Time-Lapse Imaging of Root Pathogenesis and Fungal Proliferation Without Physically Disrupting Roots

Abstract

Microscopic observation of root disease onset and progression is typically performed by harvesting different plants at multiple time points. This approach prevents the monitoring of individual encounter sites over time, often mechanically damages roots, and exposes roots to unnatural conditions during observation. Here, we describe a method developed to avoid these problems and its application to study Fusarium oxysporum-Arabidopsis thaliana interactions. This method enabled three-dimensional, time-lapse imaging of both A. thaliana and F. oxysporum as they interact via the use of confocal and multi-photon microscopy and facilitated inquiries about the genetic mechanism underpinning Fusarium wilt.

Keywords: Arabidopsis thaliana; Calcium indicator; Confocal microscopy; Fluorescent protein; Fusarium oxysporum; Root colonization; Soilborne disease.