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Objectives :

There is a great need to accelerate breeding in forest trees because of their long generation times and the projected demand for increased production of fibre. A clear understanding of the factors controlling wood formation and determining wood properties will also have major implications for the socio-economics of forest productivity. Wood properties are known to vary between species and among genotypes within species. This variability is heritable and thus presents an opportunity to select for superior wood properties and, as a result, for product quality. Such selection is currently hampered by costly traditional chemical and technological assays and the necessity to wait until the trees are nearly mature to evaluate wood properties. The combination of rapid methods for the chemical/physical characterisation of lignocellulosic fibre, pulp and paper characteristics, and genomic sciences to find the genes controlling these traits, should increase the efficiency of selection by shortening time and reducing costs associated with measuring such properties.

This project is specifically directed by market requirements of the pulp and paper industry and to a lesser extent the wood panel industry. It focuses on a single tree species: Maritime pine (Pinus pinaster Ait.) used as a model softwood, but results are expected to have broader applications for other conifers of major interest for the EU (Scots pine, Norway spruce). The major goal is to provide new strategic options for forest tree breeding by obtaining phenotypic and/or molecular selection criteria for genetic improvement of lignocellulosic fibre quality and, as a result, for product (pulp and paper) quality. This project will lead to the selection of elite trees for improved varieties deployment. The 9 organisations in this project come from the pulp, paper and biotechnology industries, research institutes and universities in five EU countries. Between them they have all the experience needed to complete the following objectives:

With respect to the first objective, the following tasks will be completed:

  1. Develop fast and cheap predictive tests to assay lignocellulosic fibre, pulp and paper characteristicsValidate these predictors againts measurements performed at the industrial level
  2. Determine the minimum amount of material (discs, cores, chips) required for a reliable predictive assay
  3. Test the within-tree variation for the predictive measurements
  4. Investigate the relationships between fine ultrastructural characteristics, pulp and paper properties
  5. Obtain estimates of genetic parameters (within and between provenance variability, heritability, genetic correlation between traits, parent-offspring correlations, genotype x environment interaction) of lignocellulosic fibre and pulping properties
  6. Select elite trees with high pulping quality

With respect to the second objective, genomic technologies will be developed to identify genes and proteins that control major component of wood quality. If we firmly believe that the mechanisms that determine wood properties and the information needed for practical application in trees must come from studies of trees, we also think that a deeper understanding of the molecular mechanisms involved in cell wall biosynthesis and assembly will arise from studies of the model plant system Arabidopsis thaliana (a small cruciferous weed). Therefore, genomic studies of both Maritime pine and Arabidopsis will be combined. The consortium will focus on achieving the following tasks:

  1. Sample differentiating Maritime pine xylem from contrasting wood types (early, late, juvenile, mature, side, compression wood)

  2. Construct composite normalized cDNA libraries from these tissues and array 10,000 plasmid clones in 96 well microtiter plates

  3. Provide 10,000 expressed sequence tags (EST) of Maritime pine xylem

  4. Construct high density filters (HDF) with non-redundant Maritime pine EST

  5. Study gene expression on HDF with mRNA extracted from xylem associated with diverse type of wood

  6. Identify and sequence differentially expressed genes/proteins from the contrasted xylem tissues

  7. Identify and sequence genes/proteins specifically expressed in xylem

  8. Identify "expressional" candidate genes (CG) involved in variation in wood properties from the sequence and expression data

  9. Provide access to and information on clones, libraries, sequences and expression data via an on-line relational database

  10. Screen novel cellulose mutants by FTIR in a collection of 20,000 Arabidopsis mutants

  11. Identify novel genes from cellulose mutants already known in Arabidopsis

  12. Identify Arabidopsis mutants for "expressional" (step 7) and "positional" (step 16) CG, and study their potential defects in the cell wall by analytical methods

  13. Develop a regeneration system via somatic embryogenesis and an efficient genetic transformation procedure in Maritime pine

  14. Transform Maritime pine embryonic lines with the most relevant CG identified in step 7 and verified in steps 11 and 16

  15. Analyze mutant Maritime pine trees by analytical methods

  16. Localise CG (defined in steps 7 and 10) on an existing Maritime pine genetic linkage map

  17. Study the co-locations between mapped CG and quantitative trait loci (QTL) for wood properties (QTLs detected in the framework of an ongoing EU FAIR project)

  18. Validate the CG of steps 11 and 16, by linkage disequilibrium study in natural populations of Maritime pine

  19. Select elite trees for improved pulp and paper properties using phenotypic and molecular diagnostic tools

We are aware of some of the limitations of the "positional" CG approach (steps 15 and 16) since some wood property QTLs may not segregate in the studied pedigree. In addition, it may not be possible to map some CG due to low levels or absence of sequence polymorphism. However, great genetic diversity is expected in a non-domesticated allogamous species like Maritime pine.