Spatiotemporal Specific Blocking of Plasmodesmata by Callose Induction

Plasmodesmata are nanoscale cell wall channels connecting neighboring cells in plants. Intercellular trafficking of molecules via plasmodesmata plays important roles in various developmental processes and stress responses. The turnover of callose, a β-1,3-glucan polysaccharide depositing in the cell wall around plasmodesmata, controls the plasmodesmal permeability and symplasmic transport. Here, we describe a protocol for the spatiotemporally controlled induction of callose synthesis and plasmodesmata closure using the cals3m system. In this system, cals3m, a mutant CALLOSE SYNTHASE 3 (CALS3) gene, is driven by inducible tissue-specific promoters of interest. After appropriate induction by 17-β-estradiol, callose is overproduced within the corresponding specific domains, resulting in temporal closure of plasmodesmata at the cell-cell interfaces. This approach can be used to validate and dissect the function of plasmodesmata-mediated symplasmic communications.