Articles
Adventitious bud clusters can increase tissue cryopreservation efficiency – a case study of gesneriads
Article number
1421_11
Pages
77 – 86
Language
English
Abstract
Exceptional species cannot be conserved in seed banks and will require cryopreservation for their long-term ex situ conservation.
For some, in vitro tissues will be needed, particularly for many exceptional species that are not producing seeds or are desiccation sensitive.
It is not known how many species will require this approach, but estimates suggest they will number in the thousands, and efficient methods are needed to reduce the time and costs for accomplishing their conservation.
One of the most time-consuming steps in cryopreserving plant tissues is the isolation of shoot tips to cryopreserve shoot meristems.
With some species, however, it is possible to generate multiple buds that can be easily prepared for cryopreservation protocols.
Two examples come from the Gesneriaceae: species of Cyrtandra and of Saintpaulia. Multiple adventitious buds were generated on leaves taken from in vitro-grown cultures of C. gracilis, C. kaulantha, S. brevipilosa, S. confusa, S. grandifolia, S. rupicola, and S. tongwensis by culture on MS medium with 2 mg L‑1 IAA and 0.1 mg L‑1 BAP. Small clusters of bud tissue were isolated and successfully cryopreserved using droplet vitrification, in the case of Cyrtandra, and encapsulation dehydration, in the case of the Saintpaulia species.
Survival through cryopreservation ranged from 0 to 50% in Cyrtandra and from 38 to 80% with Saintpaulia species, depending on the experiment and the species.
These results demonstrate the feasibility of using such bud clusters as targets for cryopreservation to reduce the time for tissue excision.
Similar efficiencies can be obtained using regenerative callus or fern green globular bodies, but the ability to generate any of these tissues more broadly is currently limited.
However, this approach may warrant further research, to reduce the cost and time required for cryopreservation protocols, to facilitate the conservation of the many threatened exceptional species that will require cryobanking for their long-term ex situ conservation.
For some, in vitro tissues will be needed, particularly for many exceptional species that are not producing seeds or are desiccation sensitive.
It is not known how many species will require this approach, but estimates suggest they will number in the thousands, and efficient methods are needed to reduce the time and costs for accomplishing their conservation.
One of the most time-consuming steps in cryopreserving plant tissues is the isolation of shoot tips to cryopreserve shoot meristems.
With some species, however, it is possible to generate multiple buds that can be easily prepared for cryopreservation protocols.
Two examples come from the Gesneriaceae: species of Cyrtandra and of Saintpaulia. Multiple adventitious buds were generated on leaves taken from in vitro-grown cultures of C. gracilis, C. kaulantha, S. brevipilosa, S. confusa, S. grandifolia, S. rupicola, and S. tongwensis by culture on MS medium with 2 mg L‑1 IAA and 0.1 mg L‑1 BAP. Small clusters of bud tissue were isolated and successfully cryopreserved using droplet vitrification, in the case of Cyrtandra, and encapsulation dehydration, in the case of the Saintpaulia species.
Survival through cryopreservation ranged from 0 to 50% in Cyrtandra and from 38 to 80% with Saintpaulia species, depending on the experiment and the species.
These results demonstrate the feasibility of using such bud clusters as targets for cryopreservation to reduce the time for tissue excision.
Similar efficiencies can be obtained using regenerative callus or fern green globular bodies, but the ability to generate any of these tissues more broadly is currently limited.
However, this approach may warrant further research, to reduce the cost and time required for cryopreservation protocols, to facilitate the conservation of the many threatened exceptional species that will require cryobanking for their long-term ex situ conservation.
Authors
V.C. Pence, M. Philpott
Keywords
Cyrtandra, exceptional plant, ex situ conservation, in vitro, Saintpaulia
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