Nature and effective range of non-cell autonomous activator and inhibitor peptides specifying plant stomatal patterning

Abstract

Stomata are epidermal valves that facilitate gas exchange between plants and their environment. Stomatal patterning is regulated by EPIDERMAL PATTERING FACTOR (EPF)-family of secreted peptides: EPF1 enforcing stomatal spacing, whereas EPF-LIKE9, also known as Stomagen, promoting stomatal development. It remains unknown, however, how far these signaling peptides act. Utilizing Cre-Lox recombination-based mosaic sectors that overexpress either EPF1 or Stomagen in Arabidopsis cotyledons, we reveal a range within the epidermis and across the cell layers in which these peptides influence patterns. To quantitatively determine their effective ranges, we developed a computational pipeline, SPACE (Stomata Patterning AutoCorrelation on Epidermis), that describes probabilistic two-dimensional stomatal distributions based upon spatial autocorrelation statistics used in Astrophysics. The SPACE analysis shows that, whereas both peptides act locally, the inhibitor, EPF1, exerts longer-range effects than the activator, Stomagen. Furthermore, local perturbation of stomatal development has little influence on global two-dimensional stomatal patterning. Our findings conclusively demonstrate the nature and extent of EPF peptides as non-cell autonomous local signals and provides a means to quantitatively characterize complex spatial patterns in development. Summary StatementNon-cell autonomous effects of activator and inhibitor peptides on 2-D spatial patterning of stomata were quantitatively characterized using chimeric sectors and a SPACE computational pipeline.