New Zealand Journal of Botany ISSN: 0028-825X (Print) 1175-8643 (Online) Journal homepage: http://www.tandfonline.com/loi/tnzb20 Molecular evidence of the presence of Dictyota dichotoma (Dictyotales: Phaeophyceae) in Argentina based on sequences from mtDNA and cpDNA and a discussion of its possible origin Erick Alves Pereira Lopes-Filho, Fabiano Salgueiro, Silvia Mattos Nascimento, M. Cecilia Gauna, Elisa R. Parodi & Joel Campos De Paula To cite this article: Erick Alves Pereira Lopes-Filho, Fabiano Salgueiro, Silvia Mattos Nascimento, M. Cecilia Gauna, Elisa R. Parodi & Joel Campos De Paula (2017) Molecular evidence of the presence of Dictyota�dichotoma (Dictyotales: Phaeophyceae) in Argentina based on sequences from mtDNA and cpDNA and a discussion of its possible origin, New Zealand Journal of Botany, 55:3, 293-305, DOI: 10.1080/0028825X.2017.1326387 To link to this article: https://doi.org/10.1080/0028825X.2017.1326387 View supplementary material Published online: 05 Jun 2017. Submit your article to this journal Article views: 181 View Crossmark data Citing articles: 1 View citing articles Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=tnzb20 NEW ZEALAND JOURNAL OF BOTANY, 2017 VOL. 55, NO. 3, 293–305 https://doi.org/10.1080/0028825X.2017.1326387 RESEARCH ARTICLE Molecular evidence of the presence of Dictyota dichotoma (Dictyotales: Phaeophyceae) in Argentina based on sequences from mtDNA and cpDNA and a discussion of its possible origin Erick Alves Pereira Lopes-Filhoa, Fabiano Salgueiroa, Silvia Mattos Nascimentoa, M. Cecilia Gaunab, Elisa R. Parodib and Joel Campos De Paula a a Programa de Pós-Graduação em Biodiversidade Neotropical, Universidade Federal do Estado do Rio de Janeiro, Rio de Janeiro, Brazil; bCONICET- Instituto Argentino de Oceanografía, Universidad Nacional del Sur, Bahía Blanca, Argentina ABSTRACT ARTICLE HISTORY Dictyota is a brown algae genus inhabiting tropical to warm temperate environments where it is an important food source, shelter and substrate to several species of invertebrates and other algae. The taxonomy of this genus is troublesome, with poor species delimitation and doubtful records in the literature. Dictyota dichotoma, the type species of the genus, was commonly accepted as a cosmopolitan species because of the inaccurate geographical distributional range as a consequence of misidentification in several parts of the world. Recent studies with molecular data revealed new species and a new understanding of the evolution and biogeography of the genus Dictyota. These studies confined natural populations of D. dichotoma to Europe (Atlantic and Mediterranean coasts) and Macaronesian islands (Azores, Madeira and Canary archipelagos). Also, they confirmed its presence in South Africa, but whether the species was native or introduced in South Africa could not be verified. In the present study two regions of cpDNA (psbA, rbcL) and one region of mtDNA (nad1) from Argentinian samples, identified morphologically as D. dichotoma, were analysed and compared to other Dictyota species. The identity of these samples as D. dichotoma was confirmed. A haplotype network analysis using all available psbA sequences distinguished seven haplotypes divided into two geographic groups: Atlantic–Mediterranean and Canarian. In Argentina and South Africa only the most common haplotype of the Atlantic–Mediterranean group was observed. According to the paleoceanographic currents and the presence of a European haplotype, the introduced nature of D. dichotoma is indicated. Received 14 February 2017 Accepted 1 May 2017 KEYWORDS Dictyotaceae; exotic species; nad1; psbA; rbcL; South Atlantic Ocean CONTACT Joel Campos De Paula depaula.joelc@gmail.com Supplementary data available online at www.tandfonline.com/10.1080/0028825X.2017.1326387 © 2017 The Royal Society of New Zealand 294 E. A. P. LOPES-FILHO ET AL. Introduction Dictyota J.V. Lamouroux is an important component of the marine flora in tropical and subtropical environments and is commonly found from the lower range of the intertidal zone to deeper areas (c. 80 m) of the subtidal zone (Littler & Littler 2000; De Clerck et al. 2006; Bittner et al. 2008). The genus has attracted the attention of researchers for several reasons, including the production of bioactive metabolites (Vallim et al. 2005; De Paula et al. 2011) and its ecological role, such as the high biomass that may be used as a food source, shelter and that serves as a substrate to several species of invertebrates and other algae (Genzano & Rodríguez 1998; Stachowicz & Hay 2000; Gauna et al. 2015; Moore & Eastman 2015). The identification of Dictyota species may be challenging due to phenotypic plasticity, simple morphology and poor delineation among species, which is the reason for the description of hundreds of names for new taxa and doubtful geographical distributional ranges (Schnetter et al. 1987; De Clerck 2003; Tronholm et al. 2010a, 2010b; Gauna et al. 2013). In the southwest Atlantic, the genus Dictyota remains understudied and the species recorded have never been the subject of a formal review, although there have been some nomenclatural corrections (Oliveira Filho 1977; Széchy & De Paula 2016). Dictyota dichotoma (Hudson) J.V. Lamouroux was described as Ulva dichotoma in England in 1762 and later it was transferred to Dictyota in 1809 (Lamouroux 1809). During the 19th and 20th centuries, D. dichotoma was considered to be widely distributed from tropical to warm temperate areas of the world (Hwang et al. 2005; Tronholm et al. 2008, 2010b) and it was placed in an eurythermic group of species (Van den Hoek 1982). Tronholm et al. (2010b) reviewed the genus Dictyota for Europe using molecular data and a large dataset with sequences from several places of the world, and they restricted the occurrence of D. dichotoma to the northeastern Atlantic (Europe and Africa). The specimens from other oceans, previously identified as D. dichotoma, were assigned to other species and so this taxon should not be considered as a cosmopolitan species. So, the native range of D. dichotoma on the European Atlantic coasts was established, from Scotland and southern Norway to Portugal, the Mediterranean Sea and most of the Macaronesian islands, except for the tropical Cape Verde where it was not found (Van den Hoek 1982; Schnetter et al. 1987; Tronholm et al. 2010b). Despite the occurrence of D. dichotoma in South Africa there had been questioned by De Clerck (2003), Tronholm et al. (2010b) confirmed its presence using a psbA marker. This intriguing record could not be attributed to natural or introduced origin by these authors. On the Atlantic coast of South America there are many records of misidentification of species of Dictyota as D. dichotoma in earlier phycological studies (Dunal 1833; Martius et al. 1833; Saint-Hilaire 1833; Montagne 1839). Recently, D. dichotoma has been considered as a cryptogenic species in Argentina (Raffo et al. 2014; Schwindt et al. 2014; Gauna et al. 2015). Exotic species are one of the major threats to biodiversity and one of the major concerns throughout the oceans (Schwindt et al. 2014; Abreu et al. 2016). Invasion of marine macroalgae have been reported around the world in recent decades, such as Undaria pinnatifida (Harvey) Suringar, Sargassum muticum (Yendo) Fensholt, Codium fragile ssp. fragile (Suringar) Hariot, Caulerpa taxifolia (M. Vahl) C. Agardh and Schizymenia dubyi (Chauvin ex Duby) J. Agardh (Casas et al. 2004, 2008; Raffo et al. 2009; Irigoyen NEW ZEALAND JOURNAL OF BOTANY 295 et al. 2011; Ramírez et al. 2012). Recently, Schwindt et al. (2014) sampled six of the 10 main marine ports in Argentina and found the presence of 32 non-indigenous taxa, including exotic and cryptogenic taxa. This study aims to confirm the molecular identity of specimens previously identified as Dictyota dichotoma from Argentina and discuss the possible origin of the South Atlantic populations. Materials and methods Dictyota specimens were collected from Las Grutas, San Matías Gulf (40°48′ S, 64°48′ W) in 2014 from a population previously studied by Gauna et al. (2013, 2015) with respect to the morphology of the specimens, phenology and ecology (epiphytic communities). The algae were collected by SCUBA diving from the subtidal zone and then screened in the field to remove possible epiphytes. Each individual was separated into two subsamples, one of them was preserved in silica gel for molecular studies and the other was preserved in 4% formalin solution, and deposited at the Herbarium of Universidad Nacional del Sur (BBB; Bahia Blanca, Argentina). DNA was extracted using HiPurA Plant Genomic DNA Miniprep Purification Kit (MolBio HIMEDIA). The plastid-encoded PSII reaction centre D1 (psbA), NADH dehydrogenase subunit 1 (nad1) and Rubisco large subunit (rbcL) were amplified via polymerase chain reaction (PCR) utilising the primers from Tronholm et al. (2010b) and Bittner et al. (2008), and then sequenced by Macrogen Inc. Korea. The sequences were edited on Mega 6.0 (Tamura et al. 2013) and then analysed with others retrieved from GenBank (www.ncbi.nlm.nih.gov/genbank) (Table S1). Three datasets were generated with alignments of 695 bp for nad1, 775 bp for psbA and 1149 bp for rbcL. The phylogenetic reconstructions were performed for each gene separately. The concatenation would not be reliable in this case because sequences available on GenBank are from distinct vouchers and from different locations. The evolution model of GTR + I + G was obtained by jModelTest 2.14 for each marker (Darriba et al. 2012) and used in the Bayesian Inference (BI) analysis on MrBayes 3.1.2 (Ronquist et al. 2012). For nad1 and psbA, BI was carried out with 1 million generations in 2 runs and 4 chains, sampling every 1000th generation, discarding the first 30 and 50 trees for nad1 and psbA, respectively. For rbcL, BI was carried out with 2 million generations in 2 runs and 4 chains, sampling every 1000th generation, discarding the first 50 trees. The maximum likelihood (ML) analysis was performed on Mega 6.0, using the GTR + I + G model with bootstrap of 1000 replications. The gene with a higher number of Dictyota dichotoma sequences available from GenBank was psbA and therefore it was used to build a haplotype network with Dnasp v.5 (Librado & Rozas 2009) and Network v.5 by median joining. The initial alignment used the 220 available sequences (Table S2). Then, the shorter sequences and those with many missing data were removed from the analysis, leaving 149 sequences. To account for the disparity in the number of sequences available from different geographic regions/localities, up to 10 sequences/haplotypes per population were used in the analysis, with sequences retrieved from samples collected from up to 100 km apart considered as part of the same population. The final dataset used in the haplotype network analysis included 68 sequences with 638 bp (Table S2). 296 E. A. P. LOPES-FILHO ET AL. Results The topology of the trees for psbA, rbcL and nad1 obtained in the present study agreed with previous studies (Tronholm et al. 2010a, 2010b). The samples from Argentina formed a clade with genuine European sequences of D. dichotoma. For psbA the clade with sequences from Argentina and from the neotype elected by Tronholm et al. (2010b) obtained bootstrap values of 100% (ML) and 1.00 (BI) (Figure 1). For rbcL (Figure S1) high values of bootstrap and posterior probability were obtained for the clade with sequences from Argentina and Ireland. The same high values were observed for nad1 (Figure S2) for the clade with sequences from Argentina and France. The haplotype network (Figure 2) revealed seven haplotypes in the northeast Atlantic Ocean that are geographically divided into two groups: Atlantic–Mediterranean and Canarian. The Atlantic–Mediterranean group consisted of two haplotypes, with H1 being the most widespread and present on the Atlantic coasts of the Iberian Peninsula, Figure 1. Phylogenetic tree based on psbA sequences, presenting a consensus topology estimated by Maximum Likelihood (ML) and Bayesian Inference (BI) analyses. The numbers associated with each branch represent the statistical support values (only values above 95% are shown), where the first is the bootstrap values from ML and the second is the posterior probability from BI. NEW ZEALAND JOURNAL OF BOTANY 297 Figure 2. Haplotype diversity of psbA from Dictyota dichotoma. The red dots are the locations where the sequences of D. dichotoma on GenBank come from. a, Haplotype network of psbA showing the two groups: Atlantic–Mediterranean and Canarian; b, distribution of the seven haplotypes in the North Atlantic population; c, South Atlantic populations with the single haplotype H1. France, the British Islands and the North Sea, the entire Mediterranean Sea and parts of the Macaronesian islands (Azores and Madeira archipelagos). The second haplotype (H2) seems to be less frequent in general and was only detected in the Gulf of Lion (France), in the Mediterranean Sea. Samples from Argentina and South Africa corresponded to the H1 haplotype. The Canarian group consisted of five haplotypes (H3, H4, H5, H6 and H7) that are almost restricted to Macaronesia (Canary and Madeira islands), where H5 is the most common one, widely found throughout these islands. The H3 haplotype from the Canarian group is the only one that was detected away from these islands in the Gulf of Lion. Discussion The higher haplotype diversity observed for D. dichotoma in the northeastern Atlantic (Figure 2) is consistent with the hypothesis presented by Tronholm et al. (2010b) explaining the low diversity of the genus Dictyota in Europe. This hypothesis states that, following the desiccation of the Mediterranean Sea which caused the extinction of most of the marine biota during the Messinian salinity crisis (6.8–5.3 Ma), the Mediterranean Sea was recolonised by Atlantic species from adjacent areas, such as the Macaronesian islands and the northwestern African coast, when it was re-flooded (Coll et al. 2010; Tronholm et al. 2010b). The results from the present study demonstrate that European Atlantic and Macaronesian populations of D. dichotoma were probably separated during this vicariant event and, later, northern populations (haplotype H1) were successful in recolonising the Mediterranean Sea and subsequently reached Macaronesia (Azores and Madeira 298 E. A. P. LOPES-FILHO ET AL. archipelagos). Despite having a greater diversity, the Canarian group remained restricted to Macaronesia with the only exception being haplotype H3, in contrast to what seems to have happened with other Dictyota species from Macaronesia that were able to recolonise the Mediterranean Sea (Tronholm et al. 2010b). However, the presence of the H1 haplotype in the South Atlantic would contradict this scenario. The results of Tronholm et al. (2012), using the relaxed molecular clock for the genus, revealed D. dichotoma as the only extant species of one of the oldest lineages in the genus, which split early from the others c. 44.37 Ma (53–34 Ma), and being one of the first clades to disperse through the Tethys Seaway to the westernmost part of the Tethys realm where it most likely evolved and later dispersed to the Atlantic coast of Europe and Macaronesia. Since the opening of the South Atlantic (100–80 Ma) the northward paleocurrents along the African coast allowed the water transport of the Tethys Sea to the Atlantic via Southern Africa while most of the transport continued on account of the circum-equatorial circulation directly to the North Atlantic until 12–18 Ma (Stille 1992; Stille et al. 1996; Cowman & Bellwood 2013), when the circulation in the North Atlantic was governed by the Pacific– North Atlantic current (Iturralde-Vinent 2006). Therefore, the colonisation of the South Atlantic coast of Africa by D. dichotoma would have been prevented because it would have been against the direction of the ocean currents. Furthermore, there was no connection between the westernmost part of the Tethys realm and South America, thereby a warm paleocurrent (probably originating in the Caribbean) crossed the Atlantic coast of South America to Patagonia and Tierra del Fuego, where the marine communities were tropical until middle–late Miocene (Del Río 2004a, 2004b; Le Roux 2012). The hypotheses of trans-oceanic dispersal in this scenario fail mainly due to biology of D. dichotoma demonstrated by culture experiments in which this species does not possess affinities to tropical conditions (Biebl 1959; Bogaert et al. 2016) which would be required for it to be successfully dispersed along the South American coast up until the late Miocene. The adequate temperate marine conditions for D. dichotoma in the southwestern Atlantic were only established after the complete development of the Circumpolar Antarctic Current, which led to the full operation of the Malvinas/Falklands Current and the establishment of the Benguela Upwelling System in the middle–late Miocene (10–9 Ma). As a consequence, there was a decrease in the seawater temperature in Patagonia and along the southwestern African coast (Heinrich et al. 2011; Rommerskirchen et al. 2011; Le Roux 2012), the extinction of the Patagonian tropical marine communities and the retraction of the warm Brazilian current to the north of Argentina/Uruguay (Del Río 2004a, 2004b). Therefore, it is difficult to explain how D. dichotoma would have reached the coast of Argentina through natural dispersion. Those facts agree with the current data that do not show any exclusive haplotypes for the South Atlantic populations (Argentina and South Africa) of D. dichotoma, which would be expected in the case of an old dispersal from the northeastern Atlantic (over 10 Ma), or any haplotype shared with the Canarian group, which would be expected in the case of a recent dispersal (less than 6 Ma) and so, against the direction of the ocean currents. The occurrence of the H1 haplotype in the South Atlantic Ocean is inconsistent with the hypothesis of natural dispersal with subsequent genetic differentiation and supports the hypothesis of human-mediated introduction, where propagules of D. dichotoma settled successfully in Argentina and in South Africa because both areas belong to temperate provinces, similar to its native area in the northeastern Atlantic (Spalding et al. 2007). NEW ZEALAND JOURNAL OF BOTANY 299 The first evidence of the presence of D. dichotoma in Argentina based on chemosystematics, rather than just morphology, is presented by Palermo et al. (1994) who studied a population from Nuevo Gulf and identified three prenylated diterpenes expected to occur (and its precursors) in this species (Amico et al. 1976; Fattorusso et al. 1976; Faulkner et al. 1977; Siamopoulou et al. 2004; Vallim et al. 2005). The locations where D. dichotoma was collected by Palermo et al. (1994) and for this study are near to two of the six main marine ports studied by Schwindt et al. (2014). These authors demonstrated that the collection areas are in natural bays with anthropic influences (but with abiotic conditions adequate for D. dichotoma, such as salinity and surface water temperature) and have high maritime activities, which enable the high percentage of non-indigenous marine taxa found in them. Port areas provide artificial structures that favour the introduction of exotic fouling/ benthic species (recruitment, survival and dispersal) which are mainly transported by ballast water (Schwindt et al. 2014; Abreu et al. 2016; Lin & Zhan 2016). In South Africa, most of the introduced species reported in both the cool and warm temperate provinces are from the Northern Hemisphere (65%). At the Agulhas ecoregion, where the presence of D. dichotoma was confirmed (Tronholm et al. 2010b), 73 non-indigenous (alien, invasive or cryptogenic) taxa have been reported (Mead et al. 2011; Robinson 2015). Introduction of exotic species may be overlooked for decades (Abreu et al. 2016), especially when a group (such as Dictyota) has not been formally revised. It is not clear at present how D. dichotoma has affected the marine species in Argentina, especially on the northern parts of the Patagonian coast where the genus Dictyota (reported as D. dichotoma) is abundant (Casas et al. 2004; Gauna et al. 2015). For example, in the Gulf of San José and nearby areas (e.g. Nuevo Gulf and San Matias Gulf) D. dichotoma may cover up to 30% of the entire area at depths up to 10 m (Boraso de Zaixso & Zaixso 2007; A. Boraso, pers. comm., 2016). In the same way, the impact on native Dictyota species could not be estimated. Using the ecoregions proposed by Spalding et al. (2007), the known distribution of the genus Dictyota in Argentina (Figure S3) covers the final part of the Warm Temperate Southwestern Atlantic province and the Atlantic part of the Magallanic province. The persistent misidentification for almost two centuries (since Montagne 1839) resulted in D. dichotoma being considered as the only species in Argentina, and the reports of other species such as Canistrocarpus cervicornis (Kützing) De Paula & De Clerck (as Dictyota cervicornis Kützing), D. divaricata J.V. Lamouroux and D. dichotoma var. intricata (C.Agardh) Greville (Taylor 1939; Asensi 1966; Van den Hoek 1982; Boraso de Zaixso 1995; Mendoza & Nizovoy 2000; Piriz et al. 2003; Croce et al. 2015) were considered as representing a wide morphological variation of it (Boraso de Zaixso 2012). Although the genus has also been found in the Beagle Channel (A. Boraso, pers. comm., 2016), Mystikou et al. (2016) published the southernmost records of Dictyota in the southwestern Atlantic. Their molecular data confirmed that the species is distinct from D. dichotoma (as also verified in Figure 1), which is the first step towards uncovering the diversity of the genus in Argentina. The spread of D. dichotoma northward along the southwestern Atlantic from the Argentine populations is unexpected because of: 1. the Confluence Zone of the southward warm Brazilian current and northward cold Malvinas/Falklands current—which spans from about 25°S to 45°S—where the water masses are reflected eastward as a South Atlantic current, approximate average axis at 39°S (Bisbal 1995); and 2. the La Plata river (at 35°S), 300 E. A. P. LOPES-FILHO ET AL. which results in brackish and turbid water, as well as the lack of a suitable substrate that prevents the occurrence of seagrass and marine benthic macroalgae in the La Plata estuary region, and there is a depauperate flora on the Argentinian, Uruguayan and Brazilian coasts under its influence (Coll & Oliveira 1999; Calliari et al. 2003; Acha et al. 2008; Braga et al. 2008; Campos et al. 2008). Additionally, the only Uruguayan record of Dictyota (as D. dichotoma) was questioned by Coll & Oliveira (1999) because no specimens were found in any of the mentioned herbaria in the original study and no other specimens have been collected subsequently. Therefore, Dictyota species are separated by more than 1200 km between Mar del Plata (Argentina) and Torres (Brazil) due to the lack of hard substrate (Baptista 1977; Oliveira Filho 1977). In the southeastern Brazil ecoregion, which includes two important Brazilian harbours, there are seasonal coastal upwelling events and the cold (<18 °C) water masses can reside throughout the year in a deep benthic system (Coelho-Souza et al. 2012). These waters could sustain organisms with cold waters affinities, such as the brown alga Laminaria abyssalis Joly and Oliveira, which occurs below 50 m (Guimarães et al. 1986). Moreover, the intertidal Jolyna laminarioides Guimarães and Elachistiella leptonematoides Cassano, Yoneshigue-Valentin and Wynne (Valentin 2001; Cassano et al. 2004) only occur during the upwelling period. This can promote a temporary niche for the introduction of species with cold water affinities. There is no evidence, so far, of the presence of D. dichotoma and most of the morphologically similar specimens are D. menstrualis (Hoyt) Schnetter, Hörning and Weber-Peukert (unpubl. data). Recent reports of D. dichotoma and D. dichotoma var. intricata by Villaça et al. (2010) and Crespo et al. (2014) are due to the citation of old studies. In conclusion, the natural dispersal of the species from the northeast Atlantic to Argentina and South Africa would be unlikely, as this temperate species would have to cross the whole equatorial and tropical Atlantic Ocean against the direction of the currents. The occurrence of a single, and the most common, haplotype of the Atlantic–Mediterranean group in the South Atlantic populations, suggests that D. dichotoma was introduced. The human-assisted introduction of other taxa of Dictyotaceae has already been reported elsewhere, such as the cases of Rugulopteryx okamurae (E.Y. Dawson) I.K. Hwang, W.J. Lee and H.S. Kim (Verlaque et al. 2009), Dictyota cyanoloma Tronholm, De Clerck, A. Gómez-Garreta and Rull Lluc (Tronholm et al. 2010b; García et al. 2016) and Dictyota furcellata (C.Agardh) Greville (Nelson & Wilcox 2010). Future studies to include more comprehensive sampling are necessary and may reveal whether the introduction of D. dichotoma has occurred once or multiple times, its actual geographical range in Argentina and how it has affected the benthic marine community. Acknowledgements We would like to thank Dr Alicia Boraso for her valuable information and Clariana Ferraz Sampaio for proof reading the English version. Also, the two anonymous referees for suggestions that improved the manuscript. Associate Editor: Dr Roberta D’Archino. Disclosure statement No potential conflict of interest was reported by the authors. NEW ZEALAND JOURNAL OF BOTANY 301 Funding This work was supported by Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ E-26/111/397/2012) and by the scholarships received by EAPLF during his final year in college (UNIRIO) and during his Master’s Programme (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES). ORCID Joel Campos De Paula http://orcid.org/0000-0001-5852-7516 References Abreu NMN, Marçal I, Duarte AB, Pitombo FB, Vilasboa A, Gusmao J. 2016. Microsatellite markers for barnacle studies: isolation and characterization of polymorphic microsatellite loci from the invasive barnacle Megabalanus coccopoma (Darwin, 1854) and its cross-amplification in the Southern Atlantic endemic species Megabalanus vesiculosus (Darwin, 1854). Biochem Syst Ecol. 66:224–228. Acha EM, Mianzan H, Guerrero R, Carreto J, Giberto D, Montoya N, Carignan M. 2008. An overview of physical and ecological processes in the Rio de La Plata Estuary. Cont Shelf Res. 28:1579– 1588. Amico V, Oriente G, Piattelli M, Tringa C, Fattorusso E, Magno S, Mayol L. 1976. Dilphol, a new ten-membered-ring diterpene alcohol from the brown alga Dilophus ligulatus. JCS Chem Comm. 24:1024–1025. Asensi AO. 1966. Guía para reconocer los géneros de algas pardas de la Argentina [Guide to recognise the brown algae genera from Argentina]. Contrib Inst Antárt Argent. 103:1–51. Spanish. Baptista LRM. 1977. Flora ilustrada do Rio Grande do Sul. Flora marinha de Torres [Illustrated flora from Rio Grande do Sul. Marine flora from Torres]. Bolm Inst Biociênc (Bot.). 37:1–246. Portuguese. Biebl R. 1959. Temperatur-und osmotische Resistenz von Meeresalgen der bretonischen Küste [Temperature and osmotic resistance of the marine algae from Breton coast]. Protoplasma. 50:217-242. German Bisbal GA. 1995. The Southeast South American shelf large marine ecosystem. Evolution and components. Mar Policy. 19:21–38. Bittner L, Payri CE, Couloux A, Cruaud C, De Reviers B, Rousseau F. 2008. Molecular phylogeny of the Dictyotales and their position within the Phaeophyceae, based on nuclear, plastid and mitochondrial DNA sequence data. Mol Phylog and Evol. 49:211–226. Bogaert K, Beeckman T, De Clerck O. 2016. Abiotic regulation of growth and fertility in the sporophyte of Dictyota dichotoma (Hudson) J.V. Lamouroux (Dictyotales, Phaeophyceae). J Appl Phycol. 28:2915–2924. Boraso de Zaixso A. 1995. Algas bentónicas de Puerto Deseado (Santa Cruz). Composición de la flora luego de la erupción del volcan Hudson [Benthic algae from Puerto Deseado (Santa Cruz). Flora composition after the eruption of Hudson Volcano]. Nat Patagon Ser Cienc Biol. 3:129–152. Spanish. Boraso de Zaixso AL. 2012. Elementos para el estudio de las macroalgas marinas de Argentina [Elements to the study of the marine macroalgae from Argentina]. Instituto de Desarrollo Costero (IDC). Comodoro Rivadavia: Editorial Universitaria de la Patagonia. Boraso de Zaixso AL, Zaixso JM. 2007. Algas marinas bentónicas [Benthic marine algae]. In: Boltovskoy D, editor. Atlas de Sensibilidad Ambiental de la costa y el Mar Argentino [Internet]. República Argentina: Secretaria de Ambiente y Desarrollo Sustentable. Available from: http://atlas.ambiente.gov.ar 302 E. A. P. LOPES-FILHO ET AL. Braga ES, Chiozzini VC, Berbel GBB, Maluf JCC, Aguiar VMC, Charo M, Molina D, Romero SI, Eichler BB. 2008. Nutrient distributions over the Southwestern South Atlantic continental shelf from Mar del Plata (Argentina) to Itajaí (Brazil): winter–summer aspects. Cont Shelf Res. 28:1649–1661. Calliari D, Defeo O, Cervetto G, Gómez M, Giménez L, Scarabino F, Brazeiro A, Norbis W. 2003. Marine life of Uruguay: critical update and priorities for future research. Gayana. 67:341–370. Campos EJD, Piola AR, Matano RP, Miller JL. 2008. PLATA: a synoptic characterization of the southwest Atlantic shelf under influence of the Plata River and Patos Lagoon outflows. Cont Shelf Res. 28:1551–1555. Casas G, Scrosati R, Piriz ML. 2004. The invasive kelp Undaria pinnatifida (Phaeophyceae, Laminariales) reduces native seaweed diversity in Nuevo Gulf (Patagonia, Argentina). Biol Invasions. 6:411–416. Casas GN, Piriz ML, Parodi ER. 2008. Population features of the invasive kelp Undaria pinnatifida (Phaeophyceae: Laminariales) in Nuevo Gulf (Patagonia, Argentina). J Mar Biol Assoc UK. 88:11–28. Cassano V, Yoneshigue-Valentin Y, Wynne MJ. 2004. Elachistiella leptonematoides gen. et sp. nov. (Elachistaceae, Phaeophyceae) from Brazil. Phycologia. 43:329–340. Coelho-Souza SA, López MS, Guimarães JRD, Coutinho R, Candella RN. 2012. Biophysical interactions in the Cabo Frio upwelling system, southeastern Brazil. Braz J Oceanogr. 60:353–365. Coll J, Oliveira EC. 1999. The benthic marine algae of Uruguay. Bot. Mar. 42:129–135. Coll M, Piroddi C, Steenbeek J, Kaschner K, Ben Rais Lasram F, Aguzzi J, Ballesteros E, Bianchi CN, Corbera J, Dailianis T, et al. 2010. The biodiversity of the Mediterranean Sea: estimates, patterns, and threats. PLoS ONE. 5:e11842. doi:10.1371/journal.pone.0011842 Cowman PF, Bellwood DR. 2013. Vicariance across major marine biogeographic barriers: temporal concordance and the relative intensity of hard versus soft barriers. Proc R Soc B. 280:20131541. doi:10.1098/rspb.2013.1541 Crespo TM, Bahia RG, Maneveldt GW, Amado Filho GM. 2014. Floristic composition of crustose coralline algae from the St. Peter and St. Paul Archipelago, a summit of the Mid-Atlantic Ridge. Phytotaxa. 190:17–37. Croce ME, Gauna MC, Fernández C, Parodi ER. 2015. Intertidal seaweeds from North Atlantic Patagonian coasts, Argentina. Check List. 11:1739. doi:10.15560/11.5.1739 Darriba D, Taboada G, Doallo R, Posada D. 2012. Jmodeltest 2: more models, new heuristics and parallel computing. Nat Methods. 9:772. doi:10.1038/nmeth.2109 De Clerck O. 2003. The genus Dictyota in the Indian Ocean. Opera Botánica Belgica 13. Meise: National Botanic Garden of Belgium. De Clerck O, Leliaert F, Verbruggen H, Lane CE, De Paula JC, Payo DA, Coppejans E. 2006. A revised classification of the Dictyoteae (Dictyotales, Phaeophyceae) based on Rbcl and 26s ribosomal Dna sequence analyses. J Phycol. 42:1271–1288. Del Río CJ. 2004a. Relaciones biogeográficas entre los moluscos del Mioceno tardío y reciente del Atlántico Sudoccidental [Biogeographic relations between late Miocene and recent molluscs from the Southwestern Atlantic]. In: Aceñolaza FG, editor. Temas de la Biodiversidad del Litoral fluvial Argentino. Misceláneas 12. Tucumán: Instituto Superior de Correlación Geológica; p. 39–44. Del Río CJ. 2004b. Tertiary marine molluscan assemblages of eastern Patagonia (Argentina): a biostratigraphic analysis. J. Paleont. 78:1097–1122. De Paula JC, Vallim MA, Teixeira VL. 2011. What are and where are the bioactive terpenoids metabolites from Dictyotaceae (Phaeophyceae). Braz J Pharmacog. 21:216–228. Dunal F. 1833. Bulletin Bibliographique. “Voyage dans le District des Diamans et sur le litoral du Brésil” par M. Auguste de Saint-Hillaire, membre de l’Académie des sciences, etc [“Voyage to the Diamond District and to the southern coast from Brazil” by M. Auguste de Saint-Hillaire, member of the Academy of Sciences etc]. 2 vol. In-8° - Paris, Gide, 1833. Arch Bot. 2:444– 456. French. Fattorusso E, Magno S, Mayol L, Santacroce C, Sica D, Amico V, Oriente G, Piattelli M, Tringali C. 1976. Dictyol A and B, two novel diterpene alcohols from the brown alga Dictyota dichotoma. J Chem Soc, Chem Commun. 14:575–576. NEW ZEALAND JOURNAL OF BOTANY 303 Faulkner DJ, Ravi BN, Finer J, Clardy J. 1977. Diterpenes from Dictyota Dichotoma. Phytochemistry. 16:991–993. García M, Weitzmann B, Pinedo S, Cebrian E, Ballesteros E. 2016. First report on the distribution and impact of marine alien species in Coastal Benthic assemblages along the Catalan Coast. In: Munné A, Ginebreda A, Prat N, editor. Experiences from ground, coastal and transitional water quality monitoring. The EU water framework directive implementation in the catalan river basin district (Part II). The handbook of environmental chemistry 43. Cham, CH: Springer; p. 249–270. Gauna MC, Cáceres EJ, Parodi ER. 2013. Temporal variations of vegetative features, sex ratios and reproductive phenology in a Dictyota dichotoma (Dictyotales, Phaeophyceae) population of Argentina. Helgoland Mar Res. 67:721–732. Gauna MC, Cáceres EJ, Parodi ER. 2015. Spatial and temporal variability in algal epiphytes on Patagonian Dictyota dichotoma (Dictyotales, Phaeophyceae). Aquat Bot. 120:338–345. Genzano GN, Rodriguez GM. 1998. Associations between hydroid species and their substrates from the intertidal zone of Mar del Plata (Argentine). Misc Zool. 21:21–29. Guimarães SMPB, Braga MRA, Cordeiro-Marino M, Pedrini AG. 1986. Morphology and taxonomy of Jolyna laminarioides, a new member of the Scytosiphonales (Phaeophyceae) from Brazil. Phycologia. 25:99–108. Heinrich S, Zonneveld KAF, Bickert T, Willems H. 2011. The Benguela upwelling related to the Miocene cooling events and the development of the Antarctic Circumpolar Current: evidence from calcareous dinoflagellate cysts. Paleoceanography. 26:PA3209. doi:10.1029/2010PA002065 Hwang IK, Kim HS, Lee WJ. 2005. Polymorphism in the brown alga Dictyota dichotoma (Dictyotales, Phaeophyceae) from Korea. Mar Biol. 147:999–1015. Irigoyen AJ, Trobbiani G, Sgarlatta MP, Raffo MP. 2011. Effects of the alien algae Undaria pinnatifida (Phaeophyceae, Laminariales) on the diversity and abundance of benthic macrofauna in Golfo Nuevo (Patagonia, Argentina): potential implications for local food webs. Biol Invasions. 13:1521–1532. Iturralde-Vinent MA. 2006. Meso-Cenozoic Caribbean paleogeography: implications for the historical biogeography of the region. Int Geol Rev. 48:791–827. Lamouroux JVF. 1809. Observations sur la physiologie des algues marines, et description de cinq noveaux genres de cette famille [Observations about the physiology of the marine algae and the description of five new genera from this family]. Nouv Bull Sci Soc Philom, Paris. 1:330– 333. French. Le Roux JP. 2012. A review of Tertiary climate changes in southern South America and the Antarctic Peninsula. Part 1: Oceanic conditions. Sediment Geol. 247–248:1–20. Librado P, Rozas J. 2009. DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics. 25:1451–1452. Lin Y, Zhan A. 2016. Population genetic structure and identification of loci under selection in the invasive tunicate, Botryllus schlosseri, using newly developed EST-SSRs. Biochem Syst Ecol. 66:331–336. Littler DS, Littler MM. 2000. Caribbean reef plants: an identification guide to the reef plants of the Caribbean, Bahamas, Florida and Gulf of Mexico. Washington, DC: OffShore Graphics, Inc. Martius KFP, Eschweiler GG, Esenbeck CGDN. 1833. Flora Brasiliensis; seu Enumeratio plantarum in Brasilia tam sua sponte quam accedente cultura provenientium, quas in itinere auspiciis Maximiliani Josephi I. Bavariae regis annis 1817-1820 peracto collegit, partim descripsit; alias a Maximiliano seren. Principe Widensi, sellovio aliisque advectas addidit [Brazilian Flora: Enumeration of the plants from Brazil of natural and cultivated origins, under the care of Maximilian Joseph I, king of Bavaria, collected during the years 1817-1820 etc]. 1. 1: IV + 390 (Algae 1-50), 8 vol. Stuttgartiae et Tubingen: Sumptibus J.G. Cottae, p. 1–50. Mead A, Carlton JT, Griffiths CL, Rius M. 2011. Revealing the scale of marine bioinvasions in developing regions: a South African re-assessment. Biol Invasions. 13:1991–2008. Mendoza ML, Nizovoy A. 2000. Géneros de macroalgas marinas de la Argentina fundamentalmente de Tierra del Fuego [Marine macroalgae genera from Argentina, especially from Tierra 304 E. A. P. LOPES-FILHO ET AL. del Fuego]. Secretaria de Desarrollo y Planeamiento de la Provincia de Tierra del Fuego. Ushuaia: Tierra del Fuego. Montagne C. 1839. Cryptogames de la Patagonie [Cryptogamic Plants from Patagonia]. In: Voyage Dans L’amérique Méridionale (Le Brésil, La République Orientale De L’uruguay, La République Argentine, La Patagonie, La République Du Chili, La République de Bolivia, La République Du Pérou), Exécité Pendant Les Années 1826, 1827, 1828, 1829, 1830, 1831, 1832 Et 1833. Vol. 7: Botanique. Part I. Sertum Patagonium. (D’orbigny, A. Eds), p. 1–19. Paris. Moore PG, Eastman LB. 2015. The tube-dwelling lifestyle in crustaceans and its relation to feeding. In: Thiel M, Watling L, editors. Lifestyles and feeding biology. The natural history of the crustacea.V.2. New Tork: Oxford University Press; p. 35–77. Mystikou A, Asensi AO, De Clerck O, Müller DG, Peters AF, Tsiamis K, Fletcher KI, Westermeier R, Brickle P, West P, Küpper FC. 2016. New records and observations of macroalgae and associated pathogens from the Falkland Islands, Patagonia and Tierra del Fuego. Bot Mar. 59:105–121. Nelson WA, Wilcox MD. 2010. Rosenvingea (Ectocarpales, Scytosiphonaceae) – a new brown macroalgal record for New Zealand. New Zeal J Bot. 48:193–196. Oliveira Filho EC. 1977. Algas marinhas bentônicas do Brasil [Benthic marine algae from Brazil] [Thesis]. São Paulo (SP): Universidade de São Paulo. Palermo JA, Bernardo JJ, Seldes AM. 1994. Dictyol-D-2b-acetate and other diterpenoids from the brown alga Dictyota dichotoma. Ann Assoc Quím Argent. 82:355–358. Piriz ML, Eyras MC, Rotagno CM. 2003. Changes in biomass and botanical composition of beach coast seaweeds in a disturbed coastal area from Argentine Patagonia. J Appl Phycol. 15:67–74. Raffo MP, Eyras MC, Iribarne OO. 2009. The invasion of Undaria pinnatifida to a Macrocystis pyrifera kelp in Patagonia (Argentina, south-west Atlantic). J Mar Biol Assoc UK. 89:1571–1580. Raffo MP, Lo Russo V, Schwindt E. 2014. Introduced and native species on rocky shore macroalgal assemblages: zonation patterns, composition and diversity. Aquat Bot. 112:57–65. Ramírez ME, Nuñez JD, Ocampo EH, Matula CV, Suzuki M, Hashimoto T, Cledón M. 2012. Schizymenia dubyi (Rhodophyta, Schizymeniaceae), a new introduced species in Argentina. New Zeal J Bot. 50:51–58. Robinson T. 2015. Marine invasions in South Africa: patterns and trends. Quest. 11:44–45. Rommerskirchen F, Condon T, Mollenhauer G, Dupont L, Schefuss E. 2011. Miocene to Pliocene development of surface and subsurface temperatures in the Benguela Current system. Paleoceanography. 26:PA3216. doi:10.1029/2010PA002074 Ronquist F, Teslenko M, Der Mark PV, Ayres DL, Darling A, Hohna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP. 2012. MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst Biol. 61:539–542. Saint-Hilaire A. 1833. Voyage dans le District des Diamans et sur le litoral du Brésil [Voyage to the Diamond District and to the southern coast from Brazil]. Vol.1, i-xx+402pp. Paris. Schnetter R, Hörnig I, Weber-Peukert G. 1987. Taxonomy of some North Atlantic Dictyota species (Phaeophyta). Hydrobiologia. 151-152:193–197. Schwindt E, Gappa JL, Raffo MP, Tatian M, Bortolus A, Orensanz JM, Alonso G, Diez ME, Doti B, Genzano G, et al. 2014. Marine fouling invasions in ports of Patagonia (Argentina) with implications for legislation and monitoring programs. Mar Environ Res. 99:60–68. Siamopoulou P, Bimplakis A, Iliopoulou D, Vagias C, Cos P, Berghe DV, Roussis V. 2004. Diterpenes from the brown algae Dictyota dichotoma and Dictyota linearis. Phytochemistry. 65:2025–2030. Spalding MD, Fox HE, Allen GR, Davidson N, Ferdaña ZA, Finlayson M, Halpern BS, Jorge MA, Lombana A, Lourie SA, et al. 2007. Marine ecoregions of the world: a bioregionalization of coastal and shelf areas. BioScience. 57:573–583. Stachowicz JJ, Hay ME. 2000. Geographic variation in camouflage specialization by a decorator crab. Am Nat. 156:59–71. Stille P. 1992. Nd-Sr isotope evidence for dramatic changes of paleocurrents in the Atlantic Ocean during the past 80 m.y. Geology. 20:387–390. Stille P, Steinmann M, Riggs SR. 1996. Nd isotope evidence for the evolution of the paleocurrents in the Atlantic and Tethys Oceans during the past 180 Ma. Earth Planet Sc Lett. 144:9–19. NEW ZEALAND JOURNAL OF BOTANY 305 Széchy MTM, De Paula JC. 2016. Phaeophyceae: Lista de Espécies da Flora do Brasil [Phaeophyceae: Checklist of the Brazilian flora] [Internet]. Rio de Janeiro: Jardim Botânico do Rio de Janeiro. [cited 2016 Sept 25]. Available from: http://floradobrasil.jbrj.gov.br/jabot/ floradobrasil/FB99411. Portuguese. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. Mega6: molecular evolutionary genetics analysis version 6.0. Mol Phylogenet Evol. 30:2725–2729. Taylor WR. 1939. Algae collected by the “Hassler”, “Albatross” and Schmitt expeditions. II. Marine algae from Uruguay, Argentina, the Falkland Islands, and the Strait of Magellan. Papers Mich Ac Sc Arts Let. 24:127–164. Tronholm A, Leliaert F, Sansón M, Afonso-Carrillo J, Tyberghein L, Verbruggen H, De Clerck O, Crandall KA. 2012. Contrasting geographical distributions as a result of thermal tolerance and long-distance dispersal in two allegedly widespread tropical brown algae. Plos One. 7:e30813. doi:10.1371/journal.pone.0030813 Tronholm A, Sansón M, Afonso-Carrillo J, De Clerck O. 2008. Distinctive morphological features, life-cycle phases and seasonal variations in subtropical populations of Dictyota dichotoma (Dictyotales, Phaeophyceae). Bot Mar. 51:132–144. Tronholm A, Sansón M, Afonso-Carrillo J, Verbruggen H, De Clerck O. 2010a. Niche partitioning and the coexistence of two cryptic Dictyota (Dictyotales, Phaeophyceae) species from the Canary Islands. J Phycol. 46:1075–1087. Tronholm A, Steen F, Tyberghein L, Leliaert F, Verbruggen H, Siguan MAR, De Clerck O. 2010b. Species delimitation, taxonomy and biogeography of Dictyota in Europe (Dictyotales, Phaeophyceae). J Phycol. 46:1301–1321. Valentin JL. 2001. The Cabo Frio upwelling system, Brazil. In: Seeliger U, Kjerfve B, editors. Coastal Marine Ecosystems of Latin America. Ecol Stud. 144. Berlin: Springer; p. 97–105. Vallim MA, De Paula JC, Pereira RC, Teixeira VL. 2005. The diterpenes from Dictyotacean marine brown algae in the Tropical Atlantic American region. Biochem Syst Ecol. 33:1–16. Van den Hoek C. 1982. Phytogeographic distribution groups of benthic marine algae in the north Atlantic Ocean. A review of experimental evidence from life history studies. Helgol Meeresunters. 35:153–214. Verlaque M, Steen F, De Clerck O. 2009. Rugulopteryx (Dictyotales, Phaeophyceae), a genus recently introduced to the Mediterranean. Phycologia. 48:536–542. Villaça R, Fonseca AC, Jensen VK, Knoppers B. 2010. Species composition and distribution of macroalgae on Atol das Rocas, Brazil, SW Atlantic. Bot Mar. 53:113–122.