The effects of crop rotation and various cultural practices on soil organic matter and some biochemical characteristics of a heavy-textured, Orthic Black Chernozem with a thick A horizon were determined after 31 yr at Melfort, Saskatchewan. Treatments investigated included: fertilization, cropping frequency, green manuring, and inclusion of grass-legume hay crops in predominantly spring wheat (Triticum aestivum L.) systems. The results showed that neither soil organic C nor N in the top 15 cm of soil, nor hydrolyzable amino acids, nor C mineralized in 14 d at 20 °C were influenced by fertilization. However, the relative molar distribution (RMD) of the amino acids reflected the influence of fertilization and the phase (Rot-yr) of the legume green manure rotation sampled. Some characteristics assessed increased marginally with increasing cropping frequency but differences were less marked than results obtained earlier in a heavy-textured Black Chernozem with a thin A horizon at Indian Head, Saskatchewan. The relationship between soil organic matter or C mineralization versus estimated crop residues, residue C, or residue N returned to the land over the 31-yr period, were not significant in the Melfort soil. This contrasts with our findings for the thin Black soil. We speculate that the lack of soil organic matter response in the Melfort soil was due to its very high organic matter content (about 64 t ha−1C and 6.5 t ha−1N in the top 15 cm). We also hypothesized that the amino acid RMD results, which differed from most of those reported in the literature, may be reflecting the more recent cropping history of the soil. This aspect requires further research into the composition and distribution of the humic materials in this soil. Key words: Amino acids, relative molar distribution, C respiration, green manures, fertilization