Book of Abstracts - Geyseco

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P - Posters dent protochlorophyllide oxidoreductase (DPOR). This enzyme complex consists of three protein subunits ChlL, ChlN and ChlB, encoded by three plastid genes chlL, chlN and chlB. Using semiquantitative RT-PCR, we observed low expression of chlLNB genes in dark-grown calli. It seems, that chlLNB expression and thus Chl accumulation could be modulated by light in P. abies and L. decidua calli cultures. This hypothesis is supported by the fact, that we observed lower levels of GluTR and FLP, which probably affected Chl biosynthetic pathway at the step of ALA formation. ChlB subunit was not detected in dark-grown P. abies calli cultures. Our results and the fact, that cells of dark-grown calli contain only trace amounts of photosynthetic pigments, indicate limited ability to synthesize Chl during cultivation in the dark. P10-009: LOOKING FOR ARABIDOPSIS THALIANA HOMOLOGUES TO THE ZINNIA ELEGANS BASIC PE- ROXIDASE Cuello, J. 1 * - Herrero, J. 2 - Gómez-ros, L. 1 - Esteban Carrasco, A. 2 - Zapata, J.M. 2 - Ros Barceló, A. 1 1 Universidad De Murcia 2 Department of Plant Biology, University of Alcalá de Henares *Corresponding author, e-mail: jcuello@um.es We have previously studied the effect of auxins and cytokinins on the basic peroxidase isoenzyme from Zinnia elegans (ZePrx), an enzyme involved in lignin biosynthesis. The results showed that auxins and cytokinins induce ZePrx, similarly to the way in which they induce xylem differentiation. This hormonal response was supported by the analysis of the ZePrx promoter, which contains cis-elements directly responsive to these hormones and cis-elements targets of the plethora of transcription factors, such as NAC, MYB, AP2, MADS and class III HD Zip, which are upregulated during the auxin- and cytokinin-induced xylem differentiation. Looking for Arabidopsis thaliana homologues to the ZePrx we have found that a high degree of homology at 1D, 2D and 3D between certain peroxidases from A. thaliana and ZePrx is not always accompanied by the presence of the same regulatory cis-elements in the respective promoters. We describe the attempts made to establish the minimal structural and regulatory elements contained in the promoter region that a peroxidase involved in lignification must fulfil. This work was supported by a grant from the MEC (BFU2009- 08151)-FEDER and Fundación Séneca (08610/PI/08). P10-010: THE ACTION OF MIR169/NFY REGULATORY NETWORK IN ARABIDOPSIS THALIANA ROOT DEVE- LOPMENT Sorin, C. 1 * - Declerck, M. 2 - Lelandais-Briere, C. 1 - Christ, A. 2 - Hudik, E. 2 - Todesco, M. 3 - Weigel, D. 3 - Crespi, M. 2 - Hartmann, C. 1 1 Université Paris Diderot-Paris France /Isv Cnrs Gif Sur Yvette France 2 Isv, Cnrs Gif Sur Yvette, France 3 Max Planck Institute For Developmental Biology Tübingen, Germany *Corresponding author, e-mail: Celine.Sorin@Isv.Cnrs-Gif.Fr Roots are essential for water and nutrients acquisition in plants and root architecture is modulated by endogenous and environmental factors to optimize plant growth. microRNAs are major post-transcriptional regulators of various developmental pathways and stress responses and we have previously shown that miR169 regulation of NF-YA factors affected the formation of symbiotic nodules in legumes. To elucidate its role in root developmental plasticity in Arabidopsis, we have characterized miR169 overexpressing plants and lines with decreased miR169 activity (using a miR169 mimicry approach or mim lines, Nat. Genet. 39, 1033-7). Phenotypic analyses of mim lines suggest a role of at least one form of miR169 in root development. Expression patterns of some members of the NF-YA family of transcription factors are modulated in mim169 lines and miR169 overexpressing lines suggesting that slicing function of miR169 is partly or fully involved in the regulation mechanism. We are currently investigating NF-YA control and their role in root development. Key words: microRNA, root development P10-011: VALIDATION OF REFERENCE GENES FOR QUANTITATIVE REAL-TIME PCR DURING LEAF AND FLOWER DEVELOPMENT IN PETUNIA HYBRIDA Mallona, I. 1 * - Lischewski, S. 2 - Weiss, J. 1 - Hause, B. 2 - Egea- Cortines, M. 1 1 Universidad Politécnica de Cartagena 2 Leibniz-Institut für Pflanzenbiochemie *Corresponding author, e-mail: izaskun.mallona@upct.es Background Identification of genes with invariant levels of gene expression is a prerequisite for validating transcriptomic changes accompanying development. Ideally expression of these genes should be independent of the morphogenetic process or environmental condition tested as well as the methods used for RNA purification and analysis. Results In an effort to identify endogenous genes meeting these criteria nine reference genes (RG) were tested in two Petunia lines (Mitchell and V30). Growth conditions differed in Mitchell and V30, and different methods were used for RNA isolation and analysis. Four different software tools were employed to analyze the data. We merged the four outputs by means of a non-weighted unsupervised rank aggregation method. The genes identified as optimal for transcriptomic analysis of Mitchell and V30 were EF1α in Mitchell and CYP in V30, whereas the least suitable gene was GAPDH in both lines. Conclusions The least adequate gene turned out to be GAPDH indicating that it should be rejected as reference gene in Petunia. The absence of correspondence of the best-suited genes suggests that assessing reference gene stability is needed when performing normalization of data from transcriptomic analysis of flower and leaf development. P10-012: TERMINATION-DEPENDENT TRANSLATION OF CHLOROPLAST NDHK MRMA Sugiura M, Y.* Nagoya City University, Graduate School of natural Sciences *Corresponding author, e-mail: sugiura@nsc.nagoya-cu.ac.jp The chloroplast DNA of flowering plants is tightly packed and contains around 80 protein-coding genes. In tobacco chloroplasts, 79 protein-coding genes have so far been identified. Among them, eight genes are partially overlapped. The ndhC and ndhK genes are such examples. These genes are cotranscribed. The initiation AUG codon of ndhK mRNAs is located 4 nt upstream from the ndhC stop codon. Translational control is the major step of chloroplast gene expression. Little is known how the second cistron of overlapping gene transcripts is translated. To study mechanisms of translation unique to chloroplasts, we have developed a highly active in vitro system from tobacco chloroplasts. Using our in vitro system, mutation of the ndhC stop codon arrested translation of the ndhK cistron. The result indicated that ndhK translation depends on termination of the preceding cistron. Surprisingly, removal of the ndhC 5’-UTR and its AUG still supported substantial translation of the ndhK cistron. This translation was abolished again by removing the ndhC stop codon. Although translation of the downstream cistron of an overlapping mRNA is generally very low, we found that the ndhC/K mRNA produces NdhK and NdhC in similar amounts. Therefore, the ndhC/K mRNA is translated not only by translational coupling but also by a novel termination-dependent pathway. For the second pathway, free ribosomes are loaded on the middle of the ndhC-coding region, migrate to the ndhC stop codon and start to translate the ndhK cistron. P
FESPB 2010 - XVII Congress of the Federation of European Societies of Plant Biology P10-013: NO PROMOTES THE ACTIVATION OF A GUANYLATE CYCLASE IN NYCTINASTIC CLOSURE OF ALBIZIA LOPHANTHA LEAFLETS Bergareche, C.* - Angelo, A.P - Chellik, S. - Moysset, L. - Simón, E. Universitat de Barcelona, Facultat de Biologia *Corresponding author, e-mail: mbergareche@ub.edu NO forms complexes with plant metal containing proteins. In animals NO can initiate its biological effects through the activation of soluble guanylate cyclase (sGC); the interaction of NO with the heme ferrous iron of sGC triggers a conformational change that increases the catalysis of the second messenger cyclic GMP (cGMP) resulting in cell-specific downstream responses. Biochemical and pharmacological approaches had shown the ability of NO to induce cGMP synthesis in plant tissues. NO is involved in phytochrome mediated nyctinastic closure of Albizia lophantha leaflets. In our experimental system A.lophanta plants were maintained under 16 h light / 8 h dark cycles prior to experimental use. Pairs of leaflets were excised at 5 h of photoperiod and floated for 1 h in 10 mL control or test solutions, then irradiated with a 15 min pulse of red light (R) or a 5 min pulse of far red light (FR) and finally kept in darkness for 3 h. Exogenous application of NO donors to pairs of leaflets inhibits nyctinastic closure and NO effect is more apparent after R light irradiation. Pharmacological approaches supplying an inhibitor of nitric oxide synthase (150 mM L-NAME), results in the enhancement of nyctinastic closure, which suggests that a NOS-like enzyme may be involved in endogenous NO production response. Leaflet nyctinastic closure was not inhibited when an inhibitor of guanylate cyclase (50 mM Ly85583) was supplied, which indicates that NO increases cGMP levels. These results are corroborated by the supply of sildenafil, an inhibitor of phosphodiesterase type5 (PDE5) that produces an accumulation of cGMP and the subsequent inhibition of the nyctinastic closure. P10-014: CYTOPLASMIC/NUCLEAR PROTEINS WITH CONSERVED EUONYMUS LECTIN-LIKE DOMAINS: EVIDENCE FOR A UNIVERSAL ROLE IN EMBRYO- PHYTA. Fouquaert, E.* - Plas, K. - Van Damme E. J. M. University of Ghent *Corresponding author, e-mail: Elke.Fouquaert@UGent.be In recent years evidence has accumulated that plants synthesize well-defined carbohydrate-binding proteins (lectins) upon exposure to stresses like drought, high salt, wounding, treatment with some plant hormones or pathogen attack (1). From recent research it could be concluded that proteins with an Euonymus europaeus lectin (EUL) domain represent a new family of inducible lectins (2). Searches in the publicly available databases revealed that proteins with (an) EUL domain(s) are expressed in all Embryophyta. The family of EUL proteins is rather heterogeneous, in that some proteins consist of one EUL domain, while others comprise two in tandem arrayed EUL domains (3). Originally, these EUL proteins have been identified in rice, where they are expressed in the roots after treatment with abscisic acid and after exposure to salt-stress. An in silico expression analysis for the EUL from Arabidopsis demonstrated that this putative lectin gene is upregulated by salt-stress and osmotic stress and upon treatment with abscisic acid, suggesting that this protein plays a role in the adaptive response of plants to adverse environmental conditions. Confocal microscopy of tobacco cells, expressing GFP-fusion constructs with some EUL proteins, confirmed the nucleocytoplasmic localization of EUL proteins from rice, Arabidopsis and Euonymus europaeus. The nuclear localization together with their inducible expression indicates that these proteins with an EUL domain play an important role in regulatory processes and/or cell signalling. 1) Van Damme EJM et al., 2004, Trends in Plant Science, 9: 484-489. 2) Fouquaert E et al., 2008. Plant Physiol., 147: 1316-1324. 3) Fouquaert E et al., 2009. BMC Plant Biol., 9: 136. P10-015: AN UPSTREAM MINISATELLITE CAUSES RED APPLE FLESH COLOUR Espley, R.* - Brendolise, C. - Chagne, D. - Volz, R. - Putterill, J. - Schouten, H. - Hellens, R. - Allan, A. The New Zealand Institute of Plant & Food Research *Corresponding author, e-mail: richard.espley@plantandfood.co.nz Mutations in the genes of the anthocyanin pathway or its regulators in plants have been linked to colour phenotypes. Generally, this is a loss of function with a reduction of anthocyanin or a change in patterning. Here we describe an insertion in the upstream regulatory region of the apple anthocyanin-regulating transcription factor MYB10. This modification results in a gain of function, producing an increase in anthocyanins throughout the plant and a striking phenotype that includes red foliage and red fruit flesh. The mutation comprises a 23 base pair sequence duplicated in five tandem repeats to form a minisatellite repeat unit. We show the association between the MYB10 minisatellite duplication and the red foliage and red fruit flesh phenotype found in all apple varieties tested. Our results show that the repeat-containing promoter can act in a way that is sufficient to account for the increased MYB10 transcript levels and subsequent ectopic accumulation of anthocyanin. P10-016: STOMATOGEN, A PEPTIDE HORMONE POSI- TIVELY REGULATING STOMATA DENSITY Sakagami, Y. 1 * - Irie, K. 2 - Kakimoto, T. 3 - Kondo, T. 1 - Takada, S. 3 1 Nagoya Univ. 2 Kyoto Univ. 3 Osaka Univ. *Corresponding author, e-mail: ysaka@agr.nagoya-u.ac.jp Stomata are composed of a pair of guard cells and a pore between them, and their density and positions are regulated by developmental and environmental signals. When we overexpressed many genes coding for putative secretory proteins one-by-one in Arabidopsis, we identified a gene named STOMAGEN, which increases stomatal density when overexpressed. The STOMA- GEN gene encodes a small peptide with a putative secretory-signal sequence at its N-terminus and is expressed preferentially in mesophyll cells. This peptide belongs to the EPIDERMAL PAT- TERNING FACTOR (EPF) family of the cysteine-rich-peptides superfamily. The mature form was a C-terminal 45-amino-acid peptide (stomagen) with three intra-molecular disulfide bonds. We chemically synthesized the stomagen using solid-phase Fmoc-based chemistry, and refolded under redox-equilibrated conditions. We confirmed that the structure of the natural stomagen was identical to that of the synthetic stomagen, and determined disulfide bond positions. Stomagen treatment at very low concentrations, as low as 10 nM, increased stomatal density of wild-type Arabidopsis plants. We propose that stomagen is a mesophyll-to-epidermis signaling molecule that positively regulates stomatal density. We also suggest that stomagen increases stomatal density by competing with negative regulators EPF1 and EPF2 for the receptor-like protein TOO MANY MOUTHS. P10-017: ANALYSIS OF DROUGHT STRESS IN BARLEY BY EXPRESSION ANALYSIS AT GRAIN FILLING STA- GE Schweizer, G. 1 * - Diethelm, M. 1 - Hofmann, K. 1 - Albert, A. 2 - Winkler, J.B. 2 - Schmidhalter, U. 3 - Diego, R.P. 4 - Kleeßen, S. 4 - Lohse, M. 4 - Kersten, B. 4 - Herz, M. 2 1 LfL-Institut for Crop Science and Plant Breeding 2 Helmhotz Center Munich, Institute of Biochemical Plant Pathology (BIOP) 3 TUMunich, Department of Plant Science, Chair of Nutrition 4 MPI for Molecular Plant Physiology , Department for Bioinformatics *Corresponding author, e-mail: guenther.schweizer@LfL.bayern.de
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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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