La genómica de las plantas

La mejora vegetal es una actividad tan antigua como la propia agricultura. El estudio de las variedades genéticas de la vegetación permite conocer en profundidad sus funciones y mecanismos para localizar los beneficios alimenticios, textiles, cosméticos o combustibles que la naturaleza ha puesto al servicio del ser humano

Nitrogen metabolism and gene expression landscape in maritime pine

This work was supported by the ProCoGen grant (FP7-KBBE-2011-5) and by the MicroNUpE grant (BIO2015-73512-JIN).Maritime pine (Pinus pinaster Aiton) is one the most important conifer species in the southwestern Mediterranean region because of its economic and environmental potential. For this reason, a work program in functional genomics has been developed in the frame of the ProCoGen project. One objective was to complete the knowledge about P. pinaster transcriptome with the tissue-specific localization of the gene expression of the low accumulated transcripts in sharper regions (Cañas et al. 2017). In order to reach these objectives total RNA was obtained from isolated tissues through laser capture microdissection (LCM). Due to the limiting amount from these extracts, the RNA samples were reverse-transcribed and the resultant cDNA amplified using our CRA+ protocol (Cañas et al. 2014). The obtained reads were assembled to improve the previous reference transcriptome. Reads were mapped against this transcriptome and the read accounts analyzed in order to found gene co-expression networks using the WGCNA software. These results have allowed us the characterization of nitrogen metabolism in maritime pine during the seedling stage stablishing relationships between the different components. This include the identification of new genes with low or very localized expression as occurred for the PpGS1c gene encoding a new cytosolic glutamine synthetase. From this starting point, we are developing a new project, MicroNUpE, to identify and study the genes involved in ammonium uptake and regulation in different root tissues that will be isolated through LCM. Cañas et al. (2017). Plant J, doi:10.1111/tpj.13617. Cañas et al. (2014). Tree Physiol, 34:1278-1288.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Molecular and functional approaches to enhance biomass production in forest trees

Forest trees comprise a large group of angiosperm and gymnosperm species that play a crucial role in global carbon fixation, and maintenance of biodiversity. Forest trees are also of great economic importance since they provide a wide range of products of commercial interest, including wood, pulp, biomass and important secondary metabolites. A sustainable management of forest resources is needed to preserve natural forests and to meet the increasing international demands in the production of wood and other forest-derived products. New advances and developments in biotechnology may contribute to accelerate the domestication of important traits for forest productivity. Nitrogen assimilation and recycling play a key role in tree growth and biomass production and we firmly believe that knowledge on nitrogen metabolism will lead to approaches aimed at increasing forest productivity. We are interested in studying nitrogen metabolism and its regulation in maritime pine (Pinus pinaster Aiton), a forest tree species of great economic and ecological importance in the Mediterranean area and a relevant model for conifer genomic research in Europe. An overview of our research programme will be presented and discussed. Research supported by Spanish Ministry of Economy and Competitiveness and Junta de Andalucía (Grants BIO2012-33797, PLE2009-0016 and research group BIO-114).Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Analysis of NPF and NRT transporter families regarding the nitrate nutrition in maritime pine (Pinus pinaster)

Nitrogen is an essential element for life and the main limiting nutrient for plant growth and development1. The main forms of inorganic nitrogen in soils are nitrate and ammonium, which relative abundances depend on environmental conditions such as temperature. In agricultural soils the most abundant nitrogen form is nitrate because the use of chemical fertilizers however in natural ecosystems nitrogen soil composition can be more complex. Conifers are tree gymnosperms with a wide distribution although their large forests dominate the boreal ecosystems where nitrification is limited and ammonium is the main nitrogen soil source2. In this context, conifers have an appreciable tolerance to ammonium. Maritime pine (Pinus pinaster Aiton) is a conifer from the western Mediterranean region of high economic and ecological interest in Spain, France and Portugal. This pine is also a research model tree with different genomic resources such as a reference transcriptome and a gene expression atlas3. Taking advantage of these resources the members of the NPF and NRT transporter families involved in nitrate uptake and transport have been identified and analyzed in maritime pine4. Among the transporter families, the NRT3 one is expanded and composed by six members. The capacity of maritime pine to use nitrate or ammonium has been analyzed in seedlings. The development and growth responses to nitrate nutrition are comparable to ammonium supply. At molecular level, there are strong gene expressions for genes involved in nitrate uptake and assimilation such as Nitrate Reductase, Nitrite Reductase, Glutamine Synthetase 1a, three NRT3 genes and different NPF family members in the different organs. Since the NPF proteins can transport different metabolites, peptides and hormones, the NPF transporters involved in nitrate transport are being identified.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. This project was supported by the grant MicroNUpE, BIO2015-73512-JIN; MINECO/AEI/FEDER, UE. JMVM was supported by a grant from the Spanish Ministerio de Educación y Formación Profesional (FPU17/03517) and FO by a grant from the Universidad de Málaga (Programa Operativo de Empleo Juvenil vía SNJG, UMAJI11, FEDER, FSE, Junta de Andalucía)

Correction to: Northerly wind trends along the Portuguese marine coast since 1950

All figure captions are not accurate. Also, there is a repeated error in most figures because the vertical scales incorrectly show Wind Strenght instead of Wind Strength and in Fig. 4 (mid-panel) 2003 instead of 2004.info:eu-repo/semantics/publishedVersio

An unexpected actor in ammonium assimilation in conifer trees

Conifers are tree species with enormous environmental and economic interests but with several characteristics that complicate their investigation (big size, secondary compounds, large long-life cycles, megagenomes…). However, they are well adapted to ammonium-rich soils being a good model to study ammonium assimilation in plants. Although they have a special feature, only two glutamine synthetase (GS, EC 6.3.1.2) genes, GS1a and GS1b, coding for cytosolic proteins, have been identified. In angiosperms and in the gymnosperm Ginkgo biloba there are two types of this enzyme responsible of the ammonium assimilation: GS1 expressed in the cytosol and GS2 in the plastids. Until the date, the searches of new GS1 and GS2 genes in conifers have been made with classical biochemical and molecular biology techniques without satisfactory results. In the present context, the emergence of the next generation sequencing (NGS) techniques has open new opportunities in the resolution of old problems. They have allowed the whole sequencing of the massive conifer genomes and the analysis of their transcriptomes. Thus, in the framework of the European project ProCoGen, a gene expression atlas of the tissues of one-month seedlings was carried out using laser capture microdissection (LCM) and massive sequencing in maritime pine (Pinus pinaster), which is a conifer tree from the Southwestern Mediterranean region1. From the analysis of this work, a new gene coding for a new putative cytosolic GS has been identified, PpGS1c. 1Cañas, RA et al. (2017). Plant J, 91. 1064-1087Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Project funding by Ministerio de Economía y Competitividad BIO2015-69285-R and MicroNUpE (BIO2015-73512-JIN; MINECO/AEI/FEDER, UE

Biotechnological approaches to increase biomass production in trees

Nutrient use efficiency is one of the factors influencing growth and therefore of high importance for biomass production in trees. Poplar is a model tree widely used for molecular and functional studies and the characterization of transgenic poplars overexpressing structural and regulatory genes involved in glutamine biosynthesis has provided insights on how glutamine metabolism is involved in N economy and biomass production in woody plant models. Numerous studies have shown the relevance of GS isoenzymes in plant development, biomass production, and yield (Cánovas et al. 2006; Castro-Rodríguez et al. 2015). In this communication two examples of functional analysis of plant genes in poplar, and their potential interest for biotechnological approaches are presented (Pascual et al. 2018; Rueda-López et al. 2017). Overexpression of cytosolic NADP+-isocitrate dehydrogenase (ICDH), one of the major enzymes involved in the production of 2-oxoglutarate for amino acid biosynthesis in plants, yields poplar trees with increased growth and enhanced vascular development in young leaves and apical stems. These plants also show an increased expression of genes associated with vascular differentiation and altered amino acids and organic acids content (Pascual et al. 2018). In other study, we observed that overexpression of Dof5, a transcriptional regulator of lignin production and the carbon-nitrogen balance, produced poplar trees with increased growth and biomass production when N availability in the soil is sufficient (Rueda-López et al. 2017). Taken together, these results suggest a close relationship between carbon and nitrogen metabolism and highlights the relevance of glutamine and glutamate biosynthesis in the control of growth and development. Research supported by Spanish Ministry of Economy and Competitiveness and Junta de Andalucía (Grants BIO2015-69285-R, BIO2012-0474 and research group BIO-114).Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Transcriptomics of ammonium nutrition in the conifer Pinus pinaster Aiton

Nitrogen is an important element for all living beings because it is part of macromolecules as significant as nucleic acids or amino acids. For plants, it constitutes a limiting factor in their growth and development1 due to their low natural availability in soils thus limiting primary production in ecosystems2. Conifers are a group of gymnosperm plants that form large forest extensions of vegetation, being the main constituents of forests in boreal ecosystems3 where ammonium is the main source of inorganic nitrogen4. Due to the characteristics of the soils in which conifers usually grow, these plants have developed a high tolerance to the presence of ammonium, which may constitute their main source of inorganic nitrogen5. The maritime pine (Pinus pinaster Aiton) is a conifer that has a wide distribution in the western Mediterranean area and has been widely used in reforestation, soil stabilization tasks and industrially. In recent years, maritime pine has been the subject of multiple omic studies that have resulted in the acquisition of important tools and resources6,7. The present work is focused on the analysis of the ammonium uptake and management efficiency, and its relationship with the biomass accumulation in maritime pine. For this purpose, several experiments have been developed in which pine seedlings have undergone different levels of ammonium nutrition, both in the short and long term. As a result of short-term experiments, the characterization of transcriptomic response to the process of ammonium nutrition (uptake and assimilation) is being studied at mRNA, lncRNA and miRNA level in roots. In relation to long-term experiments, ten different provenances of maritime pine seedlings were treated with different ammonium levels and the biomass changes were measured. The results obtained suggest the existence a certain phenotypic plasticity grade for this conifer.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. This project was supported by a grant form the Spanish Ministerio de Ciencia, Innovación y Universidades (MicroNUpE, BIO2015-73512-JIN; MINECO/AEI/FEDER, UE). FO was supported by a grant from the Universidad de Málaga (Programa Operativo de Empleo Juvenil vía SNJG, UMAJI11, FEDER, FSE, Junta de Andalucía) and JMVM by a grant from the Spanish Ministerio de Educación y Formación Profesional (FPU17/03517

Transcript profiling for early stages during embryo development in Scots pine

Background: Characterization of the expression and function of genes regulating embryo development in conifers is interesting from an evolutionary point of view. However, our knowledge about the regulation of embryo development in conifers is limited. During early embryo development in Pinus species the proembyo goes through a cleavage process, named cleavage polyembryony, giving rise to four embryos. One of these embryos develops to a dominant embryo, which will develop further into a mature, cotyledonary embryo, while the other embryos, the subordinate embryos, are degraded. The main goal of this study has been to identify processes that might be important for regulating the cleavage process and for the development of a dominant embryo. Results: RNA samples from embryos and megagametophytes at four early developmental stages during seed development in Pinus sylvestris were subjected to high-throughput sequencing. A total of 6.6 million raw reads was generated, resulting in 121,938 transcripts, out of which 36.106 contained ORFs. 18,638 transcripts were differentially expressed (DETs) in embryos and megagametophytes. GO enrichment analysis of transcripts up-regulated in embryos showed enrichment for different cellular processes, while those up-regulated in megagametophytes were enriched for accumulation of storage material and responses to stress. The highest number of DETs was detected during the initiation of the cleavage process. Transcripts related to embryogenic competence, cell wall modifications, cell division pattern, axis specification and response to hormones and stress were highly abundant and differentially expressed during early embryo development. The abundance of representative DETs was confirmed by qRT-PCR analyses. Conclusion: Based on the processes identified in the GO enrichment analyses and the expression of the selected transcripts we suggest that (i) processes related to embryogenic competence and cell wall loosening are involved in activating the cleavage process; (ii) apical-basal polarization is strictly regulated in dominant embryos but not in the subordinate embryos; (iii) the transition from the morphogenic phase to the maturation phase is not completed in subordinate embryos. This is the first genome-wide transcript expression profiling of the earliest stages during embryo development in a Pinus species. Our results can serve as a framework for future studies to reveal the functions of identified genes

Impact analysis of wind turbines blockage on doppler weather radar

Postprint (published version