Some biological and chemical properties of a Gleysol were examined after 24 years of soil tillage (chisel plough – CP, mouldboard plough – MP, no-till – NT) and that of ploughing the 24-yr NT (P-NT) once, in two cropping systems (conventional – CONV, organic – ORG) applied over 4 years (2007–2010) of a long-term experiment (autumn 1987–autumn 2011) at La Pocatière, Québec, Canada. The 0–10, 10–20 and 20–30 cm soil depths were sampled in autumn 2011 after a maize trial. Tillage affected light fraction organic carbon (LFOC), light fraction organic nitrogen (LFON) and mineralizable N (Nmin) in soil, with the lowest LFOC, LFON and Nmin values in the MP treatment. No-till had lower soil pH than the other tillage systems in the 10–20 and 20–30 cm soil depths. Tillage affected the amounts of nitrate-N in 0–10 and 10–20 cm soil depths, with the lowest amounts for MP (4.3 kg nitrate-N/ha) compared with NT (7.2 or 8.5 kg nitrate-N/ha) or CP (7.7 kg nitrate-N/ha). The P-NT had no negative impact on organic C and N, or available nutrients in soil. Cropping system had no effect on soil organic C and N, available nutrients or pH. Findings suggest that long-term NT or CP may result in greater storage of organic C and N in soil and improve available nutrients compared with MP. Ploughing 25-year-old NT plots redistributed available nutrients in the profile but had no negative effect on soil organic C or N.