Augmentation of betacyanin and quercetin in hybrid callus: Comprehensive assessment of biosynthesized silver nanoparticles for their potent biological activities, advanced in silico interactions, and rigorous toxicological evaluation

The growing demand for natural compounds with potent biological activities has prompted extensive research into alternative sources of antioxidants and antibacterial agents. This study discusses a new approach to producing a fused hybrid protoplast using Cyperus pangorei Rottb (Korai), and Amaranthus dubius Mart. ex Thell (Red spinach) . The screening of hybrid protoplasts and induction of hybrid callus were attained by Murashige and Skoog (MS) medium with 1.5 ppm/L benzylaminopurine (BAP) and 0.5 ppm/L 2,4-dichlorophenoxyacetic acid (2,4-D) under a 16-h light photoperiod. The fused hybrid protoplast, via microcalli formation, enhanced betacyanin and quercetin with the influence of different elicitors. The betacyanin (33.36 ± 0.3 µg/mL) and quercetin (24.06 ± 0.3 µg/mL) compounds were recognized and quantified using high-performance liquid chromatography. The hybrid callus-derived AgNPs exhibited a unique 279 nm absorption peak, supported by XRD, EDX, FT-IR, and zeta potential analysis. The methanol extract of hybrid callus (MEHC) demonstrated strong antibacterial activity against Klebsiella pneumoniae (11.30 ± 0.2 mm) and potent free-radical scavenging (DPPH (2, 2-diphenly-1-picrlyhydrazyl): 11.84 ± 0.8; ABTS (2, 2′-azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid): 11.84 ± 0.8).The MEHC toxicity analysis on zebrafish showed a mortality percentage of 0.3–0.2 eggs at 96 h. In a molecular docking study, betacyanin and quercetin showed the best binding affinity (−9.4 and −8.8 kcal/mol) against the 3HR4 target protein. In conclusion, this approach emphasizes the critical role of the modern biotechnological world in maximizing the yield and efficacy of bioactive compounds derived from plants. • The production of fused hybrid protoplast from the Cyperus pangorei and Amaranthus dubius . • The enhanced levels of betacyanin and quercetin from the fused hybrid callus. • The synthesis and characterization of silver nanoparticles and its biological and toxicity effects.