Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration

Shoot-tips from several genotypes of the cardenolide-producing perennial shrub Digitalis obscura L. were successfully cryopreserved using the encapsulation-dehydration technique. Precultivation on MS medium containing 0.5 M sucrose, followed by 2.5 h dehydration (final weight 30%) induced shoot regr...

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Autores principales: Sales, Ester, Nebauer, Sergio G., Arrillaga, I., Segura, J.
Formato: article
Lenguaje:Inglés
Publicado: 2017
Acceso en línea:http://hdl.handle.net/20.500.11939/4469
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author Sales, Ester
Nebauer, Sergio G.
Arrillaga, I.
Segura, J.
author_browse Arrillaga, I.
Nebauer, Sergio G.
Sales, Ester
Segura, J.
author_facet Sales, Ester
Nebauer, Sergio G.
Arrillaga, I.
Segura, J.
author_sort Sales, Ester
collection ReDivia
description Shoot-tips from several genotypes of the cardenolide-producing perennial shrub Digitalis obscura L. were successfully cryopreserved using the encapsulation-dehydration technique. Precultivation on MS medium containing 0.5 M sucrose, followed by 2.5 h dehydration (final weight 30%) induced shoot regrowth in 42% of cryopreserved shoot-tips. Cold-hardening of the in vitro cultures before sucrose treatment dramatically increased shoot recovery up to 86%. The optimized cryopreservation protocol was then employed using different shoot cultures from five D. obscura genotypes. Responses to cryopreservation depended mainly on the genotype, best results being obtained when shoot tips from HU3 and LL11 were used. Prolonged subcultures reduced proliferation rates in both control and cryopreserved HU3 shoot-tips, whereas long-term storage in LN did not affect the shoot recovery rate of the genotype. RAPID markers were employed to evaluate possible somaclonal variation occurring in shoots regenerated through successive subcultures and after cryo preservation. The band patterns revealed differences between the original parent plant and the shoots grown in vitro, especially after a prolonged subculture (84.9% of matches for HU3 after 16 subcultures vs. 93.4% for AY3 after 2 subcultures). Nevertheless, match percentages were higher (98.6% to 99.5%) when band patterns from subcultured shoots were compared to those obtained from their respective control or frozen progenies indicating that cryopreservation ensure genetic stability of selected Digitalis obscura genotypes.
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spelling ReDivia44692025-04-25T14:43:37Z Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration Sales, Ester Nebauer, Sergio G. Arrillaga, I. Segura, J. Shoot-tips from several genotypes of the cardenolide-producing perennial shrub Digitalis obscura L. were successfully cryopreserved using the encapsulation-dehydration technique. Precultivation on MS medium containing 0.5 M sucrose, followed by 2.5 h dehydration (final weight 30%) induced shoot regrowth in 42% of cryopreserved shoot-tips. Cold-hardening of the in vitro cultures before sucrose treatment dramatically increased shoot recovery up to 86%. The optimized cryopreservation protocol was then employed using different shoot cultures from five D. obscura genotypes. Responses to cryopreservation depended mainly on the genotype, best results being obtained when shoot tips from HU3 and LL11 were used. Prolonged subcultures reduced proliferation rates in both control and cryopreserved HU3 shoot-tips, whereas long-term storage in LN did not affect the shoot recovery rate of the genotype. RAPID markers were employed to evaluate possible somaclonal variation occurring in shoots regenerated through successive subcultures and after cryo preservation. The band patterns revealed differences between the original parent plant and the shoots grown in vitro, especially after a prolonged subculture (84.9% of matches for HU3 after 16 subcultures vs. 93.4% for AY3 after 2 subcultures). Nevertheless, match percentages were higher (98.6% to 99.5%) when band patterns from subcultured shoots were compared to those obtained from their respective control or frozen progenies indicating that cryopreservation ensure genetic stability of selected Digitalis obscura genotypes. 2017-06-01T10:10:12Z 2017-06-01T10:10:12Z 2001 DEC 2001 article Sales, E., Nebauer, S. G., Arrillaga, I., Segura, J. (2001). Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration. Planta Medica, 67(9), 833-838. 0032-0943 http://hdl.handle.net/20.500.11939/4469 10.1055/s-2001-18861 en openAccess Impreso
spellingShingle Sales, Ester
Nebauer, Sergio G.
Arrillaga, I.
Segura, J.
Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration
title Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration
title_full Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration
title_fullStr Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration
title_full_unstemmed Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration
title_short Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration
title_sort cryopreservation of digitalis obscura selected genotypes by encapsulation dehydration
url http://hdl.handle.net/20.500.11939/4469
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