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1.
J Proteome Res ; 17(9): 3114-3127, 2018 09 07.
Artículo en Inglés | MEDLINE | ID: mdl-30084641

RESUMEN

Plum pox virus (PPV, family Potyviridae) is one of the most important viral pathogens of Prunus spp. causing considerable damage to stone-fruit industry worldwide. Among the PPV strains identified so far, only PPV-C, PPV-CR, and PPV-CV are able to infect cherries under natural conditions. Herein, we evaluated the pathogenic potential of two viral isolates in herbaceous host Nicotiana benthamiana. Significantly higher accumulation of PPV capsid protein in tobacco leaves infected with PPV-CR (RU-30sc isolate) was detected in contrast to PPV-C (BY-101 isolate). This result correlated well with the symptoms observed in the infected plants. To further explore the host response upon viral infection at the molecular level, a comprehensive proteomic profiling was performed. Using reverse-phase ultra-high-performance liquid chromatography followed by label-free mass spectrometry quantification, we identified 38 unique plant proteins as significantly altered due to the infection. Notably, the abundances of photosynthesis-related proteins, mainly from the Calvin-Benson cycle, were found more aggressively affected in plants infected with PPV-CR isolate than those of PPV-C. This observation was accompanied by a significant reduction in the amount of photosynthetic pigments extracted from the leaves of PPV-CR infected plants. Shifts in the abundance of proteins that are involved in stimulation of photosynthetic capacity, modification of amino acid, and carbohydrate metabolism may affect plant growth and initiate energy formation via gluconeogenesis in PPV infected N. benthamiana. Furthermore, we suggest that the higher accumulation of H2O2 in PPV-CR infected leaves plays a crucial role in plant defense and development by activating the glutathione synthesis.


Asunto(s)
Regulación de la Expresión Génica de las Plantas , Proteínas de Choque Térmico/genética , Nicotiana/genética , Enfermedades de las Plantas/genética , Proteínas de Plantas/genética , Virus Eruptivo de la Ciruela/patogenicidad , Carotenoides/biosíntesis , Clorofila/biosíntesis , Cromatografía Líquida de Alta Presión , Cromatografía de Fase Inversa , Metabolismo Energético/genética , Genotipo , Glutatión/biosíntesis , Proteínas de Choque Térmico/clasificación , Proteínas de Choque Térmico/metabolismo , Interacciones Huésped-Patógeno/genética , Peróxido de Hidrógeno/metabolismo , Espectrometría de Masas , Oxidación-Reducción , Fotosíntesis/genética , Enfermedades de las Plantas/virología , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Hojas de la Planta/virología , Proteínas de Plantas/clasificación , Proteínas de Plantas/metabolismo , Virus Eruptivo de la Ciruela/clasificación , Virus Eruptivo de la Ciruela/genética , Virus Eruptivo de la Ciruela/crecimiento & desarrollo , Prunus avium/virología , Prunus domestica/virología , Nicotiana/metabolismo , Nicotiana/virología
2.
J Agric Food Chem ; 62(47): 11547-56, 2014 Nov 26.
Artículo en Inglés | MEDLINE | ID: mdl-25365400

RESUMEN

Although ginkgo (Maidenhair tree, Ginkgo biloba L.) is an ancient medicinal and ornamental tree, there has not previously been any systematic proteomic study of the leaves. Herein we describe results from the initial study identifying abundant ginkgo leaf proteins and present a gel reference map. Proteins were isolated from fully developed mature leaves in biological triplicate and analyzed by two-dimensional electrophoresis plus tandem mass spectrometry. Using this approach, we were able to reproducibly quantify 190 abundant protein spots, from which 157 proteins were identified. Most of identified proteins are associated with the energy and protein destination/storage categories. The reference map provides a basis for understanding the accumulation of flavonoids and other phenolic compounds in mature leaves (e.g., identification of chalcone synthase, the first committed enzyme in flavonoid biosynthesis). We additionally detected several proteins of as yet unknown function. These proteins comprise a pool of potential targets that might be useful in nontraditional medical applications.


Asunto(s)
Ginkgo biloba/química , Hojas de la Planta/química , Proteoma/metabolismo , Cromatografía Liquida , Electroforesis en Gel Bidimensional , Flavonoides/análisis , Fenoles/análisis , Proteínas de Plantas/química , Proteómica , Espectrometría de Masas en Tándem
3.
J Proteome Res ; 12(11): 4799-806, 2013 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-24111740

RESUMEN

Starting in 2007, we have grown soybean (Glycine max [L.] Merr. variety Soniachna) and flax (Linum usitatissimum, L. variety Kyivskyi) in the radio-contaminated Chernobyl area and analyzed the seed proteomes. In the second-generation flax seeds, we detected a 12% increase in oil content. To characterize the bases for this increase, seed development has been studied. Flax seeds were harvested in biological triplicate at 2, 4, and 6 weeks after flowering and at maturity from plants grown in nonradioactive and radio-contaminated plots in the Chernobyl area for two generations. Quantitative proteomic analyses based on 2-D gel electrophoresis (2-DE) allowed us to establish developmental profiles for 199 2-DE spots in both plots, out of which 79 were reliably identified by tandem mass spectrometry. The data suggest a statistically significant increased abundance of proteins associated with pyruvate biosynthesis via cytoplasmic glycolysis, L-malate decarboxylation, isocitrate dehydrogenation, and ethanol oxidation to acetaldehyde in early stages of seed development. This was followed by statistically significant increased abundance of ketoacyl-[acylcarrier protein] synthase I related to condensation of malonyl-ACP with elongating fatty acid chains. On the basis of these and previous data, we propose a preliminary model for plant adaptation to growth in a radio-contaminated environment. One aspect of the model suggests that changes in carbon assimilation and fatty acid biosynthesis are an integral part of plant adaptation.


Asunto(s)
Adaptación Biológica/efectos de la radiación , Accidente Nuclear de Chernóbil , Lino/genética , Lino/efectos de la radiación , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Modelos Biológicos , Proteoma/efectos de la radiación , Adaptación Biológica/genética , Carbono/metabolismo , Radioisótopos de Cesio/análisis , Electroforesis en Gel Bidimensional , Lino/química , Regulación de la Expresión Génica de las Plantas/genética , Glucólisis , Aceite de Linaza/análisis , Proteoma/genética , Ácido Pirúvico/metabolismo , Semillas/química , Semillas/crecimiento & desarrollo , Semillas/efectos de la radiación , Suelo/química , Radioisótopos de Estroncio/análisis , Espectrometría de Masas en Tándem , Ucrania
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