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Transcriptomic Analyses of Secreted Proteins From the Salivary Glands of Wheat Midge Larvae.

ABSTRACT: AbstractBoth the wheat midge () (Géhin) (Diptera: Cecidomyiidae) and the Hessian fly () (Say) (Diptera: Cecidomyiidae) belong to a group of insects called gall midges (Diptera: Cecidomyiidae), and both are destructive pests of wheat. From Hessian fly larvae, a large number of genes have been identified to encode secreted salivary gland proteins (SSGPs), which are presumably critical for the insect to feed on and manipulate host plants. For comparison, we conducted an analysis on transcripts encoding SSGPs from the first instar larvae of the wheat midge. In total, 3,500 cDNA clones were sequenced, from which 1,301 high-quality sequences were obtained. Approximately 25% of the cDNAs with high-quality sequences encoded SSGPs. The SSGPs were grouped into 97 groups based on sequence homology. Among the SSGP-encoding transcripts, 206 encoded unique proteins with no sequence similarity to any known protein and 29 encoded proteins similar to known proteins including proteases, serpines, thioesterases, ankyrins, and ferritins. Most (~80%) SSGP-encoding genes appear under strong selection for mutations that generate amino acid changes within the coding region. Identification and characterization of SSGPs in wheat midge larvae provide a foundation for future work to reveal molecular mechanisms behind wheat midge–wheat interactions and the role of these putative effector proteins in insect virulence. Availability of the SSGP transcripts will also facilitate comparative analyses of insect effectors from related species.

SUBMITTER: Al-jbory Z

PROVIDER: S-EPMC5822882 | biostudies-other | 2018 Jan

REPOSITORIES: biostudies-other

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Project description:Although the salivary transcriptome of adult mosquitoes has been thoroughly described in several recent papers, very little information exists regarding the biological role of larval salivary glands in the Culicidae. We used whole-transcriptome AffymetrixÂ® chips to compare the transcriptional profiles of Anopheles gambiae larval (L4) salivary glands and whole larvae. We identified a total of 277 transcripts as being significantly enriched in the salivary glands. Based on available annotation for the known or predicted protein sequences encoded by these transcripts, 41 were identified as corresponding to secreted proteins, 233 as corresponding to non-secreted (housekeeping) proteins, and 3 as encoding proteins of unknown function. Based on functional annotation of the putatively secreted gene products, we propose that larval salivary secretions have roles in nutrient digestion, detoxification, immunity, and mouthpart lubrication. Interestingly, several components of the larval saliva (e.g. apyrase and serine proteases) have also been reported to exist in adult female saliva, where they are though to help regulate a vertebrate hostâs immune response to bloodfeeding. In conclusion, our results suggest that the salivary glands are important components of both the digestive and immune systems of larval mosquitoes, and that their study might provide clues about the evolution of adaptations to bloodfeeding observed in adults. Experiment Overall Design: We attempted to identify transcripts that are both present and significantly enriched in the salivary glands (SG) of the 4th instar Anopheles gambiae larva. Transcripts were considered as present in the SG if the built-in Affymetrix detection call scored them as such in all three biological replicates of the experiment. Expression values (i.e. signal intensities) of transcripts from the salivary glands were compared to those of a matching group of whole 4th instar larvae using a t-test. Transcripts showing an expression value two-fold or higher (p < 0.01; false discovery rate=1%) in the SG as compared to whole larvae were considered to be significantly enriched.

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Transcriptomic Analyses of Secreted Proteins From the Salivary Glands of Wheat Midge Larvae.

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