Keywords
liquid nitrogen
vitrification
in vitro culture
growth regulators
shoots
seeds
cider
cedrela odorata
crioconservación
nitrógeno líquido
vitrificación
cultivo in vitro
reguladores de crecimiento
ápices
semillas
cedro
cedrela odorata
How to Cite
Abstract
In order to protect the forest resources of Cedrela odorata L., currently threatened, it was considered necessary to establish a protocol for its cryopreservation. Research was conducted using shoots and seeds. Vitrification techniques were evaluated with shoots, using various solutions and rapid freezing in liquid nitrogen (LN). Initially the best results were obtained using incubation for 5 min in PVS3 solution before freezing, survival being 61% after 3 weeks of culture on Murashige and Skoog (MS) medium with 1.0 mg.l-1 BAP and 0.1 mg.l-1 GA3. Moreover, by incubating the apices in 0.3 M sucrose solution for 5 min prior to freezing, the survival rate reached 86% after 2 weeks of culture on MS medium with 1.0 mg.l-1 BAP, 0.1 mg.l-1 GA3 and 0.01 mg.l-1 IAA. When incubation in the solution of 0.3 M sucrose was used and recovery MS medium with 1 mg.l-1 KIN, 0.5 mg.l-1 GA3 and 100 mg.l-1 activated charcoal, the appearance of the surviving shoots improved significantly. For cryopreservation of cedar seeds, the methodology of dehydration and rapid freezing in liquid nitrogen (LN) was evaluated. Seeds with moisture content between 7.9% and 5.5% were placed in 5 ml polypropylene vials and these were directly immersed in NL. The effect of the storage period in NL (1 h, 24 h, 7 days, 1 month, 3 months and 6 months) on seed germination was evaluated. There were no statistical differences in ability of seeds to germinate after storage during different periods.References
ABDELNOUR A., AGUILAR M., VALVERDE L. 2011. Micropropagación de pilón (Hieronyma alchornoides). Agronomía Costarricense 32:9-19.
ABDELNOUR A., ROJAS G., ALFARO U. 2007. Estudios preliminares para la crioconservación de especies forestales arbóreas. Tecnología en Marcha 20:1-6.
AFANADOR A.M. 2005. Propagación in vitro a partir de meristemos de cinco variedades comerciales de Dianthus caryophyllus L., (clavel). Tesis de grado, Pontificia Universidad Javeriana. Bogota, Colombia. 54 p.
BENSON E., JOHNSTON J., MUTHUSAMY J., HARDING K. 2006. Physycal and engineering perspectives of in vitro plant cryopreservation. Plant Tissue Culture Engineering 5:441-476.
CARDOSO F., PITA J.M., PALMEIRA J. 2000. Efecto de la crioconservación sobre la germinación de semillas de leguminosas. Campiña Grande. Revista Brasileira de Produtos Agroindustriais 2:67-71.
COLLADO R., BARBÓN R., AGRAMONTE D., JIMÉNEZ F., PÉREZ M., GUTIÉRREZ O., RAMÍREZ D. 2004. Establecimiento in vitro de ápices y segmento nodales de Swietenia macrophylla King. Biotecnología Vegetal 4:143-146.
DÍAZ A., BRENES C., CASCANTE D., OVARES C. 2010. Plagas y enfermedades forestales en cedro amargo (Cedrela odorata). Escuela de Ciencias Ambientales. Universidad Nacional de Costa Rica, pp. 3, 8, 13.
DOLCE N., SCOCCHI A., MROGINSKI L. 2004. Crioconservación de ápices de Citrus sinensis. IBONE, Argentina. FACENA 20:37-41.
ENGELMANN F. 2000. Importance of cryopreservation for the conservation of plant genetic resources, pp. 8-20. In: F. Engelmann and H. Takagi (eds.). Cryopreservation of Tropical Plant Germplasm. Current research progress and application. IPGRI. Rome, Italy.
ENGELMANN F. 2011. Use of biotechnologies for the conservation of plant biodiversity. In Vitro Cellular and Developmental Biology Plant 47:5–16.
FLORES E. 1999. La Planta: estructura y función, pp. 663- 735. Vol. II. Libro Universitario Regional. Editorial Tecnológica de Costa Rica. Cartago, Costa Rica.
GAMBOA J.P., ABDELNOUR A. 1999. Micropropagación de melina (Gmelina arborea ROXB). Agronomía Costarricense 23:69-76.
GARCÍA L., DE FERIA M., ACOSTA K. 2007. Aspectos básicos de la conservación in vitro de germoplasma vegetal. Cuba. Biotecnología Vegetal 7:67-79.
GEORGE E., HALL M.A., DE KLERK G. 2008. Plant Propagation by Tissue Culture. Vol. 1. 3 ed. Springer. pp. 2-3.
GONZALEZ M.T., ENGELMANN F. 2006. Cryopreservation of plant germplasm using the encapsulationdehydration technique: review and case study on sugarcane. Cryo-Letters 27:155-168.
HOWARD F., MERIDA M. 2004. Hypsipyla grandella (Zeller). Universidad de Florida. Consultado el 24 de enero del 2010. Disponible en http://entnem.ifas.ufl.edu/creatures/trees/moths/mahogany_borer-spanish.htm
INTERNATIONAL SEED TESTING ASSOCIATION (ISTA). 2008. International Ruler For Seed Testing: The germination test. Seed Science and Technology. 7 p.
IRIONDO J.M. 2001. Conservación de germoplasma de especies raras y amenazadas (Revisión). Universidad Politécnica de Madrid. Invest. Agr.: Prod. Prot. Veg. 16:5-24.
MARTÍNEZ R., AZPIROZ H., RODRÍGUEZ J., CETINA M., GUTIÉRREZ M. 2003. Aplicación de la biotecnología en los recursos genéticos forestales. Revista Chapingo 9:3-5.
MURASHIGE T., SKOOG F. 1962. A revised medium for rapid growth and bioassays with tobacco tissues culture. Physiol Plant 15:473-497.
NAVARRO C. 2002. Genetic resources of Cedrela odorata L., and their efficient use in Mesoamerica. Universidad de Helsinki. Helsinki, Finlandia, p. 7.
PÉREZ J., MESÉN F., HILJE L., AGUILAR M.E. 2001. Desarrollo de un método de micropropagación aplicable a genotipos selectos de Cedrela odorata L. Optimización de la fase de multiplicación. CATIE. Turrialba, Costa Rica. Revista Forestal Centroamericana 38:67-71.
POEHLMAN J. 2005. Mejoramiento genético de las cosechas. Limusa. México, p. 257.
REYES X., VERA J. 2004. Efecto de reguladores de crecimiento en las diferentes fases de la propagación in vitro de la malanga (Xanthosoma sagittifolium L. Schott). UTEQ. Los Ríos, Ecuador. 10 p.
RIVERA A.L., VALBUENA B., HIDALGO R., MORENO J.D. 2008. Crioconservación de yemas de microtubérculos de papa Solanum tuberosum ssp., andigena mediante desecado de tejidos. Colombia. Revista Corpoica - Ciencia y Tecnología Agropecuária 9:37-38.
SAKAI A., ENGELMANN F. 2007. Vitrification, Encapsulation-Vitrification and Droplet-Vitrification: A Review. Cryo-letters 28:151-172.
SÁNCHEZ N., JIMÉNEZ V.M. 2010. Técnicas de conservación in vitro para el establecimiento de bancos de germoplasma en cultivos tropicales. Universidad de Costa Rica. Revista Agronomía Mesoamericana 21:196-198.
TAKAGI H. 2000. Recent developments in cryopreservation of shoot apices of tropical species, pp. 178-191. In: F. Engelmann and H. Takagi (eds.). Cryopreservation of Tropical Germplasm Current Research Progress and Application. IPGRI. Rome, Italy.
WADSWORTH F. 2000. Producción Forestal para América Tropical. Departamento de Agricultura de los Estados Unidos (USDA). Washington, DC, US. 628 p.
WANG Z., DENG X. 2004. Cryopreservation of shoot tips of citrus using vitrification: effect of reduced form of glutathione. Cryo-Letters 25:51-58.
WU Y., ENGELMANN F., ZHAO Y. 2000. Cryopreservation of apple in vitro germplasm in China, p. 474. In: F. Engelmann and H. Takagi (eds.). Cryopreservation of Tropical Germplasm Current Research Progress and Application. IPGRI, Rome, Italy.