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Abstract

Abstract The invasive weed Parthenium hysterophorus exhibits remarkable adaptability to abiotic stresses, yet the molecular mechanisms underpinning its cold tolerance remain unexplored. This study investigate the role of the DREB1/CBF transcription factor in mediating low-temperature resilience in P. hysterophorus and functional analysis of the DREB1/CBF gene, a key player in cold stress responses. Physiological and molecular analyses were performed on plants exposed to cold (4°C) and control (25°C) conditions. Physiological assays revealed cold-acclimated plants (4°C) exhibited enhanced membrane stability (21% electrolyte leakage) compared to non-acclimated controls (48%), alongside a 19.9% increase in carotenoid content, suggesting antioxidant compensation for oxidative stress mitigation. A notable decrease found in chlorophyll a (2.78 µg/g) and chlorophyll b (1.26 µg/g) in cold-treated plants compared to controls (3.14 µg/g and 1.62 µg/g, respectively). Molecular analysis identified a novel DREB1/CBF homolog, showing significant up-regulation under cold stress via PCR amplification. Bioinformatics revealed >90% nucleotide sequence homology with Brassica species, with conserved AP2/ERF DNA-binding domains critical for CRT/DRE motif recognition. Structural modeling predicted a stable globular protein with solvent-exposed β-sheets, validated through refinement (ERRAT: 95.6%; Ramachandran favored regions: 93.3%). Phylogenetic clustering with Brassica napus and B. oleracea underscores evolutionary conservation of cold-response pathways. These findings provide novel insights into the molecular basis of invasiveness in P. hysterophorus, highlighting dual applications for targeted weed management and the development of climate-resilient crops through DREB1/CBF engineering.

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