Evidence for nanoparticle-enhanced ultrasound-based In-Planta transformation and transgenerational inheritance in wheat

Authors: RAFIEI, FARIBA - Purdue University; NEPAL, NIRMAN - Purdue University; THURGOOD, TREVER - Purdue University; WILHELM, ROLAND - Purdue University; Scofield, Steven; MOHAMMADI, MOHSEN - Purdue University

Submitted to: New Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/9/2025
Publication Date: 3/25/2026
Citation: Rafiei, F., Nepal, N., Thurgood, T., Wilhelm, R., Scofield, S.R., Mohammadi, M. 2026. Evidence for nanoparticle-enhanced ultrasound-based In-Planta transformation and transgenerational inheritance in wheat. New Biotechnology. https://doi.org/10.1016/j.nbt.2025.12.003.
DOI: https://doi.org/10.1016/j.nbt.2025.12.003

Interpretive Summary: Wheat is ranks as the second most produced cereal for human consumption, however wheat improvement has been impeded by the inability to apply biotechnology traits. The main block to this is the inability to introduce DNA constructs into the plant. In this work we describe development of a simple and efficient system for wheat transformation that overcomes this inability. It does not involve tissue culture and perhaps most importantly is genotype independent unlike any previous wheat transformation methods. This system has the potential to make possible editing genes to improve disease resistance, remove glutens from seed proteins, or creating dwarfing phenotypes that were critical for the green revolution. Because of this method's genotype-independence, biotech improvement could be done directly in the elite wheat variety that farmers are currently growing or that seed companies are developing for future sale, thereby greatly reducing the time to market for US farmers.. This breakthrough is a gateway for applying biotechnology for wheat improvement.

Technical Abstract: Here, we describe an ultrasound-mediated, nanoparticle-based gene delivery system to 12 introduce DNA into cells of mature wheat seeds, which produce transformed T0 plants, without 13 the need for tissue culture. This approach offers a potentially high-throughput, amenable to 14 automation, and a cost-effective strategy for transforming wheat seeds and stably transmitting 15 transgenes across generations, with stable inheritance confirmed through the T3 generation. The 16 method successfully delivered transgenes, via the pCAMBIA1305.1 plasmid, to multiple wheat 17 varieties, including ‘Apogee’, ‘Sonora-64’ and ‘Opata-85’, indicating its potential as a genotype-18 independent platform for plant genetic engineering. Ultrasound treatment transiently increases 19 the permeability of seed cell walls and membranes, facilitating the uptake of mesoporous silica 20 nanoparticles coated with polyethyleneimine that are loaded with plasmid DNA. Quantitative 21 PCR, segregation analysis, and whole-genome sequencing confirmed stable integration of 22 segments of plasmid DNA across multiple loci. Transgene expression was validated using GUS 23 staining and PCR assays across successive generations, supporting robust transgenerational 24 inheritance. By eliminating the need for callus induction or genotype-specific regeneration 25 protocols, this genotype3-independent method of direct seed-transformation offers a potentially 26 game-changing approach for production large numbers of stably transformed wheat plants with 27 demonstrated heritable transmission of transgene expression.