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Five Non-motile Dinotom Dinoflagellates of the Genus Dinothrix.

ABSTRACT: Dinothrix paradoxa and Gymnodinium quadrilobatum are benthic dinoflagellates possessing diatom-derived tertiary plastids, so-called dinotoms. Due to the lack of available genetic information, their phylogenetic relationship remains unknown. In this study, sequencing of 18S ribosomal DNA (rDNA) and the rbcL gene from temporary cultures isolated from natural samples revealed that they are close relatives of another dinotom, Galeidinium rugatum. The morphologies of these three dinotoms differ significantly from each other; however, they share a distinctive life cycle, in which the non-motile cells without flagella are their dominant phase. Cell division occurs in this non-motile phase, while swimming cells only appear for several hours after being released from each daughter cell. Furthermore, we succeeded in isolating and establishing two novel dinotom strains, HG180 and HG204, which show a similar life cycle and are phylogenetically closely related to the aforementioned three species. The non-motile cells of strain HG180 are characterized by the possession of a hemispheroidal cell covered with numerous nodes, while those of the strain HG204 form aggregations consisting of spherical smooth-surface cells. Based on the similarity in life cycles and phylogenetic closeness, we conclude that all five species should belong to a single genus, Dinothrix, the oldest genus within this clade. We transferred Ga. rugatum and Gy. quadrilobatum to Dinothrix, and described strains HG180 and HG204 as Dinothrix phymatodea sp. nov. and Dinothrix pseudoparadoxa sp. nov.

SUBMITTER: Yamada N

PROVIDER: S-EPMC7710806 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The genus Lactobacillus contains over 100 different species that were traditionally considered to be uniformly non-motile. However, at least twelve motile species are known to exist in the L. salivarius clade of this genus. Of these, Lactobacillus rumnis is the only motile species that is also autochthonous to the mammalian gastrointestinal tract. The genomes of two L. ruminis strains, ATCC25644 (human isolate, non-motile) and ATCC27782 (bovine isolate, motile) were sequenced and annotated to identify the genes responsible for flagellum biogenesis and chemotaxis in this species. Transcriptome analysis revealed that motility genes were transcribed at a significantly higher level in motile L. ruminis ATCC27782 than in non-motile ATCC25644 during the motile growth phase. In preparation for RNA isolation from L. ruminis ATCC27782 and ATCC25644 cultures in the motile phase, 20 ml aliquots of MRS broth were inoculated (0.25 % inoculum) with a turbid culture of the desired strain. The cultures were incubated anaerobically at 37 Â°C for 15 hr. In preparation for RNA isolation from L. ruminis ATCC27782 and ATCC25644 cultures in the non-motile growth phase, 20 ml aliquots of MRS broth were inoculated (1 % inoculum) with a turbid culture of the desired strain. The cultures were incubated anaerobically at 37 Â°C for 16-20 hrs. RNAprotect Bacteria reagent (Qiagen) was used to stabilize gene expression in the target cultures, and total RNA was isolated according to the protocol for enzymatic and mechanical disruption of bacteria as described in the RNAprotect Bacteria Reagent handbook. RNA isolation was completed with the RNeasy Mini kit, and contaminating DNA was removed with the Turbo DNA-free kit (Ambion). An Oligo aCGH/ChIP-on chip hybridization kit (Agilent) was used for hybridization of the labelled cDNA to the microarrays. Probe hybridization took place at 65 Â°C for 20 hrs with constant rotation (10 rpm). Microarrays were scanned using the Agilent Microarray Scanner System (G2505B) and the scanned files were converted to data files with Feature Extraction software (Agilent). Three microarrays, (three biological replicates) were used to examine gene transcription during the motile growth phase. One microarray was used to examine gene transcription during the non-motile growth phase.

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Five Non-motile Dinotom Dinoflagellates of the Genus Dinothrix.

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