Organic matter plays essential roles in soil, including physical stabilization, nutrient storage and carbon sequestration. An operationally defined fraction of soil organic matter known as glomalin or glomalin-related soil protein, GRSP, is obtained by autoclaving soil in citrate solution. It is reputed to be of fungal origin, very stable and responsible for enhanced soil physical stability. This is the first nationwide survey of GRSP content in soil and was conducted to test the hypotheses on the origin and identity of GRSP. Nearly 200 archived soils were selected on the basis of organic matter content from the French National Soil Inventory, representing mostly cropland, grassland and woodland land uses. Two fractions of GRSP were measured, easily extractable (GRSPEE) and total (GRSPT), extraction. The median values of GRSPEE and GRSPT were 0.7 and 2 g kg−1 respectively, with a strong correlation between the two fractions. Scatter plots and cubist modelling were used to explore the relationships between the contents of the two GRSP fractions and both soil properties and land use. Land use effects were almost entirely attributable to soil characteristics. No evidence was found to support the hypotheses that GRSP is solely of fungal origin, nor that easily extracted GRSP is more recent that the total fraction, although this does not disprove either hypothesis. Soil organic matter was enriched in GRSP C-depleted cropland soils and lower in C-rich woodland, this may result from inherent stability or differences in plant-related composition of carbon input.