Book of Abstracts - Geyseco

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P - Posters thesis is that ROF2 is a positive regulator of either auxin biosynthesis or perception. P07-051: CRIO4, A PATL LIKE GENE FROM SUGAR BEET IS ABLE TO CONFER TOLERANCE TO COLD STRESS BY OVEREXPRESSION Izquierdo-García A. C.* - Serrano R. - Mulet J. M. IBMCP *Corresponding author e-mail: anizgar@ibmcp.upv.es Abiotic stress is one the main limiting factors for agricultural yield. There are not extensive descriptions on which molecular processes are compromised by suboptimaltemperatures. Aiming at identifying plant genes related to cold tolerance weconstructed a cDNA library of sugar beet (Beta vulgaris) in a yeastexpression vector, isolating CRIO4 by its ability to formcolonies at 10ºC. This gene is conserved in plants and presentsseveral functional domains. Among them, a SEC14 domain. This domain issimilar to the SEC14 gene of yeast (Saccharomyces cerevisae). SEC14 encodesa Phosphatidylinositol/Phosphatidylcholine transfer protein involved incoordinate regulation of PtdIns and PtdCho metabolism. Our results indicatethat enzymatic activity should be conserved, given that our sequence datashows a conservation of the key aminoacids identified in other organisms.Aiming at confirm this observation we have purified the CRIO4 protein andperformed in vitro interaction assays with phospholipids. In addition ourstudy has identified a GOLD domain (related to protein-protein interactionin the Golgi), a Poliproline domain and several PXXP domains. CRIO4 ishomologue to the PATE- LLIN1-6 gene family of Arabidopsis thaliana. We haveidentified the members of this family which exhibit higher conservation toCRIO4, isolated homozygotic mutants, and we crossed them to obtain doublemutants. Phenotypic analysis of those double mutants, as well aslocalization data will be presented. P07-052: THE ROLE OF ABA IN THE INDUCTION OF WGA ACCUMULATION BY 24-EPIBRASSINOLIDE AND 6-BENZILAMINOPURINE IN WHEAT SEEDLING ROOTS UNDER SALINITY Bezrukova, M.* - Àvalbaev, A. - Allagulova, Ch. - Murzabaev, A, - Shakirova, F. Institute of Biochemistry and Genetics *Corresponding author e-mail: lectin@anrb.ru It is well known that gene expression and quantity of wheat germ agglutinin (WGA) are regulated by ABA which plays a key role in the regulation of plant protection to environmental stresses. Meanwhile, the essential WGA accumulation, preceded by the rapid transit increase in endogenous ABA level, is observed in wheat plants in response to biotic and abiotic stresses. Since WGA is Rab (responsive to ABA) protein it is possible to conclude that it is involved in ABA-controlled nonspecific antistress reactions of wheat plants. However we have shown that other phytohormones 24-epibrassinolide (EB) and cytokinin 6-benzylaminopurine (BAP), which possess antistress activity, induced gene expression and accumulation of WGA in wheat seedling roots. With the use of fluridone, an effective inhibitor of ABA biosynthesis, we have investigated the role of endogenous ABA in the regulation of quantitative level of WGA by EB and BAP in wheat seedlings in the normal and salt stress conditions. It was revealed that BAP-induced reversible accumulation of endogenous ABA is necessary link in the regulation by BAP of WGA level because fluridone prevented the BAP-induced WGA accumulation both in normal and stress conditions. At the same time fluridone did not affect the EB-stimulating effect on lectin accumulation in normal conditions and maintenance of WGA increased level in plants under salinity. These results serve as the proof of existence of ABA alternative pathways of hormonal regulation of concentration of WGA as component of nonspecific wheat resistance under both normal and adverse conditions and distinctions in the mechanisms of EB and BAP protective action on plants. This work was supported by Grant RFFI 08-04-01563 and Grants MK-4081.2008.4 and NSh-915.2008.4. P07-053: RNA&DNA CONTENT AFTER TEMPERATURE STRESSES IN PLANTS WITH DIFFERENT ECOLOGI- CAL STRATEGIES Negretsky, V. - Kosakivska, I. - Demirevska, K. Institute of Botany NAS Ukraine We investigated the nature of nucleic acid content changes in response to heat (2h. +40oC) and cold (2h. +2oC) temperature stresses in six plants following different ecological strategies. RNA and DNA were extracted from frozen leaf material using TRIZOL LS (Sigma, USA). The concentration of nucleic acids was measured using a Nanodrop spectrophotometer and integrity – by agarose gel electrophoresis (DNA) and capillary electrophoresis (RNA). RNA quality, integrity and concentration were determined using electrophoresis on an Agilent 2100 bioanalyzer with Nanochips. DNA quantification was performed on a Nanodrop spectrophotometer, and with 1% agarose electrophoresis with the SYBR. The results revealed both specific and non-specific properties of DNA and RNA content changes following temperature stresses. The competitor Festuca pratensis L. possessed the most stable measured levels of nucleic acids. DNA and RNA reaction was sufficiently pronounced in the ruderal Brassica napus L., which grows in low-competitiveness environments and is sensitive to biotic and abiotic stresses. Quantitative changes in ruderals were markedly stress-induced. RNA and DNA content following a temperature stresses were differentiated between C3 and C4 plants. We discuss the role of nucleic acids in response to abiotic stresses and how DNA and RNA content in plants with different strategies are affected by exposure to temperature stresses. P07-054: NTE2F OVEREXPRESSION PREVENTS G2/M CHECKPOINT ARREST NECESSARY FOR GENO- TOXIC-INDUCED PCD IN TOBACCO CELL LINE BY-2 Smetana, O. 1 - Houlné, G. 2 - Opatrny, Z. 1 - Chaboute, M. E. 2 1 Charles University 2 (IBMP Strasbourg) Various endogenous and exogenous stresses such as genotoxics induce DNA damage leading to differential gene expression, cell cycle arrest, and DNA repair or programmed cell death (PCD). The main actor responsible for sensing DNA damage induced by double strand breaks inducers (γ- and x-rays, bleomycin, zeocin) is the kinase ATM. Recently, it was reported that ATM is necessary for autophagic-type PCD induction in stem cells of Arabidopsis root meristem upon x-rays irradiation and zeocin treatment (Fulcher and Sablowski, 2009). E2F transcriptional factors play an important role during cell cycle regulation, DNA repair response or cell differentiation. In mammals and flies, E2F1 overexpression leads to increased genomic instability and subsequent cell death via transcriptional activation of several PCD genes in p53-dependent or p53-independent manner. We have investigated the effect of bleomycin (BLM) on the cell cycle progression, viability, autophagy rate, and expression of PCD-related genes in tobacco cell line BY-2. Caffeine, a specific inhibitor af ATM, is able to inhibit BLMinduced cell death. Next, to reveal the role of E2F transcriptional factor in BLM-response we generated BY-2 cell line overexpressing the only one known tobacco NtE2F transcriptional activator. Upon genotoxics, NtE2F increases genomic instability and inhibits G2/M checkpoint activation leading to decreased cell death. Thus, we suggest an important role of G2/M checkpoint during BLM-induced PCD.This work was supported by grants MSM 21-2061157 and GAUK43-259157. P
FESPB 2010 - XVII Congress of the Federation of European Societies of Plant Biology P07-055: ANALYSIS OF ABIOTIC STRESS RESPONSES USING PROTEOMIC APPROACHES Hedtmann, C. 1 - Kaspar, S. 1 - Lippmann, R. 1 - Doell, S. 1 - Matros, A. 1 - Mock HP 1 - Witzel K 2 1 Leibniz Institute of Plant Genetics and Crop Plant Research 2 Instituts für Pflanzenernährung und Bodenkunde der Universität Kiel The presentation will focus on the application of proteomics performed on model and crop plants to elucidate mechanisms and traits related to abiotic stress tolerance. Although proteomics still is limited in comprehensiveness when compared with transcriptomics, it can provide valuable information not obtained by other “omics”-techniques. We apply 2-D gel based as well as LC-MSbased approaches to study the responses of plants towards abiotic stresses on the protein level. A major focus of recent work relates to the function of co-chaperones which are induced by various stresses. These co-chaperones with homology to the human HOP protein are represented as dimers and within larger protein complexes and are located both in the cytosol and the nucleus. Central for our further work is the use of the wide genetic diversity represented e.g. in the plant genebank of our institute. As an example, barley was evaluated for contrasting salt tolerance using mapping populations. P07-056: ARSENATE SIGNALLING IN ARABIDOPSIS THALIANA Leyva Tejada, A. - Castrillo Molina, G. - Pablo Catarecha, Z. - Eduardo Sánchez, B. - Leo Del Puerto, Y. Centro Nacional de Biotecnología-CSIC *Corresponding author e-mail: aleyva@cnb.csic.es In soils arsenic is primarily in the form of arsenate [As(V)] (Tamaki and Frankenberger, 1992, Brown et al 1999). Due to its chemical similarity to phosphate (Pi), As(V) is highly toxic to plants since it is readily incorporated through the high affinity phosphate transporter (McNair et al., 1992). This transport system is induced by Pi starvation and repressed in its presence. We have demonstrated that the genes induced by Pi are properly down regulated by As (V) (Catarecha et al 2007). In fact, some of these genes (Pi transporter PHT1;1 included), are repressed by As (V) more efficiently than by Pi. Analyzing the transcriptomic profile of the As(V) response in Arabidopsis thaliana, we have found that this behavior occur at least in 10 % of the Pi-inducible-genes. These data indicates that in the case of As(V) and Pi their signaling mechanisms shares common elements. In order to identify these and others different elements, we start working in the characterization of mutants alters in the PHT1;1 repression by As(V). In this regard, we have analyzed the kinetics of PHT1;1 repression using a transgenic line expressing the gen of luciferase drive by PHT1;1 promoter. This tool provide us the possibility to concluded that this promoter are rapidly repressed by As(V) and in the case of Pi this repression is retarded (30 min and 36 h respectively). In addition, the screening of an EMS mutagenized population expressing PHT1;1:LUC led us to the identification of mutants alters in the repression by As (V). The mapping of these genes will allow us the characterization of key elements in the As (V) perception. P07-057: MITOCHONDRIAL PROTEASE, ATFTSH4, IS REQUIRED FOR ARABIDOPSIS GROWTH AND DEVE- LOPMENT UNDER CONTINUAL MODERATE HEAT STRESS. Smakowska, E.* - Kicia, M. - Gibala, M, - Janska, H. University of Wroclaw *Corresponding author e-mail: elwira@ibmb.uni.wroc.pl We found, that a loss of mitochondrial AtFtsH4 protease significantly affect Arabidopsis growth and development under continual moderate heat stress (long days at 30oC). The ftsh4 mutant plants have shorter roots and stems, beside an emergence of true leaves but not cotyledons is delayed compared with wild-type plants. The true leaves are smaller and an important characteristic of the adult leaves is the asymmetric shape and irregular serration of leaf blades. Moreover, the inflorescence development of the ftsh4 plants growing at 30oC is arrested and, as a consequence, ftsh4 plants are unable to produce seeds. Although the growth and development of the ftsh4 mutants is highly susceptible to continual moderate heat stress, the mutants are not defective in acquired thermotolerance based on physiological experiments. In agreement with this result, the transcript level of AtHsp 101, which is a cytosolic heat shock protein required for acclimation to high temperature, is similar in the wild-type and ftsh4 when plants are exposed to both a short severe heat stress (1h, 38oC) or continual moderate heat stress (1-4 weeks, 30oC). We also established that the mitochondrial AtFtsH4 protease is not a typical heat shock protein like Hsp101, because we haven’t found an increased accumulation of the AtFtsH4 transcripts upon a short time of high temperature stress. These results suggest that the thermosensitivity observed in the ftsh4 mutants is not caused by a defect in induction of cytosolic Hsp101 expression. P07-058: POLY ADP-RIBOSE POLYMERASE INVOLVE- MENT IN PROGRAMMED CELL DEATH OF TOBACCO BY-2 CELLS Locato, V. 1 - Cimini, S. 2 - Novo Uzal, E. 2 - de Pinto, M.C. 3 - Foyer, C.H. 4 - De Gara, L. 1 1 CIR-University Campus Bio-Medico di Roma 2 Departamento de Biología Animal, Vegetal y Ecología, Universidad de A Coruña 3 Dipartimento di Biologia e Patologia vegetale, Università di Bari 4 Centre of Plant Science, Leeds University, UK The term “programmed cell death” (PCD) refers to an apoptoticlike cell death that occurs during plant development and/or as consequence of various injuries. Plant PCD has many common features with animal apoptosis, but it also has some unique features related to the morphological and functional differences between the plant and animal systems. Like apoptosis, plant PCD initiation generally requires an accumulation of reactive oxygen species (ROS). Enhanced ROS production or depletion of antioxidants leads to oxidative stress, which can cause damage to cell structure and DNA. Moreover, ROS can act as molecular signals that regulate the progression of PCD. Animal and plant cells have defence and repair systems that counteract cell injury and DNA breakage. Poly-ADP ribosylation is a major process involved in DNA repair during oxidative stress and apoptosis that has been intensively studied in animal systems. However, little information is available on this process in plant cells. Here we report a study in which tobacco BY-2 cells were subjected oxidative stress-triggered PCD. Our results showed that poly ADP-ribose polymerase (PARP) activity was increased rapidly as consequence of oxidative stress. However, within hours of PCD activation PARP activity was decreased. PARP uses NAD+ to drive poly- ADP ribosylation. Thus, we suggest that regulated activation and deactivation of PARP might avoid energy depletion during PCD in order to sustain the energy-demanding death process. P07-059: LOOKING FOR CADMIUM TOLERANT MU- TANTS Agorio, A. 1 - Dardennes, J.S. 2 - Thomine, S. 1 1 Institut des Sciences du Végétal (CNRS) 2 Agroparistech Exposure to cadmium (Cd) has adverse health effects. Human uptake of Cd occurs mainly through the food-chain and tobacco smoke, as a consequence of heavy metal accumulation by crop
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FESPB 2010 - XVII Congress of the F
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PLENAR SESSION LECTUR
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PARALL SESSION LECTUR
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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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