Book of Abstracts - Geyseco

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P - Posters lignin down-regulated plants likely result from a combination of hormonal imbalance and constitutive activation of defense responses. P05-057: ANALYSIS OF CIS-ACTING ELEMENT RE- GION FOR FRUIT SPECIFICITY IN SLHD-2 PROMO- TER Kim, A.Y. – Park, S.H. – Kim, J.H. – Jeong, B.R. – Joung, Y.H.* Chonnam National University *Corresponding author e-mail: yhjoung@jnu.ac.kr Result of micro-array analysis, expression level of the histidine decarboxylase gene -2(SlHD-2) was high in tomato fruit and continuous increased during ripening. The SlHD-2 promoter was isolated from tomato (Solanum lycopersicum cv. Micro-Tom) by genome walking method. The isolated SlHD-2 promoter region was characterized here for their levels of expression and tissuespecific location of expression when transformed into tomato. GUS expression with the SlHD-2 promoter was 10 folds higher than 35S promoter in mature green and red stage fruit. On the contrary, GUS expression with the SlHD-2 was 14.4 folds lower than 35S in leaf. Histochemical staining showed that GUS was highly expressed in jelly and pericarp tissues in tomato fruit. These results show that SlHD-2 promoter was fruit specific, especially in mature stage. For analysis cis-acting elements which are related to fruit specific expression, serial deletion from 5’ region of promoter was performed. The GUS activity for the deleted promoter (▲ SlHD610) was not detected in matured fruit. In this result, cis-acting elements which are related to fruit specificity was contained in -910~-610 region. KEY WORDS: fruit specific promoter, transgenic tomato, histidine decarboxylase gene (SlHD-2) P05-058: PRODUCTION OF HEMAGGLUTININ AS VAC- CINE AGAINST INFLUENZA VIRUS USING TRANSGE- NIC PLANT Jeong, B.R. - Kim, A.Y. - Park, S.H. - Kim, J.H. – Joung, Y.H.* Chonnam National University *Corresponding author e-mail: yhjoung@jnu.ac.kr Hemagglutinin(HA) is surface protein of influenza virus. For production of influenza virus vaccine, full and partial genes of HA(A/Puerto Rico/8/34(H1N1) strain) were cloned into plant expression vector, pCAMBIA2300-HE. These full and partial genes were controlled by 35S promoter and tagged by Histidine. The recombinant vectors were transformed into tobacco (Nicotiana tabacum cv. Xanthi NC) using Agrobacterium-mediated leaf disc transformation. The transgenic tobaccolines were verified by genomic DNA PCR. HA gene expressed transgenic lines were selected by real-time PCR. HA protein was purified by TALON affinity column from these transgenic tobacco and expression level of HA gene was evaluated by ELISA. This results suggest that plant can be used as an influenza virus vaccine production system. Key worlds : Hemagglutinin, Agrobacterium mediated transformation, vaccine P05-059: CHARACTERIZATION OF TOMATO HR7 GENE(SLHR7) PROMOTER IN ARABIDOPSIS Kim, J.H. - Kim, A.Y. - Park, S.H. - Jeong, B.R. – Joung, Y.H.* Chonnam National University *corresponding author e-mail: yhjoung@jnu.ac.kr According to Micro-array data, the HR7 gene was specific expression in seed of hot pepper. For developing tomato seed specific promoter, pututive promoter region of tomato HR7 gene(SLHR7) was isolated by genome waking method. 981bp promoter include 5’-UTR was isolated and cloned into pCAM- BIA1391Z which was promoter analysis binary vector contained GUS gene. HR7 promoter was characterized here for the levels of expression and tissue specific location of expression when transformed the pCAMBIA1391z-HR7 into abidopsis. Levels of GUS expression were higher in flower then other tissues with the SLHR7 promoter. Histochemical staining showed that GUS was highly expressed at stigma and anther in flower but GUS was not detected at petal. KEY WORLDS: flower specific promoter, transgenic arabidopsis, GUS , 35S promoter P05-060: COLORECTAL CARCINOMA (CRC)-ASSOCIA- TED ANTIGEN GA733-2 GENE EXPRESSION IN TO- BACCO Park, S.H. - Kim, A.Y. - Kim, J.H. - Jeong, B.R. – Joung, Y.H.* Chonnam National University *Corresponding author e-mail: yhjoung@jnu.ac.kr Production of vaccine in plants has become an important issue in plant biotechnology field. Human colorectal carcinoma antigen GA733-2 gene and Fc region fused recombinant GA733-Fc gene were expressed into tobacco (Nicotiana tabacum cv. Xanthi-nc) plant using by Agrobacterium-mediated transformation. Plant expression vector used to this experimentation was pCAM- BIA2300-HE. The GA733-2/GA733-Fc genes were controlled by CaMV 35S promoter and tagged by histidine. Several GA733- 2/GA733-Fc transgenic tobaccos were selected. The GA733-2/ GA733-Fc proteins were detected in these transgenic plants and protein expression was quantified by ELISA. Also GA733-2/ GA733-Fc genes were expressed in tobacco using by Agroinfiltration method. Compare the expression level of GA733-2, transient expression method was more efficient then transformation. Key worlds: Human colorectal carcinoma antigen GA733-2, Agrobacterium mediated transformation, Agroinfiltration, immuno-protein, transgenic tobacco P05-061: TOOLS FOR REVERSE GENETICS AND SNP DISCOVERY Schmidt , J. – Malvoisin, P. AELRED AELRED is a recently established biotechnology start-up company, and has a technology transfer agreement with INRA and Genoplante-Valor on the latest developments of the reverse genetics TILLING® technology. AELRED provides mutagenesis and TILLING or Eco-TILLING services to its customers, either for commercial seed companies or for academic laboratories wishing to obtain new alleles of a given gene, or to confirm the function of unknown genes.On a longer term, AELRED plans to become an integrated operator in the green chemistry sector, using its technology to develop ingredients extracted from improved plants for industrial, food, pharmaceutical or cosmetic uses. AELRED is currently developing several TILLING platforms on various plant species for private and public customers, and is implementing an Arabidopsis thaliana (cv. Col 0) TILLING platform based on a collaboration with INRA-Versailles; this platform will be publicly available in 2010. AELRED hosts as well INRA’s Brassica napus (cv. Tanto) TILLING platform which is publicly open for screening. Besides, AELRED is partner in several submitted scientific projects. AELRED establishes itself as a putative partner in any project in the field of crop improvement or plant genomics, by providing the appropriate mutated plant collections and reverse genetics TILLING or Eco-TILLING screening. TILLING® r:egistered trade mark of Anawah Inc (Arcadia Biosciences). P05-062: EXPLORING ENDOGENOUS CEREAL PHYTA- SES FOR THE IMPROVEMENT OF PHOSPHORUS AND MINERAL BIOAVAILABILITY Krogh Madsen, K.* - Dionisio, G. - Bæksted Holme, I. - Bach Holm, P. - Brinch-Pedersen, H. P
FESPB 2010 - XVII Congress of the Federation of European Societies of Plant Biology University of Aarhus, Faculty of Agricultural Sciences, Dept. of Genetics and Biotechnology, Research Centre Flakkebjerg, Denmark *Corresponding author e-mail: ClausKrogh.Madsen@agrsci.dk Increasing the endogenous phytase activity in cereal grains is of importance in order to improve the sustainability of animal intensive agriculture. Furthermore it may help counter some forms of human malnutrition such as zinc deficiency which affects millions. Understanding the molecular basis for the observed variation between species and cultivars would be valuable for the effort to increase endogenous phytase activity by breeding or cisgenic approaches. Two families of phytases are known in cereals, the Multiple Inositol Phosphatases (MINPP’s) and the Purple Acid Phosphatase Phytases (PAPhy’s). Both families are highly conserved at the mature protein level and variation in specific activity within each family appears to be small. Hence we propose that the variation in phytase activity between species and cultivars are mainly attributable to gene regulation. To investigate this hypothesis we have used genomic library screening and PCR to isolate promoters of phytase genes from wheat, barley and rye. Promoters from cultivars representing the whole spectrum of grain phytase activity in wheat have been PCR amplified, cloned and sequenced. The promoters are analyzed for known regulatory elements and compared. Already, we have shown that PAPhy genes are regulated by two distinctly different types of promoters that are active primarily during grain filling and germination, respectively. In order to account for the variations in grain phytase activity we wish to correlate promoter polymorphisms with variations in expression level or pattern. These parameters are examined by proteomics, qRT-PCR, reporter gene constructs and immunolocalisation. Selected proteins are expressed in Pichia pastoris and characterized in vitro. P05-063: BIOLOGICAL EFFECT EVALUATION OF A TRITERPENIC COMPLEX TOWARDS CELL LINE DU 145 Olariu, L.* – Constantinovici, M. – Dumitriu, B. - Zglimbea, L. – Dociu, N. BIOTEHNOS S.A. *Corresponding author e-mail: lolariu@biotehnos.com Objective: Determine the triterpenic acids complex actions on DU 145, human prostate carcinoma cell line, for the purpose of studying the biological effect of this new biological active product, obtained from Salviae sp., on this neoplastic cell type. Methods: There were applied several types of cell biology specific protocols for a complete screening of its biological effects: viability and citotoxicity evaluation, using ELISA technique; proliferation, apoptosis and cell cycle analysis, using flow cytometry. The viability and cytotoxic evaluation were made after 24 h of DU 145 cells exposure to increasing doses of this complex and its carrier DMSO, ranging from 8 to 16 μM; the positive control was methothrexate 10 μM. The DNA synthesis analysis using flow cytometry were done after 48 h exposure to specific doses of triterpenic complex, DMSO and methothrexate. The statistical analysis of raw data was made with FCS Express software. Results: The triterpenic complex effect on proliferation and viability status of DU 145 cells was negative, inhibiting them in a dose dependent manner; citotoxicity was moderate until 12 μM, after that raising drastically its value. There was a significant reduction of the proliferation index which had half the value measured for untreated cells. Also this complex induced early apoptosis (30% of treated cells compared to 9% for control) and had a blocking effect on DNA synthesis (compared to DMSO and methothrexate 10 μM), both being more evident at 10 μM concentration. Conclusions: These results could be used as a starting point for more profound evaluation of biological implications of triterpenic derivates as therapeutical agents of neoplastic diseases. P05-064: SCREEN FOR DEVELOPMENTAL PHENOTY- PES IN TAXON RESTRICTED GENES Chikkaputtaiah, C.* – Donoghue, M. – Li, W. - Swamidatta, S. – Spillane, C. Genetics and Biotechnology lab, Botany and Plant science, National University of Ireland Galway, Ireland *Corresponding author e-mail: chinnatal@gmail.com Taxon restricted genes are protein coding genes which have no recognizable homologs in other species or within existing protein sequence databases. Even though they constitute up to one third of the genes found in any particular species, very little is known about their function and evolutionary genesis. Our group has defined a set of taxon restricted genes in Arabidopsis thaliana and whole genome selection scans indicated that some of the taxon restricted genes are fast evolving with a dN/dS>1 suggesting that these genes could be under positive Darwinian selection. We are performing systematic genetic, molecular and biochemical characterization of taxon restricted genes to elucidate their role in different developmental processes including reproduction and seed development in plants. P05-065: BIOCHEMICAL ANALYSIS OF DOMINANT POLLEN TYPES REPRESENTED BY LOCAL HONEY SAMPLES Tidke, J.* - Nagarkar, S.S. Department of Botany,Sant Gadge Baba Amravati University, Amravati (MS), India *Corresponding author e-mail: jaikirantidke@rediffmail.com Qualitative and quantitative biochemical analysis of pollen grains of fifteen plant species i.e. Brassica campestris, Gossypium hirsutum, Azadirachta indica, Moringa oleifera, Butea monosperma, Cajanus cajan, Syzygium cumini, Coriandrum sativum, Helianthus annuus, Parthenium hysterophorus, Tridax procumbens, Vernonia cineria, Ipomoea fistulosa, Vitex negundo and Ricinus communis was undertaken during the year 2008-2009. The pollen grains of these plant species were found to be dominantly present in the local honey samples. The pollen samples were biochemically investigated for carbohydrates and sugars, free amino acids, protein, free lipids, moisture and ash contents. The maximum amount of reducing sugar and total carbohydrates was found to be 3.08% and 5.76% in Vernonia cineria. Maximum crude protein and soluble protein i.e. 48.4% and 32.1% was found to be present in pollen grains of Parthenium hysterophorus. Free amino acids and lipid contents were encountered maximum in Helianthus annuus pollen; i.e. 3.09% and 4.10%. The maximum amount of moisture and ash was 12.83% and 7.05% in Tridax procumbens and Brassica campestris respectively. The role of some biochemical was found to be responsible for the visits of flower visiting bees in general and honey bee (A. dorsata) in particular. The data obtained through pollen biochemical analysis is being interpreted with pollen frequency class and the honey bee visits. Some pollen types such as Moringa oleifera, Helianthus annuus, Ricinus communis and Parthenium hysterophorus having more amounts of protein, carbohydrates and amino acids were dominantly represented in studied honey samples. Key Words: Biochemical, pollen, visitor relevance
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FESPB 2010 - XVII Congress of the F
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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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Book of Abstracts - Geyseco

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