Abstract
Introduction: Molecular divergence thresholds have been proposed to distinguish recently separated evolutive units, often displaying more accurate putative species assignments in taxonomic research compared to traditional morphological approaches. This makes DNA barcoding an attractive identification tool for a variety of marine invertebrates, especially for cryptic species complexes. Although GenBank and the Barcode of Life Data System (BOLD) are the major sequence repositories worldwide, very few have tested their performance in the identification of echinoderm sequences.
Objective: We use COI echinoderm sequences from local samples and the molecular identification platforms from GenBank and BOLD, in order to test their accuracy and reliability in the DNA barcoding identification for Central American shallow water echinoderms, at genus and species level.
Methods: We conducted sampling, tissue extraction, COI amplification, sequencing, and taxonomic identification for 475 specimens. The 348 obtained sequences were individually enquired with BLAST in GenBank as well as using the Identification System (IDS) in BOLD. Query sequences were classified depending on the best match result. McNemar’s chi-squared, Kruskal-Wallis’s and Mann-Whitney’s U tests were performed to prove differences between the results from both databases. Additionally, we recorded an updated list of species reported for the shallow waters of the Central American Pacific.
Results: We found 324 echinoderm species reported for Central American Pacific shallow waters. Only 118 and 110 were present in GenBank and BOLD databases respectively. We proposed 325 solved morphology-based identities and 21 provisional identifications in 50 putative taxa. GenBank retrieved 348 molecular-based identifications in 58 species, including twelve provisional identifications in tree taxa. BOLD recovered 170 COI identifications in 23 species with one provisional identification. Nevertheless, 178 sequences retrieved unmatched terms (in 34 morphology-based taxa). Only 86 sequences (25 %) were retrieved as correct identifications and 128 (37 %) as identification errors in both platforms. We include 84 sequences for eleven species not represented in GenBank and 65 sequences for ten species in BOLD Echinoderm COI databases. The identification accuracy using BLAST (175 correct and 152 incorrect identifications) was greater than with IDS engine (110 correct and 218 identification errors), therefore GenBank outperforms BOLD (Kruskal-Wallis = 41.625, df = 1, p < 0.001).
Conclusions: Additional echinoderm sample references are needed to improve the utility of the evaluated DNA barcoding identification tools. Identification discordances in both databases may obey specific parameters used in each search algorithm engine and the available sequences. We recommend the use of barcoding as a complementary identification source for Central American Pacific shallow water echinoderm species.
References
Altschul, S. F., Gish, W., Miller, W., Myers, E. W., & Lipman, D. J. (1990). Basic local alignment search tool. Journal of Molecular Biology, 215(3), 403–410. https://doi.org/10.1016/S0022-2836(05)80360-2
Alvarado, J. J., Barraza, E., & Sancho-Mejías, T. I. (2013). Central America Echinoderms: diversity, ecology and future perspectives. In J. J. Alvarado & F. A. Solís-Marín. (Eds.), Echinoderm research and diversity in Latin America (pp. 67–106). Springer. https://doi.org/10.1007/978-3-642-20051-9_3
Alvarado, J. J., Chacón-Monge, J. L., Solís-Marín, F. A., Pineda-Enríquez, T., Caballero-Ochoa, A. A., Rivera, S. S., & Chaves, R. R. (2017). Equinodermos del Museo de Zoología de la Universidad de Costa Rica. Revista de Biología Tropical, 65(S1), S272–S287. https://doi.org/10.15517/rbt.v65i1-1.31695
Alvarado, J. J., Chacón-Monge, J. L., Azofeifa-Solano, J. C., & Cortés, J. (2022). Diversity of deep-sea echinoderms from Costa Rica. Frontiers in Marine Science, 9, 918878. https://doi.org/10.3389/fmars.2022.918878
Alvarado, J. J., & Fabregat Malé, S. (2021). Echinoderm research perspectives: A Central American bibliometric review. Cuadernos de Investigación UNED, 13(2), e3535. http://dx.doi.org/10.22458/urj.v13i2.3535
Alvarado, J. J., & Solís-Marín, F. A. (2013). Echinoderm research and diversity in Latin America. In J. J. Alvarado & F. A. Solís-Marín (Eds.), Echinoderm research and diversity in Latin America (pp. 1–9). Springer. https://doi.org/10.1007/978-3-642-20051-9_1
Alvarado, J. J., Solís-Marín, F. A., & Ahearn, C. G. (2010). Echinoderm (Echinodermata) diversity in the Pacific coast of Central America. Marine Biodiversity, 40, 45–56. https://doi.org/10.1007/s12526-009-0032-5
Borrero-Pérez, G. H., & Vanegas-González, M. J. (2019). Holothuria (Mertensiothuria) viridiaurantia sp. nov. (Holothuriida, Holothuriidae), a new sea cucumber from the Eastern Pacific Ocean revealed by morphology and DNA barcoding. ZooKeys, 893, 1–19. https://doi.org/10.3897/zookeys.893.36013
Borrero-Pérez, H., & Vanegas-González, M. J. (2020). Riqueza, composición y distribución de los equinodermos de los Riscales y Morros del norte del Chocó, Pacífico Colombiano. In L. Chasqui (Ed.), Biodiversidad de los arrecifes rocosos (riscales y morros) del Pacífico Norte Chocoano (No. 116, pp. 232–282). Serie Publicaciones Generales de INVEMAR.
Chacón-Monge, J. L., Azofeifa-Solano, J. C., Alvarado, J. J., & Cortés, J. (2021). Área de Conservación Guanacaste echinoderms, North Pacific of Costa Rica. Revista de Biología Tropical, 69(S1), S487–S500. https://doi.org/10.15517/rbt.v69iSuppl.1.46391
Cortés, J., & Joyce, F. (2020). BioMar-ACG: A successful partnership to inventory and promulgate marine biodiversity. Biotropica, 52, 1104–1107. https://doi.org/10.1111/btp.12841
deWaard, J. R., Ivanova , N. V., Hajibabaei, M., & Hebert, P. D. N. (2008). Assembling DNA barcodes. Analytical protocols. Methods in Molecular Biology, 410, 275–293. https://doi.org/10.1007/978-1-59745-548-0_15
Ebach, M., Williams, D., & Morrone, J. (2006). Paraphyly is bad taxonomy. Taxon, 55(4), 831–832. https://doi.org/10.2307/25065678
Granja-Fernández, R., Herrero-Pérezrul, M., López-Pérez, R., Hernández, L., Rodríguez-Zaragoza, F., Wallace Jones, R., & Pineda-López, R. (2014). Ophiuroidea (Echinodermata) from coral reefs in the Mexican Pacific. ZooKeys, 406, 101–145. https://doi.org/10.3897/zookeys.406.6306
Granja-Fernández, R., Pineda-Enríquez, T., Solís-Marín, F. A. & Laguarda-Figueras, A. (2020). Ophioderma hendleri sp. nov. (Echinodermata: Ophiuroidea: Ophiodermatidae) and its congeners from the Eastern Pacific. European Journal of Taxonomy 729, 11–41. https://doi.org/10.5852/ejt.2020.729.1187
Grzywacz A, Wyborska D, & Piwczyński M. (2017). DNA barcoding allows identification of European Fanniidae (Diptera) of forensic interest. Forensic Science International, 278, 106–114. https://doi.org/10.1016/j.forsciint.2017.06.023
Hörandl, E. (2007). Neglecting evolution is bad taxonomy. Taxon, 56 (1), 1–5.
Ivanova, N. V., deWaard, J. R., & Hebert, P. D. N. (2006). An inexpensive, automation-friendly protocol for recovering high-quality DNA. Molecular Ecology Notes, 6(4), 998–1002. https://doi.org/10.1111/j.1471-8286.2006.01428.x
Kress, W. J. (2014). Valuing collections. Science, 346(6215), 1310. https://doi.org/10.1126/science.aaa4115
Laakmann, S., Boos, K., Knebelsberger, T., Raupach, M., & Neumann, H. (2016). Species identification of echinoderms from the North Sea by combining morphology and molecular data. Helgoland Marine Research, 70, 18. https://doi.org/10.1186/s10152-016-0468-5
Layton, K. K. S., Corstorphine, E. A., & Hebert, P. D. N. (2016). Exploring Canadian echinoderm diversity through DNA barcodes. PLoS ONE 11(11), e0166118. https://doi.org/doi:10.1371/journal.pone.0166118
Lessios, H. A. (2005). Echinoids of the Pacific waters of Panama: status of knowledge and new records. Revista de Biología Tropical, 53(S3), 147–170.
Martín-Cao-Romero, C., Solís-Marín, F. A., Laguarda-Figueras, A., & Buitrón-Sánchez. B. E. (2017). Phataria unifascialis (Valvatida: Ophidiasteridae) from the Eastern Pacific: Redescription and skeletal morphology. Revista de Biología Tropical, 65(S1), S258–S271. https://doi.org/10.15517/RBT.V65I1-1.31694
Massin, C., Zulfigar, Y., Tan Shau Hwai, A., & Rizzal, S. (2002). The genus Stichopus (Echinodermata: Holothuroidea) from the Johore Marine Park (Malaysia) with the description of two new species. Biologie, 72, 73–99.
Meiklejohn, K. A., Damaso, N., & Robertson, J. M. (2019). Assessment of BOLD and GenBank–Their accuracy and reliability for the identification of biological materials. PloS ONE, 14(6), e0217084. https://doi.org/10.1371/journal.pone.0217084
Miralles, A., Bruy, T., Wolcott, K., Scherrz, M., Begerow, D., Beszteri, B., Bonkowski, M., Fekden, J., Hemeinholzer, B., Glaw, F., Glöckner, F., Hawlitschek, O., Kostadinov, I., Nattkemper, T., Printzen, C., Renz, J., Rybalka, N., Stadler, M., Weibulat. T., … Vences, M. (2020). Repositories for taxonomic data: Where we are and what is missing. Systematic Biology, 69(6), 1231–1253. https://doi.org/10.1093/sysbio/syaa026
Ratnasingham, S., & Hebert, P. D. N. (2007). BOLD: The Barcode of Life Data System (www. barcodinglife. org). Molecular Ecology Notes, 7, 355–364. https://doi.org/10.1111/j.1471-8286.2007.01678.x
Solís-Marín, F. A., Alvarado, J. J., Abreu-Pérez, M., Aguilera, O., Alió, J., Bacallado-Aránega, J. J., Barraza, E., Benavides-Serrato, M., Benítez-Villalobos, F., Betancourt-Fernández, L., Borges, M., Brandt, M., Brogger, M. I., Helena Borrero-Pérez, G., Buitrón-Sánchez, B. E., Campos, L. S., Cantera, J., Clemente, S., Cohen-Renfijo, M., … Williams, S. M. (2013). Appendix. In J. J. Alvarado, & F. A. Solís-Marín (Eds.), Echinoderm Research and Diversity in Latin America (pp. 543–654). Springer.
Solís-Marín, F. A., Alvarado, J. J., Conejeros-Vargas, C., & Ochoa, A. A. (2020). Pentamera fonsecae n. sp. a new species of sea cucumber from the Pacific coast of Costa Rica (Holothuroidea: Dendrochirotida: Thyonidae). Zootaxa, 4878, 581–588. https://doi.org/10.11646/zootaxa.4878.3.9
Solís-Marín, F. A., Arriaga-Ochoa, J. A., Laguarda-Figueras, A., Frontana-Uribe, S. C., & Durán-González, A. (2009). Holothuroideos (Echinodermata: Holothuroidea) del Golfo de California, México [Informe técnico]. Comisión Nacional para el Conocimiento y Uso de la Biodiversidad e Instituto de Ciencias del Mar y Limnología de la Universidad Nacional Autónoma de México, México.
Solís-Marín, F. A., Laguarda-Figueras, A., Estrada-Rodríguez, P., Honey-Escandón, M., Cao-Romero, C., & Durán-Gonzáles, A. (2014). Los asteroideos (Echinodermata: Asteroidea) del Golfo de California, México [Informe técnico]. Instituto de Ciencias del Mar y Limnología de la Universidad Nacional Autónoma de México (UNAM), Secretaría de Medio Ambiente y Recursos Naturales (SEMARNAT) e Instituto Nacional de Ecología y Cambio Climático (INECC); México.
Sonet, G., Smitz, N., Vangestel, C., & Samyn, Y. (2022). DNA barcoding echinoderms from the East Coast of South Africa. The challenge to maintain DNA data connected with taxonomy. PLoS ONE, 17(10), e0270321. https://doi.org/10.1371/journal.pone.0270321
Schilthuizen, M., Vairappan, C., Slade, E., Mann, D., & Miller, J. (2015). Specimens as primary data: museums and ‘open science’. Trends in Ecology and Evolution, 30(5), 237–238. https://doi.org/10.1016/j.tree.2015.03.002
Varela-Sánchez, A., Templado, J., & Machordom, A. (2020). A. Morphological and molecular approaches to the study of ophiuroids (Echinodermata: Ophiuroidea) at Las Baulas Marine National Park (North Pacific, Costa Rica). Revista de Biología Tropical, 68(S3), 803–817. https://doi.org/10.15517/rbt.v68i3.39283
Woo, S. P., Zulfigar, Y., Tan, S. H., Kajihara, H., & Fujita, T. (2015). Sea cucumbers of the genus Stichopus Brandt, 1835 (Holothuroidea, Stichopodidae) in Straits of Malacca with description of a new species. ZooKeys, 545, 1–26. https://doi.org/10.3897/zookeys.545.6415
WoRMS Editorial Board (2024). World Register of Marine Species. Vlaams Instituut voor de Zee. https://doi.org/10.14284/170
Comments
This work is licensed under a Creative Commons Attribution 4.0 International License.