Book of Abstracts - Geyseco

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P - Posters ²Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Germany *Corresponding author e-mail: nickivd@web.de Isoprenoids are the largest group of natural products with more than 30,000 compounds identified. The prenyl diphosphate precursors (IPP and DMAPP) for all plant isoprenoids derived from two separate, compartmentalized, biosynthetic pathways, the cytoplasmic mevalonic acid (MVA) and the plastidial methylerythritol phosphate (MEP) pathway. Beyond their specific functions in plant primary and secondary metabolism, many isoprenoids have been shown to have industrial and medical importance. In the present work, we present the generation of a platform for high level production of isoprenoids in N. tabacum by engineering the MVA and the MEP pathway to increase the IPP pool both in the cytosol and the plastids. A higher production of IPP in the cytosol could be reached by the expression of the catalytic domain of an HMG-CoA reductase (HMGR) – a key enzyme of the MVA pathway. The coexpression of a farnesyl diphosphate synthase (FPS) which uses IPP as substrate for the synthesis of isoprenoid precursors let to a 50fold increase in squalene production. A further increase in isoprenoid production rate was achieved by the upregulation of the plastidial isoprenoid pathway. The expression of a synthetic operon which was transformed to the plastom via particle bombardment, carrying ten different genes amongst others encoding enzymes involved in the MEP pathway resulted in remarkable changes in isoprenoid levels. P05-033: PROTECTION OF RARE WILD ORCHID SPE- CIES IN LATVIA EX SITU Belogrudova, I.* - Jakobsone, G. - Roze, D. - Megre, D. University of Daugavpils *Corresponding author e-mail: visio@versija.lv Ex situ conservation of terrestrial orchid species of Latvia in National Botanic Garden (NBG) is attempted in two ways – in field conditions in expositions and in the Department of Plant Diversity In Vitro - Conservation. There are three objectives for this research: elaboration of methodology for various wild terrestrial orchid species to ensure their prolonged cultivation in vitro to approach the natural development (i); optimization of acclimatization process ex vitro (ii); invention of models to create artificial plant communities in NBG, ranging into account ecology of each orchid species and similar to those in natural habitats (iii). Elaboration of methodology for in vitro cultivation involves several problems: supplementary addition of Ca 2+ in culture medium without inorganic stuffs for calciferous species, suppression of browning of culture medium resulting in plant necroses; initiation of t of new shoots to keep the microculture in rejuvenescence under study by us. The cold storage (5ºC in the dark for 5 months) was necessary for subsequent acclimatization ex vitro. The colonization with symbionts was achieved with addition to substrates the soil from natural meadow when transplanting ex vitro. Plant communities and ecology peculiarities of various orchids were documented in situ to make a cultivation strategy in expositions of NBG. Our future aim will be to establish plant communities in the field in artificial way for each in vitro acquired orchid taxon. P05-034: IDENTIFICATION AND CHARACTERIZATION OF PROTEINS INVOLVED IN RUBBER BIOSYNTHESIS IN TARAXACUM KOKSAGHYZ Hillebrand, A.¹* - Schmidt, T.¹ - Schulze Gronover, C.² - Prüfer, D.¹ ¹Department for Plant Biochemistry and Biotechnology, Westphalian, Wilhelm´s University Muenster, Germany ²Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Germany *Corresponding author e-mail: a_hill05@uni-muenster.de Natural rubber (cis-1,4 polyisoprene) is one of the most important raw materials in the world and nowadays the para rubber tree Hevea brasiliensis is the sole crop whose latex is used for commercial rubber production. As the global demand for natural rubber increases it probably can no longer be satisfied by Hevea rubber. Since the special characteristics of natural rubber, like enormous elasticity and capacitance, can not be mimicked by artificially produced polymers, there is a need for alternative rubber crops. Amongst others T. koksaghyz has been considered as a potential alternative rubber source due to its ability to produce large amounts of high quality rubber. The elucidation of rubber biosynthesis and proteins involved in this process in T. koksaghyz is crucial to improve rubber harvest and production. In H. brasiliensis a cis-prenyltransferase is supposed to be responsible for polymerization of IPP subunits to form long chains of rubber, while small rubber particle protein and rubber elongation factor influence rubber biosynthesis in a supportive manner. In T. koksaghyz we identified orthologous cDNAs encoding three cis-prenyltransferases (TkCPT1-3) and five small rubber particle proteins (TkSRPP1-5). The expression profile, localization studies as well as functional analyses implicate these proteins in rubber biosynthesis. P05-035: CROPS2INDUSTRY Rigas, S.¹* - Margaritopoulou, T.¹ – Hatzopoulos, P.¹ – Alexopoulou, E.² - Christou, M.² - Milioni, D.¹ ¹Agricultural University of Athens, Department of Agricultural Biotechnology, Athens, Greece ²Center for Renewable Energy Sources, Pikermi, Greece *Corresponding author e-mail: srigas@aua.gr Agricultural and forestry resources can provide renewable raw materials for a broad range of non-food products such as chemicals, fibres, construction materials, lubricants and fuels. The cultivation of crops for non-food use is a long-existing concept. Despite the considerable investment in research and development, little progress has been made towards the commercial exploitation of such products. Crops2Industry overall objective is to examine the potential use of non-food crops for selected industrial applications namely oils, fibres, resins, pharmaceuticals or other specialized products within the EU27 context. Biotechnology will play a key role in producing biomaterials that conform to technical specifications. This project aims to outline and prioritise crops-to-products schemes suitable for individual Member States, which will support a sustainable, economically viable and competitive European bio-based industry and agriculture. The project consortium is composed of fourteen partners from nine European countries, involving four Universities, five research organizations/institutes and five companies. The partners will gather data on the potential establishment of non-food crops in combination with biotechnology to overcome breeding constraints, the assessment of market needs, product quality standards, prices and yield costs. Emphasis will be given to the dissemination of information and data to farmers and end-users in terms of the most promising non-food crops. Crops2Industry is expected to assess whether and under which terms Europe has the potential and technical competence to develop a competitive bio-industry fed by sustainable agriculture and become a world leader in the field of bio-based products. P05-036: MICROPROPAGATION OF SCOTS PINE FROM SEEDLING EXPLANTS Lebedev, V.* - Schestibratov, K. Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry *Corresponding author e-mail: vglebedev@mail.ru Conifer clonal forestry as a form of plantation forestry has great potential advantages. However conifers are most recalcitrant objects for cultivation in vitro. Scots pine is one of the most wides- P
FESPB 2010 - XVII Congress of the Federation of European Societies of Plant Biology pread forest trees in Russia. We investigated influence of various factors, such as basal nutrient media, cytokinins, ethylene, activated charcoal, light intensity and photoperiod length on bud induction, shoot elongation and rooting of explants from young seedlings of Scots pine. Basal media used in our study include MS, PM1, SH, WPM, TE, MCM and DCR media. WPM and MS media induced vitrification of most explants. Use of TE medium resulted in explant necrosis. SH medium gave the best results. BA, kinetin, zeatin, thidiazuron and 2iP at various concentrations were tested for the induction of adventitious buds. Kinetin and 2iP had little bud-inducing effect compared to BA and zeatin. Optimal conditions for pulse treatment of explants with BA were defined. Addition of activated charcoal inhibited bud production but promoted shoot elongation. Root initiation up to 44% were obtained after exposition on NAA-containing medium. Rooted plantlets were transferred to the greenhouse and acclimatized with 90% survival. Our results may be applied for development of clonal micropropagation method of Scots pine using vegetative tissue explants from mature trees. P05-037: OPTIMIZING DNA DELIVERY ON SOMATIC EMBRYOGENIC TISSUE OF MARITIME PINE Blasco Carlos, M.* - Humanez, A. - Mendoza-Poudereux, I. - Muñoz-Bertomeu, J. - Almazan, V. - Brisa, C. - Segura, J. - Arrillaga, I. Universitat de València *Corresponding author e-mail: miblas@alumni.uv.es Maritime pine (Pinus pinaster Ait.) is a characteristic species in Mediterranean forests, with its main populations located in the Iberian Peninsula. The species is also the most advanced conifer model for genomic research in Europe. Genetic transformation is the best tool to allow gene functional analysis and for rapidly increasing yield and wood quality. The objective of this work is to develop strategies to efficiently deliver DNA into somatic embryogenic tissue by Agrobacterium tumefaciens coculture techniques. Embryogenic lines were initiated from immature megagametophytes and maintained by 2-week subcultures on Litvay medium (mLV). Embryogenic callus was resuspended in liquid mLV and mixed with the same volume of the bacterial suspension. After infection, cells were recovered on filter paper and transferred to the same medium for coculture. Transient Gus Assays were performed following standard protocols afterthree days of coculture. Factors tested were: Agrobacterium strain (AGL1, EHA105 and LBA4404); infection by sonication or vacuum infiltration; coculture time (10, 30 and 60 min); and method to recover embryogenic material after infection (by a low-pressure pulse on a Buchner funnel or poured on filter paper over a pile of paper towels). Among factors tested, AGL1 strain applied under vacuum infiltration (1 min) followed by 10 min infection and further recover of cells on a filter paper over paper towels produce the higher number of blue foci per plate (average of 130). Additiona l r esults on stable integration will be presented. This work is funded by the Spanish MICINN and FEDER funds (AGL2007-66345-CO2-02); Generalitat Valenciana (Prometeo 2009/075) and a Research Fellowship to M. B. P05-038: STRATEGY FOR PRODUCTION OF RICE WITH INCREASED ISOFLAVONE GENISTEIN LEVEL BY METABOLIC ENGINEERING OF PHENYLPROPA- NOID PATHWAY Park, H.* - Choi, M.S. – Lee, Y.Y. – Shon, J.Y. – Choi, I.S. – Yun, H.T. – Lee, J.Y. – Kim, Y.J. National Institute of Crop Science, RDA *Corresponding author e-mail: parkhm2002@korea.kr Isoflavonoids are a diverse group of plant natural products synthesized from phenylpropanoid pathway which play important roles in plant growth and development. There have been considerable attentions as health-promoting nutraceuticals because of their antioxidant and estrogenic anticancer activity. In our study, we attempted to develope genisteinenriched rice for recommendable daily consumption through engineering the isoflavone pathway. Both overexpression and RNAi suppression strategies were used to manipulate the expression of several genes encoding key enzymes in the flavonoids/ isoflavonoids pathway in transgenic rice. Rice plants were transformed with two soybean (Glycine max L) isoflavone synthase (GmIFS1, 2) genomic DNA under the control of the seed specific rice globulin promoter. HPLC-MS analysis demonstrated the presence of genistein as the major isoflavone metabolite in the transgenic plants. Substantial amounts of genistein (up to 87.0 μg/g FW) were found in seeds. However, the amounts of genistein showed annual variation in the field condition. For producing transgenic rice seeds with high level of genistein in a stable manner, we transformed maize C1 and R-S genes that together activate most structural genes in the flavonoid biosynthetic pathway. Furthermore, we constructed a RNAi vector to suppress the flavanone 3-hydroxylase (F3H) gene expression for blocking anthocyanin synthetic pathway. We obtained transgenic rice plants harboring RNAi-F3H vector or C1 and R-S overexpression vector and then we pyramided these three genes (IFS1/ / C1R-S/RNAiF3H) by crossing. These plants will be used for further analysis of isoflavone detection and molecular characterization. (Supported by RDA Biogreen 21 and NICS grant) P05-039: MOLECULAR CHARACTERIZATION AND DE- VELOPMENT OF MOLECULAR MARKER FOR BREE- DING LOW ALLERGY SOYBEAN CULTIVARS BY NU- LLIFYING MAJOR ALLERGEN P34 Choi , M.* - Jeong, K.H. – Lee, S.K. – Seo, M.J. – Yun, H.T. – Park, H.M. – Kim, Y.H. National Institute of Crop Science (RDA) *Corresponding author e-mail: mschoi73@korea.kr Soybean (Glycine max (L.) Merr.) is an important source of vegetable oil and high protein. Use of soybean meal by the food industry is increasing, but severely limiting dietary choices and the quality of life of food-allergic individuals. Gly m Bd 30K (P34) is known as the main seed allergens in soybean-sensitive patients. The objective of this work was to determine the molecular basis of the low mutation of soybean P34 and to design molecular marker for the selection of the causative mutations for wild homozygous, heterozygous and mutant homozygous. Using soybean genome assembly, we knew that soybean P34 genes are existing 2 copies in LG A1 and 1 copy in LG A2 in soybean genome. We confirmed three copies of P34 genes in soybean genome by Southern blot analysis and found that all of those genes are expressing during the seed filling stage through RT-PCR analysis. Especially, Glyma08g12270 of those was expressed at significantly higher level compared with Glyma08g12280 and Glyma05g29130. However this gene was not expressed in the low-P34 germplasm accessions. We developed a co-dominant marker based on the sequence of Glyma08g12270 containing a four-base pair insertion at the P34 start codon. Also, we made a polyclonal antibody for investigation of P34 protein levels. Using a co-dominant marker and a polyclonal antibody, polymorphism and amount of protein for Glyma08g12270 were analyzed in F2 and F3 generation crossing PI 567476 and Hwanggumkong, Korean cultivar. As results, the polymorphism analysis was accustomed to a difference of protein level of wild homozygous, heterozygous and mutant homozygous. P05-040: INCREASED VITAMIN E CONTENT IN SOYBEAN PLANTS OVEREXPRESSING HGGT GENE FROM RICE ENHANCED TOLERANCE TO ABIOTIC STRESS AND ANTIOXIDANT ACTIVITY Kim, Y.* - Park, H.M. - Lee, Y.Y. – Kim, Y.H. – Choi, M.S. – Jeong, K.H. – Lee, S.K. – Seo, M.J. – Yun, H.T. National Institute of Crop Science, RDA *Corresponding author e-mail: kimyuh77@korea.kr
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POST 04 POSTERS • Environmental S
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Author Index Aarts, M.G. S18-002 Ab
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Cattivelli, L. P01-031 Cawly, J. P1
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Gallego, B. P17-037 Gallego, P.P. P
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Kersten, B. P10-017 Keskin, O. P05-
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Miguel, C. S02-001, P01-122 Milhinh
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Ramiro, M. P09-018 Ramon, M P13-002
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Tanimoto, E. P01-045 Tarakanov, I.
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