Spiroplasma ixodetis infects a wide range of insects and arachnids, little is known about the genetic basis of S. ixodetis-induced male killing. Here, we analyzed the genome of S. ixodetis strains in the pea aphid Acyrthosiphon pisum (Aphididae, Hemiptera). Genome sequencing constructed a complete genome of a male-killing strain, sAp269, consisting of a 1.5 Mb circular chromosome and an 80 Kb plasmid. sAp269 encoded putative virulence factors containing either ankyrin repeat, ovarian tumor-like deubiquitinase, or ribosome inactivating protein domains, but lacked the Spaid toxin. Further comparative genomics of Spiroplasma strains in A. pisum biotypes adapted to different host plants revealed their phylogenetic associations and the diversity of putative virulence factors. Although the mechanisms of S. ixodetis-induced male killing in pea aphids remain elusive, this study underlines the dynamic genome evolution of S. ixodetis and proposes independent acquisition events of male-killing mechanisms in insects.
Assuntos
Afídeos , Genoma Bacteriano , Filogenia , Spiroplasma , Simbiose , Animais , Spiroplasma/genética , Spiroplasma/fisiologia , Spiroplasma/classificação , Afídeos/microbiologia , Masculino , Fenótipo , Genômica , Fatores de Virulência/genética , Feminino , Pisum sativum/microbiologia , Pisum sativum/parasitologiaRESUMO
Cell cultures have provided an ideal habitat for a wide variety of Mycoplasma and Acholeplasma species since the earliest days of in-vitro culture. The possibility of contamination with Spiroplasma species was addressed by Regulatory Authorities due to the increased commercial use of insect cells, recognising that Spiroplasmas have been isolated from many types of arthropod and also that insect cell cultures support Spiroplasma growth as they have been used for cultivation of fastidious species. In this study we re-examined two cell culture samples previously confirmed as contaminated with mollicutes by cultural methods. One isolate had undergone sequencing which had placed it in the S. citri phylogenetic group, whilst the other had not been identified. Using modern sequencing methods we were able to further identify both isolates to species level.
Assuntos
Técnicas de Cultura de Células , Spiroplasma , Tenericutes , Animais , Filogenia , Spiroplasma/classificação , Spiroplasma/isolamento & purificação , Tenericutes/classificação , Tenericutes/isolamento & purificaçãoRESUMO
Spiroplasma is a genus of Mollicutes whose members include plant pathogens, insect pathogens and endosymbionts of animals. Spiroplasma phenotypes have been repeatedly observed to be spontaneously lost in Drosophila cultures, and several studies have documented a high genomic turnover in Spiroplasma symbionts and plant pathogens. These observations suggest that Spiroplasma evolves quickly in comparison to other insect symbionts. Here, we systematically assess evolutionary rates and patterns of Spiroplasma poulsonii, a natural symbiont of Drosophila. We analysed genomic evolution of sHy within flies, and sMel within in vitro culture over several years. We observed that S. poulsonii substitution rates are among the highest reported for any bacteria, and around two orders of magnitude higher compared with other inherited arthropod endosymbionts. The absence of mismatch repair loci mutS and mutL is conserved across Spiroplasma, and likely contributes to elevated substitution rates. Further, the closely related strains sMel and sHy (>99.5â% sequence identity in shared loci) show extensive structural genomic differences, which potentially indicates a higher degree of host adaptation in sHy, a protective symbiont of Drosophila hydei. Finally, comparison across diverse Spiroplasma lineages confirms previous reports of dynamic evolution of toxins, and identifies loci similar to the male-killing toxin Spaid in several Spiroplasma lineages and other endosymbionts. Overall, our results highlight the peculiar nature of Spiroplasma genome evolution, which may explain unusual features of its evolutionary ecology.
Assuntos
Drosophila/microbiologia , Proteínas MutL/genética , Proteínas MutS/genética , Spiroplasma/classificação , Substituição de Aminoácidos , Animais , Proteínas de Bactérias/genética , Evolução Molecular , Taxa de Mutação , Filogenia , Análise de Sequência de DNA , Spiroplasma/genética , SimbioseRESUMO
BACKGROUND: The microbiome is an integral component of many animal species, potentially affecting behavior, physiology, and other biological properties. Despite this importance, bacterial communities remain vastly understudied in many groups of invertebrates, including mites. Quill mites (Acariformes: Syringophilidae) are a poorly known group of permanent bird ectoparasites that occupy quills of feathers and feed on bird subcutaneous tissue and fluids. Most of the known species have strongly female-biased sex ratio, and it was hypothesized that this is caused by endosymbiotic bacteria. Previously, Anaplasma phagocytophilum (Foggie) and a high diversity of Wolbachia strains were detected in quill mites via targeted PCR screens. Here, we use an unbiased 16S rRNA gene amplicon sequencing approach to determine other bacteria that potentially impact quill mite biology. RESULTS: We performed 16S rRNA gene amplicon sequencing of 126 quill mite individuals from eleven species parasitizing twelve species (four families) of passeriform birds. In addition to Wolbachia, we found Spiroplasma as potential symbiont of quill mites. Consistently, high Spiroplasma titers were only found in individuals of two mite species associated with finches of the genus Carduelis, suggesting a history of horizontal transfers of Spiroplasma via the bird host. Furthermore, there was evidence for Spiroplasma negatively affecting Wolbachia titers. We found no evidence for the previously reported Anaplasma in quill mites, but detected sequences of high similarity to the potential pathogens Brucella and Bartonella at low abundances. Other amplicon sequence variants (ASVs) could be assigned to a diverse number of bacterial taxa, including several that were previously isolated from bird skin. Further, many frequently found ASVs were assigned to taxa that show a very broad distribution with no strong prior evidence for symbiotic association with animals. We interpret these findings as evidence for a scarcity of resident microbial associates (other than inherited symbionts) in quill mites.
Assuntos
Aves/parasitologia , Plumas/parasitologia , Ácaros/microbiologia , Spiroplasma/classificação , Wolbachia/classificação , Animais , Biodiversidade , DNA Bacteriano/genética , Microbiota , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Spiroplasma/isolamento & purificação , Simbiose , Wolbachia/isolamento & purificaçãoRESUMO
Study of the mutualistic associations between facultative symbionts and aphids are developed only in a few models. That survey on the situation and distribution of the symbionts in a certain area is helpful to obtain clues for the acquisition and spread of them as well as their roles played in host evolution. To understand the infection patterns of seven facultative symbionts (Serratia symbiotica, Hamiltonella defensa, Regiella insecticola, Rickettsia, Spiroplasma, Wolbachia, and Arsenophonus) in Rhopalosiphum padi (Linnaeus) and Rhopalosiphum maidis (Fitch), we collected 882 R. maidis samples (37 geographical populations) from China and 585 R. padi samples (32 geographical populations) from China and Europe. Results showed that both species were widely infected with various symbionts and totally 50.8% of R. maidis and 50.1% of R. padi were multi-infected with targeted symbionts. However, very few Rhopalosiphum aphids were infected with S. symbiotica. The infection frequencies of some symbionts were related to the latitude of collecting sites, suggesting the importance of environmental factors in shaping the geographic distribution of facultative symbionts. Also, R. maidis and R. padi were infected with different H. defensa strains based on phylogenetic analysis which may be determined by host ×symbiont genotype interactions. According to our results, the ubiquitous symbionts may play important roles in the evolution of their host aphid and their impacts on adaptation of R. padi and R. maidis were discussed as well.
Assuntos
Afídeos/microbiologia , Gammaproteobacteria/isolamento & purificação , Filogeografia , Rickettsiales/isolamento & purificação , Spiroplasma/isolamento & purificação , Simbiose , Animais , China , Europa (Continente) , Gammaproteobacteria/classificação , Rickettsiales/classificação , Spiroplasma/classificaçãoRESUMO
The bacterium Spiroplasma ixodetis is a maternally inherited endosymbiont primarily described from ticks but also found widespread across other arthropods. While it has been identified as a male-killing agent in some insect species, the consequences of infection with S. ixodetis in ticks are entirely unknown, and it is unclear how this endosymbiont spreads across tick species. Here, we have investigated this aspect through the examination of the diversity and evolutionary history of S. ixodetis infections in 12 tick species and 12 other arthropod species. Using a multi-locus typing approach, we identified that ticks harbor a substantial diversity of divergent S. ixodetis strains. Phylogenetic investigations revealed that these S. ixodetis strains do not cluster within a tick-specific subclade but rather exhibit distinct evolutionary origins. In their past, these strains have undergone repeated horizontal transfers between ticks and other arthropods, including aphids and flies. This diversity pattern strongly suggests that maternal inheritance and horizontal transfers are key drivers of S. ixodetis spread, dictating global incidence of infections across tick communities. We do not, however, detect evidence of S. ixodetis-based male-killing since we observed that infections were widely present in both males and females across populations of the African blue tick Rhipicephalus decoloratus.
Assuntos
Artrópodes/microbiologia , Infecções por Bactérias Gram-Negativas/transmissão , Filogenia , Spiroplasma/genética , Simbiose , Carrapatos/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Transmissão de Doença Infecciosa , Feminino , Variação Genética , Transmissão Vertical de Doenças Infecciosas , Masculino , Tipagem de Sequências Multilocus , Spiroplasma/classificaçãoRESUMO
Bacterial symbionts may influence the fitness of their herbivore hosts, but such effects have been poorly studied across most invertebrate groups. The spider mite, Tetranychus truncatus, is a polyphagous agricultural pest harboring various bacterial symbionts whose function is largely unknown. Here, by using a high-throughput 16S rRNA amplicon sequencing approach, we characterized the bacterial diversity and community composition of spider mites fed on five host plants after communities were modified following tetracycline exposure. We demonstrated that spider mite bacterial diversity and community composition were significantly affected by host plants and antibiotics. In particular, the abundance of the maternally inherited endosymbionts Wolbachia and Spiroplasma significantly differed among spider mites that were reared on different plant species and were completely removed by antibiotics. There was an overall tendency for daily fecundity to be lower in the mites with reduced bacterial diversity following the antibiotic treatment. Our data suggest that host plants and antibiotics can shape spider mite bacterial communities and that bacterial symbionts improve mite performance.
Assuntos
Bactérias/isolamento & purificação , Microbiota , Tetranychidae/microbiologia , Tetranychidae/fisiologia , Animais , Bactérias/classificação , Bactérias/genética , Feminino , Fertilidade , Especificidade de Hospedeiro , Masculino , Plantas/parasitologia , Spiroplasma/classificação , Spiroplasma/genética , Spiroplasma/isolamento & purificação , Wolbachia/classificação , Wolbachia/genética , Wolbachia/isolamento & purificaçãoRESUMO
We report a disseminated infection caused by Spiroplasma apis, a honeybee pathogen, in a patient in France who had X-linked agammaglobulinemia. Identification was challenging because initial bacterial cultures and direct examination by Gram staining were negative. Unexplained sepsis in patients with agammaglobulinemia warrants specific investigation to identify fastidious bacteria such as Spiroplasma spp.
Assuntos
Agamaglobulinemia/complicações , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/etiologia , Spiroplasma , Adulto , Agamaglobulinemia/diagnóstico , Agamaglobulinemia/terapia , Antibacterianos/uso terapêutico , Biópsia , França , Doenças Genéticas Ligadas ao Cromossomo X/complicações , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Masculino , RNA Ribossômico 16S/metabolismo , Pele/microbiologia , Pele/patologia , Spiroplasma/classificação , Spiroplasma/genética , Resultado do TratamentoRESUMO
The heritable endosymbiotic bacterium Spiroplasma is found in the harlequin ladybird Harmonia axyridis. The proportion of beetles infected with Spiroplasma in different native H. axyridis populations varies from 2% to 49%. We investigated the polymorphism of Spiroplasma strains in samples from individual beetles from Kyoto, Vladivostok, Troitsa Bay, Novosibirsk, and Gorno-Altaisk. To identify Spiroplasma strains, we analyzed nucleotide polymorphisms of the 16S rRNA gene and the ribosomal internal transcribed spacer (ITS1). The majority of infected beetles were infected with two or more Spiroplasma strains. We measured Spiroplasma density in beetles with different infection status using quantitative PCR. The abundance of Spiroplasma in samples with a single infection is an order of magnitude lower than in samples with multiple infections. Density dependent biological effects of Spiroplasma are discussed.
Assuntos
Biodiversidade , Besouros/microbiologia , Spiroplasma/fisiologia , Animais , Besouros/citologia , Feminino , Espaço Intracelular/microbiologia , Larva/microbiologia , Masculino , Filogenia , Spiroplasma/classificaçãoRESUMO
Genetic differentiation among symbiotic bacteria is important in shaping biodiversity. The genus Spiroplasma contains species occupying diverse niches and is a model system for symbiont evolution. Previous studies have established that two mosquito-associated species have diverged extensively in their carbohydrate metabolism genes despite having a close phylogenetic relationship. Notably, although the commensal Spiroplasma diminutum lacks identifiable pathogenicity factors, the pathogenic Spiroplasma taiwanense was found to have acquired a virulence factor glpO and its associated genes through horizontal transfer. However, it is unclear if these acquired genes have been integrated into the regulatory network. In this study, we inferred the gene content evolution in these bacteria, as well as examined their transcriptomes in response to glucose availability. The results indicated that both species have many more gene acquisitions from the Mycoides-Entomoplasmataceae clade, which contains several important pathogens of ruminants, than previously thought. Moreover, several acquired genes have higher expression levels than the vertically inherited homologs, indicating possible functional replacement. Finally, the virulence factor and its functionally linked genes in S. taiwanense were up-regulated in response to glucose starvation, suggesting that these acquired genes are under expression regulation and the pathogenicity may be a stress response. In summary, although differential gene losses are a major process for symbiont divergence, gene gains are critical in counteracting genome degradation and driving diversification among facultative symbionts.
Assuntos
Culicidae/microbiologia , Transferência Genética Horizontal , Genes Bacterianos , Spiroplasma/genética , Simbiose , Fatores de Virulência/genética , Animais , DNA Bacteriano , Evolução Molecular , Genoma Bacteriano , Filogenia , Spiroplasma/classificação , TranscriptomaRESUMO
Profiling of wild and laboratory tsetse populations using 16S rRNA gene amplicon sequencing allowed us to examine whether the "Wigglesworthia-Sodalis-Wolbachia dogma" operates across species and populations. The most abundant taxa, in wild and laboratory populations, were Wigglesworthia (the primary endosymbiont), Sodalis and Wolbachia as previously characterized. The species richness of the microbiota was greater in wild than laboratory populations. Spiroplasma was identified as a new symbiont exclusively in Glossina fuscipes fuscipes and G. tachinoides, members of the palpalis sub-group, and the infection prevalence in several laboratory and natural populations was surveyed. Multi locus sequencing typing (MLST) analysis identified two strains of tsetse-associated Spiroplasma, present in G. f. fuscipes and G. tachinoides. Spiroplasma density in G. f. fuscipes larva guts was significantly higher than in guts from teneral and 15-day old male and female adults. In gonads of teneral and 15-day old insects, Spiroplasma density was higher in testes than ovaries, and was significantly higher density in live versus prematurely deceased females indicating a potentially mutualistic association. Higher Spiroplasma density in testes than in ovaries was also detected by fluorescent in situ hybridization in G. f. fuscipes.
Assuntos
Enterobacteriaceae/isolamento & purificação , Spiroplasma/isolamento & purificação , Moscas Tsé-Tsé/microbiologia , Moscas Tsé-Tsé/parasitologia , Wigglesworthia/isolamento & purificação , Wolbachia/isolamento & purificação , Animais , Animais Selvagens/microbiologia , Animais Selvagens/parasitologia , Enterobacteriaceae/classificação , Enterobacteriaceae/genética , Enterobacteriaceae/fisiologia , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Masculino , Tipagem de Sequências Multilocus , Ovário/microbiologia , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de RNA , Especificidade da Espécie , Spiroplasma/classificação , Spiroplasma/genética , Spiroplasma/fisiologia , Simbiose , Testículo/microbiologia , Distribuição Tecidual , Moscas Tsé-Tsé/classificação , Moscas Tsé-Tsé/crescimento & desenvolvimento , Wigglesworthia/classificação , Wigglesworthia/genética , Wigglesworthia/fisiologia , Wolbachia/classificação , Wolbachia/genética , Wolbachia/fisiologiaRESUMO
Cotylorhiza tuberculata is an important scyphozoan jellyfish producing population blooms in the Mediterranean probably due to pelagic ecosystem's decay. Its gastric cavity can serve as a simple model of microbial-animal digestive associations, yet poorly characterized. Using state-of-the-art metagenomic population binning and catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH), we show that only four novel clonal phylotypes were consistently associated with multiple jellyfish adults. Two affiliated close to Spiroplasma and Mycoplasma genera, one to chlamydial 'Candidatus Syngnamydia', and one to bacteroidetal Tenacibaculum, and were at least one order of magnitude more abundant than any other bacteria detected. Metabolic modelling predicted an aerobic heterotrophic lifestyle for the chlamydia, which were found intracellularly in Onychodromopsis-like ciliates. The Spiroplasma-like organism was predicted to be an anaerobic fermenter associated to some jellyfish cells, whereas the Tenacibaculum-like as free-living aerobic heterotroph, densely colonizing the mesogleal axis inside the gastric filaments. The association between the jellyfish and its reduced microbiome was close and temporally stable, and possibly related to food digestion and protection from pathogens. Based on the genomic and microscopic data, we propose three candidate taxa: 'Candidatus Syngnamydia medusae', 'Candidatus Medusoplasma mediterranei' and 'Candidatus Tenacibaculum medusae'.
Assuntos
Chlamydia/classificação , Mycoplasma/classificação , Cifozoários/microbiologia , Spiroplasma/classificação , Tenacibaculum/classificação , Animais , Biodiversidade , Chlamydia/genética , Chlamydia/isolamento & purificação , Feminino , Microbioma Gastrointestinal , Hibridização in Situ Fluorescente , Masculino , Mar Mediterrâneo , Mycoplasma/genética , Mycoplasma/isolamento & purificação , RNA Ribossômico 16S/genética , Spiroplasma/genética , Spiroplasma/isolamento & purificação , Tenacibaculum/genética , Tenacibaculum/isolamento & purificaçãoRESUMO
For maternally transmitted microbes, a female-biased host sex ratio is of reproductive advantage. Here we found a strong female bias in a field population of the green lacewing, Mallada desjardinsi (Insecta; Neuroptera). This bias was attributed to the predominance of individuals harboring a maternally inherited male-killing bacterium that was phylogenetically closely related to the plant-pathogenic Spiroplasma phoeniceum and Spiroplasma kunkelii. Among 35 laboratory-reared broods produced by wild-caught females, 21 broods (60%)-all infected with Spiroplasma-consisted of only females (940 individuals). Among 14 broods consisting of both males and females (516 and 635 individuals, respectively), 4 broods were doubly infected with Spiroplasma and Rickettsia, 6 broods were singly infected with Rickettsia, and 3 broods were uninfected (remaining one brood was unknown). Mortality during embryonic and larval development was prominent in all-female broods but not in normal sex ratio broods. Following antibiotic treatment on all-female broods, mortality was significantly reduced and the sex ratio was restored to 1:1. Strong expression and high prevalence of this male-killer is remarkable considering its low density (~10-5-10-4 cells per host mitochondrial gene copy based on quantitative PCR). In addition, a bacterium closely related to Rickettsia bellii was present in 25 of 34 broods (73.5%), irrespective of the sex ratio, with the infection density comparable to other cases of endosymbiosis (~10-2-10-1 cells per mitochondrial gene copy). Higher density of Rickettsia than Spiroplasma was also demonstrated by electron microscopy which visualized both Spiroplasma-like cells and Rickettsia-like cells inside and outside the ovarian cells.
Assuntos
Insetos/microbiologia , Rickettsia/fisiologia , Spiroplasma/fisiologia , Animais , Antibacterianos/farmacologia , DNA Bacteriano/química , DNA Bacteriano/genética , Feminino , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Larva/efeitos dos fármacos , Larva/microbiologia , Masculino , Microscopia Eletrônica de Transmissão , Ovário/microbiologia , Ovário/ultraestrutura , Filogenia , RNA Ribossômico 16S/genética , Rickettsia/classificação , Rickettsia/genética , Análise de Sequência de DNA , Fatores Sexuais , Razão de Masculinidade , Spiroplasma/classificação , Spiroplasma/genética , Simbiose , Tetraciclina/farmacologiaRESUMO
Reproductive endosymbionts have been shown to have wide-ranging effects on many aspects of their hosts' biology. A first step to understanding how these endosymbionts interact with their hosts is to determine their incidences. Here, we screened for four reproductive endosymbionts (Wolbachia, Cardinium, Spiroplasma and Rickettsia) in 28 populations of spider mites (Acari: Tetranychidae) representing 12 species. Each of the four endosymbionts were identified in at least some of the tested specimens, and their infection patterns showed variations at the species-level and population-level, suggesting their distributions can be correlated with both the phylogeny and ecology of the hosts. Co-infections of unrelated bacteria, especially double infections of Wolbachia and Cardinium within the same individuals were common. Spiroplasma and Rickettsia infections were specific to particular host species, respectively. Further, the evolutionary histories of these endosymbionts were inferred by comparing the phylogenies of them and their hosts. These findings can help to clarify the interactions between endosymbionts and arthropods.
Assuntos
Bacteroidetes/fisiologia , Rickettsia/fisiologia , Spiroplasma/fisiologia , Tetranychidae/microbiologia , Wolbachia/fisiologia , Animais , Bacteroidetes/classificação , Bacteroidetes/genética , Evolução Biológica , Coinfecção , Complexo IV da Cadeia de Transporte de Elétrons/química , Complexo IV da Cadeia de Transporte de Elétrons/genética , Especificidade de Hospedeiro , Filogenia , RNA Ribossômico 18S/química , RNA Ribossômico 18S/genética , RNA Ribossômico 28S/química , RNA Ribossômico 28S/genética , Reprodução , Rickettsia/classificação , Rickettsia/genética , Análise de Sequência de DNA , Spiroplasma/classificação , Spiroplasma/genética , Simbiose , Tetranychidae/classificação , Tetranychidae/genética , Wolbachia/classificação , Wolbachia/genéticaRESUMO
The ectoparasitic mite Varroa destructor is a major pest of the honeybee Apis mellifera. In a previous study, bacteria were found in the guts of mites collected from winter beehive debris and were identified using Sanger sequencing of their 16S rRNA genes. In this study, community comparison and diversity analyses were performed to examine the microbiota of honeybees and mites at the population level. The microbiota of the mites and honeybees in 26 colonies in seven apiaries in Czechia was studied. Between 10 and 50 Varroa females were collected from the bottom board, and 10 worker bees were removed from the peripheral comb of the same beehive. Both bees and mites were surface sterilized. Analysis of the 16S rRNA gene libraries revealed significant differences in the Varroa and honeybee microbiota. The Varroa microbiota was less diverse than was the honeybee microbiota, and the relative abundances of bacterial taxa in the mite and bee microbiota differed. The Varroa mites, but not the honeybees, were found to be inhabited by Diplorickettsia. The relative abundance of Arsenophonus, Morganella, Spiroplasma, Enterococcus, and Pseudomonas was higher in Varroa than in honeybees, and the Diplorickettsia symbiont detected in this study is specific to Varroa mites. The results demonstrated that there are shared bacteria between Varroa and honeybee populations but that these bacteria occur in different relative proportions in the honeybee and mite bacteriomes. These results support the suggestion of bacterial transfer via mites, although only some of the transferred bacteria may be harmful.
Assuntos
Abelhas/microbiologia , Microbiota , Spiroplasma/classificação , Varroidae/microbiologia , Animais , Abelhas/parasitologia , Biodiversidade , DNA Bacteriano/genética , Feminino , Masculino , RNA Ribossômico 16S/genética , Estações do Ano , Análise de Sequência de DNA , Spiroplasma/isolamento & purificação , SimbioseRESUMO
Drechmeria coniospora is an obligate fungal pathogen that infects nematodes via the adhesion of specialized spores to the host cuticle. D. coniospora is frequently found associated with Caenorhabditis elegans in environmental samples. It is used in the study of the nematode's response to fungal infection. Full understanding of this bi-partite interaction requires knowledge of the pathogen's genome, analysis of its gene expression program and a capacity for genetic engineering. The acquisition of all three is reported here. A phylogenetic analysis placed D. coniospora close to the truffle parasite Tolypocladium ophioglossoides, and Hirsutella minnesotensis, another nematophagous fungus. Ascomycete nematopathogenicity is polyphyletic; D. coniospora represents a branch that has not been molecularly characterized. A detailed in silico functional analysis, comparing D. coniospora to 11 fungal species, revealed genes and gene families potentially involved in virulence and showed it to be a highly specialized pathogen. A targeted comparison with nematophagous fungi highlighted D. coniospora-specific genes and a core set of genes associated with nematode parasitism. A comparative gene expression analysis of samples from fungal spores and mycelia, and infected C. elegans, gave a molecular view of the different stages of the D. coniospora lifecycle. Transformation of D. coniospora allowed targeted gene knock-out and the production of fungus that expresses fluorescent reporter genes. It also permitted the initial characterisation of a potential fungal counter-defensive strategy, involving interference with a host antimicrobial mechanism. This high-quality annotated genome for D. coniospora gives insights into the evolution and virulence of nematode-destroying fungi. Coupled with genetic transformation, it opens the way for molecular dissection of D. coniospora physiology, and will allow both sides of the interaction between D. coniospora and C. elegans, as well as the evolutionary arms race that exists between pathogen and host, to be studied.
Assuntos
Caenorhabditis elegans/microbiologia , Micoses/microbiologia , Filogenia , Spiroplasma/genética , Animais , Ascomicetos/genética , Ascomicetos/patogenicidade , Caenorhabditis elegans/parasitologia , Hibridização Genômica Comparativa , Hypocreales/classificação , Hypocreales/genética , Micoses/parasitologia , Spiroplasma/classificação , Spiroplasma/patogenicidade , Esporos Fúngicos/classificação , Esporos Fúngicos/genética , Esporos Fúngicos/patogenicidade , Virulência/genéticaRESUMO
Animals and plants are increasingly threatened by emerging fungal and oomycete diseases. Amongst oomycetes, Saprolegnia species cause population declines in aquatic animals, especially fish and amphibians, resulting in significant perturbation in biodiversity, ecological balance and food security. Due to the prohibition of several chemical control agents, novel sustainable measures are required to control Saprolegnia infections in aquaculture. Previously, fungal community analysis by terminal restriction fragment length polymorphism (T-RFLP) revealed that the Ascomycota, specifically the genus Microdochium, was an abundant fungal phylum associated with salmon eggs from a commercial fish farm. Here, phylogenetic analyses showed that most fungal isolates obtained from salmon eggs were closely related to Microdochium lycopodinum/Microdochium phragmitis and Trichoderma viride species. Phylogenetic and quantitative PCR analyses showed both a quantitative and qualitative difference in Trichoderma population between diseased and healthy salmon eggs, which was not the case for the Microdochium population. In vitro antagonistic activity of the fungi against Saprolegnia diclina was isolate-dependent; for most Trichoderma isolates, the typical mycoparasitic coiling around and/or formation of papilla-like structures on S. diclina hyphae were observed. These results suggest that among the fungal community associated with salmon eggs, Trichoderma species may play a role in Saprolegnia suppression in aquaculture.
Assuntos
Antibiose , Doenças dos Peixes/terapia , Infecções/terapia , Saprolegnia/microbiologia , Spiroplasma/crescimento & desenvolvimento , Trichoderma/crescimento & desenvolvimento , Animais , Aquicultura , Biodiversidade , Agentes de Controle Biológico , Doenças dos Peixes/parasitologia , Infecções/parasitologia , Filogenia , Salmão/microbiologia , Salmão/parasitologia , Saprolegnia/crescimento & desenvolvimento , Saprolegnia/patogenicidade , Spiroplasma/classificação , Spiroplasma/genética , Trichoderma/classificação , Trichoderma/genética , Zigoto/microbiologia , Zigoto/parasitologiaRESUMO
Bacteria associated with the tick Ixodes ricinus were assessed in specimens unattached or attached to the skin of cats, dogs and humans, collected in the Czech Republic. The bacteria were detected by PCR in 97 of 142 pooled samples including 204 ticks, i.e. 1-7 ticks per sample, collected at the same time from one host. A fragment of the bacterial 16S rRNA gene was amplified, cloned and sequenced from 32 randomly selected samples. The most frequent sequences were those related to Candidatus Midichloria midichlori (71% of cloned sequences), followed by Diplorickettsia (13%), Spiroplasma (3%), Rickettsia (3%), Pasteurella (3%), Morganella (3%), Pseudomonas (2%), Bacillus (1%), Methylobacterium (1%) and Phyllobacterium (1%). The phylogenetic analysis of Spiroplasma 16S rRNA gene sequences showed two groups related to Spiroplasma eriocheiris and Spiroplasma melliferum, respectively. Using group-specific primers, the following potentially pathogenic bacteria were detected: Borellia (in 20% of the 142 samples), Rickettsia (12%), Spiroplasma (5%), Diplorickettsia (5%) and Anaplasma (2%). In total, 68% of I. ricinus samples (97/142) contained detectable bacteria and 13% contained two or more putative pathogenic groups. The prevalence of tick-borne bacteria was similar to the observations in other European countries.
Assuntos
Bactérias/genética , Ixodes/microbiologia , Animais , Bactérias/classificação , Gatos/microbiologia , Gatos/parasitologia , Cidades , República Tcheca , DNA Bacteriano/genética , Cães/microbiologia , Cães/parasitologia , Feminino , Humanos , Ixodes/crescimento & desenvolvimento , Masculino , Dados de Sequência Molecular , Ninfa/microbiologia , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Spiroplasma/classificação , Spiroplasma/genéticaRESUMO
Spiroplasma is a genus of wall-less, low-GC, small Gram-positive bacteria of the internal contractile cytoskeleton, with helical morphology and motility. The genus is classified within the class Mollicutes. Spiroplasma / host interactions can be classified as commensal, pathogenic or mutualist. The majority of spiroplasmas are found to be commensals of insects, arachnids, crustaceans or plants, whereas a small number of species are pathogens of plants, insects, and crustaceans. Insects are particularly rich sources of spiroplasmas. The bacteria are common in haematophagous arthropods: deerflies, horseflies, mosquitoes, and in ticks, where they may occur abundantly in salivary glands. The ability of spiroplasmas to propagate in rodents was experimentally proven, and Spiroplasma infections have been reported recently in humans. Some authors have purported an etiological role of Spiroplasma in causing transmissible spongiform encephalopathies (TSEs), but convincing proof is lacking. The possibility for humans and other vertebrates to be infected with Spiroplasma spp. in natural conditions is largely unknown, as well as the possibility of the transmission of these bacteria by ticks and haematophagous insects. Nevertheless, in the light of new data, such possibilities cannot be excluded.
Assuntos
Doenças Transmissíveis Emergentes/microbiologia , Dípteros/microbiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Spiroplasma/fisiologia , Carrapatos/microbiologia , Animais , Doenças Transmissíveis Emergentes/transmissão , Infecções por Bactérias Gram-Negativas/transmissão , Humanos , Spiroplasma/classificação , VertebradosRESUMO
Horizontal gene transfer (HGT) is an important mechanism that contributed to biological diversity, particularly in bacteria. Through acquisition of novel genes, the recipient cell may change its ecological preference and the process could promote speciation. In this study, we determined the complete genome sequence of two Spiroplasma species for comparative analyses and inferred the putative gene gains and losses. Although most Spiroplasma species are symbionts of terrestrial insects, Spiroplasma eriocheiris has evolved to be a lethal pathogen of freshwater crustaceans. We found that approximately 7% of the genes in this genome may have originated from HGT and these genes expanded the metabolic capacity of this organism. Through comparison with the closely related Spiroplasma atrichopogonis, as well as other more divergent lineages, our results indicated that these HGT events could be traced back to the most recent common ancestor of these two species. However, most of these horizontally acquired genes have been pseudogenized in S. atrichopogonis, suggesting that they did not contribute to the fitness of this lineage that maintained the association with terrestrial insects. Thus, accumulation of small deletions that disrupted these foreign genes was not countered by natural selection. On the other hand, the long-term survival of these horizontally acquired genes in the S. eriocheiris genome hinted that they might play a role in the ecological shift of this species. Finally, the implications of these findings and the conflicts among gene content, 16S rRNA gene sequencing, and serological typing, are discussed in light of defining bacterial species.