Vacková, L.: Alkaloids Narcissus 'Dutch Master' (Amaryllidaceae) and their biological activity. I. Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2016. From a selected fraction ND-6, which was obtained by column chromatography of an alkaloid extract of Narcissus 'Dutch Master' (preparation of the alkaloid extract and column chromatography was performed by Mgr. Daniela Hulcová within her doctoral thesis), lycorine alakloid O-acetylpluviin was isolated using preparative TLC. Its structure was determined on the basis of MS, NMR analysis, and optical rotation, the obtained data were compared with the literature. The isolated alkaloid was tested on its possibility to inhibit human acetylcholinesterase and butyrylcholinesterase. The activity was expressed as IC 50 values (IC50 AChE = 648.03 ± 53.95 μM, IC50 BChE = 602.50 ± 48.50 μM) and compared with IC50 values of galanthamine, huperzine A and physostigmine. O-acetylpluviine showed a very low inhibitory cholinesterase activity, and so, the alkaloid does not seem to be a suitable cholinesterase inhibitor for potential use in the treatment of Alzheimer's disease. Keywords: Narcissus 'Dutch Master', Amaryllidaceae, lycorine alkaloids, Alzheimer's disease,...

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-01-26T13:50:02Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_CorrelacaoMetalobicaFungos.pdf: 2029628 bytes, checksum: 8feab0d533f4d04b3afb09538d403b20 (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-01-27T13:25:47Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_CorrelacaoMetalobicaFungos.pdf: 2029628 bytes, checksum: 8feab0d533f4d04b3afb09538d403b20 (MD5) / Made available in DSpace on 2017-01-27T13:25:47Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_CorrelacaoMetalobicaFungos.pdf: 2029628 bytes, checksum: 8feab0d533f4d04b3afb09538d403b20 (MD5) Previous issue date: 2016-06-30 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Os fungos endofíticos são uma fonte promissora de metabólitos secundários com aplicações biotecnológicas. Esses micro-organismos habitam o interior dos tecidos vegetais sem causar nenhum dano ao hospedeiro e como fruto dessa interação podem produzir algumas das substâncias sintetizadas pelas plantas hospedeiras. Neste contexto o uso de fungos endofíticos como fonte de biomoléculas em substituição as de planta representa vantagens econômicas e ambientais. Espécies da família Amaryllidaceae produzem alcaloides e outros metabólitos com atividades biológicas. Entre estas espécies destacam-se Crinum americanum L. E Hymenocallis littoralis (Jacq.) Salisb. No entanto, não há dados sobre fungos endofíticos de espécies da família. Diante dos exposto o objetivo deste trabalho foi investigar o potencial biotecnológico de fungos endofíticos de C. americanum e H. littoralis em comparação com as plantas hospedeiras e estudar a interação entre endofíticos e a planta hospedeira. Como resultados foram isolados 94 fungos endofíticos das duas espécies de Amaryllidaceae investigadas, dos quais 49 foram identificados e pertencem aos gêneros Colettotrichum, Acremonium, Trichoderma e Fusarium. Constatou-se que do total das linhagens analisadas 56 apresentaram lipídios em seus extratos, 21 cumarinas, 29 anronas e 2 apresentaram alcaloides. Foram selecionadas 12 linhagens de fungos endofíticos que apresentaram os melhores resultados na detecção de classes de metabólitos e os extratos foram analisados por Cromatografia em Camada Delgada e por Cromatografia Gasosa acoplada a Espectrometria de Massas em comparação com os extratos metanólicos das plantas hospedeiras. Constatou-se a correlação ente as classes de metabólitos detectados nos extratos dos fungos endofíticos e das plantas hospedeiras, indicando que estes micro-organismos são capazes de produzir algumas das mesmas substâncias que as plantas hospedeiras e a avaliação da atividade antimicrobiana destacou alguns extratos com atividade contra Candida parapsilosis. O extrato metanólico das folhas de H. littoralis e do fungo endofítico MIBA 0796 apresentaram os resultados mais relevantes com percentual de inibição acima de 60% contra a levedura. Com o estudo da interação entre fungos endofíticos de C. americanum e a planta hospedeira pode-se constatar que os micro-organismos ocorrem no interior das células vegetais e em condições favoráveis se desenvolveram e ocuparam também os espaços intercelulares. Esta localização dos endofíticos nos tecidos vegetais pode facilitar a troca de material genético entre o vegetal e os micro-organismos o que explicaria a correlação metabólica constatada nesta pesquisa. / The endophytic fungi are a promising source of secondary metabolites with biotechnological applications. These microorganisms inhabit the interior of plant tissues without causing any damage to the host and as a result of this interaction can produce some of the substances synthesized by the host plant. In this context the use of endophytic fungi as a source of biomolecules in the replacement of plant represents economic and environmental advantages. Species in the Amaryllidaceae family produce alkaloids and other metabolites with biological activities. Among these species include Crinum americanum L. and Hymenocallis littoralis (Jacq.) Salisb. However, no data on endophytic fungi from species of the family were found. In the view of the above stated the aim of this work was to scrutinize the biotechnological potential of endophytes of C. americanum and H. littoralis as compared to host plants and to study the interactions between endophytes and host plant. As results 94 endophytic fungi were isolated from two species of the investigated Amaryllidaceae, of which 49 were identified and belonging to the Colettotrichum class, Acremonium, Trichoderma and Fusarium. It was noted that of the total strains tested 56 showed lipids in its extracts, 21coumarins, 29 anthrones and 2 showed alkaloids. 12 strains of endophytic fungi were selected which presented the best results in the detection of classes of metabolites and the extracts were analyzed by Thin-layer chromatography and by Gas chromatography-mass spectrometry in comparison with the methanolic extracts of host plants. It was noted the correlation between the classes of metabolites detected in extracts of the endophytic fungi and host plants, indicating that these micro-organisms are able to produce some of the same substances that the host plants and the assessment of antimicrobial activity highlighted some extracts with activity against Candida parapsilosis. The methanolic extract of leaves of H. littoralis and endophytic fungus MIBA 0796 presented the most relevant outcomes with percentage of inhibition above 60% against yeast. With the study of the interaction between C. americanum endophytes and the host plant it can observe that microorganisms act within the plant cells and in favorable conditions develop and occupy intercellular spaces as well. This location of the Endophytes in plant tissues may facilitate the exchange of genetic material between plants and microorganisms explaining how metabolic correlation found in this research.

Many South African medicinal plants face the threat of over-collection for use in traditional medicines. Many bulbous plants suffer as the whole plant is removed from the wild so that the bulb may be used for medicine. Micropropagation is a technique which can be used as an alternative to conventional propagation methods. Micropropagation produces many plantlets in a relatively short period of time. Different plant parts of Brunsvigia undulata F.M. Leight, a rare South African species of medicinal value, were used in an attempt to produce in vitro plantlets using micropropagation techniques. Although leaf and floral explants were successfully formed from seedling explants and twin-scales. Seeds germinated quickly in culture. Seedlings which grew from seeds were cut into sections and used to initiate bulblets. Seedling explants formed bulblets, shoots and callus best when the explants included a meristematic region. Callus from seedling explants formed shoot clusters readily when placed on hormone-free MURASHIGE and SKOOG (1962) (MS) medium. Shoots from shoot clusters formed bulblets and rooted on medium supplemented with IBA. The greatest rooting response was achieved by bulblets on 1 mgl-1 IBA. The callus which was left after shoot clusters were separated was placed back onto hormone-free MS medium. Callus explants continued to form shoot clusters. Twin-scales, cut from large parent bulbs, were cultured on 25 hormone treatments. Bulblets formed on twin-scales even in the absence of plant growth hormones. Bulblets formed by twin-scales were used to determine the effects of both medium constituents and environmental factors on bulblet multiplication. Bulblet multiplication was greatest when bulblets were split in half and cultured as half-bulblets. Optimal multiplication was achieved on hormone-free MS, with 4% sucrose, kept at high temperatures in the dark. Bulblets were successfully initiated and multiplied from both seedlings and twin-scales. Bulblets which were produced via both protocols were acclimatized relatively easily. Both explant types could be used to mass propagate Brunsvigia undulata. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Please read the abstract in the section 07chapter7 of this document / Thesis (DPhil (Plant Physiology))--University of Pretoria, 2007. / Plant Science / unrestricted

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