Abstract
The small size (< 5 mm) of marine microfilamentous green algae makes it difficult to identify based on field collections, remaining largely unnoticed. To catalogue the diversity of these diminutive marine algae, a combination of culture-based taxonomic studies and DNA barcoding has proven to be a successful approach. During subtidal surveys in the northeastern Brazil, microfilamentous green algae were isolated as epiphytes on red algae cultured in laboratory. In this work, we used thallus ontogeny and plastid tufA gene sequences to elucidate the taxonomic identity of these isolates. Molecular and morphological evidence revealed that Brazilian specimens correspond to Ulvella endozoica, a poorly known species thought to be endemic to Florida, its type locality. Since U. endozoica was not found in gorgonian corals, as in the original description, this species does not follow a host specificity pattern, growing in more diverse substrata than previously thought. The origin of U. endozoica in Brazil remains undetermined, and we concluded that it is a cryptogenic species. Limited sampling, troublesome taxonomy, and difficulty in finding Ulvella species make them overlooked in Western Atlantic, resulting in taxonomic and biogeographic gaps. Our results expand southward the known distribution range of U. endozoica, demonstrating that an integrative barcode and laboratory culture-based approach is critical to uncover these inconspicuous algae.
Similar content being viewed by others
References
Akaike H (1974) A new look at the statistical model identification. IEE Trans Automat Contr 19:716–723
Barata D, Fujii MT (2006) Ctenocladaceae e Ulvellaceae (Chlorophyta) do Espírito Santo e litoral norte de São Paulo, Brasil. Hoehnea 33:359–370
Benson DA, Cavanaugh M, Clark K, Karsch-Mizrachi I, Ostell J, Pruitt KD, Sayers EW (2018) GenBank. Nucleic Acids Res 46:41–47
Bernard M, Strittmatter M, Murúa P, Heesch S, Cho GY, Leblanc C, Peters AF (2019) Diversity, biogeography and host specificity of kelp endophytes with a focus on the genera Laminarionema and Laminariocolax (Ectocarpales, Phaeophyceae). Eur J Phycol 54:39–51
Bown P, Plumb J, Sánchez-Baracaldo P, Hayes PK, Brodie J (2003) Sequence heterogeneity of green (Chlorophyta) endophytic algae associated with a population of Chondrus crispus (Gigartinaceae, Rhodophyta). Eur J Phycol 38:153–163
Carlton JT (1996) Biological invasions and cryptogenic species. Ecology 77:1653–1655
Chapman ARO (1986) Population and community ecology of seaweeds. In: Blaxter JHS, Southwood AJ (eds) Advances in marine biology. Academic Press, London, pp 1–161
Cordeiro R, McFadden C, van Ofwegen L, Williams G (2020) World list of Octocorallia. Pseudoplexaura Wright & Studer, 1889. http://www.marinespecies.org/aphia.php?p=taxdetails&id=267779. Accessed on 18 October 2020
Díaz-Tapia P, Bárbara I, Cremades J, Verbruggen H, Maggs CA (2017) Three new cryptogenic species in the tribes Polysiphonieae and Streblocladieae (Rhodomelaceae, Rhodophyta). Phycologia 56:605–623
Díaz-Tapia P, Baldock L, Maggs CA (2020) Discovery of Flabellia petiolata (Halimedaceae, Chlorophyta) in the southern British Isles: a relict population or a new introduction. Aquat Bot 160:103160
Famà P, Wysor B, Kooistra WHCB, Zuccarello GC (2002) Molecular phylogeny of the genus Caulerpa (Caulerpales, Chlorophyta) inferred from chloroplast tufA gene. J Phycol 38:1040–1050
Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791
Geoffroy A, Destombe C, Kim B, Mauger S, Raffo MP, Kim MS, Le Gall L (2016) Patterns of genetic diversity of the cryptogenic red alga Polysiphonia morrowii (Ceramiales, Rhodophyta) suggest multiple origins of the Atlantic populations. Ecol Evol 6:5635–5647
Goldberg WM, Makemson JC, Colley SB (1984) Entocladia endozoica sp. nov., a pathogenic chlorophyte: structure, life history, physiology, and effect on its coral host. Biol Bull 166:368–383
Gomoiu MT, Alexandrov B, Shadrin N, Zaitsev Y (2002) The Black Sea – a recipient, donor and transit area for alien species. In: Leppäkoski E, Gollasch S, Olenin S (eds) Invasive aquatic species of Europe. Distribution, impacts and management. Springer, Dordrecht, pp 341–350
Guimarães-Costa AJ, Machado FS, Oliveira RRS, Silva-Costa V, Andrade MC, Giarrizzo T, Saint-Paul U, Sampaio I, Schneider H (2019) Fish diversity of the largest deltaic formation in the Americas – a description of the fish fauna of the Parnaíba Delta using DNA barcoding. Sci Rep 9:7530
Guiry MD, Guiry GM (2020) AlgaeBase. Worldwide electronic publication, National University of Ireland, Galway. http://www.algaebase.org. Accessed on 09 October 2020
Gunnarsson K, Nielsen R (2016) Culture and field studies of Ulvellaceae and other microfilamentous green seaweeds in subarctic and arctic waters around Iceland. Nova Hedwigia 103:17–46
Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98
Hansen GI, Hanyuda T, Kawai H (2018) Invasion threat of benthic marine algae arriving on Japanese tsunami marine debris in Oregon and Washington, USA. Phycologia 57:641–658
Hansen GI, West JA, Yoon HS, Goodman CD, Goër SL, Zuccarello GC (2019) Viator vitreocola gen. et sp. nov. (Stylonematophyceae), a new red alga on drift glass debris in Oregon and Washington, USA. Algae 34:71–90
Hewitt CL, Campbell ML, Thresher RE, Martin RB, Boyd S, Cohen BF, Currie DR, Gomon MF, Keough MJ, Lewis JA, Lockett MM, Mays N, McArthur MA, O’Hara TD, Poore GCB, Ross DJ, Storey MJ, Watson JE, Wilson RS (2004) Introduced and cryptogenic species in Port Phillip Bay, Victoria, Australia. Mar Biol 144:183–202
Hoffmann AJ, Santelices B (1991) Banks of algal microscopic forms: hypotheses on their functioning and comparisons with seed banks. Mar Ecol Prog Ser 79:185–194
Joly AB, Cordeiro M (1963) Two new species of Acrochaetium from southern Brazil. Bol Fac Fil Ciênc Let Univ S Paulo 19:135–146
Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 35:1547–1549
Leliaert F, Rueness J, Boedeker C, Maggs CA, Cocquyt E, Verbruggen H, De Clerck O (2009) Systematics of the marine microfilamentous green algae Uronema curvatum and Urospora microscopica (Chlorophyta). Eur J Phycol 44:487–496
Lewin J (1966) Silicon metabolism in diatoms. V. Germanium dioxide, a specific inhibitor of diatom growth. Phycologia 6:1–12
Maggs CA, Callow ME (2002) Algal spores. In: Encyclopedia of life sciences. Macmillan Publishers Ltd., Nature Publishing Group, London, pp 1–6
Marcelino VR, Verbruggen H (2016) Multi-marker metabarcoding of coral skeletons reveals a rich microbiome and diverse evolutionary origins of endolithic algae. Sci Rep 6:31508
Nielsen R, Petersen G, Seberg O, Daugbjerg N, O’Kelly CJ, Wysor B (2013) Revision of the genus Ulvella (Ulvellaceae, Ulvophyceae) based on morphology and tufA gene sequences of species in culture, with Acrochaete and Pringsheimiella placed in synonymy. Phycologia 52:37–56
Nielsen R, Gunnarsson K, Daugbjerg N, Petersen G (2014) Description of Ulvella elegans sp. nov. and U. islandica sp. nov. (Ulvellaceae, Ulvophyceae) from Iceland – a study based on morphology of species in culture and tufa gene sequences. Eur J Phycol 49:60–67
O’Kelly CJ, Wysor B, Bellows WK (2004) Gene sequence diversity and the phylogenetic position of algae assigned to the genera Phaeophila and Ochlochaete (Ulvophyceae, Chlorophyta). J Phycol 40:789–799
Oliveira EC, Paula EJ, Plastino EM, Petti R (1995) Metodologias para o cultivo axênico de macroalgas marinas in vitro. In: Alveal K, Ferrario ME, Oliveira EC, Sar E (eds) Manual de Métodos Ficológicos. Universidade de Concepción, Concepción, pp 429–455
Peters AF, Couceiro L, Tsiamis K, Küpper FC, Valero M (2015) Barcoding of cryptic stages of marine brown algae isolated from incubated substratum reveals high diversity in Acinestoporaceae (Ectocarpales, Phaeophyceae). Cryptogamie, Algol 36:3–29
Piñeiro-Corbeira C, Verbruggen H, Díaz-Tapia P (2019) Molecular survey of the red algal family Rhodomelaceae (Ceramiales, Rhodophyta) in Australia reveals new introduced species. J Appl Phycol 32:2535–2547
Posada D, Crandall KA (1998) MODELTEST: testing the model of DNA substitution. Bioinformatics 14:817–818
Rambaut A (2018) FigTree v1.4.2, a graphical viewer of phylogenetic trees. http://tree.bio.ed.ac.uk/software/figtree/. Accessed on 20 May 2020
Rinkel BE, Hayes P, Gueidan C, Brodie J (2012) A molecular phylogeny of Acrochaete and other endophytic green algae (Ulvales, Chlorophyta). J Phycol 48:1020–1027
Santelices B, Aedo D, Hoffmann A (2002) Banks of microscopic forms and survival to darkness of propagules and microscopic stages of macroalgae. Rev Chil Hist Nat 75:547–555
Saunders GW, Kucera H (2010) An evaluation of rbcL, tufA, UPA, LSU and ITS as DNA barcode markers for the marine green macroalgae. Cryptogamie, Algol 31:487–528
Saunders GW, McDevit DC (2012) Methods for DNA barcoding photosynthetic protists emphasizing the macroalgae and diatoms. In: Kress WJ, Erickson DL (eds) DNA barcodes: methods and protocols, vol 858. Methods Mol Biol, pp 207–222
Silva AGA, Stattegger K, Schwarzer K, Vital H, Heise B (2015) The influence of climatic variations on river delta hydrodynamics and morphodynamics in the Parnaíba Delta, Brazil. J Coast Res 314:930–940
Soares LP, Fujii MT (2020) Molecular assessment of flat Cystocloniaceae (Gigartinales, Rhodophyta) from Brazil with reinstatement of Calliblepharis jolyi and a new record of C. saidana for the Atlantic Ocean. Phytotaxa 439:243–254
Soares LP, Guimarães SMPB, Fujii MT, Yoneshigue-Valentin Y, Batista MGS, Yokoya NS (2019) Thallus ontogeny, morphology and molecular phylogeny of Madagascaria atlantica sp. nov. (Compsopogonophyceae, Rhodophyta), a diminutive crustose species uncovered in the Atlantic Ocean. Aquat Bot 159:103152
Soares LP, Guimarães SMPB, Fujii MT, Yoneshigue-Valentin Y, Batista MGS, Yokoya NS (2020) Rhodachlya westii sp. nov. (Rhodachlyales, Rhodophyta), a new species from Brazil revealed by an integrative taxonomic approach. Phycologia 59:346–354
Spalding MD, Fox HE, Allen GR, Davidson N, Ferdaña ZA, Finlayson M, Halpern BS, Jorge MA, Lombana A, Lourie SA, Martin KD, McManus E, Molnar J, Recchia CA, Robertson J (2007) Marine ecoregions of the world: a bioregionalization of coastal and shelf areas. Bioscience 57:573–583
Stamatakis A (2014) RaxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30:1312–1313
Thiers B (2020) Index Herbariorum: a global directory of public herbaria and associated staff. New York Botanical Garden’s Virtual Herbarium. http://sweetgum.nybg.org/science/ih/. Accessed on 20 March 2020
West JA, Hansen GI, Hanyuda T, Zuccarello GC (2016) Flora of drift plastics: a new red algal genus, Tsunamia transpacifica (Stylonematophyceae) from Japanese tsunami debris in the northeast Pacific Ocean. Algae 31:289–301
Wynne MJ (2017) A checklist of benthic marine algae of the tropical and subtropical Western Atlantic: fourth revision. Nova Hedwigia 145:1–202
Yarish C (1976) Polymorphism of selected marine Chaetophoraceae (Chlorophyta). Br Phycol J 11:29–38
Yokoya NS (2000) Apical callus formation and plant regeneration controlled by plant growth regulators on axenic cultures of the red alga Gracilariopsis tenuifrons (Gracilariales, Rhodophyta). Phycol Res 48:133–142
Acknowledgments
NSY, MTF, and YYV thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for the Productivity Fellowship (312355/2019-0, 304899/2017-8, and 301938/2019-9, respectively). LPS thanks the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Bolsista CAPES/BRASIL 88887.515974/2020-00). We thank the reviewers for their comments that improved the manuscript.
Funding
This study was partially funded by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES/AUXPE-CIMAR 1991/2014/23038.001431/2014-75) and the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP 2016/50370-7).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
Not applicable.
Sampling and field studies
All necessary permits for sampling were obtained by the authors from the competent authorities and are mentioned in the acknowledgements, when applicable.
Data availability
Data are available in herbarium SP and GenBank.
Author’s contribution
NSY, MTF, MGSB, and YYV conceived the study and collected the examined samples. SMPBG and NSY carried out the culture studies. LPS wrote the manuscript and performed the molecular analysis. LPS and SMPBG studied the morphological characters. All authors revised and approved the manuscript.
Additional information
Communicated by B. Beszteri
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Soares, L.P., Guimarães, S.M.P.B., Fujii, M.T. et al. New insights on the distribution and habitat of Ulvella endozoica (Ulvellaceae, Chlorophyta) in the tropical Southwestern Atlantic, based on thallus ontogeny in culture and DNA barcoding. Mar. Biodivers. 51, 12 (2021). https://doi.org/10.1007/s12526-020-01153-w
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s12526-020-01153-w