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ABSTRACT: Microbiome repertoire across the life cycle of the milkweed bug Oncopeltus fasciatus (16S amplicon sequencing)

PROVIDER: PRJNA1213894 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR32073024_1.fastq.gz	Fastqsanger.gz
	SRR32073024_2.fastq.gz	Fastqsanger.gz
	SRR32073025_1.fastq.gz	Fastqsanger.gz
	SRR32073025_2.fastq.gz	Fastqsanger.gz
	SRR32073026_1.fastq.gz	Fastqsanger.gz

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Project description:Despite considerable speculation for the role of cytosine (DNA) methylation in biological and molecular processes in insects, direct functional tests are lacking. Here we provide evidence for the functional role of the maintenance DNA methyltransferase 1 (Dnmt1) in an insect using experimental manipulation. Through RNA interference (RNAi) we successfully post-transcriptionally knocked down Dnmt1 in ovarian tissue of the hemipteran Oncopeltus fasciatus (the large milkweed bug). Individuals depleted for dnmt1, and subsequently DNA methylation, failed to reproduce. Manipulating the levels of DNA methylation did not result in changes in overall gene expression. Furthermore, reductions in levels of DNA methylation at transposable elements (TEs) did not lead to large-scale reactivation of TE transcription. Despite the lack of a causal relationship between reduced DNA methylation and gene expression in the tissue we surveyed, eggs were inviable revealing an important function of DNA methylation in O. fasciatus. Our work provides direct experimental evidence for a functional role of Dnmt1 and DNA methylation in insects and presents O. fasciatus as a tractable model for further exploration of the function of DNA methylation in other tissues and life history circumstances for insects.

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Project description:The unparalleled success of the insects comprising more than a million species has long stood out to evolutionary biologists. A much overlooked evolutionary innovation of the insects is the serosa, an extraembryonic epithelium that covers yolk and embryo in their eggs. We have shown that this epithelium provides innate immune protection to eggs of the beetle Tribolium castaneum. It remained elusive, however, if this innate immune competence evolved in the Tribolium lineage, or is ancestral to all insects. Here, we expand our studies to the bug Oncopeltus fasciatus that belongs to the basal main group of insects, the Hemimetabola. RNA sequencing reveals an extensive transcriptional response upon infection of the egg with Gram-positive and Gram-negative bacteria. We demonstrate the antimicrobial activity of upregulated peptides using in vitro bacterial growth inhibition assays, and describe two novel families of AMPs called Serosins and Ovicins. By qPCR, we determine that eggs become immune responsive when the serosa develops. Finally, in situ hybridizations show that transcripts of upregulated peptides are located in the serosal cells and not in the underlying embryo. We conclude that the serosa protects the O. fasciatus embryo against pathogens. This first evidence from hemimetabolous insect eggs suggests that immune competence is an ancestral property of the serosa. The evolutionary origin of the serosa with its immune function might have been one of the factors that facilitated the spectacular success of the insects.

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