Different forms of plant biomass have been utilised for various applications in daily life and have gained increasing attention as replacements for fossil fuel-based products in the pursuit of a sustainable society. Plant cell walls, the primary carbon sink of plant biomass, have a high-order polysaccharide architecture consisting of cellulose, hemicelluloses, pectins, lignin and some proteins. Hemicelluloses are a group of polysaccharides that interact with cellulose, which is fundamental to the different properties and functionality of the plant cell walls. However, for industrial applications, the complex polysaccharide architecture poses a barrier to their efficient use. Understanding the molecular basis of plant cell walls - especially cellulose-hemicellulose interactions - is therefore critical to improving the utilisation of plant biomass. Recent research has revealed that the detailed structures, modification patterns, and conformation of hemicelluloses play an influential role in their interaction with cellulose. In this review, we discuss the latest insights into hemicelluloses across different forms of plant biomass and how their structures affect cell wall assembly. Additionally, we explore recent findings on how alterations in hemicellulose structure and modification patterns affect the usability of plant biomass, including the extractability of polysaccharides and the digestibility of biomass by glycoside hydrolases for biofuel production. Furthermore, we address unsolved questions in the field and propose future strategies to maximize the potential of plant biomass.