School of Biological Sciences, Universiti Sains Malaysia (USM), 11800 Gelugor, Pulau Pinang, Malaysia.
Appl Biochem Biotechnol. 2013 Jul;170(5):1066-79. doi: 10.1007/s12010-013-0241-z. Epub 2013 May 3.
Dendrobium sonia-28 is an important ornamental orchid in the Malaysian flower industry. However, the genus faces both low germination rates and the risk of producing heterozygous progenies. Cryopreservation is currently the favoured long-term storage method for orchids with propagation problems. Vitrification, a frequently used cryopreservation technique, involves the application of pretreatments and cryoprotectants to protect and recover explants during and after storage in liquid nitrogen. However, cryopreservation may cause osmotic injuries and toxicity to cryopreserved explants from the use of highly concentrated additives, and cellular injuries from thawing, devitrification and ice formation. Reactive oxygen species (ROS), occurring during dehydration and cryopreservation, may also cause membrane damage. Plants possess efficient antioxidant systems such as the superoxide dismutase (SOD) and catalase (CAT) enzymes to scavenge ROS during low temperature stress. In this study, protocorm-like bodies (PLBs) of Dendrobium sonia-28 were assayed for the total protein content, and both SOD and CAT activities, at each stage of a vitrification exercise to observe for deleterious stages in the protocol. The results indicated that cryopreserved PLBs of Dendrobium sonia-28 underwent excessive post-thawing oxidative stress due to decreased levels of the CAT enzyme at the post-thawing recovery stage, which contributed to the poor survival rates of the cryopreserved PLBs.
铁皮石斛 sonia-28 是马来西亚花卉产业中一种重要的观赏兰花。然而,该属植物存在发芽率低和产生杂合后代的风险。目前,对于繁殖问题较多的兰花,冷冻保存是一种受欢迎的长期储存方法。玻璃化法是一种常用的冷冻保存技术,它涉及到使用预处理和冷冻保护剂来保护和恢复在液氮中储存期间和之后的外植体。然而,冷冻保存可能会导致渗透损伤和毒性,这是由于使用高浓度添加剂对冷冻保存的外植体造成的,还可能会因解冻、玻璃化和冰晶形成而导致细胞损伤。在脱水和冷冻保存过程中产生的活性氧(ROS)也可能导致膜损伤。植物具有有效的抗氧化系统,如超氧化物歧化酶(SOD)和过氧化氢酶(CAT)酶,可在低温胁迫下清除 ROS。在这项研究中,对铁皮石斛 sonia-28 的原球茎样体(PLBs)在玻璃化过程的每个阶段进行总蛋白含量和 SOD 和 CAT 活性的测定,以观察该方案中有害阶段。结果表明,由于解冻后恢复阶段 CAT 酶水平降低,冷冻保存的铁皮石斛 sonia-28 PLBs 在解冻后经历了过度的氧化应激,这导致冷冻保存的 PLBs 存活率较低。