Genetic encoding of climate-responsive stomatal developmental plasticity in tomato

Flexible developmental programs enable plants to customize their organ size and cellular composition. In leaves of eudicots, the stomatal lineage produces two essential cell types, stomata and pavement cells, and plants can adjust the total numbers and ratios of these cell types in response to external cues. Central to this flexibility is the stomatal lineage-initiating transcription factor, SPEECHLESS (SPCH). Here we explore the mechanisms underlying SPCH’s involvement in environmental response. Using multiplexed CRISPR/Cas9 editing of SlSPCH cis- regulatory sequences in tomato, we identified variants with altered stomatal development responses to drought, light and temperature cues. By creating and live-cell tracking translational reporters of SlSPCH and its paralogues SlMUTE and SlFAMA, we revealed the corresponding cellular events that lead to the environmental change-driven responses in stomatal production and leaf form. Plants bearing the novel reporters and SlSPCH variants are powerful resources for fundamental and applied studies of tomato resilience in response to climate change.