Fluorescence-based maximal quantum yield for PSII as a diagnostic of nutrient stress

In biological oceanography, it has been widely accepted that the maximum quantum yield of photosynthesis is influenced by nutrient stress. A closely related parameter, the maximum quantum yield for stable charge separation of PSII, (phi (PSII))(m), can be estimated by measuring the increase in fluorescence yield from dark-adapted minimal fluorescence (F-o) to maximal fluorescence (F-m) associated with the closing of photosynthetic reaction centers with saturating light or with a photosynthetic inhibitor such as 3 '-(3,4-dichlorophenyl)-1 ' ,1 ' -dimethyl urea (DCMU). The ratio F-v/F-m (= (F-m - F-o)/F-m) is thus used as a diagnostic of nutrient stress. Published results indicate that F-v/F-m is depressed for nutrient-stressed phytoplankton, both during nutrient starvation (unbalanced growth) and acclimated nutrient limitation (steady-state or balanced growth). In contrast to published results, fluorescence measurements from our laboratory indicate that F-v/F-m is high and insensitive to nutrient limitation for cultures in steady state under a wide range of relative growth rates and irradiance levels. This discrepancy between results could be attributed to differences in measurement systems or to differences in growth conditions. To resolve the uncertainty about F-v/F-m as a diagnostic of nutrient stress, we grew the neritic diatom Thalassiosira pseudonana (Hustedt) Hasle et Heimdal under nutrient-replete and nutrient-stressed conditions, using replicate semicontinuous, batch, and continuous cultures. Fv/Fm was determined using a conventional fluorometer and DCMU and with a pulse amplitude modulated (PAM) fluorometer. Reduction of excitation irradiance in the conventional fluorometer eliminated overestimation of F-o in the DCMU methodology for cultures grown at lower light levels, and for a large range of growth conditions there was a strong correlation between the measurements of F-v/F-m with DCMU and PAM (r(2) = 0.77, n = 460). Consistent with the literature, nutrient-replete cultures showed consistently high F-v/F-m (similar to0.65), independent of growth irradiance. Under nutrient-starved (batch culture and perturbed steady state) conditions, F-v/F-m was significantly correlated to time without the limiting nutrient and to nutrient-limited growth rate before starvation. In contrast to published results, our continuous culture experiments showed that F-v/F-m was not a good measure of nutrient limitation under balanced growth conditions and remained constant (similar to0.65) and independent of nutrient-limited growth rate under different irradiance levels. Because variable fluorescence can only be used as a diagnostic for nutrient-starved unbalanced growth conditions, a robust measure of nutrient stressed oceanic waters is still required.