Co-option of wing-patterning genes underlies the evolution of the treehopper helmet

Abstract

Understanding the origin of novelty is a key question in evolutionary developmental biology. In arthropods, the body wall has served as a repeated source of morphological novelty. In treehoppers, an ancestrally flat part of the dorsal body wall (the pronotum) was transformed into a three-dimensional structure (the helmet), which was subsequently moulded by natural selection into diverse shapes. Here, we test three hypotheses for the developmental origin of the helmet by comparing body-region transcriptomes in a treehopper and a leafhopper that retains more ancestral morphology. In leafhoppers, pronotal gene expression is most similar to that of its serial homologue, the mesonotum. By contrast, in treehoppers, helmet gene expression is most similar to that of wings, supporting the wing-patterning network co-option hypothesis for the origin of the helmet. These results suggest that serial homologues may diverge evolutionarily through replacement of, rather than tinkering with, their ancestrally shared patterning network.

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Fig. 1: Pronotal morphology in treehoppers and leafhoppers.
Fig. 2: Character trees showing predictions and results of hierarchical clustering based on differential gene expression.
Fig. 3: PCA of genes differentially expressed across body regions.
Fig. 4: Character trees based on hierarchical clustering of annotated subsets of differentially expressed genes.
Fig. 5: GO term enrichment for differentially expressed genes upregulated in the wings and pronotum.
Fig. 6: Expression of candidate genes related to wing development across body regions.

Data availability

The raw sequencing reads and refined transcriptome assemblies that form the basis for this study have been deposited in the NCBI Sequence Reads Archive (SRA) and Transcriptome Shotgun Assembly (TSA) database, respectively. All data are components of NCBI BioProject no. PRJNA415461. The transcriptome assembly for H. vitripennis has been deposited at DDBJ/ENA/GenBank under the accession GHXA00000000. The version described in this paper is the first version, GHXA01000000. The transcriptome assembly for E. carinata has been deposited at DDBJ/ENA/GenBank under the accession GHWZ00000000. The version described in this paper is the first version, GHWZ01000000. Raw reads are attached to SRA study no. SRP152991 with accession codes SRR9942929SRR9942973.

Code availability

Analytical code, along with test data, has been posted at www.github.com/fishercera/TreehopperSeq.

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Acknowledgements

Financial support for this work was provided by the National Science Foundation (no. IOS 1656572) to E.L.J. C.R.F. was supported in part by an Outstanding Scholar Fellowship from the University of Connecticut. Seed funding for pilot studies was provided to C.R.F. by the Sigma Xi Grants in Aid of Research programme, the Society for the Study of Evolution Rosemary Grant Award programme and the Connecticut Museum of Natural History. Analyses were run on the UConn Health Center High Performance Computing cluster, administered by the Computational Biology Core within the Institute for Systems Genomics. We appreciate assistance with treehopper colony maintenance provided by M. Opel and other UConn EEB greenhouse staff, A. Chiu and M. Deering. Support for library quantification and fragment analysis was provided by B. Reese and L. Li at the UConn Center for Genome Innovation. Valuable comments on the manuscript were provided by C. Henry, C. Schlichting, F. Smith and Y. Yuan. We acknowledge the assistance of Y. Son, of the California Department of Food and Agriculture Pierce’s Disease Control Program, for providing H. vitripennis specimens. We thank P. Coin, P. Kirillov, M. Schmidt and K. Swing for granting permission to use their photographs.

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C.R.F. and E.L.J. conceived the study and designed the sampling. All authors contributed to the data analysis design. C.R.F. reared specimens, prepared libraries, analysed the data in consultation with E.L.J. and J.L.W. and wrote the first draft of the manuscript. C.R.F., E.L.J. and J.L.W. revised the manuscript.

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Correspondence to Cera R. Fisher.

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Supplementary Methods 1 and 2, Supplementary Figures 1–9 and Supplementary Tables 1–5.

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Supplementary Data

Identifiers and annotations for transcripts upregulated in Entylia pronotum, Homalodisca pronotum and, where transcript is a single-copy orthologue, whether it is upregulated in the other species’ pronotum.

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Fisher, C.R., Wegrzyn, J.L. & Jockusch, E.L. Co-option of wing-patterning genes underlies the evolution of the treehopper helmet. Nat Ecol Evol 4, 250–260 (2020). https://doi.org/10.1038/s41559-019-1054-4

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