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Pluis, M. H.

Publications and source records attributed to Pluis, M. H..

2 recordsLinked to original sources

Deep learning enables quantitative subcellular analysis of plant-microbe interfaces

Specialized host-microbe interfaces are central to cellular interactions in plants. Intracellular structures such as haustoria formed by filamentous pathogens mediate nutrient exchange and effector delivery to host cells. Despite their biological importance, the lack of quantitative frameworks has largely confined the study of these interfaces to qualitative observations, limiting our ability to compare infection strategies, cellular responses, and spatial organization across cells and tissues. Here, we present HFinder, a deep learning-based framework for automated detection, segmentation, and quantitative analysis of plant-microbe interfaces in confocal images. Using an object-centric deep learning approach, HFinder enables robust identification of haustoria, microbial hyphae, and host organelles across diverse imaging conditions and pathosystems. We demonstrate that this framework supports quantitative analyses of subcellular processes at host-microbe interfaces, including effector secretion, perturbation of host cellular processes, and immune receptor accumulation at haustoria. HFinder provides a practical and scalable solution for the systematic digitalization of plant infection imaging data and establishes a general framework for quantitative studies of cellular dynamics at host-microbe contact zones.

cell biology↗

LbCas12-mediated multiplex gene editing and 2-fluoroadenine counter-selection in Phytophthora palmivora

CRISPR-Cas systems have moved forward genetic engineering in virtually any organism amenable to genetic modification. In particular, these systems have unlocked unprecedented possibilities to generate mutants in oomycetes, a group of filamentous microbes comprising over two hundred Phytophthora species, including the cacao killer Phytophthora palmivora. Here, we showcase multiplex gene editing in P. palmivora using LbCas12. We have developed a straightforward protocol to simultaneously knock out two genes encoding adenine phosphoribosyltransferase (APT), an essential enzyme of the purine salvage pathway. We show that APT knockouts ({Delta}PpATP1/2) are insensitive to 2-fluoroadenine (2-FA) and retain full virulence on Nicotiana benthamiana. We rely on zoospore electroporation using an all-in-one construct to facilitate the rapid editing of multiple genes. This work enhances the genetic toolbox for Phytophthora species and simplifies the exploration of gene function, laying the groundwork for future innovations aiming to tackle oomycete plant diseases.

microbiology↗