Vitamins C and E improve regrowth and reduce lipid peroxidation of blackberry shoot tips following cryopreservation
Vitamins C and E improve regrowth and reduce lipid peroxidation of blackberry shoot tips following cryopreservation
复制标题
DOI：
10.1007/s00299-009-0795-y
复制
发表时间：
2009
影响因子：
6.2
通讯作者：
E. Uchendu;S. Leonard;M. Traber;B. Reed
中科院分区：
文献类型：
--
作者：
E. Uchendu;S. Leonard;M. Traber;B. Reed
冷冻保存方案中涉及的氧化过程可能是导致液氮暴露后组织活力降低的原因。对抗这些反应的抗氧化剂应该可以改善恢复。本研究的重点是氧化脂质损伤和外源维生素E（生育酚，维生素E）和维生素C（抗坏血酸，维生素C）处理再生的植物玻璃化溶液2号（PVS 2）玻璃化冷冻保存技术的四个关键步骤的影响;预处理，加载，冲洗，和再生。初步实验表明，当在四个步骤中的任何一个步骤添加时，11-15 mM的维生素E显着增加再生（P< 0.001）。有显着更多的丙二醛（MDA），脂质过氧化产物，在每个步骤比新鲜的未处理的茎尖。结果表明，处理后的芽中α-和γ-生育酚含量显著高于未处理的芽。在每个步骤添加维生素E显着降低MDA的形成和提高芽再生。维生素C（0.14-0.58 mM）也显着改善再生芽尖在每个步骤相比，控制。高铁浓度和维生素C的再生培养基降低回收率。然而，在无铁培养基中，维生素C显着提高回收率。用维生素E（11 mM）和维生素C（0.14 mM）组合的处理并不显著优于单独的维生素C。我们建议在PVS 2玻璃化方案中向预处理培养基、上样液或冲洗液中添加维生素C（0.28 mM）。这是第一次报告的应用维生素，以改善低温保存的植物组织，最大限度地减少氧化损伤。
Oxidative processes involved in cryopreservation protocols may be responsible for the reduced viability of tissues after liquid nitrogen exposure. Antioxidants that counteract these reactions should improve recovery. This study focused on oxidative lipid injury and the effects of exogenous vitamin E (tocopherol, Vit E) and vitamin C (ascorbic acid, Vit C) treatments on regrowth at four critical steps of the plant vitrification solution number 2 (PVS2) vitrification cryopreservation technique; pretreatment, loading, rinsing, and regrowth. Initial experiments showed that Vit E at 11–15 mM significantly increased regrowth (P< 0.001) when added at any of the four steps. There was significantly more malondialdehyde (MDA), a lipid peroxidation product, at each of the steps than in fresh untreated shoot tips. Vit E uptake was assayed at each step and showed significantly more α- and γ-tocopherols in treated shoots than those without Vit E. Vit E added at each step significantly reduced MDA formation and improved shoot regrowth. Vit C (0.14–0.58 mM) also significantly improved regrowth of shoot tips at each step compared to the controls. Regrowth medium with high iron concentrations and Vit C decreased recovery. However, in iron-free medium, Vit C significantly improved recovery. Treatments with Vit E (11 mM) and Vit C (0.14 mM) combined were not significantly better than Vit C alone. We recommend adding Vit C (0.28 mM) to the pretreatment medium, the loading solution or the rinse solution in the PVS2 vitrification protocol. This is the first report of the application of vitamins for improving cryopreservation of plant tissues by minimizing oxidative damage.