LONG-TERM PRESERVATION OF LOTUS TENUIS ADVENTITIOUS BUDS

ESPASANDIN FD; BRUGNOLI E.A.; AYALA P.G.; AYALA P.G.; RUIZ O.A.; SANSBERRO P

PLANT CELL TISSUE AND ORGAN CULTURE

SPRINGER

Lugar: Berlin; Año: 2019 vol. 136 p. 373 - 382

0167-6857

Encapsulation-dehydration, encapsulation-vitrification, and vitrification were tested for cryopreservation of Lotus tenuis (Fabaceae) adventitious buds clusters (ABCs) obtained by a direct regeneration from leaves cultures. Among them, only 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 30min. 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 ten 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-cryopreserved control. Thus, our results indicate that the use of vitrification-based PVS3 solution offers a simple, accurate, and appropriate procedure for the cryopreservation of Lotus tenuis adventitious buds.