A 1,3-dimetilamilamina (DMAA) é um estimulante que passou a ser adicionada aos suplementos alimentares e compostos emagrecedores a partir de 2006, sendo amplamente consumida por atletas e militares americanos. No entanto, após relatos de toxicidade a DMAA foi proibida por agências regulatórias do Brasil e Estados Unidos. Porém, mesmo após a sua proibição, a DMAA ainda pode ser encontrada em suplementos alimentares. A sua origem foi relacionada ao óleo essencial de Pelargonium graveolens, e, no entanto, inúmeros autores questionaram os resultados originais e a sua origem natural. Adicionalmente, os óleos essenciais de espécies de Pelargonium tiveram a sua atividade antimicrobiana reportada frente a bactérias e fungos. Assim, considerando os aspectos abordados, este trabalho teve como objetivo determinar a presença de DMAA nos óleos essenciais de Pelargonium spp. por GC-MS, DART-MS/MS e LC-MS/MS; assim como nas folhas das mesmas espécies, utilizando a extração por headspace, previamente otimizada, seguida de análise por GC-MS. Também se propôs a investigar a atividade antifúngica dos óleos essenciais de P. graveolens de diferentes origens e desenvolver uma formulação contendo uma nanoemulsão do óleo para o tratamento de candidíase vaginal. Por fim, teve como objetivo desenvolver metodologia de screening para avaliar a presença de DMAA e outros estimulantes em suplementos alimentares apreendidos, através de DART-MS/MS. Os resultados revelaram que a DMAA não está presente nos óleos essenciais de diferentes espécies de Pelargonium spp. obtidos por hidrodestilação, do Rio Grande do Sul. Após a otimização através de desenho experimental, a técnica de headspace provou ser eficaz na extração dos constituintes voláteis presentes nas folhas e, no entanto, a DMAA não foi detectada, assim como nos óleos essenciais comerciais de P. graveolens do Brasil, China, Egito, África do Sul, Albânia e Ilhas Reunião. Os óleos essenciais apresentaram atividade antifúngica frente às cinco espécies de Candida. Ainda, este efeito antifúngico apresentou melhores resultados com a nanoformulação contendo o óleo essencial. A análise de screening por DART-MS/MS se mostrou eficaz na detecção de DMAA, efedrina, sinefrina, cafeína, sibutramina e metilfenidato, em amostras de suplementos alimentares apreendidos, apresentando resultados positivos para todos os estimulantes. Com base nos resultados obtidos e nos objetivos propostos, verificou-se que mesmo após a 12 utilização de três técnicas analíticas distintas e uma nova alternativa para extração dos constituintes voláteis, a DMAA não foi econtrada nos óleos essenciais e nas folhas das espécies de Pelargonium, corroborando com outros estudos realizados, e indicando que a sua origem não é natural nestas espécies. A formulação final contendo a nanoemulsão com o óleo essencial apresentou atividade antifúngica superior a do óleo essencial livre. As análises das amostras apreendidas mostraram que mesmo após a sua proibição pelas agências regulatórias, os suplementos contendo DMAA e outros estimulantes ainda são comercializados, representando um grande risco para a saúde dos seus usuários. / 1,3-dimethylamylamine (DMAA) is a stimulant that started to be added in dietary supplements and weight loss compounds since 2006 and is widely consumed by athletes and the USA army. However, after reports of toxicity DMAA has been banned by regulatory agencies in Brazil and United States. However, even after its prohibition, DMAA still can be found in dietary supplements. Its origin was related to the essential oils of Pelargonium graveolens, and, however, many authors questioned the results and its natural origin. In addition, the essential oils of species of Pelargonium, had their antimicrobial activity reported against bacteria and fungi. Considering the aspects mentioned, this work aimed to determine the presence of DMAA in the essential oils by GC-MS, DART-MS/MS and LC-MS/MS; as well as in the leaves of the same species using the headspace extraction, previously optimized, followed by analysis through GC-MS. It has also been proposed to investigate the antifungal activity of essential oils of P. graveolens from different origins and develop a formulation containing an oil nanoemulsion for the treatment of vaginal candidiasis. Finally, it aimed to develop a screening method to evaluate the presence of DMAA and other stimulants in seized dietary supplements by DART-MS/MS. The results showed that DMAA is not present in the Rio Grande do Sul’s essential oils of Pelargonium spp. obtained by hydrodistillation. After optimization through experimental design, the headspace technique proved to be effective in extracting volatile constituents present in the leaves and, however, DMAA was not detected, as well as in commercial essential oils of P. graveolens from Brazil, China, Egypt, South Africa, Albania and Reunion Islands. The essential oils presented antifungal activity against five Candida species. Furthermore, this antifungal effect presented better results with the nanoformulation containing essential oil. DART-MS/MS screening was effective in detection of DMAA, ephedrine, synephrine, caffeine, sibutramine and methylphenidate in seized dietary supplements, showing positive results for all stimulants. Based on the results obtained and proposed objectives, it was verified that even after using three different analytical techniques and a new alternative for volatile constituents extraction, DMAA was not found in essential oils and leaves of Pelargonium spp., corroborating with other studies carried out, and indicating that its origin is not natural in these species. The final formulation containing the nanoemulsion with the essential oil had antifungal activity superior compared to 14 dispersed essential oil. The analysis of seized samples showed that even after its prohibition by regulatory agencies, supplements containing DMAA and other stimulants are still commercialized, representing a major health risk for their users.

Plastid genomes of flowering plants are largely identical in gene order and content, but a few lineages have been identified with many gene and intron losses, genomic rearrangements, and accelerated rates of nucleotide substitutions. These aberrant lineages present an opportunity to understand the modes of selection acting on these genomes as well as their long-term stability. My research has focused on two areas within plastid genome evolution in Geraniaceae: first, an investigation of the diversity of unusual plastid genomes in a single genus, Erodium (Geraniaceae) for chapters one and three. Chapter two focuses on the evolution of subunits of the plastid-encoded RNA polymerase (PEP). The first chapter described the loss of plastid-encoded NADPH dehydrogenase (ndh) genes from a clade of 13 Erodium species. Divergence time estimates indicate this clade is less than 5 million years old. This recent loss of ndh genes in Erodium presents an opportunity to investigate changes in photosynthetic function through comparative biochemistry between Erodium species with and without plastid-encoded ndh genes. Second, I examined the evolution of the gene encoding the alpha subunit (rpoA) of PEP in three disparate angiosperm lineages—Pelargonium (Geraniaceae), Passiflora (Passifloraceae), and Annonaceae—in which this gene has diverged so greatly that it is barely recognizable. PEP is conserved in the plastid genomes of all photosynthetic angiosperms. I found multiple lines of evidence indicating that the genes remain functional despite retaining only ~30% sequence identity with rpoA genes from outgroups. The genomes containing these divergent rpoA genes have undergone significant rearrangement due to illegitimate recombination and gene conversion, and I hypothesized that these phenomena have also driven the divergence of rpoA. Third, I conducted a survey of plastid genome evolution in Erodium with the completion of 15 additional whole genomes. Except for Erodium and some legumes, all angiosperm plastid genomes share a quadripartite structure with large and small single copy regions (LSC, SSC) and two inverted repeats (IR). I discovered a species of Erodium that has re-formed a large inverted repeat. Demonstrating a precedent for loss and regain of the IR also impacts models of evolution for other highly rearranged plastid genomes. / text

A 1,3-dimetilamilamina (DMAA) é um estimulante que passou a ser adicionada aos suplementos alimentares e compostos emagrecedores a partir de 2006, sendo amplamente consumida por atletas e militares americanos. No entanto, após relatos de toxicidade a DMAA foi proibida por agências regulatórias do Brasil e Estados Unidos. Porém, mesmo após a sua proibição, a DMAA ainda pode ser encontrada em suplementos alimentares. A sua origem foi relacionada ao óleo essencial de Pelargonium graveolens, e, no entanto, inúmeros autores questionaram os resultados originais e a sua origem natural. Adicionalmente, os óleos essenciais de espécies de Pelargonium tiveram a sua atividade antimicrobiana reportada frente a bactérias e fungos. Assim, considerando os aspectos abordados, este trabalho teve como objetivo determinar a presença de DMAA nos óleos essenciais de Pelargonium spp. por GC-MS, DART-MS/MS e LC-MS/MS; assim como nas folhas das mesmas espécies, utilizando a extração por headspace, previamente otimizada, seguida de análise por GC-MS. Também se propôs a investigar a atividade antifúngica dos óleos essenciais de P. graveolens de diferentes origens e desenvolver uma formulação contendo uma nanoemulsão do óleo para o tratamento de candidíase vaginal. Por fim, teve como objetivo desenvolver metodologia de screening para avaliar a presença de DMAA e outros estimulantes em suplementos alimentares apreendidos, através de DART-MS/MS. Os resultados revelaram que a DMAA não está presente nos óleos essenciais de diferentes espécies de Pelargonium spp. obtidos por hidrodestilação, do Rio Grande do Sul. Após a otimização através de desenho experimental, a técnica de headspace provou ser eficaz na extração dos constituintes voláteis presentes nas folhas e, no entanto, a DMAA não foi detectada, assim como nos óleos essenciais comerciais de P. graveolens do Brasil, China, Egito, África do Sul, Albânia e Ilhas Reunião. Os óleos essenciais apresentaram atividade antifúngica frente às cinco espécies de Candida. Ainda, este efeito antifúngico apresentou melhores resultados com a nanoformulação contendo o óleo essencial. A análise de screening por DART-MS/MS se mostrou eficaz na detecção de DMAA, efedrina, sinefrina, cafeína, sibutramina e metilfenidato, em amostras de suplementos alimentares apreendidos, apresentando resultados positivos para todos os estimulantes. Com base nos resultados obtidos e nos objetivos propostos, verificou-se que mesmo após a 12 utilização de três técnicas analíticas distintas e uma nova alternativa para extração dos constituintes voláteis, a DMAA não foi econtrada nos óleos essenciais e nas folhas das espécies de Pelargonium, corroborando com outros estudos realizados, e indicando que a sua origem não é natural nestas espécies. A formulação final contendo a nanoemulsão com o óleo essencial apresentou atividade antifúngica superior a do óleo essencial livre. As análises das amostras apreendidas mostraram que mesmo após a sua proibição pelas agências regulatórias, os suplementos contendo DMAA e outros estimulantes ainda são comercializados, representando um grande risco para a saúde dos seus usuários. / 1,3-dimethylamylamine (DMAA) is a stimulant that started to be added in dietary supplements and weight loss compounds since 2006 and is widely consumed by athletes and the USA army. However, after reports of toxicity DMAA has been banned by regulatory agencies in Brazil and United States. However, even after its prohibition, DMAA still can be found in dietary supplements. Its origin was related to the essential oils of Pelargonium graveolens, and, however, many authors questioned the results and its natural origin. In addition, the essential oils of species of Pelargonium, had their antimicrobial activity reported against bacteria and fungi. Considering the aspects mentioned, this work aimed to determine the presence of DMAA in the essential oils by GC-MS, DART-MS/MS and LC-MS/MS; as well as in the leaves of the same species using the headspace extraction, previously optimized, followed by analysis through GC-MS. It has also been proposed to investigate the antifungal activity of essential oils of P. graveolens from different origins and develop a formulation containing an oil nanoemulsion for the treatment of vaginal candidiasis. Finally, it aimed to develop a screening method to evaluate the presence of DMAA and other stimulants in seized dietary supplements by DART-MS/MS. The results showed that DMAA is not present in the Rio Grande do Sul’s essential oils of Pelargonium spp. obtained by hydrodistillation. After optimization through experimental design, the headspace technique proved to be effective in extracting volatile constituents present in the leaves and, however, DMAA was not detected, as well as in commercial essential oils of P. graveolens from Brazil, China, Egypt, South Africa, Albania and Reunion Islands. The essential oils presented antifungal activity against five Candida species. Furthermore, this antifungal effect presented better results with the nanoformulation containing essential oil. DART-MS/MS screening was effective in detection of DMAA, ephedrine, synephrine, caffeine, sibutramine and methylphenidate in seized dietary supplements, showing positive results for all stimulants. Based on the results obtained and proposed objectives, it was verified that even after using three different analytical techniques and a new alternative for volatile constituents extraction, DMAA was not found in essential oils and leaves of Pelargonium spp., corroborating with other studies carried out, and indicating that its origin is not natural in these species. The final formulation containing the nanoemulsion with the essential oil had antifungal activity superior compared to 14 dispersed essential oil. The analysis of seized samples showed that even after its prohibition by regulatory agencies, supplements containing DMAA and other stimulants are still commercialized, representing a major health risk for their users.

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2017-06-01T16:11:21Z No. of bitstreams: 1 texto completo.pdf: 487232 bytes, checksum: 4fd53655f2412899670b34bd2a57487e (MD5) / Made available in DSpace on 2017-06-01T16:11:21Z (GMT). No. of bitstreams: 1 texto completo.pdf: 487232 bytes, checksum: 4fd53655f2412899670b34bd2a57487e (MD5) Previous issue date: 2004-08-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Este trabalho teve como objetivos: - estudar a sensibilidade ao etileno de duas variedades de gerânio em vaso durante sua fase pós- produção; - investigar a eficiência do 1-metilciclopropeno (1-MCP) na prevenção dos danos causados pelo etileno e determinar seus melhores níveis para a aplicação em vasos de gerânio visando o aumento da vida de prateleira e diminuição da queda de pétalas. O experimento sensibilidade ao etileno exógeno foi instalado em parcelas subdivididas, tendo nas parcelas 5 avaliações, uma antes (0 h) e 4 após o início da exposição ao etileno (3, 24, 48 e 72 h) e nas subparcelas um esquema fatorial 3x2, com 3 concentrações de etileno (0, 10 e 100 μL/L) e 2 variedades, no DIC com 3 repetições e 1 vaso de gerânio por repetição. As plantas, das variedades Pulsar Red (VR) e Pulsar Salmon (VS), foram expostas ao etileno em câmaras herméticas, por 3 h. A queda de pétalas (QP) foi avaliada logo após o tratamento e a cada 24 h até todas as plantas atingirem 100% de QP. No experimento com 1- MCP utilizou-se um esquema de parcelas subdivididas, com 4 tempos de exposição ao 1-MCP nas parcelas (3, 6, 9 e 12 h) e, nas subparcelas, um esquema fatorial 5x2, com 5 concentrações de Ethylbloc® (0; 0,1; 0,5; 1,0 e 1,5 g/m3) e 2 variedades, com 3 repetições e 1 vaso de gerânio por repetição. Após o tratamento com 1-MCP as plantas foram expostas a 1 μL/L de etileno, durante 3 h. Avaliou-se o número de dias para queda de 50% das pétalas (D50) e a vida de prateleira da planta. No primeiro experimento, 3 horas após o início da exposição a 10 μL/L de etileno, observou-se diferença (P < 0,05) entre as duas variedades (VR = 97,3% e VS = 76,0%). Após 24 h, plantas da VR e VS sem exposição ao etileno, apresentaram diferença (P < 0,05) (VR = 76% e VS = 42,7%). A % QP foi afetada significativamente pelo aumento da concentração de etileno, sendo mais precoce em plantas tratadas com o gás. Já no segundo experimento a vida de prateleira teve média maior (P < 0,05) na VR (30,3 dias) que VS (12,7 dias) em 12 h e 1 g/m3 de Ethylbloc®. O número de dias para 50% de queda de pétalas atingiu as maiores médias na VR, sendo diferente da VS, com 3 h e 1 μL/L (7,7 e 4,3 dias, respectivamente). Analisando-se os dados nas equações de regressão, foram obtidos os valores máximos de vida de prateleira para VR (23,98 dias) com 3 h e 0,84 g/m3 de Ethylbloc®. A equação de regressão de D50 para VR tem valores máximos com 3,56 h e 1,5 g/m3 de Ethylbloc® (6,9 dias). D50 da VS não foi influenciado pelos níveis de concentração de 1-MCP e pelo tempo de exposição, apresentando média de 4,6 dias. Com o experimento exposição a etileno exógeno, conclui- se que as duas variedades respondem à presença de 10 μL/L de etileno exógeno, por 3 h, mas a VR é mais sensível que a VS. Com o experimento exposição ao 1-MCP, conclui-se que a melhor combinação entre concentração e tempo de exposição para inibir a queda precoce de pétalas e aumentar a vida de prateleira da VR é 1,0 g/m3 e 3 h, e que para a VS, o 1- MCP não foi eficiente. / The objectives of this dissertation were to study the ethylene sensitivity of two varieties of potted geranium during post-production, to evaluate the efficacy of 1-MCP in preventing ethylene-damages, and the optimum exposure time and concentration of 1-MCP to increase shelf life and overcome petal abscission. The first experiment was installed in split-plot pattern with 5 evaluations after the beginning of ethylene treatment (0, 3, 24, 48 and 72 h) in the main plot and the sub plot with a factorial pattern 3x2 in completely randomized factorial - 3 ethylene concentrations (0, 10 and 100 μL/L) and 2 varieties (Pulsar Red, VR, and Pulsar Salmon, VS), with 3 replicates each. The plants were exposed to ethylene in closed chambers, for 3 h. Petal abscission (PA) was evaluated just after the treatment with 24 h intervals until reached 100% PA. The second experiment had the experimental design in split-plots, with 4 exposure times (3, 6, 9 e 12 h) in the main plot and a factorial pattern 5x2 in completely randomized factorial in the sub plot - 5 Ethylbloc® concentrations (0, 0.1; 0.5; 1.0 e 1.5 g/m3) and 2 varieties (VR e VS), with 3 replicates each. After the 1-MCP treatment, the plants were exposed to 1 μL/L of ethylene, for 3 h. The number of days to 50% petal abscission (D50) and the shelf life (C30) were evaluated. Three hours after beginning the first experiment, 10 μL/L of ethylene caused different (P < 0.05) % PA between the two varieties (VR = 97.3% and VS = 76.0%). After 24 h, plants of VR and VS without exposition to ethylene presented difference (VR = 76% and VS = 42.7%) in petal abscission. The % PA was affected significantly by the increase of the ethylene concentration, being higher in plants exposed to ethylene. In the second experiment, shelf life was higher (P < 0.05) to VR (30.3 days) than VS (12.7 days) with 12 h and 1 g/m3 of Ethylbloc®. D50 reached higher value for VR, being different from VS, with 7.7 e 4.3 days, respectively. Based on data regression analysis, maximum values for C30 in VR (23.98 days) were found with 3 h and 0.84 g/m3 of Ethylbloc. The D50 regression for VR has maximum values with 3.56 h and 1.5 g/m3 of Ethylbloc (6.91 days). D50 of VS was not influenced by the treatment with 1-MCP, and presented average of 4.55 days. It can be concluded that VR is more sensitive to ethylene than VS, but the two varieties respond to the presence of 10 μL/L of ethylene, for 3 h; the best combination between concentration and treatment length to inhibit petal abscission and to increase the shelf life of VR was 1.0 g/m3 for 3 h and the treatment with 1-MCP in VS was not efficient.

A 1,3-dimetilamilamina (DMAA) é um estimulante que passou a ser adicionada aos suplementos alimentares e compostos emagrecedores a partir de 2006, sendo amplamente consumida por atletas e militares americanos. No entanto, após relatos de toxicidade a DMAA foi proibida por agências regulatórias do Brasil e Estados Unidos. Porém, mesmo após a sua proibição, a DMAA ainda pode ser encontrada em suplementos alimentares. A sua origem foi relacionada ao óleo essencial de Pelargonium graveolens, e, no entanto, inúmeros autores questionaram os resultados originais e a sua origem natural. Adicionalmente, os óleos essenciais de espécies de Pelargonium tiveram a sua atividade antimicrobiana reportada frente a bactérias e fungos. Assim, considerando os aspectos abordados, este trabalho teve como objetivo determinar a presença de DMAA nos óleos essenciais de Pelargonium spp. por GC-MS, DART-MS/MS e LC-MS/MS; assim como nas folhas das mesmas espécies, utilizando a extração por headspace, previamente otimizada, seguida de análise por GC-MS. Também se propôs a investigar a atividade antifúngica dos óleos essenciais de P. graveolens de diferentes origens e desenvolver uma formulação contendo uma nanoemulsão do óleo para o tratamento de candidíase vaginal. Por fim, teve como objetivo desenvolver metodologia de screening para avaliar a presença de DMAA e outros estimulantes em suplementos alimentares apreendidos, através de DART-MS/MS. Os resultados revelaram que a DMAA não está presente nos óleos essenciais de diferentes espécies de Pelargonium spp. obtidos por hidrodestilação, do Rio Grande do Sul. Após a otimização através de desenho experimental, a técnica de headspace provou ser eficaz na extração dos constituintes voláteis presentes nas folhas e, no entanto, a DMAA não foi detectada, assim como nos óleos essenciais comerciais de P. graveolens do Brasil, China, Egito, África do Sul, Albânia e Ilhas Reunião. Os óleos essenciais apresentaram atividade antifúngica frente às cinco espécies de Candida. Ainda, este efeito antifúngico apresentou melhores resultados com a nanoformulação contendo o óleo essencial. A análise de screening por DART-MS/MS se mostrou eficaz na detecção de DMAA, efedrina, sinefrina, cafeína, sibutramina e metilfenidato, em amostras de suplementos alimentares apreendidos, apresentando resultados positivos para todos os estimulantes. Com base nos resultados obtidos e nos objetivos propostos, verificou-se que mesmo após a 12 utilização de três técnicas analíticas distintas e uma nova alternativa para extração dos constituintes voláteis, a DMAA não foi econtrada nos óleos essenciais e nas folhas das espécies de Pelargonium, corroborando com outros estudos realizados, e indicando que a sua origem não é natural nestas espécies. A formulação final contendo a nanoemulsão com o óleo essencial apresentou atividade antifúngica superior a do óleo essencial livre. As análises das amostras apreendidas mostraram que mesmo após a sua proibição pelas agências regulatórias, os suplementos contendo DMAA e outros estimulantes ainda são comercializados, representando um grande risco para a saúde dos seus usuários. / 1,3-dimethylamylamine (DMAA) is a stimulant that started to be added in dietary supplements and weight loss compounds since 2006 and is widely consumed by athletes and the USA army. However, after reports of toxicity DMAA has been banned by regulatory agencies in Brazil and United States. However, even after its prohibition, DMAA still can be found in dietary supplements. Its origin was related to the essential oils of Pelargonium graveolens, and, however, many authors questioned the results and its natural origin. In addition, the essential oils of species of Pelargonium, had their antimicrobial activity reported against bacteria and fungi. Considering the aspects mentioned, this work aimed to determine the presence of DMAA in the essential oils by GC-MS, DART-MS/MS and LC-MS/MS; as well as in the leaves of the same species using the headspace extraction, previously optimized, followed by analysis through GC-MS. It has also been proposed to investigate the antifungal activity of essential oils of P. graveolens from different origins and develop a formulation containing an oil nanoemulsion for the treatment of vaginal candidiasis. Finally, it aimed to develop a screening method to evaluate the presence of DMAA and other stimulants in seized dietary supplements by DART-MS/MS. The results showed that DMAA is not present in the Rio Grande do Sul’s essential oils of Pelargonium spp. obtained by hydrodistillation. After optimization through experimental design, the headspace technique proved to be effective in extracting volatile constituents present in the leaves and, however, DMAA was not detected, as well as in commercial essential oils of P. graveolens from Brazil, China, Egypt, South Africa, Albania and Reunion Islands. The essential oils presented antifungal activity against five Candida species. Furthermore, this antifungal effect presented better results with the nanoformulation containing essential oil. DART-MS/MS screening was effective in detection of DMAA, ephedrine, synephrine, caffeine, sibutramine and methylphenidate in seized dietary supplements, showing positive results for all stimulants. Based on the results obtained and proposed objectives, it was verified that even after using three different analytical techniques and a new alternative for volatile constituents extraction, DMAA was not found in essential oils and leaves of Pelargonium spp., corroborating with other studies carried out, and indicating that its origin is not natural in these species. The final formulation containing the nanoemulsion with the essential oil had antifungal activity superior compared to 14 dispersed essential oil. The analysis of seized samples showed that even after its prohibition by regulatory agencies, supplements containing DMAA and other stimulants are still commercialized, representing a major health risk for their users.

Interés del estudio: El virus de la rotura del color de la flor del Pelargonium (Pelargonium flower break virus, PFBV) (género Carmovirus, familia Tombusviridae) constituye uno de patógenos virales más frecuentes en geranio, una planta ornamental de gran interés para la industria floral. En los últimos años se ha registrado un aumento progresivo de las infecciones causadas por este patógeno, con un porcentaje de incidencia superior al 80% en regiones de Europa occidental y en algunas áreas de España. La información acerca de las relaciones estructura-función en las proteínas de este virus era prácticamente inexistente al inicio de este trabajo y bastante limitada para las proteínas equivalentes de muchos virus relacionados, de modo que la aportación de datos adicionales en este campo puede ser de gran utilidad para entender mejor el establecimiento de la infección y los mecanismos de interacción con el huésped de este virus y de otros equivalentes. Objetivos: Durante el desarrollo de esta Tesis Doctoral se han llevado a cabo análisis de las relaciones estructura-función de las proteínas que codifica el PFBV, centrándonos en aquellos productos virales con características atípicas o cuya función no está del todo clara dentro del grupo al cual pertenece el virus. Con ello se ha pretendido aportar nuevos datos al conocimiento del patógeno y, en general, del grupo al que pertenece. Elementos de la metodología a destacar: La realización de esta Tesis ha permitido la publicación de varios artículos de divulgación científica, aprobados por comités que han avalado convenientemente el uso de las metodologías empleadas. Resultados logrados: Con este trabajo se ha establecido la etapa del ciclo infeccioso en la que se encuentran involucradas las distintas proteínas del PFBV y se han analizado relaciones estructura-función en dichos productos. Asimismo, se han determinado algunas propiedades relevantes de proteínas implicadas en la replicación, así como de aquellas que asisten / Martínez Turiño, S. (2012). ANÁLISIS FUNCIONAL DE PROTEÍNAS CODIFICADAS POR EL VIRUS DE LA ROTURA DEL COLOR DE LA FLOR DEL PELARGONIUM [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/16807 / Palancia

Thesis (MSc. (Horticulture)) -- University of Limpopo, 2016. / Pelargonium sidoides has numerous medicinal applications, with economic potential to serve as a future cultigen in smallholder farming systems. However, it is highly susceptible to the root-knot (Meloidogyne species) nematodes, without any identifiable nematode resistant genotypes. Nemarioc-AL and Nemafric-BL phytonematicides, with cucurbitacin A and cucurbitacin B active ingredients, respectively, are being researched and developed as an alternative to synthetic nematicides at the University of Limpopo. However, since active ingredients in phytonematicides are allelochemicals, the two phytonematicides have the potential of inducing phytotoxicity on crops protected against nematode damage. The objectives of the study, therefore, were (1) to determine the non-phytotoxic concentration of Nemarioc-AL phytonematicide on plant growth of P. sidoides, and (2) to determine the non-phytotoxic concentration of Nemafric-BL phytonematicide in plant growth of P. sidoides. Cuttings were raised in 30-cm-diameter plastic pots containing 10 000 ml steam-pasteurised river sand and Hygromix-T at 3:1 (v/v) under microplot conditions in autumn (March-May) and repeated in spring (August October) 2015. After establishment each plant was inoculated with 5 000 eggs and second-stage juveniles (J2s) of M. javanica. Six treatments, namely, 0, 2, 4, 6, 8 and 10% concentrations of each phytonematicide on separate trials were arranged in a randomised complete block design, with seven replicates. At 56 days after inoculation, in Experiment 1, Nemarioc-AL phytonematicide, treatment significantly (P ≤ 0.05) affected plant height, dry root mass and root galls, contributing 62, 69 and 70% to total treatment variation of the three variables, respectively. Relative to untreated control Nemarioc-AL phytonematicide increased plant height and dry root mass by 34 to 61% xxi and 20 to 76%, respectively, with a slight decrease by 5% in plant height at the highest concentration. However, the material decreased root galls by 5 to 50%. Significant (P ≤ 0.05) plant variables were subjected to Curve fitting-allelochemical respond dosage model, to generate biological indices which were used to compute the mean concentration stimulation point (MCSP) using the relation: MCSP = Dm + Rh/2 and the overall sensitivity value (∑k). In Experiment 1, MCSP = 6.18% and ∑k = 3. Plant variables and increasing concentration of phytonematicide exhibited quadratic relations. Treatments reduced nematode variables, at all levels including at the lowest, but the effect were not different. In Experiment 2, Nemarioc-AL phytonematicide treatment effects were not significant on plant variables except for root galls, but were significant for root nematodes except for eggs. Data for plant variables in Experiment 2 were not subjected to Curve fitting-allelochemical respond dosage model because they were not significant (P ≤ 0.05). In Experiment 1, Nemafric-BL phytonematicide treatment significantly (P ≤ 0.05) affected plant height and root galls, contributing 63 and 67% to total treatment variation of the two variables, respectively. Relatively to untreated control, plant height was increased by 10 to 36%, while root galls was reduced by 2.43 to 60%. In Experiment 1, MCSP = 2.87% and ∑k = 3. Concentrations of Nemafric-BL phytonematicide significantly (P ≤ 0.05) reduced eggs, juveniles and Pf at all levels including at the lowest, but the effect were not significant different, with treatments contributing 78, 72 and 90% to the total treatment variation. In Experiment 2, Nemafric BL phytonematicide treatment effects were not significant on plant variables except for root galls, but were significant for root. In conclusion, Nemarioc-AL and Nemafric-BL xxii phytonematicides could be applied at the lowest concentration of 2% where it was shown to be effective in suppressing population densities of M. javanica. / Agricultural Research Council (ARC), National Research Fund (NRF) , Flemish Inter university Council of Belgium and Land Bank Chair of Agriculture ─ University of Limpopo

No description available.

No description available.

Les huiles essentielles de citronnelle (Cymbopogon citratus), de géranium (Pelargonium graveolens) et de vétiver (Vetiveria zizanioides) sont utilisées partout dans le monde pour leur activité répulsive contre les principaux vecteurs (moustiques, tiques) de maladies infectieuses chez l'Homme (paludisme, chikungunya, …). L'application cutanée de ces produits naturels pour éviter le contact avec un vecteur n'avait pas été encore envisagée comme moyen de limiter les premiers stades de l’infection par l'agent pathogène transmis par le vecteur. Pour vérifier cette hypothèse, les travaux ont été consacrés à la mise en place d'un cadre structuré pour la réévaluation chimique et biologique des trois huiles essentielles sur le modèle du virus du Ross River (alphavirus) de la même famille que le virus du Chikungunya. La caractérisation chimique des huiles essentielles avec une technique de haute résolution (GC×GC/TOF-MS) a permis d'établir leur profil chémotypique précis. L'utilisation de marqueurs spécifiques (clones moléculaires du virus) a permis d'établir l'inhibition de la réplication virale en fonction des conditions d'application des huiles essentielles de géranium et citronnelle. Ces résultats suggèrent l'intérêt d’une huile essentielle répulsive dans les premiers stades d'une infection par un vecteur. À ce titre, l'étude comparative établit la haute valeur ajoutée de l'huile essentielle de géranium et oriente la recherche de nouveaux anti-infectieux naturels vers des complexes riches en monoterpènes. / Essential oils of citronella (Cymbopogon citratus), geranium (Pelargonium graveolens) and vetiver (Vetiveria zizanioides) are used worldwide as topical repellent against the main vectors (mosquitoes, ticks) of human infectious diseases (Malaria, chikungunya, …). Skin treatment with these natural products, initially to avoid contact with the vector had not yet been considered as a way to disrupt the early stages of infection when the repelling action fails. To check this hypothesis, a structured framework has been performed for the chemical and biological re-evaluation of the three essential oils. The latter was tested against Ross River virus (alphavirus) that belongs to the same family of Chikungunya virus. Analysis of essential oils using a high-resolution technique (GC × GC / TOF-MS) resulted in a more accurate chemotypical profile of the local production. The use of specific markers (molecular clones of the virus, Saclay CEA) allowed to establish the inhibition of viral replication depending of the conditions of geranium and citronella essential oils application. These results suggest the great interest of an essential oil topical repellent in the early stages of a vector infection. The comparative study established the high value of geranium essential oil and gave future direction to the discovery of new anti-infectious solutions from monoterpenes-rich natural complexes.