Multilevel analysis towards drought tolerance in Legumes

1. Multilevel analysis towards drought tolerance in Legumes [P23441-B20] Start: 2011 Stop: 2014 • PhDStudents: • Christiana Staudinger: • Plant Physiology, ProteomicsandMetabolomics • Vlora Mehmeti: • Plant PhysiologyandMetabolomics • David Lyon: • Proteomics,MetabolomicsandBioinformatics One of the most crucial functions of plant cells is their ability to respond to alterations in their environment. Understanding the connections between initial responses and the downstream events that constitute successful adjustment to its fluctuating environment is one of the challenges of plant biology research. Investigations on transcript level are the most common studies so far. Besides transcript analyses, new technologies based on mass spectrometry allow for the comprehensive study on metabolite and protein level. Past studies have focussed on different stresses such as temperature, drought or salt using various plants and technologies. All these data have improved the understanding on the complexity of the plant response depending on the intensity and duration of homeostatic perturbation. However, due to the diversity of the research data and experimental conditions a comparison and integration is difficult or even impossible. Thus to gain better insights and to be able to visualise the complexity of the plant respond, integrative analyses combining different technologies and standardised cultivation conditions are becoming necessary. Legumes are major sources of vegetable protein and indispensable for sustainable agriculture due to their ability to fix atmospheric nitrogen via their symbiosis with soil rhizobia. These bacteria colonize legume roots in specialized organs called nodules. However, a full understanding of the interactive regulatory mechanisms between plant and bacteroids towards increased stress tolerance is not accomplished so far. To contribute to a deeper understanding of responses of legumes to major constraints, the proposed project aims at employing comparative systems biology. Supported by bioinformatic modelling strategies, novel metabolic and proteomic key mechanisms that may serve as regulatory targets for improving stress-tolerance in legumes will be identify. • Guest Scientists • Erena Gil (UniversidadPublica de Navarra): • Proteomics; Medicago andSoybeannodules; drought stress. • Dr. M.AngelesCastillejo, Universidad de Cordoba (fundedbytheSpanishMinistryofScience and Innovation): • Plant PhysiologyandProteomics; 15N-metabolic labelling • Veronica Castañeda (UniversidadPublica de Navarra): • Proteomics; Medicago rootplasmamembranes; drought stress • PI: Dr. Stefanie Wienkoop Link to Publications • Bioinformaticsupervision: Dr. Volker Egelhofer Link toConferencesand Posters

2. Multilevel analysis towards drought tolerance in Legumes [P23441-B20] Publications: • Peer Rev. Journals: • - D. Lyon, M.A. Castillejo, C. Staudinger, W. Weckwerth, S.Wienkoop and V. Egelhofer (2014). Automated protein turnover calculations from 15N partial metabolic labeling LC/MS shotgun proteomics data. PlosOne, 9(4): e94692. doi: 10.1371/journal.pone.0094692. • - C. Staudinger, V. Mehmeti, R. Turetschek, D. Lyon, V. Egelhofer and S. Wienkoop (2012). • Possible role of nutritional priming for early salt and drought stress responses in Medicago truncatula. Frontiers Plant Proteomics. doi: 10.3389/fpls.2012.00285 • - A. Mari, D. Lyon, L. Fragner, P. Montoro, S. Piacente, S. Wienkoop, V. Egelhofer, W. Weckwerth (2012). Phytochemical • composition of Potentillaanserina L. analyzed by an integrative GC/MS and LC/MS metabolomics platform. Metabolomics Doi:10.1007/s11306-012-0473-x • Book chapters: • - M.A. Castillejo, C. Staudinger, V. Egelhoferand S. Wienkoop (2014). Medicago truncatula proteomicsforsystemsbiology: novel rapid shotgun LC-MS approachfor relative quantificationbased on Full-Scan Selective Peptide Extraction (Selpex). Chapter 22 In: Plant ProteomicsMethodsandProtocols. Ed. J.V. Jorrin Novo, S. Komatsu, W. Weckwerth, S. Wienkoop, : Springer New York. Software: http://promex.pph.univie.ac.at/protover/Publication: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3988089/ Download: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3527748/ Download: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3651535/ Abstract: http://www.ncbi.nlm.nih.gov/pubmed/24136531 severalpublications still in progress!

3. Multilevel analysis towards drought tolerance in Legumes [P23441-B20] Conferencesand Posters: 2013 17. – 21.3. Proteomics Formum 2013, Berlin, Germany a) D. Lyon, M.A. Castillejo, C. Staudinger, V. Mehmeti, V. Egelhofer and S. Wienkoop Protein turnover calculations from partial metabolic labeling in Plantaexperiments. b) Staudinger Christiana, Castillejo Maria-Angeles, Mehmeti Vlora, Lyon David, Egelhofer Volker, Wienkoop Stefanie Combined Label-free Quantification Techniques to Detect Water Deficit Induced Changes in the Medicago truncatula Proteome 8. – 13.6. 61stASMS Conference, Minneapolis, MN, USA D. Lyon, S. Wienkoop, V. Egelhofer An automated program for protein turnover calculations from LC/MS shotgun proteomics data resulting from partial metabolic labelling experiments. 4. - 10.08. 7th European Summer School “advanced proteomics”, KlosterNeustift, Brixen, South Tirol, Italy Staudinger C., Castellejo-Sanchez MA., Lyon D., Egelhofer V. and Wienkoop S. Suffer and recover: How Medicago truncatula acclimates to water deficit 2. - 6.09. Interdrought-IV, Crown Perth, Western Australia Staudinger C., Turetschek R., Mehmeti V., Castillejo M., Lyon D. and Wienkoop S. Legume nitrogen nutrition-related strategies to cope with drought Link Poster 1 Link Poster 2 Link Poster 3 Link Poster 4 Link Poster 5 2012 18.-21.2. ORAL PRESENTATION - VIPCA, Internat. Conf. Plant Growth, Nutrition & Environment Interactions, Vienna, Austria V. Mehmeti, S. Wienkoop Molecular Changes in the metabolome of Medicago truncatula Induced by Short Term Salt and Drought Stress. 22.-25.2. VIPCA, Intern. Conf. Plant Abiotoc Stress Tolerance IIC, Vienna, Austria. C. Staudinger, V. Mehmeti, E. Gil, E.M.Gonzalez, S. Wienkoop RhizobialSymbiosis-related Effects on Drought Tolerance in Medicago truncatula. 2.-5.09. 10th European Nitrogen Fixation Conference, Munich, Germany a) Staudinger Christiana, MehmetiVlora, Gil Erena, Gonzales Esther and Wienkoop Stefanie Proteomiccharacterisationofmicrosymbtiont-dependentchangesin Medicago truncatula roots and shoots. b) V. Mehmeti, C. Staudinger and S. Wienkoop DIFFERENCIAL NITROGEN SOURCE ASSOCIATED METABOLITE CHANGES OF MEDICAGO TRUNCATULA INDUCED BY SHORT TERM SALT AND MILD DROUGHT STRESS Link Poster 6 Link Poster 7 Link Poster 8