SNARE (soluble N‐ethylmaleimide‐sensitive factor attachment protein receptor) proteins are the principal actors of the concluding step of membrane traffic, the fusion of a vesicle with its acceptor membrane. The specificity of their interactions has been studied mostly in vitro and is now waiting for innovative approaches to elucidate the highly complex in vivo situations with which the multicellular organisms have satisfied the need for polarized and regulated secretion or vacuolar/lysosomal/endosomal trafficking. Plants, because of their high number of SNARE genes, their complex vacuolar system and peculiarity of established models for studying polarized and regulated exocytosis (pollen tube, root hair), represent a particularly challenging system for understanding the fine details of SNARE functions. The specificity of SNARE interactions resides in their structure but is certainly modulated by the cellular environment and a number of accessory factors; consequently, only in vivo experiments will reveal the full complexity of SNARE‐regulated processes. This review will highlight the major areas of interest and the approaches to study plant SNAREs specificity in vivo.