Bio-engineered As-Ag-TiO2 nanoparticles enhance the genetic transformation of Pisum sativum L. via proton-coupled electron transfer-dependent alternative protonation

• Published in: Industrial crops and products 2024-09, Vol.215, p.118604, Article 118604
• Main Authors: Chandrasekaran, Ajithan, Pilavadi, Thangamuniyandi, Venkatachalam, Vasudevan, Umapathy, Devan, Arockiam, Antony Joseph Velanganni, Singarayar, Magdalin Sylvia, Lee, Geung-Joo, Markandan, Manickavasagam
• Format: Article
• Language: English
• Subjects: Acetosyringone; Advanced gene transformation; Bio-engineered nanoparticles; Nano-doping; PCET; Pea herbicide resistance
• ISSN: 0926-6690; 1872-633X
• DOI: 10.1016/j.indcrop.2024.118604

Nanoparticle-assisted biomolecule delivery to improve plant gene transformation and expression is a, cutting-edge approach. In this study, we examined the effects of acetosyringone-doped BENPs on the enhanced genetic transformation of peas. The vir gene inducer acetosyringone was delivered into Agrobacterium via a nano-doping system with TiO2 and Ag NPs. Four BENPs, such as TiO2 (S1), Ag-TiO2 (S2), As-TiO2 (S3), and As-Ag-TiO2 (S4), were green-synthesized from the PLE using hydrothermal and calcination approaches. SEM, HR-TEM-EDAX, SAED, XRD, and XPS physicochemical characterizations ensured the BENPs structure and biomolecule doping. The potential bar transformants were chosen against the MIC of BASTA® using the EHA 105 Agrobacterium tumefaciens strain carrying the pCAMBIA 1304-plasmid. The delivery of nano-doped acetosyringone was studied with various applications, and the typical exogenous acetosyringone delivery in the absence of BENPs was also analyzed as a control experiment. The gus histochemical, PCR, and southern hybridization confirmed that, among the BENPs, S4 at a 4 ppm dose induced the highest gene transformation efficiency (36.33 %) in pea along with three days of co-cultivation, 30 sec of sonication, and 3 min of vacuum infiltration. In BENP-assisted pea gene transformation, S2 at 4 ppm (26.66 %), S3 at 3 ppm (23 %), and S1 at 4 ppm (17.66 %) showed the next three significant gene transformation efficiencies, respectively. The elevated virC and virG gene expression profiles of the S4 BENP-treated Agrobacterium were identified with gene-specific RT-PCR analysis. The PCET activity of S4 was critically confirmed with cyclic voltammetry, which revealed that BENPs might function as an alternative proton supplement system to improve the vir gene induction and transformation of T-DNA. Hence, this alternative PCET-dependent BENP-assisted acetosyringone delivery for Agrobacterium mediated gene transformation exhibits potential value in the development of genetically-modified peas and opens the door to crops for advanced genetic manipulation against various biotic and abiotic stresses. [Display omitted] •BENPs were synthesized from pea leaf extract using acetosyringone-doped titanium dioxide (TiO2) and silver (Ag) nanoparticles (As-Ag-TiO2), and the BENPs were characterized by SEM, TEM-EDAX, SEAD, and XPS analysis.•Cyclic voltammetry confirmed proton coupled electron transfer (PCET) activity in BENPs.•The standardized BENPs-assisted advanced pea gene