Stomata are microscopic valves present on plant aerial surfaces for gas exchange, which helps in facilitating photosynthesis and transpiration. Significant advances have been made in the molecular mechanism of stomatal development in the last two decades, which often provides an insight into plant survival and growth. Many new genes were identified by traditional forward genetic screening, microarray analysis between various mutants, or protein-protein interaction methods. The role of functional redundant family members was specified using protein domain swapping approaches or generating dominant-negative mutants. The target genes of transcription factors were identified by combining RNA-seq analysis of the inducible transcription factors and ChiP-seq assays. The spatiotemporal function of polarized proteins in transient cell states was studied using a long-term time-lapse imaging approach. The recognition of specific ligands by a receptor complex was identified by manipulating the purified ligands and the binding domains of the receptor complex using biophysical and biochemical approaches. This Methods Collection brings together the wide range of techniques that have been developed to study stomatal development. The goal of this Methods Collection is to provide detailed state-of-the-art protocols and procedures currently in use in the field of stomatal development.

Examples of methods included in stomatal development are microarray, protein domain swapping study, gain-of-function protein study, dominant-negative protein study, ChiP study, RNA-seq study, pull-down assays using fusion proteins, proximity-dependent labeling techniques (e.g., TurboID interactions), yeast two-hybrid technologies, protein complementation assays (e.g., BiFC), biophysical approaches (X-ray crystallography, NMR), and purification of small peptide ligands.