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Full length transcriptome sequencing of salivary glands of Pteromalus puparum

ABSTRACT: Animal saliva provides an excellent model for studying adaptive evolution, yet the functional significance of alternative mRNA isoforms in parasitoid salivary systems remains poorly understood. Here, using integrative full-length transcript sequencing and expression profiling, we generated an isoform-resolved transcriptomic landscape of salivary genes in the endoparasitoid wasp Pteromalus puparum. We identified 133 high-confidence salivary genes, more than 75% of which produced multiple transcript isoforms. Proteomic analyses confirmed that alternative splicing directly contributes to salivary protein diversity, with eight genes encoding distinct protein isoforms. Twelve salivary genes exhibited gland-biased isoform usage, including PpSerpin3, whose short isoform was specifically expressed in salivary and venom glands and suppressed host melanization. Comparative multi-omics analyses further revealed that salivary and venom systems share a conserved genetic core but achieve functional specialization through tissue-specific gene family co-option and extensive isoform switching. Together, these findings establish an isoform-resolved framework for the parasitoid salivary system and demonstrate that alternative splicing promotes salivary protein diversification and gland-specific functional evolution.

ORGANISM(S): Pteromalus puparum

PROVIDER: GSE330527 | GEO | 2026/05/13

REPOSITORIES: GEO

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Project description:Background The generalist dipteran pupal parasitoid Nasonia vitripennis injects 79 venom peptides into the host before egg laying. This venom induces several important changes in the host, including developmental arrest, immunosuppression, and alterations to normal metabolism. It is hoped that diverse and potent bioactivities of N. vitripennis venom provide an opportunity for the design of novel acting drugs. However, currently very little is known about the individual functions of N. vitripennis venom peptides and less than half can be bioinformatically annotated. The paucity of annotation information complicates the design of studies that seek to better understand the potential mechanisms underlying the envenomation response. Although the RNA interference system of N. vitripennis provides an opportunity to functionally characterise venom encoding genes, with 79 candidates this represents a daunting task. For this reason we were interested in determining the expression levels of venom encoding genes in the venom gland, such that this information could be used to rank candidate venoms. To do this we carried out deep sequencing of the transcriptome of the venom gland and neighbouring ovary tissue and used RNA-seq to measure expression from the 79 venom encoding genes. The generation of a specific venom gland transcriptome dataset also provides further opportunities to investigate novel features of this highly specialised organ. Results High throughput sequencing and RNA-seq revealed that the highest expressed venom encoding gene in the venom gland was a serine protease called Nasvi2EG007167, which has previously been implicated in the apoptotic activity of N. vitripennis venom. As expected the RNA-seq confirmed that the N. vitripennis venom encoding genes are almost exclusively expressed in the venom gland relative to the neighbouring ovary tissue. Novel peptides appear to perform key roles in N. vitripennis venom function as only four of the highest 15 expressed venom encoding genes are bioinformatically annotationed. The high throughput sequencing data also provided evidence for the existence of an additional 471 novel genes in the Nasonia genome that are expressed in the venom gland and ovary. Finally, metagenomic analysis of venom gland transcripts identified viral transcripts that may play an important part in the N. vitripennis venom function. Conclusions The expression level information provided here for the 79 venom encoding genes provides an unbiased dataset that can be used by the N. vitripennis community to identify high value candidates for further functional characterisation. These candidates represent bioactive peptides that have value in drug development pipelines.

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Full length transcriptome sequencing of salivary glands of Pteromalus puparum

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