Unknown,Transcriptomics,Genomics,Proteomics

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Bacterial endosymbionts control host sexuality and reveal reproductive genes of early divergent fungi

ABSTRACT: Through transcriptome profiling using RNA-seq, we investigated the mechanisms behind bacterial endosymbiont (Burkholderia rhizoxinica) control over host (Rhizopus microsporus) reproductive biology. By analyzing differential expression across six different conditions, including fungal opposite mates growing independently with or without endosymbionts, as well as opposite mates growing together with endosymbionts (mating) or without endosymbionts (no mating), we were able to identify that endosymbionts control expression of a Ras signaling protein critical for sexual reproduction in many fungi (Ras2). As little is known regarding sexual reproduction in Mucoromycotina, we also used these data to investigate conservation of sex-related genes across all fungi, as well as predict potential genes involved in sensing of trisporic acid, the mating pheromone used by these fungi. 6 different conditions were analyzed, each consisting of two biological replicates. These included Rhizopus microsporus ATCC52813 (sex +) growing alone with endosymbionts, R. microsporus ATCC52814 (sex -) growing alone with endosymbionts, ATCC 52813 growing alone without endosymbionts, ATCC52814 growing alone without endosymbionts, ATCC52813 and ATCC52814 growing together with endosymbionts (successfully mating), and ATCC52813 and ATCC52814 growing together without endosymbionts (failure to mate). In each condition, fungi were cultivated on half-strength PDA and plugs of mycelium were placed at the edge of the plate. After 6 days, approximately 2.5 cm of tissue were harvested from the center of the plate. Each biological replicate consists of 5 plates which were pooled prior to RNA extraction to ensure sufficient tissue was collected.

ORGANISM(S): Rhizopus microsporus

SUBMITTER: Stephen Mondo

PROVIDER: E-GEOD-57644 | 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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Bacterial endosymbionts control host sexuality and reveal reproductive genes of early divergent fungi

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