PBRGPlant Biotechnology Research Group Institute for Biotechnology and Bioengineering Departamento de Biologia Vegetal Faculdade de Ciências de Lisboa Campo Grande 1749-016 Lisbon Portugal http://cbv.fc.ul.pt

ANNUAL REPORT

INSTITUTE FOR BIOTECHNOLOGY AND BIOENGINEERING

2009

PBRG | Plant Biotechnology Research Group

Agro-Food

Biotechnology

The thematic area of Agro-Food Biotechnology is focused on the application of expertises in func-tional, comparative and evolution genomics, molecular genetics, to biological systems of interest in agriculture, animal biotechnology and food processing. IBB is committed to develop R&D activities within the thematic area of Agro-Food Biotechnology on four major topics: A) Comparative and Evolution Genomics and Genetic Variability to analyze the polymorphisms of relevant genes in crops, fruit trees and forestry species and of the somatropic traits in bovidae and suinae species. Genomic approaches to understand nutrient control of gene expression, to control the food chain and to identify GMOs. B) Genetic Resources use for improvement of agriculture and forestry in Portugal, aiming at the exploitation of the natural Flora heritage with economic interest. C) Plant and Animal Biotechnology for the production of bioactive metabolites; marker assisted se-lection; plant resistance to herbicides and in vitro plant micropropagation. D) Food Bioprocess Technology for wine, olive oil, food and beverage processing; microbial food spoilage control.

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Plant Biotechnology Research Group

Objectives

The research activities of PBRG in Agro-Food Biotechnology have been focused on the in vitro cultures, with particular emphasis on hairy roots cultures as natural product producing systems, namely essential oils. In addition, chemical and molecular characterization of endemic plant spe-cies as well as the study of secretory structures, viewing a better knowledge, valuation and conser-vation of the national plant biodiversity resources, with potential economic interest is another topic on which PBRG has been particularly concerned.

Research Topics

1. Phytochemical and Molecular Analysis - Evaluation of the correlation between the volatiles composition and molecular markers and, in addi-tion, the molecular characterization of terpene synthase genes aiming at clarifying how the pro-duction of terpenes, namely monoterpenes, is regulated. Identification of interesting genotypes, particularly those that produce essential oils con-taining bioactive compounds.

2. In vitro Culture - Increase and/or modify the volatiles production by hairy root cultures and mi-cropropagation, by manipulation of the culture medium and of the photoperiod conditions, differ-ent nitrogen sources and ratios, addition of pre-cursors or elicitors. On the hairy root cultures, induction of regenerants, culturing in a bioreactor

and in a two-phase system and biotransformation will also be tested. Understand the hairy root cul-ture behaviour and reveal some advantages of this in vitro system for the production of the differ-ent types of secondary metabolites that can be found in plant volatiles.

3. Biology of Plant Secretion - Survey of the differ-ent types of secretory structures in the vegetative and reproductive organs of medicinal and aro-matic plants. Morpho-anatomical characterization of these glands, ultrastructural study of their cells and histochemical identification of the main classes of compounds present in the secreted material. The overall integration of these data en-hances our understanding of cell secretory proc-esses and of physiological and ecological funtions of plant secretions. Major Achievements

�� RAPDs and ISSRs were used as molecular markers and combined with the chemical analysis of the volatiles, aiming at characteriz-ing Thymus caespititius from the islands of Corvo, Flores and Graciosa (Azores). Thirty-one individuals were analyzed. Molecular data discriminated assessments from Corvo and Flores which clustered separately from plants collected on Graciosa. Volatile data grouped plants in a similar way, depending on their oil

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type. The first data on terpene synthases genes in Th. caespititius aiming at elucidating the molecular basis of product specificity attrib-uted to different chemotypes revealed that the genomic sequences of three partial genes from chemotypes were almost identical. Protein pre-diction revealed a partial gene structure, in-cluding four exons which, when aligned, showed a number of conserved residues, such as the DDXXD motif. Other motifs could play a role in monoterpene specificity.

�� Chemical and molecular analyses were per-formed, aiming the characterization of Junipe-rus brevifolia, an Azorean endemic species, evaluating the relationship between both data sets. Forty-two individual twigs samples were collected on seven Azorean islands (S. Miguel, Terceira, S. Jorge, Pico, Faial, Flores and Corvo). Cluster analysis of chemical data showed a high correlation among all samples. Cluster analysis of molecular data showed a

moderate correlation between accessions and plants were grouped according to their geo-graphical location, not showing identical clus-tering as with the volatile oil profiles.

�� A combined analysis of Laurus azorica volatile oils, RAPDs and ISSRs data, was performed to evaluate the relationship between both data sets. Cluster analysis of molecular data showed a moderate correlation. Plants were grouped mainly according to their geographical location, providing no straight correlation be-tween molecular and chemical clusters, sug-gesting that different molecular tools and addi-tional chemical profiling should be performed, in particular with Graciosa and S. Miguel ac-cessions.

�� Cluster analysis of the volatiles composition of forty-eight samples of seven liverwort Radulaspecies collected on Azores and Madeira ar-chipelagos and in mainland Portugal, as well as of one species from Switzerland, resulted in two major clusters. One cluster included the oil samples of Radula aquilegia from Madeira, R.jonesii, R. holtii, R. nudicaulis, R. lindenbergi-ana as well as R. complanata from Switzer-land. The second cluster included the oil sam-ples of R. aquilegia from the Azores as well as R. carringtonii and R. wichurae. The cluster analysis allowed distinction of all studied mor-phological species, indicating the chemotax-onomical value of the volatile oil characters.

�� In collaboration with NemaLAb – ICAM, Uni-versidade de Évora, it was demonstrated that the damage inflicted by Bursaphelenchus xylo-

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philus, the pine wood nematode (PWN) in the stem tissues of Pinus pinaster seems to de-pend of plant fitness and of environmental con-ditions such as temperature and water avail-ability. At an “early stage” of infection, when no external symptoms were visible, PWNs were already observed within the lumen of cortical resin ducts. At an “advanced stage” of infec-tion, when external symptoms were plainly visible a severe tissue degradation occurred. Cavities with irregular boundaries developed from degraded resin ducts, the number of nematodes increased dramatically and all pine tissues were damaged.

�� In collaboration with NemaLAb – ICAM, Uni-versidade de Évora, twenty seven essential oils, isolated from plants of eleven families of the Portuguese flora, were screened for their nematicidal activity against the pinewood nematode (PWN), Bursaphelenchus xylophi-lus. High nematicidal effect was achieved with essential oils of Chamaespartium tridentatum, Origanum vulgare, Satureja montana, Thymbra capitata and Thymus caespititius, with an esti-mated minimum inhibitory concentration rang-ing between 0.097 and 0.374 mg/ml and LC100 between 0.858 and 1.984 mg/ml. The other essential oils yielded weak or no activity. Five oils with nematicidal activity against PWN were reported for the first time.

�� In collaboration with CDCTPV and CBME, FCT, Universidade do Algarve, the essential oils of several Thymus species, collected in different regions of Portugal, including Azores islands, were evaluated for preventing lipid peroxidation and scavenging free radicals, hy-droxyl and superoxide anion. T. zygis subsp. zygis oil possessed the best capacity for pre-venting lipid peroxidation. The oils isolated

from T. zygis subsp. sylvestris were the most effective for scavenging free radicals. The best capacity for scavenging hydroxyl radicals was found in the oils of T. camphoratus and T. caespititius. For the superoxide anion scaveng-ing determination, the highest inhibition per-centages were registered for the essential oils of T. zygis subs. sylvestris and T. capitellatus.

�� In collaboration with CDCTPV and CBME, FCT, Universidade do Algarve, the antioxidant, anti-inflammatory and antiacetylcholinesterase activities of essential oils and remaining water-distillation extracts of eight aromatic plants (Dittrichia viscosa, Foeniculum vulgare, Ori-ganum vulgare, Salvia officinalis, Thymbra capitata, Thymus camphoratus, T. carnosus and T. mastichina) were studied. The DPPH radical scavenging activity evaluation revealed that the remaining water-distillation extracts were more effective than the essential oils. O.vulgare remaining water-distillation extract, the one with the highest amount of phenols, showed the best antioxidant activity, while the most effective essential oils were those of T.capitata and O. vulgare. T. capitata remaining water-distillation extract showed the best su-peroxide anion scavenging activity. F. vulgare remaining water-distillation extract and essen-tial oil showed the best capacity for inhibiting lipoxygenase activity. Th. mastichina, S. offici-nalis, T. capitata and Th. camphoratus essen-tial oils were the most effective in the assess-ment of antiacetylcholinesterase activity.

�� In collaboration with Centro de Química e Bio-quimica, Universidade de Lisboa and pursuing our studies in Plectranthus genus, it was showed that decoction of leaves from P. barba-tus had high inhibition of acetlylcholinesterase

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and antioxidant activity. The main constituents identified in this extract (rosmarinic acid, scutellarein 4’-methyl ether 7-O-glucuronide and (16S)-coleon E) also showed antiacetyl-cholinesterase activity. In addition, leaf aque-ous extracts of P. barbatus and P. ecklonii and rosmarinic acid had an inhibitory effect on the viability, glucosyltransferase activity and biofilm formation of two cariogenic bacteria (Strepto-cccus sobrinus and S. mutans). Therefore, the use of these leaf decoctions could be promis-ing for the prevention and control of several oral diseases, since they have low toxicity and demonstrated significant inhibition of water insoluble glucans synthesis and reduce biofilm production, critical factors for dental plaque formation.

�� In collaboration with Dept. de Engenharia Química e Biológica, IST, supercritical fluid extraction (SFE) of the volatile oil fromSatureja montana L. was performed under dif-ferent conditions. The results were compared with those obtained for the essential oil iso-lated by hydrodistillation (HD) and the antioxi-dant activities evaluated. The volatile oil ob-tained by SFE was the most effective, with the lowest EC50 and the highest protector factor. The volatile oil is 15 times richer in thymo-quinone than the essential oil (HD). This com-pound is of great importance due to its antioxi-dant, neuroprotective, and anti-cancer activi-ties. The combination of carvacrol+thymol+ thymoquinone in volatile oil may be responsi-ble for the increase in the antioxidant activity when compared to HD, which demonstrates that SFE improves value to the final product. The morphology of the glandular trichomes of

S. montana and the effect of the grinding proc-ess on them was also evaluated by SEM.

�� The biotransformation capacity of Levisticum officinale and Anethum graveolens hairy root cultures was studied by evaluating the effect of the addition of menthol or geraniol on morphol-ogy, growth, and volatiles production. With Levisticum officinale hairy roots, morphology and growth was not influenced by substrate addition and no new volatiles were detected after menthol addition and the addition of gera-niol induced the production of six new volatiles. Following the addition of � glycosidase to the remaining distillation water, analysis of the ex-tracted volatiles showed that lovage hairy roots were able to convert both substrates and their biotransformation products into glycosidic forms. With Anethum graveolens hairy root cultures, substrate addition had no negative influence on growth, the relative amount of menthol quickly decreased and the biotransfor-mation product menthyl acetate was formed. The added geraniol quickly decreased along-side with the production of 10 new products.

�� Hairy root cultures of Solanum tuberosum and Digitalis purpurea (in collaboration with Prof. Dr W. Kreis, Erlangen University, Germany) were established, as well as the micropropagation of Pinus pinaster, P. pinea, Eucalyptus globulus and Pyrus communis. Particularly, the S. tube-rosum hairy root cultures and the micropropa-gated P. pinaster and P. pinea plants will be used as models for studying the infection mechanism of plant nematodes in co-cultures.

Selected Publications

Bentes, J., Miguel, M. G., Monteiro, I., Costa, M., Figueiredo, A. C., Barroso, J. G., Pedro, L. G., Ital. J. Food Sci., 21, 183-195. Falé, P. L., Madeira, P., Borges, C., Ascensão, L., Araújo, M. E., Florêncio, M. H., Serralheiro, M. L. M., Food Chem., 114, 798-805. Faria, J. M. S., Nunes, I. S., Figueiredo, A. C., Pedro, L. G., Trindade, H., Barroso, J. G., Bio-technol. Lett., 31, 897-903. Nunes, I. S., S. Faria, J. M., Figueiredo, A. C., Pedro, L. G., Trindade, H., Barroso, J. G., Planta Med., 75, 387-391. Trindade H., Costa, M. M., Lima, A. S. B., Pedro, L. G., Figueiredo, A. C., Barroso, J. G., Biochem. Syst. Ecol., 37, 670-677.

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Articles in International Peer-Reviewed

Journals

Bentes, J., Miguel, M. G., Monteiro, I., Costa, M., Figueiredo, A. C., Barroso, J. G., Pedro, L. G. (2009) Antioxidant activities of the essen-tial oils and extracts of Portuguese Thymbra capitata and Thymus mastichina. Ital. J. Food Sci., 21, 183-195.

Carmo-Silva, A. E., Francisco, A., Powers, S. J., Keys, A. J., Ascensão, L., Parry, M. A., Arrabaça, M. C. (2009) Grasses of different C4 metabolic subtypes reveal traits related to the intensity of drought stress in their natural habi-tats: changes in leaf dry matter, water potential, structure and amino acid content. Am. J. Bot., 96, 1222-1235.

Falé, P. L., Madeira, P., Borges, C., Ascen-são, L., Araújo, M. E., Florêncio, M. H., Ser-ralheiro, M. L. M. (2009) Rosmarinic Acid, scutellarein 4'-methyl ether 7-O-glucuronide and (16S)-Coleon E are the main compounds re-sponsible for the antiacetylcholinesterase and antioxidant activity in herbal tea of Plectranthus barbatus (“Falso Boldo”). Food Chem., 114, 798-805.

Faria, J. M. S., Nunes, I. S., Figueiredo, A. C., Pedro, L. G., Trindade, H., Barroso, J. G. (2009) Biotransformation of menthol and gera-niol by hairy root cultures of Anethum graveolens: effect on growth and volatile com-ponents. Biotechnol. Lett., 31, 897-903.

Figueiredo, A. C., Sim Sim, M., Barroso, J. G., Pedro, L. G., Esquível, M. G., Fontinha, S., Luís, L., Martins, S., Lobo, C., Stech, M. (2009) Liverwort Radula species from Portugal: chemotaxonomical evaluation of volatiles com-position. Flavour Frag. J., 24: 316-325.

Grosso, C., Figueiredo, A. C., Burillo, J., Mainar, A. M., Urieta, J. S., Barroso, J. G., Coelho, J. A., Palavra A. M. F. (2009) Enrich-ment of the thymoquinone content in volatile oil from Satureja montana using supercritical fluid extraction. J. Sep. Sci., 32, 328-334.

Grosso, C., Oliveira, A. C., Mainar, A. M., Urieta, J. S., Barroso, J. G., Palavra, A. M. F. (2009) Antioxidant activities of the supercritical and conventional Satureja montana extracts. J. Food Sci., 74, 713 - 717.

Grosso, C., Teixeira, G., Gomes, I., Martins, E. S., Barroso, J. G., Pedro, L. G., Figueiredo, A. C. (2009) Assessment of the essential oil composition of Tornabenea annua, T. insularis and T. tenuissima fruits from Cape Verde Is-lands. Biochem. Syst. Ecol., 37, 474-478.

Matos, F., Miguel, M. G., Duarte, J., Venâncio, F., Moiteiro, C., Correia, A. I. D., Figueiredo, A. C., Barroso, J. G., Pedro, L. G. (2009) Antioxidant capacity of the essential oils from Lavandula luisieri, L. stoechas ssp. lusitanica, L. stoechas ssp. lusitanica x L. luisieri and L. viridis grown in Algarve (Portugal). J. Essent. Oil Res., 21, 327-336.

Publications

Mendes, M. D., Trindade, H., Figueiredo, A. C., Barroso, J. G., Fontinha, S. S., Pedro, L. G. (2009) Volatile and molecular characterization of two Portuguese endemic species: Angelica lignescens andMelanoselinum decipiens. Biochem. Syst. Ecol., 37, 98-105.

Mendes, M. D., Trindade, H., Figueiredo, A. C., Pedro, L. G., Barroso, J. G. (2009) Chaero-phyllum azoricum Trel. grown in the Azores ar-chipelago (Portugal): Evaluation of the genetic diversity using molecular markers and compari-son with volatile oils profiles. Flavour Frag. J., 24, 259-265.

Nunes, I. S., S. Faria, J. M., Figueiredo, A. C., Pedro, L. G., Trindade, H., Barroso, J. G. (2009) Menthol and geraniol biotransformation and glycosylation capacity of Levisticum offici-nale hairy roots. Planta Med., 75, 387-391.

Pereira, A. L., Figueiredo, A. C., Barroso, J. G., Pedro, L. G., Carrapiço, F. (2009) Volatile compounds from the symbiotic system Azolla filiculoides - Anabaena azollae – bacteria. Plant Biosyst., 143, 268-274.

Trindade H., Costa, M. M., Lima, A. S. B., Pedro, L. G., Figueiredo, A. C., Barroso, J. G. (2009) A combined approach using RAPD, ISSR

and volatile analysis for the characterization of Thymus caespititius from Flores, Corvo and Graciosa islands (Azores, Portugal). Biochem. Syst. Ecol., 37, 670-677.

M.Sc. Thesis

Lima, A. S. B., “Estratégias para caracterização molecular de espécies aromáticas da flora Portuguesa“, MSc Thesis, Faculdade de Ciências, Universidade de Lisboa, Lisbon (supervisor: Helena Trindade, FCUL, Lisbon).

Pimpão, R. “Estudo da síntese de polifenóis em Juniperus sp. endémicos de Portugal”, MSc Thesis, Faculdade de Ciências, Universidade de Lisboa, Lisbon (supervisors: A. Cristina Figueiredo, FCUL, Lisbon and Ricardo Boavida Ferreira do ITQB/UNL, Lisbon).

Santana, J. C. C., “Análise cromatográfica e identificação de biomarcadores de envelhecimento de documentos gráficos”, MSc Thesis, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Lisbon (supervisors: A. Cristina Figueiredo, FCUL, Lisbon and Elvira Gaspar, FCT/UNL, Lisbon).

International Conferences Albano, S. M., Miguel, M. G., Pedro, L. G., Barroso, J. G., Figueiredo, A. C., “Antioxidant, anti-inflammatory and antiacetylcholinesterase activities of essential oils and remaining water-distillation extracts of some aromatic plants”, 40th International Symposium on Essential Oils, Savigliano, Itália, p. 127.

Dandlen, S. A., Figueiredo, A. C., Pedro, L. G., Barroso, J. G., Miguel, M. G., Faleiro, M. L., “Antibacterial activity comparison of several Portuguese Thymus essential oils”, 57th Inter-national Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research | GA 2009, Genebra, Suíça, p. 1060.

Dandlen, S. A., Miguel, M. G., Faleiro, M. L., Pedro, L. G., Barroso, J. G., Figueiredo ,A. C., “Antioxidant activity from five Portuguese thyme species essential oils”, 40th International Sym-posium on Essential Oils, Savigliano, Itália, p. 128.

Dandlen, S. A., Miguel, M. G., Faleiro, M. L., Pedro, L. G., Barroso, J.G., Figueiredo, A. C., “Capacidade de captação de radicais NO· invitro dos óleos essenciais de algumas espécies

Portuguesas de Thymus”, 2º Congresso Iberoamericano de Fitoterapia, 1º Congresso de Fitoterapia da SPFito e 5º Congreso de la SE-FIT | Fito 2009, Lisboa, Portugal, p. 152.

Figueiredo, A. C., Garcia, C., Sim-Sim, M., Sérgio, C., Pedro, L. G., Barroso, J. G., “Volatiles from Plicanthus hirtellus and Radula boryana grown in São Tomé and Príncipe”, 40th International Symposium on Essential Oils, Savigliano, Itália, p. 125.

Figueiredo, A. C., Mendes, M. D., Pedro, L. G., Barroso, J. G., Guia, H., Miguel, M. G., Cruz, C., Antunes, M. D., Cavaco, A. M., “Propolis volatile composition from beehives maintained in Algarve (Portugal)”, 40th International Symposium on Essential Oils, Savigliano, Itália, p. 124.

Grosso, C., Teixeira, G., Gomes, I., Martins, E. S., Barroso, J. G., Pedro, L. G., Figueiredo, A. C., “Evaluation of the essential oil composi-tion of fruits of three endemic species of Torna-benea from Cape Verde Islands”, 57th Interna-tional Congress and Annual Meeting of the Soci-ety for Medicinal Plant and Natural Product Re-search | GA 2009, Genebra, Suíça, p. 928-929.

Poster Presentations

Oral Communications International Conferences

Lopes V. R., Barata, A. M., Farias, R., Men-des, M. D., Pedro, L. G., Barroso, J. G., Fi-gueiredo A. C., “Evaluation of the morphologi-cal and essential oil variability from nine Portu-guese fennel (Foeniculum vulgare Mill.) acces-sions”, 4th International Symposium Breeding Research on Medicinal and Aromatic Plants | ISBMAP2009, Ljubljana, Slovenia, p. 16. – (invited lecture)

Lima, A. S., Barbosa, P., Mota, M., Barroso, J. G., Pedro, L. G., Figueiredo, A. C., “Volatiles characterization of Pinus pinaster and Pinus pinea inoculated with the Pine Wood Nematode (Bursaphelenchus xylophilus)”, Gordon Conference on Floral and Vegetative Volatiles, Oxford, UK.

Lima, A. S., Trindade, H., Figueiredo, A. C., Barroso, J. G., Pedro, L. G., “Molecular and volatiles characterization of Portuguese Thymus caespititius chemotypes”, 4th International Sym-posium Breeding Research on Medicinal and Aromatic Plants | ISBMAP2009, Ljubljana, Slo-venia, p. 49.

Lima, A. S., Trindade, H., Figueiredo, A. C., Barroso, J. G., Pedro, L. G., “Volatile charac-terization and molecular polymorphism evalua-tion among Azorean Laurus azorica”, 57th Inter-national Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research | GA 2009, Genebra, Suíça, p. 929.

Lima, A. S., Trindade, H., Figueiredo, A. C., Barroso, J. G., Pedro, L. G., “Combined vola-tile and molecular analysis of Juniperus brevifo-lia, an endemic Azorean species”, 40th Interna-tional Symposium on Essential Oils, Savigliano, Itália, p. 126.

Lima, A. S., Trindade, H., Figueiredo, A. C., Barroso, J. G., Pedro, L.G., Marques, M. I., “Characterization of a putative monoterpene synthase genomic sequence of Portuguese Thy-mus caespititius (Lamiaceae)”, Plant Genomics European Meeting | PlantGem, Lisboa, Portugal, p. 198.

Luís L., Sim-Sim, M., Stech, M., Esquível, M. G., Fontinha, S., Lobo, C., Garcia, C., Martins, S., Vieira, C., Barroso, J., Pedro, L., Figueiredo, A. C., “Characterization of the genus Radula (Marchantiophyta) in Portugal. A molecular, phytochemical and morphological approach”, 7th Biennal Conference of the Sys-tematics Association, Leiden, Holanda, p. 86.

Marin, M., Dulectic-Lausevic, S., Ascensão, L., “The histochemical analysis of Satureja sub-spicata Bartl. ex Vis. glandular trichomes”, 5th Balkan Botanical Congress, Belgrade, Serbia.

Meira, R., Francino, D. M. T., Ascensão, L. “Ontogenia e histoquímica dos tricomas secretores de Chamaecrista dentata (Legumino-sae, Cesalpinioideae)”, 60º Congresso Nacional de Botânica, Bahia, Brasil.

International Conferences (cont) Mendes, M. D., Trindade, H., Figueiredo, A. C., Pedro, L. G., Barroso, J. G. “Trichomes morphology and essential oils composition in micropropagated vs field grown Thymus caes-pititius plants”, 4th International Symposium Breeding Research on Medicinal and Aromatic Plants | ISBMAP2009, Ljubljana, Slovenia, p. 77.

Mendes, M., Trindade, H., Figueiredo, A. C., Barroso, J. G., Pedro, L. G., “Caracterização volátil e molecular de indivíduos de Crithmum maritimum colhidos no arquipélago dos Açores (Portugal)”, 2º Congresso Iberoamericano de Fitoterapia, 1º Congresso de Fitoterapia da SPFito e 5º Congreso de la SEFIT | Fito 2009, Lisboa, Portugal, p. 124.

Miguel, M. G., Dandlen, S. A., Figueiredo, A. C., Pedro, L. G., Barroso, J. G., Marques, M. H. “Comparative evaluation of the antioxidant activities of thymol and carvacrol and the corre-sponding b cyclodextrin complexes”, III Interna-tional Symposium on Aromatic and Medicinal Plants | SIPAM 2009, Djerba, Tunisia, p. 135.

Mohammed, A. D., Miguel, M. G., Antunes, M. D., Figueiredo, A. C., Pedro, L. G., Barroso, J. G. “Antioxidant activity of Thymbra capitata es-sential oil in meat-treated oil”, III International Symposium on Aromatic and Medicinal Plants | SIPAM 2009, Djerba, Tunisia, p. 134-135.

Mota, L., Barbosa, P., Mota, M., Ascensão, L., “Histological changes induced by Bursaphelen-chus xylophilus in seedling tissues of Pinus pi-naster”, 48th Annual Meeting of the Society of Nematologists and 12th Biennial Meeting of the Soil Ecology Society, Vermont, USA.

Mota, L., Galupa, R., Barbosa, P., Mota, M., Ascensão, L., “Anatomical changes induced by Bursaphelenchus xylophilus in tissues of Pinus

pinaster”, 22nd New Phytologist Symposium – Effectors in plant-microbe interactions, Ver-sailles, France.

Moura, B., Nogueira, T., Ascensão, L., “Morfologia, anatomia e histoquímica das estruturas secretoras de três espécies de Hypericum da flora portuguesa”. 2º Congresso Iberoamericano de Fitoterapia, 1º Congresso de Fitoterapia da SPFito e 5º Congreso de la SEFIT | Fito 2009, Lisboa, Portugal, p. 128.

Porfírio, S. A. P., Falé, P. L. V., Ascensão, L., Serralheiro, M. L. M., “Digestion of Plectran-thus barbatus decoction: citotoxicity, in vitro me-tabolism and the remaining antiacetylcholi-nesterase and antioxidant activities”, Functional Molecules from Natural Sources, Oxford, UK.

Rodrigues, L., Duarte, A., Figueiredo, A. C., Teixeira, G., Moldão, M., Monteiro, A., “Antibacterial and antifungal activity of essential oils of Thymbra capitata grown in Portugal”, 2º Congresso Iberoamericano de Fitoterapia, 1º Congresso de Fitoterapia da SPFito e 5º Congreso de la SEFIT | Fito 2009, Lisboa, Portugal, p. 121.

Poster Presentations (cont)

Rodrigues, L., Póvoa, O., Teixeira, G., Fi-gueiredo, A. C., Moldão, M., Monteiro, A., “Micromorphology of trichomes and composition of the essential oil of Mentha pulegium L. from Portugal”, III International Symposium on Aro-matic and Medicinal Plants | SIPAM 2009), Djerba, Tunisia, p. 52.

Sullivan, S., Carneiro, A. P., Pedro, L. G., Barroso J. G., Figueiredo A. C., “Caract-erização química de óleos essenciais isolados de fitoterápicos e comerciais utilizados em aromaterapia: Eucalyptus globulus, Foeniculum vulgare e Mentha piperita”, 2º Congresso Iberoamericano de Fitoterapia, 1º Congresso de Fitoterapia da SPFito e 5º Congreso de la SEFIT | Fito 2009, Lisboa, Portugal, p. 124.

Vieira, P., Barbosa, P., Lima, A. S., Dias, L. S., Tinoco, M. T., Pedro, L. G., Figueiredo, A. C., Barroso, J. G., Mota, M., “Nematicidal activ-ity and composition of the essential oils from the Portuguese aromatic flora against the pinewood nematode (Bursaphelenchus xylophilus)”, Inter-national Symposium on Pine Wilt Disease, Nan-jing, China, p. 76.

Other publications

Faria, J. M. S., Sanches, J., Lima, A. S., Mendes, M. D., Figueiredo, A. C., Trindade, H., Pedro, L. G., Barroso, J. G., “Caracteri-zação dos óleos essenciais de espécies do género Eucalyptus da Mata Experimental do Escaroupim”, Actas do 6º Congresso Florestal Nacional, pp. 658-661.

Figueiredo A. C., Barroso, J. G., Pedro, L. G., Luís, L., Sim-Sim, M., “Chiloscyphus polyan-thos, a new addition to the Flores Island (Azores) bryoflora”, Portugaliae Acta Biol., 23, 241-242.

Sim-Sim M., Bergamini, L., Luís, L., Martins, S., Lobo, C., Fontinha, S., Figueiredo, A. C., Barroso, J. G., Pedro, L. G., Stech, M., “Epiphytic bryophyte communities on Madeira: Effects of tree species on bryophyte species richness”, XVII Simpósio de Botânica Criptogâmica, Tomar, p. 74-75.

Sim-Sim, M., Stech, M., Esquível, M. G., Luís, L., Fontinha, S., Lobo, C., Garcia, C., Martins, S., Vieira, C., Barroso, J., Pedro, L., Figuei-redo, A. C., “Molecular, phytochemical and morphological characterisation of the liverwort genus Radula in Portugal mainland, Azores and Madeira”, XVII Simpósio de Botânica Criptogâmica, Tomar (Oral presentation).