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P17 Plant Microbe Interaction P17-001: INFLUENCE OF MYCORRHIZAL FUNGI ON GROWTH, CHLOROPHYLL CONTENT AND K AND MG UPTAKE IN MAIZE UNDER DIFFERENT CONCENTRA- TIONS OF POTASSIUM AND MAGNESIUM Khanpour Ardestani, N.* - Reyhaneh, H. - Hassan Z.M. Tarbiat Modares University *Corresponding author, e-mail: n.khanpour@modares.ac.ir Mycorrhizal fungi have a positive effect on growth and nutrition of host plant. In this research Influence of Vesicular-Arbuscular Mycorrhiza) VAM) on Growth, chlorophyll Content and K and Mg Uptake in Maize in pot culture were studied.This experiment was performed using natural soil containing spores of Glomus spp. Mycorrhizal spores were exposed to 4 concentrations of K solution, i. e. 240 (soil K content), 360 and 480 mg/L and 3 concentrations of Mg, i.e. 4.8 (soil Mg content), 7.2 and 9.6 meq/L concurrently. Plants were watered every 4 days for 16 days with 50 ml distilled water. A pot with sterilized soil was used as negative control. For study of mycorrhizal colonization, very thin longitudinal sections of plant roots (>1mm in diam.) were prepared manually and were stained with lactophenol-cottonblue. Mycorrhizal percentage was determined by the grid-line intersect method. Sampling from root and shoot of maize had done. The results have shown that mycorrhizal plant significantly had higher dry and fresh weight and chlorophyll content than non-mycorrhizal control plants (p £ 0.05). The concentrations 7.2 meq/L of Magnesium and 360 mg/L of Potassium concurrently with 7.2 meq/L of Mg had better effect on morphological characters (Dry and fresh weight of root and shoot). Mycorrhizal colonization increased Mg uptake but decreased K uptake. Keywords: Magnesium and Potassium uptake, Growth, maize, Vesicular- Arbuscular Mycorrhiza ( VAM ) P17-002: CHARACTERIZATION OF AN ALTERNARIA ALTERNATA EXTRACELLULAR LACCASE INVOLVED IN THE PATHOGENESIS OF CITRUS FRUIT Del Río, J.* - Díaz, L. - Ortuño, A. - Pérez-Gilabert, M. Universidad de Murcia *Corresponding author, e-mail: jadelrio@um.es Fungi of the genus Alternaria are respondible for substantial preharvest losses in Citrus. The susceptibility to Alternaria alternata depends on the Citrus species (1, 2). In this communication, an extracellular laccase from A. alternata, apparently involved in the infection process in several Citrus species, is described for the first time. The enzyme was isolated from the culture medium of this fungus and characterized using ABTS as substrate. It exhibits a pH optimum of 3.5 and a Km of 1.9 mM. The effect of temperature was also studied. The enzyme is active between 15 and 45ºC, the optimum temperature being around 35ºC. The thermostability of the enzyme at its optimum temperature is very low and 50% of the activity is lost in 35 min. Different inhibitor agents were also studied, with L-cysteine and caffeic acid found to be the most effective. Study of the substrate specificity of this enzyme reveals that Citrus flavonoids are substrates of A.alternata laccase. These results suggest that the enzyme could P - Posters be involved in the degradation of flavonoids, observed in fruits infected with A. alternata. On the other hand, an increase of laccase activity was observed in the presence of Citrus fruit extracts. The possibility of considering laccase as a target to control the infection by Alternaria is also discussed. (1)Peever et al. (2000). Phytopathology.90, 407-414. (2) Ortuño et al. (2008). Physiological and Molecular Plant Pathology. 72, 162-166. Acknowledgments: This work was funded by the Projects AGR/11621/08 of the Consejería de Agricultura, Agua y Medio Ambiente de la Región de Murcia, Spain, and 08676/PI/08 of the Consejería de Educación, Ciencia e Investigación de la Región de Murcia-Fundación Séneca, Spain. P17-003: HOMOLOGY-DRIVEN PROTEOMICS RE- VEALS THE PROTEOME COMPLEXITY OF RESTING SPORES OF THE CLUBROOT PATHOGEN PLASMO- DIOPHORA Ludwig-Müller, J. 1 * - Knaust, A. 2 - Shevchenko, A. 2 1 Technische Universität Dresden 2 MPI-CBG, Dresden *Corresponding author, e-mail: Jutta.Ludwig-Mueller@tu-dresden.de The clubroot disease is one of the most damaging diseases of the Brassicaceae, which is caused by the protist Plasmodiophora brassica. The study of the pathogen is hampered due to its obligate biotrophic nature and the organism does not belong unequivocally into a systematic group, even within the protists. P. brassicae forms a group with three other plasmodiophorids whose genomes are uncharacterized. Sequence databases contain 50 proteins and 96 ESTs of P. brassicae having low homology to other organisms. We employed a proteomics approach to characterize several stages of the life cycle of the pathogen outside and inside the host plant Arabidopsis thaliana. Protein extracts of the pathogen were separated on 1D SDS PAGE and the entire lane was cut into 9 slices that were separately digested with trypsin and analyzed by LC-MS/MS. Proteins were identified by MASCOT searches, while unmathed spectra were interpreted de novo and candidate sequences submitted to homology-based identifications by MS BLAST. We demonstrated that resting spores are not dormant, but contain many proteins, of which 20 proteins were identified with 120 unique peptides matched: 2 hits were P. brassicae ORFs, 2 hits were the known P. brassicae proteins polyubiquitins 1 and 2, as well as 16 cross-species hits to proteins from other pathogens and fungi. Among these were house-keeping proteins as well as proteins for which a function in the life cycle can be predicted. In conclusion, we were able to extract protein material from resting spores and identify a number of proteins with no genome of the pathogen available. Homology driven proteomics altogether matched more peptides, validated cross-species MAS- COT hits and provided new protein identifications. P17-004: THE STUDY OF BIOCHEMICAL FEATURES OF PATHOGEN-DEPENDENT ENZYMES (CA2+ - ATPASE) UNDER INFECTION OF POTATO WITH FUSARIUM SO- LANI Manadilova, A.* - Tuleeva, G.T. - Duzgenbekova, B. Institute of Molecular Biology and Biochemistry *Corresponding author, e-mail: man_Alija@mail.ru The important part of the defensive response at infection is the activation of pathogene-dependent enzymes, including ATPase. The plasma membrane fractions,have been isolated from the potato suspension cells (Tamasha, Santa) different in their resistance to the Fusarium Solani. Main structure-functional parameters of Ca2+ ATPase of potato under normal and infected conditions were studied; the optimums for ion concentration of Ca2+, pH incubation medium and kinetic characteristics were determined. The influence of Fusarium Solani metabolites over the activity of enzyme was studied. After the infection by the fungus cultural fil- P
FESPB 2010 - XVII Congress of the Federation of European Societies of Plant Biology trate there is an increase of Ca2+ ATPase activity,but in resistant Tamasha variety, the enzyme activation is observed as quickly as one hour after infection, while the sensitive variety show the activation in 24 hours time. The infection leads to the change of physical and chemical parameters of the enzyme. It is seen from the changes of kinetic parameters; there is an increase of Km in infected potato cells. It is determined that carbohydrate and carbohydrate-lipidic fractions of Fusarium Solani mycelium are the suppressors of Ca2+ ATPase; protein fraction activates the enzyme. Naphthasarine, isolated from fungus mycelium by column chromatography, 2-3 times increases the activity of ATPase and identical to fusicoccin in terms of mechanism of action. P17-005: FACTORS AFFECTING FUMONISIN BIOSYN- THESIS IN FUSARIUM VERTICILLIOIDES: A STUDY ON FUM1 EXPRESSION UNDER IN VITRO CONDI- TIONS AND DURING EARLY STEPS OF MAIZE COLO- NIZATION. Visentin, I.* - Montis, V. - Valentino, D. - Tamietti, G. - Cardinale, F. Department of Arboriculture, University of Turin *Corresponding author, e-mail: ivan.visentin@unito.it Fumonisins are a family of mycotoxins (produced by Gibberella moniliformis, anamorph Fusarium verticillioides) that contaminate maize and maize-based products and cause great concern for human and animal health. The real role played by fumonisins in pathogenesis has been long controversial, although it is clear that they are phytotoxic. A better comprehension of the molecular mechanisms regulating their production could help clearing this point and preventing kernel contamination. In all eukaryotic organisms, acetylation of core histones and DNA methylation degree play a key role in the regulation of transcription, along the promoter regions. To assess the possibility that these epigenetic factors may be linked to fumonisin production, we observed the expression of a key biosynthetic gene (FUM1) in several strains of F. verticillioides after treatment with a histone deacetylase inhibitor, and investigated the methylation of FUM1 promoter under fumonisin-inducing and non-inducing conditions. Moreover we compared the activity of the endogenous FUM1 promoter (pFUM1) with that of the same region driving the ectopic expression of a transgene (GFP) after random insertion in other points of the genome in transgenic F. verticillioides strains (pFUM1::GFP). These are being tested for pFUM1 activity upon maize infection and in fumonisin-inducing and non-inducing conditions, to assess whether the FUM1 and GFP transcripts accumulate differently in dependence of the position of the corresponding coding and regulatory regions in the genome. Analysis of the data obtained from these assays could confirm the role of the epigenetic regulation in the fumonisin biosynthesis as already described for several other filamentous fungi. P17-006: EXPANSINS ARE INVOLVED IN CONTRO- LLING ROOT GALL SIZE AFTER INFECTION OF ARA- BIDOPSIS THALIANA WITH THE PROTIST PLASMO- DIOPHORA BRASSICAE Bretschneider, S.* - Wieczorek, K. - Grundler, F. - Ludwig-Müller, J. Institute of Botany, Technical University of Dresden *Corresponding author, e-mail: sabine.bretschneider@gmx.de Expansins are proteins that induce cell enlargement by loosening the matrix of the cell wall. They are regulated by symbionts and various pathogens as well as by auxin and mediate cell elongation via ‗acid-growth‘. Expansins are common in all land plants – from moss to flowering plants. In Arabidopsis there are 36 members of the expansin gene family (AtExp), in rice there are even 58. In this study we analysed whether expansins are specifically regulated during the clubroot disease of Arabidopsis thaliana. This disease is caused by the protist Plasmodiophora brassicae and affects only Brassicaceae. Symptoms are galls on the roots as well as wilted and stunted shoots. In a semi-quantitative RT-PCR expression analysis it was shown that most expansins are strongly up- or down-regulated in infected plants compared to healthy controls. In several promoter::GUS lines a direct correlation of expansin expression and infection structures was shown. Homozygous T-DNA mutants of AtExpA10 and AtExpA12 are more tolerant towards P. brassicae infection than wild type plants. Other possible candidates to be involved in clubroot formation are AtExpA1 and AtExpA15, based on expression studies, but this needs to be further examined. These studies potentially contribute to the development of highly tolerant or even resistant plants against P. brassicae infection. P17-007: THE EFFECT OF FUSARIUM SOLANI INFEC- TION ON ANTIOXIDANT ENZYMES IN POTATO (SO- LANUM TUBEROSUM) TUBERS Sapko, O. A. - Utarbayeva, A.* - Zhabayeva, DB. - Makulbek S, Baikara, B. - Kunayeva, R. M. M.A. Aitkhozhin Institute of Molecular Biology and Biochemistry *Corresponding author, e-mail: a.utar@mail.ru Plant have evolved complex of regulatory mechanisms in adapting to various environmental stresses. The oxidative stress is universal component of plant cell response reactions by pathogenesis. The antioxidant enzymes are effective components of protection to damage effects the reactive oxygen species. The fungus Fusarium solani cause the dry rot of potato tubers. The disease advance in tubers during storage. Changes in activities of ascorbate peroxidase (APX), catalase (CAT), superoxide dismutase (SOD) and peroxidase (PO) were investigated in potato tubers. The potato with different tolerance (resistant and sensitive) and pathogenic and nonpathogenic strains of F.solani was used. Character and intensity of enzyme response depended on initial tolerance of analyzed tubers and pathogenicity of fungus. The infection has caused fast (in 3 hours) 12-15-times increase CAT activity in tubers of sensitive cultivar and decrease in initial activity (on 30-70 %) in tuber of resistant cultivar. The higher level of enzymes activity to effect of pathogenic strain was observed. The maximal induction SOD, APX and PO activities registered in 24 hours. Higher level of activity SOD and PO was registered in resistant cultivar infected pathogenic strain of fungus. Activity APX increased in sensitive tubers infected nonpathogenic stain. The received results confirm the important role of antioxidant enzymes in interaction F.solani - S.tuberosum. P17-008:L-2-OXOTHIAZOLIDINE-4-CARBOXYLIC ACID (OTC) PARTIALLY PROTECTS PEACH PLANTS FROM SHARKA DISEASE: EFFECT IN ANTIOXIDATI- VE METABOLISM AND PROTEIN EXPRESSION. Hernandez, J.* - Clemente-Moreno, M.J. - Diaz-Vivancos, P. - Barba-Espín, G. - Martínez-Gómez, P. Grupo de Biotecnología de Frutales CEBAS-CSIC *Corresponding author, e-mail: jahernan@cebas.csic.es In this work, we investigate the effect of L-2-oxothiazolidine-4- carboxylic acid (OTC) pre-treatments and their interaction with Plum pox virus (PPV, Sharka disease) infection on the antioxidative metabolism of “GF305” peach (Prunus persica L. Batsch) at subcellular level. Experiments were carried out during 4 years (2007 to 2010) in controlled greenhouse conditions. In general, OTC increased the length of peach plants and decreased the percentage of branches showing Sharka symptoms in their leaves. This partial protection obtained by OTC treatments, correlated with a lower PPV contents evaluated using an ELISA-DASI test, mainly during the second cycle of growth in each year. At the same time, these plants showed a higher redox state of glutathione as well as an increase in GSH-related enzymes (GPX, GST and G6PDH) and POX activity at subcellular level. Finally, the effect of OTC in the differential expression of proteins in healthy and PPV-infected leaves will be discussed.
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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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