Morphological and LSU rDNA sequence variation within the Gonyaulax spinifera-Spiniferites group (Dinophyceae) and proposal of G. elongata comb. nov. and G. membranacea comb. nov.

Cultures were established from cysts of the cyst-based taxa Spiniferites elongatus and S. membranaceus. Motile cells and cysts from both cultures and sediment samples were examined using light and scanning electron microscopy. The cyst-theca relationship was established for S. elongatus. The motile cells have the tabulation pattern 2 pr, 4′, 6″, 6c, ≥ 4s, 6‴, 1p, 1 ‴, but they remain unattributable to previously described Gonyaulax species. There was large variation in process length and process morphology in cysts from both cultures and wild samples and there was variation in ornamentation and in the development of spines and flanges in motile cells. A new combination, G. elongata (Reid) Ellegaard et al. comb. nov. is proposed, following new rules of the International Code of Botanical Nomenclature that give genera based on extant forms priority over genera based on fossil forms. Extreme morphological variation in the cyst and motile stages of S. membranaceus is described and this species is also transferred to the genus Gonyaulax, as G. membranacea (Rossignol) Ellegaard et al. comb. nov. Approximately 1500 bp of large subunit (LSU) rDNA were determined for these two species and for G. baltica, G. cf. spinifera (= S. ramosus) and G. digitalis (= Bitectatodinium tepikiense). LSU rDNA showed sequence divergences similar to those estimated between species in other genera within the Gonyaulacales; a phylogeny for the Gonyaulacales was established, including novel LSU rDNA sequences for Alexandrium margalefii, A. pseudogonyaulax and Pyrodinium bahamense var. compressum. Our results show that motile stages obtained from the germination of several cysts of the 'fossil-based' Spiniferites and B. tepikiense, which were previously attributed to 'Gonyaulax spinifera group undifferentiated', belong to distinct species of the genus Gonyaulax. These species show small morphological differences in the motile stage but relatively high sequence divergence. Moreover, this group of species is monophyletic, supported by bootstrap values of 100% in parsimony and maximum likelihood analyses.