Pieter Gurdebeke

First record of resting cysts of the benthic dinoflagellate Prorocentrum leve in a natural reservoir in Gujan-Mestras, Gironde, France

The resting cysts of the benthic dinoflagellate Prorocentrum leve from a natural reservoir in Gujan‐Mestras (Gironde, France) were described in this study. The incubated urn‐shaped cysts gave rise to cells of P. leve. Morphological observations through light microscopy and scanning electron microscopy, particularly of the periflagellar platelets, combined with large subunit ribosomal DNA sequences obtained through single‐cell analysis confirm their affinity to the species P. leve. The cysts were characterized by a specific shape and the presence of an anterior plug. This is the first conclusive evidence for fossilizable resting stages within the Prorocentrales, one of the major orders within the Dinophyceae. Palynological treatments show that the cysts and endospores withstand hydrochloric and hydrofluoric acids. Micro‐Fourier transform infrared analysis on single specimens suggests that the composition of the endospore is cellulosic and the cyst wall a more robust, noncellulosic β‐glucan. The spectra overall are similar to other published spectra of resting cysts from autotrophic, planktonic dinoflagellates.

The affiliation of Hexasterias problematica and Halodinium verrucatum sp. nov. to ciliate cysts based on molecular phylogeny and cyst wall composition

Species in the genera Hexasterias and Halodinium have been recorded over the last decades as acritarchs in palynological and/or plankton studies. In paleoenvironmental studies, these resting stages are often interpreted as indicators of freshwater input. The biological affinity of these genera has never been definitely established. Here, a new species, Halodinium verrucatum sp. nov., is described and molecular evidence (single specimen SSU and LSU rDNA sequencing) reveals that both this new species and Hexasterias problematica, collected from sediment samples in the Skagerrak and Baltic Sea, are resting stages of prorodontid ciliates. Additionally, infrared spectroscopic analysis (micro-FTIR) of Hexasterias problematica and Halodinium spp. specimens indicates a carbohydrate-based composition of the cyst wall with evidence for nitrogen-containing functional groups. A similar composition is recorded for tintinnid loricae, which further supports the placement of Hexasterias and Halodinium as ciliate cysts, and the composition is consistent with the heterotrophic nature of ciliates. The morphologically similar species Radiosperma corbiferum has a comparable composition, suggesting a similar ciliate affinity and indicating the utility of micro-FTIR in understanding acritarch affinity. Hexasterias problematica typically occurs in coastal waters from temperate to arctic regions. Halodinium verrucatum sp. nov. is observed in temperate estuarine sediments in the northern hemisphere.

Morpho-molecular diversity and phylogeny of Bysmatrum (Dinophyceae) from the South China Sea and France

ABSTRACT The dinoflagellate genus Bysmatrum encompasses five epibenthic or tide-pool species and has been characterized by separated anterior intercalary plates. In the present study, we obtained six strains of Bysmatrum from the South China Sea and French Atlantic coast by isolating single cells/cysts from plankton and sediment samples. All strains were examined with light microscopy and scanning electron microscopy. Based on morphological observations, three strains were identified as Bysmatrum subsalsum, characterized by the elongated and rectangular first and a hexagonal second anterior intercalary plate. They differ from each other in the number of sulcal lists and the configuration of the first anterior intercalary plate. One strain was identified as Bysmatrum gregarium and the other two as Bysmatrum granulosum. The cyst-theca relationship of B. subsalsum from the French Atlantic was established by incubation of the cyst, and the geochemical composition of the cyst wall was measured through micro-Fourier transform infrared spectroscopy. Bysmatrum subsalsum from Malaysia shows a bright red stigma in the sulcal area under light microscopy, which was confirmed with transmission electron microscopy: it was identified as a type B eyespot. Small subunit ribosomal DNA (SSU rDNA), partial large subunit ribosomal DNA (LSU rDNA) and internal transcribed spacer (ITS) sequences were obtained from all six strains. The maximum likelihood and Bayesian inference analysis based on concatenated SSU, ITS and LSU sequences revealed that Bysmatrum is monophyletic and nested within Peridiniales. Our strains of B. subsalsum form a new ribotype in the molecular phylogeny (designated as ribotype B). The genetic distance based on ITS sequences among Bysmatrum species ranged from 0.34 to 0.47 and those genetic distances at the intraspecific level of B. subsalsum could reach 0.41, supporting the possibility of hidden crypticity within B. subsalsum.