Aims: The aim of the present study was to compare lead accumulation and tolerance among heavy metal hyperaccumulating and non-hyperaccumulating metallophytes. Methods: To this purpose, we compared Pb tolerance and accumulation in hydroponics among calamine and non-calamine populations of Silene vulgaris, Noccaea caerulescens, and Matthiola flavida. We established the effects of Ca on Pb tolerance and accumulation in M. flavida, and measured exchangeable soil Pb and Ca at two calamine sites. Results: Results revealed that calamine populations of S. vulgaris and N. caerulescens were Pb hypertolerant, but the calamine M. flavida population was not. Pb hyperaccumulation capacity was exclusively found in one of the calamine N. caerulescens populations. Conclusions: 1) Pb hypertolerance is sometimes lacking in metallophyte populations from strongly Pb-enriched soil, probably due to a relatively high level of exchangeable soil Ca, 2) Ca effectively counteracts Pb uptake and Pb toxicity, 3) The tendency to hyperaccumulate Pb is a population-specific phenomenon in N. caerulescens, 4) Pb hypertolerance in N. caerulescens is not necessarily associated with a tendency to hyperaccumulate Pb, 5) apparent natural Pb hyperaccumulation in M. flavida is not reproducible in hydroponics, probably due to the absence of air-born contamination in laboratory experiments. © 2011 Springer Science+Business Media B.V.