Regulation of vacuole fusion in stomata by dephosphorylation of the HOPS subunit VPS39

Understanding how plants regulate water loss is important for improving crop productivity. Tight control of stomatal opening and closing is essential for the uptake of CO2 while mitigating water vapor loss. The opening of stomata is regulated in part by homotypic vacuole fusion, which is mediated by conserved homotypic vacuole protein sorting (HOPS) and vacuolar SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptors) complexes. HOPS tethers apposing vacuole membranes and promotes the formation of trans-SNARE complexes to mediate fusion. In yeast, HOPS dissociates from the assembled SNARE complex to complete vacuole fusion, but little is known about this process in plants. HOPS-specific subunits VACUOLE PROTEIN SORTING39 (VPS39) and VPS41 are required for homotypic plant vacuole fusion, and a computational model predicted that post-translational modifications of HOPS may be needed for plant stomatal vacuole fusion. Here, we characterized a viable T-DNA insertion al..., , # Data from: Regulation of vacuole fusion in stomata by dephosphorylation of the HOPS subunit VPS39

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The methods for this dataset are described in detail in our manuscript. These compressed files contain:

Raw images (.czi) for vacuoles from roots (Root_vacuole_data.zip) used for Figure 1C.

Raw images (.czi) for stomata vacuoles (Stomata_Vacuole_Data.zip) used for Figure 1D-E and Figure 3D-E.

Images (.jpg) of siliques used for quantification of Figure 3A-C (Siliques_Data.tar). Genotypes associated with each plant number on each slide are listed in an Excel file.

qRT-PCR data (.xlsx) from seedlings corresponding to Figure 1B (Seedling_qRT_PCR_vps39-2.xlsx).

qRT-PCR data (.xlsx) from guard cell-enriched tissue corresponding to Figure 1F (Guard_Cell_enriched_RT_qPCR.xlsx).

## Description of the data and file structure

### **Root Vacuole image data files**

This includes confocal raw image files captured with a Zeiss LSM980 with Airy scan microscope. Images are organized in folde...,