CBN-V Video Archives - S7-11
Cryopreservation of FEC Lines as an Aid for Genetic Transformation Programs

Escobar R.H.¹, L.G. Santos¹, J. Tohme¹ and WM. Roca²

1. Biotechnology Research Unit, International Center for Tropical Agriculture (CIAT), A.A. 6713, Cali, Colombia R.ESCOBAR@CGIAR.ORG
2. CIP, Apartado 1558, Lima, Peru

        Friable embryogenic callus lines were established at CIAT for some commercial Latin American cassava cultivars. In vitro-maintained FECs are genetically unstable, besides, keeping them is labor intensive and costly. Cryopreservation of FEC may provide a means of ensuring genetic stability of cell lines, and could be a source of “fresh” tissue useful for genetic transformation programs. To establish a cryopreservation protocol we are using cv. MCol 2215 and maintaining TMS 60444 as a control. Different cryopreservation methods were tested. Vitrification procedures did not work probably due to toxic effect of the cryo-protectan solution PVS2. Different temperatures of loading, PVS2 dilutions and pretreatment of FEC were tested, although tissue didn’t resume growth after freezing. Desiccation of FEC before freezing seems to have good potential to recover tissue. FEC grown on media with high agar content and growth regulators, desiccated during different times (10-30 days) are showing promising results after freezing. Frozen tissue was recovered on Greshoff and Doy solid medium supplemented with 50 M picloram, grown under low light intensity and transferred to liquid medium.

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For further information, see spoken presentation Escobar et al. S7-07

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