Fuente: PubMed "industrial biotechnology" 3 Biotech. 2026 Aug;16(8):318. doi: 10.1007/s13205-026-04963-0. Epub 2026 Jul 9.ABSTRACTA direct organogenesis-mediated Agrobacterium tumefaciens-based transformation system was developed in pea (Pisum sativum L. cv. Ageta 6) using green-synthesized bioengineered nanoparticles (BENPs) to improve transformation efficiency. Titanium dioxide (TiO₂), silver (Ag), and acetosyringone (AS)-doped AS-Ag-TiO₂ BENPs were synthesized through a green hydrothermal approach and evaluated under optimized co-cultivation, sonication, and vacuum infiltration conditions. The synthesized BENPs were previously characterized through XRD, SEM, EDAX, HR-TEM, SAED, XPS, and cyclic voltammetry analyses, confirming ultrafine crystalline nanostructures and stable nanocomposite formation. Regenerated putative transformants were assessed through RAPD-based genetic fidelity analysis and further validated by histochemical GUS assays, bar gene-specific PCR amplification, and RT-PCR expression analysis. Among all treatments evaluated, AS-Ag-TiO₂ BENPs at 4 ppm produced the highest transformation efficiency (30.66%), followed by AS-TiO₂ (23.33%), Ag-TiO₂ (18.33%), and TiO₂ (13.33%), whereas the conventional AS-mediated transformation system produced only 8.66% transformation efficiency under identical optimized conditions and BASTA® selection pressure. RT-PCR analysis further demonstrated relatively enhanced bar transcript accumulation in AS-Ag-TiO₂-treated transformants compared with other BENPs-assisted treatments. The enhanced transformation response may be associated with improved Agrobacterium attachment, vir gene activation, intracellular T-DNA transfer, and enhanced explant survival during co-cultivation. In addition, the AS-Ag-TiO₂ nanocomposite may facilitate proton-coupled electron transfer (PCET)-associated alternative protonation mechanisms supporting enhanced VirA/VirG signalling and transformation competency. Collectively, the present study establishes an efficient and cost-effective BENPs-assisted transformation platform for recalcitrant legumes and highlights the potential application of multifunctional nanocomposites in plant genetic transformation and nano-enabled biomolecule delivery systems.SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-026-04963-0.PMID:42434792 | PMC:PMC13350613 | DOI:10.1007/s13205-026-04963-0