Luca Espen Co-tutor Characterization of changes in root membrane proteomes in maize (Zea mays L.) in responses to nitrogen sources PhD School on Agriculture, Environment and Bioenergy (XXXV cycle, 2019-2021) Chiara Muratore PhD Student Bhakti Prinsi Tutor Luca Espen Co-Tutor
13
Embed
Characterization of changes in root membrane proteomes in ...
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
‘Nitrogen cycle’ (Lappalainen, H. K., et al., 2005)
RELEVANCE OF THE TOPIC AND STATE OF THE ART
Plants elaborate responses to enhance
nitrogen use efficiency such as uptake
activity and root architecture
Inorganic
(Nitrate, NO3- ;
Ammonium, NH4+)
Organic
(amino acids, peptides, urea)
Inorganic
fertilizers
Organic
fertilization,
biostimulants
THE ROLE OF ROOTS IN N-NUTRITION
WHY FOCUS ON MEMBRANE PROTEOMES?
➢ Perception of N sources
➢ Nutrient uptake
➢ Metabolism (respiration, protein synthesis …)
➢ Communication among organelles (transport,
storage, regulation …)
THE KEY ROLES OF ROOT IN N NUTRITION
➢ PERCEPTION
➢ ADPTATION
➢ SIGNALLING
Taiz et al., (2013)
Britto & Kronzucker, (2002) Forde, G., (2013)
Hordeum vulgare Arabidopsis thaliana L.
➢ How root perceives and adapt to different N-availabilities?
➢ How membrane proteome vary in response to different N-sources?
➢ What is the correlation at physiological and metabolic level?
AIMS OF THE PROJECT
Characterization of changes in
root membrane proteomes in
maize (Zea mays L.)
in responses to nitrogen sources
INTEGRATIVE APPROACH
➢ PHENOTIPICAL CHARACTERIZATION
➢ PHYSIOLOGICAL AND BIOCHEMICAL EVALUATIONS
➢ PROTEOMIC PROFILING
First year Second year Third year
▪ Literature review
▪ Focus on inorganic N-
sources
▪ Optimization of the
hydroponic nutritional
treatments and analytical
procedures for proteome
study
▪ Study of the root
responses to NO3- e
NH4+:
▪ Proteomics
▪ Phenotyping of RSA
▪ Physiology
▪ Focus on of N-organic
sources (amino acids)
▪ Study of the root
responses to amino acids
(Glu, Gln, biostimulants..):
▪ Proteomics
▪ Phenotyping of RSA
▪ Physiology
▪ Visiting PhD exchange
PROJECT SCHEDULE AND MAJOR STEPS
▪ Different timings (plant
growth, nutritional
treatments..)
▪ Validation of the results by
means of bioinformatic
tools, targeted analyses
and complementary
approaches
▪ Thesis write up
• Marschner H: Mineral Nutrition of Higher Plants London: Academic PressLimited; 1995.
• Britto, D. T., & Kronzucker, H. J. (2002). NH4+ toxicity in higher plants: acritical review. Journal of plant physiology, 159(6), 567-584.
• Miller, A. J., & Cramer, M. D. (2005). Root nitrogen acquisition andassimilation. In Root physiology: From gene to function (pp. 1-36).Springer, Dordrecht.
• Forde, B. G., & Lea, P. J. (2007). Glutamate in plants: metabolism,regulation, and signalling. Journal of experimental botany, 58(9), 2339-2358.
• Xu, G., Fan, X., & Miller, A. J. (2012). Plant nitrogen assimilation and useefficiency. Annual review of plant biology, 63, 153-182
• Andrews, M., Raven, J. A., & Lea, P. J. (2013). Do plants need nitrate? Themechanisms by which nitrogen form affects plants. Annals of appliedbiology, 163(2), 174-199.
• Forde, B. G., & Roberts, M. R. (2014). Glutamate receptor-like channels inplants: a role as amino acid sensors in plant defence?. F1000primereports, 6.
• Buchanan, B. B., Gruissem, W., & Jones, R. L. (Eds.). (2015). Biochemistryand molecular biology of plants. John Wiley & Sons.
• Kiba, T., & Krapp, A. (2016). Plant nitrogen acquisition under lowavailability: regulation of uptake and root architecture. Plant and CellPhysiology, 57(4), 707-714
• Prinsi, B., & Espen, L. (2018). Time-course of metabolic and proteomicresponses to different nitrate/ammonium availabilities in roots andleaves of maize. International journal of molecular sciences, 19(8), 2202.
…
• Hodges, T. K., & Mills, D. (1986). Isolation of the plasma membrane.
In Methods in Enzymology (Vol. 118, pp. 41-54). Academic Press.
• Larsson, C., Widell, S., & Kjellbom, P. (1987). [52] Preparation of high-
purity plasma membranes. In Methods in Enzymology (Vol. 148, pp. 558-
568). Academic Press.
• Alexandersson, E., Saalbach, G., Larsson, C., & Kjellbom, P. (2004).
Arabidopsis plasma membrane proteomics identifies components of
transport, signal transduction and membrane trafficking. Plant and
Cell Physiology, 45(11), 1543-1556.
• Zhang, Z. J., & Peck, S. C. (2011). Simplified enrichment of plasma
membrane proteins for proteomic analyses in Arabidopsis
thaliana. Proteomics, 11(9), 1780-1788.
• Wang, X., Bian, Y., Cheng, K., Zou, H., Sun, S. S. M., & He, J. X. (2012). A
comprehensive differential proteomic study of nitrate deprivation in
Arabidopsis reveals complex regulatory networks of plant nitrogen
responses. Journal of proteome research, 11(4), 2301-2315
• Takahashi, D., Kawamura, Y., & Uemura, M. (2013). Detergent-resistant
plasma membrane proteome to elucidate microdomain functions in
plant cells. Frontiers in plant science, 4, 27
• Yadeta, K., Elmore, J. M., & Coaker, G. (2013). Advancements in the
analysis of the Arabidopsis plasma membrane proteome. Frontiers in
plant science, 4, 86..
• Szymanski, W. G., Zauber, H., Erban, A., Gorka, M., Wu, X. N., & Schulze,
W. X. (2015). Cytoskeletal components define protein location to