Abstract. The grain-producing regions of northern New South Wales and southern and central Queensland are characterised by cropping systems that are strongly dependent on stored soil moisture rather than in-crop rainfall, and tillage systems that are increasingly reliant on zero or minimum tillage. Crops are grown relatively infrequently and crop rotations are dominated by winter and summer grains (wheat [Triticum aestivum L.] and sorghum [Sorghum bicolor L. Moench], respectively), with smaller areas of grain legumes and cotton (Gossypium hirsutum L.). The grey, black, and brown Vertosols represent the more productive soils in the region under rainfed cropping, and are the focus of work reported in this study.Soil samples were collected from surface soils (0-0.30 m) across the region, utilising sites of long term tillage and residue management studies, fertiliser trials, and commercial fields to enable an assessment of the impact of various management practices on soil biological properties. A number of biological and biochemical parameters were measured (microbial biomass C, total organic C and labile C fractions, total C and N, microbial activity using FDA, cellulase activity, free living nematodes, total DNA and fatty acid profiles), and the response of wheat, sorghum, and chickpea (Cicer arietinum L.) to steam pasteurisation was assessed in glasshouse bioassays. The objective was to obtain an indication of the biological status of grain-growing soils and assess the impact of biological constraints in soils from different regions and management systems.Results showed that biological activity in cropped soils was consistently low relative to other land uses in northern Australia, with management practices like stubble retention and adoption of zero tillage producing relatively small benefits. In the case of zero tillage, many of these benefits were confined to the top 0.05 m of the soil profile. Fallowing to recharge soil moisture reserves significantly reduced all soil biological parameters, while pasture leys produced consistent positive benefits. Breaking a long fallow with a short duration grain or brown manure crop significantly moderated the negative effects of a long bare fallow on soil biology. Use of inorganic N and P fertilisers produced minimal effects on soil biota, with the exception of one component of the free-living nematode community (the Dorylaimida).The glasshouse bioassays provided consistent evidence that soil biota were constraining growth of both grain crops (sorghum and wheat) but not the grain legume (chickpea). The biota associated with this constraint have not yet been identified, but effects were consistent across the region and were not associated with the presence of any known pathogen or correlated with any of the measured soil biological or biochemical properties. Further work to confirm the existence and significance of these constraints under field conditions is needed.None of the measured biological or biochemical parameters consistently changed in response to management prac...