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1.
Artigo em Inglês | MEDLINE | ID: mdl-37822789

RESUMO

The genus Leptoconops Skuse (Diptera: Ceratopogonidae) are blood-sucking midges known to pester humans and domestic animals. In certain Mediterranean areas, midges occur in large numbers during summer and limit the use of recreational areas, also raising serious health and social concerns. Despite such impact, the diversity and distribution of Leptoconops in Maremma Regional Park (Tuscany Region, Italy), a heavily infested area, is not well known, and neither molecular nor detailed morphological studies exist. We sampled adult midge females in six areas and used high-resolution digital stereomicroscopy and scanning electron microscopy to identify species and investigate the morphology of structures involved in host searching/recognition (antennae and maxillary palps) and host attack (mouthparts). We also performed energy-dispersive X-ray spectroscopy to characterize the elemental composition of mouthparts. Finally, the cytochrome c oxidase subunit 1 (cox1) gene was amplified and sequenced, to confirm species identification of collected specimens. We identified two species: Leptoconops (L.) irritans Noé and Leptoconops (L.) noei Clastrier & Coluzzi, with the former being more frequently sampled than the latter and closer to sea coast and rivers. The antennal segments appeared slightly more globular in L. noei than in L. irritans. Five types of trichoid, basiconic and chaetic sensilla were found on the antennae, with some differences between the two species. Mouthparts had the labellum visibly larger in L. noei compared with L. irritans. The maxillary palps possessed a pit filled with bulb-shaped sensilla, which appeared denser in L. noei than in L. irritans. Mouthpart cuticle included Calcium (Ca) and Aluminum (Al) at small but significant concentrations (0.3-1.0%) in both species. Our results suggest that the limited but appreciable differences in sensory system between the studied species of Leptoconops and other Ceratopogonidae may reflect different host or habitat preferences, a scenario potentially suggested also by preliminarily data on their distribution in the studied area. The presence of Ca and Al in the cuticle of mouthparts may help host skin drilling during bite activity. Finally, the gene sequences obtained in this study provide a first reference for future investigations on the taxonomy and dispersal patterns of Leptoconops spp. in the Mediterranean area.

2.
Parasit Vectors ; 16(1): 35, 2023 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-36703216

RESUMO

Parasites of the genus Leishmania are unusual unicellular microorganisms in that they are characterized by the capability to subvert in their favor the immune response of mammalian phagocytes, including dendritic cells. Thus, in overt leishmaniasis, dendritic cells and macrophages are converted into a niche for Leishmania spp. in which the parasite, rather than being inactivated and disassembled, survives and replicates. In addition, Leishmania parasites hitchhike onto phagocytic cells, exploiting them as a mode of transport to lymphoid tissues where other phagocytic cells are potentially amenable to parasite colonization. This propensity of Leishmania spp. to target dendritic cells has led some researchers to consider the possibility that the non-pathogenic, reptile-associated Leishmania tarentolae could be exploited as a vaccine platform and vehicle for the production of antigens from different viruses and for the delivery of the antigens to dendritic cells and lymph nodes. In addition, as L. tarentolae can also be regarded as a surrogate of pathogenic Leishmania parasites, this parasite of reptiles could possibly be developed into a vaccine against human and canine leishmaniases, exploiting its immunological cross-reactivity with other Leishmania species, or, after its engineering, for the expression of antigens from pathogenic species. In this article we review published studies on the use of L. tarentolae as a vaccine platform and vehicle, mainly in the areas of leishmaniases and viral infections. In addition, a short summary of available knowledge on the biology of L. tarentolae is presented, together with information on the use of this microorganism as a micro-factory to produce antigens suitable for the serodiagnosis of viral and parasitic infections.


Assuntos
Leishmania , Leishmaniose , Parasitos , Vacinas , Viroses , Animais , Cães , Humanos , Leishmaniose/prevenção & controle , Leishmaniose/parasitologia , Células Dendríticas , Mamíferos
3.
Plants (Basel) ; 11(24)2022 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-36559686

RESUMO

Grapevine cultivation, such as the whole horticulture, is currently challenged by several factors, among which the extreme weather events occurring under the climate change scenario are the most relevant. Within this context, the present study aims at characterizing at the berry level the physiological response of Vitis vinifera cv. Sauvignon Blanc to sequential stresses simulated under a semi-controlled environment: flooding at bud-break followed by multiple summer stress (drought plus heatwave) occurring at pre-vèraison. Transcriptomic and metabolomic assessments were performed through RNASeq and NMR, respectively. A comprehensive hormone profiling was also carried out. Results pointed out a different response to the heatwave in the two situations. Flooding caused a developmental advance, determining a different physiological background in the berry, thus affecting its response to the summer stress at both transcriptional levels, with the upregulation of genes involved in oxidative stress responses, and metabolic level, with the increase in osmoprotectants, such as proline and other amino acids. In conclusion, sequential stress, including a flooding event at bud-break followed by a summer heatwave, may impact phenological development and berry ripening, with possible consequences on berry and wine quality. A berry physiological model is presented that may support the development of sustainable vineyard management solutions to improve the water use efficiency and adaptation capacity of actual viticultural systems to future scenarios.

4.
Microorganisms ; 10(6)2022 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-35744681

RESUMO

Staphylococcus epidermidis is an opportunistic pathogen and a frequent cause of nosocomial infections. In this work, we show that, among 51 S. epidermidis isolates from an Italian hospital, only a minority displayed biofilm formation, regardless of their isolation source (peripheral blood, catheter, or skin wounds); however, among the biofilm-producing isolates, those from catheters were the most efficient in biofilm formation. Interestingly, most isolates including strong biofilm producers displayed production levels of PIA (polysaccharide intercellular adhesin), the main S. epidermidis extracellular polysaccharide, similar to reference S. epidermidis strains classified as non-biofilm formers, and much lower than those classified as intermediate or high biofilm formers, possibly suggesting that high levels of PIA production do not confer a particular advantage for clinical isolates. Finally, while for the reference S. epidermidis strains the biofilm production clearly correlated with the decreased sensitivity to antibiotics, in particular, protein synthesis inhibitors, in our clinical isolates, such positive correlation was limited to tetracycline. In contrast, we observed an inverse correlation between biofilm formation and the minimal inhibitory concentrations for levofloxacin and teicoplanin. In addition, in growth conditions favoring PIA production, the biofilm-forming isolates showed increased sensitivity to daptomycin, clindamycin, and erythromycin, with increased tolerance to the trimethoprim/sulfamethoxazole association. The lack of direct correlation between the biofilm production and increased tolerance to antibiotics in S. epidermidis isolates from a clinical setting would suggest, at least for some antimicrobials, the possible existence of a trade-off between the production of biofilm determinants and antibiotic resistance.

5.
PLoS Biol ; 20(2): e3001528, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-35192605

RESUMO

Bacteria are powerful models for understanding how cells divide and accomplish global regulatory programs. In Caulobacter crescentus, a cascade of essential master regulators supervises the correct and sequential activation of DNA replication, cell division, and development of different cell types. Among them, the response regulator CtrA plays a crucial role coordinating all those functions. Here, for the first time, we describe the role of a novel factor named CcnA (cell cycle noncoding RNA A), a cell cycle-regulated noncoding RNA (ncRNA) located at the origin of replication, presumably activated by CtrA, and responsible for the accumulation of CtrA itself. In addition, CcnA may be also involved in the inhibition of translation of the S-phase regulator, GcrA, by interacting with its 5' untranslated region (5' UTR). Performing in vitro experiments and mutagenesis, we propose a mechanism of action of CcnA based on liberation (ctrA) or sequestration (gcrA) of their ribosome-binding site (RBS). Finally, its role may be conserved in other alphaproteobacterial species, such as Sinorhizobium meliloti, representing indeed a potentially conserved process modulating cell cycle in Caulobacterales and Rhizobiales.


Assuntos
Caulobacter crescentus , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Caulobacter crescentus/genética , Caulobacter crescentus/metabolismo , Ciclo Celular/genética , Proteínas de Ligação a DNA/metabolismo , Regulação Bacteriana da Expressão Gênica/genética , Regiões Promotoras Genéticas , RNA não Traduzido/genética , Fatores de Transcrição/metabolismo
6.
Parasit Vectors ; 14(1): 534, 2021 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-34649599

RESUMO

BACKGROUND: Aedes koreicus is a mosquito species characterized by marked anthropophilic behavior, and a potential vector of nematodes and viruses. It is native to East Asia, but its presence has recently been reported in many regions of Europe. In Italy, these mosquitoes had been detected in the northeast since 2011 and are now spreading towards the southwest of the country. METHODS: In 2020, during a surveillance program for invasive mosquito species in the district of Bergamo (Lombardy Region, Italy), about 6000 mosquito larvae were collected. Emerged adults were assigned to mosquito species according to morphological analyses, followed by amplification and sequencing of genetic markers (COI, ND4, ITS2 and D2). RESULTS: According to the morphological and genetic data, about 50 individuals belonged to the species Ae. koreicus. CONCLUSION: We report the presence of Ae. koreicus in the district of Bergamo, which confirms the spread of this species in the north of Italy and raises concerns about its possible role as a vector of diseases in the Alpine area.


Assuntos
Aedes/fisiologia , Espécies Introduzidas , Mosquitos Vetores/fisiologia , Aedes/anatomia & histologia , Aedes/classificação , Aedes/genética , Distribuição Animal , Animais , Feminino , Itália , Larva/fisiologia , Masculino , Mosquitos Vetores/anatomia & histologia , Mosquitos Vetores/classificação , Mosquitos Vetores/genética
7.
Int J Mol Sci ; 21(18)2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32942778

RESUMO

3D cell cultures are becoming more and more important in the field of regenerative medicine due to their ability to mimic the cellular physiological microenvironment. Among the different types of 3D scaffolds, we focus on the Nichoid, a miniaturized scaffold with a structure inspired by the natural staminal niche. The Nichoid can activate cellular responses simply by subjecting the cells to mechanical stimuli. This kind of influence results in different cellular morphology and organization, but the molecular bases of these changes remain largely unknown. Through RNA-Seq approach on murine neural precursors stem cells expanded inside the Nichoid, we investigated the deregulated genes and pathways showing that the Nichoid causes alteration in genes strongly connected to mechanobiological functions. Moreover, we fully dissected this mechanism highlighting how the changes start at a membrane level, with subsequent alterations in the cytoskeleton, signaling pathways, and metabolism, all leading to a final alteration in gene expression. The results shown here demonstrate that the Nichoid influences the biological and genetic response of stem cells thorough specific alterations of cellular signaling. The characterization of these pathways elucidates the role of mechanical manipulation on stem cells, with possible implications in regenerative medicine applications.


Assuntos
Mecanotransdução Celular/genética , Células-Tronco Neurais/metabolismo , Transcriptoma/genética , Animais , Técnicas de Cultura de Células , Células Cultivadas , Citoesqueleto/genética , Expressão Gênica/genética , Camundongos , Camundongos Endogâmicos C57BL , Medicina Regenerativa/métodos , Transdução de Sinais/genética , Nicho de Células-Tronco/genética , Alicerces Teciduais/química
8.
Int J Mol Sci ; 21(14)2020 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-32668813

RESUMO

Modulation of nutrient digestion and absorption is one of the post-ingestion mechanisms that guarantees the best exploitation of food resources, even when they are nutritionally poor or unbalanced, and plays a pivotal role in generalist feeders, which experience an extreme variability in diet composition. Among insects, the larvae of black soldier fly (BSF), Hermetia illucens, can grow on a wide range of feeding substrates with different nutrient content, suggesting that they can set in motion post-ingestion processes to match their nutritional requirements. In the present study we address this issue by investigating how the BSF larval midgut adapts to diets with different nutrient content. Two rearing substrates were compared: a nutritionally balanced diet for dipteran larvae and a nutritionally poor diet that mimics fruit and vegetable waste. Our data show that larval growth performance is only moderately affected by the nutritionally poor diet, while differences in the activity of digestive enzymes, midgut cell morphology, and accumulation of long-term storage molecules can be observed, indicating that diet-dependent adaptation processes in the midgut ensure the exploitation of poor substrates. Midgut transcriptome analysis of larvae reared on the two substrates showed that genes with important functions in digestion and absorption are differentially expressed, confirming the adaptability of this organ.


Assuntos
Dieta , Dípteros/fisiologia , Adaptação Fisiológica , Ração Animal/análise , Animais , Peso Corporal , Carboidratos da Dieta/análise , Carboidratos da Dieta/farmacocinética , Proteínas Alimentares/análise , Proteínas Alimentares/farmacocinética , Dípteros/genética , Dípteros/crescimento & desenvolvimento , Frutas , Regulação da Expressão Gênica , Ontologia Genética , Concentração de Íons de Hidrogênio , Absorção Intestinal , Mucosa Intestinal/metabolismo , Intestinos/fisiologia , Larva , Nutrientes/análise , Nutrientes/farmacocinética , Pupa , RNA-Seq , Transcriptoma , Verduras
9.
Genes (Basel) ; 11(3)2020 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-32121150

RESUMO

Mgaloblishvili, a Vitis vinifera cultivar, exhibits unique resistance traits against Plasmopara viticola, the downy mildew agent. This offers the unique opportunity of exploring the molecular responses in compatible and incompatible plant-pathogen interaction. In this study, whole transcriptomes of Mgaloblishvili, Pinot noir (a V. vinifera susceptible cultivar), and Bianca (a resistant hybrid) leaves, inoculated and non-inoculated with the pathogen, were used to identify P. viticola effector-encoding genes and plant susceptibility/resistance genes. Multiple effector-encoding genes were identified in P. viticola transcriptome, with remarkable expression differences in relation to the inoculated grapevine cultivar. Intriguingly, five apoplastic effectors specifically associated with resistance in V. vinifera. Gene coexpression network analysis identified specific modules and metabolic changes occurring during infection in the three grapevine cultivars. Analysis of these data allowed, for the first time, the detection in V. vinifera of a putative P. viticola susceptibility gene, encoding a LOB domain-containing protein. Finally, the de novo assembly of Mgaloblishvili, Pinot noir, and Bianca transcriptomes and their comparison highlighted novel candidate genes that might be at the basis of the resistant phenotype. These results open the way to functional analysis studies and to new perspectives in molecular breeding of grapevine for resistance to P. viticola.


Assuntos
Resistência à Doença/genética , Doenças das Plantas/genética , Transcriptoma/genética , Vitis/genética , Regulação da Expressão Gênica de Plantas/genética , Interações Hospedeiro-Patógeno/genética , Oomicetos/genética , Oomicetos/patogenicidade , Doenças das Plantas/microbiologia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/microbiologia , Análise de Sequência de RNA , Vitis/crescimento & desenvolvimento , Vitis/microbiologia
10.
Front Genet ; 10: 853, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31608107

RESUMO

In recent years, the advent of NGS technology has made genome sequencing much cheaper than in the past; the high parallelization capability and the possibility to sequence more than one organism at once have opened the door to processing whole symbiotic consortia. However, this approach needs the development of specific bioinformatics tools able to analyze these data. In this work, we describe SeqDex, a tool that starts from a preliminary assembly obtained from sequencing a mixture of DNA from different organisms, to identify the contigs coming from one organism of interest. SeqDex is a fully automated machine learning-based tool exploiting partial taxonomic affiliations and compositional analysis to predict the taxonomic affiliations of contigs in an assembly. In literature, there are few methods able to deconvolve host-symbiont datasets, and most of them heavily rely on user curation and are therefore time consuming. The problem has strong similarities with metagenomic studies, where mixed samples are sequenced and the bioinformatics challenge is trying to separate contigs on the basis of their source organism; however, in symbiotic systems, additional information can be exploited to improve the output. To assess the ability of SeqDex to deconvolve host-symbiont datasets, we compared it to state-of-the-art methods for metagenomic binning and for host-symbiont deconvolution on three study cases. The results point out the good performances of the presented tool that, in addition to the ease of use and customization potential, make SeqDex a useful tool for rapid identification of endosymbiont sequences.

11.
Front Plant Sci ; 10: 339, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30972087

RESUMO

Studies on model plants have shown that temporary soil flooding exposes roots to a significant hypoxic stress resulting in metabolic re-programming, accumulation of toxic metabolites and hormonal imbalance. To date, physiological and transcriptional responses to flooding in grapevine are poorly characterized. To fill this gap, we aimed to gain insights into the transcriptional and metabolic changes induced by flooding on grapevine roots (K5BB rootstocks), on which cv Sauvignon blanc (Vitis vinifera L.) plants were grafted. A preliminary experiment under hydroponic conditions enabled the identification of transiently and steadily regulated hypoxia-responsive marker genes and drafting a model for response to oxygen deprivation in grapevine roots. Afterward, over two consecutive vegetative seasons, flooding was imposed to potted vines during the late dormancy period, to mimick the most frequent waterlogging events occurring in the field. Untargeted transcriptomic and metabolic profiling approaches were applied to investigate early responses of grapevine roots during exposure to hypoxia and subsequent recovery after stress removal. The initial hypoxic response was marked by a significant increase of the hypoxia-inducible metabolites ethanol, GABA, succinic acid and alanine which remained high also 1 week after recovery from flooding with the exception of ethanol that leveled off. Transcriptomic data supported the metabolic changes by indicating a substantial rearrangement of primary metabolic pathways through enhancement of the glycolytic and fermentative enzymes and of a subset of enzymes involved in the TCA cycle. GO and KEGG pathway analyses of differentially expressed genes showed a general down-regulation of brassinosteroid, auxin and gibberellin biosynthesis in waterlogged plants, suggesting a general inhibition of root growth and lateral expansion. During recovery, transcriptional activation of gibberellin biosynthetic genes and down-regulation of the metabolic ones may support a role for gibberellins in signaling grapevine rootstocks waterlogging metabolic and hormonal changes to the above ground plant. The significant internode elongation measured upon budbreak during recovery in plants that had experienced flooding supported this hypothesis. Overall integration of these data enabled us to draft a first comprehensive view of the molecular and metabolic pathways involved in grapevine's root responses highlighting a deep metabolic and transcriptomic reprogramming during and after exposure to waterlogging.

12.
Microb Drug Resist ; 25(5): 787-790, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30589602

RESUMO

In the past years, the Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae sequence type 258 (ST258) became an important worldwide spread nosocomial pathogen. Recent evidence shows that the global epidemiology is changing, with the rise of new lineages. In this study we report the microbiological and genomic features of two VIM-1-producing K. pneumoniae isolates belonging to the emerging ST307. Two extensively drug-resistant K. pneumoniae strains, collected between May and June 2017, were confirmed as blaVIM positive by GeneXpert system. The whole-genome sequencing revealed that both KpV_S_1 and KpV_S_2 isolates harbored blaVIM-1 and blaCTX-M-15 genes, besides qnrS1 and qnrB1, strB, mphA, tetR, and tetA determinants. KpV_S_1 and KpV_S_2 isolates belonged to ST661 and ST307, respectively. Both STs have been recently reported as responsible of outbreaks in several European countries. The detection of blaVIM-1 gene in nonpredominant K. pneumoniae clones in a hospital setting should alert on the changing of the epidemiological situation in Italy, usually endemic reservoir of KPC enzyme.


Assuntos
Antibacterianos/farmacologia , Infecção Hospitalar/epidemiologia , Farmacorresistência Bacteriana Múltipla/genética , Genes Bacterianos , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/genética , Aminoglicosídeos/farmacologia , Carbapenêmicos/farmacologia , Cefalosporinas/farmacologia , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , Fluoroquinolonas/farmacologia , Expressão Gênica , Hospitais , Humanos , Itália/epidemiologia , Infecções por Klebsiella/tratamento farmacológico , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/classificação , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/isolamento & purificação , Macrolídeos/farmacologia , Testes de Sensibilidade Microbiana , Penicilinas/farmacologia , Tetraciclinas/farmacologia , Sequenciamento Completo do Genoma , beta-Lactamases/genética
13.
Planta ; 248(5): 1143-1157, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30066220

RESUMO

MAIN CONCLUSION: A coordinated regulation of different metabolic pathways was highlighted leading to the accumulation of important compounds that may contribute to the final quality of strawberry fruit. Strawberry fruit development and ripening involve complex physiological and biochemical changes, ranging from sugar accumulation to the production of important volatiles compounds that contribute to the final fruit flavor. To better understand the mechanisms controlling fruit growth and ripening in cultivated strawberry (Fragaria × ananassa), we applied a molecular approach combining suppression subtractive hybridization and next generation sequencing to identify genes regulating developmental stages going from fruit set to full ripening. The results clearly indicated coordinated regulation of several metabolic processes such as the biosynthesis of flavonoid, phenylpropanoid and branched-chain amino acids, together with glycerolipid metabolism and pentose and glucuronate interconversion. In particular, genes belonging to the flavonoid pathway were activated in two distinct phases, the first one at the very early stages of fruit development and the second during ripening. The combination of expression analysis with metabolomic data revealed that the functional meaning of these two inductions is different, as during the early stages gene activation of flavonoid pathway leads to the production of proanthocyanidins and ellagic acid-derived tannins, while during ripening anthocyanins are the main product of flavonoid pathway activation. Moreover, the subtractive approach allowed the identification of different members of the same gene family coding for the same or very similar enzymes that in some cases showed opposite regulation during strawberry fruit development. Such regulation is an important trait that can help to understand how plants specifically channel metabolic intermediates towards separate branches of a biosynthetic pathway or use different isoforms of the same enzyme in different organs or developmental stages.


Assuntos
Fragaria/metabolismo , Frutas/metabolismo , Flavonoides/metabolismo , Fragaria/genética , Fragaria/crescimento & desenvolvimento , Frutas/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas/genética , Redes e Vias Metabólicas , Metabolômica , Análise de Sequência de DNA , Técnicas de Hibridização Subtrativa , Transcriptoma
14.
New Microbiol ; 41(3): 230-231, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-30028475

RESUMO

We describe two multi drug-resistant (MDR) carbapenemase-producing Escherichia coli clinical isolates from an acute hospital in Milan. Both strains, isolated from a surgical wound sample and a surveillance rectal swab respectively, were positive for a blaNDM-type gene by Xpert Carba-R test. The whole-genome sequence characterization disclosed several resistance determinants: blaNDM-5, blaCMY-42, blaTEM-198, rmtB, mphA. The two isolates belonged to phylogenetic group A, sequence type (ST) 1702 and serotype O89:H9. PCR-based replicon typing and conjugation assay demonstrated an IncI1 plasmid localization for both blaNDM-5 and blaCMY-42 genes. This is the first report of a ST1702 NDM-5 and CMY-42- producing E. coli clone in Italy.


Assuntos
Escherichia coli/genética , Escherichia coli/isolamento & purificação , Idoso , Antibacterianos/farmacologia , Proteínas de Bactérias , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Feminino , Regulação Bacteriana da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Hospitais , Humanos , Itália/epidemiologia , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Filogenia , Reto/microbiologia , Infecção da Ferida Cirúrgica/epidemiologia , Infecção da Ferida Cirúrgica/microbiologia , beta-Lactamases
15.
Front Genet ; 9: 164, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29899753

RESUMO

Caulobacter crescentus represents a remarkable model system to investigate global regulatory programs in bacteria. In particular, several decades of intensive study have revealed that its cell cycle is controlled by a cascade of master regulators, such as DnaA, GcrA, CcrM, and CtrA, that are responsible for the activation of functions required to progress through DNA replication, cell division and morphogenesis of polar structures (flagellum and stalk). In order to accomplish this task, several post-translational (phosphorylation and proteolysis) and transcriptional mechanisms are involved. Surprisingly, the role of non-coding RNAs (ncRNAs) in regulating the cell cycle has not been investigated. Here we describe a bioinformatic analysis that revealed that ncRNAs may well play a crucial role regulating cell cycle in C. crescentus. We used available prediction tools to understand which target genes may be regulated by ncRNAs in this bacterium. Furthermore, we predicted whether ncRNAs with a cell cycle regulated expression profile may be directly regulated by DnaA, GcrA, and CtrA, at the onset, during or end of the S-phase/swarmer cell, or if any of them has CcrM methylation sites in the promoter region. Our analysis suggests the existence of a potentially very important network of ncRNAs regulated by or regulating well-known cell cycle genes in C. crescentus. Our hypothesis is that ncRNAs are intimately connected to the known regulatory network, playing a crucial modulatory role in cell cycle progression.

16.
Gigascience ; 7(4): 1-14, 2018 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-29659812

RESUMO

Background: The genus Potentilla is closely related to that of Fragaria, the economically important strawberry genus. Potentilla micrantha is a species that does not develop berries but shares numerous morphological and ecological characteristics with Fragaria vesca. These similarities make P. micrantha an attractive choice for comparative genomics studies with F. vesca. Findings: In this study, the P. micrantha genome was sequenced and annotated, and RNA-Seq data from the different developmental stages of flowering and fruiting were used to develop a set of gene predictions. A 327 Mbp sequence and annotation of the genome of P. micrantha, spanning 2674 sequence contigs, with an N50 size of 335,712, estimated to cover 80% of the total genome size of the species was developed. The genus Potentilla has a characteristically larger genome size than Fragaria, but the recovered sequence scaffolds were remarkably collinear at the micro-syntenic level with the genome of F. vesca, its closest sequenced relative. A total of 33,602 genes were predicted, and 95.1% of bench-marking universal single-copy orthologous genes were complete within the presented sequence. Thus, we argue that the majority of the gene-rich regions of the genome have been sequenced. Conclusions: Comparisons of RNA-Seq data from the stages of floral and fruit development revealed genes differentially expressed between P. micrantha and F. vesca.The data presented are a valuable resource for future studies of berry development in Fragaria and the Rosaceae and they also shed light on the evolution of genome size and organization in this family.


Assuntos
Flores/genética , Fragaria/genética , Frutas/genética , Genoma de Planta , Potentilla/genética , Flores/crescimento & desenvolvimento , Fragaria/crescimento & desenvolvimento , Frutas/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Filogenia , Potentilla/crescimento & desenvolvimento , Análise de Sequência de RNA , Transcriptoma , Sequenciamento Completo do Genoma
17.
Microb Drug Resist ; 24(8): 1207-1209, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29473791

RESUMO

We investigated an Italian OXA-181-producing Escherichia coli clinical isolate (ECS1_14) by whole-genome sequencing. The strain coharbored blaCTX-M-15, blaCMY-2, and qnrS1 genes; it belonged to ST410(Achtman)/ST692(Pasteur) and phylogroup A. The blaOXA-181 gene was harbored on a plasmid highly similar (99% identity) to the pOXA181_EC14828 plasmid, recently reported in China.


Assuntos
Infecções por Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Escherichia coli/genética , Escherichia coli/isolamento & purificação , beta-Lactamases/genética , Idoso , Humanos , Itália , Masculino , Sequenciamento Completo do Genoma/métodos
18.
Sci Rep ; 8(1): 757, 2018 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-29335535

RESUMO

Fungicides are applied intensively to prevent downy mildew infections of grapevines (Vitis vinifera) with high impact on the environment. In order to develop alternative strategies we sequenced the genome of the oomycete pathogen Plasmopara viticola causing this disease. We show that it derives from a Phytophthora-like ancestor that switched to obligate biotrophy by losing genes involved in nitrogen metabolism and γ-Aminobutyric acid catabolism. By combining multiple omics approaches we characterized the pathosystem and identified a RxLR effector that trigger an immune response in the wild species V. riparia. This effector is an ideal marker to screen novel grape resistant varieties. Our study reveals an unprecedented bidirectional noncoding RNA-based mechanism that, in one direction might be fundamental for P. viticola to proficiently infect its host, and in the other might reduce the effects of the infection on the plant.


Assuntos
Interações Hospedeiro-Patógeno , Oomicetos/crescimento & desenvolvimento , Oomicetos/genética , Doenças das Plantas/microbiologia , Vitis/microbiologia , Inativação Gênica , Doenças das Plantas/imunologia , Análise de Sequência de DNA , Vitis/imunologia
19.
Mol Microbiol ; 107(2): 142-163, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29178391

RESUMO

Peptidoglycan is the predominant stress-bearing structure in the cell envelope of most bacteria, and also a potent stimulator of the eukaryotic immune system. Obligate intracellular bacteria replicate exclusively within the interior of living cells, an osmotically protected niche. Under these conditions peptidoglycan is not necessarily needed to maintain the integrity of the bacterial cell. Moreover, the presence of peptidoglycan puts bacteria at risk of detection and destruction by host peptidoglycan recognition factors and downstream effectors. This has resulted in a selective pressure and opportunity to reduce the levels of peptidoglycan. In this review we have analysed the occurrence of genes involved in peptidoglycan metabolism across the major obligate intracellular bacterial species. From this comparative analysis, we have identified a group of predicted 'peptidoglycan-intermediate' organisms that includes the Chlamydiae, Orientia tsutsugamushi, Wolbachia and Anaplasma marginale. This grouping is likely to reflect biological differences in their infection cycle compared with peptidoglycan-negative obligate intracellular bacteria such as Ehrlichia and Anaplasma phagocytophilum, as well as obligate intracellular bacteria with classical peptidoglycan such as Coxiella, Buchnera and members of the Rickettsia genus. The signature gene set of the peptidoglycan-intermediate group reveals insights into minimal enzymatic requirements for building a peptidoglycan-like sacculus and/or division septum.


Assuntos
Bactérias , Interações entre Hospedeiro e Microrganismos , Espaço Intracelular/microbiologia , Peptidoglicano/genética , Peptidoglicano/metabolismo , Anaplasma marginale/classificação , Anaplasma marginale/genética , Anaplasma marginale/imunologia , Anaplasma marginale/metabolismo , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/imunologia , Bactérias/metabolismo , Parede Celular/metabolismo , Chlamydia/classificação , Chlamydia/genética , Chlamydia/imunologia , Chlamydia/metabolismo , Citoplasma/metabolismo , Genoma Bacteriano/genética , Humanos , Imunidade Inata/imunologia , Orientia tsutsugamushi/classificação , Orientia tsutsugamushi/genética , Orientia tsutsugamushi/imunologia , Orientia tsutsugamushi/metabolismo , Peptidoglicano/química , Filogenia , Wolbachia/classificação , Wolbachia/genética , Wolbachia/imunologia , Wolbachia/metabolismo
20.
PLoS Genet ; 13(11): e1007103, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29161263

RESUMO

The FrzCD chemoreceptor from the gliding bacterium Myxococcus xanthus forms cytoplasmic clusters that occupy a large central region of the cell body also occupied by the nucleoid. In this work, we show that FrzCD directly binds to the nucleoid with its N-terminal positively charged tail and recruits active signaling complexes at this location. The FrzCD binding to the nucleoid occur in a DNA-sequence independent manner and leads to the formation of multiple distributed clusters that explore constrained areas. This organization might be required for cooperative interactions between clustered receptors as observed in membrane-bound chemosensory arrays.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Quimiotaxia/genética , Citoplasma/metabolismo , Myxococcus xanthus/metabolismo , Ligação Proteica , Transdução de Sinais/genética
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