DEVELOPMENT OF STEM RUST RESISTANT TRANSGENIC WHEAT PLANTS THROUGH THE UTILIZATION OF THE PR2 GENE - Kamran Ali; Shazia Anwer Bukhari; Bushra Sadia; Mahmood-ur-Rahman (Journal of Animal and Plant Sciences, 2026)
Wheat (Triticum aestivum L.) productivity is severely constrained by biotic stresses, among which stem rust remains a major threat to global food security. In this study, we developed wheat line overexpressing the pathogenesis-related gene 2 (PR2) gene to build innate antifungal defenses. The PR gene coding sequence was cloned into the pCB-PR2-P2A-GUS expression vector and introduced into embryogenic calli of the TD-1 cultivar via biolistic transformation. Regenerated plantlets were subjected to rigorous molecular analysis, including PCR and RT qPCR, confirming stable genomic integration and ~35-fold upregulation of PR2 gene transcripts relative to non-transgenic controls. Histochemical β-glucuronidase (GUS) assays revealed spatial and temporal expression patterns across callus and developing seeds, indicating robust promoter-driven activity. Functional evaluation under natural pathogen exposure demonstrated marked reduced stem rust pustule formation in transgenic lines, validating the antifungal efficacy of β-1,3-glucanase in plants. In silico allergenicity analysis showed minimal similarity to known wheat allergens, supporting the biosafety of the transgene. These results established a comprehensive framework for PR2 gene-mediated disease resistance, integrating molecular validation with phenotypic assessment. Future field trials will elucidate the durability of stem rust resistance and yield stability, providing strategic insights for wheat breeding programs.
Keywords: journal article; Stem resistance; PR2 gene; Transgenic wheat; Wheat transformation