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

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

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

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

Vetiver grass or Chrysopogon zizanioides (L.) Roberty (1960) is a sterile grass which can regenerate vegetatively from clumps of the rootstock. This, as well as its vigorous and deep root system and flood tolerance makes it an ideal candidate for the use in soil remediation and erosion control. In South Africa, Hydromulch (Pty) Ltd. is part of the landscape, soil reclamation and erosion control industry. The company uses vetiver grass on a wide scale and has accumulated a collection of isolates to serve as possible germ lines for industrial use. Due to the different approaches in environmental management as well as environmental factors, a variety of ecotypes form during the planting and acclimatisation of this genus. Chrysopogon nigritanus (Benth.) Veldkamp (1999), which is a native species to Africa, is closely related to C. zizanioides and differs only slightly from C. zizanioides on a morphological level. The major difference between the two species is that C. nigritanus is able to seed freely and thus the use of this species should be avoided. The need arose to screen other non-fertile plants to uncover additional genotypic variety to enable diversification of vetiver plantings. The aim of this study was to characterise the genotype of 19 isolates of vetiver obtained from Hydromulch (Pty) Ltd. via sequencing analyses of three DNA fragments, ITS, ndhF and rbcL. In addition, the radial root anatomy was also investigated and compared with the genetic analyses. According to the results generated during this study, very little or no genotypical differences exist amongst the different isolates available from the Hydromulch (Pty) Ltd. plant collection. Only in the case of the ITS inference were differences observed between three of the studied isolates. There was no significant difference between the different isolates based on the root anatomy, with the exception of two of the studied isolates which formed starch granules. / MSc (Botany), North-West University, Potchefstroom Campus, 2014

Vetiver grass or Chrysopogon zizanioides (L.) Roberty (1960) is a sterile grass which can regenerate vegetatively from clumps of the rootstock. This, as well as its vigorous and deep root system and flood tolerance makes it an ideal candidate for the use in soil remediation and erosion control. In South Africa, Hydromulch (Pty) Ltd. is part of the landscape, soil reclamation and erosion control industry. The company uses vetiver grass on a wide scale and has accumulated a collection of isolates to serve as possible germ lines for industrial use. Due to the different approaches in environmental management as well as environmental factors, a variety of ecotypes form during the planting and acclimatisation of this genus. Chrysopogon nigritanus (Benth.) Veldkamp (1999), which is a native species to Africa, is closely related to C. zizanioides and differs only slightly from C. zizanioides on a morphological level. The major difference between the two species is that C. nigritanus is able to seed freely and thus the use of this species should be avoided. The need arose to screen other non-fertile plants to uncover additional genotypic variety to enable diversification of vetiver plantings. The aim of this study was to characterise the genotype of 19 isolates of vetiver obtained from Hydromulch (Pty) Ltd. via sequencing analyses of three DNA fragments, ITS, ndhF and rbcL. In addition, the radial root anatomy was also investigated and compared with the genetic analyses. According to the results generated during this study, very little or no genotypical differences exist amongst the different isolates available from the Hydromulch (Pty) Ltd. plant collection. Only in the case of the ITS inference were differences observed between three of the studied isolates. There was no significant difference between the different isolates based on the root anatomy, with the exception of two of the studied isolates which formed starch granules. / MSc (Botany), North-West University, Potchefstroom Campus, 2014

The Calvin-Benson-Bassham (CBB) pathway is the primary pathway for the entry of inorganic carbon in the biosphere. Autotrophic organisms use this cycle to ultimately convert CO2 into carbohydrates using a key enzyme known as ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO). The gene that encodes for the large subunit of RubisCO is rbcL and detection of its expression can be used to determine the autotrophic organisms present in the environment. Recently, microarrays have been used to study functional gene expression from environmental samples such as those obtained from sediments and soil. The purpose of this thesis is to combine microarray technology and rbcL expression analysis to investigate phytoplankton populations in the Mississippi River Plume (MRP). Initially, a macroarray was constructed to determine its capabilities of quantifying gene expression in MRP. PCR amplicons were spotted onto a nylon membrane and labeled transcript RNA was hybridized to each array. Due to the large amount of cross hybridization that was observed, a microarray was used. Microarray analysis revealed large amounts of Synechococcus, pelagophyte and prymnesiophyte expression in the surface waters. Furthermore, there was no chlorophte or Prochlorococcus expression observed in the surface waters. Subsurface microarray data showed high levels of pelagophytes and other Form ID organisms. A significant chlorophyte signal was also observed in the subsurface. This study provides a third level of specificity at which phylogenetic diversity has been sampled in the MRP. Although a limited number of samples were analyzed by microarrays, this technology shows promise and this study was viewed as a pilot for their application. The rbcL probes designed were based upon published sequences from 2003 and we now have a much greater understanding of the diversity of rbcL-containing phytoplanktonic phylotypes. Future studies should employ this knowledge for judicious probe selection.

A comparison of Elysia (=Tridachia) crispata (Mörch, 1863) from the Virgin Islands with elysiid slugs from the mangrove swamps and canals in the Florida Keys that have previously been identified as E. crispata reveals many differences in habitat, gross and microscopic anatomy, food preferences of juveniles, sources of symbiotic chloroplasts and their localization within the digestive tubules, radular morphology, and nucleic acid sequences of two genes. The differences between the two groups of slugs are such that the Florida Keys animals are considered to represent a new species, Elysia clarki. Elysia clarki feeds on siphonaceous algae, and intracellularly sequesters the chloroplasts, which actively photosynthesize for up to 4 months. We have determined the algal source of the chloroplasts in adult E. clarki from 2 populations in the Florida Keys, using molecular techniques, feeding experiments, and electron microscopy. Our results clearly demonstrate that adult E. clark i sequester chloroplasts from 7 different species of algae, representing two genera, of which 5 were identified; Penicillus lamourouxii, P. capitatus, Halimeda incrassata, H. monile, and Bryopsis pennata. In addition, chloroplasts from more than 1 species of algae are sequestered in the same digestive cell simultaneously. Phylogenetic analysis of rbcL sequences from the order Bryopsidales showed that E. clarki feeding was restricted to calcareous members of the family Udoteaceae and the family Bryopsidaceae. Feeding experiments were conducted, using individuals raised in the laboratory from egg masses laid by E. clarki adults which had been collected from Grassy Key, Florida, USA, and 29 species of macroalgae. For the first 14 d post-metamorphosis, juveniles ate only the thin filamentous coenocytes, Bryopsis plumosa or Derbesia tenuissima. Electron microscopy showed that the chloroplasts from both algae were sequestered intracellularly in juvenile slugs. Individuals offered any other macroalga, including the four calcareous species fed on by adults, did not feed on or incorporate any chloroplasts, and soon died. Juveniles switched from B. plumosa to P. capitatus at a length of ~ 1.0 cm, and fixed for microscopy 14 days later had intact intracellular chloroplasts from both algae.

The oceans represent a wealth of biological diversity where many species remain to be discovered and described. Among seaweeds, a paucity of morphological features by which to differentiate species means that many genera harbour overlooked or cryptic species. Fucus, Porphyra and Ulva are three common genera of marine intertidal algae and all include species that are particularly difficult to distinguish morphologically. DNA barcoding has been championed as a revolutionary tool for species identification and discovery and applying this tool to algae was a logical step due to the difficulty of morphological identification of many algal species. This thesis is part of a significant initiative aimed at identification and discovery of all species of seaweeds in Canadian waters, using a DNA barcoding approach. The original concept of DNA barcoding relied on comparing the 5’ region of the mitochondrial cytochrome c oxidase 1 (COI-5P) gene among animal species. In this study, DNA barcoding with COI-5P was applied to the brown algal genus Fucus and worked as well as any other marker to assign morphologies to known species. The DNA barcoding results also uncovered substantial phenotypic diversity in Pacific F. distichus. Results were confirmed by comparison with sequences of the nuclear internal transcribed spacer region (ITS). For Porphyra, COI-5P DNA barcoding was compared with species identification using the chloroplast large rubisco subunit (rbcL) and the Universal Plastid Amplicon (UPA) in a floristic survey of Canadian Porphyra species. Two new species were discovered and described (Porphyra corallicola and Porphyra peggicovensis), and P. cuneiformis was synonymized with P. amplissima. The COI-5P emerged as the best marker for species discrimination despite difficulties with primer universality. To aid in choosing a marker for DNA barcoding for green algae, the universality and species discriminatory power of the rubisco large subunit (rbcL) (considering the 5’ and 3’ fragments independently), the UPA, the D2/D3 region of the nuclear large ribosomal subunit (LSU-D2/D3) and the ITS were evaluated. While the rbcL-3P highlighted several cryptic species, and worked well to distinguish Ulva species, more research is needed to recommend a marker for DNA barcoding generally in marine green macroalgae.

Variability in the genetic information DNA tracking individuals carry is easy to detect using molecular markers. In the thesis we examined related Dracaena species. For the study of genetic diversity in the genus Dracaena are used mainly noncoding regions of cpDNA, spacer trnH -- psbA, regions trnL -- trnF and trnS -- trnG -- trnG, which are more variable than coding regions. Also used in this work were coding regions of matK and rbcL. In the genus Dracaena belongs xerophytic species that are characterized by typical shaped treetop. 14 representatives of species were used in thesis growing in tropical regions of the African continent and adjacent islands and the southeastern part of the Arabian peninsula. They are monocotyledonous trees with atypical abilities of secondary thickness of trunk, which I find interesting. The massive trunk is very strong and there is potencial to used it for wood. Trees are very significant for their red plant sap which flowing from demaged trunk. The sap is very precious resource which is used in many areas of industry, for example pharmacy, traditional medicine, dye making etc. Determination of genetic affinity was based on an amplification of cpDNA template of individual Dracaena samples with primers for the studied regions. Data was obtained and evaluated by Multiple alignment program ClustalX and BioEdit after their sequencing. Evalueted data was used to create dendograms affinity. According the resulting phylogenetic tree we find out similarities and identified relationship of the monitored species of the genus Dracaena. The main purpose of research was to get answers to understand phylogenetic relationship between group of Dracaena forestry used trees. The thesis was made in cooperation of Department of Forest Botany, Dendrology and Geobiocoenology, Faculty of Forestry and Wood Technology of Mendel University in Brno.