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Abstract:
Bacterial infection affects plant host’s gene expression by activating the innate immune system and signaling cascades, like phytohormone pathways. In plant infection biology several advances have been made by identifying bacterial taxa from natural populations and studying plant immune system. However, we lack understanding on how the bacterial quantity and diversity in the leaf affect the host tissues’ spatial gene expression. To fill this gap, we applied an innovative, high-throughput technology originally developed for mammalian tissues, Spatial Transcriptomics, which enables the simultaneous quantification and visualization of transcriptional profiles in tissue sections at 55-μm resolution, to outdoor-grown Arabidopsis thaliana leaves. We developed several advancements to the original method in order to study the concerted bacterial infection process and plant response in Arabidopsis thaliana leaves. First, we demonstrated with Pseudomonas infected leaves that we are able to capture the bacterial RNA while preserving the complete morphology of the plant tissue. Second, we showed that the tissue treatments we introduced allow us to also spatially capture the plant mRNAs. Third, we demonstrated the capture of bacterial diversity from outdoor grown A. thaliana leaves. In conclusion, our results indicate the feasibility of studying combined gene expression profiles of the host plant and the natural microbial diversity opening up the possibility of extending our approach to different plant systems such as crop species to elucidate complex infection processes where the spatial component is key for their understanding.
