Book of Abstracts - Geyseco

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P - Posters This work was supported by CICYT AGL2006-01743/AGR and 11883/PI/09 (Fundación Séneca, Agencia de Ciencia y Tecnología de la Región de Murcia) P17-009: AN AM FUNGUS AND A PGPR INFLUENCE PHYSIOLOGICAL AND MOLECULAR MECHANISMS INVOLVED IN ALLEVIATION OF LETTUCE PLANTS SUBJECTED TO ELEVATED CO2 AND DROUGHT Kohler, J.* - Alguacil M.M. - Caravaca, F. - Roldán, A. CEBAS-CSIC *Corresponding author, e-mail: jkohler@cebas.csic.es Arbuscular mycorrhizal (AM) symbiosis and plant-growth-promoting rhizobacterium (PGPR) can alleviate the effects of water stress in plants but it is unknown whether these benefits can be maintained at elevated CO2. Therefore, we carried out a study where seedlings of Lactuca sativa were inoculated with the AM fungus (AMF) Glomus intraradices N.C. Schenk & G.S. Sm. or the PGPR Pseudomonas mendocina Palleroni and subjected to two levels of watering and two levels of atmospheric CO2 to ascertain their effects on plant physiological parameters and gene expression of one PIP aquaporin in roots. The inoculation with PGPR produced the greatest growth in lettuce plants under all assayed treatments as well as the highest foliar potassium concentration and leaf relative water content under elevated [CO2] and drought. However, under such conditions, the PIP2 gene expression remained almost unchanged. G. intraradices increased significantly the AMF colonization, foliar phosphorus concentration and leaf relative water content in plants grown under drought and elevated [CO2]. Under drought and elevated [CO2], the plants inoculated with G. intraradices showed enhanced expression of the PIP2 gene as compared to P. mendocina or control plants. Our results suggest that both microbial inoculation treatments could help to alleviate drought at elevated [CO2]. However, the PIP2 gene expression was increased only by the AMF but not by the PGPR under these conditions. P17-010: A NEW APPROACH FOR THE COMPREHEN- SIVE ANALYSIS OF MICROCOMMUNITIES IN PLANT ECOLOGY Moshynets, O.* 1 - Shpylova, 1 . - Kosakivska, I. 1 - Kordium, V. 1 - Bogaerts, L. 2 - Potters, G. 2 - Koza, A. 3 - Spiers, A. 3 1 Institute of Molecular Biology and Genetics NAS of Ukraine 2 Department of Bioscience Engineering, University of Antwerp, Groenenborgerlaan 171 B-2020, Antwerp, BELGIUM 3 The SIMBIOS Centre, University Abertay Dundee, Scotland, UK *Corresponding author, e-mail: moshynets@gmail.com Plants are constantly interacting with a wide range of microbes which effect plant growth, development and response. However, comparatively little is known about ecological interactions in the phytosphere mostly due to the technical failing of current methods like the lack, or loss, of spatial informaton which is essential to the understanding of the interractions operating over the micrometer range. Here we offer a novel approach to assess the microflora in specific phytoenvironments. The QS (quasi-substratum) approach uses pieces of plastic film which are placed in the environment and allowed to develop microbial communities on their surfaces. X-ray microtomography can be used to determine the position of the film in situ with respect to local structures of the micro-environments. When recovered, the films can be stained to allow direct CLSM imaging of the surface-associated microcommunity while direct extraction of the microbial DNA enables the use of molecular methods. The QS approach was tested in the rhizosphere of Brassica napus where the interactions between the GFP-tagged rhizobacterium Pseudomonas fluorescens SBW25 and the native soil microbes were studied, as well as in the study of the endophytes of bamboo plants Phyllostachys atrovaginata. In the later case, endophytic micro-communities were observed microscopically and total microbial community 16S DNA was extracted. The subcloning and subsequent identification of species will be performed at later stage. The QS approach enables the use of modern imaging technologies coupled with molecular technologies, allowing comprehensive investigation of microbial diversity in the phytosphere, which allows more in-depth analysis of the interaction between microbes and plants in the future. P17-011: REACTIVE ELECTROPHILE SPECIES OR- CHESTRATE BACTERIAL-INDUCED STOMATAL CLO- SURE IN ARABIDOPSIS THALIANA. Montillet, J.* - Leonhardt, N. - Mondy, S. - Tranchimand, S. Cea Cadarache-Ibeb-Lems *Corresponding author, e-mail: Jean-Luc.Montillet@Cea.Fr Stomatal closure has been found as a part of plant innate immune response allowing restriction of bacterial invasion at leaf surface (Melotto et al. 2006). In this context, participation of different signalling molecules such as abscisic acid (ABA), salicylic acid (SA) and nitric oxide (NO) has been suggested. Our present work describes evidence indicating that reactive electrophile species (RES) originating from lipoxygenase activity are key players involved in this physiological process. The role of RES has been assessed in Arabidopsis challenged with strains of Pseudomonas syringae or treated with flg22 a pathogen-associated molecular patterns (PAMP). Our data show that steps at the upstream of this new signalling cascade differ from those triggered by ABA. Reference: Melotto et al. 2006, Plant stomata function in innate immunity against bacterial invasion. Cell 126, 969-980. P17-012: RHIZOBIAL ALDEHYDE OXIDASE AND LE- GUME NODULATION Mercedes Lucas, M.* - Torres, MJ. - Paradela C. - Rincón, A. - Coba de la Peña, T. - Pueyo, J.J. - Delgado, MJ *Corresponding author, e-mail: mlucas@ccma.csic.es The symbiotic association between legumes and rhizobia leads to the development of a symbiotic organ, the nodule. Phytohormones such as the auxin indol acetic acid (IAA), synthesized by both symbionts, play an important role in legume nodulation. To date, little is known about the auxin biosynthesis pathways during root nodule development. In plants, the enzyme aldehyde oxidase (AO, EC 1.2.3.1) catalyzes the last step of IAA biosynthesis via the Trp-dependent indole-3-pyruvic acid pathway. We have previously described the presence of plant AOs in nodules of Medicago truncatula and Lupinus albus and proposed their involvement in the regulation of nodule development (1). In addition to plants, AOs have also been found in some microorganisms, including rhizobia (1, 2). We have performed a search in the genome sequence of Sinorhizobium meliloti 1021 by using the protein sequences corresponding to the small, medium and large subunits (AodS, AodM, AodL) of AO from Methylobacillus sp. KY4400 (2). We have identified three genes in S. meliloti 1021 clustered in the same transcriptional order found for aodS, aodM and aodL genes in Methylobacillus sp. In order to investigate the involvement of these genes in the symbiotic interaction S. meliloti-Medicago sativa, mutants in the first gene of the transcriptional unit have been constructed by marker exchange. The symbiotic phenotype of one of the strains obtained, mutated in the aodS-like gene, is being investigated. (1) Fedorova et al. 2005. MPMI 18: 405–413. (2) Yasuhara et al. 2005. Biosci. Biotechnol. Biochem. 69: 2435- 2438. P17-013: IMMEDIATE SUPPRESSION OF ROOT-BASED IMMUNITY GUARANTEES COLONISATION SUCCESS OF THE SYMBIONT PIRIFORMOSPORA INDICA Schaefer, P.* Research Centre for BioSystems, Land Use and Nutrition; Justus Liebig University *Corresponding author, e-mail: patrick.schaefer@agrar.uni-giessen.de P
FESPB 2010 - XVII Congress of the Federation of European Societies of Plant Biology Piriformospora indica is root-colonizing fungus that transfers several benefits to the colonised plants, e.g. enhanced biotic and abiotic stress resistance or tolerance, respectively. The fungus is member of the order Sebacinales, which was found to house various forms of mycorrhizal fungi. Several microorganisms are able to establish beneficial or pathogenic interactions with plants. In general, adaptation and specialisation evolved by beneficial or pathogenic microbes results in narrowed host range. This is most extremely demonstrated by the biotrophic lifestyle of certain leaf colonizing fungi that showplant species-dependent reproduction. Thorough epifluorescence and transmission electron microscopy-based studies in Arabidopsis roots defined an initial biotrophic preceding a cell death-associated colonisation phase of P. indica implicating a distinct host-microbe communication. Despite its biotrophism, P. indica colonises roots of an extraordinarily broad spectrum of monocot and dicot plants. Results presented indicate the importance of the suppression of early defense response, especially of the oxidative burst, to be significantly associated with its ability to establish a compatible root interaction. In addition, we will discuss the relevance of leaf defense determinants in root-based immunity. P17-014: COMPARATIVE STUDY OF SOIL-BASED AND COMMERCIAL FORMULATIONS OF ARBUSCULAR MYCORRHIZAL FUNGI IN LETTUCE SUBJECTED TO SALT STRESS Garmendia López, I.* Universidad de Alicante *Corresponding author, e-mail: idoia.garmendia@ua.es Oltra Cámara M A (Oltra M A) Mangas Martín V J (Mangas V J) Arbuscular mycorrhizal fungi (AMF) can contribute to the salinity resistance of host plants by improving nutritional status, particularly of P and N, enhancing osmotic adjustment, increasing warter use efficiency and resucing oxidative damage. Our objective was to compare the effect of soil-based (Glomus intraradices and G. mosseae) and commercial AMF inocula (formulations 1 and 2) in salt stress tolerance of lettuce. No effect on shoot biomass was found with mycorrhizal inoculation under non-saline conditions and root biomass was significantly reduced. Only plants inoculated with formulation 1stimulated shoot dry weight not related with greater root AMF colonization. P and K concentration in leaves were improved bymycorrhizal association. Salinity did not reduce leaf relative water content and we observed no osmotic adjusment in leaves from non-mycorrhizal plants. However, root dry biomass and its starch content decreased, while leaf starch and root soluble sugar concentrations were enhanced in non-inoculated plants under saline conditions. Lettuce inoculated with formulation 2 and soil-based G. intraradices showed the highest root colonization percentages. Neverthelessn, one of the mycorrhizal treatments induced a significant improvement on the growth of lettuce subjected to salinity. The severity of the salt stress applied and the effectiveness of mycorrhizal inocula ameliorating its effect is discussed. This work was supported by Universidad de Alicante (UAUSTI09/04). P17-015: IDENTIFICATION AND FUNCTIONAL CHA- RACTERIZATION OF PUTATIVE PHOSPHORYLATION TARGETS OF SLCIPK INVOLVED IN IMMUNE RES- PONSE IN PLANTS Gutiérrez-Beltrán, E.* - del Pozo, O. - de la Torre, F. IBVF-US/CSIC *Corresponding author, e-mail: emgube@gmail.com Two novel components of the Pto-mediated hypersensitive response (HR) signal transduction pathway, a calcium sensor (calcineurin B-like; CBL) and a CBL-interacting protein kinase (CIPK) were identified in a random gene silencing screening (VIGS) in Nicotiana benthamiana. Recently, we have also demonstrated the participation of their tomato orthologs, SlCIPK and SlCBL during bacterial speck disease symptom development caused by Pseudomonas syringea pv tomato (P. syringae pv tomato) in tomato. Considering that SlCIPK is an active kinase that plays a role in R-gene signal transduction pathways, it might elicit a phosphorylation cascade in response to pathogen attack. In order to identify putative phosphorylation targets and upstream regulation elements of SlCIPK, we conducted two parallel yeast two-hybrid screenings using SlCIPKWT and SlCIPKCA (a constitutive active mutant version) respectively as baits. A normalized prey library generated from tomato was used. From the screening we obtained several different interacting proteins. The selection of candidates was based on three different criteria: (1) implication in plant immunity through reverse genetic approaches (VIGS), (2) capability to interact with SlCIPK in vivo and (3) putative biological function. Silencing susceptible tomato plants with one of these clones dramatically reduced P. syringae pv tomato growth four days after infection. Therefore, this clone named CIP29 (CIPK Interacting Protein) was selected for further in detail characterization. We have found that CIP29 is phosphorylated in vitro by SlCIPK. This clone represents a new phosphorylation target for the CIPK family, and the first identified substrate different of a membrane channel. P17-016: TOWARDS IDENTIFICATION OF SYSTEMIC ACQUIRED RESISTANCE SIGNALLING PARTNERS DOWNSTREAM FROM ENHANCED DISEASE SUSCEP- TIBILITY1 Vlot, C. 1 * - Jordá, L. 2 - Colby, T. 2 - Wenig, M. 1 - Knappe, C. 1 - Rietz, S. 2 1 Helmholtz Zentrum Muenchen; Institute of Biochemical Plant Pathology 2 Max Planck Institute for Plant Breeding Research *Corresponding author, e-mail: corina.vlot@helmholtz-muenchen.de Wittek, F (Helmholtz Zentrum Muenchen; Institute of Biochemical Plant Pathology) Breitenbach, H (Helmholtz Zentrum Muenchen; Institute of Biochemical Plant Pathology) Mackey, D (The Ohio State University; Department of horticulture and crop science) Parker, J E (Max Planck Institute for Plant Breeding Research) Systemic acquired resistance (SAR) is a long lasting, broad spectrum disease resistance that is induced in the systemic tissue of plants undergoing a localized immune response. Often, the primary immune reaction is accompanied by programmed cell death, the hypersensitive response (HR), of cells in and around the site of infection. During the HR resistance-inducing signals are transmitted that can for instance be found in petiole exudates from Arabidopsis thaliana leaves that are infected with Pseudomonas syringae encoding the HR-inducing effector AvrRpm1. Petiole exudates from infected wild type (wt) A. thaliana trigger PR1 gene expression in healthy wt plants. By contrast, similar exudates from infected enhanced disease susceptibility1 (eds1) mutant plants do not, although the AvrRpm1-induced HR is intact in this mutant. This indicates that EDS1 is required for SAR signal generation and/or transmission. We are now using conditional expression of AvrRpm1 to induce an HR in wt and eds1 mutant plants and could show that this induces SAR-like disease resistance in systemic non-AvrRpm1- expressing tissues of wt plants. Moreover, candidate SAR signalling components were identified by 2D gel and GC-MS analysis of apoplast extracts from AvrRpm1-expressing wt plants as compared to similar extracts from eds1 mutants. We are currently analyzing pathogen resistance and in particular SAR in Arabidopsis and tobacco over expressing the corresponding putative SAR signalling genes. P17-017: MOLECULAR CLONING AND CHARACTE- RIZATION OF A PATHOGENESIS-RELATED PROTEIN CSPR10 FROM CROCUS SATIVUS
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Page 206 and 207: Author Index Aarts, M.G. S18-002 Ab
Page 208 and 209: Cattivelli, L. P01-031 Cawly, J. P1
Page 210 and 211: Gallego, B. P17-037 Gallego, P.P. P
Page 212 and 213: Kersten, B. P10-017 Keskin, O. P05-
Page 214 and 215: Miguel, C. S02-001, P01-122 Milhinh
Page 216 and 217: Ramiro, M. P09-018 Ramon, M P13-002
Page 218 and 219: Tanimoto, E. P01-045 Tarakanov, I.
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