Specific Objectives. 1) Germplasm characterization for haplotypic variation in genes of forestry interest in E. grandis, E. globulus and E.



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Development of an integrated genotypification platform for bioprospection of interesting candidates genes in MERCOSUR Eucalyptus germplasm Argentina (IB-IRB-EEA-Concordia-INTA) Brasil (Unidad Recursos Genéticos y Biotecnología-EMBRAPA) Paraguay (Facultad de Ciencias Agrarias, Centro Multidisciplinario de Investigaciones Tecnológicas / Universidad Nacional de Asunción) y Desarrollos Madereros de Paraguay SA Uruguay (Mundial Forestación SA) Biotech-sur

General Objective Establishment of a regional network for genomic application and association mapping of Eucalyptus forests to explore genetic base for wood development with industrial and energetic purposes

Specific Objectives 1) Germplasm characterization for haplotypic variation in genes of forestry interest in E. grandis, E. globulus and E. urophylla 2) Localization of genome-wide candidate genes and SNPs in the genetic maps 3) Development and validation of NIR calibration curves for prediction of wood properties (lignin content, S/G, extractives, pulping yield) 3) Development of clonal populations of selected clones for future association genetic studies 4) Development of pilot association genetic studies

Expected Results Genotyping system based on SNP determination in candidate genes (bead arrays) (300-500) and genome wide for genetic mapping/ pilot association mapping and germplasm characterization (high-troughput) Genetic maps including more SSR markers and SNPs Co-localization of SNPs (candidate genes/others) and QTL Identification of associated genes with phenotypic properties useful for energy production and Eucalyptus wood industry

Expected Results Data base with annotated candidates genes, QTLs and SNPs map position. Scientific information for MAS application in breeding programmes based on polymorphisms of these genes Clonal field trials of Eucalyptus grandis and E. globulus for future association genetic analysis NIR test models for wood properties prediction in different Eucalyptus species (E. grandis y E. globulus) Training

Association Mapping pilot Clonal population Development for future Association mapping QTLs Co-localization Candidates Genes SNPs genome wide Genotyping SNPs Candidate genes Genome wide Association populations E. globulus E. grandis Phenotyping NIR prediction calibration curves Segregating populations E. grandis, E. grandisxurophylla

Materials and Methods Genotyping: Eucalyptus NCBI data base/ Genolyptus: Data mining for SNPs Mapping Populations: Segregating populations 2 E. grandis, 2 E. grandis x E. urophylla Association population 1 clonal E. grandis 2 E. globulus High-throughput analysis: Oligonucletides bead arrays (locus specific/allele specific): 1) SNPs Selection (sample) 2) Screening of Populations: segregating and association Phenotyping: lignin content, S/G, extractives, pulping yield Prediction NIR curves: NIR Screenig of individuals, selection for chemical analysis, curves developments for E. globulus/grandis Mapping analysis: pseudo test cross for linkage mapping and QTL analysis

Activities Data Mining for SNPs discovery in Eucalyptus NCBI data base/ Genolyptus. Primer design for Illumina system (locus specific/allele specific) Polymorphic SSR selection between parents and evaluation in segregating populations (complete developing maps (2 E. grandis, 2 E. grandis x E. urophylla) Selection of SNPs in a representative germplasm sample through bead arrays Development of clonal association population for E. grandis, E. globulus: Development of NIR calibration curves for E. globulus and E. grandis: NIR Screenig of individuals, selection of divergent for chemical analysis (lignin content, S/G, extractives, pulping yield) Screening of Populations: segregating and association Linkage Mapping analysis (pseudo test cross) QTL mapping and pilot association mapping

Twinning Given the hypothesis that wood quality genetic determinants are shared between different forest tree species, this work will be of interest to other tree researchers and forest companies. In particular, those genes involved in lignin content and composition could be of interest to paper and construction/furniture industry as well as cellulose-based bioenergy industry Possible colaborations in: - forest tree genomics - genetic determinants of lignocellullose biosynthesis and degradation

Partners Argentina: INTA: IB, IRB, EEA Concordia-: Susana Marcucci Poltri Esteban Hopp Martín García, Cintia Acuña Pablo Pathauer Celina Braccini Juan DiezLucas Landi Santiago Becerro Eduardo Pablo Cappa Susana Torales Noga Zelener Martín Marcó Javier Obershelp Leonel Harrand Nicolás Alanis Mario Carmarán Brasil EMBRAPA: Unidad Recursos Genéticos y Biotecnología: Darío Grattapaglia, Darío Grattapaglia, Danielle Alves de Faria, Georgios Pappas, Carolina Sansaloni, Cesar Daniel Petroli Paraguay Facultad de Ciencias Agrarias, Centro Multidisciplinario de Investigaciones Tecnológicas / Universidad Nacional de Asunción Mirtha Vera de Ortiz, Elizabeth Monges, Manuel M. Enciso G., María Liz García, Nancy Villalba, César Cardozo, Inocencia Peralta, Héctor Nakayama, Rafael Gómez Desarrollos Madereros de Paraguay SA José Elizaul, Guillermo Salvatierra, Ricardo Kiriluk, Valeria Escalada Uruguay Mundial Forestación SA: Nuno Borralho,Javier Burgueño, Fabio Abreu, Carmen Sobrera, Sebastian Gonzalez, Analia Vazques,