Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-21T13:55:26Z No. of bitstreams: 1 Cicero Nicolini.pdf: 1169299 bytes, checksum: e932282294084c1947939ed928971bcb (MD5) / Made available in DSpace on 2017-02-21T13:55:26Z (GMT). No. of bitstreams: 1 Cicero Nicolini.pdf: 1169299 bytes, checksum: e932282294084c1947939ed928971bcb (MD5) Previous issue date: 2011-09-01 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Brazil is the world’s largest passion fruit producer. The sanitary problems, mainly diseases caused by viruses are among the limiting factors for increasing productivity and longevity of the plantations. In Brazil, passion fruit woodiness, caused by Cowpea aphid-borne mosaic virus (CABMV) is the major viral disease of this crop. CABMV is also causing agent of the potyvirus mosaic, a viral disease that produces serious damages in cowpea. This crop in Brazil is important mainly in the Northeastern region and represents a relevant source of protein and starch for the population. Some years ago, in the state of Pernambuco it was found a potyvirus naturally infecting “lava-pratos”, leguminous plant native to Atlantic Forest commonly found on the margins of passionflower and cowpea crops. In the city of Paudalho it was found a group of “lava-pratos” plants presenting very intense symptoms with yellow spot on the leaves. From those plants, preparations examined in a transmission electron microscope showed the presence of elongated flexuous particles and cylindrical inclusions typical of potyvirus as well as isometric particles in high concentration and vesicles on the periphery of the chloroplasts which suggested the occurrence of a tymovirus. Serological analyses and coat protein sequencing to identify the potyvirus as well as studies on host adaptation in passionflower and cowpea, using isolates from these crops were undertaken. The virus with isometric particles was chemically purified to produce polyclonal antiserum and subjected to serological studies, host range and genome sequencing for characterization. It was concluded that the potyvirus found in “lava-pratos” is CABMV and phylogenetically observed the host adaptation of this virus to the families Fabaceae and Passifloraceae. For some strains geographical distance was not the strongest factor for genetic grouping and the Brazilian isolates form a large group distinct to other countries. According to the criteria of taxonomy, using amino acid sequence of coat protein and conserved region "tymobox", it was determined the presence of a tymovirus in “lava-pratos”, for which was proposed to be a new species named Cassia yellow mosaic-associated virus (CAYMaV). / O Brasil é o maior produtor mundial de maracujá. Os problemas fitossanitários especialmente as doenças ocasionadas por vírus estão entre os fatores limitantes ao aumento de produtividade e longevidade dos plantios. A principal virose do maracujazeiro no Brasil é o endurecimento dos frutos causado por Cowpea aphid-borne mosaic virus (CABMV). O CABMV também é o agente do mosaico de potyvírus em feijão-caupi, virose que provoca sérios danos nessa cultura. No Brasil, o feijão-caupi é produzido principalmente na Região Nordeste e representa fonte importante de proteína e amido para a população. Há alguns anos atrás, um potyvírus foi encontrado em Pernambuco infectando naturalmente o lava-pratos, uma leguminosa nativa da Mata Atlântica comumente encontrada nas bordas de cultivos de maracujá e feijão-caupi. No município de Paudalho foi observado um grupo de plantas de lava-pratos apresentando sintomas muito intensos com manchas amarelas nas folhas. Dessas plantas, ao ser analisadas preparações em microscópio eletrônico de transmissão, além da presença de partículas alongadas flexuosas e de inclusões cilíndricas típicas de potyvírus foram detectadas partículas isométricas em alta concentração e vesículas nas bordas dos cloroplastos, o que sugeria ser de um tymovírus. Foram realizadas análises sorológicas e seqüenciamento da capa protéica para identificação do potyvírus, assim como estudar a adaptação hospedeira a maracujazeiro e feijão-caupi, utilizando-se isolados obtidos dessas culturas. O vírus com partículas isométricas foi purificado quimicamente para produção de antissoro policlonal e submetido a estudos sorológicos, gama de hospedeiros e seqüenciamento de parte do genoma para caracterização. Concluiu-se que o potyvírus encontrado em lava-pratos é o CABMV e filogeneticamente se observou que ocorre adaptação hospedeira deste vírus em plantas das famílias Fabaceae e Passifloraceae. Para alguns isolados, a distância geográfica não foi o fator mais forte para agrupamento genético e os isolados brasileiros formam um grande grupo distinto aos de outros países. De acordo com os critérios de taxonomia, usando-se seqüência de aminoácidos da capa protéica e região conservada “tymobox”, foi determinada a presença em lava-pratos de um tymovírus, para o qual foi proposto ser uma nova espécie nomeada Cassia yellow mosaic-associated virus (CAYMaV).

This diploma thesis deals with the optimization of processes for extraction of bioactive lipophilic compounds from fruits of sea buckthorn (Hippophae Rhamnoides) into various types of plant oil bases using simple maceration. The theoretical part of this thesis deals with the characterization of this herb, its botanical classification, traditional use, chemical composition and medicinal effects. Increased attention is also paid to the characterization of individual types of plant oils used, namely camellia, camellia organic, passionflower, kukui and kiwi oil. The experimental part of the thesis deals with application of theoretical knowledge. The profile of total and free fatty acids for individual plant oil bases was determined by GC/FID, furthet the basic fat numbers were also determined. When optimizing the extraction, emphasis was placed not only on the effect of the extraction agent used, but also on the extraction time (1, 3, 5, 7, 10, 14, 21 and 66 days). The macerates were continuously subjected to the determination of selected parameters (total amount of carotenoids, total amount of phytosterols, lutein, neoxanthin, astaxanthin, stigmasterol, -sitosterol and vitamin E) using UV-VIS spectroscopy and HPLC/PDA. Likewise, the peroxide number was monitored during maceration to assess the degree of oxidative degradation of macerates. The recovery of selected total parameters in individual oils did not differ significantly in most cases. On the contrary, the yield of individual monitored parameters differed significantly. At the same time, static maceration under our conditions was not very suitable for the extraction of vitamin E, stigmasterol and total phytosterols.