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Sea anemones are sessile invertebrates belonging to the Cnidaria phylum and their survival and evolutive success is highly related to the ability of producing and fast inoculating venom. Sea anemones’ venom is enriched with biomolecules like polypeptides, proteins, polyamines and salts and some of these are potent toxins able to interact with sodium and potassium ion channels1,2, blocking or reducing their sensibility, leading to diverse effects on the prey, including fast inability to move. Toxins are intensely produced in tentacles and are also frequent in the mucus3. Bunodosoma caissarum is an endemic sea anemone specie which populates the intertidal zone of the Brazil’s southern coast. Some toxins were previously isolated like phospholipase A2, neurotoxins BcIII and IV and caissarolisin I. In order to bioprospect new sequences, omic approaches provide a detailed overview of biomolecules present in all kinds of samples, so some of them were used in this study. We performed a transcriptomic analysis of tentacles and peptidome and proteome of tentacles and mucus of B. caissarum. The assembly, annotation and quality assessment of the transcriptome were performed using Trinity RNA-Seq de novo, Trinotate and BUSCO, respectively. Peptides were extracted, measured by Fluorescamine and analyzed by liquid chromatography coupled to mass espectrometry (ESI-LC/MS/MS), with eletrospray ionization (ESI)4. Peptidome and proteome analysis was performed using PEAKS software. Putative toxin genes were identified with BLASTp local alignment between TransDecoder sequences and Toxprot database.Transcriptome assembly by Trinity resulted in 186.978 transcripts. Trinotate identified 23,444 annotated genes, 1% of them were characterized as toxins. BLASTp search resulted in 2055 genes with putative association with toxin activity, which were grouped in classes. Preliminary analysis from mucus and tentacle’s proteome revealed 746 proteins, 26% of them uncharacterized, and validated the presence of Potassium channel type 1 and 3, Sodium channel inhibitor and phospholipase A2 toxins. Differential protein expression analysis showed a higher abundance of known toxins in mucus than in tentacles samples. In the mucus, 118 proteins were found in higher expression levels, being 16 of them toxins, whilst in the tentacles, 316 proteins were highly expressed with 6 of them identified as toxins. Furthermore, new peptide sequences with similar structure to toxins were enriched in mucus peptidome. In conclusion, integrated omics improved the understanding about tentacle and mucus composition of specie.

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1	Maria Esquinca(eduarda.mazzi@unesp.br)
2	Leandro Castro(leandro.mantovani@unesp.br)

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1	Sato, K., Kadota, Y. & Shirasu, K (2019). Plant Immune Responses to Parasitic Nematodes. Front. Plant Sci. 10, 1–14. Abad, P. et al. (2008) Genome sequence of the metazoan plant-parasitic nematode Meloidogyne incognita. Nat. Biotechnol. 26, 909–915. Basso, M. F. et al. (2020) MiDaf16-like and MiSkn1-like gene families are reliable targets to develop biotechnological tools for the control and management of Meloidogyne incognita. Sci. Rep. 10, 1-13. Chaudhary, S., Dutta, T. K., Shivakumara, T. N. & Rao, U. (2019) RNAi of esophageal gland- specific gene Mi-msp-1 alters early stage infection behaviour of root-knot nematode, Meloidogyne incognita. J. Gen. Plant Pathol. 85, 232–242. Maule, A. G. et al. (2011) An eye on RNAi in nematode parasites. Trends Parasitol. 27, 505-513 . Perry R.N. & Moens M. (eds). 2013. Plant Nematology, Second edition. Wallingford, Oxfordshire, UK and Boston, USA, Russian Journal of Nematology, 2014, 22 (1), 77-82 Blanc-Mathieu, R. et al. (2017) Hybridization and polyploidy enable genomic plasticity without sex in the most devastating plant-parasitic nematodes. PLoS Genetics vol. 13.

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