Abstract
<em>Ceratopteris thalictroides</em> 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.
Highlights
Cryopreservation is currently the best method for longterm, ex situ conservation of plant genetic resources and involves the use of liquid nitrogen (LN; −196°C)
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
Our present study focused on a fern species which is sensitive to both low temperature and dehydration
Summary
Cryopreservation is currently the best method for longterm, ex situ conservation of plant genetic resources and involves the use of liquid nitrogen (LN; −196°C). 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 present paper describes a simple method for the successful cryopreservation of this highly sensitive to low temperature and dehydration stress fern species.
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