# Establishing Reagent Testing Platforms for Functional Analyses in Sunflower

## TL;DR

This paper develops and compares methods for delivering genetic reagents in sunflower to enable functional testing and genetic engineering.

## Contribution

The study establishes scalable and efficient reagent delivery platforms for functional analyses in sunflower.

## Key findings

- Three reagent delivery methods were successfully adapted for sunflower seedlings.
- Agrobacterium-mediated tissue co-culture showed varying effectiveness with different strains and reporter genes.
- The platforms provide a foundation for advanced genetic engineering in sunflower.

## Abstract

Recent advancements in molecular tools for plant genetic engineering, particularly CRISPR-based technologies, have created new opportunities for targeted genome editing. However, applying these tools remains challenging in crop species such as sunflower (Helianthus annuus) that lack established and effective transformation pipelines, including transient reagent delivery methods for functional screening and validation of genetic engineering tools. To address this gap, three major reagent delivery platforms, namely protoplast transfection, leaf infiltration, and Agrobacterium-mediated tissue co-culture, were systematically adapted and assessed for use in sunflower seedlings. While each method enabled successful reagent delivery, they differed in their levels of scalability and efficiency. With these platforms, delivery by different Agrobacterium strains and the effectiveness of various reporter gene expression cassettes were compared to define the most experimentally suitable components for different applications in sunflowers. Together, these results establish a foundational toolkit for transient functional testing in sunflower and pave the way for more sophisticated genetic engineering approaches in this agriculturally important oilseed, confectionary seed, and horticultural crop.