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

Jean-François Trontin (1) , Sandrine Debille (1) , Francis Canlet (1) , Luc Harvengt (1) , Marie-Anne Lelu-Walter (2) , Philippe Label (2) , Caroline Teyssier (2) , Marie-Claude Lesage Descauses (2) , Claire Le Mette (2) , Célia Miguel (3, 4, 5) , José de Vega-Bartol (3, 4, 5) , Mariagrazia Tonelli (3, 4, 5) , Raissa Santos (3, 4, 5) , Andrea Rupps (6) , Seyedeh Batool Hassani (6) , Kurt Zoglauer (6) , Elena Carneros (7) , Carmen Díaz-Sala (7) , Dolores Abarca (7) , Isabel Arrillaga (8) , Isabel Mendoza-Poudereux (8) , Juan Segura (8) , Concepción Avila (9) , Marina Rueda (9) , Javier Canales (9) , Francisco M. Cánovas (9)

1 Biotechnology and advanced Forestry Department, Genetics & Biotechnology group
2 AGPF - Unité de recherche Amélioration, Génétique et Physiologie Forestières
3 Forest Biotechnology Laboratory
4 Forest Biotechnology Laboratory
5 Instituto de Technologia Quimica e Biologica, Forest Biotechnology Laboratory
6 Institute of Biology
7 Department of Plant Biology
8 Dpto. Biología Vegetal, Facultad de Farmacia
9 Molecular Biology Laboratory

Reverse genetics, defined as ectopic candidate gene expression or silencing, has become an indispensable tool 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/).

Mots clés: maritime pine; somatic embryogenesis; transgenesis; reverse genetics; functional gene validation

Domaines: Biologie végétale; Biotechnologies

Jean-François Trontin, Sandrine Debille, Francis Canlet, Luc Harvengt, Marie-Anne Lelu-Walter, et al.. Somatic embryogenesis as an effective regeneration support for reverse genetics in maritime pine: the Sustainpine collaborative project as a case study. 2. International Conference of the IUFRO Working Party 2.09.02, "Somatic Embryogenesis and Other Vegetative Propagation Technologies", International Union of Forest Research Organisations (IUFRO). Vienne, AUT., Jun 2012, Brno, Czech Republic. ⟨hal-01268055⟩