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1.
Methods Mol Biol ; 1631: 313-323, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28735407

RESUMEN

Polyamines are polycationic nitrogenous compounds that accumulate in plants exposed to abiotic stresses. In higher animals, they influence gene expression, brain development, and nerve growth and regeneration. Because of their known roles in plant stress responses, quantitative determination of polyamines is very important. Polyamines in their native form cannot be detected by optical and/or electrochemical methods as they do not show any structural features, and hence derivatization of polyamines is essential to make them to produce either chromophores or fluorescence. Here we describe various methods of derivatization using different labeling agents and suitable separation and detecting methods for a vast source of polyamines existing in living cells.


Asunto(s)
Electroforesis Capilar/métodos , Poliaminas/análisis , Animales , Plantas , Poliaminas/química
2.
Plant Cell Rep ; 34(1): 141-56, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25348337

RESUMEN

KEY MESSAGE: Polyamines can regulate the expression of antioxidant enzymes and impart plants tolerance to abiotic stresses. A comparative analysis of polyamines, their biosynthetic enzymes at kinetic and at transcriptional level, and their role in regulating the induction of antioxidant defense enzymes under salt stress condition in two foxtail millet (Setaria italica L.) cultivars, namely Prasad, a salt-tolerant, and Lepakshi, a salt-sensitive cultivar was conducted. Salt stress resulted in elevation of free polyamines due to increase in the activity of spermidine synthase and S-adenosyl methionine decarboxylase enzymes in cultivar Prasad compared to cultivar Lepakshi under different levels of NaCl stress. These enzyme activities were further confirmed at the transcript level via qRT-PCR analysis. The cultivar Prasad showed a greater decrease in diamine oxidase and polyamine oxidase activity, which results in the accumulation of polyamine pools over cultivar Lepakshi. Generation of free radicals, such as O 2 (·-) and H2O2, was also analyzed quantitatively. A significant increase in O 2 (·-) and H2O2 in the cultivar Lepakshi compared with cultivar Prasad was recorded in overall pool sizes. Further, histochemical staining showed lesser accumulation of O 2 (·-) and of H2O2 in the leaves of cultivar Prasad than cultivar Lepakshi. Our results also suggest the ability of polyamine oxidation in regulating the induction of antioxidative defense enzymes, which involve in the elimination of toxic levels of O 2 (·-) and H2O2, such as Mn-superoxide dismutase, catalase and ascorbate peroxidase. The contribution of polyamines in modulating antioxidative defense mechanism in NaCl stress tolerance is discussed.


Asunto(s)
Antioxidantes/metabolismo , Poliaminas/metabolismo , Tolerancia a la Sal , Setaria (Planta)/metabolismo , Adenosilmetionina Descarboxilasa/genética , Adenosilmetionina Descarboxilasa/metabolismo , Catalasa/genética , Catalasa/metabolismo , Relación Dosis-Respuesta a Droga , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Peróxido de Hidrógeno/metabolismo , Oxidorreductasas actuantes sobre Donantes de Grupo CH-NH/genética , Oxidorreductasas actuantes sobre Donantes de Grupo CH-NH/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Plantones/genética , Plantones/metabolismo , Setaria (Planta)/clasificación , Setaria (Planta)/genética , Cloruro de Sodio/farmacología , Especificidad de la Especie , Espermidina Sintasa/genética , Espermidina Sintasa/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Superóxidos/metabolismo , Poliamino Oxidasa
3.
J Plant Physiol ; 168(7): 671-7, 2011 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-21035898

RESUMEN

Plants produce an array of proteins as a part of a global response to protect the cell metabolism when they grow under environmental conditions such as drought and salinity that generate reduced water potential. The synthesis of hydrophilic proteins is a major part of the response to water deficit conditions. An increased expression of LEA proteins is thought to be one of the primary lines of defense to prevent the loss of intercellular water during adverse conditions. These LEA proteins are known to prevent aggregation of a wide range of other proteins. In this study we report the water stress induced protein aggregation and its abrogation followed by expression of group 1 and group 2 LEA proteins of water soluble proteomes in horsegram. Water stress caused an increased protein aggregation with magnitude and duration of stress in horsegram seedlings. Tissue-specific expression of LEA 1 protein decreased in the embryonic axis when compared to cotyledons in 24h stressed seedlings. We found no cross reaction of LEA 1 with proteome of 48h stressed embryonic axis and 72h stressed root and shoot samples. However, LEA 2 antibodies were cross reacted with four polypeptides with different molecular mass in shoot tissue samples and found no reaction with root proteome as evidenced from immuno-blot analysis. The role of LEA proteins in relation to protein aggregation during water stressed conditions was discussed.


Asunto(s)
Deshidratación/metabolismo , Fabaceae/metabolismo , Germinación/fisiología , Proteínas de Plantas/metabolismo , Plantones/metabolismo , Western Blotting , Electroforesis , Fabaceae/fisiología , Plantones/fisiología
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