Pollen magnetofection platform for tissue culture-free genome editing in white lupin
A nanoparticle-based DNA delivery method that uses magnetic fields to introduce foreign DNA directly into white lupin pollen cells, enabling genome editing without tissue culture. This approach addresses a key bottleneck for Lupinus albus, a recalcitrant species, and offers a faster, cost-effective path to improved cultivars.
Overview
White lupin (Lupinus albus) is a valuable protein crop with a strategic role in crop rotation systems, but it remains difficult to improve through conventional breeding tools because it is recalcitrant to tissue culture. This means that standard genetic transformation and genome editing protocols, which rely on regenerating plants from cultured cells, are not readily available for the species. The proposed solution overcomes this limitation by delivering DNA directly into pollen grains using functionalized magnetic nanoparticles and an external magnetic field, a technique known as pollen magnetofection. By bypassing tissue culture entirely, the approach offers a faster, simpler, and more cost-effective route to genome editing in white lupin and potentially other dicot species.
Technical specifications
- Core technology: Fe3O4 magnetic nanoparticles (MNPs) surface-modified with polyethyleneimine to enable DNA binding and cellular uptake.
- Cargo: Linear DNA constructs, including CRISPR vectors, conjugated to the functionalized MNPs. Conjugation has already been confirmed through Z-potential measurements and TEM analysis.
- Delivery mechanism: An external magnetic field drives the DNA-loaded nanoparticles through the natural pores of pollen grains, transforming pollen cells without physical damage.
- Workflow: Transformed pollen is applied to emasculated lupin flowers, producing transformed seeds without any tissue culture step.
- Validation tools: A plasmid encoding green fluorescent protein (GFP) will be used as a reporter to confirm successful transformation as a proof of concept.
- Supporting expertise: Built on prior work demonstrating magnetofection in cotton, sunflower, and lily, and on validated use of linear DNA in biolistic wheat transformation.
Technology readiness level
The platform is at an early-to-mid development stage. DNA–MNP conjugation has already been achieved and characterized, and the team has prior experience with linear DNA constructs in related transformation systems. The next phase focuses on protocol development for Lupinus albus, including optimization of pollen viability, collection timing, and liquid pollen culture media. Initial proof-of-concept validation will use a GFP reporter before moving to CRISPR-based genome editing applications.
About Instituto de Investigaciones Agropecuarias
INIA is Chile’s national agricultural research and innovation institute, a private non‑profit corporation linked to the Ministry of Agriculture, with roughly 1,000 staff and a network of 11 regional research centers across the country. Co‑located experimental farms and specialized laboratory services enable field‑to‑lab collaboration for partners. A national germplasm bank network and an International Depository Authority microbial collection support characterization, secure deposits, and downstream commercialization. Core funding comes via a performance agreement with the Undersecretariat of Agriculture, complemented by competitive national R&D and innovation programs and revenue from services, seeds, and varieties. Dedicated extension and technology transfer programs accelerate adoption through demonstrations, training, and variety release to producers and agri‑food firms.
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