The upper Paleozoic miospore genus Spelaeotriletes Neves and Owens, 1966, and constituent Gondwanan species

The upper Paleozoic miospore genus Spelaeotriletes Neves and Owens, 1966 is reviewed as a morpho-taxonomic entity and vis-a-vis other similarly constructed (pseudosaccate) genera - Geminospora Balme, 1962, Grandispora Hoffmeister, Staplin, and Malloy, 1955, Rhabdosporites Richardson, 1960, and Retispora Staplin, 1960. Detailed studies of numerous, mainly topotype specimens of Spelaeotriletes ybertii (Marques-Toigo, 1970) Playford and Powis, 1979 from the Lower Permian of Uruguay result in its re-diagnosis, in conjunction with a survey of its exclusively Gondwanan occurrences, particularly in South American strata extending from the Upper Carboniferous (Westphalian) into the Lower Permian, and also in Australian strata of approximately equivalent age. The characteristics of other species of Spelaeotriletes reported from upper Paleozoic deposits of Gondwana are discussed, as are their temporal representations in various broad regions of the supercontinent (South America, North Africa, Australia). These species include two, perhaps three, that, like Spelaeotriletes triangulus/arenaceus, are known also from Euramerica - S. balteatus (Playford, 1963) Higgs, 1996, S. pretiosus (Playford, 1964) Utting, 1987, and possibly S. owensii Loboziak and Alpern, 1978. Other species, such as S. benghaziensis Loboziak and Clayton, 1988, S. giganteus Loboziak and Clayton, 1988, and S. vibrissus Playford and Satterthwait, 1988, have, on present knowledge, exclusively Gondwanan occurrences. S. queenslandensis Jones and Truswell. 1992, known only from Upper Carboniferous strata of northeastern Australia, is formally reassigned on sculptural grounds to Grandispora. Not unexpectedly in a paleogeographic perspective, North Africa and South America are more closely allied with each other than with Australia in terms of shared species of Spelaeotriletes. (C) 2001 Elsevier Science Ltd. All rights reserved.

Stomatogenesis in the ciliate genus Macropodinium Dehority, 1996 (Litostomatea : Macropodiniidae)

Stomatogenesis and the cell division cycle was investigated for Macropodinium yalanbense Dehority, 1996 from Macropus giganteus using light and electron microscopy. Macropodinium spp. are endosymbiotic ciliates found only in the stomachs of macropodid marsupials. Stomatogenesis proceeds through 4 stages: initial formation of a transverse division suture; formation of the preoral field and formation of vestibular kineties in an internal pouch; extension of vestibulum posteriorly and external formation of new adoral kineties; and extension of somatic and adoral kineties accompanying dorsal and ventral constriction of the cell. Karyokinesis and formation of the new cytoproct occur immediately prior to cytokinesis. Comparison with other litostome ciliates shows that the formation of new vestibular kineties is most similar to that of the entodiniomorphs, formation of adoral kineties is most similar to that of the haptorians and formation of the somatic kineties to that of the vestibuliferans. The phylogenetic affinities of Macropodinium are thus difficult to infer from the ontogeny of organelle systems. Stomatogenesis of the adoral kineties is either epiapokinetal or a new type of cryptotelokinetal whereas the vestibular kineties are formed by either endoapokinetal or cryptotelokinetal processes. No other ciliate has been observed to utilise 2 types of stomatogenesis in its division cycle.

Potential of biologically active plant oils to control mosquito larvae (Culex pipiens, Diptera: Culicidae) from an egyptian locality

The insecticidal effect of six commercially available plant oils was tested against 4th larval instars of Culex pipiens. Larvae were originally collected from Meit El-Attar, Qalyubia Governorate, Egypt, and then reared in the laboratory until F1 generation. The LC50 values were 32.42, 47.17, 71.37, 83.36, 86.06, and 152.94 ppm for fenugreek (Trigonella foenum-grecum), earth almond (Cyperus esculentus), mustard (Brassica compestris), olibanum (Boswellia serrata), rocket (Eruca sativa), and parsley (Carum ptroselinum), respectively. The tested oils altered some biological aspects of C. pipiens, for instance, developmental periods, pupation rates, and adult emergences. The lowest concentrations of olibanum and fenugreek oils caused remarkable prolongation of larval and pupal durations. Data also showed that the increase of concentrations was directly proportional to reduction in pupation rates and adult emergences. Remarkable decrease in pupation rate was achieved by mustard oil at 1000 ppm. Adult emergence was suppressed by earth almond and fenugreek oils at 25 ppm. In addition, the tested plant oils exhibited various morphological abnormalities on larvae, pupae, and adult stages. Consequently, fenugreek was the most potent oil and the major cause of malformation of both larval and pupal stages. Potency of the applied plant oils provided an excellent potential for controlling C. pipiens.

Influência do manejo da cultura no balanço energético da utilização de Miscanthus na produção de energia, em Portugal

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Energia e Bioenergia

Fitorremediação de águas residuais contaminadas com Cd e Ni por três genótipos de Miscanthus

Fitorremediação, o uso de vegetação para a descontaminação de diferentes matrizes, é uma estratégia de baixo custo e tem sido apontada como uma metodologia passível de atenuar e estabilizar águas residuais. Neste contexto, este estudo tem como principal objectivo avaliar a capacidade fitorremediadora de três genótipos de Miscanthus (duas espécies - Miscanthus sinensis e Miscanthus floridulus, e um híbrido – Miscanthus x giganteus) a águas residuais contaminadas com cádmio e níquel (2,0 mg Ni/L e 0,2 mg Cd/L). Os resultados demonstraram que a irrigação com águas residuais contaminadas com Cd e Ni não afecta a produtividade dos três genótipos de Miscanthus em estudo e que o M. x giganteus e o M. floridulus registaram produtividades mais elevadas que o M. sinensis. Contudo, o M. x giganteus, demonstrou tendencialmente uma redução da sua produção com a contaminação de Cd. A biomassa obtida dos ensaios irrigados com águas residuais contaminadas apresentaram maior teor de Cd e Ni, do que a biomassa recolhida dos ensaios Controlo. O sistema Miscanthus-solo registou valores de remoção de Cd de 100%, e de Ni de 96-100%, no caso do M. x giganteus e do M. sinensis. O M. floridulus, contudo, demonstrou menos potencial de fitorremediação no que diz respeito à remoção de Ni das águas residuais: apenas 58% de Ni foi removido pelo sistema “biofiltrador”. Ainda assim, verificou-se a acumulação de Cd e Ni no perfil superior do solo devido à irrigação com as águas residuais contaminadas, o que demonstra a necessidade de controlo desta técnica de modo a prevenir possíveis efeitos negativos no ambiente.

Adaptação fenotípica de raízes de várias plantas com frutos de Dictyosperma rubra Wendl. & Drude encontrados no solo

Resumimos, neste trabalho, os resultados das observações sobre a adaptação fenotípica de raízes de várias plantas, tais como grama-batatais - Paspalum notatum, quebra-pedra ou erva-pombinha Phyllathus ninuri, maria-preta - Solanum nigrum, carurú - Amaranthus sp., tiririca - Cyperus rotundus e outras, com os frutos encontrados no solo da palmeira Dictyosperma rubra Wendl. & Drude, cultivada como ornamental no Parque da Escola Superior de Agricultura "Luiz de Queiroz". Essa adaptação caracteriza-se pela penetração das raízes através de orifícios circulares, existentes no pericarpo dos frutos, e produzidos por uma broca da família Bostriquide, da ordem Coleoptera. As raízes assim assemelham-se a um colar. Diante desse fato, decidimos montar um experimento que nos permitisse elucidar o mecanismo dessa curiosa adaptação fenotípica.

Ação alelopática de alguns extratos de plantas daninhas na germinação do arroz (Oryza sativa L. cv. IAC - 165)

O efeito do extrato aquoso de três plantas daninhas, Cynodon dactylon (l.)Pers., Cyperus rotundus l. e Sorghum halepense (l.) Pers. sobre a germinação e o crescimento de plântulas de arroz (Oryza salina L. cv. IAC-165) foi estudado, com a finalidade de investigar a sensibilidade do arroz aos efeitos alelopáticos dessas plantas. Foram utilizados, na preparação dos extratos, raizes de Cynodon daclylon, tubérculos de Cyperus rolundus: e riomas de Sorglun halepense, os quais foram homoneizados em solução aquosa, submetidos a filtração e centrifugação. O sobrenadante foi aplicado no substrato de germinação das sementes, verificando-se, aos sete dias após a se meadura, que apenas os extratos de Cy-nodon daetylon e Sorghum halepense afetaram a porcentagem de germinação das sementes de arroz. Entretanto, o crescimento da radícula e da parte aérea das plântulas foi prejudicado pelos três extratos testados, sendo o efeito mais drástico causado pelo extrato de Cyperus, retundus.

Taxonomia da subfamília Corinninae (Araneae, Corinnidae) nas regiões neotropical e neártica

The subfamily Corinninae is characterized and diagnosed. Two synapomorphies are hypothesized for the subfamily, both regarding the male palpal reservoir, which is primarily coiled and presents a sclerotized distal sector. Seventeen genera are recognized, six of which are new: Abapeba (type species Corinna lacertosa Simon), Erendira (type species Corinna pallidoguttata Simon), Septentrinna (type species Corinna bicalcarata Simon), Simonestus (type species Diestus validus Simon), Tapixaua (type species T. callida sp. nov.) and Tupirinna (type species T. rosae sp. nov.). The genera Creugas Thorell, Falconina Brignoli and Paradiestus Mello-Leitão are revalidated. Diestus Simon and Lausus Simon are newly synonymized with Corinna C. L. Koch. Chemmis Simon is included in the synonymy of Megalostrata Karsch. Hypsinotus L. Koch is removed from the synonymy of Corinna and included in the synonymy of Creugas. Thirteen new species are described: Septentrinna yucatan and S. potosi from Mexico; Tupirinna rosae from Venezuela and Brazil; Tapixaua callida from Brazil and Peru; Abapeba hoeferi, A. rioclaro, A. taruma, Corinna ducke, C. colombo, C. mourai, C. recurva and Parachemmis manauara from Brazil; Creugas lisei from Brazil, Argentina and Uruguay. Twenty seven species are redescribed. Fifty eight new combinations are presented: from Chemmis, Septentrinna steckleri (Gertsch); from Corinna, Abapeba abalosi (Mello-Leitão), A. cleonei (Petrunkevitch), A. echinus (Simon), A. grassima (Chickering), A. guanicae (Petrunkevitch), A. lacertosa (Simon), A. luctuosa (F. O. Pickard-Cambridge), A. lugubris (Schenkel), A. pennata (Caporiacco), A. kochi (Petrunkevitch), A. saga (F. O. Pickard-Cambridge), A. wheeleri (Petrunkevitch), Creugas annamae (Gertsch & Davis), C. apophysarius (Caporiacco), C. bajulus (Gertsch), C. bellator (L. Koch), C. bicuspis (F.O. Pickard-Cambridge), C. epicureanus (Chamberlin), C. falculus (F. O. Pickard-Cambridge), C. mucronatus (F. O. Pickard-Cambridge), C. navus (F. O. Pickard-Cambridge), C. nigricans (C. L. Koch), C. plumatus (L. Koch), C. praeceps (F. O. Pickard-Cambridge), C. silvaticus (Chickering), C. uncatus (F. O. Pickard-Cambridge), Erendira luteomaculatta (Petrunkevitch), E. pallidoguttata (Simon), E. subsignata (Simon), Falconina albomaculosa (Schmidt), F. crassipalpis (Chickering), F. gracilis (Keyserling), Megalostrata raptrix (L. Koch), Paradiestus egregius (Simon), P. giganteus (Karsch), P. penicillatus (Mello-Leitão), P. vitiosus (Keyserling), Septentrinna bicalcarata (Simon), S. paradoxa (F. O. Pickard-Cambridge), S. retusa (F. O. Pickard-Cambridge), Simonestus pseudobulbolus (Caporiacco), S. robustus (Chickering), S. semiluna (F.O. Pickard-Cambridge), Stethorrhagus maculatus (L. Koch) and Xeropigo smedigari (Caporiacco); from Diestus, Corinna alticeps (Keyserling), C. kochi (Simon), Simonestus occidentalis (Schenkel), S. separatus (Schmidt) and S. validus (Simon); from Lausus, Corinna grandis (Simon) and Abapeba sicarioides (Mello-Leitão); from Medmassa, Corinna andina (Simon) and C. venezuelica (Caporiacco); from Megalostrata, Erendira atrox (Caporiacco) and Erendira pictitorax (Caporiacco); from Parachemmis, Tupirinna trilineata (Chickering). Five combinations are restaured: Corinna aenea Simon, Creugas cinnamius Simon, Creugas gulosus Thorell, Falconina melloi (Schenkel), Paradiestus aurantiacus Mello-Leitão. Twenty five new synonymies are proposed: Diestus altifrons Mello-Leitão with Corinna nitens (Keyserling); Corinna tomentosa Simon, C. tridentina Mello-Leitão, Hypsinotus flavipes Keyserling, H. humilis Keyserling and Xeropigo scutulatus Simon with Xeropigo tridentiger (O. Pickard-Cambridge); Corinna cribosa Mello-Leitão and C. stigmatica Simon with Falconina gracilis (Keyserling); Corinna casueta Chickering with SIMONestus separatus (Schmidt); Corinna abnormis Petrunkevitch, C. antillana BRYANT, C. consobrina Simon, C. inornata Kraus, C. nervosa F. O. Pickard-Cambridge, C. wolleboeki Banks, Creugas cetratus Simon, C. senegalensis Simon and Hypsinotus gracilipes Keyserling with Creugas gulosus Thorell; Chemmis frederici Simon, Delozeugma formidabile O. Pickard-Cambridge, D. mordicans O. Pickard-Cambridge, Megalostrata sperata Kraus and M. venifica KARSCH with Megalostrata raptrix (L. Koch); Megalostrata lohmanderi Caporiacco with Erendira atrox (Caporiacco); Corinna tenubra Chickering with Parachemmis fuscus Chickering. One new name, Creugas berlandi, is erected for Corinna bellatrix Schmidt. Males of Creugas cinnamius, Corinna kochi, Methesis semirufa Simon, Paradiestus aurantiacus, Septentrinna steckleri and Xeropigo smedigari, the females of Paradiestus giganteus, Septentrinna bicalcarata and the adult female of S. steckleri are described for the first time.

Trypanorhynch Cestodes of Commercial Fishes from Northeast Brazilian Coastal Waters

A large scale investigation on trypanorhynch cestode infestation of tropical marine fishes was carried out along the Northeast Brazilian coast in the summer of 1991 and 1993. A total of 798 fish specimens belonging to 57 species and 30 families were examined. Metacestodes of 11 different trypanorhynchs were found: Callitetrarhynchus gracilis, Dasyrhynchus giganteus, Grillotia sp., Nybelinia edwinlintoni, N. indica, N. senegalensis, Nybelinia c.f. lingualis, Otobothrium cysticum, Pseudolacistorhynchus noodti, Pseudotobothrium dipsacum and Pterobothrium kingstoni. Scanning electron microscopy was used to clarify details of the tentacular armature of some species. Rose-thorn shaped hooklets, regularly arranged like microtriches, are described from the bothridial surface of N. edwinlintoni. Of the 57 fish species, 15 harboured trypanorhynch cestodes. Of these the mullid Pseudupeneus maculatus was the most heavily infested fish species, harbouring 5 different trypanorhynch species. P. noodti in P. maculatus had the highest prevalence (87%) and intensity (maximum = 63) of infestation. C. gracilis was the parasite with the lowest host-specificity. It could be isolated from 10 fish species. The cestode fauna of the Northeast Brazilian coast appears to be similar to that of the West African coast. Five of the trypanorhynch cestodes found during this study are common to both localities. The two single cases of intra musculature infestation in Citharichthys spilopterus and Haemulon aurolineatum by trypanorhynch cestodes indicate that marketability of the investigated commercially exploited fish species is inconsequential.

Bone anabolic potential of soy isoflavones and plant extracts and genomic changes during differentiation of hPOB-tert osteoblast-like cells

Summary For the nutritional management of bone health and the prevention of osteoporosis it is important to identify nutrients that positively influence the bone remodeling process at the cellular level. Soy isoflavones show promising osteoprotective effects in animals and humans but their mechanism of action in bone cells is yet poorly understood. Firstly, soy tissue cultures were characterized as a new and optimized source of isoflavones. A large variability in the isoflavone content was observed and high-producing strains (46.3 mg/g dry wt isoflavones) were identified. In the Ishikawa cells bioassay, the estrogenicity of isoflavones was confirmed to be 1000 to 10000 less than 17Mestradiol and that of the malonyl forms was shown for the first time (EC50 of 350 nM and 1880 nM for malonylgenistin and malonyldaidzin, respectively). The estrogenic activity of soya tissue culture extracts correlated to their isoflavone content. Secondly, the effects of phytonutrients on BMP-2 gene expression and on the mevalonate synthesis pathway, as key mediators of bone formation, were investigated. Dietary achievable concentrations of genistein and daidzein (10vM), and statins (4xM) but not 17M estradiol (10nM), induced BMP-2 gene expression (by up to 3-fold) and inhibited the cholesterol biosynthetic pathway (by up to 50%) in the human osteoblastic cell line hP0B¬tert. In addition, several plant extracts (Cyperus rotundus, Lindera benzoin and Cnidium monnieri) induced BMP-2 gene expression but this induction was not restricted to the inhibition of the cholesterol synthesis pathway neither to the estrogenicity. Finally, the gene expression profiles during hP0B-tert differentiation induced by vitamin D and dexamethasone were analyzed with the Affymetrix human GeneChip. 1665 different genes and 98 ESTs were significantly regulated. The expression profiles of bone-related genes was largely in agreement with previously documented patterns, supporting the physiological relevance of the genomic results and the hP0B-tert cell line as a valid model of human osteoblast differentiation. The expression of alternative differentiation markers during the osteogenic treatment of hP0B-tert cells indicated that the adipocyte and myoblast differentiation pathways were repressed, confirming that these culture conditions allowed only osteoblast differentiation. The gene ontology analysis identified further sub-groups of genes that may be involved in the bone formation process. Aims of the thesis In order to define new strategies for the nutritional management of bone health and for the prevention of osteoporosis the major goal of the present work was to investigate the potential of phytonutrients to positively modulate the bone formation process at the cellular level and, in particular: 1.To select and optimise alternative plant sources containing high levels of isoflavones with estrogenic activity (Chapter 3). 2.To compare the effects of statins and phytonutrients on BMP-2 gene expression and on the mevalonate synthesis pathway and to select new plant extracts with a bone anabolic potential (Chapter 4). 3.To further characterize the new human periosteal cell line, hP0B-tert, as a bone- formation model, by elucidating its gene expression profile during differentiation induced by vitamin D and dexamethasone (Chapter 5).

Relatório Preliminar sobre a Biodiversidade - Complexo de Árias protegidas do Leste da Ilha da Boa Vista

O Complexo de Áreas Protegidas do Leste da Ilha da Boa Vista (CAPLBV) inclui áreas terrestres, costeiras e marinhas bem como algumas colinas de baixa altitude em suas zonas terrestres (como a Ponta de Chã de Tarafe e o Monumento Natural de Monte Estância) na parte oriental da ilha da Boa Vista e estende-se por uma vasta área desde a Ponta de Ajudante a sul até a Ponta de Chã de Tarafe a norte. A biodiversidade da ilha da Boa Vista é caracterizada pela existência de várias comunidades de fauna e flora representativas dos ecossistemas costeiros e marinhos de Cabo Verde, da qual se destaca a tartaruga marinha Caretta caretta que aqui tem a sua principal área de desova em Cabo Verde. A vegetação costeira inclui Sporobolus spicatus, Cakile maritima, Sesuvium sesuvioides, Zygophylum fontanesii e Zygophylum simplex, sendo que as espécies mais representativas nas áreas lagunares são Arthrocnemum glaucum, Zygophylum waterlotii, Zygophylum fontanesii, Sporobolus minutus, Sporobolus spicatus e Cyperus bulbosus. A avifauna associada inclui Charadrius alexandrinus, Himantopus himantopus, Arenaria interpres, Pluvialis squatarola, Tringa nebularia, Ardea cinerea, Egretta garcetta, Ibis bulbucus, Platalea leucorodia, Pandion haliaetus, Fregata magnificens, Calonectris edwardsii, Sula leucogaster, Phaethon aethereus, Pelagodroma marina e Oceanodroma castro. As comunidades de corais ao longo da costa da ilha da Boa Vista, nomeadamente no ilhéu de Sal Rei e na baía das Gatas são das mais diversificadas e abundantes de todo o território de Cabo Verde (Cabo Verde 2000, 2001). As espécies do género Conus apresentam uma elevada diversidade e um elevado grau de endemismo. Várias espécies de tubarões e peixes pelágicos bem como mamíferos marinhos se reproduzem nas águas costeiras da Boa Vista. O presente relatório tem como objectivo caracterizar o ambiente terrestre, costeiro e marinho em geral, identificar e avaliar a situação da biodiversidade na área proposta para fazer parte do CAPLBV e nas áreas circundantes, através do levantamento, com recurso à pesquisa bibliográfica e saídas de campo, da diversidade e abundância da flora e fauna, em especial das espécies em vias de extinção e as endémicas, das espécies com importância ecológica no contexto internacional decorrente da posição biogeográfica do arquipélago e das espécies com importância ecológica e económica para a ilha da Boa Vista bem como para o arquipélago de Cabo Verde.

Soil solarization for weed control in carrot

Soil solarization is a technique used for weed and plant disease control in regions with high levels of solar radiation. The effect of solarization (0, 3, 6, and 9 weeks) upon weed populations, carrot (Daucus carota L. cv. Brasília) yield and nematode infestation in carrot roots was studied in São Luís (2º35' S; 44º10' W), MA, Brazil, using transparent polyethylene films (100 and 150 mm of thickness). The maximum temperature at 5 cm of depth was about 10ºC warmer in solarized soil than in control plots. In the study 20 weed types were recorded. Solarization reduced weed biomass and density in about 50% of weed species, including Cyperus spp., Chamaecrista nictans var. paraguariensis (Chod & Hassl.) Irwin & Barneby, Marsypianthes chamaedrys (Vahl) O. Kuntze, Mitracarpus sp., Mollugo verticillata L., Sebastiania corniculata M. Arg., and Spigelia anthelmia L. Approximately 40% of species in the weed flora were not affected by soil mulching. Furthermore, seed germination of Commelina benghalensis L. was increased by soil solarization. Marketable yield of carrots was greater in solarized soil than in the unsolarized one. It was concluded that solarization for nine weeks increases carrot yield and is effective for controlling more than half of the weed species recorded. Mulching was not effective for controlling root-knot nematodes in carrot.

Efeitos da solarização do solo na densidade populacional da tiririca e na produtividade de hortaliças sob manejo orgânico

Foi conduzido um ensaio de campo em Seropédica, Estado do Rio de Janeiro, para avaliar o efeito da solarização do solo na população infestante de tiririca (Cyperus rotundus) e na produtividade de hortaliças submetidas a manejo orgânico. A solarização correspondeu à cobertura do solo com polietileno transparente (50 mim) por um período de 210 dias. Houve um aumento da temperatura média da camada superficial do solo (0-10 cm) da ordem de 23%, em relação ao das parcelas não-solarizadas. Após a retirada do plástico, cultivaram-se: cenoura (Daucus carota 'Brasília'), repolho (Brassica oleracea var. capitata 'Astrus'), beterraba (Beta vulgaris 'Tall Top Early Wonder') e vagem-anã (Phaseolus vulgaris 'Alessa'). Em termos gerais, a solarização reduziu em 59% a reinfestação pela tiririca. Nas parcelas solarizadas, houve significativo incremento das produtividades de cenoura (28%), vagem (32%), beterraba (37%) e repolho (34%). A fixação biológica do nitrogênio, a julgar pela nodulação da vagem, não foi afetada pela solarização, mas a população de nematóides do solo sofreu considerável redução.

Yellow and purple nutsedges survey in the southeastern Buenos Aires Province, Argentina

A survey of 79 fields was conducted between December 1993 and January 1994, to determine the distribution and relative importance of species of the genus Cyperus, to justify developing management strategies in the southeastern of Buenos Aires Province, Argentina. Yellow and purple nutsedge were found in 43% and 9% respectively of the surveyed fields. Thirty eight per cent of the surveyed area showed a heavy infestation of yellow nutsedge, and in 90% of cases yellow nutsedge was invading fields cultivated with summer crops and associated with one or more of other seven perennial weeds, mainly bermudagrass.

Phytochemical investigation of alpine plants : the cottonsedges Eriophorum scheuchzeri Hoppe, E. angustifolium Honck. and E. latifolium Hoppe (CYPERACEAE)

Résumé: Alpine plants living at high altitudes undergo a series of climatic stress factors (chilling, enhanced UV radiation, short growing season, low nutriment supply...) which may influence their secondary compounds composition. Many publications showed in these last years that plants under stress conditions do synthesize a range of specific defence compounds (terpenes, flavonoids, coumarines...). A careful phytochemical investigation of those plants could therefore lead to the discovery of active molecules. Thus, for the biological and chemical screening, about 30 alpine plants have been collected above 2000 metres, in the alpine grass-lands. Eriophorum scheuchzeri Hoppe (Cyperaceae), not yet investigated phytochemically, revealed in its lipophilic and polar extracts the presence of various radical scavengers in a TLC autography with the DPPH (2,2-dipheny1-1- picrylhydrazyl) radical as spray reagent, as well as several antifungal compounds acitve against Cladosporium cucumerinum and Candida albi cans. The first part of this study consisted in the detection, isolation and characterization of the bioactive natural compounds present in the lipophilic extract of Eriophorum scheuchzeri. Among the eight isolated compounds, six were isoflavones. No isoflavones have been reported in the Cyperaceae family yet, nor in related families such as Poaceae or Juncaceae. Besides, isoflavones are generally rare in the plant kingdom and and they occur only in some families, such as Fabaceae, Rosaceae or Myristicaceae. In addition, out of these six isoflavones, three were new isoflavones. The known compounds were parvisoflavone A and B and cajanin which are already known isoflavones in the Fabaceae family. Two of the new isoflavones were particular, as they were C-methylated on the B-ring at the C-3' position. Methylated flavonoids are particularly rare in the plant kingdom. At present, no C-methylated isoflavones with methyl groups on the B-ring have ever been reported. The fourth new compound was a prenylated flavanone. Flavanones are also rare in the Cyperaceae family since they were found only in two genera (Cyperus and Schoenus). Finally, the widespread flavone tricin, characteristic of the Cyperaceae and Poaceae family has also been isolated. The second part of this study consisted in the characterization of the polar components present in the Me0H extract. In order to obtain mass and UV information about the secondary compounds present in the Eriophorum scheuchzeri methanolic extract, a LC-UV/DAD-APCl/MSn analysis has been performed as a first dereplication step. The UV/DAD spectra showed the presence of polyphenol compounds (phenylpropanoids and flavonoids). The LC-APCI/MSn analysis allowed the determination of the molecular weight of these compounds. Moreover, the fragmentation pattern of the [M+H]+ ions indicated presence of mono-, di- and tri-glycosides. LC-UV in combination with UV shift reagents added post-column was used in a second phase for the structural elucidation of the flavonoids. It allowed the positioning of the sugars on the aglycones. Finally, LC-NMR was used for a more detailed structural investigation of the compounds present in the crude MEOH extract. Thus, 10 fiavonoids have been totally or partially characterized by LC-UV-MS and LC-1H-RMN and UV-shift reagents added post column. However, the information obtained on-line was not always sufficient to allow a complete identification of all the compounds. Some of these compounds especially those with more than two sugar units attached to them, have been isolated in order to proceed to their complete characterization. Moreover, the Eriophorum scheuchzeri species was compared to two other species from the same genus. A LC-UV-ESI/MS analysis enabled a survey of the chemical composition of the DCM extracts of two related species E. angustifolium (Honck) and E. latifolium (Hoppe). The chromatograms of the three species showed some similarities in their flavonoid contents, especially by the recurrent presence of three compounds. The MEOH extracts of all three species have been compared by means of LC-UV-APCl/MS analyses. The chromatographic profile of all the three species showed even closer similarities than those found in the DCM extracts. E. angustifolium Honck. and E. latifolium species showed 7 compounds in common. Finally, the pure compounds obtained from the DCM (CH2Cl2) fraction were tested at different concentration, in order to evaluate their chemical and biological activities. All eight compounds showed an anti-scavenger activity against the DPPH radical, and four compounds showed antifungal activities against Cladosporium cucumerinum and Candida albicans. The pure compounds isolated from the MeOH extract were tested only for their biological activities as their antioxidant activity is already well documented in the literature. No compound showed a biological activity against Cladosporium cucumerinum and Candida albicans. Résumé: De nombreux travaux ont démontré ces dernières décennies que les plantes soumises à différents types de stress (basse température, UV, stress hydrique) synthétisent des composés secondaires (fiavonoides, coumarines, terpènes...) de protection et de défense. Les plantes d'altitude par exemple qui sont exposées à des conditions climatiques et environnementales difficiles, ont tendance à synthétiser des substances antioxydantes et antiradicalaires. Une investigation phytochirnique de ces plantes a conduit à la découverte de nouvelles molécules actives. Ainsi plusieurs plantes alpines ont été sélectionnées en fonction de leur habitat en vue de les soumettre aux tests biologiques (antifongiques) et chimiques (antiradicalaires) menés en routine dans notre laboratoire. Dans ce criblage biologique préliminaire, les extraits d'Eriophorum scheuchzeri Hoppe (Cyperaceae) ont réagi positivement aux différents tests. Il a donc été décidé d'entreprendre l'isolement des composés actifs. La première partie de ce travail a consisté à détecter, isoler et caractériser les composés naturels actifs présents dans l'extrait apolaire d' Eriophorum scheuchzeri. Parmi les huit composés isolés, quatre d'entre eux sont nouveaux. Un de ces produits est une flavanone et trois sont de nouvelles isoflavones, particulièrement intéressantes car elles possèdent des groupements C-méthylés au niveau du cycle B. Les flavonoides C-méthylés sont peu répandus dans le règne végétal et les rares exemples connus sont généralement C-méthylés sur le cycle A. Les quatre autres composés isolés n'ont jamais été décrits dans cette famille. Il s'agit d' isoflavones, les parvisoflavones A et B et la cajanine. Enfin, la flavone tricine, flavonoide caractéristique des Cyperaceae et des Poaceae a également été isolée. La deuxième partie de ce travail a consisté à caractériser les constituants polaires présents dans l'extrait methanolique. L'extrait a été analysé par chromatographie analytique couplée à différentes méthodes spectroscopiques (LC-UV-MS et LC-UV-1H RMN). De cette façon, douze flavonoides et un dérivé du phénylpropane, l'acide chlorogénique ont été identifiés. Les flavonoides tri-glycosylés ont dû être isolés afin de déterminer la nature et l'enchaînement des sucres. Finalement, l'espèce Eriophorum scheuchzeri a été comparée à deux autres espèces d' Eriophorum, soit E. angustifolium et E. latifolium. En conclusion, cette étude phytochimique a abouti à l'isolement de plusieurs nouvelles isoflavones aux activités antioxydantes et antifongiques ainsi qu'oestrogéniques.