The Evolution of Venom by Co-option of Single-Copy Genes Ellen O. Martinson, Mrinalini, Yogeshwar D. Kelkar, Ching-Ho Chang, John H. Werren  Current Biology  Volume 27, Issue 13, Pages 2007-2013.e8 (July 2017) DOI: 10.1016/j.cub.2017.05.032 Copyright © 2017 Elsevier Ltd Terms and Conditions

Figure 1 Venom Genes Are Rapidly Lost and Gained in the Specialized Venom Gland of Parasitoid Wasps (A) Species phylogeny of parasitoid wasps displaying gained (red) and lost (blue) venom genes. The total number of venom genes and the insect orders that the wasp parasitizes are shown for each species. Only venom genes that can be unambiguously assigned gain or loss along a phylogenetic branch are shown in the figure. See also Figures S1 and S2. (B) The number of genes that account for different percentages of mapped reads in the adult female (green), adult male (red), and venom gland (blue) transcriptomes of T. sarcophagae, showing gene expression specialization in the venom gland. (C) Cis-regulation of venom gland expression is revealed by comparing venom gland expression level of homologous venom genes in the parental species to allelic specific expression in venom glands of N. vitripennis × N. giraulti F1 hybrid females. The high correlation in expression is evidence that changes in gene expression between the species are due to cis-regulatory changes in the vicinity of the gene. Reciprocal cross and maternal effect analyses are available at FigShare (http://dx.doi.org/10.6084/m9.figshare.4731850). Current Biology 2017 27, 2007-2013.e8DOI: (10.1016/j.cub.2017.05.032) Copyright © 2017 Elsevier Ltd Terms and Conditions

Figure 2 Expression of Venom Genes in the Developmental Stages of N. vitripennis and T. sarcophagae Venom gene expression is shown in the venom gland and female and male whole bodies of T. sarcophagae and N. vitripennis for five developmental stages (adult, pupa, larva, late embryo, and early embryo) using boxplots. Expression in all T. sarcophagae developmental stages and the T. sarcophagae and N. vitripennis venom glands was measured in an RNA-seq analysis using log2(FPKM). Expression in N. vitripennis developmental stages was measured in a tiling array analysis using median log2(fluorescence/background intensity) of probes, in which any value above the dashed line is considered expressed relative to control oligos [31]. Expression level of shared (purple, n = 33) and newly recruited (yellow, T. sarcophagae n = 34, Nasonia clade n = 16) venom genes are compared to highly expressed ribosomal protein genes (gray boxes, n = 58). Bolded red branches in the phylogenies indicate the species in which the genes are venom genes. See also Figure S3. Current Biology 2017 27, 2007-2013.e8DOI: (10.1016/j.cub.2017.05.032) Copyright © 2017 Elsevier Ltd Terms and Conditions

Figure 3 Expression of Shared and Functionally Lost Venom Genes across Development Venom gene expression was measured in the venom gland and in female and male whole bodies for five developmental stages (adult, pupa, larva, late embryo, and early embryo). Expression in the N. vitripennis venom gland was measured in an RNA-seq analysis using log2(FPKM), and expression in the N. vitripennis developmental stages was measured in a tiling array analysis using median log2(fluorescence/background intensity) of probes, in which any value above the dashed line is considered expressed relative to control oligos. Bolded red branches on the phylogenies indicate in which species the genes are venom genes. (A) Venom genes shared between all four species (n = 33). (B) Genes that have lost venom status in N. vitripennis (n = 4). (C) Genes that have lost venom status in N. vitripennis and N. giraulti (n = 8). Analysis of lost venom expression in N. giraulti is not included because the requisite stage-specific expression data are not available. See also Table S3. Current Biology 2017 27, 2007-2013.e8DOI: (10.1016/j.cub.2017.05.032) Copyright © 2017 Elsevier Ltd Terms and Conditions