Abiotic stresses, such as drought, salinity, and extreme temperatures, have profound effects on plant reproduction, often leading to reduced fertility and yield. Reproduction in plants involves complex interactions between diverse cells, necessitating spatiotemporal resolution to understand how stress impacts each component of this intricate system. Imaging techniques have emerged as indispensable tools for uncovering the cellular and molecular responses of reproductive tissues to abiotic stresses in Arabidopsis and crops. Advanced methods, including fluorescence-based dyes and genetically encoded biosensors, have enabled the visualization of key stress-associated molecules such as reactive oxygen species and calcium ions. These approaches reveal the dynamic and localized nature of stress responses. Additionally, state-of-the-art imaging technologies, including light-sheet microscopy, structured illumination (e.g., Apotome), high-content confocal microscopy, micro-computed tomography, and custom heated-stage setups, provide varying levels of spatial and temporal resolution to study stress-induced changes in tissue morphology and development. Complementary techniques like sectioning and staining continue to yield critical insights into the anatomical and developmental alterations under stress conditions. This review integrates findings from these methodologies, highlighting their contributions to our understanding of abiotic stress responses in male and female reproductive tissues. Furthermore, we identify technological advancements needed to enable real-time, (sub)cellular-level imaging of stress responses. Finally, we compile a list of promoter-based identity markers specific to reproductive tissues across different crop species, offering a resource for targeted genetic studies. By bridging current imaging techniques with biological insights and technological gaps, this work aims to advance the field of plant stress biology and reproductive resilience.

Centro de Recursos Naturales Renovables de la Zona Semiárida CONICET Bahía Blanca Buenos Aires Argentina

Department of Biosciences University of Milan Milano Italy

Department of Biosystems KU Leuven Leuven Belgium

Laboratory of Functional Genomics and Proteomics National Centre for Biomolecular Research Faculty of Science Masaryk University Brno Czech Republic

Leuven Plant Institute KU Leuven Leuven Belgium

Mendel Center for Plant Genomics and Proteomics Central European Institute of Technology Masaryk University Brno Czech Republic

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