Homologous recombination is central to repair DNA double-strand breaks, either accidently arising in mitotic cells or in a programed manner at meiosis. Crossovers resulting from the repair of meiotic breaks are essential for proper chromosome segregation and increase genetic diversity of the progeny. However, mechanisms regulating crossover formation remain elusive. Here, we identified through genetic and protein-protein interaction screens FIDGETIN-LIKE-1 INTERACTING PROTEIN (FLIP) as a new partner of the previously characterized anti-crossover factor FIDGETIN-LIKE-1 (FIGL1) in Arabidopsis thaliana. We showed that FLIP limits meiotic crossover together with FIGL1. Further, FLIP and FIGL1 form a protein complex conserved from Arabidopsis to human. FIGL1 interacts with the recombinases RAD51 and DMC1, the enzymes that catalyze the DNA strand exchange step of homologous recombination. Arabidopsis flip mutants recapitulate the figl1 phenotype, with enhanced meiotic recombination associated with change in counts of DMC1 and RAD51 foci. Our data thus suggests that FLIP and FIGL1 form a conserved complex that regulates the crucial step of strand invasion in homologous recombination. Homologous recombination is a DNA repair mechanism that is essential to preserve the integrity of genetic information and thus to prevent cancer formation. Homologous recombination is also used during sexual reproduction to generate genetic diversity in the offspring by shuffling parental chromosomes. Here, we identified a novel protein complex (FLIP-FIGL1) that regulates homologous recombination and is conserved from plants to mammals. This suggests the existence of a novel mode of regulation at a central step of homologous recombination.

A major challenge in morphometrics is to analyse complex biological shapes formed by structures at different scales. Leaves exemplify this challenge as they combine differences in their overall shape with smaller shape variations at their margin leading to lobes or teeth. Current methods based on contour or on landmarks analysis are successful in quantifying either overall leaf shape or leaf margin dissection, but fail in combining the two. Here, we present a comprehensive strategy and its associated freely available platform for the quantitative, multiscale analysis of the morphology of leaves with different architectures. For this, biologically relevant landmarks are automatically extracted and hierarchized, and used to guide the reconstruction of accurate average contours that properly represent both global and local features. Using this method we established a quantitative framework of the developmental trajectory of Arabidopsis leaves of different ranks and retraced the origin of leaf heteroblasty. When applied to different mutant forms our method can contribute to a better comprehension of gene function as we show here for the role of CUC2 during Arabidopsis leaf serration. Finally, we illustrated the wider applicability of our tool by analysing hand morphometrics.

Summary:NucleusJ is a simple and user-friendly ImageJ plugin dedicated to the characterization of nuclear morphology and chromatin organization in 3D. Starting from image stacks, the nuclear boundary is delimited by combining the Otsu segmentation method with optimization of nuclear sphericity. Chromatin domains are segmented by partitioning the nucleus using a 3D watershed algorithm and by thresholding a contrast measure over the resulting regions. As output, NucleusJ quantifies 15 parameters including shape and size of nuclei as well as intra-nuclear objects and their position within the nucleus. A step-by-step documentation is available for self-training, together with data sets of nuclei with different nuclear organization. Availability and implementation: Dataset of nuclei is available at https://www.gred-clermont.fr/media/WorkDirectory.zip. NucleusJ is available at http://imagejdocu.tudor.lu/doku.php?id=plugin:stacks:nuclear_analysis_plugin:start. Contact:philippe.andrey@versailles.inra.fr and christophe.tatout@univ-bpclermont.fr Supplementary information: Supplementary data are available at Bioinformatics online.

Plant tissue architecture and organ morphogenesis rely on the proper orientation of cell divisions. Previous attempts to predict division planes from cell geometry in plants mostly focused on 2D symmetric divisions. Using the stereotyped division patterns of Arabidopsis thaliana early embryogenesis, we investigated geometrical principles underlying plane selection in symmetric and in asymmetric divisions within complex 3D cell shapes. Introducing a 3D computational model of cell division, we show that area minimization constrained on passing through the cell centroid predicts observed divisions. Our results suggest that the positioning of division planes ensues from cell geometry and gives rise to spatially organized cell types with stereotyped shapes, thus underlining the role of self-organization in the developing architecture of the embryo. Our data further suggested the rule could be interpreted as surface minimization constrained by the nucleus position, which was validated using live imaging of cell divisions in the stomatal cell lineage. The proper positioning of division planes is key for correct development and morphogenesis of organs, in particular in plants were cellular walls prevent cell rearrangements. Elucidating how division planes are selected is therefore essential to decipher the cellular bases of plant morphogenesis. Previous attempts to identify geometrical rules relating cell shape and division plane positioning in plants mostly focused on symmetric divisions in tissues reduced to 2D geometries. Here, we combined 3D quantitative image analysis and a new 3D cell division model to evaluate the existence of geometrical rules in asymmetrical and symmetrical divisions of complex cell shapes. We show that in the early embryo of the model plant Arabidopsis thaliana, which presents stereotyped but complex cell division patterns, a single geometrical rule (area minimization constrained on passing through the cell centroid) recapitulates the complete sequence of division events. This new rule, valid for both symmetrical and asymmetrical divisions, generalizes previously proposed cell division rules and can be interpreted based on the dynamics of the cytoskeleton and on the positioning of the nucleus, a hypothesis that we validated using leaf cell division patterns. This work highlights the importance of self-organization in plant early morphogenesis and the emergence of robust cellular organizations based on geometrical feedback loops between cell geometry and division plane selection.

Studying how individual cells spatially and temporally organize within the embryo is a fundamental issue in modern developmental biology to better understand the first stages of embryogenesis. In order to perform high-throughput analyses in three-dimensional microscopic images, it is essential to be able to automatically segment, classify and track cell nuclei. Many 3D/4D segmentation and tracking algorithms have been reported in the literature. Most of them are specific to particular models or acquisition systems and often require the fine tuning of parameters.

ChIP-seq analyses of RBP LIF2 and its LHP1 partner in various backgrounds and stress conditions revealed target regions for the two proteins enriched in antagon