RESUMO
Photosynthetic activity is indispensable for plant growth and survival and it depends on the synthesis of plastidial isoprenoids as chlorophylls and carotenoids. In the non-mevalonate pathway (MEP), the 1-deoxy-D-xylulose-5-phosphate synthase 1 (DXS1) enzyme has been postulated to catalyze the rate-limiting step in the formation of plastidial isoprenoids. In tomato, the function of DXS1 has only been studied in fruits, and hence its functional relevance during plant development remains unknown. Here we report the characterization of the wls-2297 tomato mutant, whose severe deficiency in chlorophylls and carotenoids promotes an albino phenotype. Additionally, growth of mutant seedlings was arrested without developing vegetative organs, which resulted in premature lethality. Gene cloning and silencing experiments revealed that the phenotype of wls-2297 mutant was caused by 38.6 kb-deletion promoted by a single T-DNA insertion affecting the DXS1 gene. This was corroborated by in vivo and molecular complementation assays, which allowed the rescue of mutant phenotype. Further characterization of tomato plants overexpressing DXS1 and comparative expression analysis indicate that DXS1 may play other important roles besides to that proposed during fruit carotenoid biosynthesis. Taken together, these results demonstrate that DXS1 is essentially required for the development and survival of tomato plants.
Assuntos
DNA Bacteriano/metabolismo , Proteínas de Plantas/metabolismo , Solanum lycopersicum/enzimologia , Transferases/metabolismo , Carotenoides/metabolismo , Clorofila/metabolismo , Clonagem Molecular , DNA Bacteriano/genética , DNA Complementar/metabolismo , Frutas/química , Frutas/metabolismo , Solanum lycopersicum/crescimento & desenvolvimento , Mutagênese , Fenótipo , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Proteínas de Plantas/antagonistas & inibidores , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Interferência de RNA , Plântula/crescimento & desenvolvimento , Transferases/antagonistas & inibidores , Transferases/genéticaRESUMO
Pentacyclic triterpenes are gaining interest due to their beneficial health effects, as anti-inflammatory, anti-diabetic and anti-tumoral, among others. In this study, an analytical LC-MS method was developed for the simultaneous determination of maslinic, oleanolic and ursolic acids along with erythrodiol and uvaol, which are the main triterpenic compounds present in the fruits and leaves of Olea europaea L. A Zorbax Eclipse PAH column at 30°C with mobile phase of water (17%) and methanol (83%) at 0.8mL/min conformed the optimal chromatographic conditions that allowed the separation of the compounds of interest, two pairs of which are isomers differing only in the position of one methyl group (oleanolic-ursolic and erythrodiol-uvaol). The ionization was performed in an APCI source at 450°C programmed in negative mode for the triterpenic acids, and in positive for the alcohols. An ion trap (LC-IT-MS) and a triple quadrupole (LC-QqQ-MS) were assessed for maximal sensitivity that was achieved with LC-QqQ-MS. The LODs of triterpenic acids were lower than 1nM, whereas for erythrodiol and uvaol were 4.5 and 7.5nM, respectively. The method was linear for the five analytes in the range of concentrations from 0.005 to 15µM with correlation coefficients exceeding 0.99. The precision and accuracy were ≤9.90% and ≤9.57%, respectively. The applicability of the validated method was assessed in the analysis of the pentacyclic triterpenes in Marfil table olives, after the optimization of the extraction procedure. The developed method constitutes the first step for future studies of triterpenic compounds present in foods that would allow establishing their effects on human health.
Assuntos
Olea/química , Triterpenos Pentacíclicos/análise , Cromatografia Líquida de Alta Pressão/métodos , Frutas/química , Limite de Detecção , Espectrometria de Massas/métodos , Extratos Vegetais/química , Folhas de Planta/química , EspanhaRESUMO
Reacciones adversas a los medicamentos: acercamiento epidemiológico- Reacciones adversas a excipientes- Sistema Nacional de Farmacovigilancia en Argentina- Botiquín de urgencias en intoxicaciones- Farmacodependencia- Prevención comunitaria de adicciones- Detección de problemas galénicos en medicamentos- Interacciones medicamentosas- Interferencias de medicamentos en el resultado de pruebas de laboratorio- Hepatopatías e insuficiencia renal: prescripción- Monitoreo de sustancias farmacológicas activas en sangre- Listas oficiales: Estupefacientes y psicotrópicos (I-IV); Sustancias no permitidas en actividades deportivas- Ordenamiento completo de principios activos, contraindicaciones y reacciones adversas medicamentosas
Assuntos
Dicionários Farmacêuticos como Assunto , Doença Medicamentosa , Erros de Medicação/classificação , Interações Medicamentosas , Medicamentos Essenciais , Prescrições de Medicamentos/normas , Transtornos Relacionados ao Uso de Substâncias/prevenção & controle , Tratamento Farmacológico , Vigilância de Produtos Comercializados , Analgésicos Opioides , Catálogos de Medicamentos como Assunto , Controle de Medicamentos e Entorpecentes/legislação & jurisprudência , Controle de Medicamentos e Entorpecentes/métodos , Controle de Qualidade , Dopagem Esportivo , Entorpecentes , Excipientes Farmacêuticos/efeitos adversos , Hepatopatias , Insuficiência Renal Crônica , Legislação de Medicamentos , Injúria Renal Aguda , Monitoramento de Medicamentos/normas , Psicotrópicos , Sistemas de Notificação de Reações Adversas a Medicamentos , Transtornos Relacionados ao Uso de Substâncias , Transtornos Relacionados ao Uso de Substâncias/classificação , Transtornos Relacionados com Narcóticos , Tratamento Farmacológico/efeitos adversos , Técnicas de Laboratório Clínico/efeitos adversos , Técnicas de Laboratório Clínico/normas , Veículos HomeopáticosRESUMO
Reacciones adversas a los medicamentos: acercamiento epidemiológico- Reacciones adversas a excipientes- Sistema Nacional de Farmacovigilancia en Argentina- Botiquín de urgencias en intoxicaciones- Farmacodependencia- Prevención comunitaria de adicciones- Detección de problemas galénicos en medicamentos- Interacciones medicamentosas- Interferencias de medicamentos en el resultado de pruebas de laboratorio- Hepatopatías e insuficiencia renal: prescripción- Monitoreo de sustancias farmacológicas activas en sangre- Listas oficiales: Estupefacientes y psicotrópicos (I-IV); Sustancias no permitidas en actividades deportivas- Ordenamiento completo de principios activos, contraindicaciones y reacciones adversas medicamentosas