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A Simple Microaspiration Technique for Isolating Somatic Cells from Cryopreserved Equine Semen as Nuclear Donors for Cloning
Published on: December 19, 2025
Simplified and improved somatic embryogenesis for clonal propagation of Pinus pinaster (Ait.)
Marie-Anne Lelu-Walter1, Michele Bernier-Cardou, Krystyna Klimaszewska
1Unité Amélioration, Génétique et Physiologie Forestières, INRA-CRO BP 20 619, Ardon F-45166, Olivet Cedex, France. Leluwa@orleans.inra.fr
Abstract:
In this study, several improvements and simplifications of SE protocols in Pinus pinaster (Ait.), a species of economic importance in the regions of Western Europe, are described. These improvements pertained to all stages of SE including high initiation frequencies in eight control pollinated seed families, relatively high somatic embryo maturation yield when cells were coated with particles of activated charcoal and a rapid production of plants directly in a shade house. The SE initiation frequency from isolated zygotic embryos was high (up to 100%) and plants were produced from 11 embryogenic lines representing all crosses. Based on these results, the estimated number of somatic embryos required to produce 1,000 plants varied from slightly more than the required number of plants to more than double this number depending on the line. Such an estimate is critical in developing plant production strategy when a number of embryogenic lines are considered for production of clonal plants.
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