Methods for the assessment of isoenzymes activity in sunflower seeds

Abstract

-Enzyme activity detection in seeds allows the assessment of the deterioration process in the early stages. In this context, the objective was to assess the effectiveness and the degree of similarity between the results of izoenzymes activity using different methods, as follows: spectrophotometry, visual analysis after electrophoresis, evaluation of the bands nodes gels with software GelAnalyzer. 250 hybrid seeds were submitted to artificial aging to form four lots with differences in physiological quality. In addition to the enzymatic determinations, physiological germination tests, emergence and emergence speed index were performed. It was concluded that there are differences in results according to the methods used; the results were consistent between test methods and correlated with physiological results when the catalase enzyme was evaluated; and in the isozyme analysis of superoxide dismutase and alcohol dehydrogenase there was a divergence among the results of the methodologies, and according to such, the relationship to seeds physiological quality.Index terms: seed quality, deterioration, enzymatic changes.Métodos para a avaliação da atividade de isoenzimas em sementes de girassol RESUMO -A detecção da atividade enzimática em sementes permite avaliar o processo de deterioração ainda nos primeiros estádios. Nesse contexto, objetivou-se avaliar a similaridade entre os resultados da atividade de isoenzimas, utilizando-se diferentes métodos: espectrofotometria, análise visual dos géis de poliacrilamida após eletroforese e avaliação das bandas nós géis com auxílio do software GelAnalyzer, e sua relação com a qualidade fisiológica das sementes. Foram utilizadas sementes do híbrido 250 da empresa Heliagro submetidas ao envelhecimento artificial para a constituição de quatro sublotes com diferenças na qualidade fisiológica. Além das determinações enzimáticas, foram realizados os testes fisiológicos de germinação, emergência e índice de velocidade de emergência. Concluiu-se que há diferenças de resultados conforme o método utilizado; os resultados foram consistentes entre métodos de análise e correlacionados com os resultados fisiológicos quando a enzima avaliada foi a catalase; para a análise isoenzimática da superóxido dismutase e da álcool desidrogenase houve divergência entre os resultados das metodologias, e em função dessas a relação com a qualidade fisiológica das sementes.Termos para indexação: qualidade de sementes, deterioração, alterações enzimáticas.

Oxidative and Fibrinolytic Mechanisms: Two Important Processes to Consider in Platelet Storage

“…In our study, we were unable to demonstrate differences in SOD activity in platelet lysates, and this failure may have been due to the fact that the concentration of SOD in this blood component is approximately 1 fg of SOD per platelet with 77% in the Cu/Zn isoform, while the rest is in the Mn-SOD isoform, and this is insufficient for evaluation by zymography. However, Faria et al [ 31 ] evaluated the enzymatic activity of SOD using two different techniques and concluded that there were differences in the evaluation results of SOD activity as well as alcohol dehydrogenase between the two methodological techniques used. Better sensitivity of enzymatic evaluations by spectrophotometric techniques was reported [ 31 , 32 , 33 ].…”

“…However, Faria et al [ 31 ] evaluated the enzymatic activity of SOD using two different techniques and concluded that there were differences in the evaluation results of SOD activity as well as alcohol dehydrogenase between the two methodological techniques used. Better sensitivity of enzymatic evaluations by spectrophotometric techniques was reported [ 31 , 32 , 33 ].…”

Herbicides Tolerance in a Pseudomonas Strain Is Associated With Metabolic Plasticity of Antioxidative Enzymes Regardless of Selection

“…The gels were exposed to white light and immersed in water until the bands corresponding to the SOD bands became visible. Since there are differences between measurements of enzymatic activities ( Faria et al, 2015 ), in the case of SOD we prefer to use the electrophoretic method to the enzymatic one.…”

Sunflower seed treatment with growth inhibitor: Crop development aspects and yield

“…Considering the higher resistance to heat, drought and cold than most of the species grown in Brazil (Lima et al, 2014), sunflower areas expanded with about 25% in 2014/2015 as compared to the agricultural year of 2013/2014 (CONAB, 2015). This crop has received more attention due to its good results as an option to succession planting and crop rotation, animal feed, honey production and functional human nutrition (Faria et al, 2015;Ungaro et al, 2009). Furthermore, since 2010, the Brazilian government demands that all diesel commercialized in Brazil should be admixed with at least 5% of biodiesel (Matsumoto et al, 2015).…”