Multiple stages of development (mature and early stages with and without spores, respectively) also characterised the pooled myxoworms collected from bryozoan hosts

Multiple stages of development (mature and early stages with and without spores, respectively) also characterised the pooled myxoworms collected from bryozoan hosts. Our collective approaches enable us to test the following hypotheses: (H1) venom toxins have been conserved and repurposed in endoparasitic cnidarians; (H2) toxins of endoparasitic cnidarians are more divergent from homologs in free-living relatives; and (H3) toxin diversity will be reduced in myxozoans in keeping with the general trend for reduction in genome size in this group. version 3.2 based on amino acid sequences. Numbers in the Nitrarine 2HCl nodes show posterior probability values. peerj-09-11208-s009.pdf (59K) DOI:?10.7717/peerj.11208/supp-9 Supplemental Information 10: Phylogenetic tree of the Phospholipase A2 gene family. The tree was constructed with Bayesian inference using MrBayes version 3.2 based on amino acid sequences. Numbers in the nodes show posterior probability values. peerj-09-11208-s010.pdf (4.7K) DOI:?10.7717/peerj.11208/supp-10 Supplemental Information 11: Phylogenetic tree of the Kunitz mature domain constructed with maximum likelihood (ML) method using IQ-TREE version 2.1.1 software. This tree was rooted with two plants sequences from I3A kunitz family. peerj-09-11208-s011.pdf (4.0M) DOI:?10.7717/peerj.11208/supp-11 Supplemental Information 12: Phylogenetic Tree of the Kunitz-type venom gene family showing the un-condensed clades. peerj-09-11208-s012.pdf (2.8M) DOI:?10.7717/peerj.11208/supp-12 Supplemental Information 13: Kunitz mature domain sequences of validated proteins that contain the Kunitz domain used in the analysis in FASTA format. peerj-09-11208-s013.fasta (111K) DOI:?10.7717/peerj.11208/supp-13 Supplemental Information 14: Presence/absence matrix of toxin protein families in major cnidarian classes. peerj-09-11208-s014.xlsx (6.5K) DOI:?10.7717/peerj.11208/supp-14 Supplemental Information 15: Accession numbers for UNIPROT/genbank Kunitz sequences. peerj-09-11208-s015.docx (147K) DOI:?10.7717/peerj.11208/supp-15 Nitrarine 2HCl Supplemental Information 16: Accession numbers for UNIPROT/genbank PLA2 sequences. peerj-09-11208-s016.docx (18K) DOI:?10.7717/peerj.11208/supp-16 Supplemental Information 17: ELISA measurements of specificity and sensitivity of the polyclonal antibodies raised against various putative toxins. peerj-09-11208-s017.xlsx (15K) DOI:?10.7717/peerj.11208/supp-17 Supplemental Information 18: Transcriptome sequencing and assembly statistics. peerj-09-11208-s018.docx (14K) DOI:?10.7717/peerj.11208/supp-18 Supplemental Information 19: Putative venom toxins identified from the transcriptome and proteome of substantiates toxin translation and thus illustrates a repurposing of toxin function for endoparasite development and interactions with hosts, rather than for prey capture or defence. Whether myxozoan venom candidates are expressed in transmission stages (e.g. in nematocysts or secretory vesicles) requires further investigation. as sister to Myxozoa, forming the Endocnidozoa (Chang et al., 2015; Kayal et al., 2018) (Fig. 1). Myxozoans comprise some 20% of currently recognised cnidarian species (Okamura, Hartigan & Naldoni, 2018) and exploit vertebrate and invertebrate hosts in complex life cycles. Extremely rapid rates of molecular evolution, morphological simplification and miniaturisation long precluded higher-level placement of myxozoans (Okamura, Gruhl & Reft, 2015) and may be associated with their greatly reduced genome sizes (Chang et al., 2015). Myxozoans are comprised of the sister taxa, Malacosporea and Myxosporea. The former possess tissues and develop into active vermiform myxoworms (e.g. entails a parasitic larval stage in eggs of paddlefish and sturgeon and a free-living, actively feeding adult stage Nitrarine 2HCl (Raikova, 1994). Open in a separate window Figure 1 Cladogram of Cnidaria displaying some major lineages including placement of species used in this study (greyscale clades).Phylogenetic tree representing some of the major clades of Cnidaria (based (Chang et al., 2015; Kayal et al., 2018)). The species sampled in this venomics study were the myxozoans and and the free-living staurozoan but did not address whether such putative toxins were distinct from those with nontoxic physiological functions. Proteomic analysis of isolated polar capsules from the myxozoan found minimal evidence for proteins similar to toxins of cnidarians or other venomous animals (a toxin-like ShK domain was identified in one peptide sequence, but this was later discounted (Piriatinskiy et al., 2017)). However, nematocysts and filament function have been shown to be highly variable in myxozoans (Ben-David et al., 2016). The Nitrarine 2HCl apparent loss of venom toxins in nematocysts of could be attributed to a structural adaptation that facilitates host attachment rather than toxin delivery. Here we tackle the question of whether endoparasites employ venom Rabbit Polyclonal to MAST4 toxins inherited from their free-living ancestors by presenting comparative transcriptomics, targeted proteomics and further expression studies on cnidarian taxa with entirely parasitic life cycles (and the myxosporeans and and were.