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The MicroRNAs in Eucalyptus tension wood world
Perspectives of extension of this common project to a more ambitious proposal to be submitted to funding agencies : Long term Added value of the cooperation : The project will benefit from the collaboration of internationally renowned laboratories. It is very important for their complementary technical and scientific expertise, the availability of certified clones and installation of the experimental device. This project will also greatly benefit from technical and scientific expertise and synergies resulting from the collaboration between the international partners IICT and UMR5546 in the areas of phenotype characterization, functional genomics, bioinformatics, molecular biology and forest genetics. Advanced training will be also promoted in form of PhD and Master thesis. The project was able to attract the support of an international consultant with renowned expertise on the field of Eucalyptus genomics and miRNA regulation.
Summary of the proposal :
Wood is a complex and highly variable tissue, which anatomical, chemical, physical and technological features are ontogenic and environmentally controlled. In response to a gravitropic stimulus, Angiosperms differentiate a tension wood. This type of wood shows specific characteristics such as the presence of a gelatinous layer (layer-G), with high cellulose content (~95%) presenting high crystallinity, and their microfibrils are oriented nearly parallel to axis of the fibers. Tension wood is considered an excellent model to study the regulation of the formation of xylem cell walls (eg biosynthesis and deposition of cellulose, lignin and other biopolymers) and how this regulation is reflected in the final properties of wood. The ability to drive wood quality is fundamental for Portuguese economy. Portugal is one of the largest producers of pulp derived from eucalyptus (E. globulus), with exports accounting for about 5% of GDP. It is also relevant that E. globulus represents about 1/3 of the Portuguese forest and that the use of adequate genotypes will optimize the exploitation of available areas. The understanding of the molecular mechanisms involved in the biosynthesis of the cell wall is of great importance not only for future production of pulp and paper, but also for the production of bio-fuels and bio-materials. Many of the genes involved in wood formation have been catalogued, but the mechanisms to regulate this process of development are still far from being elucidated. The discovery of microRNAs (miRNAs) and their functions opened new frontiers for a better knowledge on the mechanisms of regulation of many developmental processes, including cambium differentiation. However, of the approximately 8,600 miRNA available in the database miRbase (http://microrna.sanger.ac.uk/sequences/), none of the entries relate to the most economically relevant Portuguese forest species, such as Pinus pinaster, Quercus suber and Eucalyptus globulus. This 3 year project combines the phenotypic characterization of the developing xylem with the latest genomic strategies/tools available, such as next generation sequencing technologies, gene expression analysis, bioinformatics, physical mapping, and functional analysis. In this project, we will identify and characterize E. globulus miRNAs involved in the regulation mechanisms of wood formation and their target genes, using as a model the tension wood forming tissues. The long term aim is to use this information to devise new ways to control the quality of wood produced by E. globulus and to provide the breeding programs with tools to direct their work to the selection/production of genotypes with desired wood qualities. The specific objectives of this proposal include: 1) Development and application of new non-destructive technologies and methodology to chemical characterize wood forming tissues; 2) anatomical characterization of wood forming tissues subject to a gravitropic stimulus; 3) Production of expressed sequences catalogues, in particular miRNAs catalogues, and to characterize de genomic sequences of these transcripts; 4) validation of miRNA candidates and their target; 5) To provide spatial expression information for selected transcription factors and miRNA; 6) to map target genes and miRNA onto physical maps; 7) to provide new insights trough functional analysis in transient and stable transgenic system; 8) To boost the competitiveness of national institutions in national and international scientific and technical panorama, by reinforcing and establishing new national and international collaborations with industry and academia.
Antonio PINEDA-LUCENA, Valencia Claudio GOMES, Lisboa
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