Long-term preservation of Lotus tenuis adventitious buds
Fecha
2019Autor
Espasandin, Fabiana Daniela
Brugnoli, Elsa Andrea
Ayala, Paula Gabriela
Ayala, Lilian P.
Ruiz, Oscar Adolfo
Sansberro, Pedro Alfonso
Metadatos
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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.
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