Grain filling is an intensive transportation process regulated by plant hormones. The present study investigated whether and how the interaction between abscisic acid (ABA) and ethylene is involved in mediating the grain filling of rice (Oryza sativa L.) under non-flooded mulching cultivation. A field experiment repeated over 2 years was conducted with two high-yielding rice cultivars, Zhendao 88 (a japonica cultivar) and Shanyou 63 (an indica hybrid cultivar), and four cultivation treatments were imposed from transplanting to maturity: traditional flooding as control (TF), non-flooded plastic film mulching (PM), non-flooded wheat straw mulching (SM) or non-flooded no mulching (NM). Compared with that under TF, grain yield was reduced by 21·0–23·1% under PM (P<0·05), 1·4–1·8% under SM (P>0·05) and 50·9–55·4% under NM (P<0·05). Both PM and NM significantly (P<0·05) reduced the proportion of filled grains and grain weight and were associated with decreased grain filling rates. In SM there was a significant increase in the grain filling rate. The concentration of ABA in the grains was very low at the early grain filling stage, reaching a maximum when the grain filling rate was the highest, and showed no significant differences (P>0·05) between TF, PM and SM. However, it was significantly higher in NM. In contrast to ABA, the ethylene evolution rate and 1-aminocyclopropane-1-carboxylic acid (ACC) concentration in the grains were very high at the start of grain filling and sharply decreased during the active grain filling period. Both PM and NM increased the ethylene evolution rate and ACC concentration, whereas these were reduced in SM. The ratio of ABA to ACC was increased under SM but decreased under PM and NM, indicating that ethylene was more enhanced than ABA when plants were grown under NM and PM. The concentration of ABA correlated with the grain filling rate as a hyperbolic curve, whereas the ethylene evolution rate correlated with the grain filling rate as an exponential decay equation. The ratio of ABA to ACC significantly correlated with the grain filling rate with a linear relationship. Application of amino-ethoxyvinylglycine (inhibitor of ethylene synthesis by inhibiting ACC synthase) or ABA to panicles under TF and PM at the early grain filling stage significantly increased activities of the key enzymes involved in sucrose to starch conversion in the grains, sucrose synthase, ADP glucose pyrophosphorylase and soluble starch synthase, grain filling rate and grain weight. Application of ethephon (ethylene-releasing agent) or fluridone (inhibitor of ABA synthesis) had the opposite effect. The results suggest that antagonistic interactions between ABA and ethylene may be involved in mediating the effect of non-flooded mulching cultivation on grain filling, and a high ratio of ABA to ethylene enhances grain filling rate.