Evaluation of Superoxide Dismutase Isoforms Activity and Defense System-Related Proteins’ Expression in Ascochyta Blight-Infected Chickpea Using 2D Electrophoresis Technique

Abstract

Fungal diseases are crucial factors in reducing chickpea production. Ascochyta Blight is caused by the necrotrophic fungi Didymella rabiei and is one of the most destructive diseases in most world areas. Therefore, a completely randomized factorial design with five replications was applied to evaluate the effects of Ascochyta Blight disease fungi on the chickpea plant. The chlorophyll index, the activities of superoxide dismutase isoforms, and evolutionary analyses were performed to get further insights. Also, 2D electrophoresis of chickpea leaf proteins, gene ontology, and protein-protein interactions analysis was performed. The results did not show any significant effect of A. rabiei infection on the wet weight of chickpea seedlings. Chlorophyll index levels significantly decreased with A. rabiei infection in both chickpea lines. Electrophoretic analysis of superoxide dismutase on 8% polyacrylamide gel revealed three isoforms. The activities of superoxide dismutase isoforms significantly increased with A. rabiei disease. Identification of proteins was performed according to their isoelectric points and approximate molecular weights. Leaf proteome analysis of chickpea lines showed that the expression of eight reproducible spots changed significantly under A. rabiei disease condition. Candidate proteins were components of defense and regulation systems. High expression of Dual specificity protein, Peptide methionine sulfoxide reductase B2, and chloroplastic phosphatase 1B (the proteins involved in the defense system) reveals their essential functions under A. rabiei infection. Increased activities of superoxide dismutase enzymes and proteins involved in the defense system can reduce A. rabiei infection effects on chickpea seedlings.