Yield losses from ozone pollution can be estimated by two methods: one involves the use of sensitive (S) and resistant (R) biotypes of white clover (Trifolium repens L., cv. Regal) exposed in ambient air, the other is based on the use of open-top chambers (OTC) supplied either with charcoal-filtered (CF) or non-filtered (NF) air. In southern Italy the two methods have been compared using the clover biotypes. The aim was (1) to compare the extent of ozone-induced yield reductions estimated by the two methods, (2) to evaluate the effect of the chamber enclosure on the growth of both biotypes, and (3) to compare plant water consumption in the different environments. On the average, the yield reduction was 23% when derived from the S/R yield ratio in ambient air, and 18% obtained by the CF/NF yield ratio of the S-type, without a significant difference between the two values. The slightly lower value for the OTC-based system may be due to the lower ozone levels in NF chambers due to losses in the ventilation system. Thus, both methods yielded equivalent yield reductions of about 20% due to ozone at this Mediterranean site. However, the higher air temperature inside OTCs influenced the plant growth, and this effect was stronger in the case of the R type. Therefore, R/S yield ratios in NF chambers differed from ambient air. Also, plant water consumption was higher in OTCs than in ambient air. The results suggest that the OTC-based method enables yield loss estimates at this Mediterranean site, in spite of chamber effects on plant growth and water use.