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Simplified and improved somatic embryogenesis of hybrid larches (Larix × eurolepis and Larix × marschlinsii). Perspectives for breeding

Simplification et amélioration de l’embryogenèse somatique des mélèzes hybrides (Larix × eurolepis et Larix × marschlinsii). Perspectives pour l’amélioration

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Abstract

  • • Development of clonal propagation method, such as somatic embryogenesis, has numerous applications such as mass-production of genetically improved plants and the amenability of embryogenic cultures to cryogenic storage. Since the 90’s, researchers at INRA have engaged in research on somatic embryogenesis in Larix species (Larix × eurolepis, Larix × marschlinsii).

  • • The aim of this work was to improve and to simplify all steps of somatic embryogenesis and to apply this protocol to the new hybrid variety REVE-VERT.

  • • The somatic embryogenesis initiation frequency from immature zygotic embryos was high (65%) on a medium with reduced plant growth regulator concentrations (2.2 μM of 2.4-dichlorophenoxyacetic acid and 2.3 μM of 6-benzyladenine). Simplified cryopreservation method (no need of programmable freezer) of the embryonal masses resulted in 100% recovery of cryopreserved lines. Maturation of a large number of somatic embryos was greatly improved when embryonal masses were dispersed on filter paper placed on medium containing high concentration of gellan gum (8 g·L−1). Under these conditions, 94% of the lines matured somatic embryos that developed into plantlets. Clearly ageing and cryopreservation did not reduce embryogenic potential of embryonal masses.

  • • Requirements for the effective integration of somatic embryogenesis into the larch breeding programme are discussed.

Résumé

  • • Le développement de méthode de multiplication clonale, telle l’embryogenèse somatique, a de nombreuses applications comme la production en masse de plants génétiquement améliorés et la disponibilité des cultures embryogènes en cryoconservation. Depuis les années 1990, l’INRA a engagé des recherches en embryogenèse somatique des mélèzes (Larix × eurolepis, Larix × marschlinsii).

  • • L’objectif du travail a été de simplifier et d’améliorer chaque étape de l’embryogenèse somatique, et d’appliquer ce protocole à la nouvelle variété de mélèze hybride REVE-VERT.

  • • L’initiation de l’embryogenèse somatique à partir d’embryons zygotiques immatures est obtenue à des fréquences élevées (65 %) avec un milieu de culture contenant des concentrations hormonales réduites (2,2 μM d’acide 2.4-dichlorophenoxyacetic et 2,3 μM de 6-benzyladenine). Une méthode simplifiée de congélation (sans l’utilisation d’un congélateur programmable) des masses embryogènes permet la survie de 100 % des lignées congelées. La maturation d’un grand nombre d’embryons somatiques a été fortement améliorée lorsque les masses embryogènes ont été étalées sur filtre papier placé sur milieu de culture à forte concentration en gelrite (8 g·L−1). Sous ces conditions, 94 % des lignées embryogènes régénèrent des embryons somatiques matures qui se développent en plantes. Clairement l’âge et la cryoconservation des lignées embryogènes ne réduisent pas leur potentiel embryogène.

  • • Les besoins pour une intégration efficace de l’embryogenèse somatique au programme d’amélioration du mélèze, sont discutés.

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Authors and Affiliations

  1. UR 588 Research Unit on Breeding, Genetic and Physiology of Forest Trees, INRA, 2163 Avenue de la Pomme de pin CS 4001, Ardon, F-45075, Orléans Cedex 2, France

    Marie-Anne Lelu-Walter & Luc E. Pâques

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  1. Marie-Anne Lelu-Walter
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  2. Luc E. Pâques
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Correspondence to Marie-Anne Lelu-Walter.

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Lelu-Walter, MA., Pâques, L.E. Simplified and improved somatic embryogenesis of hybrid larches (Larix × eurolepis and Larix × marschlinsii). Perspectives for breeding. Ann. For. Sci. 66, 104 (2009). https://doi.org/10.1051/forest/2008079

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  • DOI: https://doi.org/10.1051/forest/2008079

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