Effects of Green Synthesized Copper-Derived Nano Particles extracted from Piper betel to Eliminate Contaminations in tissue culture of Dracaena sanderiana Sander ex Mast. cv ‘Gold’

Abstract

Micropropagation is often hindered by contamination from endophytic microbes. While antibiotics are used to eliminate these without harming plant tissues, they risk promoting antibiotic resistance. This study explores a green alternative by synthesizing copper nanoparticles (Cu-NPs) using antimicrobial-rich Piper betel leaf extract. The eco-friendly method uses plant biomolecules as natural reducing and stabilizingagents. The aim was to (i) produce Cu-NPs via eco-friendly plant-based synthesis, (ii) evaluate their antibacterial activity against Escherichia coli and S. aureus, and (iii) assess their effectiveness in reducing microbial contamination in plant tissue culture. Cu-NPs were produced by reacting leaf extract with cupric sulphate under controlled heating and stirring. Their antibacterial activity was tested against E. coli and Staphylococcus aureus via time-kill assays, and their efficacy in reducing contamination was assessed by applying them to Dracaena sanderiana explants in vitro. 200 ppm was identified as the optimal concentration for the antibacterial activity in in vitro establishment trials of Dracaena sanderiana, 50 ppm Cu-NPs proved to be the most effective concentration, resulting in 100% survival when used both for surface sterilization and as a supplement in the culture medium, and 80% survival when used only for surface sterilization. Thefindings suggest that green-synthesised Cu-NPs from P. betel show potential as an alternative to antibiotics in plant tissue culture, with promising effects on contamination control and reduced risk of antibiotic resistance.