Book of Abstracts - Geyseco

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P - Posters Ebadi M.¹* - Iranbakhsh, A.² ¹Islamic Azad University, Damghan Branch ²Islamic Azad University, Aliabad Katoul Branch *Corresponding author e-mail: iranbakhshar@yahoo.com In this study, potatos cultivated through mono-nodule explanation, were moved from shoot-formation induction media to MS induction media or the purpose of microtuberization of which the consistence/density of BAP (1, 2, 5 & 10 mg -1 ) and Sucrose (30, 40, 60 & 80 mg -1 ) were shifted in darkness. Although no microtuberization was observed in the induction media containing 3% Sucrose, the number of white branches sprouted from peripheral buds were enhanced. It is observed that increase in BAP consistence on one hand led to deacrease in the number of branches. In media containing 4% sucrose and with low BAP consistency (1 mgl -1 , 2 mgl -1 ) shoot-formation (branching) was not perceived. While BAP in the levels (5 mgl -1 to 10 mgl -1 ) was augmented in the induction media containing 4% sucrose, the growth of lateral buds accompanied by delayed microtuberization was four weeks after induction. In such media, the microtubers were created from the alteration to the growth pattern in sub-apical area with positive geotropism. In BAP 10 mgl -1 consistency, 50% of tubers were attached to the branches. In induction media containing 6% sucrose and BAP 1 & 2 mgl -1 , the microtubers were grown on peripheral branches until the end of week2. In such groups, no tubers bigger than 7 mm, was seen. In sucrose with 8% consistency, the highest percentage of microtuberization was perceived in BAP different consistencies. In BAP 10 mgl -1 consistency joined by 8% Sucrose, the maximal number of normal tubers attached to the pedicle/rhizome with larger dimension was formed. In such induction media, the formation of microtubers in the 1st week was commenced and the 2nd week was completed. The growth of microtubers continued up to the 7th week. The latency of microtubers was protracted of which the duration was about 30 months. P05-017: A GENETIC ENGINEERING APPROACHES TO INCREASE STARCH PRODUCTION IN PLANT Halil, I.* - Baris, I. – Tuncel, A. – Ozber, N. - Keskin, O. Koc University *Corresponding author e-mail: hkavakli@ku.edu.tr ADP-glucose pyrophosphorylase (AGPase), a key allosteric enzyme involved in higher plant starch biosynthesis, is composed of pairs of large (LS) and small subunits (SS). Current evidence indicates that the two subunit types play distinct roles in enzyme function. Recently the heterotetrameric structure of potato AGPase has been modeled. In the current study, we have applied computational tools and identified critical amino acids of the potato AGPase LS and SS subunits that interact with each other during the native heterotetrameric structure formation. We have further shown the role of the LS amino acids in subunit-subunit interaction by yeast two-hybrid, bacterial complementation assay and native gel. During ouranalysis we have found that lateral interaction of the LS-SS is much stronger than the longitudinal one, and it is mainly mediated by hydrophobic interactions. Also, we have utilized a reversion genetic approach to obtain stable heterotetrmeric AGPase using one of the LS AGPase mutant. Currently, we are characterizing the mutants in E.coli system. Thistudy will not only enhance our understanding of the interaction between the SS and the LS of AGPase, but will also enable us to engineer proteins to obtain better assembled variants of AGPase which can be used for the improvement of plant yield. P05-018: SOMATIC EMBRYOGENESIS IN THE GRAPE- VINE: SOMACLONAL VARIANTS AND DEVELOPMEN- TAL PHYSIOLOGY Rey, M.¹* - Domínguez, C¹ - Acanda, Y.¹ - Rey, L.¹ - González, M.V.² - Rodriguez, E.³ - Santos, C.³ - Centeno, M.L. 4 - Prado, M.J.¹ ¹Universidad de Vigo ²Universidad de Santiago de Compostela ³Universidad de Aveiro 4Universidad de León *Corresponding author e-mail: mrey@uvigo.es We have developed a complete protocol for plant regeneration through somatic embryogenesis for six autochthonous grapevine cultivars from Galicia (north-western Spain). Somatic embryogenesis was induced in anthers and ovaries collected all along the binucleate pollen microsporogenesis stage, indicating a wide window of competence for induction. The trueness-to-type of the somatic embryogenesis-regenerated plants was tested by flow cytometry and microsatellite analysis. Results showed that several somaclonal variants by ploidy level changes were obtained, including octoploid, tetraploid and mixoploid (diploid plus tetraploid cytotypes) plants. A particular case was that of the cultivar ‘Brancellao’ from which both tetraploid and diploid plants were regenerated. As we found that 50% of the adult field-grown mother ‘Brancellao’ plants analysed were mixoploid, this suggests that regenerated plants originated either from somaclonal variation or by separation of genotypically different cell layers through somatic embryogenesis. All somatic embryogenesis-regenerated plants were trueto-type according to the microsatellite genotypes, with the exception of six ‘Torrontés’ plants showing a mutant allele (231) instead of the normal one (237) at the locus VVMD5. Despite these results, deficiencies in the maturation of the somatic embryos reduce the efficiency of the somatic embryogenesis regeneration system. We are studying the role phytohormones like ABA and IAA are playing during somatic embryo development, to understand the physiological mechanisms for such deficiencies. Preliminary results are presented, with the aim to further contribute to increase the efficiency of the somatic embryogenesis process in the grapevine. P05-019: DEVELOPING DROUGHT AND BROOMRAPE RESISTANT SUNFLOWER GERMPLASM UTILIZING WILD HELIANTHUS SPECIES Petcu, E.* - Joita Pacureanu, M. National Agricultural Research and Development Institute, Fundulea, Romania *Corresponding author e-mail: petcue@ricic.ro The production of sunflower, the most important oil plant in Romania, is quantitatively and qualitatively affected by the extreme meteorological events during last time. Among climatic factors, drought and heat are the most important. Broomrape (Orobanche cumana) is a very harmful root parasitic weed of sunflower under dry area. The project aim is the increasing of sunflower adaptability to stresses by improving biological material tolerance to drought, heat and broomrape. For this, introgression of resistance genes from wild species was attempted, using embryo rescue techniques, combined with classical procedures to improve crossing, selfpollination and backcrossing. Interspecific F 3 hybrids were produced between six cultivated Romanian inbred sunflower line and wild species H. argophyllus and F 2 . hybrids between H. argophyllus and four cultivated inbred sunflower. Crosses between four cultivated inbred Romanian sunflower line and H. maximiliani were backcrossed with same cultivated inbred line advanced to BC 1 F 3 and BC 2 F 3 generation in 2008 and 2009 for seed increase. Replicated green house tests with several progeny were screened for drought and broomrape resistance. The results indicated good resistance, suggesting successful gene introgression. Identified resistant lines will retest in 2010 and the results will be used to release germplasm, providing new resistance genes to enhance drought and broomrape resistance in sunflower. P
FESPB 2010 - XVII Congress of the Federation of European Societies of Plant Biology P05-020: IN VITRO STRESS INDUCED BY THE PHO- MOPSIS HELIANTHI FILTRATE ON SOME PHYSIOLO- GICAL INDICES AND OIL QUANTITY AND QUALITY ON SUNFLOWER Florentina, S.* - Sauca, F. - Petcu, E. National Agricultural Research and Development Institute, Fundulea, Romania *Corresponding author e-mail: tina@ricic.ro Using in vitro screening, the goals of this study were to contribute to the knowledge regarding of the influence of stress induced by Phomopsis helianthi filtrate on some Romanian inbred lines and identification the inbred lines with high level of tolerance to the pathogen. For in vitro testing to Phomopsis helianthi pathogen a total of 14 Romanian inbreed lines were used. As follows of the treatment applied on MS culture medium supplemented with 150ml/l filtrate and on the basis of the results obtained regarding the leaf index, chlorophyll contend, TKW, seed oil percentage and its composition determination. Genotypes with increased resistance to this pathogen have been selected. The determination have been performed by the Minolta Chlorophyll meter (SPAD units) for chlorophyll contends, RMN methods for oil contend and gas-chromatography method (Shimadzu-GC-14B) for fatty acid contends from oil. The results obtained at gas chromatograph underline the fact that from the five fatty acids from sunflower oil, the oleic acid decreases after treatment in all genotypes, excepting the LC 4010 line. We positively notice the fact that the linolenic acid which reduces the oil stability, was detected only in three genotypes but in very small quantities. . Eight genotypes in which the leaf area was not diminished by the treatment as compared with control have been identified. As regards the chlorophyll content, all tgenotypes, at the treatment variant, the average / variant was diminished with 5.2 SPAD units vs. the control. In variant treated with filtrate, TKW was diminished at seven out of the 14 genotypes. The oleic acid content was more decreased in all lines excepting the LC 4010 line. Key words: Phomopsis helianthi; in vitro culture; chlorophyll contend; in vitro. P05-021: LIPID CHANGES IN CELL CULTURES OF SILYBUM MARIANUM IN RESPONSE TO ELICITA- TION BY METHYL JASMONATE Corchete, P.* - Cacho, M. Dep. Fisiología Vegetal. Universidad de Salamanca *Corresponding author e-mail: corchpu@usal.es The biosynthesis of silymarin, a medicinal compound of the fruits of Silybum marianum, is severely reduced in cell cultures. Methyl jasmonate (MeJA) elicits silymarin synthesis and its release to the culture medium. Recent work has shown that phospholipase D (PLD) and its product phosphatidic acid (PA) mediates silymarin secretion. To comprehensively determine the overall effects of MeJA on cellular membranes, glycerolipid composition of cell cultures and changes during elicitation were analyzed by ESI-MS/MS (Kansas Lipìdomics Centre, USA). With this methodology 152 molecular lipid species were identified. Phosphatidyl choline (PC) (42,46%), P ethanolamine (PE) (23,56%) and P inositol (PI) (22,52%), were the main glycerophospholipids; monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), in less proportion (2 and 3%), were also present in membranes. Acyl species were represented by insaturated C16 and C18 fatty acids. A quantitative decrease of PC and PE was observed after 24h of MeJA treatment; galactolipids or PI levels were not changed. Elicitation induced rapid and transient increases in LysoPC and LysoPE. PA also increased and its levels remained high up to 8h of elicitation. The results suggest that besides PLDs, PL As were also activated by MeJA. However, the low level of the PLA products indicates that PLD is more active than PLA in Silybum cultures. These findings show that elicitation causes a selective hydrolysis of membrane lipids, indicate which are the affected molecular species and inform that phospholipases and not galactolipases are involved in the process. In addition, the decrease in both PC and PE suggests that the two lipids are potential substrates for PLDs. Work was financed by MICINN (BFU-02876) Spain P05-022: OVER-EXPRESSION OF THE ARABIDOPSIS HISTONE ANTI-SILENCING FACTOR A, SGA1, IN- CREASES STABLE TRANSFORMATION OF PLANT CELLS Tenea, G.¹* - Spantzel, J. – Oltmanns, H.² - Gelvin, S.B.² ¹University of Bucharest, Institute of Genetics ²Department of Biological Sciences, Purdue University *Corresponding author e-mail: tenea.gabriela@gmail.com Agrobacterium-mediated genetic transformation is the dominant technology used for the production of genetically modified transgenic plants. Extensive research has been conducted which is aimed at understanding the role of host proteins, especially chromatin proteins, in the transformation process. A long-term goal is to manipulate the expression of these proteins to improve transformation. RNAtiargeted against 109 Arabidopsis chromatin genes demonstrated a role for many chromatin proteins, such as the anti-silencing factor SGA1, in Agrobacterium-mediated transformation. We investigated the effects of over-expressing a SGA1 cDNA on transgene expression and Agrobacterium-mediated transformation. Unlike the situation with many histone cDNAs, over-expression of a SGA1 cDNA did not increase transient expression of a gusA gene co-transfected into tobacco BY-2 protoplasts. Moreover, over-expression of SGA1 did not increase expression of a previously integrated gusA transgene. However, transgenic Arabidopsis plants containing additional copies of a SGA1 cDNA displayed increased accumulation of SGA1 mRNA and greater susceptibility to transformation. Moreover, DNA blot analysis showed an increase in T-DNA integration into the genome of Arabidopsis plants over-expressing SGA1. We speculate that SGA1 might lead to enhanced plant transformation by allowing T-DNA and complexed proteins greater access to plant target DNA, thus facilitating T-DNA integration. P05-023: PRODUCTION OF SOYBEAN GRAINS AND TO- BACCO SEEDS HAVING HIGHER METHIONINE CON- TENT Amir, R.¹* - Godo, I.¹ - Matityahu, I.¹ - Shikui, S.² - Wensheng, H.² - Tianfu, H.² ¹Migal, resreach institute ²Chinese Academy of Agricultural Sciences *Corresponding author e-mail: rachel@migal.org.il This study describes the first modification of methionine biosynthesis in seeds of tobacco, the model plant, andof soybean, the crop legume plant (Glycin max L.). Unlike vegetative tissues, the seed-specific expression of Arabidopsis in its unregulated deleted form of cystathionine gamma synthase (At D-CGS) in tobacco seeds led only to a slight accumulation of methionine. It was previously found that the seed-specific expression of feedback-insensitive bacterial aspartate kinase (AK), which operates up-stream to CGS in the methionine biosynthesis pathway, led to two-fold higher methionine content in transgenic seeds. This finding suggests that in tobacco seeds, the availability of the carbon/amino skeleton of methionine limits its synthesis and less the activity rate of CGS. To further test this assumption, plants’ seeds-specific expression of AtD-CGS were crossed with those expressing AK. The methionine level increased up to four-fold higher in seeds of progenies expressing both foreign genes compared to wild-type seeds, suggesting that indeed the carbon/amino skeleton limits methionine content in seeds. These results are encouraging us to try to elevate methionine content in
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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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