Influence of Pulsed Electric Field Protocols on the Reversible Permeabilization of Rucola Leaves

Reversible electropermeabilization of plant tissues with heterogeneous structure represents a technological challenge as the response of the different structures within the same specimen to the application of electric field may differ due to different cell sizes, extracellular space configurations, and electrical properties. The influence of five different pulsed electric field protocols with different pulse polarity, number of pulses (25, 50, 75, 100, 250, and 500), and intervals between pulses (no intervals and 1- and 2-ms intervals) on the reversible permeabilization of rucola (Eruca sativa) leaves was investigated. The electric field intensity was 600 V/cm. Electrical resistance of the bulk tissue was measured before and after electroporation, and propidium iodide was used to analyze the electroporation at the surface of the leaf. Leaf viability was assessed from survival in storage, and cell viability was investigated with fluorescein diacetate. Results indicate that the viability of the leaves could not be predicted by measurements of electrical resistance or permeabilization levels of the leaf surface. Higher survival rate was demonstrated when applying bipolar pulses compared with monopolar pulses, but the latter proved to be more effective than bipolar pulses for permeabilizing the surface of the leaves. Longer intervals between bipolar pulses resulted in increased viability preservation, while the number of electroporated cells on the leaf surface was comparable for all tested protocols.