Ammonium uptake, transport and nitrogen economy in forest trees

AMMONIUM UPTAKE, TRANSPORT AND NITROGEN ECONOMY IN FOREST TREES Francisco M. Cánovas, Concepción Avila, Fernando N. de la Torre, Rafael A. Cañas, Belén Pascual, Vanessa Castro- Rodríguez, Jorge El-Azaz Departamento de Biología Molecular y Bioquímica, Facultad de Ciencias, Universidad de Málaga, Spain. Email: canovas@uma.es Forests ecosystems play a fundamental role in the regulation of global carbon fixation and preservation of biodiversity. Forest trees are also of great economic value because they provide a wide range of products of commercial interest, including wood, pulp, biomass and important secondary metabolites. The productivity of most forest ecosystems is limited by low nitrogen availability and woody perennials have developed adaptation mechanisms, such as ectomycorrhizal associations, to increase the efficiency of N acquisition and metabolic assimilation. The efficient acquisition, assimilation and economy of nitrogen are of special importance in trees that must cope with seasonal periods of growth and dormancy over many years. In fact, the ability to accumulate nitrogen reserves and to recycle N is crucial to determine the growth and production of forest biomass. Ammonium is the predominant form of inorganic nitrogen in the soil of temperate forests and many research efforts are addressed to study the regulation of ammonium acquisition, assimilation and internal recycling for the biosynthesis of amino acids, particularly those relevant for nitrogen storage. In our laboratory, we are interested in studying nitrogen metabolism and its regulation in maritime pine (Pinus pinaster L. Aiton), a conifer species of great ecological and economic importance in Europe and for which whole-transcriptome resources are available. The metabolism of phenylalanine plays a central role in the channeling of carbon from photosynthesis to the biosynthesis of phenylpropanoids and the regulation of this pathway is of broad significance for nitrogen economy of maritime pine. We are currently exploring the molecular properties and regulation of genes involved in the biosynthesis and metabolic fates of phenylalanine in maritime pine. An overview of this research programme will be presented and discussed. Research supported by Spanish Ministry of Economy and Competitiveness and Junta de Andalucía (Grants BIO2015-69285-R, BIO2012-0474 and research group BIO-114).

NAC-MYB basedtranscriptional networkinvolved in the regulation of phenylalanine biosynthesis in P. pinaster

P2-2 NAC-MYB-BASED TRANSCRIPCIONAL NETWORK INVOLVED IN THE REGULATION OF PHENYLALANINE BIOSYNTHESIS IN P. PINASTER Mª Belén Pascual, Rafael A. Cañas, Blanca Craven-Bartle, Francisco M. Cánovas and Concepción Ávila Departamento de Biología Molecular y Bioquímica. Facultad de Ciencias. Universidad de Málaga. Campus de teatinos s/n, Málaga, Spain Email: cavila@uma.es Conifer trees divert large quantities of carbon into the biosynthesis of phenylpropanoids, particularly to generate lignin, an important constituent of wood. Since phenylalanine is the precursor for phenylpropanoid biosynthesis, the precise regulation of phenylalanine synthesis and use should occur simultaneously. This crucial pathway is finely regulated primarily at the transcriptional level. Transcriptome analyses indicate that the transcription factors (TFs) preferentially expressed during wood formation in plants belong to the MYB and NAC families. Craven-Bartle et al. (2013) have shown that Myb8 is a candidate regulator of key genes in phenylalanine biosynthesis involved in the supply of the phenylpropane carbon skeleton necessary for lignin biosynthesis. This TF is able to bind AC elements present in the promoter regions of these genes to activate transcription. In Arabidopsis, the transcriptional network controlling secondary cell wall involves NAC-domain regulators operating upstream Myb transcription factors. We have identified in the P. pinaster genome three NAC proteins as potential candidates to be involved in vascular development. One of them, PpNAC1 is expressed both in xylem and compression wood from adult trees and has been thoroughly characterized. Its role upstream the transcriptional network involving Myb8 will be discussed. The understanding of the transcriptional regulatory network associated to phenylpropanoids and lignin biosynthesis in conifers is crucial for future applications in tree improvement and sustainable forest management.

Metabolic channelingof phe for lignin biosynthesis in maritime pine

METABOLIC CHANNELING OF PHE FOR LIGNIN BIOSYNTHESIS IN MARITIME PINE Jorge El-Azaz, Fernando de la Torre, Belén Pascual, Concepción Ávila and Francisco M. Cánovas Departamento de Biología Molecular y Bioquímica, Universidad de Málaga. Málaga, Spain Email: jelazaz@alu.uma.es The amino acid phenylalanine (Phe) is the main precursor of phenylpropanoids biosynthesis in plants. This vast family of Phederived compounds can represent up to 30% of captured photosynthetic carbon, playing essential roles in plants such as cell wall components, defense molecules, pigments and flavors. In addition to its physiological importance, phenylpropanoids and particularly lignin, a component of wood, are targets in plant biotechnology. The arogenate pathway has been proposed as the main pathway for Phe biosynthesis in plants (Maeda et al., 2010). The final step in Phe biosynthesis, catalyzed by the enzyme arogenate dehydratase (ADT), has been considered as a key regulatory point in Phe biosynthesis, due to its key branch position in the pathway, the multiple isoenzymes identified in plants and the existence of a feedback inhibition mechanism by Phe. So far, the regulatory mechanisms underlying ADT genes expression have been poorly characterized, although a strong regulation of the Phe metabolic flux should be expected depending on its alternative use for protein biosynthesis versus phenylpropanoid biosynthesis. This second fate involves a massive carbon flux compared to the first one. In this study we report our current research activities in the transcriptional regulation of ADT genes by MYB transcription factors in the conifer Pinus pinaster (maritime pine). The conifers channels massive amounts of photosynthetic carbon for phenylpropanoid biosynthesis during wood formation. We have identified the complete ADT gene family in maritime pine (El-Azaz et al., 2016) and a set of ADT isoforms specifically related with the lignification process. The potential control of transcription factors previously reported as key regulators in pine wood formation (Craven-Bartle et al., 2013) will be presented. Maeda et al. (2010) Plant Cell 22: 832-849. El-Azaz et al. (2016) The Plant Jounal. Accepted article, doi: 10.1111/tpj.13195 Craven-Bartle et al. (2013). The Plant Journal 74(5):755-766

Study of the impact federated erubrics on assessing competences in the practicum

Presentation Research of the Practicum and externships has a long history and involves important aspects for analysis. For example, the recent changes taking place in university grades allot more credits to the Practicum course in all grades, and the Company-University collaboration has exposed the need to study in new learning environments. The rise of ICT practices like ePortfolios, which require technological solutions and methods supported by experimentation, study and research, require particular examination due to the dynamic momentum of technological innovation. Tutoring the Practicum and externships requires remote monitoring and communication using ePortfolios, and competence-based assessment and students’ requirement to provide evidence of learning require the best tutoring methods available with ePortfolios. Among the elements of ePortfolios, eRubrics emerge as a tool for design, communication and competence-assessment. This project aims to consolidate a research line on eRubrics, already undertaken by another project -I+D+i [EDU2010-15432]- in order to expand the network of researchers and Centres of Excellence in Spain and other countries: Harvard University in USA, University of Cologne in Germany, University of Colima in Mexico, Federal University of Parana, University of Santa Catarina in Brasil, and Stockholm University in Sweden(1). This new project [EDU2013-41974-P](2) examines the impact of eRubrics on tutoring and on assessing the Practicum course and externships. Through technology, distance tutoring grants an extra dimension to human communication. New forms of teaching with technological mediation are on the rise and are highly valuable, not only for formal education but especially in both public and private sectors of non-formal education, such as occupational training, unemployed education and public servant training. Objectives Obj. 1. To analyse models of technology used in assessing learning in the Practicum of all grades at Spanish Faculties of Education. Obj. 2. To study models of learning assessment measured by eRubrics in the Practicum. Obj. 3. To analyse communication through eRubrics between students and their tutors at university and practice centres, focusing on students’ understanding of competences and evidences to be assessed in the Practicum. Obj. 4. To design assessment services and products, in order to federate companies and practice centres with training institutions. Among many other features, it has the following functions CoRubric(3) 1. The possibility to assess people, products or services by using rubrics. 2. Ipsative assessment. 3. Designing fully flexible rubrics. 4. Drafting reports and exporting results from eRubrics in a project. 5. Students and teachers talk about the evaluation and application of the criteria Methodology, Methods, Research Instruments or Sources Used The project will use techniques to collect and analyse data from two methodological approaches: 1. In order to meet the first objective, we suggest an initial exploratory descriptive study (Buendía Eisman, Colás Bravo & Hernández Pina, 1998), which involves conducting interviews with Practicum coordinators from all educational grades across Spain, as well as analysing the contents of the teaching guides used in all educational grades across Spain. 55 academic managers were interviewed from about 10 faculties of education in public universities in Spain (20%), and course guides 376 universities from 36 public institutions in Spain (72%) are analyzed. 2. In order to satisfy the second objective, 7 universities have been selected to implement the project two instruments aimed at tutors practice centers and tutors of the faculty. All instruments for collecting data were validated by experts using the Delphi method. The selection of experts had three aspects: years of professional experience, number and quality of publications in the field (Practicum, Educational Technology and Teacher Training), and self-rating of their knowledge. The resulting data was calculated using the Coefficient of Competence (Kcomp) (Martínez, Zúñiga, Sala & Meléndez, 2012). Results in all cases showed an average experience of more than 0.09 points. The two instruments of the first objective were validated during the first half of 2014-15 year, data collected during the second half. And the second objective during the first half of 2015-16 year and data collection for the second half. The set of four instruments (two for each objective 1 and 2) have the same dimensions as each of the sources (Coordinators, course guides, tutors of practice centers and faculty) as they were: a. Institution-Organization, b. Nature of internships, c. Relationship between agents, d. Management Practicum, e. Assessment. F. Technological support, g. Training and h. Assessment Ethics. Conclusions, Expected Outcomes or Findings The first results respond to Objective 1, where we find different conclusions depending on each of the six dimensions. In the case of internal regulations governing the organization and structure of the practicum, we note that most traditional degrees (Elementary and Primary grades) share common internal rules, in particular development methodology and criteria against other grades (Pedagogy and Social Education ). It is also true that the centers of practices in last cases are very different from each other and can be a public institution, a school, a company, a museum, etc. The memory with a 56.34% and 43.67% daily activities are more demands on students in all degrees, Lesson plans 28.18% 19.72% Portfolio 26.7% Didactic units and Others 32,4%. The technical support has been mainly used the platform of the University 47.89% and 57.75% Email, followed by other services and tools 9.86% and rubric platforms 1.41%. The assessment criteria are divided between formal aspects of 12.38%, Written expresión 12.38%, treatment of the subject 14.45%, methodological rigor of work 10.32%, and Level of argument Clarity and relevance of conclusions 10.32%. In general terms, we could say that there is a trend and debate between formative assessment against a accreditation. It has not yet had sufficient time to further study and confront other dimensions and sources of information. We hope to provide more analysis and conclusions in the conference date.