researcharticle - hindawi publishing...

Research ArticleThe Ulva spp Conundrum What Does the Ecophysiology ofSouthern Atlantic Specimens Tell Us

Vin-cius Peruzzi Oliveira 1 Barbara Lage Ignacio2 Nuno Tavares Martins3

Leticia Dobler4 and Alex Enrich-Prast56

1Universidade Federal do Rio de Janeiro (UFRJ) Departamento de Biologia Marinha Brazil2Universidade Federal de Sao Paulo (UNIFESP) Departamento de Ciencias do Mar Brazil3Universidade de Sao Paulo (USP) Instituto de Biociencias Brazil4Universidade Federal do Rio de Janeiro (UFRJ) Instituto de Quımica Brazil5Universidade Federal do Rio de Janeiro (UFRJ) Departamento de Botanica Brazil6Linkoping University Department of Thematic Studies Sweden

Correspondence should be addressed to Vinıcius Peruzzi Oliveira viniciusperuzzigmailcom

Received 20 September 2018 Revised 5 December 2018 Accepted 7 February 2019 Published 3 March 2019

Academic Editor Shamsul Hayat

Copyright copy 2019 Vinıcius PeruzziOliveira et alThis is an open access article distributed under theCreative Commons AttributionLicensewhichpermits unrestricteduse distribution and reproduction in anymedium provided the original work is properly cited

Species of the genus Ulva are common in anthropogenically disturbed areas and have been reported as the cause of green tides inmany areas of the world In addition they rank among the main marine groups used in a wide range of commercial applicationsBy displaying few distinctive morphological characters some taxonomical identifications are difficult and the genus is under aconundrum Our aims were to provide ecophysiological information about three Ulva species in response to abiotic factors and toevaluate the proposal of ecophysiological information and the chlorophyll-a fluorescence technique as auxiliary tool to resolve thelong-standing taxonomic confusion We hypothesize that three cooccurring specimens (U fasciataDelileU lactuca Linnaeus andU rigida C Agardh) have different ecophysiological responses (as measured by the effective quantum yield of photosystem II bypulse amplitude modulated fluorometers) under manipulated conditions of temperature and nutrient concentration Ulva lactucaandU rigida showed different photosynthetic efficiencies related to temperaturewhereas no differencewas recorded forU fasciataindividuals These results provide a reasonable explanation for the variability in spatial and temporal abundance of these speciesof Ulva on rocky shores We proposed the use of ecophysiological information by chlorophyll-a fluorescence as an auxiliary toolto corroborate the taxonomic distinction of Ulva species We reinforce the statement of U fasciata and U lactuca as distinct validspecies

1 Introduction

The genus Ulva (Chlorophyta) comprises cosmopolitanmacroalgae that inhabit a gradient from freshwater to fullysaline shallow environments [1] Species of this genus rankamong the first species to be established in disturbed environ-ments because of their morpho-physiological features thatallow tolerance to wide range of environmental conditions[2ndash4] Some species of Ulva have been emphasized due toreports as the cause of green tides in many parts of the world[5] Additionally numerous studies have also demonstratedfunctions in bioremediation of eutrophic environments and

uses of their biomass and carbohydrate content in thepharmaceutical food and energy chain [6ndash8]

Ongoing advances in molecular biology have driven tax-onomic revisions and morphological features have lost cred-ibility as the primary species delimitation criterion for manygroups (see [9] for algae) Ulva is one of the genera involvedin a taxonomic conundrum that entangles morphological ormolecular data for species identification mainly involvingdistinction around U lactuca and U fasciata (see [1 10ndash13])In spite of the undeniable contribution of molecular methodsto taxonomy (see [14ndash17]) there is no easy way to determinethe accurate match between the GenBank sequences and the

HindawiJournal of Marine BiologyVolume 2019 Article ID 5653464 6 pageshttpsdoiorg10115520195653464

2 Journal of Marine Biology

morphological characterization of most studies which hasled to uncertainty and sometimes to misinterpretation (see[1 11])

In the latter context physiological markers that areassociated with the photosynthesis process may be espe-cially helpful Photosynthetic organisms adjust the opera-tion of their photosynthetic apparatus to optimize or pre-serve metabolism under stress which can be detected bychlorophyll-a fluorescence variation [18 19] In this casethe evaluation of the effective quantum yield of PSII bypulse amplitude modulated (PAM) fluorometers has beenproposed as an attractive measure of the ecophysiologicalcondition [20ndash24] This technique is notable for performinga nonintrusive real-time analysis under both laboratory andfield conditions [25] and has been applied for establishing thedifferences among morphotypes of macroalgae [26]

Under this background the aim of this study was to pro-vide comparative ecophysiological information about threecooccurring species of Ulva in the Southwestern Atlantic(Ulva rigida C Agardh Ulva lactuca Linnaeus and Ulvafasciata Delile) that were morphologically identified Weassessed the effect of temperature and nutrient variation onthe photosynthetic efficiency (by chlorophyll-a fluorescencemeasurements) of the three species The general testedhypothesis was that species would have different responses toboth of the tested factors We subsequently expect to provideuseful information for using PAM fluorometry of Ulva asa reliable tool for ecophysiological assessment and for thetaxonomic debate about Ulva

2 Material and Methods

21 Sampling The three species of Ulva (U fasciata U lac-tuca and U rigida) were collected during summer (January2010) in the intertidal zone at Prainha Beach (22∘ 571015840S 042∘011015840W) Rio de Janeiro Brazil Immediately the epibionts wereremoved and the macroalgae were kept in cool boxes withlocal seawater (sim30 120583M NO

3 3 120583M PO

4 19∘C and 35 PSU)

filtered through 10-120583mmesh filters

22 The Species The morphological taxonomy of the threestudied species (U fasciata U lactuca and U rigida) wasbased on botanical descriptions and inventories that consid-ered in-depth confrontation among the species descriptionsand the Southwestern Atlantic specimens [27ndash30] Individ-uals were prepared as herbarium vouchers and depositedin the herbarium of the Universidade Federal do Rio deJaneiroUFRJ (under the accession numbers RFA 4244142442 42443)

23 Experimental Design All the individuals were main-tained in Erlenmeyer flasks in a proportion of 1 g of freshmass Lminus1 pasteurized oligotrophic ocean water with addedtrace nutrients (lt1 120583M NH

4 lt1 120583M NO

3 lt2 120583M NO

2 lt1

120583MPO4) at 20∘C and 200 120583mol photons mminus2 sminus1 (PAR pho-

tosynthetically active radiation) for 15 days This phase wasestablished to set all the macroalgae to the same nutritionaland physiological state before the manipulative experiment

This PAR was chosen because it is an intermediate value inrelation to the saturation parameter that is found in the P-Icurves (100ndash400 photons mminus2 sminus1)

After the nutrient deprivation period six treatments wereapplied using combinations of two temperatures and threeconcentrations of nutrients with 5 replicates per treatmentThe experiment per se lasted 10 hours The factor ldquoTemper-aturerdquo was represented by 20∘C and 30∘C and the factorldquoNutrientrdquo was represented by different additions of NaNO

3

minus

and Na2HPO412H2O of Von Stoschrsquos enriched seawater

(VSE 500 120583MNO3

minus and 30 120583MPO4

minus) [31]The experimentalsetup values of temperatures are related to the temperatureat the upper intertidal fringe at the sampling site whichranges between 18∘C (high tide) and 32∘C (low tide) [29]and the nutrient concentrations were set as 01 VSE 02 VSEand 05 VSE These concentrations refer to a gradient ofnutrient concentrations that were recorded in SouthwesternAtlantic [32 33] The treatment set as 01 VSE refers tonatural eutrophicated waters (upwelling events at collectingsite Arraial do Cabo Rio de Janeiro) [32] whereas 02 VSEand 05 VSE refer to the range of nutrient concentrations thatwere recorded in the human-disturbed Guanabara Bay Riode Janeiro [33]

24 Fluorescence Measurements The photosynthetic effi-ciency of the three species of Ulva in every combinationof temperature and nutrient concentration was charac-terized by chlorophyll-a fluorescence measurements usinga submersible diving-PAM fluorometer (Walz EffeltrichGermany) The measurements consisted of recording theeffective quantum yield of PSII which was calculated as Y= (F1015840m minus Ft)F

1015840

m where F1015840m is the maximum fluorescencein the light (obtained by a saturating actinic light pulse ndash8900 120583mol photons mminus2 sminus1 08 s) and Ft is the steady-state of fluorescence in the light [34 35] Considering thepossibility of physiological differences along the thallus [36]the fluorescence was measured in a similar region of thethallus in each individual

25 Data Analyses To determine the effects if any of tem-perature and nutrients on these three species of Ulva basedon the Y measurements a four-way analysis of variance wasperformedThe analysis included the (i) species (orthogonalfixed three levels) (ii) nutrients (orthogonal fixed three lev-els) (iii) temperature (orthogonal fixed two levels) and (iv)plot Erlenmeyer flasks (random and nested in the interactionof factors 1 and 2 two levels) The SNK post hoc test was usedto examine the nature of the differences that were detectedwith ANOVA (005 significance level) Statistical analyseswere performed using GMAV5 for Windows

3 Results

Differences in the Y were found for the interaction ofspecies and temperature (Table 1) For both U lactuca and Urigida the individuals that were incubated at 30∘C showed ahigher photosynthetic efficiency than did those incubated at20∘C whereas no difference in photosynthetic efficiency was

Journal of Marine Biology 3

Table 1 Four-way analyses of variance of the photosynthetic capacities (based on the effective quantum yield of the PSII measurements)of three species of Ulva (orthogonal fixed) that were incubated under different nutrient (orthogonal fixed three levels) and temperature(orthogonal fixed two levels) conditions Plot Erlenmeyer flasks where individuals were incubated (random and nested in the interactionof factors 1 and 2 two levels) SS is sums of squares DF is degrees freedom MS is mean of square and P-value lt005

Source SS DF MS F PSpecies (Sp) 00003 2 00001 018 084Temperature (Te) 00129 1 00129 940 002Nutrient (Nu) 00018 2 00009 064 056Plot (TetimesNu) 00082 6 00014 229 004lowast

SptimesTe 00072 2 00036 467 003lowast

SptimesNu 00082 4 00020 264 009SptimesPlot (TetimesNu) 00093 12 00008 129 023TetimesNu 00032 2 00016 115 038SptimesTetimesNu 00081 4 00020 263 009residual 00861 144 00006total 01452 179

080

079

078

077

076

075

Y

20∘C 30

∘C 20∘C 30

∘C 20∘C 30

∘CU fasciata U lactuca U rigida

Figure 1 Photosynthetic capacities of the three species of Ulvaunder two temperature conditions (based on the effective quan-tum yield of PSII measurements) Data from different nutrientconditions and plots (Erlenmeyer flasks where individuals wereincubated) were pooled n=30 Standard deviation values are notshown because of scale-bar limitation (left to right 0021 00230032 0019 0026 and 0035)

recorded for U fasciata individuals (Figure 1) Even when anANOVA was performed for only U fasciata no difference inphotosynthetic efficiency was found

4 Discussion

There are currently 403 species of Ulva in the literature ofwhich 131 have been flagged as accepted [37] Even withthe development of new tools for taxonomic precision theidentification in the practicemay differ fromauthor to authorIn this sense fast and real-time physiological assessmentcan be helpful to distinguish species and morphotypes ofmacroalgae

In this study two of the three species of Ulva (U lactucaand U rigida) showed differences in the Y as related to

temperature Although these species present large morpho-logical distinctions both showed a higher photosyntheticefficiency at the highest experimental temperature (30∘C)denoting the sensitivity of these species to this temperaturevalue The physiological differences can be linked to thehydrodynamic environment once they are under submer-gence for long periods of time In this case this effect oftemperature on photosynthetic metabolism is well known formacroalgae [38 39] and several positive effects on chemicalreactions andmolecular structures have been reported due tothe changes in protein compounds and enzymatic activities[40 41]

In contrast Ulva fasciata did not show differences inthe photosynthetic efficiency related to the temperaturevariation Specifically this is the most abundant of the threespecies in the sampling place dominant at the upper limitof the intertidal zone as also reported by Guimaraes andCoutinho [42] Its photosynthetic apparatus tolerates changesof at least 10∘C with no loss of efficiency as corroboratedby our study which can confer an ecological advantage andseems to support its relative spatial and temporal dominanceon rocky shores [38 43]

Despite their unequal abundances the three speciesare usually recorded throughout the year although thiscooccurrence is most evident during the summer [29] Thesummer period is characterized by high irradiance (ie hightemperatures on rocky shores) cold waters and a high inputof nutrients from the upwelling and rainfall runoff [44 45]Taken together these results suggest that the ecophysiologicaldifferences that were pointed in this study provide a reason-able explanation for the variability in the spatial and temporalabundance of these species of Ulva on rocky shores

It is important to highlight that despite the absenceof nutrient effects on the photosynthetic response of Ulvaspecies in our study we are not fully disregarding theimportance of this factor as a driver of the spatial andtemporal variability ofUlva species on rocky shores Nutrientenrichment favors ephemeral foliose macroalgae (see [3])by increasing the photosynthetic response [46] In fact the


genus Ulva is recognized by the affinity for nitrogen [47ndash49] and differences in photosynthetic efficiency have beenpredicted (if not among species at least among nutrient con-centrations) in contrast to the present results Consideringthe proper experimental design and applied manipulativemeasurements we attribute these findings to a saturated rateof nutrient availability light supply in our experiment

One of the most vexing taxonomic concerns refers to theU fasciata and U lactuca species which cooccur with Urigida Regarding U fasciata and U lactuca there is a long-standing taxonomic debate (see [1 10ndash13 50 51])Meanwhileseveral studies confirm U rigida as a consistent valid species[52ndash54] We corroborate the previous claim of Hiraokaet al [10] Shimada et al [50] and Kirkendale et al [1]Our study considered ecophysiological and morphological(based on [28ndash30]) data for specimens from SouthwesternAtlantic Therefore we propose the use of ecophysiologyas a complementary tool for taxonomic distinction amongmacroalgae based on chlorophyll-a fluorescence to enlightenthis debate with additional taxonomic evidence Similarmorphophysiological studies based on classic morphometryand photosynthetic responses (maximum electron transportrates ETRmax) suggested different species of Durvillaea(Phaeophyceae) for two morphotypes cooccurring in south-ern Chile [26]

However how this ecophysiology method would respondon other marine macroalgal taxa remains to be tested Weunderstand that it is hard to apply the methodology toidentification of freshly collected specimens or species in thefield without further investigation either by morphology ormolecularmethodsWe also recognize the undeniable contri-bution of morphology and molecular methods to taxonomyand our proposal aims to enlighten the debate with additionalevidence

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

The authors declare that they have no conflicts of interest

Acknowledgments

We are in debt to Mariana Mayer Pinto for her sup-port with the GMAV5 program We also thank the Con-selho Nacional de Desenvolvimento Cientıfico e Tecnologico(CNPq) Coordenacao de Aperfeicoamento de Pessoal deNıvel Superior (CAPES) and Fundacao de Amparo aPesquisa do Estado do Rio de Janeiro (FAPERJ) for financialsupport and scholarships to AEP VPO and NTM

References

[1] L Kirkendale G W Saunders and P Winberg ldquoA molecularsurvey of ulva (chlorophyta) in temperate australia reveals

enhanced levels of cosmopolitanismrdquo Journal of Phycology vol49 no 1 pp 69ndash81 2013

[2] R S Steneck and M N Dethier ldquoA functional group approachto the structure of algal-dominated communitiesrdquo Oikos vol69 no 3 pp 476ndash498 1994

[3] R L Fletcher ldquoThe occurrence of ldquogreen tidesrdquomdash a reviewrdquo inMarine Benthic Vegetation Recent Changes and The Effects ofEutrophication vol 123 of Ecological Studies pp 7ndash43 SpringerBerlin Heidelberg Berlin Germany 1996

[4] T A Nelson K Haberlin A V Nelson et al ldquoEcologicaland physiological controls of species composition in greenmacroalgal bloomsrdquo Ecology vol 89 no 5 pp 1287ndash1298 2008

[5] M Teichberg S E Fox Y S Olsen et al ldquoEutrophication andmacroalgal blooms in temperate and tropical coastal watersnutrient enrichment experimentswithUlva spprdquoGlobal ChangeBiology vol 16 no 9 pp 2624ndash2637 2010

[6] R Paulert V Talamini J E F CassolatoM E R Duarte M DNoseda and M J Stadnik ldquoEffects of sulfated polysaccharideand alcoholic extracts from green seaweed Ulva fasciata onanthracnose severity and growth of common bean (Phaseolusvulgaris L)rdquo Journal of Plant Diseases and Protection vol 116no 6 pp 263ndash270 2009

[7] M Lenzi G Salvaterra P Gennaro et al ldquoA new approach tomacroalgal bloom control in eutrophic shallow-water coastalareasrdquo Journal of Environmental Management vol 150 pp456endash465e 2015

[8] Z Gao D Xu C Meng et al ldquoThe green tide-formingmacroalgaUlva linza outcompetes the redmacroalga Gracilarialemaneiformis via allelopathy and fast nutrients uptakerdquoAquatic Ecology vol 48 no 1 pp 53ndash62 2014

[9] H Verbruggen ldquoMorphological complexity plasticity andspecies diagnosability in the application of old species namesin DNA-based taxonomiesrdquo Algae Highlights vol 50 no 1 pp26ndash31 2013

[10] M Hiraoka S Shimada Y Serisawa M Ohno and H EbataldquoTwo different genetic strains of stalked-Ulva (Ulvales Chloro-phyta) grow on intertidal rocky shores in Ebisujima centralJapanrdquo Phycological Research vol 51 no 3 pp 161ndash167 2003

[11] D Butler ldquoAncient algal mixup sortedrdquo Nature 2007[12] L G K Kraft G T Kraft and R F Waller ldquoInvestigations into

southern Australian ulva (ulvophyceae chlorophyta) taxonomyand molecular phylogeny indicate both cosmopolitanism andendemic cryptic speciesrdquo Journal of Phycology vol 46 no 6 pp1257ndash1277 2010

[13] C J OrsquoKelly A Kurihara T C Shipley and A R SherwoodldquoMolecular assessment of ulva spp (ulvophyceae chlorophyta)in the hawaiian islandsrdquo Journal of Phycology vol 46 no 4 pp728ndash735 2010

[14] N Knowlton ldquoSibling species in the seardquo Annual Review ofEcology Evolution and Systematics vol 24 no 1 pp 189ndash2161993

[15] P D N Hebert A Cywinska S L Ball and J R DeWaardldquoBiological identifications throughDNA barcodesrdquo Proceedingsof the Royal Society B Biological Science vol 270 no 1512 pp313ndash321 2003

[16] N Puillandre P Bouchet M C Boisselier-Dubayle et al ldquoNewtaxonomy and old collections Integrating DNA barcoding intothe collection curation processrdquo Molecular Ecology Resourcesvol 12 no 3 pp 396ndash402 2012

[17] G Du F Wu Y Mao S Guo H Xue and G Bi ldquoDNAbarcoding assessment of green macroalgae in coastal zone


around Qingdao Chinardquo Journal of Ocean University of Chinavol 13 no 1 pp 97ndash103 2014

[18] W L Bauerle D J Weston J D Bowden J B Dudley and J EToler ldquoLeaf absorptance of photosynthetically active radiationin relation to chlorophyll meter estimates among woody plantspeciesrdquo Scientia Horticulturae (Amsterdam) vol 101 no 1-2pp 169ndash178 2004

[19] N R Baker ldquoChlorophyll fluorescence a probe of photosynthe-sis in vivordquo Annual Review of Plant Biology vol 59 pp 89ndash1132008

[20] U Schreiber W Bilger and C Neubauer ldquoChlorophyll fluores-cence as a nonintrusive indicator for rapid assessment of in vivophotosynthesisrdquo Ecological Studies vol 100 pp 49ndash70 1994

[21] A Espinosa-Calderon I Torres-Pacheco J A Padilla-Medinaet al ldquoDescription of photosynthesis measurement methodsin green plants involving optical techniques advantages andlimitationsrdquo African Journal of Agricultural Research vol 6 no12 pp 2638ndash2647 2011

[22] T P J Kruger C Ilioaia and R Van Grondelle ldquoFluorescenceintermittency from the main plant light-harvesting complexResolving shifts between intensity levelsrdquoThe Journal of PhysicalChemistry B vol 115 no 18 pp 5071ndash5082 2011

[23] A A Fernandez-Jaramillo C Duarte-Galvan L M Contreras-Medina et al ldquoInstrumentation in developing chlorophyllfluorescence biosensing a reviewrdquo Sensors (Switzerland) vol 12no 9 pp 11853ndash11869 2012

[24] V P Oliveira N T Martins Y Yoneshigue-Valentin and AEnrich-Prast ldquoChlorophyll-a fluorescence in ecology theoret-ical considerations and examples around marine macroalgaerdquoJournal of Environmental Science and Engineering vol A22 pp1165ndash1172 2013

[25] K Maxwell and G N Johnson ldquoChlorophyll fluorescencemdashapractical guiderdquo Journal of Experimental Botany vol 51 no 345pp 659ndash668 2000

[26] F Mendez F Tala R Rautenberger et al ldquoMorphologicaland physiological differences between two morphotypes ofdurvillaea antarctica (phaeophyceae) from the sub-antarcticecoregion of magallanes chilerdquo Journal of Applied Phycologyvol 29 no 5 pp 2557ndash2565 2017

[27] W R TaylorMarine algae of the eastern tropical and subtropicalcoasts of the Americas The University of Michigan PressMichigan Mich USA 1960

[28] A B Joly ldquoFloramarinha do litoral norte do estado de sao pauloe regioes circunvizinhasrdquo in Boletim da Faculdade de FilosofiaCiencias e Letras da Universidade de Sao Paulo vol 249 pp 5ndash393 Departamento de Botanica da USP Universidade de SaoPaulo 1965

[29] Y Yoneshigue Taxonomie et ecologie des algues marines dans laregion de cabo frio (rio de janeiro bresil)[PhD thesis] UniversitedrsquoAix-Marseille II Marseille France 1985

[30] Y Yoneshigue-Valentin and J L Valentin ldquoMacroalgae of thecabo frio upwelling region brazil ordination of communitiesrdquoin Coastal plant communities of Latin America U Seeliger EdAcademic Press San Diego Calif USA 1992

[31] H Von StoschWirkungen von Jod und Arsenit auf Meeresalgenin Kultur vol 4 of Proc Inter Seaw Symp 1963

[32] J L Valentin D L Andre and S A Jacob ldquoHydrobiology inthe cabo frio (Brazil) upwelling two-dimensional structure andvariability during a wind cyclerdquo Continental Shelf Research vol7 no 1 pp 77ndash88 1987

[33] B Kjerfve C H A Ribeiro G T M Dias A M Filippoand V D S Quaresma ldquoOceanographic characteristics of animpacted coastal bay baıa de guanabara rio de janeiro BrazilrdquoContinental Shelf Research vol 17 no 13 pp 1609ndash1643 1997

[34] B Genty J M Briantais and N R Baker ldquoThe relationshipbetween the quantumyield of photosynthetic electron transportand quenching of chlorophyll fluorescencerdquo Biochimica etBiophysica Acta (BBA) - General Subjects vol 990 pp 87ndash921989

[35] J C Kromkamp and R M Forster ldquoThe use of variablefluorescence measurements in aquatic ecosystems differencesbetween multiple and single turnover measuring protocols andsuggested terminologyrdquo European Journal of Phycology vol 38no 2 pp 103ndash112 2003

[36] W Han X W Xu L Li J Q Lei and S Y Li ldquoChlorophyll afluorescence responses of Haloxylon ammodendron seedlingssubjected to progressive saline stress in the Tarim deserthighway ecological shelterbeltrdquo Photosynthetica vol 48 no 4pp 635ndash640 2010

[37] M D Guiry and G M Guiry Algae Base Galway IrelandWorldwide electronic publication National University of Ire-land 2018 httpwwwalgaebaseorg

[38] I R Davison ldquoEnvironmental effects on algal photosynthesistemperaturerdquo Journal of Phycology vol 27 no 1 pp 2ndash8 1991

[39] Y Wang Y Wang L Zhu B Zhou X Tang and B NeilanldquoComparative studies on the ecophysiological differences of twogreen tide macroalgae under controlled laboratory conditionsrdquoPLoS ONE vol 7 no 8 Article ID e38245 2012

[40] J A Raven and R J Geider ldquoTemperature and algal growthrdquoNew Phytologist vol 110 no 4 pp 441ndash461 1988

[41] L C Rai and J P Gaur Algal Adaptation to EnvironmentalStresses Physiological Biochemical and Molecular MechanismsSpringer Berlin Heidelberg Berlin Germany 2001

[42] M A De Guimaraes and R Coutinho ldquoSpatial and temporalvariation of benthic marine algae at the cabo frio upwellingregion rio de janeiro Brazilrdquo Aquatic Botany vol 52 no 4 pp283ndash299 1996

[43] I R Davison and G A Pearson ldquoStress tolerance in intertidalseaweedsrdquo Journal of Phycology vol 32 no 2 pp 197ndash211 1996

[44] J L Valentin ldquoAnalyse des parametres hydrobiologiques dansla remontee de cabo frio (Bresil)rdquoMarine Biology vol 82 no 3pp 259ndash276 1984

[45] G C Pereira R Coutinho and N F F Ebecken ldquoData miningfor environmental analysis and diagnostic a case study ofupwelling ecosystem of arraial do cabordquo Brazilian Journal ofOceanography vol 56 no 1 pp 1ndash12 2008

[46] P G Falkowski and J A Raven Aquatic Photosynthesis Prince-ton University Press New Jersey NJ USA 2007

[47] Y B Ho ldquoUlva lactuca (Chlorophyta Ulvales) in Hong Kongintertidal waters ndash its nitrogen and phosphorus contents andits use as a bioindicator of eutrophicationrdquo in Asian MarineBiology vol 4 pp 97ndash102 1987

[48] I DIez A Secilla A Santolaria and J M Gorostiaga ldquoPhyto-benthic intertidal community structure along an environmentalpollution gradientrdquoMarine Pollution Bulletin vol 38 no 6 pp463ndash472 1999

[49] B Worm and H K Lotze ldquoEffects of eutrophication grazingand algal blooms on rocky shoresrdquo Limnology and Oceanogra-phy vol 51 no 1 pp 569ndash579 2006

[50] S Shimada M Hiraoka S Nabata M Iima and M MasudaldquoMolecular phylogenetic analyses of the Japanese Ulva and


Enteromorpha (Ulvales Ulvophyceae) with special reference tothe free-floating Ulvardquo Phycological Research vol 51 no 2 pp99ndash108 2003

[51] MCormaci G Furnari andGAlongi ldquoMarine benthic flora oftheMediterranean sea chlorophytardquoBollettino delle sedute dellaAccademia Gioenia di Scienze Naturali in Catania vol 377 pp11ndash436 2014

[52] K Kostamo J BlomsterH Korpelainen et al ldquoNewmicrosatel-lite markers for ulva intestinalis (chlorophyta) and the transfer-ability ofmarkers across species of ulvaceaerdquoPhycologia vol 47no 6 pp 580ndash587 2008

[53] C J Loughnane L M McIvor F Rindi D B Stengel andM D Guiry ldquoMorphology rbcL phylogeny and distributionof distromatic Ulva (Ulvophyceae Chlorophyta) in Ireland andsouthern Britainrdquo Phycologia vol 47 no 4 pp 416ndash429 2008

[54] L C Hofmann J C Nettleton C D Neefus and A CMathieson ldquoCryptic diversity of Ulva (Ulvales Chlorophyta) inthe great bay Estuarine system (Atlantic USA) introduced andindigenous distromatic speciesrdquo European Journal of Phycologyvol 45 no 3 pp 230ndash239 2010

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Submit your manuscripts atwwwhindawicom


morphological characterization of most studies which hasled to uncertainty and sometimes to misinterpretation (see[1 11])

In the latter context physiological markers that areassociated with the photosynthesis process may be espe-cially helpful Photosynthetic organisms adjust the opera-tion of their photosynthetic apparatus to optimize or pre-serve metabolism under stress which can be detected bychlorophyll-a fluorescence variation [18 19] In this casethe evaluation of the effective quantum yield of PSII bypulse amplitude modulated (PAM) fluorometers has beenproposed as an attractive measure of the ecophysiologicalcondition [20ndash24] This technique is notable for performinga nonintrusive real-time analysis under both laboratory andfield conditions [25] and has been applied for establishing thedifferences among morphotypes of macroalgae [26]

Under this background the aim of this study was to pro-vide comparative ecophysiological information about threecooccurring species of Ulva in the Southwestern Atlantic(Ulva rigida C Agardh Ulva lactuca Linnaeus and Ulvafasciata Delile) that were morphologically identified Weassessed the effect of temperature and nutrient variation onthe photosynthetic efficiency (by chlorophyll-a fluorescencemeasurements) of the three species The general testedhypothesis was that species would have different responses toboth of the tested factors We subsequently expect to provideuseful information for using PAM fluorometry of Ulva asa reliable tool for ecophysiological assessment and for thetaxonomic debate about Ulva

2 Material and Methods

21 Sampling The three species of Ulva (U fasciata U lac-tuca and U rigida) were collected during summer (January2010) in the intertidal zone at Prainha Beach (22∘ 571015840S 042∘011015840W) Rio de Janeiro Brazil Immediately the epibionts wereremoved and the macroalgae were kept in cool boxes withlocal seawater (sim30 120583M NO

3 3 120583M PO

4 19∘C and 35 PSU)

filtered through 10-120583mmesh filters

22 The Species The morphological taxonomy of the threestudied species (U fasciata U lactuca and U rigida) wasbased on botanical descriptions and inventories that consid-ered in-depth confrontation among the species descriptionsand the Southwestern Atlantic specimens [27ndash30] Individ-uals were prepared as herbarium vouchers and depositedin the herbarium of the Universidade Federal do Rio deJaneiroUFRJ (under the accession numbers RFA 4244142442 42443)

23 Experimental Design All the individuals were main-tained in Erlenmeyer flasks in a proportion of 1 g of freshmass Lminus1 pasteurized oligotrophic ocean water with addedtrace nutrients (lt1 120583M NH

4 lt1 120583M NO

3 lt2 120583M NO

2 lt1

120583MPO4) at 20∘C and 200 120583mol photons mminus2 sminus1 (PAR pho-

tosynthetically active radiation) for 15 days This phase wasestablished to set all the macroalgae to the same nutritionaland physiological state before the manipulative experiment

This PAR was chosen because it is an intermediate value inrelation to the saturation parameter that is found in the P-Icurves (100ndash400 photons mminus2 sminus1)

After the nutrient deprivation period six treatments wereapplied using combinations of two temperatures and threeconcentrations of nutrients with 5 replicates per treatmentThe experiment per se lasted 10 hours The factor ldquoTemper-aturerdquo was represented by 20∘C and 30∘C and the factorldquoNutrientrdquo was represented by different additions of NaNO

3

minus

and Na2HPO412H2O of Von Stoschrsquos enriched seawater

(VSE 500 120583MNO3

minus and 30 120583MPO4

minus) [31]The experimentalsetup values of temperatures are related to the temperatureat the upper intertidal fringe at the sampling site whichranges between 18∘C (high tide) and 32∘C (low tide) [29]and the nutrient concentrations were set as 01 VSE 02 VSEand 05 VSE These concentrations refer to a gradient ofnutrient concentrations that were recorded in SouthwesternAtlantic [32 33] The treatment set as 01 VSE refers tonatural eutrophicated waters (upwelling events at collectingsite Arraial do Cabo Rio de Janeiro) [32] whereas 02 VSEand 05 VSE refer to the range of nutrient concentrations thatwere recorded in the human-disturbed Guanabara Bay Riode Janeiro [33]

24 Fluorescence Measurements The photosynthetic effi-ciency of the three species of Ulva in every combinationof temperature and nutrient concentration was charac-terized by chlorophyll-a fluorescence measurements usinga submersible diving-PAM fluorometer (Walz EffeltrichGermany) The measurements consisted of recording theeffective quantum yield of PSII which was calculated as Y= (F1015840m minus Ft)F

1015840

m where F1015840m is the maximum fluorescencein the light (obtained by a saturating actinic light pulse ndash8900 120583mol photons mminus2 sminus1 08 s) and Ft is the steady-state of fluorescence in the light [34 35] Considering thepossibility of physiological differences along the thallus [36]the fluorescence was measured in a similar region of thethallus in each individual

25 Data Analyses To determine the effects if any of tem-perature and nutrients on these three species of Ulva basedon the Y measurements a four-way analysis of variance wasperformedThe analysis included the (i) species (orthogonalfixed three levels) (ii) nutrients (orthogonal fixed three lev-els) (iii) temperature (orthogonal fixed two levels) and (iv)plot Erlenmeyer flasks (random and nested in the interactionof factors 1 and 2 two levels) The SNK post hoc test was usedto examine the nature of the differences that were detectedwith ANOVA (005 significance level) Statistical analyseswere performed using GMAV5 for Windows

3 Results

Differences in the Y were found for the interaction ofspecies and temperature (Table 1) For both U lactuca and Urigida the individuals that were incubated at 30∘C showed ahigher photosynthetic efficiency than did those incubated at20∘C whereas no difference in photosynthetic efficiency was






080

079

078

077

076

075

Y

20∘C 30

∘C 20∘C 30

∘C 20∘C 30




4 Discussion











Data Availability




Acknowledgments


References






























































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018







080

079

078

077

076

075

Y

20∘C 30

∘C 20∘C 30

∘C 20∘C 30




4 Discussion











Data Availability




Acknowledgments


References






























































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018






Data Availability




Acknowledgments


References






























































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018













































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018


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