Regulación coordinada de los genes implicados en la síntesis de fenilalanina en pino

Regulación coordinada de los genes implicados en la síntesis de Fenilalanina en pino Craven-Bartle B, Pascual MB, Cánovas FM, Avila C Departamento de Biología Molecular y Bioquímica, Facultad de Ciencias, Campus Universitario de Teatinos, Universidad de Málaga, 29071-Málaga, España ([email protected]) Durante el ciclo vital de coníferas como el pino marítimo (Pinus pinaster Ait.) una gran cantidad de esqueletos carbonados se ven inmovilizados de forma irreversible en la madera. Este es un proceso muy costoso en términos energéticos en el que el carbono de la fotosíntesis se canaliza a través de la vía del Siquimato para la biosíntesis de los fenilpropanoides. Esta ruta metabólica fundamental está finamente regulada principalmente a través de control de la transcripción, y puesto que la fenilalanina es el precursor para la biosíntesis de los fenilpropanoides, la regulación precisa de la síntesis de fenilalanina y su utilización debe ocurrir simultáneamente. Los tres promotores de los genes que codifican las enzimas, Prefenato Aminotransferasa (PAT), Fenilalanina Amonio Liasa (PAL), y la Glutamina Sintetasa (GS1b), contienen elementos AC que participan en la activación transcripcional mediada por factores de R2R3-Myb. En este trabajo hemos examinado la capacidad de los factores de transcripción R2R3-Myb: Myb1, Myb4 y Myb8 para co-regular la expresión de PAT, PAL y GS1b. Sólo Myb8 es capaz de activar la transcripción de los tres genes. Por otra parte, la expresión de este factor de transcripción es mayor en tejidos lignificados, donde hay una gran demanda de fenilpropanoides. En un experimento de ganancia de función, hemos demostrado que Myb8 puede unirse específicamente un elemento bien conservado tipo AC-II, de ocho nucleótidos de longitud en las regiones promotoras de PAT, PAL y GS1b, activando de ese modo su expresión. Nuestros resultados muestran que Myb8 regula la expresión de estos genes implicados en el metabolismo de la fenilalanina, que se requiere para la canalización de carbono fotosintético para promover la formación de la madera. La co-localización de los tránscritos de PAT, PAL, GS1b y MYB8 en células vasculares también apoya esta conclusión. Financiado por: Proyecto de excelencia de la Junta de Andalucía (CVI-3739), Proyecto del Ministerio de Ciencia e Innovación (BIO2009-07490) y por el programa KBBE Plant (proyecto SUSTAINPINE)Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Root and shoot growth of Pinus strobus x P. wallichiana somatic plants is influenced by the nitrogen composition of the germination medium

Somatic embryogenesis (SE) in conifers has proven to be an useful biotechnological tool for species conservation and mass propagation, but to make it commercially viable, adaptive research is required to convert laboratory protocols into plantation reality. Hybrid white pines are of interest to forest industry in North America due to the potential resistance to white pine blister rust (Cronartium ribicola). Somatic embryos of many pine species are typically germinated on the same nutrient medium as used for earlier stages of SE. For most of the genotypes of white and hybrid white pine, the time required for the plantlets to grow a 2 cm root takes approximately five to six months at which time the plantlets can be potted and transferred to a greenhouse. The survival of plants is however sketchy and not always reliable. In order to accelerate and improve the root growth and survival of the plants in a greenhouse a study was initiated on the manipulation of the nitrogen composition of the germination medium. We were interested in learning whether the pine somatic seedlings had a preference for organic or inorganic forms of N during the germination step and root growth. Experiments have been conducted to determine the uptake and content of 15N in those roots by submerging them in a hydroponic solution for two hours. The somatic seedlings’ root growth was dramatically affected by the N composition of the medium as well as it did affect the uptake of 15N. To follow up the nitrogen metabolism in somatic seedlings, we have performed microarray and qPCR analysis for nitrogen or nitrogen-related metabolism genes. The results of the analysis will be discussed.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Nitrogen metabolism in forest trees

NITROGEN METABOLISM IN FOREST TREES Francisco Cánovas, Concepción Ávila, Fernando N de la Torre, Rafael Cañas, Belén Pascual. Universidad de Málaga Email:[email protected] Forests are essential components of the ecosystems covering approximately one-third of the Earth’s land area and playing a fundamental role in the regulation of terrestrial carbon sinks. Forest trees are also of significant economic importance, as they are used for timber and paper production worldwide. A sustainable management of forest resources is needed to preserve natural forest and to meet the increasing international demands in the production of wood and the other forest-derived products. New advances and developments in biotechnology will contribute to accelerate the domestication of important traits for forest productivity. It is critical to identify the fundamental constraints on forest productivity to addressing these constraints with modern genomic tools. Nitrogen availability extremely low in forest ecosystems, and consequently, forest trees have evolved adaptive mechanism and biotic interactions to guarantee the strict economy of this essential nutrient. Nitrogen assimilation and recycling play a key role in the tree growth and biomass production and we firmly believe that knowledge on nitrogen metabolism will lead to approaches aimed at increasing forest productivity. In our laboratory, we are interested in studying nitrogen metabolism and its regulation the conifer maritime pine (Pinus pinaster Aiton), a forest tree species of great economic and ecological importance in the Mediterranean area and relevant model for conifer genomic research in Europe. Current research efforts are focused on improving the understanding of the response of conifer trees to ammonium availability and the transcriptional control of ammonium assimilation into amino acids. An overview and update 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, PLE2009-016 and research group BIO-114).Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Impactos ambientais do manejo agroecológico da caatinga no Rio Grande do Norte.

O objetivo deste trabalho foi avaliar os impactos ambientais do manejo agroecológico da caatinga, em unidades de produção familiar no Rio Grande do Norte, pelo método Ambitec de produção animal - dimensão ambiental, desenvolvido pela Embrapa Meio Ambiente. Foram avaliadas sete unidades de produção familiar, em quatro projetos de assentamentos de reforma agrária do Município de Apodi, RN. Os dados para o levantamento foram obtidos por meio de questionários aplicados aos representantes das unidades produtivas familiares, que atribuíram, a cada variável estudada, um valor que representou a alteração proporcionada pela implementação da tecnologia. Após a inserção dos coeficientes de alteração de cada variável dos indicadores por unidade de produção, o coeficiente de impacto foi automaticamente calculado por meio da planilha Ambitec. O manejo agroecológico da caatinga resultou num impacto ambiental positivo, e suas maiores contribuições foram relacionadas aos efeitos positivos dos seguintes indicadores: capacidade produtiva do solo, uso de insumos materiais, qualidade do produto e diminuição da emissão de poluentes à atmosfera. Dois indicadores geraram efeitos negativos: o uso de energia e o uso de recursos naturais. Pela superioridade dos benefícios gerados, o manejo agroecológico da caatinga é uma inovação tecnológica geradora de impactos ambientais positivos

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Involvement of a Myb transcription factor in the regulation of phenylalanine pathway in maritime pine

Wood is traditionally among the most important commercial products because of the high demand that exits for its derivatives. Trees, including conifers, divert large quantities of carbon into the biosynthesis of phenylpropanoids, particularly to generate lignin. Although lignin and other phenolic compounds do not contain nitrogen, phenylalanine metabolism is required to channel photosynthesis-derived carbon to phenylpropanoid biosynthesis. The phenylpropane skeleton required for lignin biosynthesis is provided by the deamination of phenylalanine in the reaction catalysed by the enzyme phenylalanine ammonia-lyase (PAL). This reaction is quantitatively important in trees because lignin biosynthesis is required for wood formation, and it releases large quantities of ammonium. An efficient and coordinated pathway for the amination of prephenate and the deamination of phenylalanine should be operative in lignifying cells to provide phenylalanine for lignin biosynthesis, and to re-assimilate ammonium. We hypothesized that one way to ensure efficient photosynthetic carbon channeling for lignin and other phenylpropanoid biosynthesis, together with nitrogen recycling, would be to couple both processes in time and in space by transcriptionally regulating the genes involved in phenylalanine biosynthesis and use. The experiments described in this communication attempt to test this hypothesis. To this end, we have isolated the promoter region of the three genes involved in the phenylalanine pathway in Pinus pinaster: PAL, GS1b and PAT. We have conducted both in vitro and in vivo studies using three different Myb transcription factors: PtMyb1, PtMyb4 from P. taeda and PpMyb8 from P. pinaster. We have studied the possible coupling in space and time of gene products for the operative co-regulation of both processes in pine trees, and have proven that Myb8 is a potential candidate to be the transcriptional regulator of phenylalanine metabolism in P. pinaster vascular cells.UNIVERSIDAD DE MÁLAGA. CAMPUS DE EXCELENCIA INTERNACIONAL ANDALUCÍA TEC