In order to discriminate between detrimental, commensal, and beneficial microbes, plants rely on polysaccharides such as β-glucans, which are integral components of microbial and plant cell walls. The conversion of cell wall-associated β-glucan polymers into a specific outcome that affects plant-microbe interactions is mediated by hydrolytic and non-hydrolytic β-glucan-binding proteins. These proteins play crucial roles during microbial colonization: they influence the composition and resilience of host and microbial cell walls, regulate the homeostasis of apoplastic concentrations of β-glucan oligomers, and mediate β-glucan perception and signaling. This review outlines the dual roles of β-glucans and their binding proteins in plant immunity and symbiosis, highlighting recent discoveries on the role of β-glucan-binding proteins as modulators of immunity and as symbiosis receptors involved in the fine-tuning of microbial accommodation.