Light response curve methodology and possible implications in the application of chlorophyll fluorescence to benthic diatoms

Chlorophyll a fluorescence has been increasingly applied to benthic microalgae, especially diatoms, for measurements of electron transport rate (ETR) and construction of rapid light response curves (RLCs) for the determination of photophysiological parameters [mainly the maximum relative ETR (rETR(max)), the light saturation coefficient (E (k)) and the maximum light use coefficient (alpha)]. Various problems with the estimation of ETR from the microphytobenthos have been identified, especially in situ. This study further examined the effects of light history of the cells and light dose accumulation during RLCs on the fluorescence measurements of ETR using the benthic diatom Navicula phyllepta. RLCs failed to saturate when using incremental increases in irradiance, however, curves with decreasing irradiance did saturate. Patterns indicating photoacclimation in response to light histories were observed, with higher rETR(max) and E (k), and lower alpha, at high light compared to low light. However, these differences could be negated by increasing the RLC irradiance duration from 30 to 60 s. It is suggested that problems arose as a result of rapid fluorescence variations due to ubiquinone (Q(A)) oxidation and non-photochemical chlorophyll fluorescence quenching (NPQ) which depended upon the light history of the cells and the RLCs accumulated light dose. Also, RLCs with irradiance duration of 10 s were shown to have a high level of error possibly specific to the fluorimeter programming. It is suggested that RLCs, using a Diving-PAM fluorimeter on benthic diatoms, should be run using decreasing irradiance steps of 30 s duration.