{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,25]],"date-time":"2026-10-25T05:04:29Z","timestamp":1792904669343,"version":"4.5.0"},"reference-count":77,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2019,11,1]],"date-time":"2019-11-01T00:00:00Z","timestamp":1572566400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005739","name":"Universidad Nacional Aut\u00f3noma de M\u00e9xico","doi-asserted-by":"publisher","award":["IN202619"],"award-info":[{"award-number":["IN202619"]}],"id":[{"id":"10.13039\/501100005739","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Scorpion toxins are thought to have originated from ancestral housekeeping genes that underwent diversification and neofunctionalization, as a result of positive selection. Our understanding of the evolutionary origin of these peptides is hindered by the patchiness of existing taxonomic sampling. While recent studies have shown phylogenetic inertia in some scorpion toxins at higher systematic levels, evolutionary dynamics of toxins among closely related taxa remain unexplored. In this study, we used new and previously published transcriptomic resources to assess evolutionary relationships of closely related scorpions from the family Hadruridae and their toxins. In addition, we surveyed the incidence of scorpine-like peptides (SLP, a type of potassium channel toxin), which were previously known from 21 scorpion species. We demonstrate that scorpine-like peptides exhibit gene duplications. Our molecular analyses demonstrate that only eight sites of two SLP copies found in scorpions are evolving under positive selection, with more sites evolving under negative selection, in contrast to previous findings. These results show evolutionary conservation in toxin diversity at shallow taxonomic scale.<\/jats:p>","DOI":"10.3390\/toxins11110637","type":"journal-article","created":{"date-parts":[[2019,11,1]],"date-time":"2019-11-01T12:30:50Z","timestamp":1572611450000},"page":"637","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Hadrurid Scorpion Toxins: Evolutionary Conservation and Selective Pressures"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6062-282X","authenticated-orcid":false,"given":"Carlos E.","family":"Santib\u00e1\u00f1ez-L\u00f3pez","sequence":"first","affiliation":[{"name":"Department of Biology, Eastern Connecticut State University, 83 Windham St. Willimantic, CT 06266, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew R.","family":"Graham","sequence":"additional","affiliation":[{"name":"Department of Biology, Eastern Connecticut State University, 83 Windham St. Willimantic, CT 06266, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prashant P.","family":"Sharma","sequence":"additional","affiliation":[{"name":"Department of Integrative Biology, University of Wisconsin-Madison, 430 Lincoln Drive, Madison, WI 53706, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8713-0224","authenticated-orcid":false,"given":"Ernesto","family":"Ortiz","sequence":"additional","affiliation":[{"name":"Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnolog\u00eda, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Avenida Universidad 2001, Cuernavaca, Morelos 62210, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6357-2661","authenticated-orcid":false,"given":"Lourival D.","family":"Possani","sequence":"additional","affiliation":[{"name":"Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnolog\u00eda, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Avenida Universidad 2001, Cuernavaca, Morelos 62210, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,11,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1146\/annurev.genom.9.081307.164356","article-title":"The Toxicogenomic Multiverse: Convergent Recruitment of Proteins Into Animal Venoms","volume":"10","author":"Fry","year":"2009","journal-title":"Annu. Rev. Genom. Hum. Genet."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1111\/brv.12062","article-title":"Poisons, toxungens, and venoms: Redefining and classifying toxic biological secretions and the organisms that employ them","volume":"89","author":"Nelsen","year":"2013","journal-title":"Biol. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1016\/j.toxicon.2012.11.022","article-title":"The venom optimization hypothesis revisited","volume":"63","author":"Morgenstern","year":"2013","journal-title":"Toxicon"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.tree.2012.10.020","article-title":"Complex cocktails: The evolutionary novelty of venoms","volume":"28","author":"Casewell","year":"2013","journal-title":"Trends Ecol. Evol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1146\/annurev-ento-120811-153650","article-title":"Spider-Venom Peptides: Structure, Pharmacology, and Potential for Control of Insect Pests","volume":"58","author":"King","year":"2013","journal-title":"Annu. Rev. Entomol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Torres, A.F.C., Huang, C., Chong, C.-M., Leung, S.W., Prieto-da-Silva, \u00c1.R.B., Havt, A., Quinet, Y.P., Martins, A.M.C., Lee, S.M.Y., and R\u00e1dis-Baptista, G. (2014). Transcriptome Analysis in Venom Gland of the Predatory Giant Ant Dinoponera quadriceps: Insights into the Polypeptide Toxin Arsenal of Hymenopterans. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0087556"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1146\/annurev-ento-120709-144849","article-title":"Venom Proteins from Endoparasitoid Wasps and Their Role in Host-Parasite Interactions","volume":"56","author":"Asgari","year":"2011","journal-title":"Annu. Rev. Entomol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1016\/j.toxicon.2011.01.004","article-title":"On the venom system of centipedes (Chilopoda), a neglected group of venomous animals","volume":"57","author":"Undheim","year":"2011","journal-title":"Toxicon"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2124","DOI":"10.1093\/molbev\/msu162","article-title":"Clawing through Evolution: Toxin Diversification and Convergence in the Ancient Lineage Chilopoda (Centipedes)","volume":"31","author":"Undheim","year":"2014","journal-title":"Mol. Biol. Evol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"679","DOI":"10.3390\/toxins7030679","article-title":"Centipede Venom: Recent Discoveries and Current State of Knowledge","volume":"7","author":"Undheim","year":"2015","journal-title":"Toxins"},{"key":"ref_11","first-page":"48","article-title":"The First Venomous Crustacean Revealed by Transcriptomics and Functional Morphology: Remipede Venom Glands Express a Unique Toxin Cocktail Dominated by Enzymes and a Neurotoxin","volume":"31","author":"Blanke","year":"2013","journal-title":"Mol. Biol. Evol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Gopalakrishnakone, P., Possani, L.D., Schwartz, E.F., and Rodriguez de la Vega, R.C. (2015). Scorpion Venoms, Springer.","DOI":"10.1007\/978-94-007-6404-0"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Gopalakrishnakone, P., Corzo, G., de Lima, M.E., and Diego-Garcia, E. (2016). Spider Venoms, Springer.","DOI":"10.1007\/978-94-007-6389-0"},{"key":"ref_14","first-page":"287","article-title":"Scorpion toxins specific for Na+-channels","volume":"264","author":"Possani","year":"1999","journal-title":"FEBS J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2456","DOI":"10.3390\/toxins5122456","article-title":"Evolution Stings: The Origin and Diversification of Scorpion Toxin Peptide Scaffolds","volume":"5","author":"Sunagar","year":"2013","journal-title":"Toxins"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.toxicon.2015.06.029","article-title":"Overview of the Knottin scorpion toxin-like peptides in scorpion venoms: Insights on their classification and evolution","volume":"107","author":"Possani","year":"2015","journal-title":"Toxicon"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2391","DOI":"10.1093\/molbev\/msn179","article-title":"Evolution of snake venom disintegrins by positive Darwinian selection","volume":"25","author":"Comas","year":"2008","journal-title":"Mol. Biol. Evol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2434","DOI":"10.1016\/j.cub.2016.07.038","article-title":"The deep origin and recent loss of venom toxin genes in rattlesnakes","volume":"26","author":"Dowell","year":"2016","journal-title":"Curr. Bio."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1093\/molbev\/msp310","article-title":"Positions under positive selection\u2014Key for selectivity and potency of scorpion alpha toxins","volume":"27","author":"Weinberger","year":"2010","journal-title":"Mol. Biol. Evol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1093\/gbe\/evv253","article-title":"Effects of gene duplication, positive selection, and shifts in gene expression on the evolution of the venom gland transcriptome in widow spiders","volume":"8","author":"Haney","year":"2016","journal-title":"Genome Biol. Evol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.toxicon.2017.05.002","article-title":"Venom gland transcriptomic and venom proteomic analyses of the scorpion Megacormus gertschi D\u00edaz-Najera, 1966 (Scorpiones: Euscorpiidae: Megacorminae)","volume":"133","author":"Zamudio","year":"2017","journal-title":"Toxicon"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Santib\u00e1\u00f1ez-L\u00f3pez, C., Cid-Uribe, J., Batista, C., Ortiz, E., and Possani, L. (2016). Venom Gland Transcriptomic and Proteomic Analyses of the Enigmatic Scorpion Superstitionia donensis (Scorpiones: Superstitioniidae), with Insights on the Evolution of Its Venom Components. Toxins, 8.","DOI":"10.3390\/toxins8120367"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Romero-Guti\u00e9rrez, M., Santib\u00e1\u00f1ez-L\u00f3pez, C., Jim\u00e9nez-Vargas, J., Batista, C., Ortiz, E., and Possani, L. (2018). Transcriptomic and Proteomic Analyses Reveal the Diversity of Venom Components from the Vaejovid Scorpion Serradigitus gertschi. Toxins, 10.","DOI":"10.3390\/toxins10090359"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Luna-Ram\u00edrez, K., Quintero-Hern\u00e1ndez, V., Ju\u00e1rez-Gonz\u00e1lez, V.R., and Possani, L.D. (2015). Whole Transcriptome of the Venom Gland from Urodacus yaschenkoi Scorpion. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0127883"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.toxicon.2016.11.261","article-title":"The first report on transcriptome analysis of the venom gland of Iranian scorpion, Hemiscorpius lepturus","volume":"125","author":"Khalaj","year":"2017","journal-title":"Toxicon"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.jprot.2016.08.004","article-title":"Transcriptomic analysis of the venom glands from the scorpion Hadogenes troglodytes revealed unique and extremely high diversity of the venom peptides","volume":"150","author":"Zhong","year":"2017","journal-title":"J. Proteom."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Santib\u00e1\u00f1ez-L\u00f3pez, C.E., Ojanguren-Affilastro, A., and Sharma, P.P. (2019). Another one bites the dust: Taxonomic sampling of a key genus in phylogenomic datasets reveals more non-monophyletic groups in traditional scorpion classification. Inv. Syst., in press.","DOI":"10.1071\/IS19033"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1111\/j.1476-5381.2009.00147.x","article-title":"Characterization of hadrucalcin, a peptide from Hadrurus gertschi scorpion venom with pharmacological activity on ryanodine receptors","volume":"157","author":"Schwartz","year":"2009","journal-title":"Br. J. Pharmacol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Schwartz, E.F., Diego-Garcia, E., Rodr\u00edguez de la Vega, R.C., and Possani, L.D. (2007). Transcriptome analysis of the venom gland of the Mexican scorpion Hadrurus gertschi (Arachnida: Scorpiones). BMC Genom., 8.","DOI":"10.1186\/1471-2164-8-119"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"13813","DOI":"10.1074\/jbc.M112.343996","article-title":"Hg1, Novel Peptide Inhibitor Specific for Kv1.3 Channels from First Scorpion Kunitz-type Potassium Channel Toxin Family","volume":"287","author":"Chen","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.toxicon.2017.01.014","article-title":"Venom-gland transcriptomics and venom proteomics of the black- back scorpion (Hadrurus spadix) reveal detectability challenges and an unexplored realm of animal toxin diversity","volume":"128","author":"Rokyta","year":"2017","journal-title":"Toxicon"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.peptides.2006.06.012","article-title":"Wide phylogenetic distribution of Scorpine and long-chain \u03b2-KTx-like peptides in scorpion venoms: Identification of \u201corphan\u201d components","volume":"28","author":"Schwartz","year":"2007","journal-title":"Peptides"},{"key":"ref_33","first-page":"187","article-title":"Cytolytic and K+ channel blocking activities of \u03b2-KTx and scorpine-like peptides purified from scorpion venoms","volume":"65","author":"Schwartz","year":"2007","journal-title":"CMLS Cell. Mol. Life Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s00018-004-4149-1","article-title":"The scorpine family of defensins: Gene structure, alternative polyadenylation and fold recognition","volume":"61","author":"Zhu","year":"2004","journal-title":"CMLS Cell. Mol. Life Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/S0014-5793(00)01384-3","article-title":"Scorpine, an anti-malaria and anti-bacterial agent purified from scorpion venom","volume":"471","author":"Conde","year":"2000","journal-title":"FEBS Lett."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e5902","DOI":"10.7717\/peerj.5902","article-title":"Integration of phylogenomics and molecular modeling reveals lineage-specific diversification of toxins in scorpions","volume":"6","author":"Kriebel","year":"2018","journal-title":"PeerJ"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.ympev.2018.01.003","article-title":"A revised dated phylogeny of scorpions\u2014Phylogenomic support for ancient divergence of the temperate Gondwanan family Bothriuridae","volume":"122","author":"Sharma","year":"2018","journal-title":"Mol. Phylogen. Evol."},{"key":"ref_38","first-page":"255","article-title":"Cladistic analysis of superfamily Iuroidea, with emphasis on subfamily Hadrurinae (Scorpiones: Iurida)","volume":"43","author":"Fet","year":"2008","journal-title":"Bol. Soc. Entomol. Aragonesa"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3897\/zookeys.59.546","article-title":"Further observations on scorpion genera Hadrurus and Hoffmannihadrurus (Scorpiones, Caraboctonidae)","volume":"59","author":"Soleglad","year":"2010","journal-title":"Zookeys"},{"key":"ref_40","first-page":"20142953","article-title":"Phylogenomic resolution of scorpions reveals multilevel discordance with morphological phylogenetic signal","volume":"282","author":"Sharma","year":"2015","journal-title":"Proc. Biol. Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1111\/j.1365-2699.2010.02431.x","article-title":"Evolutionary drivers of phylogeographical diversity in the highlands of Mexico: A case study of the Crotalus triseriatus species group of montane rattlesnakes","volume":"38","author":"Bryson","year":"2010","journal-title":"J. Biogeogr."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"10919","DOI":"10.1073\/pnas.0409127102","article-title":"Timing the origin and expansion of the Mexican tropical dry forest","volume":"102","author":"Becerra","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Ferrusquia-Villafranca, I., and Gonz\u00e1lez-Guzm\u00e1n, L.I. (2005). Northern Mexico\u2019s landscape, part II: The biotic setting across time. Biodiversity, Ecosystems, and Conservation in Northern Mexico, Oxford University Press.","DOI":"10.1093\/oso\/9780195156720.003.0003"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1002\/bies.201500165","article-title":"Toxin structures as evolutionary tools: Using conserved 3D folds to study the evolution of rapidly evolving peptides","volume":"38","author":"Undheim","year":"2016","journal-title":"BioEssays"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1111\/2041-210X.12534","article-title":"Fast and accurate detection of evolutionary shifts in Ornstein-Uhlenbeck models","volume":"7","author":"Khabbazian","year":"2016","journal-title":"Methods Ecol. Evol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1093\/sysbio\/syv043","article-title":"Detecting Adaptive Evolution in Phylogenetic Comparative Analysis Using the Ornstein\u2013Uhlenbeck Model","volume":"64","author":"Cressler","year":"2015","journal-title":"Syst. Biol."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Kriebel, R., Khabbazian, M., and Sytsma, K.J. (2017). A continuous morphological approach to study the evolution of pollen in a phylogenetic context: An example with the order Myrtales. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0187228"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Von Reumont, B.M. (2018). Studying smaller and neglected organisms in modern evolutionary venomics implementing rnaseq (transcriptomics)\u2014A critical guide. Toxins, 10.","DOI":"10.3390\/toxins10070292"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"15534","DOI":"10.1038\/s41598-018-33943-5","article-title":"Comparative venom-gland transcriptomics and venom proteomics of four sidewinder rattlesnake (Crotalus cerastes) lineages reveal little differential expression despite individual variation","volume":"8","author":"Hofmann","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1494","DOI":"10.1038\/nprot.2013.084","article-title":"De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis","volume":"8","author":"Haas","year":"2013","journal-title":"Nat. Protoc."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.toxicon.2018.06.085","article-title":"The diversity of venom components of the scorpion species Paravaejovis schwenkmeyeri (Scorpiones: Vaejovidae) revealed by transcriptome and proteome analyses","volume":"151","author":"Meneses","year":"2018","journal-title":"Toxicon"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"762","DOI":"10.1016\/j.celrep.2016.12.063","article-title":"A Tissue-Mapped Axolotl De Novo Transcriptome Enables Identification of Limb Regeneration Factors","volume":"18","author":"Bryant","year":"2017","journal-title":"Cell Rep."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.ympev.2019.02.021","article-title":"Phylogenomics facilitates stable scorpion systematics_ Reassessing the relationships of Vaejovidae and a new higher-level classification of Scorpiones (Arachnida)","volume":"135","author":"Monod","year":"2019","journal-title":"Mol. Phylogen. Evol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"2117","DOI":"10.1093\/molbev\/msw069","article-title":"A New Orthology Assessment Method for Phylogenomic Data: Unrooted Phylogenetic Orthology","volume":"33","author":"Ballesteros","year":"2016","journal-title":"Mol. Biol. Evol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1026","DOI":"10.1038\/nbt.3988","article-title":"MMseqs2 enables sensitive protein sequence searching for the analysis of massive data sets","volume":"35","author":"Steinegger","year":"2017","journal-title":"Nat. Biotech."},{"key":"ref_56","unstructured":"Dongen, S. (2000). Graph Clustering by Low Simulation. [Ph.D. Thesis, University of Utrecht]."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1575","DOI":"10.1093\/nar\/30.7.1575","article-title":"An efficient algorithm for large-scale detection of protein families","volume":"30","author":"Enright","year":"2002","journal-title":"Nucleic Acids Res."},{"key":"ref_58","first-page":"42","article-title":"Gnu parallel-the command-line power tool","volume":"36","author":"Tange","year":"2011","journal-title":"USENIX Mag."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"772","DOI":"10.1093\/molbev\/mst010","article-title":"MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability","volume":"30","author":"Katoh","year":"2013","journal-title":"Mol. Biol. Evol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1972","DOI":"10.1093\/bioinformatics\/btp348","article-title":"trimAl: A tool for automated alignment trimming in large-scale phylogenetic analyses","volume":"25","year":"2009","journal-title":"Bioinformatics"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1093\/molbev\/msu300","article-title":"IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies","volume":"32","author":"Nguyen","year":"2014","journal-title":"Mol. Biol. Evol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1093\/molbev\/msx281","article-title":"UFBoot2: Improving the ultrafast bootstrap approximation","volume":"35","author":"Hoang","year":"2018","journal-title":"Mol. Biol. Evol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"1188","DOI":"10.1093\/molbev\/mst024","article-title":"Ultrafast Approximation for Phylogenetic Bootstrap","volume":"30","author":"Minh","year":"2013","journal-title":"Mol. Biol. Evol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1038\/nmeth.4285","article-title":"ModelFinder: Fast model selection for accurate phylogenetic estimates","volume":"14","author":"Kalyaanamoorthy","year":"2017","journal-title":"Nat. Methods"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1093\/bioinformatics\/btv234","article-title":"ASTRAL-II: Coalescent-based species tree estimation with many hundreds of taxa and thousands of genes","volume":"31","author":"Mirarab","year":"2015","journal-title":"Bioinformatics"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1093\/oxfordjournals.molbev.a003974","article-title":"Estimating absolute rates of molecular evolution and divergence times: A penalized likelihood approach","volume":"19","author":"Sanderson","year":"2002","journal-title":"Mol. Biol. Evol."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1093\/bioinformatics\/btg412","article-title":"APE: Analyses of Phylogenetics and Evolution in R language","volume":"20","author":"Paradis","year":"2004","journal-title":"Bioinformatics"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1016\/j.ympev.2013.02.008","article-title":"Molecular dating of phylogenies by likelihood methods: A comparison of models and a new information criterion","volume":"67","author":"Paradis","year":"2013","journal-title":"Mol. Phylogen. Evol."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1071\/IS16078","article-title":"eadem figura manet: Measuring morphological convergence in diplocentrid scorpions (Arachnida: Scorpiones: Diplocentridae) under a multilocus phylogenetic framework","volume":"31","author":"Kriebel","year":"2017","journal-title":"Inv. Syst."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"D653","DOI":"10.1093\/nar\/gkq1058","article-title":"ArachnoServer 2.0, an updated online resource for spider toxin sequences and structures","volume":"39","author":"Herzig","year":"2010","journal-title":"Nucleic Acids Res."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1093\/molbev\/msn067","article-title":"An improved general amino acid replacement matrix","volume":"25","author":"Le","year":"2008","journal-title":"Mol. Biol. Evol."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1093\/sysbio\/sys029","article-title":"MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice Across a Large Model Space","volume":"61","author":"Ronquist","year":"2012","journal-title":"Syst. Biol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"W609","DOI":"10.1093\/nar\/gkl315","article-title":"PAL2NAL: Robust conversion of protein sequence alignments into the corresponding codon alignments","volume":"34","author":"Suyama","year":"2006","journal-title":"Nucleic Acids Res."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1093\/bioinformatics\/bti079","article-title":"HyPhy: Hypothesis testing using phylogenies","volume":"21","author":"Pond","year":"2005","journal-title":"Bioinformatics"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"1196","DOI":"10.1093\/molbev\/mst030","article-title":"FUBAR: A Fast, Unconstrained Bayesian AppRoximation for Inferring Selection","volume":"30","author":"Murrell","year":"2013","journal-title":"Mol. Biol. Evol."},{"key":"ref_76","doi-asserted-by":"crossref","unstructured":"Murrell, B., Wertheim, J.O., Moola, S., Weighill, T., Scheffler, K., and Kosakovsky Pond, S.L. (2012). Detecting individual sites subject to episodic diversifying selection. PLoS Genet., 8.","DOI":"10.1371\/journal.pgen.1002764"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"4","DOI":"10.32614\/RJ-2015-001","article-title":"Peptides: A package for data mining of antimicrobial peptides","volume":"7","author":"Osorio","year":"2015","journal-title":"R J."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/11\/11\/637\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:31:17Z","timestamp":1760189477000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/11\/11\/637"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,11,1]]},"references-count":77,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2019,11]]}},"alternative-id":["toxins11110637"],"URL":"https:\/\/doi.org\/10.3390\/toxins11110637","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,11,1]]}}}