Functional Analysis of Durum Wheat GASA1 Protein as a Biotechnological Alternative Against Plant Fungal Pathogens and a Positive Regulator of Biotic Stress Defense

Plants are frequently challenged by a variety of microorganisms. To protect themselves against harmful invaders, they have evolved highly effective defense mechanisms, including the synthesis of numerous types of antimicrobial peptides (AMPs). Snakins are such compounds, encoded by the GASA (Gibberellic Acid-Stimulated Arabidopsis) gene family, and are involved in the response to biotic and abiotic stress. Here, we examined the function of the newly identified TdGASA1 gene and its encoded protein in Triticum durum subjected to different biotic stress-related simulants, such as mechanical injury, methyl jasmonate (MeJA), indole-3-acetic acid (IAA), salicylic acid (SA), hydrogen peroxide (H₂O₂), as well as infection with pathogenic fungi Fusarium graminearum and Aspergillus niger. We found that in durum wheat, TdGASA1 transcripts were markedly increased in response to these stress simulants. Isolated and purified TdGASA1 protein exhibited significant antifungal activity in the growth inhibition test conducted on eight species of pathogenic fungi on solid and liquid media. Transgenic Arabidopsis lines overexpressing TdGASA1 obtained in this study showed higher tolerance to detrimental effects of H₂O₂, MeJA, and ABA treatment. In addition, these lines exhibited resistance to Fusarium graminearum and Aspergillus niger, which was linked to a marked increase in antioxidant activity in the leaves under stress conditions. This resistance was correlated with the upregulation of pathogenesis-related genes (AtPDF1.2a, AtERF1, AtVSP2, AtMYC2, AtPR1, AtACS6, AtETR1, and AtLOX2) in the transgenic lines. Overall, our results indicate that TdGASA1 gene and its encoded protein respond ubiquitously to a range of biotic stimuli and seem to be crucial for the basal resistance of plants against pathogenic fungi. This gene could therefore be a valuable target for genetic engineering to enhance wheat resistance to biotic stress.