Culture treatments for enhancing post-thaw recovery of cryopreserved suspension cells of potato cv. Desiree.
An efficient and reproducible protocol has been developed for the cryopreservation of cell suspension cultures of the potato (Solanum tuberosum L.) cv. Desiree. An evaluation was made of the effectiveness of different pre-culture and post-thaw treatments on cell growth, as measured by changes in biomass. Cell suspensions were cultured in UM medium supplemented with 0.25, 0.5, 0.625, 0.75 or 1.0 M sucrose prior to cryopreservation. Sucrose-treated cells were harvested from suspension and 0.75 ml packed cell volumes placed in 2 ml capacity polypropylene vials with 0.5 ml of chilled cryoprotectant (glycerol 46.0 g 1(-1), dimethylsulphoxide 39.0 g 1(-1), sucrose 342.0 g 1(-1) proline 5.0 g 1(-1); pH 5.8). Cells were frozen at -0.5 degrees C min(-1) from 0 to -35 degrees C, held at -35 degrees C for 35 min and stored, for 10 days, in liquid nitrogen (-196 degrees C). The most effective pre-treatment, in terms of subsequent post-thaw cell viability as assessed by fluorescein diacetate uptake or triphenyltetrazolium chloride reduction, was culture with 0.75 M sucrose. For this treatment, the mean absorbance (490 nm) following triphenyltetrazolium chloride reduction was 88% greater (p < 0.05) than control and 59% greater (p < 0.05) for thawed cells also cultured on supporting filter paper discs.
Sadia
Bushra
B
Plant Sciences Division, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough LE12 5RD, UK.
Anthony
Paul
P
Lowe
Kenneth C
KC
Power
J Brian
JB
Davey
Michael R
MR
D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000737:chemistry*; D004121:Dimethyl Sulfoxide / Q000737:chemistry*; D005452:Fluoresceins / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry*; D011198:Solanum tuberosum / Q000166:cytology* / Q000378:metabolism; D013395:Sucrose / Q000737:chemistry; D013778:Tetrazolium Salts / Q000737:chemistry
D003451:Cryoprotective Agents; D005452:Fluoresceins; D013778:Tetrazolium Salts; D013395:Sucrose; C009591:triphenyltetrazolium; D005990:Glycerol; C018506:diacetylfluorescein; D004121:Dimethyl Sulfoxide