Estudos sobre a diversidade de macroalgas de manguezais no Brasil tem-se baseado apenas em abordagens morfológicas, nas quais os caracteres empregados são instáveis e pouco informativos para a identificação e delimitação de espécies. Neste contexto, macroalgas de manguezais foram investigadas pela primeira vez no litoral brasileiro usando uma abordagem molecular, tendo como alvo de estudo o manguezal da Ilha Barnabé, Baixada Santista, São Paulo. Foram utilizados marcadores moleculares do tipo Barcode, UPA e COI-5P, além do rbcL, amplamente empregado para inferências filogenéticas, e o SSU rDNA. Além desses, os marcadores ITS e tufA foram empregados exclusivamente para as algas verdes, este último sem sucesso. Quinze espécies foram registradas para a área estudada, sendo dez Rhodophyta e cinco Chlorophyta. Destas, duas não novas ocorrências para o Estado de São Paulo, Caloglossa apomeiotica e Boodleopsis vaucherioidea. Das quatro espécies do gênero Bostrychia identificadas neste estudo: \"Bostrychia calliptera\", B. montagnei, B. moritziana e B. radicans, apenas B. montagnei revelou-se uma espécie molecular e morfologicamente bem definida. As demais formaram complexos de espécies com linhagens moleculares distintas. Para B. radicans e B. moritziana as análises relevaram três e duas linhagens moleculares, respectivamente, das sete identificadas para o complexo B. radicans/B. moritziana na literatura. O táxon identificado como \"B. calliptera\" mostrou alta divergência molecular com sequências de B. calliptera do Brasil, apresentando morfologia \"B. pinnata\", um táxon atualmente reduzido a sinônimo de B. calliptera. Espécies do gênero Caloglossa, excetuando C. ogasawaraensis, são de difícil identificação morfológica devido aos tênues caracteres considerados de valor diagnóstico e a sua considerável plasticidade fenotípica. Caloglossa apomeiotica, C. confusa e C. leprieurii foram identificadas essencialmente com o emprego de marcadores moleculares. Caloglossa apomeiotica pode ser segregada das demais pela presença de biesporângios, o que impossibilita uma identificação morfológica segura quando coletados talos inférteis, enquanto C. confusa possui nós fortemente contritos. Os dados moleculares obtidos para Catenella caespitosa a partir de sequências de SSU sugerem que as citações dessa espécie para o litoral brasileiro podem estar equivocadas já que apresentam alta divergência intraespecífica com C. caespitosa do banco de dados. A falta de sequências da localidade tipo, de sequências com marcadores do tipo Barcode e de uma maior amostragem molecular das espécies de Catenella nos bancos de dados, nos impossibilitaram chegar a um resultado conclusivo. A obtenção de sequências para as algas verdes foi extremamente problemática, inviabilizando uma comparação mais ampla entres as espécies coletadas. Das duas espécies coletadas de Boodleopsis foram obtidas apenas duas sequências parciais de rbcL para B. vaucherioidea e nenhuma para B. pusilla. A comparação com a única sequência de Boodleopsis depositada nos bancos de dados, uma sequência parcial de rbcL de B. pusilla, revelou baixa divergência molecular com as nossas sequências de B. vaucherioidea. Além da necessidade de obtenção de sequências completas de rbcL de ambas espécies da área estudada para comparação, uma maior amostragem e o emprego de outros marcadores moleculares são necessários para esclarecer o posicionamento taxonômicos desses dois táxons, cuja coespecificidade não pode ser descartada. Morfologicamente, \"Cladophoropsis membranacea\" é uma espécie facilmente identificada, entretanto sequências de ITS obtidas neste estudo são não comparáveis a nenhuma sequência dessa espécie depositada nos bancos de dados, incluindo sequências da localidade tipo. Reconhecidamente Chadophoropsis é um gênero polifilético e integra o complexo Boodlea que inclui diferentes gêneros de Boodleaceae. A obtenção de sequências de outros marcadores como o SSU rDNA e LSU rDNA assim como uma maior amostragem podem ser informativas para esclarecer a posição das \"C. membranacea\" brasileiras dentro das Cladophorales. Mesmo após inúmeras tentativas não foi possível obter sequências para as duas espécies de Rhizoclonium encontradas, R. africanum e R. riparium, cuja identificação foi feita com base em caracteres morfológicos tradicionais / Studies on the diversity of macroalgae from mangroves in Brazil have been based only on morphological approaches, in which characters used are unstable and uninformative for the species identification and delimitation. In this context, macroalgae of mangroves were investigated for the first time in the Brazilian coast using a molecular approach, having as target of our study the mangrove of the Barnabé Island, Santos, São Paulo. DNA Barcode markers UPA and COI-5P were used, besides the rbcL, largely used for phylogenetic inferences, and also SSU rDNA. In addition to these, the ITS and tufA markers were used exclusively for the green algae, the latter unsuccessfully. Fifteen species were recorded for the studied area, ten Rhodophyta and five Chlorophyta. Of these, two are new records for the State of São Paulo, Caloglossa apomeiotica e Boodleopsis vaucherioidea. Of the four species of the genus Bostrychia identified in this study: \"Bostrychia calliptera\", B. montagnei, B. moritziana and B. radicans, only B. montagnei proved to be a molecular and morphologically well-defined species. The other species formed complexes with different molecular lineages. For B. radicans and B. moritziana, the analyses showed three and two molecular lineages, respectively, of the seven identified for the B. radicans/B. moritziana complex in the literature. The taxon identified as \"B. calliptera\" showed high molecular divergence with sequences of B. calliptera from Brazil, presenting morphology \"B. pinnata\", a taxon currently reduced to a synonym of B. calliptera. Caloglossa species, except C. ogasawaraensis, are difficult to identify due to the subtle morphological characters considered of diagnostic value, and their considerable phenotypic plasticity. Caloglossa apomeiotica, C. confusa and C. leprieurii were identified primarily by the use of molecular markers. Caloglossa apomeiotica can be segregated from the others by the presence of bisporangia, which makes unreliable morphological identification when collected infertile thalli, while C. confusa has strongly constricted thallus nodes. The molecular data obtained for Catenella caespitosa from SSU sequences suggest that the citations of this species for the Brazilian coast may be misleading since they have high intraspecific divergence with C. caespitosa from database. Due to the lack of sequences from the type locality, sequences of DNA barcode markers, and a major molecular sampling of Catenella species in databases, became impossible to reach a conclusive result. The obtaining of sequences for green algae was extremely problematic, making impracticable a broader comparison between the collected species. Of the two collected species of Boodleopsis, only two partial sequences of rbcL were obtained for B. vaucherioidea and none for B. pusilla. The comparison with the unique sequence of Boodleopsis deposited in databases, a partial rbcL sequence of B. pusilla, revealed low molecular divergence with our sequences of B. vaucherioidea. Besides the need to obtain complete sequences of rbcL from both species from the studied area for comparison, an increased sampling and the use of other molecular markers are needed to clarify the taxonomic position of these two taxa, whose conspecificity cannot be disregarded. Morphologically, \"Cladophoropsis membranacea\" is an easily identified species, however, ITS sequences obtained in this study are not comparable to any sequence of this species deposited in databases, including sequences from the type locality. Admittedly. Chadophoropsis is a polyphyletic genus and integrates the Boodlea complex that includes different genera of Boodleaceae. The obtaining of sequences from other markers, such as SSU rDNA and LSU rDNA, well as a larger sampling, may be informative to clarify the taxonomic position of \"C. membranacea\" within the Brazilian Cladophorales. Even after numerous attempts we could not get sequences for the two Rhizoclonium species found in the studied area: R. africanum and R. riparium, whose identification was made based on traditional morphological characters

Sympatry is characterized by the existence of two or more species phylogenetically related occupying the same ecological niche. This event has already been recorded in species of crabs of the genus Aegla in Brazil between two species, and though morphological characteristics only. For Aegla, as in most crustaceans, there are few studies to verify if there is any relationship between sympatry and genetic diversity of populations living in this association. In the present study, besides a new record of sympatry between two species of Aegla for Brazil, Aegla platensis and Aegla spinipalma, occurring in Batú River, the occurrence of sympatry among three species, A. platensis, Aegla sp. (in preparation) and Aegla grisella, living in Cambará River, is also presented. To verify the existence of sympatry at the sampling points, besides the morphological characteristics used for the identification of individuals, the technique of DNA-Barcoding was applied using a fragment of 207 bp of the gene COI amplified in 22 individuals and from these sequences interspecific, intra- and interpopulation p-distance values were obtained. Sympatry between two species in Batú River was verified by both morphological and molecular data. For the species living in Cambará River mitochondrial and morphological data were not completely congruent, because some sequences of A. grisella clustered with individuals of other species, presenting low values of interspecific distance. However, these sequences probably are nuclear pseudogenes and when they were removed from the analysis, three species were identified in Cambará River. To verify if there is any relationship between sympatry and genetic diversity of A. platensis, two populations living in sympatry (Cambará and Batú rivers) and one allopatric population from Fiuza River were used. The technique of AFLP was applied to 120 individuals (20 from each population) revealing a high genetic variability for all populations and no relationship between sympatry and level of genetic diversity was found in A. platensis. A high population structuring was observed among populations, which is probably due to the distance among populations and to the low dispersion capability in aeglids. Results from the COI mitochondrial data and from the AFLP nuclear data presented some incongruence when compared, since using AFLP all the A. platensis populations and also the other species were separated through Bayesian analysis and UPGMA, but the same was not found for COI data. The incongruence between mitochondrial and nuclear data reinforces the idea that some A. grisella sequences are in fact nuclear pseudogenes co-amplified in PCR, due to the utilization of degenerated universal primers. / Simpatria é caracterizada pela existência de duas ou mais espécies filogeneticamente próximas ocupando um mesmo nicho ecológico. Este evento já foi registrado em espécies de caranguejos do gênero Aegla no Brasil, sendo estes registros realizados apenas entre duas espécies e utilizando para isto, apenas características morfológicas. Para Aegla, assim como na maioria dos crustáceos, existem poucos estudos destinados a avaliar se há alguma relação entre a simpatria e a diversidade genética das populações que vivem em tal associação. Neste estudo, além de ser apresentado mais um registro de simpatria entre duas espécies de eglídeos para o Brasil, sendo estas Aegla platensis e Aegla spinipalma ocorrendo no Rio Batú, também é apresentada ocorrência de simpatria entre três espécies, sendo estas A. platensis, Aegla sp. (em preparação) e Aegla grisella ocorrendo no Rio Cambará. Para verificar a existência de simpatria nos pontos de amostragem, além das características morfológicas utilizadas para a identificação dos indivíduos, foi realizada a técnica de DNA Barcoding com um fragmento de 207 pb do gene COI, amplificado em 22 indivíduos e a partir dessas sequências foram obtidos os valores de distâncias interespecífica, intra e interpopulacional através do modelo de distância p. A constatação da existência de simpatria entre duas espécies no Rio Batú foi verificada tanto pelos dados morfológicos quanto para os dados moleculares, porém para as espécies do rio Cambará os dados mitocondriais não foram totalmente congruentes com os morfológicos, pois algumas sequências de A. grisella agruparam com sequências de indivíduos de outras espécies e apresentaram baixos valores de distância interespecífica. Entretanto, estas sequências provavelmente são pseudo-genes nucleares e quando retiradas das análises permitiram a identificação de três espécies simpátricas no Rio Cambará. A fim de avaliar se existe alguma relação entre a simpatria e a diversidade genética de A. platensis foram utilizadas duas populações simpátricas desta espécie (Rios Cambará e Batú) e uma população alopátrica, oriunda do Rio Fiúza. A técnica de AFLP foi utilizada em 120 indivíduos (20 de cada população), onde constatou-se a existência de uma alta variabilidade genética em todas as populações, não tendo sido encontrada relação entre a simpatria e o nível de diversidade genética em A. platensis. Uma alta estruturação genética observada entre as populações, que provavelmente está associada à distância entre elas e à baixa capacidade de dispersão dos eglídeos. Os resultados do marcador mitocondrial COI e do marcador nuclear AFLP apresentaram algumas incongruências, quando comparados, visto que usando AFLP todas as populações de A. platensis e das demais espécies foram separadas por meio de análise Bayesiana e UPGMA, o mesmo não ocorrendo para o COI. A incongruência entre os dados nucleares e mitocondriais reforça a ideia de que algumas sequências de A. grisella são na verdade pseudo-genes nucleares coamplificados com o gene COI, devido à utilização de primers universais degenerados na PCR.

The thesis deals with the hemiparasitic species Melampyrum pratense. This species was monitored during the growing season 2009 and the relationship between morphological characteristics and reproductive success of plant individuals was assessed. The relationships between population density, the vegetative biomass and the investments in generative reproduction were investigated. Identity of the host species was determined using the DNA barcoding method.

Estudos sobre a diversidade de macroalgas de manguezais no Brasil tem-se baseado apenas em abordagens morfológicas, nas quais os caracteres empregados são instáveis e pouco informativos para a identificação e delimitação de espécies. Neste contexto, macroalgas de manguezais foram investigadas pela primeira vez no litoral brasileiro usando uma abordagem molecular, tendo como alvo de estudo o manguezal da Ilha Barnabé, Baixada Santista, São Paulo. Foram utilizados marcadores moleculares do tipo Barcode, UPA e COI-5P, além do rbcL, amplamente empregado para inferências filogenéticas, e o SSU rDNA. Além desses, os marcadores ITS e tufA foram empregados exclusivamente para as algas verdes, este último sem sucesso. Quinze espécies foram registradas para a área estudada, sendo dez Rhodophyta e cinco Chlorophyta. Destas, duas não novas ocorrências para o Estado de São Paulo, Caloglossa apomeiotica e Boodleopsis vaucherioidea. Das quatro espécies do gênero Bostrychia identificadas neste estudo: \"Bostrychia calliptera\", B. montagnei, B. moritziana e B. radicans, apenas B. montagnei revelou-se uma espécie molecular e morfologicamente bem definida. As demais formaram complexos de espécies com linhagens moleculares distintas. Para B. radicans e B. moritziana as análises relevaram três e duas linhagens moleculares, respectivamente, das sete identificadas para o complexo B. radicans/B. moritziana na literatura. O táxon identificado como \"B. calliptera\" mostrou alta divergência molecular com sequências de B. calliptera do Brasil, apresentando morfologia \"B. pinnata\", um táxon atualmente reduzido a sinônimo de B. calliptera. Espécies do gênero Caloglossa, excetuando C. ogasawaraensis, são de difícil identificação morfológica devido aos tênues caracteres considerados de valor diagnóstico e a sua considerável plasticidade fenotípica. Caloglossa apomeiotica, C. confusa e C. leprieurii foram identificadas essencialmente com o emprego de marcadores moleculares. Caloglossa apomeiotica pode ser segregada das demais pela presença de biesporângios, o que impossibilita uma identificação morfológica segura quando coletados talos inférteis, enquanto C. confusa possui nós fortemente contritos. Os dados moleculares obtidos para Catenella caespitosa a partir de sequências de SSU sugerem que as citações dessa espécie para o litoral brasileiro podem estar equivocadas já que apresentam alta divergência intraespecífica com C. caespitosa do banco de dados. A falta de sequências da localidade tipo, de sequências com marcadores do tipo Barcode e de uma maior amostragem molecular das espécies de Catenella nos bancos de dados, nos impossibilitaram chegar a um resultado conclusivo. A obtenção de sequências para as algas verdes foi extremamente problemática, inviabilizando uma comparação mais ampla entres as espécies coletadas. Das duas espécies coletadas de Boodleopsis foram obtidas apenas duas sequências parciais de rbcL para B. vaucherioidea e nenhuma para B. pusilla. A comparação com a única sequência de Boodleopsis depositada nos bancos de dados, uma sequência parcial de rbcL de B. pusilla, revelou baixa divergência molecular com as nossas sequências de B. vaucherioidea. Além da necessidade de obtenção de sequências completas de rbcL de ambas espécies da área estudada para comparação, uma maior amostragem e o emprego de outros marcadores moleculares são necessários para esclarecer o posicionamento taxonômicos desses dois táxons, cuja coespecificidade não pode ser descartada. Morfologicamente, \"Cladophoropsis membranacea\" é uma espécie facilmente identificada, entretanto sequências de ITS obtidas neste estudo são não comparáveis a nenhuma sequência dessa espécie depositada nos bancos de dados, incluindo sequências da localidade tipo. Reconhecidamente Chadophoropsis é um gênero polifilético e integra o complexo Boodlea que inclui diferentes gêneros de Boodleaceae. A obtenção de sequências de outros marcadores como o SSU rDNA e LSU rDNA assim como uma maior amostragem podem ser informativas para esclarecer a posição das \"C. membranacea\" brasileiras dentro das Cladophorales. Mesmo após inúmeras tentativas não foi possível obter sequências para as duas espécies de Rhizoclonium encontradas, R. africanum e R. riparium, cuja identificação foi feita com base em caracteres morfológicos tradicionais / Studies on the diversity of macroalgae from mangroves in Brazil have been based only on morphological approaches, in which characters used are unstable and uninformative for the species identification and delimitation. In this context, macroalgae of mangroves were investigated for the first time in the Brazilian coast using a molecular approach, having as target of our study the mangrove of the Barnabé Island, Santos, São Paulo. DNA Barcode markers UPA and COI-5P were used, besides the rbcL, largely used for phylogenetic inferences, and also SSU rDNA. In addition to these, the ITS and tufA markers were used exclusively for the green algae, the latter unsuccessfully. Fifteen species were recorded for the studied area, ten Rhodophyta and five Chlorophyta. Of these, two are new records for the State of São Paulo, Caloglossa apomeiotica e Boodleopsis vaucherioidea. Of the four species of the genus Bostrychia identified in this study: \"Bostrychia calliptera\", B. montagnei, B. moritziana and B. radicans, only B. montagnei proved to be a molecular and morphologically well-defined species. The other species formed complexes with different molecular lineages. For B. radicans and B. moritziana, the analyses showed three and two molecular lineages, respectively, of the seven identified for the B. radicans/B. moritziana complex in the literature. The taxon identified as \"B. calliptera\" showed high molecular divergence with sequences of B. calliptera from Brazil, presenting morphology \"B. pinnata\", a taxon currently reduced to a synonym of B. calliptera. Caloglossa species, except C. ogasawaraensis, are difficult to identify due to the subtle morphological characters considered of diagnostic value, and their considerable phenotypic plasticity. Caloglossa apomeiotica, C. confusa and C. leprieurii were identified primarily by the use of molecular markers. Caloglossa apomeiotica can be segregated from the others by the presence of bisporangia, which makes unreliable morphological identification when collected infertile thalli, while C. confusa has strongly constricted thallus nodes. The molecular data obtained for Catenella caespitosa from SSU sequences suggest that the citations of this species for the Brazilian coast may be misleading since they have high intraspecific divergence with C. caespitosa from database. Due to the lack of sequences from the type locality, sequences of DNA barcode markers, and a major molecular sampling of Catenella species in databases, became impossible to reach a conclusive result. The obtaining of sequences for green algae was extremely problematic, making impracticable a broader comparison between the collected species. Of the two collected species of Boodleopsis, only two partial sequences of rbcL were obtained for B. vaucherioidea and none for B. pusilla. The comparison with the unique sequence of Boodleopsis deposited in databases, a partial rbcL sequence of B. pusilla, revealed low molecular divergence with our sequences of B. vaucherioidea. Besides the need to obtain complete sequences of rbcL from both species from the studied area for comparison, an increased sampling and the use of other molecular markers are needed to clarify the taxonomic position of these two taxa, whose conspecificity cannot be disregarded. Morphologically, \"Cladophoropsis membranacea\" is an easily identified species, however, ITS sequences obtained in this study are not comparable to any sequence of this species deposited in databases, including sequences from the type locality. Admittedly. Chadophoropsis is a polyphyletic genus and integrates the Boodlea complex that includes different genera of Boodleaceae. The obtaining of sequences from other markers, such as SSU rDNA and LSU rDNA, well as a larger sampling, may be informative to clarify the taxonomic position of \"C. membranacea\" within the Brazilian Cladophorales. Even after numerous attempts we could not get sequences for the two Rhizoclonium species found in the studied area: R. africanum and R. riparium, whose identification was made based on traditional morphological characters

Lithophylloideae é uma subfamília de algas calcárias que inclui tanto gêneros articulados quanto os não articulados, que por sua vez compreende gêneros incrustantes e um moniliforme. Pela primeira vez a diversidade específica de Amphiroa, Lithophyllum, Paulsilvella e Titanoderma para o Brasil é investigada de maneira somatória e comparativa, quanto a uma variedade de aspectos que incluíram os morfológicos/anatômicos, ecológicos e moleculares (incluindo a própria avaliação das ferramentas moleculares e das características taxonomicamente importantes). Encontramos 26 espécies, sendo quinze Amphiroa, oito Lithophyllum, uma Paulsilvella e duas Titanoderma: Amphiroa rigida, A. vanbosseae, Amphiroa sp. 1, Amphiroa sp. 2, Amphiroa sp. 3, Amphiroa sp. 4, seis espécies para o Complexo A. fragilíssima, três espécies para o Complexo A. beauvoisii, Lithophyllum atlanticum, L. corallinae, Lithophyllum cf. depressum, Lithophyllum sp. 1, Lithophyllum sp. 2, Lithophyllum sp. 3, Lithophyllum sp. 4, Lithophyllum cf. margaritae, Paulsilvella huveorum, Titanoderma pustulatum e T. Prototypum. Para a caracterização molecular utilizamos quatro marcadores principais: UPA, COI-5P, psbA e rbcL-3P, atendendo a dois objetivos: a delimitação de espécies e a avaliação destes diferentes marcadores como DNA Barcodes para as espécies de Lithophylloideae dentro do contexto da variabilidade encontrada na costa brasileira. Para isto utilizamos cinco métodos de agrupamento: Taxas de Divergência, ABGD, MDS, PTP e SPN. A decisão final quanto à quantidade de espécies presentes é um consenso dos resultados encontrados para todos os marcadores e para todos os métodos de análise. Os casos conflitantes também foram observados quanto aos valores de divergência e de barcoding gap. Os quatro marcadores testados são confiáveis para a delimitação de espécies em Lithophylloideae, proporcionando ótimos valores de suporte para as espécies e valores significativos de barcoding gap. Apesar disto, dada a possibilidade de usar o rbcL-3P também para para fins filogenéticos, ressaltamos a aplicabilidade deste marcador, sendo o indicado caso exista a necessidade de se indicar um único marcador como DNA Barcode para este grupo de algas. A inferência das relações filogenéticas dentro de Lithophylloideae inclui os primeiros dados moleculares para Paulsilvella, através do psbA, do rbcL -3P e também do SSU rDNA. As análises indicam que Amphiroa é monofilético, enquanto que Lithophyllum e Titanoderma provavelmente são polifiléticos. Adicionalmente, é possível que Paulsilvella represente a primeira forma morfológica da subfamília. A distribuição de Amphiroa, Lithophyllum e Titanoderma ao longo da costa brasileira embasou a discussão quanto à distribuição das macroalgas marinhas bentônicas na costa, adicionando ao cenário o papel das correntes costeiras e do Giro do Atlântico Sul. Os dados moleculares possibilitaram ainda a observação do panorama de ocorrência das 26 espécies de Lithophylloideae quanto a sua ocorrência na coluna d\'água (profundidade). Para Amphiroa a maior diversidade está no mesolitoral, enquanto que para Lithophyllum e Titanoderma a maior diversidade se encontra no infralitoral. Oito espécies são restritas ao mesolitoral, enquanto que dez (incluindo P. huveorum) são restritas ao infralitoral. Estes fatos chamam a atenção da necessidade de se incluir diferentes técnicas de coleta nos trabalhos de diversidade em Florideophycideae, do contrário podemos negligenciar uma diversidade inexplorada / Lithophylloideae is a subfamily within Corallinaceae including both articulated and non-articulated genera. This work is the first effort to access the specific diversity for Amphiroa, Lithophyllum, Paulsilvella and Titanoderma for Brazil based on a sum of morphological anatomical, ecological and molecular data, including also the tools applicability evaluation. From the 26 species found, are Amphiroa, eight are Lithophyllum, two are Titanoderma and one is the first register for Paulsilvella for Atlantic waters: Amphiroa rigida, A. vanbosseae, Amphiroa sp. 1, Amphiroa sp. 2, Amphiroa sp. 3, Amphiroa sp. 4, six criptic species for the A. fragilissima Complex, three cryptic species for the A. beauvoisii Complex, Lithophyllum atlanticum, L. corallinae, Lithophyllum cf. depressum, Lithophyllum sp. 1, Lithophyllum sp. 2, Lithophyllum sp. 3, Lithophyllum sp. 4, Lithophyllum cf. margaritae, Titanoderma pustulatum, T. prototypum and Paulsilvella huveorum. For the molecular characterization four markers were used: UPA, COI-5P, psbA and rbcL-3P, which have attended to two the delimitation of species and also to the evaluation of these markers as DNA Barcodes for the species of Lithophylloideae within the context of variability found on the Brazilian coast. For these two purposes we have applied five clustering methods, based on the Divergence Rates, ABGD, MDS, PTP and SPN. Conflicting cases were also observed for values of barcoding gap. The four molecular markers are reliable for species delimitation within the Lithophylloideae, providing good support values for the species and significant amounts of barcoding gap. Nevertheless, given the possibility of using the rbcL marker also for phylogenetic purposes, we emphasize the applicability of rbcL-3P as the most appropriate marker, in case there is a need to indicate a single marker as DNA Barcode for this group alga. A phylogenetic inference for the Lithophylloideae included the first sequences for Paulsilvella and used psbA, rbcL-3P and SSU rDNA. We found that Amphiroa is monophyletic while Lithophyllum and Titanoderma are probably polyphyletic. Additionally, Paulsilvella may represent the first morphological form of the subfamily. Amphiroa, Lithophyllum and Titanoderma distribution along the Brazilian coast have allowed a new discussion concerning the benthic macroalgae distribution to the country, adding two new factors to this scenario: the coastal currents movement and the South Atlantic Gyre. The molecular data have also allowed accessing the preferences according to the depth on the water column for the 26 species. The major diversity for Amphiroa grows at the intertidal, while Lithophyllum and Titanoderma diversity is higher at the subtidal and Paulsilvella is restricted to the subtidal. There are eight species restricted to the intertidal and ten restricted to the subtidal (including P. huveorum), what addresses to a need to include these both environments on diversity studies for the Florideophycideae otherwise it will imply the existence of an unexplored diversity

As algas da ordem Halymeniales (Rhodophyta) apresentam grande importância ecológica e econômica como produtores primários e de compostos bioativos, além de incluírem espécies invasoras em várias localidades do mundo. A taxonomia do grupo é bastante problemática, com vários registros de identificações equivocadas e mudanças nomenclaturais. Em virtude disso, diversos estudos têm incluído ferramentas moleculares como auxílio à taxonomia morfológica do grupo. O objetivo deste estudo é investigar por meio de técnicas moleculares e morfológicas a diversidade da ordem Halymeniales no litoral do Brasil de forma, a contribuir para o conhecimento da flora marinha do país. Para isso, foram sequenciados três marcadores moleculares, UPA, COI-5P e rbcL, cujos dados, combinados com a observação de caracteres morfológicos, resultaram na delimitação de 26 espécies. Dessas, 11 são espécies novas para a ciência: Corynomorpha cf. clavata, dois táxons morfologicamente identificados como \"Cryptonemia\" crenulata, dois táxons morfologicamente identificados como Cryptonemia seminervis, Halymenia ignifera, Halymenia pinnatifida, Halymenia silviae, Grateloupia cf. dichotoma, Grateloupia cf. filicina 1 e Grateloupia cf. filicina 2. Pelo menos sete gêneros novos foram encontrados, representados pelos táxons: \"Cryptonemia\" bengryi, integrantes do complexo \"Cryptonemia\" crenulata, \"Cryptonemia\" delicatula, \"Halymenia\" elongata, \"Halymenia\" floridana, Halymeniales sp. 1 e Halymeniales sp. 2. Foram encontradas três espécies cuja localidade-tipo é a Ásia: Grateloupia orientalis, Grateloupia turuturu e Grateloupia yangjiangensis. Entre os táxons previamente citados para a costa brasileira, nove não foram encontrados. Se os táxons não encontrados constituírem espécies válidas, a diversidade da ordem no Brasil será de 35 espécies. As análises filogenéticas mostraram que os gêneros Cryptonemia, Halymenia e Grateloupia constituem grupos não-monofiléticos. Os resultados demonstraram a existência de ampla diversidade críptica e pseudo-críptica e de espécies e gêneros novos para a ciência, além de revelarem a ocorrência de espécies não nativas. Este estudo contribui substancialmente para o conhecimento da diversidade de algas marinhas na costa brasileira / Algae of Halymeniales (Rhodophyta) present a wide ecological and economic importance, as primary producers and bioactive compounds producers, and include invasive species worldwide. Its taxonomy is quite problematic, with reports of misidentifications and nomenclatural changes. Therefore, studies have included molecular tools to assist the morphological taxonomy of this order. This study aims to investigate through molecular and morphological techniques the diversity of Halymeniales along the Brazilian coast, in order to contribute to the knowledge of native marine flora. Three molecular markers were sequenced, UPA, COI-5P and rbcL, whose data were allied to morphological characters and resulted in 26 delimited species. There are 11 new species to science: Corynomorpha cf. clavata, two taxa morphologically identified as \"Cryptonemia\" crenulata, two taxa morphologically identified as Cryptonemia seminervis, Halymenia ignifera, Halymenia pinnatifida, Halymenia silviae, Grateloupia cf. dichotoma, Grateloupia cf. filicina 1 and Grateloupia cf. filicina 2. At least seven new genera were found, represented by the following taxa: \"Cryptonemia\" bengryi, representatives of \"Cryptonemia\" crenulata complex, \"Cryptonemia\" delicatula, \"Halymenia\" elongata, \"Halymenia\" floridana, Halymeniales sp. 1 and Halymeniales sp. 2. Three species whose type locality is Asia were detected: Grateloupia orientalis, Grateloupia turuturu and Grateloupia yangjiangensis. Among taxa previously recorded to Brazilian coast, nine were not found. If those taxa constitute valid species, the diversity of the order in Brazil will be represented by 35 species. Phylogenetic analyses showed that Cryptonemia, Halymenia and Grateloupia constitute non-monophyletic groups. Results demonstrated the existence of wide cryptic and pseudo-cryptic diversity as well as novel species and genera, and revealed the presence of non-native species

Taxonomie - die Determination von Arten - ermöglicht ein grundlegendes Verständnis ökologischer Zusammenhänge und der Evolution. Heutzutage ist sie aufgrund innovativer Forschung, evolutiver Fragestellungen und politisch relevanter Ziele zur Erhaltung der globalen Biodiversität in viele Disziplinen eingebunden (LOHRMANN et al. 2012). „Beispiele sind die Identifikation invasiver Arten, die nachhaltige Bewirtschaftung mariner Ressourcen oder die Entwicklung neuer Medikamente. Zudem ist die Kenntnis der Arten und ihrer Interaktionen nötig, um im politischen Raum die Erreichung - oder auch Nichteinhaltung - von Biodiversitäts- und Nachhaltigkeitszielen zu dokumentieren“ (LOHRMANN et al. 2012). Daher sind aussagekräftige Monitorings und Modelle zur Entwicklung unserer Ökosysteme nur mit der nötigen Artenkenntnis und mit Hilfe taxonomischer Methoden möglich.

The ecotoxicological literature reveals that countless researchers worldwide rely upon informally identified commercial earthworm stocks for laboratory bioassays. The primary aim of this study was to investigate laboratory and commercial stocks of Eisenia species used in South Africa in order to confirm their taxonomy, assess their levels of genetic richness and differentiation. To do so, populations of potential Eisenia andrei and Eisenia fetida were purchased/obtained from vermiculturists and laboratories from four provinces of South Africa. DNA barcoding was used to investigate these taxonomic uncertainties. The COI gene was partially amplified and sequenced in selected earthworms from eight local populations (focal groups) and two European laboratory stocks (non-focal groups). Only nine COI haplotypes were identified from the 224 sequences generated. One of these haplotypes was found to belong to the Megascolecidae Perionyx excavatus. The remaining eight haplotypes belonged to the genus Eisenia although only a single Eisenia fetida haplotype, represented by six specimens, was found in one of the European populations. The other seven haplotypes, all occurring in South Africa, were Eisenia andrei. No Eisenia fetida was found in the South African based populations. One of the commercial stocks from South Africa and a laboratory culture from Europe were mixes of E. andrei - P. excavatus and E. andrei – E. fetida respectively. COI haplotype numbers were limited to two to three distinct sequences within each of the local groups. This translated into a haplotype diversity (H) lower than 0.45 in all the populations, which is very low when compared to other such earthworm studies in which COI polymorphism has been investigated. Of all the local populations investigated, only the lone field population included was genetically divergent from the other populations. This was explained by the haplotype distribution across the populations which indicated that this population was the only one not harbouring the haplotype which represented 75% or more of the COI sequences within the local populations. Because research suggests that earthworm populations with limited genetic diversity may suffer inbreeding depression which could affect traits such as reproduction and survival, the secondary aim was to test whether metal-sensitive earthworms were overly present in the populations investigated. To do so, the three most common COI haplotypes identified between the 8 local populations of E. andrei (called Hap1, Hap2 and Hap3) were paired up and exposed to cadmium. A total of six couples were exposed to 0, 25, 50 and 100 mg Cd/kg for 4 weeks at 20ºC. The survival, biomass variation, cocoon production and cocoon hatching success were assessed for all the couples. The results indicated that couple 6 (Hap3xHap3) was the most sensitive for three of the endpoints assessed whereas couple 4 (Hap1 x Hap3) was the least sensitive. Cocoon hatching success could not help differentiate the couples. The analysis of Cd tissue contents revealed that with increasing Cd concentration, Cp6 (Hap3xHap3) could accumulate significantly more Cd than any other couple (p ≤ 0.01). These findings indicate that earthworm populations may carry intrinsically metal-tolerant and metal-sensitive genotypes. In the context of ecotoxicological testing, the present results underline the importance of using genetically diverse populations in laboratory testing as Cp6 (Hap3xHap3) could have suffered from the deleterious effect of inbreeding. Because E. fetida could not be found in the local populations assessed, it is recommended that further earthworm DNA barcoding studies, covering a more representative geographical area of South Africa and including more field populations of Eisenia spp. be conducted. Because of the occurrence of genetic homogeneity in the populations studied, it is suggested that captive breeding initiatives be established using specimens obtained from several geographically distant field and reared populations. Further research investigating patterns of Cd accumulation/excretion kinetics between the Cd-tolerant and Cd-sensitive individuals reported in the present study, should be conducted to help determine whether inbreeding is the sole factor explaining the observed genotypic responses to Cd. Finally, the necessity of a standardised earthworm barcoding protocol that could help both to properly identify laboratory earthworm stocks and to select genetically diverse stocks suitable for laboratory testing, is discussed together with the relevance of the present work to ecotoxicological testing in general. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

The ecotoxicological literature reveals that countless researchers worldwide rely upon informally identified commercial earthworm stocks for laboratory bioassays. The primary aim of this study was to investigate laboratory and commercial stocks of Eisenia species used in South Africa in order to confirm their taxonomy, assess their levels of genetic richness and differentiation. To do so, populations of potential Eisenia andrei and Eisenia fetida were purchased/obtained from vermiculturists and laboratories from four provinces of South Africa. DNA barcoding was used to investigate these taxonomic uncertainties. The COI gene was partially amplified and sequenced in selected earthworms from eight local populations (focal groups) and two European laboratory stocks (non-focal groups). Only nine COI haplotypes were identified from the 224 sequences generated. One of these haplotypes was found to belong to the Megascolecidae Perionyx excavatus. The remaining eight haplotypes belonged to the genus Eisenia although only a single Eisenia fetida haplotype, represented by six specimens, was found in one of the European populations. The other seven haplotypes, all occurring in South Africa, were Eisenia andrei. No Eisenia fetida was found in the South African based populations. One of the commercial stocks from South Africa and a laboratory culture from Europe were mixes of E. andrei - P. excavatus and E. andrei – E. fetida respectively. COI haplotype numbers were limited to two to three distinct sequences within each of the local groups. This translated into a haplotype diversity (H) lower than 0.45 in all the populations, which is very low when compared to other such earthworm studies in which COI polymorphism has been investigated. Of all the local populations investigated, only the lone field population included was genetically divergent from the other populations. This was explained by the haplotype distribution across the populations which indicated that this population was the only one not harbouring the haplotype which represented 75% or more of the COI sequences within the local populations. Because research suggests that earthworm populations with limited genetic diversity may suffer inbreeding depression which could affect traits such as reproduction and survival, the secondary aim was to test whether metal-sensitive earthworms were overly present in the populations investigated. To do so, the three most common COI haplotypes identified between the 8 local populations of E. andrei (called Hap1, Hap2 and Hap3) were paired up and exposed to cadmium. A total of six couples were exposed to 0, 25, 50 and 100 mg Cd/kg for 4 weeks at 20ºC. The survival, biomass variation, cocoon production and cocoon hatching success were assessed for all the couples. The results indicated that couple 6 (Hap3xHap3) was the most sensitive for three of the endpoints assessed whereas couple 4 (Hap1 x Hap3) was the least sensitive. Cocoon hatching success could not help differentiate the couples. The analysis of Cd tissue contents revealed that with increasing Cd concentration, Cp6 (Hap3xHap3) could accumulate significantly more Cd than any other couple (p ≤ 0.01). These findings indicate that earthworm populations may carry intrinsically metal-tolerant and metal-sensitive genotypes. In the context of ecotoxicological testing, the present results underline the importance of using genetically diverse populations in laboratory testing as Cp6 (Hap3xHap3) could have suffered from the deleterious effect of inbreeding. Because E. fetida could not be found in the local populations assessed, it is recommended that further earthworm DNA barcoding studies, covering a more representative geographical area of South Africa and including more field populations of Eisenia spp. be conducted. Because of the occurrence of genetic homogeneity in the populations studied, it is suggested that captive breeding initiatives be established using specimens obtained from several geographically distant field and reared populations. Further research investigating patterns of Cd accumulation/excretion kinetics between the Cd-tolerant and Cd-sensitive individuals reported in the present study, should be conducted to help determine whether inbreeding is the sole factor explaining the observed genotypic responses to Cd. Finally, the necessity of a standardised earthworm barcoding protocol that could help both to properly identify laboratory earthworm stocks and to select genetically diverse stocks suitable for laboratory testing, is discussed together with the relevance of the present work to ecotoxicological testing in general. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Thesis (PhD (Food Sc)--Stellenbosch University, 2011. / ENGLISH ABSTRACT: Consumers have the right to accurate information on the fish products they purchase to enable them to make educated seafood selections that will not endanger their own wellbeing or the wellbeing of the environment. Unfortunately, marine resource scarcity, financial incentives and inadequate or poorly enforced regulations have all promoted the mislabelling of fish species on global markets, the results of which may hold economic, conservation and health consequences. The primary aims of this study were to determine the most commonly available fish species on the South African market, to establish and compare DNA-based methods for the unambiguous identification of these species and to utilise the most applicable methods to evaluate the extent of mislabelling on the local fisheries market. The results from surveys of n = 215 restaurants and n = 200 retail outlets in four South African provinces (Western Cape, Kwa-Zulu Natal, Eastern Cape and Gauteng) indicated that 34 and 70 nominal fish types were available in restaurants and retail outlets, respectively, the most common of which were kingklip, salmon and hake. Over 30% of the fish species being sold were of conservation concern, while several outlets marketed specially-protected, illegal-to-sell species in South Africa. Fish purveyors were poorly equipped to provide information on the identity, origin, production method (farmed/wild) and sustainability of the fish they were selling and the labelling of many packaged fish products was in contravention with South African regulations. Data were published for the first time comparing the efficiency of five methods (urea-SDS-proteinase K, phenol-chloroform, salt extraction, SureFood PREP kit and Wizard Genomic DNA Purification kit) for the extraction of DNA from the muscle tissue of fish species available in South Africa. The SureFood kit was identified as the most suitable method for DNA extraction from fish muscle, extracting significantly (P < 0.05) higher DNA yields than all other methods evaluated and being simple and safe to use. A comprehensive reference library of genetic information was compiled for the first time that contains sufficient DNA sequence data from different mitochondrial DNA loci (16S ribosomal RNA (rRNA), 12S rRNA and cytochrome c oxidase I (COI) genes, as well as the control region) to allow the explicit identification of 53 fish species in South Africa. Although 16S and 12S rRNA gene sequencing allowed the identification of most fish to the genus level, the discrimination of closely-related, congeneric species was problematic when based on these gene regions. Conversely, the vast majority (98%) of fish examined could be readily differentiated by their COI sequences, with only members of the genus Thunnus requiring supplementary control region sequencing for species confirmation. Lastly, sequencing of the COI region was used to show that 9% of fish samples collected from local seafood wholesalers and 31% of samples from retail outlets were mislabelled. This study has established that fish mislabelling is a reality on the South African market and that DNA-based methods should be applied by both industry and regulatory bodies to deter illegal activities and to promote transparency on the domestic fisheries market. / AFRIKAANSE OPSOMMING: Verbruikers het die reg tot akkurate informasie rakende die visprodukte wat hulle aankoop. Hierdie inligting sal hulle bemagtig om ingeligte seekos keuses te maak wat voordelig sal wees vir beide die verbruiker se eie, sowel as die omgewing, se voortbestaan. Ongelukkig het 'n gebrek aan seelewebronne, geldelike aansporings en onvanpaste of swak geïmplimenteerde regulasies gelei tot die verkeerde etikettering van visspesies op die wêreldmarkte. Dit mag ekonomiese-, bewarings- en gesondheidsgevolge inhou. Die primêre doelwitte van hierdie studie was om te bepaal watter visspesies die algemeenste beskikbaar is in die Suid-Afrikaanse mark, om DNS-gebaseerde metodes vir die duidelike identifisering van hierdie spesies te vind en te vergelyk, en om die mees gepaste metodes te gebruik om die omvang van verkeerde etikettering in die plaaslike vismarkte te evalueer. Die resultate van opnames van n = 215 restaurante en n = 200 winkels in vier Suid-Afrikaanse provinsies (Wes-Kaap, Kwa-Zulu Natal, Oos-Kaap en Gauteng) het gewys dat 34 en 70 nominale visspesies in onderskeidelik restaurante en kleinhandelaars beskikbaar was. Koningklip, salm en stokvis was die mees algemene spesies. Meer as 30% van die visspesies wat te koop was is van bewaringsbelang, terwyl verskeie winkels spesiaal-beskermde, onwettig-om-te-verkoop spesies in Suid-Afrika bemark het. Visverkopers was swak bemagtig om informasie oor die identiteit, oorsprong, produksiemetode (teel/wild) en volhoubaarheid van die vis wat hulle verkoop het te kon gee. Verder was die etikettering van baie verpakte visprodukte in stryd met Suid-Afrikaanse regulasies. Vir die eerste keer is data gepubliseer wat vyf metodes (ureum-SDS-proteïenase K, fenolchloroform, sout-ekstraksie, SureFood PREP stel en Wizard Genomic DNS suiwering stel) vergelyk in hul doeltreffendheid om DNS vanuit die spierweefsel van visspesies wat in Suid-Afrika beskikbaar is te ekstraheer. Die SureFood stel is as die mees geskikte metode vir DNS ekstraksie vanuit visweefsel geïdentifiseer aangesien die DNS opbrengs betekenisvol (P < 0.05) hoër was met hierdie metode, en dit ook 'n eenvoudige en veilige metode is om te gebruik. 'n Omvattende verwysingsbiblioteek van genetiese informasie wat voldoende DNS volgordebepalingsdata van verskillende mitokondriale DNS lokusse (16S ribosomale RNS (rRNS), 12S rRNS en sitochroom c oksidase I (COI) gene, sowel as die kontrolegebiede) bevat, is vir die eerste keer opgestel om die besliste identifisering van 53 visspesies in Suid-Afrika toe te laat. Alhoewel 16S en 12S rRNS geenvolgordebepaling die identifisering van meeste visse op genusvlak toegelaat het, was die diskriminasie van naby-verwante, gelyksoorting spesies problematies wanneer hierdie geengebiede gebruik is. Die oorgrote meerderheid (98%) vis wat ondersoek is geredelik onderskei op grond van hul COI volgordebepalings, met slegs lede van die genus Thunnus wat addisionele kontrolegebied volgordebepaling vir spesies bevestiging vereis het. Laastens, is volgordebepaling van die COI-gebied gebruik om te wys dat 9% van die vismonsters van plaaslike seekosgroothandelaars en 31% van die monsters van kleinhandelaars verkeerd geëtiketteer is. Hierdie studie het bevestig dat die verkeerde etikettering van vis in Suid-Afrika 'n realiteit is, en dat DNS-gebaseerde metodes gebruik moet word deur die industrie sowel as die regulerende liggame om onwettige aktiwiteite teen te werk en om deursigtigheid in plaaslike vismarkte te bevorder.