OLIVEIRA FILHO, Rosueti Diógenes de. Incorporação de óleo essencial de manjericão em filmes biodegradáveis à base de galactomanana e óleo de canola. 2015. 77 f. Dissertação (mestrado em bioquímica)- Universidade Federal do Ceará, Fortaleza-CE, 2015 / Submitted by Elineudson Ribeiro (elineudsonr@gmail.com) on 2016-03-22T18:12:30Z No. of bitstreams: 1 2016_rdoliveirafilho.pdf: 1493866 bytes, checksum: bece9362961e66588fbfbaff693bdd5d (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-05-18T18:27:40Z (GMT) No. of bitstreams: 1 2016_rdoliveirafilho.pdf: 1493866 bytes, checksum: bece9362961e66588fbfbaff693bdd5d (MD5) / Made available in DSpace on 2016-05-18T18:27:40Z (GMT). No. of bitstreams: 1 2016_rdoliveirafilho.pdf: 1493866 bytes, checksum: bece9362961e66588fbfbaff693bdd5d (MD5) Previous issue date: 2015 / Biodegradable Packaging are used in foods as films or coatings formed by plant macromolecules such as animal, ensuring barrier properties and can serve as a carrier for antimicrobial compounds. The cell wall polysaccharides reservation, especially hemicelluloses as galactomannans, have been used widely as biodegradable polymer matrices films. The objective of this study was to evaluate the Basil essential oil (BEO) (Ocimum basilicum) incorporation in biodegradable films based on endospermic galactomannan Delonix regia seeds and canola oil (CO) (Brassica napus L.). The galactomannan (Gal) was isolated by aqueous extraction, followed by ethanol precipitation. The yield was 12.0% with high purity presented by low nitrogen content expressed protein (0.67%). The Gal showed an average intrinsic viscosity of 3.79 dL/g and molecular weight estimated in 1,04x106 g/mol. Infrared spectra by Fourier transform (FTIR) and nuclear magnetic resonance (NMR) 1H and 13C were similar to those found in the literature for other galactomannans. The fatty acid composition of CO by gas chromatography (GC-FID) showed oleic acid (62.07%), linoleic (18.91%) and heneicosanóico (6.61%) content. The major components of BEO determined by gas chromatography coupled to mass spectrometry (GC-MS) were camphor (14.85%) and 1.8-cineole (3.48%). The films were formulated with galactomannan 1% (w/v), glycerol 0.5% (w/v) and CO at different concentrations, 0.25, 0.5 and 0.75% (w/v). The films were obtained by the solvent evaporation method (casting) and characterized as the moisture content, thickness, water vapor permeability (WVP), color and opacity. The CO promoted increase of WPV and reduced moisture content when compared to the film based only in galactomannan but no significant difference between concentrations. BEO was tested against the bacterial strains of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella choleraesuis, Staphylococcus aureus and Candida albicans yeast by broth microdilution and showed minimum inhibitory concentration (MIC) ranging between 2.5 and 5.0 mg/ml and minimum bactericidal concentration (MBC) between 5.0 and 20.0 mg/ml. BEO was incorporated into film formulation with CO 0.5% (w/v) in different concentrations (2.5, 5.0 and 7.5 mg/ml). The antimicrobial films were obtained by casting and disks of 1 cm in diameter were excised and tested by the agar diffusion method and a contact zone and inhibition halos analyzed. S. aureus, E. coli and P. aeruginosa were sensitive to film containing BEO 5.0 mg/ml showed halos of 14, 15 and 16.3 mm, respectively. K. pneumoniae was inhibited only in the contact zone of the film, while S. choleraesuis and C. albicans were resistant to all formulations. However, the OC promoted changes in physical and barrier properties of the films of with galactomannan. BEO when incorporated into the film showed a moderate antimicrobial activity for potential application as bioactive packaging. / Embalagens biodegradáveis são coberturas utilizadas em alimentos na forma de filmes ou revestimentos, formadas por macromoléculas tanto de origem vegetal como de origem animal, que garantem propriedades de barreira e podem servir como veículo para compostos antimicrobianos. Os polissacarídeos de reserva de parede celular, principalmente as hemiceluloses como as galactomananas, vêm sendo extensamente usadas como matrizes poliméricas de filmes biodegradáveis. O objetivo deste trabalho foi avaliar a incorporação de óleo essencial de manjericão (OEM) (Ocimum basilicum) em filmes biodegradáveis à base de galactomanana endospérmica de sementes de flamboyant (Delonix regia) e óleo canola (OC) (Brassica napus L.). A Galactomanana (Gal) foi isolada por extração aquosa, seguida por precipitação em etanol. O rendimento determinado foi de 12,0%, com alto grau de pureza apresentado pelo baixo teor de nitrogênio expresso em proteína (0,67%). A Gal apresentou viscosidade intrínseca média de 3,79 dL/g e massa molar estimada de 1,04x106 g/mol. Os espectros de infravermelho por transformada de Fourier (FTIR) e ressonância magnética nuclear (RMN) de 1H e 13C foram similares aos encontrados na literatura para outras galactomananas. A composição em ácidos graxos do OC por cromatografia a gás (GC-FID) apresentou teores de ácido oleico (62,07%), linoleico (18,91%) e heneicosanóico (6,61%). Os componentes majoritários do OEM determinados por cromatografia a gás acoplado à espectrometria de massas (GC-MS) foram cânfora (14,85%) e 1,8-cineol (3,48%). Os filmes foram formulados com Gal 1% (m/v), glicerol 0,5% (m/v) e OC em diferentes concentrações, 0,25, 0,5 e 0,75% (m/v). Os filmes foram obtidos pelo método de evaporação de solvente (casting) e caracterizados quanto ao teor de umidade, espessura, permeabilidade ao vapor de água (PVA), cor e opacidade. O OC promoveu aumento de PVA e diminuição do teor de umidade quando comparado ao filme apenas de galactomanana, mas sem diferença significativa entre as concentrações. O OEM foi testado frente às cepas bacterianas de Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella choleraesuis, Staphylococcus aureus e levedura Candida albicans por microdiluição em caldo e apresentou concentração inibitória mínima (CIM) variando entre 2,5 e 5,0 mg/mL e concentração bactericida mínima (CBM) entre 5,0 e 20,0 mg/mL. A partir do filme com formulação de 0,5 % (m/v) de OC incorporou-se OEM em diferentes concentrações (2,5, 5,0 e 7,5 mg/mL). Os filmes antimicrobianos foram obtidos por casting e discos de 1 cm de diâmetro foram excisados e testados por método de difusão em ágar e a zona de contato e os halos de inibição analisados. S. aureus, E. coli e P. aeruginosa foram sensíveis ao filme contendo 5,0 mg/mL de OEM apresentando halos de 14, 15 e 16,3 mm, respectivamente. K. pneumoniae foi inibida apenas na zona de contato do filme, enquanto que S. choleraesuis e C. albicans foram resistentes à todas as formulações. Contudo, O OC promoveu alterações nas propriedades físicas e de barreira dos filmes de galactomanana e OEM quando incorporado ao filme apresentou uma atividade antimicrobiana moderada para potencial aplicação como embalagens bioativas.

A administração oral de isotretinoína (IST) é eficaz no tratamento da acne; no entanto, seu uso está associado a efeitos adversos, como ressecamento da pele, epistaxe, distúrbios imunológicos e psiquiátricos, enquanto a eficácia da aplicação tópica é prejudicada pela intensa irritação causada na pele. Proteger a IST do contato direto com a pele e direcionar sua liberação para os folículos pilosos, local onde a acne se desenvolve, pode reduzir a irritação e aumentar a eficácia do tratamento. O polímero Delonix (DLX), galactomanano natural derivado da semente de Delonix regia, pode ser manipulado para produzir nanopartículas para a liberação direcionada de IST na pele. Desta forma, o objetivo deste estudo foi desenvolver nanopartículas poliméricas de DLX (IST-DLX) contendo IST e avaliar seu potencial para o tratamento tópico da acne. O DLX foi coletado, purificado e caracterizado quanto a composição de monossacarídeos e massa molecular (MM). Nanopartículas DLX sem IST (branco), nanopartículas fluorescentes de DLX (nanopartículas contendo BODIPY (BOD-DLX)) e nanopartículas IST-DLX foram preparadas e caracterizadas por tamanho, índice de polidispersividade (PdI), potencial zeta (pz) e eficiência de encapsulação (EE). O estudo de liberação in vitro das nanopartículas IST-DLX foi realizado utilizando células de difusão e membrana sintética, enquanto a penetração cutânea das nanopartículas BOD-DLX e IST-DLX foi avaliada por microscopia confocal e tape stripping diferencial em pele de orelha de porco dermatomizada, respectivamente. A influência das nanopartículas IST-DLX na modulação de IL-6, IL-10 e TNF-? foi avaliada in vitro utilizando células de macrófagos AMJ-2 estimuladas com lipopolisacarídeos (LPS) e a atividade antimicrobiana foi avaliada em meio de cultura de Propionibacterium acnes. In vivo, as nanopartículas IST-DLX foram avaliadas quanto a penetração e irritação cutânea em ratos Wistar (protocolo de aprovação CEUA-FCFRP nº 17.5.213.60.3). A MM do polímero DLX foi de ~177 kDa, contendo predominantemente manose e galactose na proporção de 6,3:1. O tamanho das nanopartículas IST-DLX, determinado por dispersão de luz, foi entre 215 a 232 nm, PdI < 0,2, pz < -30 mV e EE > 25%. As nanopartículas IST-DLX sustentaram a liberação da IST, com cerca de 37% de IST sendo liberado em 48 h. As imagens de microscopia confocal mostraram que as nanopartículas BOD-DLX concentraram-se na epiderme e nos folículos pilosos em comparação com a solução de BODIPY livre, a qual permeou até a derme. Foi observada uma penetração de IST significativamente maior e descontrolada em todas as camadas da pele de orelha de porco quando a solução de polímero de IST-DLX foi usada em comparação com um acúmulo de IST menor e controlado quando as nanopartículas IST-DLX foram aplicadas. As nanopartículas IST-DLX direcionaram a IST para os folículos pilosos (26%) em comparação com a solução de IST (6,5%) que permeou por todas as camadas da pele. As nanopartículas IST-DLX reduziram significativamente a produção de IL-6 em células de macrófagos estimuladas por LPS, mesmo após 48 h, em comparação com a solução de IST livre, que reduziu significativamente a produção de IL-6 apenas até 24 h. Tanto as nanopartículas ISTDLX quanto a solução de IST livre não inibiram o crescimento de P. acnes. Não foi observado nenhum sinal de eritema nos ratos tratados com nanopartículas IST-DLX, com ou sem irradiação UVA, enquanto os tratados com solução de IST apresentaram eritema. A espessura da epiderme dos ratos tratados com as nanopartículas foi significativamente menor do que daqueles tratados com IST livre, com ou sem irradiação UVA. Conclui-se que as nanopartículas IST-DLX direcionam e sustentam a liberação de IST na pele em concentrações ótimas, modulam a resposta inflamatória da IL-6 e previnem o eritema inflamatório relacionado ao IST in vivo. Sendo assim, o polímero DLX pode ser considerado um polímero promissor para a liberação sustentada e direcionada de fármacos. / Oral isotretinoin (IST) has been effective in acne treatment; however, its use is associated with side effects such as skin dryness, epistaxis, immune disorder and psychiatric concerns while its topical gel application\'s effectiveness is hampered by irritancy. Protecting IST from direct skin contact and targeting its release to the hair follicles where the acne develops can reduce irritation and increase the effectiveness of treatment. Delonix (DLX) polymer is a natural galactomannan derived from Delonix regia seed that may be engineered to yield nanoparticles for IST skin targeting. Therefore, the aim of this study was to develop IST-loaded DLX (IST-DLX) polymeric nanoparticles and assess its skin targeting prospects for acne treatment. DLX was collected, purified and characterized by monosaccharide composition and molecular weight (Mw). Blank DLX nanoparticles, fluorescent DLX nanoparticles (BODIPY-loaded (BOD-DLX) nanoparticles) and IST-DLX nanoparticles were prepared and characterized by size, polydispersibility index (PdI), zeta potential (zp) and encapsulation efficiency (EE). In vitro release study of IST-DLX nanoparticles was performed using diffusion cells and synthetic membrane, while skin targeting of BOD-DLX and IST-DLX nanoparticles were assessed by confocal microscopy and differential tape stripping technique in dermatomed pig ear skin respectively. IST-DLX polymeric nanoparticles\' IL-6, IL-10 and TNF-? modulation was assessed in vitro using lipopolysaccharide (LPS) stimulated AMJ-2 macrophage cells and antimicrobial activity was assessed in Propionibacterium acnes culture medium. IST-DLX nanoparticles\' skin penetration and irritation were evaluated in vivo in Wistar rats (ethics committee approval protocol CEUA-FCFRP nº 17.5.213.60.3). DLX polymer Mw was ~177 kDa, containing predominantly mannose and galactose in ratio 6.3:1. IST-DLX nanoparticles\' size determined by light scattering was between 215 to 232 nm, PdI < 0.2, zp < -30 mV, EE > 25 %. IST-DLX nanoparticles sustained IST release, with about 37% IST being released in 48 h. Confocal images showed that BOD-DLX nanoparticles accumulated in the epidermis and hair follicles as compared to free BODIPY solution which permeated into the dermis. Significantly higher and uncontrolled IST penetration into all layers of pig ear skin with IST-DLX polymer solution was observed as compared to lower, controlled but optimum IST accumulation into the pig ear skin when IST-DLX nanoparticles were used. IST-DLX nanoparticles targeted the hair follicle (26 %) as compared to IST solution (6.5 %) which permeated through all skin layers. IST-DLX polymeric nanoparticles significantly reduced IL-6 production in LPS stimulated macrophage cells at 48 h as compared to free IST solution which significantly reduced IL-6 production only at 24 h. Both IST-DLX nanoparticles and free IST solution did not inhibit P. acnes growth. No sign of inflammatory erythema was observed in IST-DLX nanoparticles treated Wistar rats with or without UVA irradiation as compared to free IST treated ones. Epidermal thickness of rats treated with IST-DLX nanoparticles was significantly smaller than those treated with free IST with or without UVA irradiation. It can be concluded that IST-DLX nanoparticles can target, deliver and sustain IST release to the skin at optimum concentrations, modulate IL-6 inflammatory responses and may prevent IST related inflammatory erythema in vivo. DLX polymer could be promising polymer for drug delivery.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Embalagens biodegradÃveis sÃo coberturas utilizadas em alimentos na forma de filmes ou revestimentos, formadas por macromolÃculas tanto de origem vegetal como de origem animal, que garantem propriedades de barreira e podem servir como veÃculo para compostos antimicrobianos. Os polissacarÃdeos de reserva de parede celular, principalmente as hemiceluloses como as galactomananas, vÃm sendo extensamente usadas como matrizes polimÃricas de filmes biodegradÃveis. O objetivo deste trabalho foi avaliar a incorporaÃÃo de Ãleo essencial de manjericÃo (OEM) (Ocimum basilicum) em filmes biodegradÃveis à base de galactomanana endospÃrmica de sementes de flamboyant (Delonix regia) e Ãleo canola (OC) (Brassica napus L.). A Galactomanana (Gal) foi isolada por extraÃÃo aquosa, seguida por precipitaÃÃo em etanol. O rendimento determinado foi de 12,0%, com alto grau de pureza apresentado pelo baixo teor de nitrogÃnio expresso em proteÃna (0,67%). A Gal apresentou viscosidade intrÃnseca mÃdia de 3,79 dL/g e massa molar estimada de 1,04x106 g/mol. Os espectros de infravermelho por transformada de Fourier (FTIR) e ressonÃncia magnÃtica nuclear (RMN) de 1H e 13C foram similares aos encontrados na literatura para outras galactomananas. A composiÃÃo em Ãcidos graxos do OC por cromatografia a gÃs (GC-FID) apresentou teores de Ãcido oleico (62,07%), linoleico (18,91%) e heneicosanÃico (6,61%). Os componentes majoritÃrios do OEM determinados por cromatografia a gÃs acoplado à espectrometria de massas (GC-MS) foram cÃnfora (14,85%) e 1,8-cineol (3,48%). Os filmes foram formulados com Gal 1% (m/v), glicerol 0,5% (m/v) e OC em diferentes concentraÃÃes, 0,25, 0,5 e 0,75% (m/v). Os filmes foram obtidos pelo mÃtodo de evaporaÃÃo de solvente (casting) e caracterizados quanto ao teor de umidade, espessura, permeabilidade ao vapor de Ãgua (PVA), cor e opacidade. O OC promoveu aumento de PVA e diminuiÃÃo do teor de umidade quando comparado ao filme apenas de galactomanana, mas sem diferenÃa significativa entre as concentraÃÃes. O OEM foi testado frente Ãs cepas bacterianas de Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella choleraesuis, Staphylococcus aureus e levedura Candida albicans por microdiluiÃÃo em caldo e apresentou concentraÃÃo inibitÃria mÃnima (CIM) variando entre 2,5 e 5,0 mg/mL e concentraÃÃo bactericida mÃnima (CBM) entre 5,0 e 20,0 mg/mL. A partir do filme com formulaÃÃo de 0,5 % (m/v) de OC incorporou-se OEM em diferentes concentraÃÃes (2,5, 5,0 e 7,5 mg/mL). Os filmes antimicrobianos foram obtidos por casting e discos de 1 cm de diÃmetro foram excisados e testados por mÃtodo de difusÃo em Ãgar e a zona de contato e os halos de inibiÃÃo analisados. S. aureus, E. coli e P. aeruginosa foram sensÃveis ao filme contendo 5,0 mg/mL de OEM apresentando halos de 14, 15 e 16,3 mm, respectivamente. K. pneumoniae foi inibida apenas na zona de contato do filme, enquanto que S. choleraesuis e C. albicans foram resistentes à todas as formulaÃÃes. Contudo, O OC promoveu alteraÃÃes nas propriedades fÃsicas e de barreira dos filmes de galactomanana e OEM quando incorporado ao filme apresentou uma atividade antimicrobiana moderada para potencial aplicaÃÃo como embalagens bioativas. / Biodegradable Packaging are used in foods as films or coatings formed by plant macromolecules such as animal, ensuring barrier properties and can serve as a carrier for antimicrobial compounds. The cell wall polysaccharides reservation, especially hemicelluloses as galactomannans, have been used widely as biodegradable polymer matrices films. The objective of this study was to evaluate the Basil essential oil (BEO) (Ocimum basilicum) incorporation in biodegradable films based on endospermic galactomannan Delonix regia seeds and canola oil (CO) (Brassica napus L.). The galactomannan (Gal) was isolated by aqueous extraction, followed by ethanol precipitation. The yield was 12.0% with high purity presented by low nitrogen content expressed protein (0.67%). The Gal showed an average intrinsic viscosity of 3.79 dL/g and molecular weight estimated in 1,04x106 g/mol. Infrared spectra by Fourier transform (FTIR) and nuclear magnetic resonance (NMR) 1H and 13C were similar to those found in the literature for other galactomannans. The fatty acid composition of CO by gas chromatography (GC-FID) showed oleic acid (62.07%), linoleic (18.91%) and heneicosanÃico (6.61%) content. The major components of BEO determined by gas chromatography coupled to mass spectrometry (GC-MS) were camphor (14.85%) and 1.8-cineole (3.48%). The films were formulated with galactomannan 1% (w/v), glycerol 0.5% (w/v) and CO at different concentrations, 0.25, 0.5 and 0.75% (w/v). The films were obtained by the solvent evaporation method (casting) and characterized as the moisture content, thickness, water vapor permeability (WVP), color and opacity. The CO promoted increase of WPV and reduced moisture content when compared to the film based only in galactomannan but no significant difference between concentrations. BEO was tested against the bacterial strains of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella choleraesuis, Staphylococcus aureus and Candida albicans yeast by broth microdilution and showed minimum inhibitory concentration (MIC) ranging between 2.5 and 5.0 mg/ml and minimum bactericidal concentration (MBC) between 5.0 and 20.0 mg/ml. BEO was incorporated into film formulation with CO 0.5% (w/v) in different concentrations (2.5, 5.0 and 7.5 mg/ml). The antimicrobial films were obtained by casting and disks of 1 cm in diameter were excised and tested by the agar diffusion method and a contact zone and inhibition halos analyzed. S. aureus, E. coli and P. aeruginosa were sensitive to film containing BEO 5.0 mg/ml showed halos of 14, 15 and 16.3 mm, respectively. K. pneumoniae was inhibited only in the contact zone of the film, while S. choleraesuis and C. albicans were resistant to all formulations. However, the OC promoted changes in physical and barrier properties of the films of with galactomannan. BEO when incorporated into the film showed a moderate antimicrobial activity for potential application as bioactive packaging.

Made available in DSpace on 2014-08-20T13:44:28Z (GMT). No. of bitstreams: 1 dissertacao_claudinei_antonio_bolognez.pdf: 182881 bytes, checksum: 028c23054f130496cda513a0975aee69 (MD5) Previous issue date: 2012-05-25 / The Flamboyant (Delonix regia (Bojer ex Hook) Raf.) is an exotic ornamental species adapted to the conditions of the Brazilian territory, belonging to subfamily Caesalpinioideae and it has seeds with hard coat that promotes seed dormancy caused by the impermeability of seed coat to water and oxygen. The aim of this study was to test different types of mechanical scarification to overcome seedcoat dormancy of seed, associated with physical and biological aspects of Flamboyant. The study was conducted in laboratory and experimental area of the Department of Biological Sciences at the Institute of Exact and Natural Sciences, Federal University of Mato Grosso, in Rondonópolis, MT, from October 2011 to April 2012, the experimental field was protected by a screen shading retaining 50% of brightness. The treatments were: control, whose seeds had not been scarified, treatments with a hole in the coat, mechanical scarification with sandpaper number 150 for wood, water heated to 90 ° C with immersion of the seeds and water heating to 90 ° C and then immersion seeds leaving to cool to room temperature. Experimental design was completely randomized (CRD) with five treatments and four repetitions, totaling twenty installments of fifteen seeds each. With the observations, there were no significant differences between the means of overcoming the five methods employed in the work. To overcome dormancy in flamboyant the recommended method is immersion in water heated to 90 ° C and subsequent soaking of the seeds. The use of techniques to overcome flamboyant seeds may cause problems when growth in height and diameter of plants in the initial phase of establishment. / O Flamboyant (Delonix regia (Bojer ex Hook) Raf.) é uma espécie ornamental exótica adaptada as condições do território brasileiro, pertencente á subfamília Caesalpinioideae e apresenta sementes com tegumento duro que favorece a dormência das sementes causada pela impermeabilidade do tegumento à água e ao oxigênio. O objetivo deste estudo foi testar diferentes modalidades de escarificação mecânica para superar a dormência tegumentar da semente, associada a aspectos físicos e biológicos de Flamboyant. O trabalho foi desenvolvido em laboratório e área experimental pertencente ao Departamento de Ciências Biológicas do Instituto de Ciências Exatas e Naturais, Universidade Federal de Mato Grosso, em Rondonópolis, MT, no período de outubro de 2011 a abril de 2012, a área experimental de campo era protegida por uma tela sombrite retendo 50% da luminosidade. Os tratamentos foram os seguintes: testemunha cujas sementes não sofreram escarificação, tratamentos com furo no tegumento, escarificação mecânica com lixa nº 150 para madeira, água aquecida a 90°C com a imersão das sementes e aquecimento da água até 90°C e posterior imersão das sementes deixando esfriar até temperatura ambiente. Delineamento utilizado foi Inteiramente Casualizado (DIC), com cinco tratamentos e quatro repetições, totalizando vinte parcelas de quinze sementes cada. Com as observações, não foram detectadas diferença significativa entre as médias nos cinco métodos de superação empregados no trabalho. Para superação da dormência em flamboyant o método recomendado é a imersão em água aquecida até 90°C e posterior imersão das sementes. O uso das técnicas de superação de sementes de flamboyant pode causar problemas ao crescimento em altura e diâmetro das plantas, na fase inicial de estabelecimento.

Despite their ecological and economic importance, the majority of plant species and their conservation status are poorly known. Only 4% of plants have been assessed globally and listed on the IUCN Red List of Threatened Species; and without plant conservation assessments, many plant species will not feature in conservation planning. Herbarium collection information can significantly increase the number of plant conservation assessments. Thus, the aims of this thesis were: (1) to investigate how the quality of herbarium-based conservation assessments can be optimised; (2) to assess the extent to which herbarium-based conservation assessments reflect the reality on the ground; and (3) to scientifically validate genetic and spatial underpinning of IUCN criteria. Preliminary range-based assessments of the Leguminosae of Madagascar achieved a result consistent with the final conservation rating for over 95% of species when using up to fifteen herbarium specimens. Bioclimatic modelling of range shifts based on future climate change predicted that, in the worst case scenario, up to one third of endemic Leguminosae in Madagascar will be threatened with extinction over the next 100 years. An analysis of the population structure of species of Delonix s.l. (Leguminosae) showed that combining spatial analysis with population genetic data provides a more complete picture of landscape-level population dynamics and the impacts on conservation status. Moreover, range-wide genetic analysis of AFLP markers for four species of Delonix demonstrated a genetic basis for IUCN categories distinguishing between threatened and non-threatened species. Although genetic data are currently not often incorporated in conservation assessments, they are crucial in making accurate management decisions and creating effective action plans for conservation. Only by using all available scientific resources can informed conservation decisions be made and the survival of plants and their associated ecosystems be ensured.

Ce mémoire porte sur les relations phylogénétiques, géographiques et historiques du genre afro-malgache Delonix qui contient onze espèces et des genres monospécifiques et endémiques Colvillea et Lemuropisum. Les relations intergénériques et interspécifiques entre les espèces de ces trois genres ne sont pas résolues ce qui limite la vérification d’hypothèses taxonomiques, mais également biogéographiques concernant la dispersion de plantes depuis ou vers Madagascar. Une meilleure compréhension des relations évolutives et biogéographiques entre ces espèces menacées d’extinction permettrait une plus grande efficacité quant à leur conservation. L’objectif de ce mémoire est de reconstruire la phylogénie des espèces à l’aide de régions moléculaires des génomes chloroplastique et nucléaire, d’identifier les temps de divergences entre les espèces et de reconstruire l’aire géographique ancestrale pour chacun des groupes. Ce projet démontre que le genre Delonix n’est pas soutenu comme étant monophylétique et qu’une révision taxonomique s’impose. Les relations intergénériques demeurent floues quant à la position phylogénétique de Colvillea et nos résultats suggèrent de l’hybridation ou un assortiment incomplet de cette lignée. Les espèces sont apparues et se sont diversifiées au Miocène à partir d’un ancêtre commun du sud de Madagascar. La phylogénie montre deux clades associés aux aires géographiques de répartition des espèces opposant les espèces largement répandues à celles majoritairement restreintes au fourré aride. Différentes hypothèses afin d’expliquer la dispersion des Delonix africains au Miocène à partir de Madagascar sont discutées. Un point de mire sur les interactions biotiques et abiotiques, passées et présentes, dans le fourré aride de Madagascar est recommandé en terme de conservation. / This thesis discusses the phylogenetic, geographic, and historic relationships of the eleven species of the afro-madagascan genus Delonix, as well as the endemic, monospecific genera Colvillea and Lemuropisum. The intergeneric and interspecific relationships of these taxa are unresolved which limits the evaluation of taxonomic changes suggested by some studies and of conflicting biogeographical hypotheses regarding plant dispersal around Madagascar. These species are all endangered, therefore a better understanding of their evolutionary pattern would benefit conservation efforts in Madagascar. The objective of this thesis is to reconstruct a phylogeny using multiple loci from two distinct genomes, and to use it to reconstruct the ancestral geographic range of species and estimate the times of divergence of the lineages. Results indicate that genus Delonix is not supported as monophyletic and a taxonomic revision is necessary. The exact position for Colvillea is ambiguous and suggests hybridization or incomplete lineage sorting for this taxon. Species evolved mostly during the Miocene from southern Madagascan ancestors. The phylogeny shows two clades associated with the geographic distribution of species, opposing the widespread species to the rare and mostly southern, endemic, spiny thicket species. The different hypotheses that could explain the Miocene long distance dispersal of the two African Delonix species from Madagascar are discussed. We recommend that conservation efforts should focus on past and present biotic and abiotic interactions in the Madagascar spiny thicket, considering that this unique habitat is an evolutionary cradle for at least three genera.

Ce mémoire porte sur les relations phylogénétiques, géographiques et historiques du genre afro-malgache Delonix qui contient onze espèces et des genres monospécifiques et endémiques Colvillea et Lemuropisum. Les relations intergénériques et interspécifiques entre les espèces de ces trois genres ne sont pas résolues ce qui limite la vérification d’hypothèses taxonomiques, mais également biogéographiques concernant la dispersion de plantes depuis ou vers Madagascar. Une meilleure compréhension des relations évolutives et biogéographiques entre ces espèces menacées d’extinction permettrait une plus grande efficacité quant à leur conservation. L’objectif de ce mémoire est de reconstruire la phylogénie des espèces à l’aide de régions moléculaires des génomes chloroplastique et nucléaire, d’identifier les temps de divergences entre les espèces et de reconstruire l’aire géographique ancestrale pour chacun des groupes. Ce projet démontre que le genre Delonix n’est pas soutenu comme étant monophylétique et qu’une révision taxonomique s’impose. Les relations intergénériques demeurent floues quant à la position phylogénétique de Colvillea et nos résultats suggèrent de l’hybridation ou un assortiment incomplet de cette lignée. Les espèces sont apparues et se sont diversifiées au Miocène à partir d’un ancêtre commun du sud de Madagascar. La phylogénie montre deux clades associés aux aires géographiques de répartition des espèces opposant les espèces largement répandues à celles majoritairement restreintes au fourré aride. Différentes hypothèses afin d’expliquer la dispersion des Delonix africains au Miocène à partir de Madagascar sont discutées. Un point de mire sur les interactions biotiques et abiotiques, passées et présentes, dans le fourré aride de Madagascar est recommandé en terme de conservation. / This thesis discusses the phylogenetic, geographic, and historic relationships of the eleven species of the afro-madagascan genus Delonix, as well as the endemic, monospecific genera Colvillea and Lemuropisum. The intergeneric and interspecific relationships of these taxa are unresolved which limits the evaluation of taxonomic changes suggested by some studies and of conflicting biogeographical hypotheses regarding plant dispersal around Madagascar. These species are all endangered, therefore a better understanding of their evolutionary pattern would benefit conservation efforts in Madagascar. The objective of this thesis is to reconstruct a phylogeny using multiple loci from two distinct genomes, and to use it to reconstruct the ancestral geographic range of species and estimate the times of divergence of the lineages. Results indicate that genus Delonix is not supported as monophyletic and a taxonomic revision is necessary. The exact position for Colvillea is ambiguous and suggests hybridization or incomplete lineage sorting for this taxon. Species evolved mostly during the Miocene from southern Madagascan ancestors. The phylogeny shows two clades associated with the geographic distribution of species, opposing the widespread species to the rare and mostly southern, endemic, spiny thicket species. The different hypotheses that could explain the Miocene long distance dispersal of the two African Delonix species from Madagascar are discussed. We recommend that conservation efforts should focus on past and present biotic and abiotic interactions in the Madagascar spiny thicket, considering that this unique habitat is an evolutionary cradle for at least three genera.