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1.
Emerg Infect Dis ; 27(7): 1821-1830, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34152951

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in late 2019, and the outbreak rapidly evolved into the current coronavirus disease pandemic. SARS-CoV-2 is a respiratory virus that causes symptoms similar to those caused by influenza A and B viruses. On July 2, 2020, the US Food and Drug Administration granted emergency use authorization for in vitro diagnostic use of the Influenza SARS-CoV-2 Multiplex Assay. This assay detects influenza A virus at 102.0, influenza B virus at 102.2, and SARS-CoV-2 at 100.3 50% tissue culture or egg infectious dose, or as few as 5 RNA copies/reaction. The simultaneous detection and differentiation of these 3 major pathogens increases overall testing capacity, conserves resources, identifies co-infections, and enables efficient surveillance of influenza viruses and SARS-CoV-2.


Assuntos
COVID-19 , Vírus da Influenza A , Humanos , Vírus da Influenza A/genética , Vírus da Influenza B/genética , Reação em Cadeia da Polimerase Multiplex , Transcrição Reversa , SARS-CoV-2
2.
New Phytol ; 226(5): 1256-1262, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-31997351

RESUMO

Access to greater genomic resolution through new sequencing technologies is transforming the field of plant pathology. As scientists embrace these new methods, some overarching patterns and observations come into focus. Evolutionary genomic studies are used to determine not only the origins of pathogen lineages and geographic patterns of genetic diversity, but also to discern how natural selection structures genetic variation across the genome. With greater and greater resolution, we can now pinpoint the targets of selection on a large scale. At multiple levels, crypsis and convergent evolution are evident. Host jumps and shifts may be more pervasive than once believed, and hybridization and horizontal gene transfer (HGT) likely play important roles in the emergence of genetic novelty.


Assuntos
Genoma , Genômica , Evolução Molecular , Transferência Genética Horizontal , Hibridização Genética , Seleção Genética
3.
J Exp Bot ; 71(20): 6444-6459, 2020 10 22.
Artigo em Inglês | MEDLINE | ID: mdl-32725118

RESUMO

Recent work has provided evidence for the occurrence of N-hydroxypipecolic acid (NHP) in Arabidopsis thaliana, characterized its pathogen-inducible biosynthesis by a three-step metabolic sequence from l-lysine, and established a central role for NHP in the regulation of systemic acquired resistance. Here, we show that NHP is biosynthesized in several other plant species in response to microbial attack, generally together with its direct metabolic precursor pipecolic acid and the phenolic immune signal salicylic acid. For example, NHP accumulates locally in inoculated leaves and systemically in distant leaves of cucumber in response to Pseudomonas syringae attack, in Pseudomonas-challenged tobacco and soybean leaves, in tomato inoculated with the oomycete Phytophthora infestans, in leaves of the monocot Brachypodium distachyon infected with bacterial (Xanthomonas translucens) and fungal (Magnaporthe oryzae) pathogens, and in M. oryzae-inoculated barley. Notably, resistance assays indicate that NHP acts as a potent inducer of acquired resistance to bacterial and fungal infection in distinct monocotyledonous and dicotyledonous species. Pronounced systemic accumulation of NHP in leaf phloem sap of locally inoculated cucumber supports a function for NHP as a phloem-mobile immune signal. Our study thus generalizes the existence and function of an NHP resistance pathway in plant systemic acquired resistance.


Assuntos
Arabidopsis , Xanthomonas , Ascomicetos , Ácidos Pipecólicos , Doenças das Plantas , Folhas de Planta , Pseudomonas syringae , Ácido Salicílico
4.
BMC Evol Biol ; 19(1): 141, 2019 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-31296160

RESUMO

BACKGROUND: The LysM receptor-like kinases (LysM-RLKs) are important to both plant defense and symbiosis. Previous studies described three clades of LysM-RLKs: LysM-I/LYKs (10+ exons per gene and containing conserved kinase residues), LysM-II/LYRs (1-5 exons per gene, lacking conserved kinase residues), and LysM-III (two exons per gene, with a kinase unlike other LysM-RLK kinases and restricted to legumes). LysM-II gene products are presumably not functional as conventional receptor kinases, but several are known to operate in complexes with other LysM-RLKs. One aim of our study was to take advantage of recently mapped wild tomato transcriptomes to evaluate the evolutionary history of LysM-RLKs within and between species. The second aim was to place these results into a broader phylogenetic context by integrating them into a sequence analysis of LysM-RLKs from other functionally well-characterized model plant species. Furthermore, we sought to assess whether the Group III LysM-RLKs were restricted to the legumes or found more broadly across Angiosperms. RESULTS: Purifying selection was found to be the prevailing form of natural selection within species at LysM-RLKs. No signatures of balancing selection were found in species-wide samples of two wild tomato species. Most genes showed a greater extent of purifying selection in their intracellular domains, with the exception of SlLYK3 which showed strong purifying selection in both the extracellular and intracellular domains in wild tomato species. The phylogenetic analysis did not reveal a clustering of microbe/functional specificity to groups of closely related proteins. We also discovered new putative LysM-III genes in a range of Rosid species, including Eucalyptus grandis. CONCLUSIONS: The LysM-III genes likely originated before the divergence of E. grandis from other Rosids via a fusion of a Group II LysM triplet and a kinase from another RLK family. SlLYK3 emerges as an especially interesting candidate for further study due to the high protein sequence conservation within species, its position in a clade of LysM-RLKs with distinct LysM domains, and its close evolutionary relationship with LYK3 from Arabidopsis thaliana.


Assuntos
Evolução Molecular , Proteínas Serina-Treonina Quinases/genética , Solanum lycopersicum/enzimologia , Solanum lycopersicum/genética , Sequência de Aminoácidos , Filogenia , Domínios Proteicos , Proteínas Serina-Treonina Quinases/química , Proteínas Serina-Treonina Quinases/metabolismo , Seleção Genética , Transcriptoma
5.
Planta ; 249(3): 799-813, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30406411

RESUMO

MAIN CONCLUSION: This study identified biocontrol measures for improving plant quality and resistance under biotic stress caused by the most devastating pathogen in tomato production. The management of plant diseases are dependent on a variety of factors. Two important variables are the soil quality and its bacterial/fungal community. However, the interaction of these factors is not well understood and remains problematic in producing healthy crops. Here, the effect of oak-bark compost, Bacillus subtilis subsp. subtilis, Trichoderma harzianum and two commercial products (FZB24 and FZB42) were investigated on tomato growth, production of metabolites and resistance under biotic stress condition (infection with Phytophthora infestans). Oak-bark compost, B. subtilis subsp. subtilis, and T. harzianum significantly enhanced plant growth and immunity when exposed to P. infestans. However, the commercial products were not as effective in promoting growth, with FZB42 having the weakest protection. Furthermore, elevated levels of anthocyanins did not correlate with enhanced plant resistance. Overall, the most effective and consistent plant protection was obtained when B. subtilis subsp. subtilis was combined with oak-bark compost. In contrast, the combination of T. harzianum and oak-bark compost resulted in increased disease severity. The use of compost in combination with bio-agents should, therefore, be evaluated carefully for a reliable and consistent tomato protection.


Assuntos
Agentes de Controle Biológico/farmacologia , Resistência à Doença , Doenças das Plantas/imunologia , Solanum lycopersicum/crescimento & desenvolvimento , Bacillus subtilis , Compostagem , Solanum lycopersicum/efeitos dos fármacos , Solanum lycopersicum/imunologia , Solanum lycopersicum/microbiologia , Phytophthora infestans , Doenças das Plantas/microbiologia , Microbiologia do Solo , Trichoderma
6.
Environ Microbiol ; 20(1): 30-43, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28967236

RESUMO

Although protists occupy a vast range of habitats and are known to interact with plants among other things via disease suppression, competition or growth stimulation, their contributions to the 'phytobiome' are not well described. To contribute to a more comprehensive picture of the plant holobiont, we examined cercozoan and oomycete taxa living in association with the model plant Arabidopsis thaliana grown in two different soils. Soil, roots, leaves and wooden toothpicks were analysed before and after surface sterilization. Cercozoa were identified using 18S rRNA gene metabarcoding, whereas the Internal Transcribed Spacer 1 was used to determine oomycetes. Subsequent analyses revealed strong spatial structuring of protist communities between compartments, although oomycetes appeared more specialized than Cercozoa. With regards to oomycetes, only members of the Peronosporales and taxa belonging to the genus Globisporangium were identified as shared members of the A. thaliana microbiome. This also applied to cercozoan taxa belonging to the Glissomonadida and Cercomonadida. We identified a strong influence by edaphic factors on the rhizosphere, but not for the phyllosphere. Distinct differences of Cercozoa found preferably in wood or fresh plant material imply specific niche adaptations. Our results highlight the importance of micro-eukaryotes for the plant holobiont.


Assuntos
Arabidopsis/parasitologia , Cercozoários/classificação , Cercozoários/isolamento & purificação , Oomicetos/classificação , Oomicetos/isolamento & purificação , Folhas de Planta/parasitologia , Raízes de Plantas/parasitologia , Cercozoários/genética , DNA Intergênico/genética , Microbiota/fisiologia , Oomicetos/genética , RNA Ribossômico 18S/genética , Rizosfera , Solo/parasitologia
7.
Proc Biol Sci ; 285(1873)2018 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-29491170

RESUMO

Plants possess a battery of specific pathogen resistance (R-)genes. Precise R-gene regulation is important in the presence and absence of a pathogen. Recently, a microRNA family, miR482/2118, was shown to regulate the expression of a major class of R-genes, nucleotide-binding site leucine-rich repeats (NBS-LRRs). Furthermore, RNA silencing suppressor proteins, secreted by pathogens, prevent the accumulation of miR482/2118, leading to an upregulation of R-genes. Despite this transcriptional release of R-genes, RNA silencing suppressors positively contribute to the virulence of some pathogens. To investigate this paradox, we analysed how the regulation of NBS-LRRs by miR482/2118 has been shaped by the coevolution between Phytophthora infestans and cultivated and wild tomatoes. We used degradome analyses and qRT-PCR to evaluate and quantify the co-expression of miR482/2118 and their NBS-LRR targets. Our data show that miR482/2118-mediated targeting contributes to the regulation of NBS-LRRs in Solanum lycopersicum. Based on miR482/2118 expression profiling in two additional tomato species-with different coevolutionary histories with P. infestans-we hypothesize that pathogen-mediated RNA silencing suppression is most effective in the interaction between S. lycopersicum and P. infestans Furthermore, an upregulation of miR482/2118 early in the infection may increase susceptibility to P. infestans.


Assuntos
MicroRNAs/genética , Phytophthora/fisiologia , Doenças das Plantas/genética , RNA de Plantas/genética , Solanum lycopersicum/genética , Solanum lycopersicum/microbiologia , Coevolução Biológica , Resistência à Doença/genética , Doenças das Plantas/microbiologia
8.
Plant Cell Environ ; 41(11): 2530-2548, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-29314046

RESUMO

Plants sense and respond to microbes utilizing a multilayered signalling cascade. In seed plants, the phytohormones jasmonic and salicylic acid (JA and SA) are key denominators of how plants respond to certain microbes. Their interplay is especially well-known for tipping the scales in plants' strategies of dealing with phytopathogens. In non-angiosperm lineages, the interplay is less well understood, but current data indicate that it is intertwined to a lesser extent and the canonical JA/SA antagonism appears to be absent. Here, we used the water fern Azolla filiculoides to gain insights into the fern's JA/SA signalling and the molecular communication with its unique nitrogen fixing cyanobiont Nostoc azollae, which the fern inherits both during sexual and vegetative reproduction. By mining large-scale sequencing data, we demonstrate that Azolla has most of the genetic repertoire to produce and sense JA and SA. Using qRT-PCR on the identified biosynthesis and signalling marker genes, we show that Azolla is responsive to exogenously applied SA. Furthermore, exogenous SA application influenced the abundance and gene expression of Azolla's cyanobiont. Our data provide a framework for JA/SA signalling in ferns and suggest that SA might be involved in Azolla's communication with its vertically inherited cyanobiont.


Assuntos
Ciclopentanos/metabolismo , Gleiquênias/metabolismo , Nostoc/metabolismo , Oxilipinas/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Ácido Salicílico/metabolismo , Gleiquênias/genética , Regulação da Expressão Gênica de Plantas , Fixação de Nitrogênio , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Simbiose
9.
Plant Cell Physiol ; 58(5): 934-945, 2017 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-28340089

RESUMO

The origin of land plants from algae is a long-standing question in evolutionary biology. It is becoming increasingly clear that many characters that were once assumed to be 'embryophyte specific' can in fact be found in their closest algal relatives, the streptophyte algae. One such case is the phenylpropanoid pathway. While biochemical data indicate that streptophyte algae harbor lignin-like components, the phenylpropanoid core pathway, which serves as the backbone of lignin biosynthesis, has been proposed to have arisen at the base of the land plants. Here we revisit this hypothesis using a wealth of new sequence data from streptophyte algae. Tracing the biochemical pathway towards lignin biogenesis, we show that most of the genes required for phenylpropanoid synthesis and the precursors for lignin production were already present in streptophyte algae. Nevertheless, phylogenetic analyses and protein structure predictions of one of the key enzyme classes in lignin production, cinnamyl alcohol dehydrogenase (CAD), suggest that CADs of streptophyte algae are more similar to sinapyl alcohol dehydrogenases (SADs). This suggests that the end-products of the pathway leading to lignin biosynthesis in streptophyte algae may facilitate the production of lignin-like compounds and defense molecules. We hypothesize that streptophyte algae already possessed the genetic toolkit from which the capacity to produce lignin later evolved in vascular plants.


Assuntos
Carofíceas/metabolismo , Lignina/metabolismo , Propanóis/metabolismo , Oxirredutases do Álcool/metabolismo , Evolução Biológica , Interações Hospedeiro-Patógeno
10.
BMC Genomics ; 17(1): 921, 2016 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-27846808

RESUMO

BACKGROUND: Transitions between perennial and an annual life history occur often in plant lineages, but the genes that control whether a plant is an annual or perennial are largely unknown. To identify genes that confer differences between annuals and perennials we compared the gene content of four pairs of sister lineages (Arabidopsis thaliana/Arabidopsis lyrata, Arabis montbretiana/Arabis alpina, Arabis verna/Aubrieta parviflora and Draba nemorosa/Draba hispanica) in the Brassicaceae in which each pair contains one annual and one perennial, plus one extra annual species (Capsella rubella). RESULTS: After sorting all genes in all nine species into gene families, we identified five families in which well-annotated genes are present in the perennials A. lyrata and A. alpina, but are not present in any of the annual species. For the eleven genes in perennials in these families, an orthologous pseudogene or otherwise highly diverged gene was found in the syntenic region of the annual species in six cases. The five candidate families identified encode: a kinase, an oxidoreductase, a lactoylglutathione lyase, a F-box protein and a zinc finger protein. By comparing the active gene in the perennial to the pseudogene or heavily altered gene in the annual, dN and dS were calculated. The low dN/dS values in one kinase suggest that it became pseudogenized more recently, while the other kinase, F-box, oxidoreductase and zinc-finger became pseudogenized closer to the divergence between the annual-perennial pair. CONCLUSIONS: We identified five gene families that may be involved in the life history switch from perennial to annual. Considering the dN and dS data and whether syntenic pseudogenes were found and the potential functions of the genes, the F-box family is considered the most promising candidate for future functional studies to determine if it affects life history.


Assuntos
Arabidopsis/genética , Genes de Plantas , Estudos de Associação Genética , Genoma de Planta , Genômica , Característica Quantitativa Herdável , Arabidopsis/classificação , Biologia Computacional/métodos , Evolução Molecular , Família Multigênica , Filogenia , Pseudogenes
11.
MMWR Morb Mortal Wkly Rep ; 65(29): 739-44, 2016 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-27467820

RESUMO

CDC has updated its interim guidance for U.S. health care providers caring for pregnant women with possible Zika virus exposure, to include the emerging data indicating that Zika virus RNA can be detected for prolonged periods in some pregnant women. To increase the proportion of pregnant women with Zika virus infection who receive a definitive diagnosis, CDC recommends expanding real-time reverse transcription-polymerase chain reaction (rRT-PCR) testing. Possible exposures to Zika virus include travel to or residence in an area with active Zika virus transmission, or sex* with a partner who has traveled to or resides in an area with active Zika virus transmission without using condoms or other barrier methods to prevent infection.(†) Testing recommendations for pregnant women with possible Zika virus exposure who report clinical illness consistent with Zika virus disease(§) (symptomatic pregnant women) are the same, regardless of their level of exposure (i.e., women with ongoing risk for possible exposure, including residence in or frequent travel to an area with active Zika virus transmission, as well as women living in areas without Zika virus transmission who travel to an area with active Zika virus transmission, or have unprotected sex with a partner who traveled to or resides in an area with active Zika virus transmission). Symptomatic pregnant women who are evaluated <2 weeks after symptom onset should receive serum and urine Zika virus rRT-PCR testing. Symptomatic pregnant women who are evaluated 2-12 weeks after symptom onset should first receive a Zika virus immunoglobulin (IgM) antibody test; if the IgM antibody test result is positive or equivocal, serum and urine rRT-PCR testing should be performed. Testing recommendations for pregnant women with possible Zika virus exposure who do not report clinical illness consistent with Zika virus disease (asymptomatic pregnant women) differ based on the circumstances of possible exposure. For asymptomatic pregnant women who live in areas without active Zika virus transmission and who are evaluated <2 weeks after last possible exposure, rRT-PCR testing should be performed. If the rRT-PCR result is negative, a Zika virus IgM antibody test should be performed 2-12 weeks after the exposure. Asymptomatic pregnant women who do not live in an area with active Zika virus transmission, who are first evaluated 2-12 weeks after their last possible exposure should first receive a Zika virus IgM antibody test; if the IgM antibody test result is positive or equivocal, serum and urine rRT-PCR should be performed. Asymptomatic pregnant women with ongoing risk for exposure to Zika virus should receive Zika virus IgM antibody testing as part of routine obstetric care during the first and second trimesters; immediate rRT-PCR testing should be performed when IgM antibody test results are positive or equivocal. This guidance also provides updated recommendations for the clinical management of pregnant women with confirmed or possible Zika virus infection. These recommendations will be updated when additional data become available.


Assuntos
Testes Diagnósticos de Rotina/normas , Surtos de Doenças/prevenção & controle , Guias de Prática Clínica como Assunto , Complicações Infecciosas na Gravidez/prevenção & controle , Infecção por Zika virus/prevenção & controle , Centers for Disease Control and Prevention, U.S. , Feminino , Humanos , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Gravidez , RNA Viral/sangue , Características de Residência/estatística & dados numéricos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Viagem/estatística & dados numéricos , Estados Unidos/epidemiologia , Infecção por Zika virus/transmissão
12.
J Eukaryot Microbiol ; 63(5): 635-41, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27005328

RESUMO

The largest biological surface on earth is formed by plant leaves. These leaf surfaces are colonized by a specialized suite of leaf-inhabiting microorganisms, recently termed "phyllosphere microbiome". Microbial prey, however, attract microbial predators. Protists in particular have been shown to structure bacterial communities on plant surfaces, but virtually nothing is known about the community composition of protists on leaves. Using newly designed specific primers targeting the 18S rDNA gene of Cercozoa, we investigated the species richness of this common protist group on leaves of four Brassicaceae species from two different locations in a cloning-based approach. The generated sequences revealed a broad diversity of leaf-associated Cercozoa, mostly bacterial feeders, but also including known plant pathogens and a taxon of potential endophytes that were recently described as algal predators in freshwater systems. This initial study shows that protists must be regarded as an integral part of the microbial diversity in the phyllosphere of plants.


Assuntos
Biodiversidade , Brassicaceae/parasitologia , Cercozoários/classificação , Cercozoários/genética , Folhas de Planta/parasitologia , Rhizaria/classificação , Rhizaria/genética , Animais , Bactérias , Sequência de Bases , Brassicaceae/classificação , Brassicaceae/microbiologia , Cercozoários/isolamento & purificação , Cercozoários/patogenicidade , Classificação , DNA de Protozoário , DNA Ribossômico/genética , Eucariotos/classificação , Eucariotos/genética , Evolução Molecular , Água Doce/parasitologia , Alemanha , Filogenia , Doenças das Plantas/parasitologia , Folhas de Planta/microbiologia , RNA Ribossômico 18S/genética , Rhizaria/isolamento & purificação
13.
PLoS Genet ; 8(7): e1002813, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22829777

RESUMO

Coevolution between hosts and pathogens is thought to occur between interacting molecules of both species. This results in the maintenance of genetic diversity at pathogen antigens (or so-called effectors) and host resistance genes such as the major histocompatibility complex (MHC) in mammals or resistance (R) genes in plants. In plant-pathogen interactions, the current paradigm posits that a specific defense response is activated upon recognition of pathogen effectors via interaction with their corresponding R proteins. According to the "Guard-Hypothesis," R proteins (the "guards") can sense modification of target molecules in the host (the "guardees") by pathogen effectors and subsequently trigger the defense response. Multiple studies have reported high genetic diversity at R genes maintained by balancing selection. In contrast, little is known about the evolutionary mechanisms shaping the guardee, which may be subject to contrasting evolutionary forces. Here we show that the evolution of the guardee RCR3 is characterized by gene duplication, frequent gene conversion, and balancing selection in the wild tomato species Solanum peruvianum. Investigating the functional characteristics of 54 natural variants through in vitro and in planta assays, we detected differences in recognition of the pathogen effector through interaction with the guardee, as well as substantial variation in the strength of the defense response. This variation is maintained by balancing selection at each copy of the RCR3 gene. Our analyses pinpoint three amino acid polymorphisms with key functional consequences for the coevolution between the guardee (RCR3) and its guard (Cf-2). We conclude that, in addition to coevolution at the "guardee-effector" interface for pathogen recognition, natural selection acts on the "guard-guardee" interface. Guardee evolution may be governed by a counterbalance between improved activation in the presence and prevention of auto-immune responses in the absence of the corresponding pathogen.


Assuntos
Cisteína Proteases/genética , Imunidade Vegetal/genética , Proteínas Secretadas Inibidoras de Proteinases/genética , Seleção Genética/genética , Solanum lycopersicum/genética , Cladosporium/genética , Evolução Molecular , Conversão Gênica , Variação Genética , Interações Hospedeiro-Patógeno/genética , Solanum lycopersicum/parasitologia , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/genética , Polimorfismo Genético
14.
J Clin Microbiol ; 52(1): 67-75, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24153118

RESUMO

A new human coronavirus (CoV), subsequently named Middle East respiratory syndrome (MERS)-CoV, was first reported in Saudi Arabia in September 2012. In response, we developed two real-time reverse transcription-PCR (rRT-PCR) assays targeting the MERS-CoV nucleocapsid (N) gene and evaluated these assays as a panel with a previously published assay targeting the region upstream of the MERS-CoV envelope gene (upE) for the detection and confirmation of MERS-CoV infection. All assays detected ≤10 copies/reaction of quantified RNA transcripts, with a linear dynamic range of 8 log units and 1.3 × 10(-3) 50% tissue culture infective doses (TCID50)/ml of cultured MERS-CoV per reaction. All assays performed comparably with respiratory, serum, and stool specimens spiked with cultured virus. No false-positive amplifications were obtained with other human coronaviruses or common respiratory viral pathogens or with 336 diverse clinical specimens from non-MERS-CoV cases; specimens from two confirmed MERS-CoV cases were positive with all assay signatures. In June 2012, the U.S. Food and Drug Administration authorized emergency use of the rRT-PCR assay panel as an in vitro diagnostic test for MERS-CoV. A kit consisting of the three assay signatures and a positive control was assembled and distributed to public health laboratories in the United States and internationally to support MERS-CoV surveillance and public health responses.


Assuntos
Infecções por Coronavirus/diagnóstico , Coronavirus/classificação , Coronavirus/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Infecções Respiratórias/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Adulto , Coronavirus/genética , Infecções por Coronavirus/virologia , Humanos , Lactente , Recém-Nascido , Kit de Reagentes para Diagnóstico , Infecções Respiratórias/virologia , Estados Unidos , United States Food and Drug Administration
16.
Front Plant Sci ; 14: 1216795, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37965025

RESUMO

Introduction: As key-players of plant immunity, the proteins encoded by resistance genes (R-genes) recognize pathogens and initiate pathogen-specific defense responses. The expression of some R-genes carry fitness costs and therefore inducible immune responses are likely advantageous. To what degree inducible resistance driven by R-genes is triggered by pathogen infection is currently an open question. Methods: In this study we analyzed the expression of 940 R-genes of tomato and potato across 315 transcriptome libraries to investigate how interspecific interactions with microbes influence R-gene expression in plants. Results: We found that most R-genes are expressed at a low level. A small subset of R-genes had moderate to high levels of expression and were expressed across many independent libraries, irrespective of infection status. These R-genes include members of the class of genes called NRCs (NLR required for cell death). Approximately 10% of all R-genes were differentially expressed during infection and this included both up- and down-regulation. One factor associated with the large differences in R-gene expression was host tissue, reflecting a considerable degree of tissue-specific transcriptional regulation of this class of genes. Discussion: These results call into question the widespread view that R-gene expression is induced upon pathogen attack. Instead, a small core set of R-genes is constitutively expressed, imparting upon the plant a ready-to-detect and defend status.

17.
New Phytol ; 196(2): 561-573, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22950721

RESUMO

To sample the natural variation in genes controlling compatibility in the legume-rhizobium symbiosis, we isolated rhizobia from nodules of endemic Lotus species from 21 sites across Europe. The majority of isolates were identified as Mesorhizobium- or Bradyrhizobium-related and formed nitrogen-fixing root nodules on Lotus corniculatus and L. pendunculatus, respectively, thus confirming previously defined cross-inoculation groups. Rhizobium leguminosarum (Rl) strain Norway, isolated from L. corniculatus nodules, displayed an exceptional phenotypic variation on different Lotus genotypes. On L. burttii, Rl Norway formed infected nodules, whereas tumors and elongated infected swellings were induced on L. glaber and L. japonicus ecotype Nepal, respectively. A symbiosis- and Nod-factor-responsive promoter:uidA fusion was strongly and rapidly induced in L. japonicus Gifu, but infection threads or signs of nodule organogenesis were absent. This complex phenotypic pattern was not mimicked by either of three engineered R. leguminosarum bv viciae strains producing different Nod-factor variants. Intriguingly, Rl Norway formed infection threads on Pisum sativum cv Sparkle, but failed to induce organogenesis. Rl Norway thus uncovered variation in symbiotic capabilities among diploid Lotus species and ecotypes that are obscured by optimally adapted M. loti strains. These contrasting infection and organogenesis phenotypes reveal recent diversification of recognition determinants in Lotus.


Assuntos
Interações Hospedeiro-Patógeno/genética , Lotus/genética , Organogênese/genética , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Rhizobium leguminosarum/isolamento & purificação , Nódulos Radiculares de Plantas/microbiologia , Bradyrhizobium/isolamento & purificação , Europa (Continente) , Genótipo , Glucuronidase/genética , Lotus/microbiologia , Mesorhizobium/isolamento & purificação , Fenótipo , Nodulação/genética , Regiões Promotoras Genéticas/genética , Reprodutibilidade dos Testes , Nódulos Radiculares de Plantas/genética , Simbiose/genética , Ativação Transcricional
18.
Protist ; 173(6): 125913, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36257252

RESUMO

In a field experiment we investigated the influence of the environmental filters soil type (i.e. three contrasting soils) and plant species (i.e. lettuce and potato) identity on rhizosphere community assembly of Cercozoa, a dominant group of mostly bacterivorous soil protists. Plant species (14%) and rhizosphere origin (vs bulk soil) with 13%, together explained four times more variation in cercozoan beta diversity than the three soil types (7% explained variation). Our results clearly confirm the existence of plant species-specific protist communities. Network analyses of bacteria-Cercozoa rhizosphere communities identified scale-free small world topologies, indicating mechanisms of self-organization. While the assembly of rhizosphere bacterial communities is bottom-up controlled through the resource supply from root (secondary) metabolites, our results support the hypothesis that the net effect may depend on the strength of top-down control by protist grazers. Since grazing of protists has a strong impact on the composition and functioning of bacteria communities, protists expand the repertoire of plant genes by functional traits, and should be considered as 'protist microbiomes' in analogy to 'bacterial microbiomes'.


Assuntos
Cercozoários , Microbiota , Solo , Microbiologia do Solo , Rizosfera , Bactérias/genética , Eucariotos/genética
19.
Mol Ecol ; 20(13): 2709-23, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21645157

RESUMO

While information about a species' demography is interesting in its own right, it is an absolute necessity for certain types of population genetic analyses. The most widely used methods to infer a species' demographic history do not take intralocus recombination or recent divergence into account, and some methods take several weeks to converge. Here, we present Jaatha, a new composite-likelihood method that does incorporate recent divergence and is also applicable when intralocus recombination rates are high. This new method estimates four demographic parameters. The accuracy of Jaatha is comparable to that of other currently available methods, although it is superior under certain conditions, especially when divergence is very recent. As a proof of concept, we apply this new method to estimate demographic parameters for two closely related wild tomato species, Solanum chilense and S. peruvianum. Our results indicate that these species likely diverged 1.44·N generations ago, where N is the effective population size of S. chilense, and that some introgression between these species continued after the divergence process initiated. Furthermore, S. peruvianum likely experienced a population expansion following speciation.


Assuntos
Biologia Computacional/métodos , Demografia , Modelos Genéticos , Polimorfismo Genético , Solanum/genética , Simulação por Computador/normas , Intervalos de Confiança , DNA de Plantas/química , Fluxo Gênico , Especiação Genética , Funções Verossimilhança , Solanum lycopersicum/classificação , Solanum lycopersicum/genética , Dinâmica Populacional , Recombinação Genética , Tamanho da Amostra , Solanum/classificação , Fatores de Tempo
20.
Eur J Plant Pathol ; 153(1): 1-14, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30880875

RESUMO

All plant species are subject to disease. Plant diseases are caused by parasites, e.g. viruses, bacteria, oomycetes, parasitic plants, fungi, or nematodes. In all organisms, gene expression is tightly regulated and underpins essential functions and physiology. The coordination and regulation of both host and pathogen gene expression is essential for pathogens to infect and cause disease. One mode of gene regulation is RNA silencing. This biological process is widespread in the natural world, present in plants, animals and several pathogens. In RNA silencing, small (20-40 nucleotides) non-coding RNAs (small-RNAs, sRNAs) accumulate and regulate gene expression transcriptionally or post-transcriptionally in a sequence-specific manner. Regulation of sRNA molecules provides a fast mode to alter gene activity of multiple gene transcripts. RNA silencing is an ancient mechanism that protects the most sensitive part of an organism: its genetic code. sRNA molecules emerged as regulators of plant development, growth and plant immunity. sRNA based RNA silencing functions both within and between organisms. Here we present the described sRNAs from plants and pathogens and discuss how they regulate host immunity and pathogen virulence. We speculate on how sRNA molecules can be exploited to develop disease resistant plants. Finally, the activity of sRNA molecules can be prevented by proteins that suppress RNA silencing. This counter silencing response completes the dialog between plants and pathogens controlling plant disease or resistance outcome on the RNA (controlling gene expression) and protein level.

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