Biology subjects

Caillaud, M.-C.

Publications and source records attributed to Caillaud, M.-C..

9 recordsLinked to original sources

bioRxiv2026

A triple fluorescent marker for live imaging of plant cell morphogenesis

Live imaging of plant subcellular structures is key to deciphering the spatiotemporal bases of cellular processes, and their functional impact on growth and morphogenesis at various biological scales. Live imaging of plant cells essentially relies on expression of fluorescent markers labeling cells or subcellular structures of interest. Simultaneous multi-channel imaging of several markers is still not routine practice in plant cell biology, owing to issues linked to genetic or spectral compatibility of markers, differences in expression levels, silencing, toxicity, etc. Here we designed a three-color marker in Arabidopsis thaliana and Capsella rubella, enabling high-resolution live imaging of plant morphogenesis, including labeling of the cell membrane, the nucleus and the microtubule cytoskeleton. Detection of MT arrays involved the development of a MAP4-MBD-based microtubule marker optimized for plant cells. The three-color marker allows visualization of the three-dimensional organization and dynamics of plant microtubules within the intracellular space with unprecedented precision, in various organs including the root and shoot meristems, the leaf, anther, and gynoecium. Our results demonstrate the potential of such single-construct strategy for cell biology studies in plants.

plant biology↗

bioRxiv2025

Actin Counters Geometry to Guide Plant Cell Division

In multicellular organisms, cell division shapes tissue architecture, cell identity, and function. In walled organisms like plants, division plane orientation irreversibly defines tissue topology and is tightly regulated. While divisions often follow the shortest path enclosing cell volume, certain cells deviate, dividing perpendicular to the growth axis. Tissue-scale mechanical stress has been proposed to guide such orientation, but how these cues are integrated remains unclear. Here, we re-examine the role of the actin cytoskeleton in orienting cell division in Arabidopsis root epidermis. Combining cell biology, genetics, pharmacological treatments, 3D segmentation, and modelling, we show that actin is a central molecular actor required to establish cell division orientation against the geometrical rules, highlighting its role in integrating spatial information.

plant biology↗

bioRxiv2022

The phosphoinositide signature guides the final step of plant cytokinesis

Plant cytokinesis, which fundamentally differs from that in animals, involves de novo assembly of a plasma membrane precursor named the cell plate. How the transition from the cell plate to a plasma membrane occurs at the end of the plant cytokinesis remains poorly understood. Here, we describe with unprecedented spatiotemporal precision, the acquisition of plasma membrane identity upon cytokinesis through the lateral patterning of phosphatidylinositol 4,5-bisphosphate PI(4,5)P2 at the newly formed cell plate membrane. We show that during late cytokinesis, opposing polarity domains are formed along the cell plate. Exclusion of PI(4,5)P2 from the leading edge of the cell plate is controlled by SAC9, a putative phosphoinositide phosphatase. SAC9 colocalizes with MAP65-3, a key regulator of the cytokinesis, at the cell plate leading zone and regulates its function. In the sac9-3 mutant, the polar distribution of PI(4,5)P2 at the cell plate is altered, leading to de-novo recruitment of the cytokinesis apparatus and to formation of an additional, ectopic cell plate insertion site. We proposed that PI(4,5)P2 acts as a polar cue to spatially separate the expansion and maturation domains of the forming cell plate during the final steps of cytokinesis. One Sentence SummaryThe phosphoinositide PI(4,5)P2 acts as an hallmark to guide the final step of plant cell division.

cell biology↗

bioRxiv2021

The Arabidopsis SAC9 Enzyme defines a cortical population of early endosomes and restricts PI(4,5)P2 to the Plasma Membrane

Membrane lipids, and especially phosphoinositides, are differentially enriched within the eukaryotic endomembrane system. This generates a landmark code by modulating the properties of each membrane. Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] specifically accumulates at the plasma membrane in yeast, animal and plant cells, where it regulates a wide range of cellular processes including endocytic trafficking. However, the functional consequences of mispatterning PI(4,5)P2 in plants are unknown. Here, we functionally characterized the putative phosphoinositide phosphatase SUPPRESSOR OF ACTIN9 (SAC9) in Arabidopsis thaliana (Arabidopsis). We found that SAC9 depletion led to the ectopic localization of PI(4,5)P2 on cortical intracellular compartments, which depends on PI4P and PI(4,5)P2 production at the plasma membrane. SAC9 localizes to a subpopulation of trans-Golgi Network/early endosomes that are enriched in a region close to the cell cortex and that are coated with clathrin. Furthermore, it interacts and colocalizes with Src Homology 3 Domain Protein 2 (SH3P2), a protein involved in endocytic trafficking. In the absence of SAC9, SH3P2 localization is altered and the clathrin-mediated endocytosis rate is reduced. Together, our results highlight the importance of restricting PI(4,5)P2 at the plasma membrane and illustrate that one of the consequence of PI(4,5)P2 misspatterning in plants is to impact the endocytic trafficking. One-sentence summarySAC9 prevents the accumulation of PI(4,5)P2 in intracellular compartments, thereby contributing to clathrin-mediated endocytosis at the plasma membrane and regulating the cortical localization of its protein partner SH3P2.

plant biology↗

bioRxiv2021

Dynamic Apical-Basal Enrichment of the F-Actin during Cytokinesis in Arabidopsis Cells Embedded in Their Tissues

During the life cycle of any multicellular organism, cell division contributes to the proliferation of the cell in the tissues as well as the generation of specialized cells, both necessary to form a functional organism. Therefore, the mechanisms of cell division need to be tightly regulated, as malfunctions in their control can lead to tumor formation or developmental defects. This is particularly true in land plants, where cells cannot relocate and therefore cytokinesis is key for morphogenesis. In the green lineage, cell division is executed in radically different manners than animals, with the appearance of new structures (the preprophase band (PPB), cytokinetic the cell plate and phragmoplast), and the disappearance of ancestral mechanisms (cleavage, centrosomes). While F-actin and microtubules closely co-exist to allow the orientation and the progression of the plant cell division, recent studies mainly focused on the microtubules involvement in this key process. Here, we used our recently developed root tracking system to follow actin dynamics in dividing Arabidopsis meristematic root cells. In this study, we imaged in time and space the fluorescent-tagged F-actin reporter Lifeact together with cell division markers in dividing cells embedded in their tissues. In addition to the F-actin accumulations in the phragmoplasts, we observed and quantified a dynamic apical-basal enrichment of the F-actin during cytokinesis. The role and the possible actors responsible for F-actin dynamics during cytokinesis are discussed.

cell biology↗

bioRxiv2020

A nanodomain anchored-scaffolding complex is required for PI4K? function and localization in plants

Phosphoinositides are low-abundant lipids that participate in the acquisition of membrane identity through their spatiotemporal enrichment in specific compartments. PI4P accumulates at the plant plasma membrane driving its high electrostatic potential, and thereby facilitating interactions with polybasic regions of proteins. PI4K1 has been suggested to produce PI4P at the plasma membrane, but how it is recruited to this compartment is unknown. Here, we pin-point the mechanism that tethers PI4K1 to the plasma membrane via a nanodomain-anchored scaffolding complex. We identified that PI4K1 is part of a complex composed of proteins from the NO-POLLEN-GERMINATION, EFR3-OF-PLANTS, and HYCCIN-CONTAINING families. Comprehensive knock-out and knock-down strategies revealed that subunits of the PI4K1 complex are essential for pollen, embryonic and post-embryonic development. We further found that the PI4K1 complex is immobilized in plasma membrane nanodomains. Using synthetic mis-targeting strategies, we demonstrate that a combination of lipid anchoring and scaffolding localizes PI4K1 to the plasma membrane, which is essential for its function. Together, this work opens new perspectives on the mechanisms and function of plasma membrane nanopatterning by lipid kinases.

plant biology↗

bioRxiv2020

Coiled-coil and RPW8-type immune receptors function at the plasma membrane in a phospholipid dependent manner

Activation of intracellular nucleotide-binding leucine-rich repeat receptors (NLRs) results in immunity and a localized cell death response of infected cells. Cell death activity of many NLRs requires oligomerization and in some cases plasma membrane (PM) localization. However, the exact mechanisms underlying PM localization of NLRs lacking recognizable N- or C-terminal lipidation motifs or predicted transmembrane domains remains elusive. Here we show that the PM localization and stability of members of the RPW8-like coiled-coil (CCR) domain NLRs (RNLs) and a CC-type NLR (CNL) depend on the interaction with PM phospholipids. Depletion of phosphatidylinositol-4-phosphate (PI4P) from the PM led to a mislocalization of the analyzed NLRs and consequently inhibited their cell death activity. We further demonstrate activation-dependent self-association of cell death inducing RNLs. Our results provide new insights into the molecular mechanism of NLR PM localization and defines an important role of phospholipids for CNL and RNL activity during immunity.

plant biology↗

bioRxiv2020

iDePP: a genetically encoded system for the inducible depletion of PI(4,5)P2 in Arabidopsis thaliana.

Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] is a low abundant lipid present at the plasma membrane of eukaryotic cells. Extensive studies in animal cells revealed the pleiotropic functions of PI(4,5)P2. In plant cells, PI(4,5)P2 is involved in various cellular processes including the regulation of cell polarity and tip growth, clathrin-mediated endocytosis, polar auxin transport, actin dynamics or membrane-contact sites. To date, most studies investigating the role of PI(4,5)P2 in plants have relied on mutants lacking enzymes responsible for PI(4,5)P2 synthesis and degradation. However, such genetic perturbations only allow steady-state analysis of plants undergoing their life cycle in PI(4,5)P2 deficient conditions and the corresponding mutants are likely to induce a range of non-causal (untargeted) effects driven by compensatory mechanisms. In addition, there are no small molecule inhibitors that are available in plants to specifically block the production of this lipid. Thus, there is currently no system to fine tune PI(4,5)P2 content in plant cells. Here we report a genetically encoded and inducible synthetic system, iDePP (Inducible Depletion of PI(4,5)P2 in Plants), that efficiently removes PI(4,5)P2 from the plasma membrane in different organs of Arabidopsis thaliana, including root meristem, root hair and shoot apical meristem. We show that iDePP allows the inducible depletion of PI(4,5)P2 in less than three hours. Using this strategy, we reveal that PI(4,5)P2 is critical for cortical microtubule organization. Together, we propose that iDePP is a simple and efficient genetic tool to test the importance of PI(4,5)P2 in given cellular or developmental responses but also to evaluate the importance of this lipid in protein localization. Research OrganismA. thaliana

plant biology↗

bioRxiv2018

A combinatorial lipid code shapes the electrostatic landscape of plant endomembranes

Membrane surface charge is critical for the transient, yet specific recruitment of proteins with polybasic regions to certain organelles. In all eukaryotes, the plasma membrane (PM) is the most electronegative compartment of the cell, which specifies its identity. As such, membrane electrostatics is a central parameter in signaling, intracellular trafficking and polarity. Here, we explore which are the lipids that control membrane electrostatics using plants as a model. We show that phosphatidic acidic (PA), phosphatidylserine (PS) and phosphatidylinositol-4-phosphate (PI4P) are separately required to generate the electrostatic signature of the plant PM. In addition, we reveal the existence of an electrostatic territory that is organized as a gradient along the endocytic pathway and is controlled by PS/PI4P combination. Altogether, we propose that combinatorial lipid composition of the cytosolic leaflet of cellular organelles not only defines the plant electrostatic territory but also distinguishes different compartments within this territory by specifying their varying surface charges.

plant biology↗