Ascorbic acid addition during dehydration improves garlic shoot tip cryopreservation but does not affect viral load
Ascorbic acid addition during dehydration improves garlic shoot tip cryopreservation but does not affect viral load
复制标题
脱水过程中添加抗坏血酸可改善大蒜茎尖冷冻保存，但不影响病毒载量
DOI：
10.1016/j.cryobiol.2022.05.001
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Zhen Wu
中科院分区：
文献类型：
--
作者：
Xiaodong Xing;Min Liu;Rong Zhou;Fangling Jiang;Yunhe Bai;Hanyu Wei;Deng Zhang;Jingjing Wei;Zhen Wu
超低温保存是大蒜脱毒的有效方法。然而，超低温保存过程中的氧化损伤严重影响大蒜超低温保存后的存活。抗坏血酸（阿萨）可以减少低温保存后的氧化损伤并促进再生，其效果可能受到添加步骤的影响。在本研究中，阿萨在冷冻保存的脱水保护（O）和脱水（DE）步骤中加入。通过观察不同浓度的阿萨处理对茎尖细胞活力和活性氧（ROS）组分的动态变化，证实了阿萨与茎尖ROS积累减少有关。抗氧化酶基因表达水平的增加也解释了阿萨清除ROS的作用。细胞活力、ROS、膜脂过氧化相关指标和抗氧化剂相关指标之间的相关性分析表明，过量ROS引起的膜脂过氧化是影响细胞活力的主要因素。在脱水过程中加入抗坏血酸可以减少从脱水到稀释过程中活性氧的积累，减轻低温保存过程中的氧化损伤。因此，大蒜超低温保存后的存活率和再生能力得到显著提高。脱水是大蒜超低温保存过程中添加阿萨的适宜步骤。我们进一步评估了在最佳阿萨处理下的病毒清除效果。但再生植株中病毒含量与对照相比无显著差异。脱水是大蒜超低温保存中添加阿萨的最佳步骤。·抗坏血酸促进茎尖清除活性氧，影响抗氧化酶编码基因的表达。·在脱水步骤中添加阿萨显著改善了冷冻保存后的存活和再生长。
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. • Dehydration is the optimal step for adding AsA in garlic cryopreservation. • Ascorbic acid promotes the ROS scavenging of shoot tip and influences the expression of antioxidant enzyme encoding genes. • The addition of AsA to the dehydration step significantly improves the survival and regrowth after cryopreservation.