a dna barcode approach of the laurencia complex ......support, represented by the taxa: l. marilzae,...
TRANSCRIPT
RESULTS
DISCUSSION
1Dpto. Botânica, Universidade de São Paulo, São Paulo, Brazil, [email protected], 2Dpto. Biología Vegetal (Botánica). Univ. de La Laguna, Canary Islands, Spain, 3CIIMAR, CIRN & Departamento de Biologia, Universidade dos Azores, Ponta Delgada, Azores, Portugal, 4Dpto. Hidrobiología, Univ. Autónoma Metropolitana,
Iztapalapa, México, D.F., 5Departament of Biological Sciences, Florida International University, Miami, Florida, USA, 6Seção de Ficologia, Instituto de Botânica, São Paulo, Brazil.
The diversity of the Laurencia complex is being assessed in tropical and
subtropical Atlantic by an international cooperation project involving
Brazil, Mexico, Spain (Canary Islands), Portugal (Azores and Madeira)
and USA (Florida) on the base of molecular data allied to a detailed
morphological study of species. The diversity of the complex was
analyzed for the first time for the Atlantic Ocean, including specimens
from all five localities, using the plastid 23S rRNA gene (UPA), which has
been investigated as potential DNA Barcode marker for photosynthetic
eukaryotes. The COI-5P gene was also used as DNA barcode, and the
rbcL gene was used for phylogenetic inferences.
MATERIAL AND METHODSINTRODUCTION
A DNA BARCODE APPROACH OF THE LAURENCIA COMPLEX (CERAMIALES, RHODOPHYTA) IN THE TROPICAL AND SUBTROPICAL ATLANTIC OCEAN
Valéria Cassano1, Maria Machín-Sánchez2, Ana I. Neto3, Mariana C. Oliveira1, Abel Sentíes4, Jhoana Díaz-Larrea4, Mª Candelaria Gil-Rodríguez2, Ligia Collado-Vides5,
Amanda Medeiros1, Alain Duran5 & Mutue T. Fujii6
Chondria dangeardii Hawaii (HQ421425)C. arcuata Hawaii (HQ421424)
L. majuscula Hawaii (HQ421434)L. dendroidea Rio de Janeiro, Brazil L. dendroidea Rio de Janeiro, Brazil
L. dendroidea Bahia, Brazil L. dendroidea Bahia, BrazilL. dendroidea Bahia, Brazil
L. dendroidea Espírito Santo, BrazilL. dendroidea Rio de Janeiro, BrazilL. dendroidea Rio de Janeiro, Brazil
I L. dendroidea São Paulo, BrazilL. nidifica Hawaii (HQ421001)
L. viridis Canary Islands, Spain (MMS0094)L. cf. pyramidalis Azores, Portugal (PIX-11-26)L. cf. pyramidalis Azores, Portugal (SMG-11-72)L. pyramidalis Canary Islands, Spain (MMS0096)L. pyramidalis Azores, Portugal (PIX-11-11)L. mcdermidiae Hawaii (HQ4214744)L. brachyclados Hawaii (HQ421426)L. galtsoffii Hawaii (HQ421136)
L. decumbens Hawaii (HQ420991)L. oliveirana Rio de Janeiro, Brazil
L. caraibica Rio Grande do Norte, BrazilL. catarinensis Santa Catarina, Brazil L. catarinensis Rio de Janeiro, Brazil L. catarinensis Espírito Santo, Brazil
L. crustiformans Hawaii (HQ421480)P. parvipapillata Hawaii (HQ421473)
P. yamadana Havaí (HQ421478)P. perforata São Paulo, Brazil (IBC0019)
P. perforata Bahia, BrazilP. perforata Canary Islands, Spain (MMS0093)P. perforata São Paulo, Brazil (IBC00165)P. perforata São Paulo, Brazil (IBC00103)P. perforata São Paulo, Brazil (IBC0034)P. perorata Espírito Santo, Brazil
P. flagellifera Canary Islands, Spain (MMS0095)P. flagellifera Espírito Santo, Brazil
P. flagellifera Rio de Janeiro,BrazilP. flagellifera Bahia, Brazil
Chondrophycus cartilagineus Hawaii (HQ421430)P. furcata Bahia, Brazil
L. cervicornis Florida, USAO. sp. 3 Azores, Portugal (PIX-11-12)
O. truncata Canary Islands, SpainO. truncata Canary Islands, Spain (MMS0098)
O. sp. 7 Azores, Portugal (PIX-11-44)O. sp. 7 Azores, portugal (PIX-11-13)
O. sp. 4 Azores, Portugal (PIX-11-133)O.cf. pinnatifida Azores, Portugal (PIX-11-31)O. cf. pinnatifida Azores, Portugal (PIX-11-82)
O. sp. 5 Azores, Portugal (SMG-11-78)C. succisus Hawaii (HQ421477)
C. dotyi Hawaii (HQ421432)
C. undulatus Hawaii (HQ421208)
L. translucida Bahia, BrazilL. translucida Bahia, Brazil
L. sp.1 São Paulo, Brazil (IBC003)L. sp. 1 São Paulo, Brazil (IBC0014)L. sp. 1 Espírito Santo, Brazil
L. sp. 1 Espírito Santo, BrazilL. cf. marilzae Azores, Portugal (PIX-11-55)L. marilzae Canary Islands, Spain (MMS0097)L. marilzae São Paulo, Brazil
0.005 substitutions/site
100
99
86
100
96
86
9197
73
73
80
90
83
78
76
71
Laurencia s.s.
Palisada
Osmundea
Chondrophycus
Laurenciella
Fig. 1. Neighbor-Joining analysis for UPA sequences for the Laurenciacomplex. The bootstrap values for 2000 replicates are shown on thebranches (only values above 70 were considered).
Markers/ divergences (%) rbcL COI-5P UPA
Intergeneric 5.6-11.9 6.9-13.1 3.1-7.8
Interspecific 1.9-6.2 4.0-9.3 0.8-2.9
Intraspecific 0-1 0-2.5 0-0.5
Tab. 1. Range of genetic divergence among DNA sequences of the Laurenciacomplex for different markers. Problematic taxa were excluded from thecomparison.
The genus Laurenciella established based on rbcL sequences was also confirmed with theuse of two other markers: UPA and COI-5P, forming independent clades with highsupport, represented by the taxa: L. marilzae, L. sp. 1 and L. sp. 2. The intergenericdivergence between Laurencia and Laurenciella for UPA and COI-5P was in the range ofvariation obtained from other genera of the complex, 4.9-5.8% and 10.1-13.1%,respectively.
Laurencia cervicornis from Florida joined with Palisada by the UPA and rbcL markers andits taxonomic position will also have to be better investigated.
The ‘problematic’ Laurencia translucida, an endemic species from Brazil, remains anenigmatic species. Its taxonomic position has always been controversial since it shares acombination of morphological characters to the genera Chondrophycus and Laurencia.Unlike the results with the rbcL gene, L. translucida, was positioned within theChondrophycus clade by UPA and COI-5P markers. The rbcL sequence available of L.translucida seems to be chimeric. New rbcL sequences are necessary to confirm itstaxonomic position.
In the analyses with the UPA and COI-5P, C. cartilagineus, type species the genusChondrophycus, joined with Palisada, which indicates that these two genera can becongeneric. Further analyses are necessary to clarify the position of these taxa.
Financial support
v
Chondria dangeardii USA (GU223879)Palisada sp. (as P. papillosa) MexicoP. perforata Spain P. perforata (as P. papillosa) SpainP. perforata Canary Islands, Spain (MMS0093)
P. perforata Parati, Rio de Janeiro, BrazilP. perforata (as P. papillosa) Rio de Janeiro, BrazilP. perforata (as P. papillosa) Angra dos Reis, BrazilP. perforata Parati, Rio de Janeiro, BrazilP. perforata (as P. papillosa) Rio de Janeiro, BrazilP. perforata São Paulo, Brazil (IBC00103)P. perforata São Paulo, Brazil (IBC0034)P. perforata São Paulo, Brazil (IBC0019)
P. flagelllfera Rio de Janeiro, Brazil P. flagelllfera Rio de Janeiro, Brazil
P. furcata Paraíba, BrazilP. parvipapillata USA (GU223895)
C.cartilagineus USA (GU223896)Y. poiteaui var. gemmifera Mexico
Y. poiteaui var. gemmifera MexicoL. sp. 1 São Paulo, Brazil (IBC0014)L. sp. 1 São Paulo, Brazil (IBC0003)L. sp. 1 São Paulo, Brazil L. sp. 1 São Paulo, Brazil (IBC0004)
L. sp. 2 Espírito Santo, Brazil L. marilzae São Paulo, Brazil
L. cf. marilzae Azores, Portugal (PIX-11-55L. translucida Espírito Santo BrazilC. cf. undulatus USA (GU223886)
C. succisus USA (GU223884)Chondrophycus sp. USA (GU223885)
O.cf. pinnatifida Azores, Portugal (PIX-11-31)L. dendroidea Bahia, Brazil
L. dendroidea Rio de Janeiro, BrazilL. dendroidea Rio de Janeiro, BrazilLL. dendroidea Rio de Janeiro, BrazilL. dendroidea Rio de Janeiro, BrazilL. dendroidea Rio de Janeiro, BrazilL. dendroidea São Paulo, Brazil (IBC0113)L. dendroidea Rio de Janeiro, BrazilL. dendroidea Rio de Janeiro, BrazilL. dendroidea São Paulo, Brazil (IBC0013)L. dendroidea Rio de Janeiro, BrazilL. dendroidea Rio de Janeiro, BrazilL. dendroidea São Paulo, Brazil (IBC0092)
L. dendroidea Rio de Janeiro, BrazilL. dendroidea Rio de Janeiro, BrazilL. dendroidea Rio de Janeiro, BrazilL. dendroidea Rio de Janeiro, Brazil
Laurencia majuscula USA (GU223887) L. aldingensis Espírito Santo, Brazil L. aldingensis Rio de Janeiro, Brazil
L. catarinensis Espírito Santo, BrazilL. catarinensis) Rio de Janeiro, BrazilL. catarinensis Rio de Janeiro, BrazilL. catarinensis (as L. intricata) Rio de Janeiro, BrazilL. catarinensis Rio de Janeiro, BrazilL. catarinensis São Paulo, Brazil (IBC0005)L. catarinensis Santa Catarina, Brazil
L. catarinensis Rio Grande do Norte, BrazilL. catarinensis Canary Islands, Spain
L. catarinensis Canary Islands, SpainL. mcdermidiae USA (GU223877)
L. oliveirana Rio de Janeiro, BrazilL. nipponica Japan (GU223875)L. nipponica Russia (GU223874)
L. caduciramulosa Rio de Janeiro, BrazilL. caduciramulosa Rio de Janeiro, Brazil
L. caduciramulosa Canary Islands,SpainL. sp. Hawaii GU223894
L. nidifica USA (GU223888)Laurencia sp.1 Espírito Santo, BrazilLaurencia sp.1 Espírito Santo, Brazil
L. intricata CubaL. viridis Canary Islands, Spain (MMS0094)
L. pyramidalis Canary Islands, Spain (MMS0096)L. cf. pyramidalis Azores, Portugal (PIX-11-11)
0.005 substitutions/site
72
92
100
100
100
10072
100
100
99
98
100
100
100
80
8872
87
100
100
100
10078
100
10083
100
75
Laurencia s.s.
Chondrophycus
Laurenciella
Palisada
Yuzurua
Osmundea
Fig. 2. Neighbor-Joining analysis for COI-5P sequences for the Laurenciacomplex. The bootstrap values for 2000 replicates are shown on the branches(only values above 70 were considered).
Laurenciella sp. 1
Brazil
Laurenciella sp. 2
Brazil
Laurencia sp. 1
Brazil
Laurenciella cf. marilzae
Azores
Laurencia cf. pyramidalis
Azores
Laurenciella marilzae
Canary Inlands
“Laurencia cervicornis”
Florida
“Laurencia translucida”
Brazil
The UPA gene showed to be more conserved, however, the same genetic groups wereresolved with each of the three markers.
Fig. 3. Consensus tree derived from Bayesian analyses of rbcL sequences. The posterior probabilities are shown as thicker branches. Bootstrap supports for MP/NJ (2000 replicates) are shown at the nodes (only values above 70 were considered); *indicates bootstrap support =100%.
Samples of the Laurencia complex, collected in Brazil, Portugal (Azores and
Madeira), Spain (Canary Islands), Mexico (Caribbean Sea) and Florida
(USA), were sequenced using the markers Universal Plastid Amplicon (UPA),
COI-5P and rbcL. For COI-5P and UPA were performed neighbor-joining (NJ)
analyses using PAUP 4.0b10. For rbcL, the phylogenetic relationships were
inferred with PAUP 4.0b10 and MrBayes v.3.0 beta 4. The range of genetic
divergence for the markers used was calculated using "uncorrected 'p'
distance with PAUP.
v
Chondria collinsiana Brazil (GU330225)C. dasyphylla USA (U04021)
L. sp. 3 São Paulo, Brazil (IBC003)L. sp. 3 São Paulo, Brazil (IBC014)L. sp. 3 São Paulo, Brazil L. sp. 3 São Paulo, Brazil (IBC004)L. sp. 2 Espírito Santo, BrazilL. marilzae Brazil (GU938189)
L. cf. marilzae Azores, Portugal (PIX-11-55)Y. poiteuai var. gemmifera Cuba (EF061650)Y. poiteaui var. poiteaui USA (EF061652)Y. poiteaui var. poiteaui Mexico (EF061653)Yuzurua poiteuai var. gemmifera Mexico (EF061648)
“Laurencia” sp. 2 Canary Islands, Spain (MMS0088)L. dendroidea Bahia ,Brazil (IBC0092)L. dendroidea Rio de Janeiro, Brazil (GU330233) L. dendroidea Bahia ,Brazil (GU330228)
L. dendroidea Canary Islands, Spain (MMS0082)L. sp.1 Espírito Santo, BrazilL. sp.1 Rio de Janeiro, Brazil
L. intricata Cuba (GU330238)L. sp. 5 Canary Islands, Spain (MMS0069)L. sp. 4 Canary Islands, Spain (MMS0085)
L. sp. 3 Canary Islands, Spain (MMS0090)L. obtusa Ireland (AF281881)
L. viridis Azores, Portugal (SMA-11-06)L. viridis Canary Islands, Spain (EF685999)L. cf. pyramidalis Azores, Portugal (SMG-11-72)L. cf. pyramidalis Azores, Portugal (PIX-11-11)L. pyramidalis France (FJ785316)L. cf. pyramidalis Azores, Portugal (PIX-11-50)L. cf. pyramidalis Canary Islands, Spain (MMS0080)
L. caduciramulosa Rio de Janeiro, BrazilL.caduciramulosa Canary Islands , Spain (JF 781525)L. venusta Mexico (EF 061655)
L. oliveirana Rio de Janeiro, Brazil (JF810352)L. natalensis South Africa (AF465816)
L. flexuosa South Africa (AF465815)L. brongniartii Australia (EF061654)
L. aldingensis Espírito, Brazil L. aldingensis Rio de Janeiro, Brazil (JF810351)L. catarinensis Espírtito Santo, BrazilL. catarinensis São Paulo, Brazil (IBC0005)L. catarinensis Santa Catarina, BrazilL. catarinensis Rio de Janeiro, BrazilL
L. catarinensis Rio Grande do Norte, BrazilL. catarinensis Canary Islands, SpainL. catarinensis Canary Islands, Spain
L. cf. nidifica New Caledonia (FJ785315)L. cf. mcdermidiae New Caledonia (FJ785314)
L. cf. mariannensis New Caledonia (FJ785313)L. cf. kuetzingii New Caledonia (FJ785322)
L. translucida Espírito Santo, Brazil (AY588408)O. cf. truncata Azores, Portugal (SMG-11-59)O. cf. pinnatifida Madeira, Portugal (MAD-11-28)
O. cf. pinnatifida Azores, Portugal (PIX-11-82)O. pinnatifica France (AF259495)O. cf. pinnatifida Azores, Portugal (PIX-11-31)O. pinnatifida Canary Islands, Spain
O. osmunda Ireland (AF281877)O. sp. 1 Madeira, Portugal (MAD-11-126)
O. sp. 7 Azores, Portugal (SMG-11-77)O. sp. 7 Azores, Portugal (SMG-11-63)O. sp. 7 Azores, Portugal (SMG-11-70)
O. oederi Ireland (AF281880)O. sp. 5 Azores, Portugal (PIX-11-72)O. sp. 5 Azores, Portugal (SMG-11-74)
O. sp. 6 Madeira, Portugal (MAD-11-29)O. sp. 2 Madeira, Portugal (MAD-11-90)
O. truncata Ireland (AF281879)O. truncata Canary Islands, Spain
O. sinicola USA (AY588407)O. blinksii USA (AY172575)
O. splendens Mexico (AY172576 ) O. spectabilis var. spectabilis Mexico (AY172574)
O. sanctarum São Paulo, BrazilLaurencia coelenterata Cuba
“P. papillosa” Mexico (AY588409)P. perforata Canary Islands, Spain (EU256329)P. perforata Rio de Janeiro, Brazil (EU256330)P. perforata São Paulo, Brazil (IBC0019)
P. flagellifera Rio de Janeiro, Brazil (GU330227)P. furcata Paraíba, Brazil (GU330226 )
Laurencia cervicornis Florida, USAC. cf. undulatus New Caledonia (FJ785307)
C. sp. 1 New caledonia (FJ785309)C. sp. 2 New caledonia (FJ785310 )
C. sp. 3 (FJ785311 )
0.005 substitutions/site
Laurenciella
Yuzurua
Laurencia s.s.
Osmundea
Chondrophycus
Palisada
94/90
*
*
100/99
100/99
97/97
100/96
*
**
*
100/99*
99/90
91/9286/82
100/92*
*
**
*
83/73
*
80/90
*
*
100/96
100/99
*100/98
75/79
*
*
95/88
0.09 – 1.00
40
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P12 – Relative Contribution of Environmental and Spatial Processes in Structuring Stream Macroalgal Communities C. Z. Branco1, P. C. Bispo1, C. K. Peres2, A. F. Tonetto1 and L. H. Branco1 1São Paulo State University - UNESP, Brazil; 2Federal University of Latin American Integration – UNILA, Brazil
P13 – New Insights in the Diversity of the Genus Lobophora (Dictyotales, Phaeophyceae) Based on Molecular and Morphological Evidence O. Camacho1, T. Sauvage1, W. Schmidt1, D. W. Freshwater2 and S. Fredericq1 1University of Louisiana at Lafayette, USA; 2University of North Carolina Wilmington, USA
P14 – A DNA Barcode Approach of the Laurencia Complex (Ceramiales, Rhodophyta) in the Tropical and Subtropical Atlantic Ocean V. Cassano1, M. Machín-Sánchez2, A. I. Neto3, M. C. Oliveira1, A. Sentíes4, J. Díaz-Larrea4, M. Gil-Rodríguez2, L. Collado-Vides5, A. Medeiros1 and M. T. Fujii6 1University of São Paulo, Brazil; 2University of La Laguna, Spain; 3University of the Azores, Portugal; 4Autonomous Metropolitan University, Mexico; 5Florida International University, USA; 6Institute of Botany, Brazil
P15 – Characterization of Batrachospermum gelatinosum (L.) De Candolle and B. arcuatum Kylin (Batrachospermales, Rhodophyta) from the Iberian Peninsula I. S. Chapuis1, M. O. Paiano2, M. Aboal3, P. M. Sánchez Castillo1, O. J. Necchi2, and M. M. Elmosallamy4 1University of Granada, Spain; 2UNESP - Campus de São José do Rio Preto, Brazil; 3University of Murcia, Spain; 4Ain Shams Universit, Egypt
P16 – Present Day Collections of Freshwater Red Algae (Batrachospermales, Rhodophyta) from Historically Important Sites in France W. B. Chiasson, E. D. Salomaki, and M. L. Vis Ohio University, USA
P17 – Morphology, Phylogenetic Relationships and DNA Barcoding of the Bangiales (Rhodophyta) from King George Island, Antarctic and its Adjacent Waters H. G. Choi1, S. M. Kim1, J. H. Kim1 and M. S. Hwang2 1Korea Polar Research Institute, Republic of Korea; 2National Fisheries Research and Development Institute, Republic of Korea P18 – Phycological Educational Endeavors: Assessing Algal Knowledge in Museums, Zoos, Aquariums, and Herbariums J. L. Collier1, R. Fitch2, J. Jorve3, R. Kodner4, J. F. Muhlin5 and K. Schoenrock6 1Stony Brook University, USA; 2Wenatchee Valley College, USA; 3University of British Columbia, Canada; 4Western Washington University, USA; 5Maine Maritime Academy, USA; 6University of Alabama at Birmingham, USA
P19 – Algal Turf Scrubbers: Periphyton Production and Nutrient Recovery on a South Florida Citrus Farm P. E. D'Aiuto1, T. J. Evens2, J.P Albano1 and J.M. Patt1 1United States Department of Agriculture, USA; 2Algal Solutions, LLC, USA
68
western Atlantic and Red Sea specimens indicate newly found diversity representing four distinct species, with one unreported species each for Caribbean Colombia, Caribbean Panama, the NW Gulf of Mexico, and Egypt. These taxa in all likelihood correspond to new species. In addition, we propose range extensions for previously unnamed Lobophora spp. reported in Sun et al. 2012. Three recently collected species from the Red Sea are conspecific with recently characterized taxa from Japan, Palau and Malaysia, and one species from the NW Gulf of Mexico is conspecific with a sample from Curaçao in the Lesser Antilles. The morphological evidence for describing the new species of Lobophora will be discussed in light of the molecular-based results.
43 A DNA Barcode Approach of the Laurencia Complex (Ceramiales, Rhodophyta) in the Tropical and Subtropical Atlantic Ocean V. Cassano1, M. Machín-Sánchez2, A. I. Neto3, M. C. Oliveira1, A. Sentíes4, J. Díaz-Larrea4, M. Gil-Rodríguez2, L. Collado-Vides5, A. Medeiros1 and M. T. Fujii6 ([email protected]) 1University of São Paulo, Brazil; 2University of La Laguna, Spain; 3University of the Azores, Portugal; 4Autonomous Metropolitan University, Mexico; 5Florida International University, USA; 6Institute of Botany, Brazil
The diversity of the Laurencia complex is being assessed in tropical and subtropical Atlantic by an international cooperation project involving Brazil, Mexico, Spain (Canary Islands), Portugal (Azores and Madeira) and USA (Florida) on the base of molecular data allied to a detailed morphological study of species. The diversity of the complex was analyzed for the first time for the Atlantic Ocean, including specimens from all five localities, using the plastid 23S rRNA gene (UPA) which has been investigated as potential DNA Barcode marker for photosynthetic eukaryotes. The mitochondrial cytochrome c oxidase I gene (COI-5P) was also used as DNA barcode for the same set of species, and the rbcL gene was used for phylogenetic inferences. The range of genetic variation was compared for the three markers. The UPA proved to be more conserved; however, the same genetic groups were resolved with each of the three markers confirming the six genera currently established for the complex: Chondrophycus, Laurencia, Laurenciella, Palisada, Osmundea and Yuzurua.
44 Characterization of Batrachospermum gelatinosum (L.) De Candolle and B. arcuatum Kylin (Batrachospermales, Rhodophyta) from the Iberian Peninsula I. S. Chapuis1, M. O. Paiano2, M. Aboal3, P. M. Sánchez Castillo1 and O. J. Necchi2 ([email protected]) 1University of Granada, Spain; 2UNESP - Campus de São José do Rio Preto, Brazil; 3University of Murcia, Spain
Freshwater red algae diversity in the Iberian Peninsula (Spain and Portugal) has been poorly studied. The purpose of this study is to approach the morphological and genetic variation of two most common members of the Batrachospermales in the study area, to better understand their biogeographic and phylogenetic relationships in a more global context. We compared genetically six populations each of B. gelatinosum and B. arcuatum from eight different river basins, using three molecular markers to evaluate genetic diversity: RuBisCo large subunit (rbcL) (fully sequenced at the moment), cytochrome oxidase 2-3 spacer (cox2-3) and the barcode region of cytochrome oxidase I (cox1) (preliminary data available). For the morphological comparison nine additional populations were included in the analysis. A wide morphological variation was observed for most vegetative and reproductive characters. rbcL sequences showed a relatively low genetic divergence: 98.8-100% for B. gelatinosum and 99.9-100% for B. arcuatum. We found no correlation between genetic diversity and morphological variation among the populations of both species. Some taxonomic characters are reevaluated aiming at a more reliable characterization of these species.
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