Visualization of root extracellular traps in an ectomycorrhizal woody plant (Pinus densiflora) and their interactions with root-associated bacteria

Shirakawa, Makoto; Norihisa Matsushita; Kenji Fukuda

doi:10.51094/jxiv.333

##article.authors##

Shirakawa, Makoto Graduate School of Agricultural and Life Sciences, The University of Tokyo https://orcid.org/0000-0003-3454-6768
Norihisa Matsushita Graduate School of Agricultural and Life Sciences, The University of Tokyo
Kenji Fukuda Graduate School of Agricultural and Life Sciences, The University of Tokyo

DOI:

https://doi.org/10.51094/jxiv.333

キーワード:

Border cells、 Japanese red pine、 Paraburkholderia、 Primary root、 Rhizobacteria、 Root mucilage

抄録

Ectomycorrhizal (ECM) woody plants, such as members of Pinaceae and Fagaceae, can acquire resistance to biotic and abiotic stresses through the formation of mycorrhiza with ECM fungi. However, germinated tree seedlings do not have mycorrhizae and it takes several weeks for ectomycorrhizae to form on their root tips. Therefore, to confer protection during the early growth stage, bare primary roots require defense mechanisms other than mycorrhization. Here, we attempted to visualize root extracellular traps (RETs), an innate root defense mechanism, in the primary root of Pinus densiflora and investigate the interactions with root-associated bacteria isolated from ECM and fine non-mycorrhizal roots. Histological and histochemical imaging and colony forming unit assays demonstrated that RETs in P. densiflora, mainly consisting of root-associated, cap-derived cells (AC-DCs) and large amounts of root mucilage, promoted bacterial colonization in the rhizosphere, despite also having bactericidal activity via extracellular DNA. Four rhizobacterial strains induced the production of reactive oxygen species (ROS) from host tree AC-DCs without being excluded from the rhizosphere of P. densiflora. In particular, applying two Paraburkholderia strains, PM O-EM8 and PF T-NM22, showed significant differences in the ROS levels from the control group. These results reveal an indirect contribution of rhizobacteria to host root defense, and suggest that root-associated bacteria could be a component of RETs as a first line of defense against root pathogens in the early growth stage of ECM woody plants.

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