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1.
Phytopathology ; 107(5): 519-527, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-28112596

RESUMO

Prevalence of Xanthomonas translucens, which causes cereal leaf streak (CLS) in cereal crops and bacterial wilt in forage and turfgrass species, has increased in many regions in recent years. Because the pathogen is seedborne in economically important cereals, it is a concern for international and interstate germplasm exchange and, thus, reliable and robust protocols for its detection in seed are needed. However, historical confusion surrounding the taxonomy within the species has complicated the development of accurate and reliable diagnostic tools for X. translucens. Therefore, we sequenced genomes of 15 X. translucens strains representing six different pathovars and compared them with additional publicly available X. translucens genome sequences to obtain a genome-based phylogeny for robust classification of this species. Our results reveal three main clusters: one consisting of pv. cerealis, one consisting of pvs. undulosa and translucens, and a third consisting of pvs. arrhenatheri, graminis, phlei, and poae. Based on genomic differences, diagnostic loop-mediated isothermal amplification (LAMP) primers were developed that clearly distinguish strains that cause disease on cereals, such as pvs. undulosa, translucens, hordei, and secalis, from strains that cause disease on noncereal hosts, such as pvs. arrhenatheri, cerealis, graminis, phlei, and poae. Additional LAMP assays were developed that selectively amplify strains belonging to pvs. cerealis and poae, distinguishing them from other pathovars. These primers will be instrumental in diagnostics when implementing quarantine regulations to limit further geographic spread of X. translucens pathovars.


Assuntos
Genoma Bacteriano/genética , Genômica , Doenças das Plantas/microbiologia , Xanthomonas/classificação , Primers do DNA/genética , Técnicas de Amplificação de Ácido Nucleico , Filogenia , Sensibilidade e Especificidade , Especificidade da Espécie , Xanthomonas/genética , Xanthomonas/isolamento & purificação
2.
Appl Environ Microbiol ; 80(15): 4519-30, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24837384

RESUMO

Molecular diagnostics for crop diseases can enhance food security by enabling the rapid identification of threatening pathogens and providing critical information for the deployment of disease management strategies. Loop-mediated isothermal amplification (LAMP) is a PCR-based tool that allows the rapid, highly specific amplification of target DNA sequences at a single temperature and is thus ideal for field-level diagnosis of plant diseases. We developed primers highly specific for two globally important rice pathogens, Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight (BB) disease, and X. oryzae pv. oryzicola, the causal agent of bacterial leaf streak disease (BLS), for use in reliable, sensitive LAMP assays. In addition to pathovar distinction, two assays that differentiate X. oryzae pv. oryzae by African or Asian lineage were developed. Using these LAMP primer sets, the presence of each pathogen was detected from DNA and bacterial cells, as well as leaf and seed samples. Thresholds of detection for all assays were consistently 10(4) to 10(5) CFU ml(-1), while genomic DNA thresholds were between 1 pg and 10 fg. Use of the unique sequences combined with the LAMP assay provides a sensitive, accurate, rapid, simple, and inexpensive protocol to detect both BB and BLS pathogens.


Assuntos
Técnicas de Amplificação de Ácido Nucleico/métodos , Oryza/microbiologia , Doenças das Plantas/microbiologia , Xanthomonas/isolamento & purificação , Primers do DNA/genética , DNA Bacteriano/genética , Sensibilidade e Especificidade , Xanthomonas/classificação , Xanthomonas/genética
3.
Bioorg Med Chem Lett ; 24(15): 3552-5, 2014 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-24915877

RESUMO

Here, we describe the synthesis of two new fluorescent derivatives of thiamethoxam and compared their toxicity on aphid Acyrthosiphon pisum and their mode of action on insect nicotinic acetylcholine receptors expressed on the sixth abdominal ganglion. The compound 3 with two 2-chlorothiazole moieties was found to be more toxic using toxicological bioassays 24 h and 48 h after exposure while compound 4 appeared more active using cockroach ganglionic depolarization. Interestingly, thiamethoxam appeared more effective than component 3 and 4, respectively. Our results demonstrated that component 3 and 4 act as agonists of insect nicotinic acetylcholine receptors.


Assuntos
Afídeos/efeitos dos fármacos , Corantes Fluorescentes/farmacologia , Inseticidas/farmacologia , Nitrocompostos/farmacologia , Oxazinas/farmacologia , Receptores Nicotínicos/metabolismo , Tiazóis/farmacologia , Animais , Relação Dose-Resposta a Droga , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/química , Inseticidas/síntese química , Inseticidas/química , Conformação Molecular , Neonicotinoides , Nitrocompostos/síntese química , Nitrocompostos/química , Oxazinas/síntese química , Oxazinas/química , Relação Estrutura-Atividade , Tiametoxam , Tiazóis/síntese química , Tiazóis/química
4.
Nat Biomed Eng ; 4(2): 172-180, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31792423

RESUMO

Retinal dystrophies and age-related macular degeneration related to photoreceptor degeneration can cause blindness. In blind patients, although the electrical activation of the residual retinal circuit can provide useful artificial visual perception, the resolutions of current retinal prostheses have been limited either by large electrodes or small numbers of pixels. Here we report the evaluation, in three awake non-human primates, of a previously reported near-infrared-light-sensitive photovoltaic subretinal prosthesis. We show that multipixel stimulation of the prosthesis within radiation safety limits enabled eye tracking in the animals, that they responded to stimulations directed at the implant with repeated saccades and that the implant-induced responses were present two years after device implantation. Our findings pave the way for the clinical evaluation of the prosthesis in patients affected by dry atrophic age-related macular degeneration.


Assuntos
Degeneração Macular/reabilitação , Movimentos Sacádicos , Visão Ocular/fisiologia , Percepção Visual , Próteses Visuais , Animais , Modelos Animais de Doenças , Medições dos Movimentos Oculares , Macaca fascicularis , Degeneração Macular/fisiopatologia , Masculino , Estimulação Luminosa , Células Ganglionares da Retina/fisiologia
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