Book of Abstracts - Geyseco

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P - Posters P13-035: ISOLATION OF ARABIDOPSIS MUTANTS IN THE EXPRESSION OF A GENE FOR OIL SYNTHESIS BY USING HIGH-THROUGHPUT REAL-TIME BIOLUM Kawai, T. 1 - Onai, K. 2 - Matsumoto, T. 1 - Hashimoto, M. 1 - Maeo, K. 1 - Ishiura, M. 2 - Nakamura, K. 1 1 Lab. Biochem. Grad. Sch. Bioagr. Sci., Nagoya University 2 Cent. Gene Res., Nagoya Univ. and SENTAN, JST During the maturation of Arabidopsis seeds, a transcription factor WRI1/ASML1 coordinately activates many genes involved in fatty acid synthesis in the plastid. However, expression of genes for triacylglycerol (TAG) assembly in the endoplasmic reticulum is regulated differently. Arabidopsis plants with a luciferase (LUC) reporter gene under the promoter of a gene for diacylglycerol acyltransferase (DGAT1::LUC) showed strong LUC expression in maturing seeds and weaker expression in young leaves, that were similar to the expression patterns of DGAT1 mRNA. Mutants with altered LUC expression were screened with high-throughput real-time bioluminescence monitoring system, which can automatically monitor bioluminescence of 1,920 or 19,200 seedlings for a week. Seeds of a DGAT1::LUC plant were treated with EMS, and M2 seedlings which showed low LUC expression were further grown to obtain seeds. We then monitored bioluminescence of 32 to 48 individual M3 offsprings from each candidate. After screening ca 20,000 M2 seedlings, we obtained dozens of lines that consistently exhibit low LUC expression in M3 and at least two of them showed reduced LUC expression in developing seeds and reduced seed oil content. We also screened ca 10,000 T1 DGAT1::LUC plantsfurther transformed with enhancer T-DNA. Overexpression of one of a gene tagged with the enhancer under the 35S promoter caused enhanced LUC expression in seeds and increased oil content per seed. P13-036: METABOLIC RESPONSES OF QUERCUS SU- BER L. SEEDLINGS UNDER COMBINED ZINC AND DROUGHT STRESS Disante, K. 1 * - Fuentes, D. 3 - Cortina, J. 1 - Ljung, K. 2 - Lundgren, K. 2 1 Dept. de Ecología IMEM, Universidad de Alicante 2 Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences 3 Fundación Centro de Estudios Ambientales del Mediterráneo CEAM, Universidad de Alicante *Corresponding author, e-mail: kb.disante@ua.es Zinc (Zn) is an essential nutrient for higher plants that becomes toxic as its availability increases. In nature, stress factors seldom occur independently, but information on the interaction between Zn toxicity and other sources of stress is scarce. We have assessed the combined effect of Zn supply and low water availability on the metabolic profile of Quercus suber seedlings. The simultaneous application of two sources of stress triggered a strong metabolic response to maintain homeostasis. Seedlings were treated with four levels of Zn, ranging from 3 to 150 μM, while exposed to two contrasting irrigation regimes. Foliar and root Zn concentration increased with Zn availability, the increase being dependent on irrigation regime. The concentration of sugars, such as rhamnose and glucose, increased with Zn dose suggesting that these compounds may function as osmo-protectants. Observed differences in the organic acid profile (malic, aspartate and citric acid) in response to increasing Zn concentration may be related to tolerance. The relative concentration of α-Tocopherol, an antioxidant that deactivates photosynthesis-derived reactive oxygen species, increased at higher doses of Zn in well watered seedlings, but not in water-stressed seedlings. Low water availability resulted in an increase in the concentration of quinic acid and glucose, which may be related to protection against heavy metal and osmotic adjustment, respectively. In this study we relate metabolic changes with the physiological performance of Quercus suber seedlings under combined Zn and drought stress. Our results provide no evidence of synergism or antagonism between the two sources of stress. P13-037: BIOCHEMICAL AND PHYSICOCHEMICAL IMPLICATIONS OF THE REVERSIBLE STARCH PHOS- PHORYLATION Hejazi, M. - Fettke, J. - Steup, M. University of Potsdam AG Plant Physiology The phosphorylation of starch is mediated by two dikinases that esterify glucosyl residues at the C6 and the C3 position. Deficiency in either of the two dikinases indicates that the transient phosphorylation of starch is essential for an efficient turn-over. Arabidopsis chloroplasts contain at least one dual specificity phosphatase (DSP) that counteracts the two dikinases. This phosphatase (designated as SEX4) is highly homogenous to the laforin of the mammalian system. Non-functional laforin is associated with a major neurodegenerative disease, epilepsy. Arabidopsis mutants lacking a functional SEX4 possess a starchexcess phenotype and accumulate phosphorylated maltodextrins. The reversible phosphorylation of starch-like α-glucans has been studied by using crystalline maltodextrins representing either the A- or the B-type allomorph. Phosphorylation has been quantified by monitoring the incorporation of labelled phosphate from [β-33P]ATP into α-glucans. Dikinases possess a highly conserved pattern of amino acids. For a more detailed analysis of the reversible starch phosphorylation, a series of mutated dikinases has been generated in which single amino acids are exchanged. Catalytic features of the mutated dikinases have been studied both in the presence and in the absence of target glucans. The data obtained indicate an unexpected complexity of the biochemical action of the dikinases. P13-038: ALLELOPATHIC POTENTIAL OF THREE NE- PETA SPECIES: THE EFFECT OF NEPETALACTONE STEREISOMERS ON SEED GERMINATION AND AN- TIOXIDATIVE CAPACITY OF LEPIDIUM SATIVUM Nestorovic, J. 1 * - Misic, D. 1 - Siler, B. 1 - Zivkovic, S. 1 - Stojic, A.2 - Perisic, M. 2 - Grubisic, D. 1 1 Institute for Biological research “Sinisa Stankovic” 2 Institute of Physics *Corresponding author, e-mail: jakica81@gmail.com Allelopathic potential of three endemic Nepeta species, differing in their qualitative nepetalactone content, on the germination of Lepidium sativum L. seeds, was examined under in vitro conditions. Seeds of L. sativum were cocultivated with shoots of N. rtanjensis, N. sibirica and N. nervosa. The analyses of qualitative and quantitative nepetalactone content in the methanol extracts of in vitro grown shoots were performed by HPLC-DAD analysis. Trans-cis-nepetalactone was detected in shoots of N. rtanjensis Diklić & Milojević, while cis,trans-nepetalactone stereoisomer was present in N. sibirica L. No nepetalactone was observed in shoots of N. nervosa Royle & Bentham. The concentrations of nepetalactone in the atmosphere of culture vessels were detected by PTR-MS. The inhibition of L. sativum seed germination, and subsequently of seedling growth, was pronounced during cocultivation with N. sibirica, and especially N. rtanjensis. The increase in nepetalactone content in the atmosphere of glass jars was strongly correlated with the decrease of L. sativum seed germination and seedling growth. No significant inhibition in seed germination was observed for seeds cocultured with shoots of N. nervosa. The content of total phenolics, as well as the total antioxidative capacity of L. sativum seedlings, increased with increasing nepetalactone concentration in the atmosphere of glass jars. Our results suggest that nepetalactone possess strong allelopathic potential, and indicate that trans,cis-nepetalactone is more active than cis,transnepetalactone. P
FESPB 2010 - XVII Congress of the Federation of European Societies of Plant Biology P13-039: ENZYME AND METABOLITE PATTERN DU- RING POST-HARVEST STORAGE CONDITIONS IN BRASSICA RAPA L. CV. SYLVESTRIS Massaro, G. - Annunziata, M.G. - Iannuzzi, F - Nacca, F. - Carillo, P. - Fuggi, A.* Dipartimento di Scienze della Vita, Seconda Università di Napoli, Caserta *Corresponding author, e-mail: amodio.fuggi@unina2.it Keywords: broccoli, Brassica rapa, peroxidase, ascorbate, chlorophyll Vegetables after harvesting activate defence and senescent processes that affects also nutrtional, nutraceutic and organoleptic properties. They involve tissue wilting, yellowing and browning as well as the appearance of unpleasant tastes. In particular, chlorophyll degradation causing leaf yellowing and tissue browning are mainly due peroxidases and polyphenoloxidases. In this work peroxidases from Brassica rapa cv. sylvestris have been studied. Their distribution has been determined in different storage conditions (10 °C, 4 °C in air and 4 °C in a modified atmosphere) up to 20 days from the harvest. The florets, stem and leaf blade were analysed separately. Peroxidase activities increased during post-harvest, mainly in plants stored at 10 °C in air. The isoenzyme distribution was different in the analysed tissues and also changed differently in the storage time. The pattern of chlorophyll and carotenoids, as well as of ascorbate, inhibitor of some peroxidase isoenzyme, was also followed. As strong antioxidant they were also important nutraceutics. In addition the changes of metabolites, like carbohydrates, proteins, free amino acids, glucosinolates, nitrate and nitrite were also determined. Financial support was obtained by “Seconda Università degli Studi di Napoli”, “Ministero dell‘Università” and “Ricerca Scientifica e tecnologica” of Italy (PRIN 2006077008_005; 2008S9T3KK_003).
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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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