Home / Actions supported / Projects / Cryopreservation of endangered American plants
AMCRYO

Cryopreservation of endangered American plants

Developing the plant of the future
Export to PDF

Abstract

This project aimed at developing cryopreservation protocols for in vitro shoot tips of vanilla and Clinopodium odorum using new, vitrification-based techniques. In case of Clinopodium, we applied the vitrification (V) cryo-plate protocol for cryopreservation of in vitro-derived shoot tips. We studied the recovery of shoot tips after modification of various parameters of the V cryo-plate protocol, including sucrose concentration in the preconditioning medium, composition of the loading solution (LS) and vitrification solution (VS), and duration of treatment with PVS2 VS, which contained (w/v) 30% glycerol, 15% dimethylsulfoxide, 15% ethylene glycol, and 13.7% sucrose. We also compared the efficiency of the V cryo-plate protocol with the dehydration (D) cryo-plate protocol. The optimal conditions determined for the V cryo-plate protocol included a 24-h preconditioning treatment on medium with 0.3 M sucrose; treatment for 20 min at room temperature with LS containing 2.0 M glycerol and 0.4 M sucrose; and dehydration with PVS2 for 60 min at 0°C. Under these conditions, 71.0% recovery of cryopreserved shoot tips was achieved. Only 29.2% regeneration was noted with the D cryo-plate protocol.
- As regards vanilla, we compared the droplet-vitrification technique and the V cryo-plate protocol. Recovery of a few cryopreserved apices could be achieved using the V cryo-plate protocol. However, due to the very slow growth of the apices, a sufficient number of replications could not be made to allow determining optimal experimental conditions.

Perspectives

Cryopreservation is ready tob e applied to Clinopodium shoot tips.

Project Number : 1202-007

Year : 2012

Type of funding : AAP OS

Project type : AAP

Start date :
01 Mar 2013

End date :
30 Sep 2014

Flagship project :
Non

Project leader :
Florent Engelmann

Project leader's institution :
IRD

Project leader's RU :
DIADE

Budget allocated :
50000 €

Total budget allocated ( including co-financing) :
50000 €

Funding :
Labex