Xing Xiaodong, Liu Min, Zhou Rong, Jiang Fangling, Bai Yunhe, Wei Hanyu, Zhang Deng, Wei Jingjing, Wu Zhen
College of Horticulture, Nanjing Agricultural University, 1 Weigang, Nanjing, 210095, China; Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in East China, Ministry of Agriculture, Nanjing, China.
College of Horticulture, Nanjing Agricultural University, 1 Weigang, Nanjing, 210095, China; Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in East China, Ministry of Agriculture, Nanjing, China.
Cryobiology. 2022 Aug;107:64-73. doi: 10.1016/j.cryobiol.2022.05.001. Epub 2022 May 11.
Cryopreservation is known be an effective method for virus elimination in garlic. However, oxidative damage during the cryopreservation seriously affects the survival of garlic after cryopreservation. Ascorbic acid (AsA) can reduce oxidative damage and improve regrowth following cryopreservation, and its effect may be influenced by the step during which it is added. In this study, AsA was added at the osmoprotection (O) and dehydration (DE) steps of cryopreservation. By observing the dynamic changes in cell viability and reactive oxygen species (ROS) components with different AsA treatments, AsA has been linked to the reduced accumulation of ROS in the shoot tips. Increased gene expression levels of antioxidant enzymes also explained the ROS scavenging effect of AsA. The correlation analysis between cell viability, ROS, membrane lipid peroxidation-related indicators and antioxidant-related indicators showed that membrane lipid peroxidation caused by excess ROS was the main factor affecting cell viability. Ascorbic acid added during dehydration minimized the accumulation of ROS from dehydration to dilution and alleviated the oxidative damage during cryopreservation. Thus, the survival and regrowth of the garlic was significantly improved after cryopreservation. Dehydration was found to be the suitable step for the addition of AsA during garlic cryopreservation. We further evaluated the virus elimination effect under optimal AsA treatment. However, there was no significant difference in virus content in regenerated plants when compared with the control.
冷冻保存是一种有效的大蒜病毒消除方法。然而，冷冻保存过程中的氧化损伤严重影响了冷冻保存后大蒜的存活率。抗坏血酸（AsA）可以减少氧化损伤并提高冷冻保存后的再生能力，其效果可能受添加步骤的影响。在本研究中，在冷冻保存的渗透保护（O）和脱水（DE）步骤中添加AsA。通过观察不同AsA处理下细胞活力和活性氧（ROS）成分的动态变化，发现AsA与茎尖中ROS积累的减少有关。抗氧化酶基因表达水平的提高也解释了AsA的ROS清除作用。细胞活力、ROS、膜脂过氧化相关指标与抗氧化相关指标的相关性分析表明，过量ROS引起的膜脂过氧化是影响细胞活力的主要因素。在脱水过程中添加抗坏血酸可使从脱水到解冻过程中ROS的积累最小化，并减轻冷冻保存过程中的氧化损伤。因此，冷冻保存后大蒜的存活率和再生能力显著提高。发现脱水是大蒜冷冻保存过程中添加AsA的合适步骤。我们进一步评估了最佳AsA处理下的病毒消除效果。然而，与对照相比，再生植株中的病毒含量没有显著差异。
