A simple way to overcome the recalcitrance of the water fern Ceratopteris thalictroides (L.) Brongn. to cryopreservation

• Main Authors: Damian Makowski, Jan J. Rybczyński, Anna Mikuła
• Format: Article
• Language: English
• Published: Polish Botanical Society 2015-10-01
• Series: Acta Societatis Botanicorum Poloniae
• Subjects: abscisic acid; alginate bead size; encapsulation-dehydration; darkness; pre-culture; regrowth
• Online Access: https://pbsociety.org.pl/journals/index.php/asbp/article/view/4803

Ceratopteris thalictroides is a water fern very sensitive to both dehydration and low temperature. This study focuses on the cryopreservation of this species by encapsulation-dehydration technique, in particular on the effects of pre-culture step, alginate bead size and the physical conditions of culture on the cryopreservation efficiency. Encapsulated and non-pre-cultured gametophytes did not survive cooling with liquid nitrogen. When cryopreservation was preceded by a 2-week period of pre-culture, regrowth reached 42.1%. Reduction in the size of the alginate bead, and culture in total darkness resulted in improved gametophyte regrowth capacity (75.5% or 81.7%, respectively). The best results (91.3%) were obtained when all factors tested occurred simultaneously. The gametophytes recovered very quickly and sporophytes were formed within 4 weeks after rewarming. These simple improvements can be used, not only for the cryopreservation of gametophytes in cryptogams but also for some recalcitrant species of seed plants.