Maximum photosynthesis rates in ferns are generally lower than those of seed plants, but little is known about the limiting factors, which are crucial to understand the evolution of photosynthesis in land plants. To address this issue, a gas exchange/chlorophyll fluorescence analysis was performed in three fern species spanning high phylogenetic range within Polypodiopsida (Osmunda regalis, Blechnum gibbum and Nephrolepis exaltata) to determine their maximum net photosynthesis (A(N)), stomatal (g(s)) and mesophyll (g(m)) conductances to CO2, and the maximum velocity of carboxylation (V-c,V-max). The in vitro Rubisco specificity factor (S-C/O) was also determined. All three species had values for S-C/O similar to those typical of seed plants, but values of A(N), g(s), g(m) and V-c,V-max were within the lowest range of those observed in seed plants. In addition, g(s) was unresponsive to light and CO2, as already described in other fern species. On the contrary, g(m) varied with changes CO2. A quantitative photosynthesis limitation analysis suggested that early land plants (ferns) presented not only stomatal limitationswhich were less adjustable to the environmentbut also restricted g(m) and V-c,V-max, resulting in limited maximum photosynthesis rates.