The Gracilariaceae Germplasm Bank of the University of So Paulo, Brazil-a DNA barcoding approach

The University of So Paulo Gracilariaceae Germplasm Bank has 50 strains collected mostly in Brazil, but also elsewhere in the world. This bank has been used as a source of material for research developed locally and abroad. With over 200 species, some of which have high economic value, the family Gracilariaceae has been extensively studied. Nonetheless, taxonomic problems still persist by the existence of cryptic species, phenotypic plasticity, and broad geographic distribution. In the case of algae kept in culture for long periods of time, the identification is even more problematic as a consequence of considerable morphological modification. Thus, the use of molecular markers has been shown to be an efficient tool to elucidate taxonomic issues in the group. In this work, we sequenced the 5'-end of the cox1 gene for 41 strains and the universal plastid amplicon (UPA) plastid region for 45 strains, covering all 50 strains in the bank. In addition, the rbcL for representatives of the cox1/UPA clusters was sequenced for 14 strains. The original species identification based on morphology was compared with the molecular data obtained in this work, resulting in the identification of 13 different species. Our analyses indicate that cox1 and UPA are suitable markers for the delineation of species of Gracilariales in the germplasm bank. The addition of DNA barcode tags to the samples in the Gracilariaceae germplasm bank and the molecular identification of the species will make this bank even more useful for future research as the species can be easily traced and confirmed.

Molecular support for the establishment of the new genus Laurenciella within the Laurencia complex (Ceramiales, Rhodophyta)

Currently, five genera are assigned to red seaweeds of the Laurencia complex worldwide: Chondrophycus, Laurencia s.s., Osmundea, Palisada and Yuzurua. The genera are segregated on the basis of morphological characters, especially the reproductive traits, and molecular sequences of the plastid-encoded gene rbcL. Four of the genera have been resolved as monophyletic, but not Laurencia s.s. In this study based on an rbcL gene phylogeny we show the presence of a sixth lineage within the Laurencia complex, viz., Laurencia marilzae plus two unidentified species of Laurencia from Brazil. The phylogenetic position of this group, combined with the high genetic divergence from Laurencia s.s. (8.2-11%), strongly support the establishment of a sixth genus for the complex, proposed here as Laurenciella gen. nov. This new taxon differs from Laurencia s.s. and from the other genera of the complex by molecular sequence data, but is indistinguishable from Laurencia s.s. by the usual morphological features.

Redefining the taxonomic status of Laurencia dendroidea (Ceramiales, Rhodophyta) from Brazil and the Canary Islands

Morphological and molecular studies have been performed on Laurencia dendroidea derived from Brazil and the Canary Islands. This species possesses all of the characters that are typical of the genus Laurencia, including the production of the first pericentral cell underneath the basal cell of the trichoblast; the production of tetrasporangia from particular pericentral cells without the formation of additional fertile pericentral cells; spermatangial branches that are produced from one of two laterals on the suprabasal cell of the trichoblasts; and a procarp-bearing segment that possesses five pericentral cells. The phylogenetic position of L. dendroidea was inferred by analysing the chloroplast-encoded rbcL gene sequences of 51 taxa. Phylogenetic analyses revealed that the taxa previously identified and cited in Brazil as Laurencia filiformis, L. majuscula and L. obtusa and in the Canary Islands as L. majuscula all represent the same taxonomic entity and examination of type material allowed us to identify this entity as L. dendroidea, whose type locality is in Brazil. Laurencia obtusa from the Northern Atlantic is confirmed to represent a distinct species, which displays high genetic divergence with respect to western and eastern Atlantic samples. The phylogenetic analyses also supported the nomenclatural transfer of Chondrophycus furcatus (Cordeiro-Marino & M. T. Fujii) M. T. Fujii & Senties to Palisada furcata (Cordeiro-Marino & M. T. Fujii) Cassano & M. T. Fujii comb. nov.

Morphological and molecular evidence demonstrates the amphi-Atlantic distribution of Laurencia catarinensis (Ceramiales, Rhodophyta)

We performed morphological and molecular studies of Laurencia catarinensis from the Canary Islands. This species has an entangled habit, cushion-like tuft formation, cortical cell walls, slightly to markedly projecting near the apex, and lacking lenticular thickenings in medullary cells. We inferred its phylogenetic position by analyzing the chloroplast-encoded rbcL gene sequences from 41 samples. The results demonstrate that specimens of L. catarinensis from the Canary Islands, where it is referred to as L. intricata, and those from Brazil (including specimens from the type locality in Santa Catarina) form a monophyletic clade with low genetic divergence (0-0.9%). In contrast, specimens of L. intricata from the type locality in Cuba, Mexico, and the USA were clearly distinct from L. catarinensis collected in Brazil and the Canary Islands, as shown by high genetic divergence values (4.9-5.7%). The type material of L. catarinensis from Brazil allowed us to identify all samples from the Canarian Archipelago as L. catarinensis. These findings expand the known geographical distribution of L. catarinensis to the eastern Atlantic Ocean and demonstrate an amphi-Atlantic distribution of the species.

Morphological and molecular evidences within Osmundea (Ceramiales, Rhodophyta) from the Canary Islands, eastern Atlantic Ocean

The genus Osmundea is a strongly supported monophyletic group within the Laurencia complex and shows a disjunct distribution occurring in the North-East and South-West Pacific, the Indian and Atlantic oceans and the Mediterranean Sea. Its phenotypic plasticity on the Canary Islands may be the result of the high ecological variability partially due to the particular oceanographic characteristics in this region. The combination of molecular analyses based on the comparison of the chloroplast-encoded rbcL sequences and morphological data allowed us to delimit three distinct taxa from the coasts of the Canarian Archipelago: Osmundea pinnatifida, Osmundea truncata and an unidentified species, Osmundea sp. Moreover, the high value of genetic divergence between Osmundea sp. and the rest of the Osmundea species suggests that this taxon should be assigned to a new species within the Osmundea genus. Occurrence of O. hybrida and O. oederi (synonym: O. ramosissima) has not been confirmed. Our results also suggest a possibly questionable record of the taxa O. hybrida and O. oederi on the Canary Islands.

Is Drosera meristocaulis a pygmy sundew? Evidence of a long-distance dispersal between Western Australia and northern South America

South America and Oceania possess numerous floristic similarities, often confirmed by morphological and molecular data. The carnivorous Drosera meristocaulis (Droseraceae), endemic to the Neblina highlands of northern South America, was known to share morphological characters with the pygmy sundews of Drosera sect. Bryastrum, which are endemic to Australia and New Zealand. The inclusion of D. meristocaulis in a molecular phylogenetic analysis may clarify its systematic position and offer an opportunity to investigate character evolution in Droseraceae and phylogeographic patterns between South America and Oceania. was included in a molecular phylogenetic analysis of Droseraceae, using nuclear internal transcribed spacer (ITS) and plastid rbcL and rps16 sequence data. Pollen of D. meristocaulis was studied using light microscopy and scanning electron microscopy techniques, and the karyotype was inferred from root tip meristem. The phylogenetic inferences (maximum parsimony, maximum likelihood and Bayesian approaches) substantiate with high statistical support the inclusion of sect. Meristocaulis and its single species, D. meristocaulis, within the Australian Drosera clade, sister to a group comprising species of sect. Bryastrum. A chromosome number of 2n approx. 3236 supports the phylogenetic position within the Australian clade. The undivided styles, conspicuous large setuous stipules, a cryptocotylar (hypogaeous) germination pattern and pollen tetrads with aperture of intermediate type 78 are key morphological traits shared between D. meristocaulis and pygmy sundews of sect. Bryastrum from Australia and New Zealand. The multidisciplinary approach adopted in this study (using morphological, palynological, cytotaxonomic and molecular phylogenetic data) enabled us to elucidate the relationships of the thus far unplaced taxon D. meristocaulis. Long-distance dispersal between southwestern Oceania and northern South America is the most likely scenario to explain the phylogeographic pattern revealed.

Phylogenetic analysis of Codium species from Brazil, with the description of the new species C. pernambucensis (Bryopsidales, Chlorophyta)

The genus Codium comprises c. 125 species widely distributed in marine coastal environments throughout the world. Due to morphological plasticity, the taxonomic delimitation of Codium species can be difficult. Sequences of the first exon of the large subunit of RUBISCO (rbcL) have been used in the molecular delimitation of species and for phylogenetic purposes. In the present study, we complement previous morphological work on Brazilian Codium species with molecular systematics. Based on the partial rbcL sequences, seven species are recognized along the Brazilian coast: C. decorticatum, C. intertextum, C. isthmocladum, C. profundum, C. spongiosum, C. taylorii and the new species Codium pernambucensis. Ten unique sequences were obtained among the samples examined, which we used in combination with previously published sequences to infer molecular phylogenies using various methods. The resulting trees showed three principal monophyletic groupings: Clade A with species having a prostrate habit, not branched, and mostly with small, grouped utricles; Clade B primarily consisting of upright species with cylindrical branches and large individual utricles; and Clade C composed of upright species with cylindrical branches that are slightly flattened, and have intermediate-sized individual utricles. The Brazilian species grouped with morphologically similar taxa from other geographic localities, and are present in all three main clades. A new sprawling species, Codium pernambucensis is described based on morphology and molecular analyses.

Morphological and molecular studies on the Brazilian native red seaweed Laurencia oliveirana (Rhodomelaceae, Ceramiales)

Morphological and molecular studies were carried out on Laurencia oliveirana from the type locality (Arraial do Cabo, Rio de Janeiro, Brazil). This species is easily recognized by its small size, sub-erect habit forming intricate cushion-like tufts and unilateral pectinate branching. The species displays all the typical characters of the genus Laurencia, such as the production of the first pericentral cell underneath the basal cell of the trichoblast, tetrasporangia produced from particular pericentral cells, with the third and fourth pericentral cells becoming fertile, without production of additional pericentral cells, spermatangial branches produced from one of two laterals on the suprabasal cell of trichoblasts, and procarp-bearing segment with five pericentral cells. Details of tetrasporangial plants and development of procarp and male plants are described for the first time for the species. The phylogenetic position of L. oliveirana was inferred by analysis of the chloroplast-encoded rbcL gene sequences from 57 taxa. In all phylogenetic analyses, L. oliveirana grouped with L. caraibica, L. caduciramulosa, L. venusta and L. natalensis, forming a monophyletic clade within the Laurencia sensu stricto. The genetic divergence between L. oliveirana and the molecularly closest species, L. caraiba collected in Brazil, was 2.3%.