Somatic Embryogenesis of Brazilian Conifer Podocarpus lambertii Klotzsch ex Endl.
P. lambertii is a Brazilian conifer indigenous to one of the 25 biodiversity hotspots of the world, the subtropical moist forest ecoregion of the Atlantic Forest Biome. Its conservation status is classified as near threatened, being desirable the development and use of biotechnological tools, such as somatic embryogenesis (SE). SE is a complex, multifactorial model system of molecular, biochemical, physiological and developmental steps of plant embryogenesis, and comprises a high performance in vitro regenerative system for both the mass clonal propagation of an elite genotype and/or the mass propagation of endangered plant germplasm. Protocols for plant regeneration based on SE have been established for many conifers species, but species from the closely related families Araucariaceae and Podocarpaceae are in general considered recalcitrant to SE. Mostly of the SE protocols for conifers have been developed for members of family Pinaceae. In this context, here we describe the first SE protocol for a species from family Podocarpaceae, the P. lambertii. This protocol is based on induction with immature zygotic embryos, followed by establishment and multiplication of proembryogenic masses on plant growth regulator-free culture medium, and maturation phases I and II. Procedures including explant preparation, PEM proliferation, and somatic embryo development are described in detail.
This work was supported by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina (FAPESC 14,848/2011-2, and 2780/2012-4). We also thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico for the scholarships provided for LNV (CNPq 150,513/2017-8) and MPG (CNPq 306,126/2013-3).
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