Plant regeneration in Stone pine (<Emphasis Type="Italic">Pinus pinea</Emphasis> L.) by somatic embryogenesis |
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Authors: | E Carneros C Celestino K Klimaszewska Y-S Park M Toribio J M Bonga |
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Institution: | 1.Instituto Madrile?o de Investigación y Desarrollo Rural, Agrario y Alimentario (IMIDRA),Madrid,Spain;2.Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre,Québec,Canada;3.Natural Resources Canada, Canadian Forest Service, Canadian Wood Fibre Centre,Fredericton,Canada |
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Abstract: | Regeneration of plants by somatic embryogenesis (SE) was achieved in Stone pine (Pinus pinea), one of the most characteristic tree species of the Mediterranean ecosystem. The initial explants were megagametophytes
containing zygotic embryos from five selected half-sib families collected at different dates over 2 consecutive years. Rates
of extrusion and initiation of SE differed in both years. However, qualitative patterns were very similar: for most families,
the responsive developmental window was from late cleavage polyembryony to early cotyledonary stage. The highest overall mean
frequencies of extrusion and SE initiation (7 and 0.9%, respectively, for the five families and the eight 2006 collections)
were obtained on a modified Litvay’s medium with 9 μM 2,4-D and 4.5 μM BAP, supplemented with L-glutamine and casein hydrolysate. Families showed large differences in frequencies of SE initiation from year to year. Only
seven embryogenic lines were induced in 2005, representing three of the five families tested, whereas 34 lines from all the
families were obtained in 2006. Proliferation of embryonal masses (EM) was significantly improved when they were subcultured
after dispersing in liquid medium and collected on filter paper disks, instead of being subcultured as small clumps. This
effect showed a significant interaction with genotype. Several preconditioning treatments and culture media combinations were
tested for embryo development and maturation. The high proliferation rate of EM hampered somatic embryo development. However,
up to 42 mature embryos from different lines of three of the five families were obtained, 23 of them germinated and seven
converted into somatic seedlings. |
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Keywords: | Conifer Embryonal mass Genetic effect Half-sib families Somatic embryogenesis Somatic seedling Tree breeding |
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