2024/04/16 by Daisong Pan, José A. Rivera, Max Miao +10 · 1 voice
Agricultural and Biological Sciences · Biochemistry, Genetics and Molecular Biology · #Advanced Fluorescence Microscopy Techniques #Fern and Epiphyte Biology #Plant Molecular Biology Research
paper · pdf · doi:10.1101/2024.04.13.589377
openalex publication_date 2024/04/16 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/04
ABSTRACT Root biology is pivotal in addressing global challenges including sustainable agriculture and climate change. However, roots have been relatively understudied among plant organs, partly due to the difficulties in imaging root structures in their natural environment. Here we used microfabricated ecosystems (EcoFABs) to establish growing environments with optical access and employed nonlinear multimodal microscopy of third-harmonic generation (THG) and three-photon fluorescence (3PF) to achieve label-free, in situ imaging of live roots and microbes at high spatiotemporal resolution. THG enabled us to observe key plant root structures including the vasculature, Casparian strips, dividing meristematic cells, and root cap cells, as well as subcellular features including nuclear envelopes, nucleoli, starch granules, and putative stress granules. THG from the cell walls of bacteria and fungi also provides label-free contrast for visualizing these microbes in the root rhizosphere. With simultaneously recorded 3PF fluorescence signal, we demonstrated our ability to investigate root-microbe interactions by achieving single-bacterium tracking and subcellular imaging of fungal spores and hyphae in the rhizosphere.