Plant cell walls are complex configurations of polysaccharides that fulfil a diversity of roles during plant growth and development. They also form sets of biomaterials that are widely exploited in food, fibre and fuel applications. The pectic polysaccharides, that comprise approximately a third of primary cell walls, form complex supramolecular structures comprised of distinct glycan domains. Rhamnogalacturonan-I (RG-I) is a highly structurally heterogeneous branched glycan domain within the pectic supramolecule that contains rhamnogalacturonan, arabinan and galactan as structural elements. Heterogeneous RG-I polymers are implicated in generating the mechanical properties of cell walls during cell development and plant growth but are poorly understood in architectural, biochemical and functional terms. Using specific monoclonal antibodies to the three major RG-I structural elements (arabinan, galactan and the rhamnogalacturonan backbone) for in situ analyses, and also for chromatographic detection analyses, the relative occurrences of RG-I structures have been studied within a single tissue - the tobacco seed endosperm. Analyses indicate that RG-I polymer features display spatial heterogeneity at the level of the tissue, at the level of single cell walls and also heterogeneity at the biochemical level. This work has implications for understanding RG-I glycan complexity in the context of cell wall architectures and in relation to cell wall functions in cell and tissue development. This article is protected by copyright. All rights reserved.