17 January 2024 | Daniel Ballesteros, María Teresa Martínez, Carolina Sánchez-Romero, Itziar Aurora Montalbán, Ester Sales, Paloma Moncaleán, Isabel Arrillaga and Elena Corredoira
The article reviews the current status of cryopreservation techniques for embryogenic material of woody species, including conifers, fruit species, deciduous forest species, and palms. Cryopreservation, which involves storing cells or tissues at liquid nitrogen temperatures (-196°C), is crucial for long-term conservation of embryogenic material without loss of their capacity to regenerate. Despite significant advancements, large-scale conservation in cryobanks remains limited. Cryopreservation facilitates the management of embryogenic lines, reducing maintenance costs and the risk of somaclonal variation or contamination. Somatic embryogenesis, combined with cryopreservation, is particularly useful for preserving juvenility while field-testing corresponding clones. Traditional methods include slow cooling and vitrification, with desiccation-based procedures being simpler but less widely used. Genetic stability is supported by multiloci PCR-derived markers, though DNA methylation status assays show some changes after cryopreservation. The article details different cryopreservation procedures and their genetic stability after liquid nitrogen storage, highlighting the importance of selecting appropriate initial explants, preconditioning treatments, cryoprotection methods, and rewarming protocols. It also discusses the application of these techniques in various woody species, emphasizing the role of cryopreservation in conservation, breeding, and biotechnological manipulations.The article reviews the current status of cryopreservation techniques for embryogenic material of woody species, including conifers, fruit species, deciduous forest species, and palms. Cryopreservation, which involves storing cells or tissues at liquid nitrogen temperatures (-196°C), is crucial for long-term conservation of embryogenic material without loss of their capacity to regenerate. Despite significant advancements, large-scale conservation in cryobanks remains limited. Cryopreservation facilitates the management of embryogenic lines, reducing maintenance costs and the risk of somaclonal variation or contamination. Somatic embryogenesis, combined with cryopreservation, is particularly useful for preserving juvenility while field-testing corresponding clones. Traditional methods include slow cooling and vitrification, with desiccation-based procedures being simpler but less widely used. Genetic stability is supported by multiloci PCR-derived markers, though DNA methylation status assays show some changes after cryopreservation. The article details different cryopreservation procedures and their genetic stability after liquid nitrogen storage, highlighting the importance of selecting appropriate initial explants, preconditioning treatments, cryoprotection methods, and rewarming protocols. It also discusses the application of these techniques in various woody species, emphasizing the role of cryopreservation in conservation, breeding, and biotechnological manipulations.