Somatic embryogenesis as an effective regeneration support for reverse genetics in maritime pine: the Sustainpine collaborative project as a case study

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Trontin, Jean-François | Debille, Sandrine | Canlet, Francis | Harvengt, Luc | Lelu-Walter, Marie-Anne | Label, Philippe | Teyssier, Caroline | Lesage Descauses, Marie-Claude | Le Mette, Claire | Miguel, Célia | de Vega-Bartol, José | Tonelli, Mariagrazia | Santos, Raissa | Rupps, Andrea | Hassani, Seyedeh Batool | Zoglauer, Kurt | Carneros, Elena | Díaz-Sala, Carmen | Abarca, Dolores | Arrillaga, Isabel | Mendoza-Poudereux, Isabel | Segura, Juan | Avila, Concepción | Rueda, Marina | Canales, Javier | Cánovas, Francisco M.

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Reverse genetics, defined as ectopic candidate gene expression or silencing, has become an indispensabletool for functional dissection of traits of interest in forest trees. In maritime pine as in other conifers, long generation time and long life span, high genetic loads as well as high genetic redundancy are major obstacles to perform standard genetic approaches including association genetics. Validating marker associations with specific properties before transferring into breeding selection models is still challenging. Stable Agrobacterium-mediated genetic transformation of maritime pine was first reported by Trontin et al. (2002) and developed in both France (FCBA, INRA) and Portugal (IBET) with sufficient refinement (reviewed in Trontin et al. 2007) to envisage practical application in reverse genetics as an attractive alternative to association studies. As a result, the technology has been recently implemented in French (GenoQB, 2006-2009) and transnational (Sustainpine, 2010-2013) or European initiatives (ProCoGen 2012-2016). Somatic embryogenesis (SE) was revealed as a key tissue culture system for achieving genetic transformation, easy cryopreservation of transgenic tissue and efficient transgenic plant regeneration in maritime pine. Much consideration is given in France, Portugal and Spain to apply SE as a clonal propagation system for maritime pine improvement and deployment strategies in multivarietal forestry (MVF, Park YS, reviewed in Klimaszewska et al. 2007). By using this technology it is expected that we will obtain greater genetic gain, high flexibility in deployment of tested embryogenic varieties and easy balancing of genetic gain and diversity in plantations. Both SE and Agrobacterium-mediated transformation methods developed at FCBA, INRA or IBET are being successfully transferred to different partners through running the Sustainpine project (http://www.scbi.uma.es/sustainpine/).

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