《植物生理学报》 2015, 51(11): 1880-1886

外源抗氧化剂对春石斛类原球茎超低温保存的影响

狄玮, 贾梦雪, 刘燕^*

花卉种质创新与分子育种北京市重点实验室, 国家花卉工程技术研究中心, 城乡生态环境北京实验室, 北京林业大学园林学院, 北京100083

通信作者：刘；E-mail: chblyan@163.com；Tel: 010-62336062

摘　要：

为探讨外源抗氧化剂对春石斛类原球茎(PLBs)超低温保存中氧化应激的缓解效应, 以春石斛‘红梦幻’ PLBs为试材, 在玻璃化法超低温保存程序的不同环节导入3种不同浓度外源抗氧化剂, 检测其对PLBs冻后存活率的影响, 在此基础上分析最佳抗氧化剂介入实验体系中氧化应激相关指标的变化。结果显示, 在装载、PVS2处理和去装载时导入不同浓度的超氧化物歧化酶(SOD)和过氧化氢酶(CAT)对PLBs冻后存活率影响差异大, 均以去装载时导入效果最好。其中SOD导入最佳浓度为600 U•mL^-1, 存活率提高了5.76%, 但对氧化应激相关指标均无显著影响。CAT导入最佳浓度为400 U•mL^-1, 存活率提高了33.49%, 显著降低过氧化氢(H₂O₂)和丙二醛(MDA)含量, 提高抗坏血酸(AsA)含量。在预培养、装载、PVS2处理和去装载时导入不同浓度的AsA均可提高PLBs冻后存活率, 以去装载时导入效果最好, 最佳浓度为0.1 mmol•L^-1, 存活率提高了14.34%, 显著降低了超氧阴离子(O₂ •–)产生速率和MDA含量, 提高了SOD活性。结果表明, 3种外源抗氧化剂对PLBs冻后存活率相对提高效果依次为CAT>AsA>SOD。

关键词：春石斛; 类原球茎; 存活率; 抗氧化剂; 氧化应激

收稿：2015-06-19   修定：2015-09-06

资助：国家自然科学基金(31370693)。

Effects of Exogenous Antioxidants on Cryopreservation of Protocorm-Like Bodies of Dendrobium nobile

DI Wei, JIA Meng-Xue, LIU Yan^*

College of Landscape Architecture, Beijing Forestry University, Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, National Engineering Research Center for Floriculture, Beijing Laboratory of Urban and Rural Ecological Environment, Beijing 100083, China

Corresponding author: LIU Yan; E-mail: chblyan@163.com; Tel: 010-62336062

Abstract:

The aim was to explore the alleviating effects of oxidative stress in cryopreservation of protocorm-like bodies (PLBs) of Dendrobium nobile by importing exogenous antioxidants. PLBs of Dendrobium nobile ‘Hamana Lake Dream’ were used as materials for cryopreservation by vitrification, in which were added three kinds of different concentrations of exogenous antioxidants to investigate the effects on the survival rates, the best antioxidants-applied experiment systems were established respectively to research the changes of oxidative stress related indexes. The results showed that the impact on survival rates of cryopreservated PLBs were suffered a big difference by adding different concentrations of superoxide dismutase (SOD), catalase (CAT) to the loading, PVS2-treated and unloading stage. However, both for adding in the unloading stage were the best, In this stage, adding 600 U•mL^-1 SOD had the best effects, improved the survival rates of 5.76%, but all oxidative stress related indexes did not make a big difference. Adding 400 U•mL^-1 CAT had the best effects, improved the survival rates of 33.49%, the contents of hydrogen peroxide (H₂O₂) and malonaldehyde (MDA) decreased, and the contents of ascorbic acid (AsA) increased significantly. Adding different concentrations of AsA in the preculture, loading, PVS2-treated and unloading stages all could improve survival rates of cryopreservated PLBs. The best concentrations was 0.1 mmol•L^-1 in the unloading stage, improved the survival rates of 14.34%, the contents of O₂ •– (superoxide anion) production rate and MDA decreased significantly, and the activity of SOD increased significantly. The results indicated that the effects of three kinds of exogenous antioxidants on survival rates of cryopreservated PLBs were CAT>AsA>SOD in order.

Key words: Dendrobium nobile; PLBs; survival rates; antioxidants; oxidative stress