Abstract
Globally is a rising rate of antimicrobial resistance (AMR) that is posing an urgent demand on the need to find safe, sustainable, and effective alternative to traditional antibiotics. This paper investigated the antimicrobial promise of plant-derived cyclotides and green-synthesized metal oxide nanoparticles and combines the fields of molecular biology, phytochemistry, and Nano biotechnology. Cyclotide mining in petunia species (Solanaceae) and Morus nigra leaf extract in the synthesis of copper oxide (CuO) nanoparticles in an environmentally friendly manner were chosen. Polyphenol-rich tissues were optimally used in the extraction of high-quality genomic DNA by a CTAB protocol that has been modified and allowed a successful amplification of the cyclotide genes through PCR. The expression of the protein by recombinant expression confirmed the presence of functional cyclotide precursors by cloning to pJET and pATX-SUMO vectors and recombinant protein production was confirmed by SDS-PAGE analysis. CuO nanoparticles were synthesized greenly by the reduction of CuSO 4 with the help of Morus nigra extract which was demonstrated by the color change and long-term stability of the nanoparticles. Both crude plant extracts and synthesized nanoparticles showed a high inhibitory activity in antibacterial and antifungal against Escherichia coli, Staphylococcus aureus, Aspergillus flavus, and Fusarium oxysporum. It is also important to note that the joint effects of cyclotide-rich extracts and CuO nanoparticles gave a greater inhibition zone, which implies a synergistic effect. Insecticidal bioassays also indicated that there was a high larval mortality in Helicoverpa armigera which confirmed the broad spectrum bioactivity of the agents used. The results show that plant-derived cyclotides and green-synthesized nanoparticles represent the perspective antimicrobial and agricultural agents. The study helps to create a solid foundation on which in vivo research can be performed in the future and solutions based on nature can be developed to fight AMR and enhance crop protection.