Drought-dependent regulation of cell coupling in Arabidopsis leaf epidermis requires plasmodesmal protein NHL12

The cytoplasm of most plant cells is connected by membrane-lined cell wall channels, the plasmodesmata (PD). Dynamic regulation of sugar, hormone, and protein diffusion through PD is essential for plant development and stress responses. Understanding this regulation requires knowledge of factors and mechanisms that control PD permeability through the modulation of callose levels in the cell wall around PD openings. We investigated PD regulation in leaf epidermal cells in relation to drought stress in Arabidopsis. PD-mediated cell wall permeability was decreased by drought stress and the hormone abscisic acid (ABA), and we tested how this related to several PD-associated genes with drought-responsive expression. Mutants of NON-RACE SPECIFIC DISEASE RESISTANCE/HIN1 HAIRPIN-INDUCED-LIKE 12 (NHL12) showed relatively low PD permeability that was unaffected by drought or ABA treatment. Overexpression of NHL12 in Nicotiana benthamiana epidermal cells increased PD permeability. Moreover, we showed that NHL12 can potentially interact with the callose synthase regulator NHL3 and we explored the effect of NHL12 abundance and/or lower interface permeability on ABA signaling genes. Our results indicate that NHL12 is a drought-responsive negative regulator of PD callose levels and, thereby, interface permeability. Results are discussed in relation to PD function during drought stress and the regulation of intercellular transport.