info:eu-repo/semantics/article
Long-term preservation of Lotus tenuis adventitious buds
Fecha
2019-02Registro en:
Espasandin, Fabiana Daniela; Brugnoli, Elsa Andrea; Ayala, Paula Gabriela; Ayala, Lilian Patricia Elizabeth; Ruiz, Oscar Adolfo; et al.; Long-term preservation of Lotus tenuis adventitious buds; Springer; Plant Cell, Tissue and Organ Culture; 136; 2; 2-2019; 373-382
0167-6857
1573-5044
CONICET Digital
CONICET
Autor
Espasandin, Fabiana Daniela
Brugnoli, Elsa Andrea
Ayala, Paula Gabriela
Ayala, Lilian Patricia Elizabeth
Ruiz, Oscar Adolfo
Sansberro, Pedro Alfonso
Resumen
Encapsulation-dehydration, encapsulation-vitrification, and vitrification were tested for cryopreservation of Lotus tenuis (Fagaceae) adventitious buds clusters (ABCs) obtained by a direct regeneration system from leaves cultures. Among them, the PVS3-based vitrification procedure was found to be useful for survival and regrowth of the preserved explants. For vitrification, the ABCs were dehydrated in a solution containing 2 M glycerol + 0.4 M sucrose for 25 min at room temperature, submerged in PVS3 solution for 1 h at 0 °C, then immersed in liquid nitrogen for 48 h and rapidly rewarmed. Afterword, the explants were unloaded in MS liquid medium with 1.2 M sucrose for 30 min. The washed tissues were dried superficially on filter paper and cultured in semisolid hormone-free MS medium containing 0.1 M sucrose. All cultures were maintained at 25 °C in the dark for 10 days and transferred to the light conditions. With this procedure, 79 ± 5.3% survival and more than 80% of the plantlets displaying a phenotype similar to the non-treated control after acclimatization. The data settled from ISSR showed no genetic dissimilarities between in vitro regenerants derived from cryopreserved tissues and the non-treated plants. Thus, our results indicate that the use of vitrification-based PVS3 solution offers a simple, accurate, and appropriate procedure for the cryopreservation of L. tenuis adventitious buds.