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Abstract

The invasive weed Parthenium hysterophorus exhibits remarkable adaptability to abiotic stresses, yet the molecular mechanisms underpinning its cold tolerance remain unexplored. This study investigates the role of the DREB1/CBF transcription factor in mediating low-temperature resilience in P. hysterophorus and functional analysis of the DREB1/CBF gene. Physiological and molecular analyses were performed on plants exposed to cold (4°C) and control (25°C) conditions. Cold-acclimated plants (4°C) exhibited enhanced membrane stability compared to non-acclimated control plants, Carotenoids contents increased 19.9% in cold acclimated plants compared to control. 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 first ever novel DREB1/CBF homolog in P. hysterophorus, 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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