Sustainability in developing novel nanomaterials (NPs) from biomass sources is a challenging proposition mainly due to the difficulty of infusing or retaining desired chemical functionalities in the biomass substrate. In this study, we demonstrate the synthesis of DNA-nanoparticles (DNA-NP) from onion genomic DNA as a plant biomass source through controlled hydrothermal pyrolysis to retain functional groups in the NPs for predictable downstream chemical transformations. A dual crosslinking scheme was introduced that involves the DNA-NP to form a hydrogel. Chemical crosslinking was achieved through the formation of a Schiff base between the –CHO groups of glutaraldehyde and the amine functionality present on the DNA-NP surface as well as in the nucleobases of the dangling DNA strands of DNA-NP. Simultaneous physical entanglement was attained through hybridization-mediated self-assembly of the dangling DNA strands of the DNA-NP with untransformed onion genomic DNA. As a corollary of the dual crosslinking, the resulting hydrogel not only displayed remarkable mechanical strength but also showed self-healing properties. The ability of the DNA-NP to generate reactive oxygen species (ROS) with visible light irradiation is translated to the hydrogel, making the system potent for biofilm destruction. The high loading efficiency of the model drug ampicillin sodium (Amp) in the hydrogel was achieved which was released in four days. This hints towards the application of the hydrogel through combination antibiotic–antibacterial photodynamic treatment (APDT) as demonstrated here with both Gram-positive and Gram-negative bacteria.