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2011 PBRG Report ANNUAL REPORT 2011 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 Plant Biotechnology Research Group 8. Identification of the catabolic products of β- PBRG myrcene used as sole carbon source by Pseu- domonas sp. M1 and their correlation with bacterial gene expression. Major Achievements Objectives Pursuing our studies in the Plectranthus ge- The Plant Biotechnology Research Group nus, it was demonstrated that P. barbatus (PBRG) is a research unit on plant sciences herbal tea and its main constituent also involved in advanced education on bio- (rosmarinic acid), administered intra- technology. The main research goals, on a plu- gastrically or intraperitoneally to rats, can riannual basis, are the in vivo and in vitro sec- cross the intestinal barrier, as well as the ondary metabolites production, aiming the as- Plant Biotechnology Research Group blood brain barrier and be detected in the sessment of the National endemic aromatic/ brain, where it inhibits the enzyme acetyl- medicinal flora potentialities, the molecular cholinesterase. It was also showed that the characterization of aromatic species, focusing metabolization and bioavailability of the tea on the terpene synthase genes, the study of 7. a) Assessment of essential oils, and their components is different from the administra- plant secretory structures and the prospecting Bursaphlenchus xylophilus (Pine Wilt Nema- fractions, for their potential nematicidal proper- tion of pure compounds. In addition, protein of bioactive components. Briefly, the following tode) and Solanum tuberosum hairy roots/ ties, namely in the control of the pine wood intrinsic fluorescence analysis proved that P. topics have been under study during 2011: Meloidogyne chitwoodi (Potato-Cyst Nema- barbatus extract components and its metab- nematode, Bursaphlenchus xylophilus, b) todes) for characterization of the nematodes olites (rosmarinic acid, luteolin and apig- Research Topics Chemical characterization of Portuguese Pi- invasion mechanisms, and b) establishment of enin), found in the rat plasma, are able to nus pinaster and P. pinea volatiles from both 1. a) Chemosystematic survey of Portuguese micropropagated Thymus caespititius to assist bind to the human transport proteins albumin healthy and in lab mechanically wounded un- propolis volatiles (col. CIMO-IPB/FCUP), on the molecular elucidation of essential oil pol- and lysozyme, allowing them to be carried in b) Volatiles characterization and acaricide de- ymorphism in this species, inoculated and inoculated plants with nema- the bloodstream to different organs. P. bar- tection in Propolis from beehives maintained at todes, and c) evaluation of the histological and batus tea constituents also inhibit lysozyme 3. a) Searching for TPS genes in Thymus Algarve, c) Characterization of the volatiles cellular changes induced in pine tissues (P. activity. The inhibition of lysozyme along caespititius based on conserved regions availa- from leiva, a warm organic matterrich soil pinaster and P. pinea) by a virulent strain of B. with the radical scavenger capability, previ- ble for other Lamiaceae, starting with primer used for growing pineapples (Ananas comosus) xylophilus (collab. NemaLab – ICAM, Univ. ously shown, suggests that P. barbatus wa- design and RACE-PCR, leading to cloning the in São Miguel (Azores, Portugal) (Col. CBA), ter extract is a very promising anti- full-sequence gene, and b) construction of a Évora), and d) Characterization of the chemical compo- inflammatory raw material (col. CQB, FCUL). cDNA library of T. caespititius in order to estab- sition and biological activity of the volatiles from lish its relationship with the different chemo- Ormenis mixta and Paronychia argentea types in this species, and b) Study of heterolo- (Transnational Cooperation Portugal-Morocco), gous expression of previously identified TPS 2. a) Establishment of in vitro co-culture model genes: TPS1 and TPS2, systems of micropropagated Pinus spp./ 4. Study of the morphology, anatomy, histo- chemistry and ultrastructure of the secretory structures of medicinal and/or aromatic plants, 5. Evaluation of the biological activities of aque- ous plant extracts, infusions and decoctions, and isolated phenolic constituents, that could explain some of the traditional uses of the me- dicinal species (col. CQB-FCUL, CBA-FCUL), 6. Nutritional quality evaluation of fresh and fresh-cut fruit and their changes throughout the storage period, The essential oils from several Portuguese Miguel, M.L. Faleiro, M.J. Sousa, L.G. Ped- Thymus species, showed a quite broad vari- ro, J.G. Barroso, A.C. Figueiredo. Antimicro- PBRG ation in terms anti-acetylcholinesterase ac- bial activity, cytotoxicity and intracellular tivity. Moreover, T. zygis subsp. zygis essen- growth inhibition of Portuguese Thymus es- tial oil showed the strongest acetylcholines- sential oils. Brazilian Journal of Pharmacog- terase inhibition capacity with an IC50 = nosy, 21: 1012-1024. 1.1μg/ml. Falé P.L.V., P.J.A. Madeira, M.H. Florêncio, L. Ascensão, M.L.M. Serralheiro. Function of T. caespititius essential oil from Planalto Plectranthus barbatus herbal tea as neu- Central (Pico, Azores) and its main compo- ronal acetylcholinesterase inhibitor. Food nent, carvacrol significantly (p<0.05) inhibit- Function, 2: 130-136. ed the intracellular growth of H. pylori, and Mendes M.D., A.S. Lima, H. Trindade, A.I.D. showed no citotoxicity to the gastric cell line Correia, J.G. Barroso, L.G. Pedro, A.C. used. Biological Sciences Research Group Figueiredo. ISSR molecular characterization Plant Biotechnology Research Group The evaluation of antioxidant activity of hy- and leaf volatiles analysis of Pittosporum dro-alcoholic extracts of propolis, from differ- treatment was better for preserving firmness, undulatum Vent. naturalized in the Azores ent zones of Algarve, showed a coincidence and 2% ascorbic acid dip was better for re- archipelago (Portugal). Industrial Crops and with the highest amounts of phenols present taining the nutritional properties of fresh-cut Products, 33: 710-719. in the extracts. kiwifruit. Miguel G., C. Cruz, M.L. Faleiro, M.T.F. A high genetic variability was observed Identification and characterization of 2 puta- Simões, A.C. Figueiredo, J.G. Barroso, L.G. among individuals of Pittosporum undulatum tive TPS genes in T. caespititius, Tctps1, Pedro. Salvia officinalis L. essential oils: ef- Different solvent-extracts from several aro- that can explain the invasive ability of this Tctps2, putatively encoding for sabinene fect of hydrodistillation time on the chemical species in the Azores archipelago. The de- matic species were investigated for their an- synthase and γ-terpinene synthase, respec- composition, antioxidant and antimicrobial tailed characterization of this alien weed tioxidant and anti-inflammatory activities. S. tively. Gene structure, intron and exon activities. Natural Product Research, 25: 526 species may assist finding its potential com- officinalis extracts were the best in scaveng- placement was confirmed by comparing –541. mercial uses as well as management strate- ing DPPH radicals and superoxide anion cDNA and gDNA sequence information. The gies to help its eradication and/or control. radicals and it was verified that the time of partial characterization of Tctps3, a putative hydrodistillation influenced the antioxidant In the assays for testing the nematicidal gene coding for 1,8-cineole synthase, was activity. O. vulgare and T. mastichina ex- properties of essential oils, it was noticed also achieved. tracts were also effective as DPPH and su- that acetone seems to be an appropriate Study of Tctps1 and Tctps2 expression on T. peroxide scavengers, respectively. Deodor- alternative solvent vehicle for this kind of caespititius micropropagated cultures be- ized chloroformic extracts from T. mastichi- experiments. Moreover, the essential oil of longing to different chemotypes. na, T. camphoratus, T. carnosus, O. vulgare, Ruta graveolens showed a notorious activity Tctps1 and Tctps2 were also identified and F. vulgare and S. officinalis exhibited high (col. NemaLab – ICAM, Univ. Évora). characterized on T. caespititius, of different lipoxygenase activity inhibition. The evaluation of quality changes through- chemotypes, established in the field. out the storage period showed that citric acid Preliminary study of heterologous expres- sion of Tctps2 was performed (col. CEB/IBB, U. Minho). Selected Publications Dandlen S.A., M.G. Miguel, J.M. Duarte, M.L. Faleiro, M.J. Sousa, A.S. Lima, A.C. Figueiredo, L.G. Pedro, J.G. Barroso. Ace- tylcholinesterase inhibition activity of Portu- guese Thymus species essential oils. Jour- nal of Essential Oil Bearing Plants, 14: 140- 150. Dandlen S.A., A.S. Lima, M.D. Mendes, M.G. Publications commercial essential oils and their major com- Articles in International Peer-Reviewed Journals pounds. Molecules, 16: 7672-7690. Smail A., B. Lyoussi, M.G. Miguel, Antioxidant Ahmadi N., Negrão, S., Katsantonis, D., Frouin activity, cytotoxicity and intracellular growth inhi- activity of some Morrocan hydrosols. Journal of J., Ploux, J., Letourmy, P., Droc, G., Babo, P., bition of Portuguese Thymus essential oils. Bra- Medicinal Plants Research, 5: 6688-6696. Trindade, H., Bruschi, G.,·Greco, R., Oliveira, zilian Journal of Pharmacognosy,
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