Description: It is well known that modern resting cysts with morphologies matching those of species of the fossil genus Spiniferites germinate into motile cells of the genus Gonyaulax. Different Spiniferites species have been connected to a single Gonyaulax species, raising the question of whether they are over-classified. Through germination experiments of cysts with the morphological features of four species of Spiniferites, viz. S. bentorii, S. hyperacanthus, S. ramosus and S. scabratus, we established cyst-theca relationships. Cysts with the morphology of S. bentorii gave rise to vegetative, motile cells of Gonyaulax nezaniae sp. nov., which is characterized by two stout antapical spines. Cysts with S. hyperacanthus and S. ramosus morphologies germinated into Gonyaulax whaseongensis and G. spinifera, respectively. Cysts with S. scabratus morphology lacked a ventral pore and were attributed to Gonyaulax cf. spinifera. Gene sequences for SSU, LSU and/or ITS-5.8S rRNA were obtained from these four species, and from cysts with the morphology of Spiniferites belerius, S. mirabilis, S. lazus, Spiniferites cf. bentorii and Tectatodinium pellitum. The maximum likelihood and Bayesian inference analyses based on LSU and SSU rRNA gene sequences revealed that cysts assignable to Spiniferites formed a polyphyletic group, intermingled with Tectatodinium, Bitectatodinium, Ataxiodinium and Impagidinium, whereas Gonyaulax species appeared as monophyletic. From our results we inferred the phylogenetic positions of S. bentorii, S. mirabilis, S. lazus, S. scabratus, Tectatodinium pellitum and Gonyaulax digitale for the first time, supporting the idea that Spiniferites species are not over-classified and each of them may correspond to different Gonyaulax species.

Description: The response of marine ecosystems to rapid climate changes has been well recognized but not studied extensively. Benthic microalgae, in contrast to the phytoplankton that is able to be transported by currents, have limited dispersal ability and thus are a better ecological indicator to climate changes. Here we performed sampling in the Yellow Sea, the East China Sea and South China Sea and established twenty-six strains of benthic Prorocentrum for detailed morphological and molecular examinations. Five Prorocentrum species, including P. concavum, P. fukuyoi, P. mexicanum, P. tsawwassenense, and P. cf. sculptile, were identified. Both P. concavum and P. fukuyoi displayed marked intraspecific divergences in large subunit (LSU) ribosomal RNA gene sequences, corresponding to their geographical origins. In contrast, P. mexicanum strains shared identical LSU sequence. Prorocentrum tsawwassenense and P. cf. sculptile are not suitable ecological indicators as they were rarely observed. Prorocentrum mexicanum is not recommended either as it is present across the region. In contrast, P. concavum and P. fukuyoi have advantages as ecological indicators for climate changes in the Western Pacific as they comprise several ribotypes with differentiated biogeography. Toxin analysis was also performed on all five species except P. fukuyoi by liquid chromatography coupled to tandem mass spectrometry, but okadaic acid was not detectable.