# Hypericin-Mediated Photodynamic Inactivation Against the Plant Pathogen Clavibacter michiganensis: Preventative Seed Decontamination Enhanced by Potassium Iodide

## TL;DR

This study shows that a hypericin-based treatment combined with light and potassium iodide can effectively kill a tomato seed-borne bacteria without harming seed germination.

## Contribution

A novel hypericin-polyvinylpyrrolidone complex with potassium iodide is shown to effectively decontaminate tomato seeds from Clavibacter michiganensis.

## Key findings

- HHL-PVP with red light achieves >99.9% bacterial reduction in liquid culture.
- Combining HHL-PVP with potassium iodide completely eradicates C. michiganensis in liquid and on seeds.
- Seed decontamination with HHL-PVP and KI maintains sprouting rates while killing all bacteria.

## Abstract

Clavibacter michiganensis causes significant crop losses in tomatoes, and the disease may be transferred by plant seeds. This study evaluates the efficacy of Photodynamic Inactivation (PDI) with a water-soluble hypericin derivative, developed as a complex with polyvinylpyrrolidone (high hypericin-loaded PVP, HHL-PVP), as a decontamination strategy for tomato seeds. HHL-PVP was chosen for its overall stability, as the complex remains stable in solution for over 950 days, maintains its absorption capacity after illumination with 200 J·cm−2, and produces reactive oxygen species (ROS) even at concentrations as low as 1 µM. PDI against C. michiganensis with 5 μM HHL-PVP, 10 min drug to light interval (DLI), and illumination with red light (600–700 nm, 100 J·cm−2) exceeded the antimicrobial effect of a 99.9% reduction in liquid culture. Increasing the DLI to 24 h did not alter the photokilling effect. A 14 h light/10 h dark cycle in white light (118 J·cm−2) with 0.3 µM HHL-PVP inhibited the growth of C. michiganensis by more than 6 log10 steps, indicating that HHL-PVP has a stable and long-lasting photokilling effect. The combination of HHL-PVP with potassium iodide (KI, 100 mM) completely eradicated C. michiganensis in liquid culture with red and white light, indicating KI’s role in enhancing phototoxicity. Tomato seed photodynamic decontamination using 1.0 µM HHL-PVP activated by 200 J·cm−2 white light inactivated >5 log10 of C. michiganensis, without diminishing sprouting. An addition of 100 mM KI increased the percentage of sprouted seedlings and inactivated 100% of bacteria. These results demonstrate that HHL-PVP-mediated PDI combined with KI could be highly effective as a preventative strategy in tomato protection against C. michiganensis.