Effect of PVS2 vitrification on Brassidium shooting star orchid using protocorm-like bodies (PLBs)

• Main Authors: James Antony, Jessica Jeyanthi, Burkhan, Hazirah, Sinniah, Uma Rani, Poobathy, Ranjetta, Subramaniam, Sreeramanan
• Format: Article
• Language: English
• Published: Southern Cross Publishing 2013
• Online Access: http://psasir.upm.edu.my/id/eprint/29390/1/Effect%20of%20PVS2%20vitrification%20on%20Brassidium%20shooting%20star%20orchid%20using%20protocorm-like%20bodies%20%28PLBs%29.pdf; http://psasir.upm.edu.my/id/eprint/29390/; http://www.cropj.com/july2013.html

A cryopreservation procedure was developed to preserve protocorm-like bodies (PLBs) of Brassidium Shooting Star using the PVS2 vitrification technique. The optimised protocol involved the preculture of 3-4mm PLBs in half-strength Murashige and Skoog (MS) semi-solid medium supplemented with 0.8M sucrose, followed by dehydration in PVS2 solution for 20 minutes at 0°C, prior to storage in liquid nitrogen. The viability of non-cryopreserved and cryopreserved PLBs was determined by the 2,3,5-triphenyltetrazolium chloride (TTC) assay, after two weeks of recovery. The chlorophyll contents, total soluble protein and peroxidase activities of both non-cryopreserved and cryopreserved PLBs were assayed after three weeks of recovery. The results from the biochemical analyses indicated that control PLBs produced the highest viability, followed by treatment on non-cryopreserved PLBs (-LN) and cryostored PLBs (+LN), except in the peroxidase activity assay. The peroxidase activity was detected as the highest in cryostored PLBs followed by treated but non-cryopreserved PLBs, and control PLBs.