Kinetic properties of cell wall bound superoxide dismutase in leaves of wheat (<i>Triticum aestivum</i> L.) following stripe rust (<i>Puccinia striiformis</i>) infection

Abstract

Stripe rust (<i>Puccinia striiformis</i> f.sp. <i>tritici</i>) is the most devastating disease of wheat (<i>Triticum aestivum</i> 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<i> </i>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 <i>K</i>_m value of 1.82-2.51 for uninfected and 1.77-2.37 mM for infected, respectively with pyrogallol as the substrate, and a <i>V</i>_max of 9.55-21.4 and 12.4-24.1 <img src='/image/spc_char/delta.gif' border=0>A min^-1g^-1FW. A temperature optimum of 20^oC 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 (E_a) and enthalpy change (<img src='/image/spc_char/delta.gif' border=0>H), while entropy change (<img src='/image/spc_char/delta.gif' border=0>S) and free energy change (<img src='/image/spc_char/delta.gif' border=0>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.