Graduation date: 1992

Graduation date: 2004

Cuphea (Lythraceae) is a genus of interest as a possible domestic source of medium-chain fatty acids. The genus is in the preliminary phases of domestication. An unsuccessful attempt was made to produce interspecific hybrids between species in several sections of the genus and with varying chromosome numbers. An analysis was made of the time of day and size bud to collect for cytological study of meiotic phases. Chromosome numbers were reported for C. ignea and C. pinetorum. Arm-pairing data were collected for 4 species and one interspecific hybrid. Metaphase I bivalent size was measured for 4 species. A genetic study was done in C. procumbens. Hypotheses regarding the inheritance of flower color, viscid plant hairs, seed coat spots, seed coat hairs and seed dormancy are proposed. A PEG osmoticum pre-germination treatment was found ineffective in breaking dormancy.

Cuphea is an herbaceous genus having species whose seed storage lipids are predominantly medium-chain triglycerides (MCTs). Cuphea lanceolate Ait, and Cuphea viscosissima Jacq. are central to the breeding of Cuphea as a new commercial source of MCTs. The objectives of this thesis were to develop a reliable method to extract DNA from Cuphea, to construct a molecular-marker linkage map of C. lanceolate, and to evaluate quantitative-trait loci (QTL) affecting seed capric acid content, seed oil content, and embryo development in C. lanceolate. We used allozyme and restriction-fragment- length-polymorphism (RFLP) markers to study Cuphea because molecular markers are the best method available to investigate the Mendelian genetics underlying quantitative traits, and are useful in breeding. Adequate yield of clean DNA is essential for RFLP mapping. We tried numerous DNA extraction methods that failed to remove contaminants that interfere with restriction digests of Cuphea DNA. The method described here was developed to remove those contaminants and maintain relatively high DNA yields. In this method, the critical step in purification consists of washing the DNA with phenol while it is complexed with CTAB and dissolved in 1 M NaCl. An RFLP and allozyme linkage map of C. lanceolate was constructed having 37 markers in six linkage groups with a total distance of 288 cM. Levels of polymorphism were estimated for three lines of C. lanceolate and one line of C. viscosissima using 84 random genomic clones and two restriction enzymes, EcoRI and HindIII. Twenty-nine percent of the probes detected RFLPs between C. lanceolate lines, whereas 63% of the probes detected RFLPs between C. lanceolate and C. viscosissima lines. Thirty RFLP and four allozyme markers were used to locate on the C. lanceolata linkage map QTL affecting seed capric acid content, seed oil content, and embryo development. Three unlinked QTL explained 19.4% of the phenotypic variation in capric acid content in F₂ seed. Seed oil content and seed weight were measured on seed from field-grown F₂ plants. Seed weight was indicative of embryo size (development). Four unlinked QTL explained 33.9% of the phenotypic variation in embryo size. One of these QTL, which explained 20.3% of the variation, may have been a chromosomal deletion detected by a marker having a null allele. Three QTL pleiotropically affected seed oil by affecting embryo size. The one QTL that only affected seed oil accounted for 2.8% of the phenotypic variation. / Graduation date: 1991

Graduation date: 1992

Graduation date: 1997

No description available.

Submitted by Sandro Camargo (sandro.camargo@unipampa.edu.br) on 2015-05-08T02:43:19Z No. of bitstreams: 1 127110045.pdf: 1527824 bytes, checksum: 960729a0a23069d5d40ec997cc3034b8 (MD5) / Made available in DSpace on 2015-05-08T02:43:19Z (GMT). No. of bitstreams: 1 127110045.pdf: 1527824 bytes, checksum: 960729a0a23069d5d40ec997cc3034b8 (MD5) Previous issue date: 2014-07-17 / O gênero Cuphea, popularmente conhecido no Brasil por “sete-sangrias”, tem seu uso medicinal reconhecido devido aos efeitos diurético, hipotensor e cardioprotetor. No sul do Brasil, em região característica do bioma Pampa, foi encontrada a espécie Cuphea glutinosa Cham. & Schltdl. Embora o uso popular, esta espécie é pouco descrita na literatura. O presente trabalho tem como objetivos o estudo da composição química dos extratos de C. glutinosa e a avaliação das atividades antifúngica e anti-hipertensiva. O material vegetal foi coletado na cidade de Uruguaiana (RS, Brasil), identificado e depositado em herbário. Após secagem e trituração do material vegetal, foram obtidos os extratos hidroetanólicos através de maceração exaustiva com etanol 40% (v/v) para folhas e etanol 70% (v/v) para raízes. Para a infusão, utilizou-se água a 80oC. As análises cromatográficas foram realizadas em equipamento cromatógrafo a líquido Prominence Shimadzu, em técnica por CLAE e CLUE. Utilizou-se sistema de fase reversa, eluição por gradiente com fase móvel composta por acetonitrila:metanol (4:1) e ácido fórmico 0,1% pH 3,0, coluna C18 analítica e fast, e detecção por UV-DAD e ESI-MS. Os teores de polifenóis totais e de flavonóides foram determinados por método colorimétrico, seguindo metodologia padronizada. A atividade antifúngica in vitro foi realizada utilizando o método de microdiluição em caldo, determinando-se a CIM, in-vitro, contra diferentes isolados clínicos. Para avaliação do potencial anti-hipertensivo in vivo, foram realizadas medições da pressão sanguínea pelo método de monitoramento hemodinâmico invasivo, através da inserção de cateter na artéria carótida. Os resultados de teor de fenólicos totais indicaram predominância destes componentes em extratos obtidos de folhas e por maceração, conforme os valores obtidos: 1,8501 mg EAG/mL (folhas) e 0,8467 mg EAG/mL (raízes) para infusão, e 3,7284 mgEAG/mL (folhas) e 2,6266 mg EAG/mL (raízes) para maceração. Quanto ao teor de flavonóides, os resultados quantitativos foram: 7,0959 mg/g (folhas) e 0,5664 mg/g (raízes) para a infusão, e 7,9511 mg/g (folhas) e 0,5994 mg/g (raízes) para maceração. Na análise cromatográfica, os extratos obtidos das folhas de C. glutinosa apresentaram picos cromatográficos bem separados, em perfil reprodutível. Na determinação por CLUE-MS, os dados de íon molecular e fragmentos de massa indicaram a composição predominante em flavonóides, sugerindo-se os componentes quercetina-3-O-glicosídeo, quercetina-3- arabinosídeo, quercetina-3-glicuronídeo, isoramnetina e quercetina-5-O-β-glicopiranosídeo. Para o potencial antifúngico, os extratos das folhas e raízes apresentaram atividade in vitro contra Candida parapsilosis, Candida tropicalis e Trichosporon asahii, com CIM variando na faixa de 1,9-62,5 μg/mL. Nos testes hemodinâmicos realizados, os extratos das folhas não apresentaram efeito significativo sobre a pressão arterial. A identificação dos componentes em C. glutinosa, derivados de quercetina, torna promissora novas investigações a fim de aprofundar o conhecimento a respeito desta espécie, em especial na busca de respostas para a relatada ação anti-hipertensiva. / The Cuphea genus, popularly known in Brazil as "sete-sangrias", is used traditionally due the diuretic, hypotensive and cardioprotective effects. In southern Brazil, in characteristic region of Pampa biome, it was found the species Cuphea glutinosa Cham. & Schltdl. Although used popularly, this species is few reported in the literature. The present work aimed to study the chemical composition of extracts from C. glutinosa and to evaluate the antifungal and anti -hypertensive activities. The plant material was collected in the city of Uruguaiana (RS, Brazil), identified and deposited in a herbarium. After dryness and milling, the hydroethanolic extracts were obtained through exhaustive maceration using ethanol 40% (v/v) for leaves and ethanol 70% (v/v) for roots. The infusions were prepared using water at 80 °C. The chromatographic analyses were performed in liquid chromatography Prominence Shimadzu, for HPLC and UPLC assays. The method was conducted using reverse phase system, gradient elution with mobile phase composed by acetonitrile:methanol (4:1) and formic acid 0.1% pH 3.0, C18 analytical and fast column, and detection by UV-DAD and MS. The polyphenols and flavonoids contents were determined by colorimetric method. The in vitro antifungal activity was conducted by using the broth microdilution method, determining the MIC against different clinical isolates. For evaluation of in vivo anti-hypertensive potential, the blood pressure was measured by the method of invasive hemodynamic monitoring, through of insertion the catheter into the carotid artery. The results of phenolic content indicated the high concentration of these compounds in leaves extracts obtained by maceration: 1.8501 mgEAG/mL (leaves) and 0.8467 mgEAG/mL (roots) for infusion, and 3.7284 mgEAG/mL (leaves) and 2.6266 mgEAG/mL (roots) for maceration. For flavonoids, the contents were: 7.0959 mg/g (leaves) and 0.5664 mg/g (roots) for infusion, and 7.9511 mg/g (leaves) and 0.5994mg/g (roots) for maceration. In the chromatographic analyses, the leaf extracts from C. glutinosa presented chromatographic peaks well separated and reproducible. In the determination by UPLC-MS, the molecular ion and mass fragments indicated the predominant composition in flavonoids, suggesting the compounds quercetin-3- O-glucoside, quercetin-3-arabinoside, quercetin-3-glucuronide, isorhamnetin and quercetin-5- O-β-glucopiranoside. For the antifungal potential, the leaf and roots extracts presented activity against Candida parapsilosis, Candida tropicalis e Trichosporon asahii, with MIC values ranging 1,9-62,5 μg/mL. In the hemodynamic tests performed, the leaves extracts did not present significant effect in the arterial pressure, although a tendency in pressure reduction could be observed. The identification of quercetin derivatives in C. glutinosa becomes promisor further investigations about this species, mainly in respect to the anti-hypertensive action.