2009 PBRG Report - Center for Plant Biotechnology

Transcrição

2009 PBRG Report - Center for Plant Biotechnology
ANNUAL REPORT
INSTITUTE FOR BIOTECHNOLOGY AND BIOENGINEERING
PBRG | Plant Biotechnology Research Group
PBRG
Plant 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
2009
Agro-Food
Biotechnology
The thematic area of Agro-Food Biotechnology is focused on the application of expertises in functional, 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 selection; 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.
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 species as well as the study of secretory structures,
viewing a better knowledge, valuation and conservation 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 addition, the molecular characterization of terpene
synthase genes aiming at clarifying how the production of terpenes, namely monoterpenes, is
regulated. Identification of interesting genotypes,
particularly those that produce essential oils containing bioactive compounds.
2. In vitro Culture - Increase and/or modify the
volatiles production by hairy root cultures and micropropagation, by manipulation of the culture
medium and of the photoperiod conditions, different nitrogen sources and ratios, addition of precursors or elicitors. On the hairy root cultures,
induction of regenerants, culturing in a bioreactor
moderate correlation between accessions and
plants were grouped according to their geographical location, not showing identical clustering as with the volatile oil profiles.
x
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 between molecular and chemical clusters, suggesting that different molecular tools and additional chemical profiling should be performed,
in particular with Graciosa and S. Miguel accessions.
x
Cluster analysis of the volatiles composition of
forty-eight samples of seven liverwort Radula
species collected on Azores and Madeira archipelagos 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. lindenbergiana as well as R. complanata from Switzerland. The second cluster included the oil samples of R. aquilegia from the Azores as well as
R. carringtonii and R. wichurae. The cluster
analysis allowed distinction of all studied morphological species, indicating the chemotaxonomical value of the volatile oil characters.
x
In collaboration with NemaLAb – ICAM, Universidade de Évora, it was demonstrated that
the damage inflicted by Bursaphelenchus xylo-
and in a two-phase system and biotransformation
will also be tested. Understand the hairy root culture behaviour and reveal some advantages of
this in vitro system for the production of the different types of secondary metabolites that can be
found in plant volatiles.
3. Biology of Plant Secretion - Survey of the different types of secretory structures in the vegetative
and reproductive organs of medicinal and aromatic 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 enhances our understanding of cell secretory processes and of physiological and ecological funtions
of plant secretions.
Major Achievements
x
RAPDs and ISSRs were used as molecular
markers and combined with the chemical
analysis of the volatiles, aiming at characterizing Thymus caespititius from the islands of
Corvo, Flores and Graciosa (Azores). Thirtyone 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
type. The first data on terpene synthases
genes in Th. caespititius aiming at elucidating
the molecular basis of product specificity attributed to different chemotypes revealed that the
genomic sequences of three partial genes from
chemotypes were almost identical. Protein prediction revealed a partial gene structure, including 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.
x
Chemical and molecular analyses were performed, aiming the characterization of Juniperus 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
Plant Biotechnology Research Group
PBRG
Plant Biotechnology Research Group
x
philus, the pine wood nematode (PWN) in the
stem tissues of Pinus pinaster seems to depend of plant fitness and of environmental conditions such as temperature and water availability. 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 infection, 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.
x
x
In collaboration with NemaLAb – ICAM, Universidade 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 xylophilus. High nematicidal effect was achieved with
essential oils of Chamaespartium tridentatum,
Origanum vulgare, Satureja montana, Thymbra
capitata and Thymus caespititius, with an estimated minimum inhibitory concentration ranging 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, hydroxyl and superoxide anion. T. zygis subsp.
zygis oil possessed the best capacity for preventing lipid peroxidation. The oils isolated
x
In collaboration with CDCTPV and CBME,
FCT, Universidade do Algarve, the antioxidant,
anti-inflammatory and antiacetylcholinesterase
activities of essential oils and remaining waterdistillation extracts of eight aromatic plants
(Dittrichia viscosa, Foeniculum vulgare, Origanum 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 superoxide anion scavenging activity. F. vulgare
remaining water-distillation extract and essential oil showed the best capacity for inhibiting
lipoxygenase activity. Th. mastichina, S. officinalis, T. capitata and Th. camphoratus essential oils were the most effective in the assessment of antiacetylcholinesterase activity.
In collaboration with Centro de Química e Bioquimica, Universidade de Lisboa and pursuing
our studies in Plectranthus genus, it was
showed that decoction of leaves from P. barbatus had high inhibition of acetlylcholinesterase
S. montana and the effect of the grinding process on them was also evaluated by SEM.
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 antiacetylcholinesterase activity. In addition, leaf aqueous 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 (Streptocccus sobrinus and S. mutans). Therefore, the
use of these leaf decoctions could be promising 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.
x
In collaboration with Dept. de Engenharia
Química e Biológica, IST, supercritical fluid
extraction (SFE) of the volatile oil from
Satureja montana L. was performed under different conditions. The results were compared
with those obtained for the essential oil isolated by hydrodistillation (HD) and the antioxidant activities evaluated. The volatile oil obtained by SFE was the most effective, with the
lowest EC50 and the highest protector factor.
The volatile oil is 15 times richer in thymoquinone than the essential oil (HD). This compound is of great importance due to its antioxidant, neuroprotective, and anti-cancer activities. The combination of carvacrol+thymol+
thymoquinone in volatile oil may be responsible 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
x
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 morphology, 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 geraniol induced the production of six new volatiles.
Following the addition of ̻glycosidase to the
remaining distillation water, analysis of the extracted 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 biotransformation product menthyl acetate was formed.
The added geraniol quickly decreased alongside with the production of 10 new products.
x
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. tuberosum hairy root cultures and the micropropagated 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., Biotechnol. 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.
Biological Sciences Research Group
Plant Biotechnology Research Group
PBRG
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 scavenging determination, the highest inhibition percentages were registered for the essential oils
of T. zygis subs. sylvestris and T. capitellatus.
and volatile analysis for the characterization of
Thymus caespititius from Flores, Corvo and
Graciosa islands (Azores, Portugal). Biochem.
Syst. Ecol., 37, 670-677.
Publications
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 essential 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 habitats: 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., Ascensão, L., Araújo, M. E., Florêncio, M. H., Serralheiro, M. L. M. (2009) Rosmarinic Acid,
scutellarein 4'-methyl ether 7-O-glucuronide and
(16S)-Coleon E are the main compounds responsible 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 geraniol by hairy root cultures of Anethum
graveolens: effect on growth and volatile components. Biotechnol. Lett., 31, 897-903.
Figueiredo, A. C., Sim̻Sim, 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 composition. 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) Enrichment 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 Islands. 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.
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).
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
and
Melanoselinum decipiens. Biochem. Syst. Ecol.,
37, 98-105.
Mendes, M. D., Trindade, H., Figueiredo, A.
C., Pedro, L. G., Barroso, J. G. (2009) Chaerophyllum azoricum Trel. grown in the Azores archipelago (Portugal): Evaluation of the genetic
diversity using molecular markers and comparison 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 officinale 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
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).
Oral Communications
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
International Conferences
Lopes V. R., Barata, A. M., Farias, R., Mendes, M. D., Pedro, L. G., Barroso, J. G., Figueiredo A. C., “Evaluation of the morphological and essential oil variability from nine Portuguese fennel (Foeniculum vulgare Mill.) accessions”, 4th International Symposium Breeding
Research on Medicinal and Aromatic Plants |
ISBMAP2009, Ljubljana, Slovenia, p. 16. –
(invited lecture)
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 Symposium Breeding Research on Medicinal and
Aromatic Plants | ISBMAP2009, Ljubljana, Slovenia, p. 49.
Poster Presentations
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 waterdistillation 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 International 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 Symposium 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· in
vitro 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 SEFIT | 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 composition of fruits of three endemic species of Tornabenea from Cape Verde Islands”, 57th International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research | GA 2009, Genebra, Suíça, p. 928-929.
Lima, A. S., Trindade, H., Figueiredo, A. C.,
Barroso, J. G., Pedro, L. G., “Volatile characterization and molecular polymorphism evaluation among Azorean Laurus azorica”, 57th International 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 volatile and molecular analysis of Juniperus brevifolia, an endemic Azorean species”, 40th International 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 Thymus 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 Systematics Association, Leiden, Holanda, p. 86.
Marin, M., Dulectic-Lausevic, S., Ascensão,
L., “The histochemical analysis of Satureja subspicata 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 (Leguminosae, Cesalpinioideae)”, 60º Congresso Nacional
de Botânica, Bahia, Brasil.
Other publications
Poster Presentations (cont)
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 caespititius 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 corresponding b̻cyclodextrin complexes”, III International 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 essential 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 Bursaphelenchus xylophilus in seedling tissues of Pinus pinaster”, 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, Versailles, France.
Faria, J. M. S., Sanches, J., Lima, A. S.,
Mendes, M. D., Figueiredo, A. C., Trindade,
H., Pedro, L. G., Barroso, J. G., “Caracterização dos óleos essenciais de espécies do
género Eucalyptus da Mata Experimental do
Escaroupim”, Actas do 6º Congresso Florestal
Nacional, pp. 658-661.
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.
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