Unknown,Transcriptomics,Genomics,Proteomics

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RNA-seq of sexually reproducing Pseudo-nitzschia multistriata diatom cells

ABSTRACT: Biological relevance and intent of the experiment: Given their abundance in the oceans and the exceptional adaptive capacities that make them capable of colonizing the most disparate habitats, diatoms constitute a group of microalgae of considerable ecological importance. This experiment aims to provide a molecular characterization of sexual reproduction for the diatom Pseudo-nitzschia multistriata, in relation to the consumption of nutrients and the impact on the marine ecosystem, by exploring the gene expression changes occurring at different time points in the cells involved in the process. Experimental workflow: Cell cultures of P. multistriata belonging to opposite mating types (MT- and MT+) were grown in 12L/12D cycles at 18°C under normal light conditions (60 uE) and crossed at a final cell density of 20.000 cells/ml using half volume from each MT. Samples for transcriptomic analyses were collected at three time points for sexually reproducing cells (1 hour: T1, 24 hours: T2 and 120 hours: T3), and at one time point (24 hours: T2) for parental monocultures grown in parallel as control condition.

INSTRUMENT(S): Illumina HiSeq 2000

ORGANISM(S): Pseudo-nitzschia multistriata

SUBMITTER: Mariella Ferrante

PROVIDER: E-MTAB-8854 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Pseudo-nitzschia multistriata is a planktonic diatom species with a diplontic life cycle comprising a short sexual phase during which gametes are produced from the encounter of two diploid cells of opposite mating type (MT). Gene expression studies have highlighted the presence of substantial changes occurring at the onset of sexual reproduction. Collectively, there is clear evidence of a chemical cross talk which mediates the mating step.Herein, we have hypothesized that these major transcriptomic changes are associated to variations in the amount and nature of the metabolites produced by the cells. In order to investigate such chemical diversity in an unbiased manner, we undertook an untargeted metabolomics approach. Liquid chromatography – tandem mass spectrometry was applied to investigate the differences in the metabolic profiles between control cells in the vegetative phase (MT+ and MT-) and mixed strains of opposite MTs (cross) undergoing sexual reproduction. Of the 2408 high-quality features obtained, 70 known metabolites could be identified based on in-house libraries and online databases; molecular networking proved valuable to classify additional 47 non-identified features.The present exploratory study provides an overview of the chemical diversity detectable in P. multistriata, presenting a dynamic picture of the changes occurring during sexual reproduction. In particular, the reduction of phytol detected in the cross can be linked to the general downregulation of photosynthesis during sexual reproduction observed elsewhere. The dysregulation of other metabolites stands as a starting point for the understanding of the mechanisms regulating sexual reproduction: a metabolite that was tentatively identified as 7-dehydrodesmosterol exhibited the highest upregulation in the comparison between the two MTs of opposite sex, as well as between MT+ vs cross.The effects of extraction solvents and harvesting times on the ensemble of endo-metabolites were also analyzed.

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RNA-seq of sexually reproducing Pseudo-nitzschia multistriata diatom cells

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