Preliminary study on cryopreservation of Dendrobium Bobby Messina protocorm- like bodies by vitrification

• Main Authors: James Antony, Jessica Jeyanthi, Chan, Lai Keng, Rathinam, Xavier, Sinniah, Uma Rani, Subramaniam, Sreeramanan
• Format: Article
• Language: English
• Published: Academic Journals 2010
• Online Access: http://psasir.upm.edu.my/id/eprint/11372/1/Preliminary_Study_of_Cryopreservation_of_Dendrobium_Bobby_Messina_Protocorm-like_Bodies_by_Vitrification.pdf; http://psasir.upm.edu.my/id/eprint/11372/; http://www.academicjournals.org/journal/AJB/article-abstract/358261823850
• Institution: Universiti Putra Malaysia

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.