title: INTERACTIVE EFFECTS OF SALINITY AND TEMPERATURE ON PLANOZYGOTE AND CYST FORMATION OF ALEXANDRIUM MINUTUM (DINOPHYCEAE) IN CULTURE(1).
abstract: The factors regulating dinoflagellate life-cycle transitions are poorly understood. However, their identification is essential to unravel the causes promoting the outbreaks of harmful algal blooms (HABs) because these blooms are often associated with the formation and germination of sexual cysts. Nevertheless, there is a lack of knowledge on the factors regulating planozygote-cyst transitions in dinoflagellates due to the difficulties of differentiating planozygotes from vegetative stages. In the present study, two different approaches were used to clarify the relevance of environmental factors on planozygote and cyst formation of the toxic dinoflagellate Alexandrium minutum Halim. First, the effects of changes in initial phosphate (P) and nitrate (N) concentrations in the medium on the percentage of planozygotes formed were examined using flow cytometry. Second, two factorial designs were used to determine how salinity (S), temperature (T), and the density of the initial cell inoculum (I) affect planozygote and resting-cyst formation. These experiments led to the following conclusions: 1. Low P/N ratios seem to induce gamete expression because the percentage of planozygotes recorded in the absence of added phosphate (-P) was significantly higher than that obtained in the absence of added nitrogen (-N), or when the concentrations of both nitrogen and phosphate were 20 times lower (N/20 + P/20). 2. Salinity (S) and temperature (T) strongly affected both planozygote and cyst formation, as sexuality in the population increased significantly as salinity decreased and temperatures increased. S, T combinations that resulted in no significant cyst formation were, however, favorable for vegetative growth, ruling out the possibility of negative effects on cell physiology. 3. The initial cell density is thought to be important for sexual cyst formation by determining the chances of gamete contact. However, the inoculum concentrations tested did not explain either planozygote formation or the appearance of resting cysts.
authors.affiliation: Institut de Ciénces del Mar (ICM-CSIC), Passeig Maritim de la Barceloneta 37-49, 08003 Barcelona, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, Spain.
authors.forename: Rosa Isabel
authors.initials: RI
authors.lastname: Figueroa
authors.affiliation: Institut de Ciénces del Mar (ICM-CSIC), Passeig Maritim de la Barceloneta 37-49, 08003 Barcelona, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, Spain.
authors.forename: Jose Antonio
authors.initials: JA
authors.lastname: Vázquez
authors.affiliation: Institut de Ciénces del Mar (ICM-CSIC), Passeig Maritim de la Barceloneta 37-49, 08003 Barcelona, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Massanet
authors.affiliation: Institut de Ciénces del Mar (ICM-CSIC), Passeig Maritim de la Barceloneta 37-49, 08003 Barcelona, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, Spain.
authors.forename: Miguel Anxo
authors.initials: MA
authors.lastname: Murado
authors.affiliation: Institut de Ciénces del Mar (ICM-CSIC), Passeig Maritim de la Barceloneta 37-49, 08003 Barcelona, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, SpainGrupo de Reciclado y Valorización de Materiales Residuales, Instituto de Investigaciones Marinas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, SpainInstituto Español de Oceanografía, Cabo Estai-Canido, 36200 Vigo, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Bravo
keywords: Alexandrium minutum; encystment; planozygotes; response surface methodology; salinity; sexuality; temperature