ABSTRACT
Stripe rust (Puccinia striiformis f.sp. tritici) is the most devastating disease of wheat (Triticum aestivum L.) accounting huge economical losses to the industry worldwide. HD 2329 was a widely grown wheat cultivar which had become highly susceptible to stripe rust and was used to understand the biochemical aspects of the host pathogen interaction through characterization of superoxide dismutase (SOD). In the present study, two types of SOD, ionically or covalently bound to the particulate fraction were found in the stripe rust infected and uninfected wheat leaves of susceptible cultivar HD 2329. Cell walls of leaves contained a high level of SOD, of which 41-44% was extractable by 2 M NaCl and 10-13% by 0.5% EDTA in infected and uninfected leaves. The NaCl-released SOD constituted the predominant fraction. It exhibited maximum activity at pH 9.0, had a Km value of 1.82-2.51 for uninfected and 1.77-2.37 mM for infected, respectively with pyrogallol as the substrate, and a Vmax of 9.55-21.4 and 12.4-24.1 A min-1g-1FW. A temperature optimum of 20oC was observed for SOD of both uninfected and infected leaves. SOD showed differential response to metal ions, suggesting their distinctive nature. Inhibition of wall bound SOD by iodine and its partial regeneration of activity by mercaptoethanol suggested the involvement of cysteine in active site of the enzyme. These two forms showed greater differences with respect to thermodynamic properties like energy of activation (Ea) and enthalpy change (H), while entropy change (S) and free energy change (G) were similar. The results further showed that pathogen infection of the leaves of susceptible wheat cultivar induced a decrease in the SOD activity and kinetics which might be critical during the response of plant cells to the infection.
Subject(s)
Basidiomycota/metabolism , Basidiomycota/pathogenicity , Cell Wall/chemistry , Cell Wall/enzymology , Cell Wall/metabolism , Enzyme Inhibitors/chemistry , Hydrogen-Ion Concentration , Kinetics , Metals/chemistry , Plant Cells/enzymology , Plant Diseases/microbiology , Plant Leaves/enzymology , Superoxide Dismutase/chemistry , Superoxide Dismutase/pharmacokinetics , Temperature , Triticum/enzymologyABSTRACT
Se efectuó la standarización de la rata Sprague-Dawley en relación a parámetros hematológicos, bioquímicos y anatómicos en una muestra de 24 ratas, 12 machos y 12 hembras. Se estandarizó el modelo de asa duodenal cerrada para producir pancreatitis aguda en 72 ratas, las que fueron sacrificadas en grupos de 3 cada hora durante 24 horas. Se determinó que el momento en que se inicia la pancreatitis aguda, tanto por parámetros bioquímicos como histopatológicos, corresponde a la sexta hora postinducción. Utilizando este modelo se efectuaron diferentes acciones tendientes a disminuir los efectos de una PA de inicio reciente. Cien ratas fueron divididas en 5 grupos de 20: 1. Control, 2. Descompresión biliar precoz, 3. Verapamilo 0,5 mg/kg ev., 4. Allopurinol 50 mg/kg ev., 5. Superóxido dismutasa 5 mg/kg ev. Para determinar el efecto benéfico de estas acciones sobre la PA, ellas fueron realizadas a la sexta hora postinducción. Se concluye que los mejores efectos logrados en este modelo corresponden a la desobtrucción efectuada precozmente. Dentro de las drogas utilizadas, como drogas captadoras de radicales libres de oxígeno, la superóxido dismutasa fue la que presentó los mejores efectos benéficiosos. El allopurinol mostró efectos discretos en este modelo, no comprobándose el efecto beneficioso del verapamilo en relación al grupo control