FLAVODOXIN EXPRESSION AS AN INDICATOR OF IRON LIMITATION IN MARINE DIATOMS¹

We have prevously shown that a marine chlorophyte expressed flavodoxin under iron limitation but not under other nutrient stress conditions. Here we use polyclonal antiserum raised against the diatom Phaeodactylum tricornutum Bohlin to show that a similar response is observed in this species. Using our antibody, western blotting techniques, and standard colorimetric detection (4‐chloro‐1‐naphthol), we can detect at least a 25–50‐fold increase in flavodoxin in iron‐depleted compared to iron‐replete cells. In iron‐limited batch cultures of P. tricornulum, flavodoxin accumulation was inversely proportional to growth rate and was not detectable in cultures containing initially more than 750 nm of iron. We demonstrated that the accumulation of flavodoxin under iron stress is widespread among marine diatoms and that it may be possible to use the presence or absence of flavodoxin in natural marine diatom assemblages to detect iron limitation. However, our polyclonal antisera appears to be specific for diatoms and did not cross‐react with Synechococcus sp., Micromonas pusilla (Butcher) Manton et Parke, Dunaliella tertiolecta Butcher, Chlorella sp., Emiliania huxleii (Lohm.) Hay et Parke, or Isochrysis galbana Parke. A reverse bioassay experiment was conducted with natural phytoplankton assemblages containing mainly diatoms from Long Island Sound and in shelf waters near Cape Hatteras, two areas not suspected to be iron‐limited. Although flavodoxin was not detected in situ in these areas, natural populations of diatoms driven into iron limitation expres.sed flavodoxin. Flavodoxin was detected in mats of the diatom Rhizosolenia castracanei Cleve collected from the Equatorial Pacific during a JGOFS cruise in 1992, consistent with the hypothesis that iron may be limiting in this high‐nutrient, low‐chlorophyll region.