Barry C. Smith

Dinoflagellate cyst production in one-liter containers

Methods for the production of dinoflagellate cysts in two types of 1 L containers have been developed. Using these methods, dinoflagellate cysts can be produced in amounts large enough for shellfish grazing experiments or whenever large amounts of cysts are needed. The species used were Scrippsiella lachrymosa (B-10) and toxic Alexandrium fundyense (CB501 and GTM25). Cultures of S. lachrymosa yielded 628 ± 74 cysts mL−1 and A. fundyense cultures yielded 350 ± 98 cysts mL−1. Findings suggest that aspects of the boundary layer between the media and the wall of the container are important for gamete mating; especially, the slope of the container wall appears to be relevant, which offers some explanation of previous observations that the shape of the container is important in the formation of dinoflagellate resting cysts. These observations may support the theory that physical interfaces in nature facilitate dinoflagellate encystment.

Synchronization of encystment of Scrippsiella lachrymosa (Dinophyta)

The encystment of Scrippsiella lachrymosa cells (strain B-10), which can be induced reliably in encystment medium, was inhibited by stirring the culture. 100 mL cultures in glass beakers were stirred at 1 rotation s−1. Stirring inhibited vegetative cells from congregating (swarming) at the walls of the culture container. When stirring was stopped, a rapid induction of sexual reproduction was seen. As soon as stirring stopped (within 2 min), cells were observed swarming near the edges of the glass beaker. Four days after cessation of stirring, large percentages of the cells were mating and, after 7 days, most were zygotes. Cultures were observed after 31, 38, and, 45 days of stirring. When cultures were stirred for 45 days, cysts developed in the stirred treatments, but these cysts were attached to flocculent material that had also formed in the medium. The use of this laboratory method is advantageous for the study of the mating through cyst stages of the dinoflagellate life history. This method may also demonstrate the need for a ‘surface’ as a place for the dinoflagellate to congregate in order to successfully encyst and may help explain environmental observations of encystment at pycnoclines.

Nutrient interactions between phytoplankton and bacterioplankton under different carbon dioxide regimes

Light, nutrients, temperature, pH, and salinity are important factors in controlling the growth of phytoplankton and bacterioplankton. Supply of key nutrients to these communities can result in mutualistic or competitive relationships between bacterioplankton and phytoplankton. In this study, we investigated growth and uptake of nutrients by the marine prasinophyte flagellate Tetraselmis chui (strain PLY429) in the presence and absence of a community of bacterioplankton at two pH levels. Growth of PLY429 and total nutrient uptake were calculated for each treatment. The addition of bacterioplankton resulted in lower growth rates of PLY429, but the removal of ammonium was greater in those cultures with bacterioplankton present. The division rate of PLY429 was affected by pH; however, pH changes did not result in different uptake rates of nitrate, ammonium, or phosphate by the mixed algal and bacterial assemblage. These findings suggest that bacterioplankton and phytoplankton were competing for ammonium and that a lower pH resulted in more rapid algal growth.

Differences in pigmentation between life cycle stages in Scrippsiella lachrymosa (dinophyceae).

Various life cycle stages of cyst‐producing dinoflagellates often appear differently colored under the microscope; gametes appear paler while zygotes are darker in comparison to vegetative cells. To compare physiological and photochemical competency, the pigment composition of discrete life cycle stages was determined for the common resting cyst‐producing dinoflagellate Scrippsiella lachrymosa. Vegetative cells had the highest cellular pigment content (25.2 ± 0.5 pg · cell−1), whereas gamete pigment content was 22% lower. The pigment content of zygotes was 82% lower than vegetative cells, even though they appeared darker under the microscope. Zygotes of S. lachrymosa contained significantly higher cellular concentrations of β‐carotene (0.65 ± 0.15 pg · cell−1) than all other life stages. Photoprotective pigments and the de‐epoxidation ratio of xanthophylls‐cycle pigments in S. lachrymosa were significantly elevated in zygotes and cysts compared to other stages. This suggests a role for accessory pigments in combating intracellular oxidative stress during sexual reproduction or encystment. Resting cysts contained some pigments even though chloroplasts were not visible, suggesting that the brightly colored accumulation body contained photosynthetic pigments. The differences in pigmentation between life stages have implications for interpretation of pigment data from field samples when sampled during dinoflagellate blooms.