Preliminary study on cryopreservation of Dendrobium Bobby Messina protocorm- like bodies by vitrification
James Antony, Jessica Jeyanthi and Chan, Lai Keng and Rathinam, Xavier and Sinniah, Uma Rani and Subramaniam, Sreeramanan (2010) Preliminary study on cryopreservation of Dendrobium Bobby Messina protocorm- like bodies by vitrification. African Journal of Biotechnology, 9 (42). pp. 7063-7070. ISSN 1684–5315
Protocorm-like bodies of Dendrobium Bobby Messina were cryopreserved by vitrification method. In this study, protocorm-like bodies (PLBs) with the size range of 1-2 and 3-4 mm were selected from 4 weeks old culture, pretreated with half strength semi-solid Murashige and Skoog (MS) media supplemented with 0.5 M sucrose at 25°C for 24 h. Pretreated PLBs were then treated with a mixture of 2 M glycerol and 0.4 M sucrose supplemented with half strength liquid MS media at 25°C for 20 min. Osmoprotected PLBs were then dehydrated with plant vitrification solution 2 at 0°C for 20 min before storage in liquid nitrogen. After rapid warming in water bath at 40°C for 90 s, the PLBs were washed with half strength liquid MS media supplemented with 1.2 M sucrose and then cultured on half strength semi-solid MS media supplemented with 2% sucrose without the presence of any growth regulators. Survival of the cryopreserved PLBs was assessed based on triphenyl tetrazoliumchloride (TTC) spectrophotometrical analysis. The PLBs with 3-4 mm size range showed better viability comparative to size range 1-2 mm for both cryopreserved and non-cryopreserved PLBs. The best pretreatment concentration used in pretreatment media was 0.6 M sucrose and 1.2 M sorbitol, respectively.
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