论文标题
使用谷胱甘肽添加植物玻璃化溶液PVS+
Cryopreservation of seeds of blue waterlily (Nymphaea caerulea) using glutathione adding plant vitrification solution, PVS+
论文作者
论文摘要
Nymphaea caerulea是一种有价值的淡水水生植物，这不仅是因为其装饰价值，而且还因为其用于化学和医疗用途的提取。有必要将种子作为备用资源存储。冷冻保存是长期保存植物遗传库的可靠，具有成本效益的方法，尤其是对于顽固植物而言。在这项研究中，我们证明，由于无法忍受干燥和低温，牛乳杆菌是顽固的。由于在存储6个月之前丢失了生存力，因此通过传统存储方法不适合长期存储。长期储存N. caerulea种子的唯一方法是冷冻保存。然而，植物玻璃化溶液3（PVS3； 50％w/v蔗糖，50％w/v甘油）是一种常用的植物玻璃化溶液，对牛乳杆菌种子的冷冻保存无效。由PVS3处理的冷冻保存种子的最大存活率仅为23％。由活性氧（ROS）在种子中积累引起的氧化应激是导致PVS3处理的种子冷冻保存效率低下的原因。我们通过将谷胱甘肽（GSH）添加到PVS3中，开发了一种新的植物玻璃化溶液（PVS+）。 PVS+通过将ROS的积累和冷冻保存种子的存活率降低到97％，从而挽救了效率低下。我们的结果表明，植物组织冷冻保存效率低下可能是由ROS诱导的氧化应激引起的。抗氧化剂可以减少ROS诱导的氧化应激，并改善冷冻保存后的种子存活。总之，念珠菌种子被鉴定为顽固的，并成功保存冷冻。在冷冻保存过程中抑制ROS的积累可能是一种潜在的策略，以提高冷冻保存后顽固种子的存活率。
Nymphaea caerulea is a valuable freshwater aquatic plant, not only because of its ornamental value but also its extractions for chemical and medical uses. It is necessary to store its seeds as backup resources. Cryopreservation is the reliable, cost-effective method for long-term preservation of botanical genetic banks, especially for recalcitrant plants. In this study, we demonstrated that due to unable to tolerate desiccation and low-temperature, N. caerulea is recalcitrant. Since viability was lost before 6 months storage, N. caerulea seeds were not appropriate to long-term store by traditional storage method. The only way to long-term store N. caerulea seeds is cryopreservation. However, Plant Vitrification Solution 3 (PVS3; 50% w/v sucrose, 50% w/v glycerol), a commonly used plant vitrification solution, was ineffective on N. caerulea seeds cryopreservation. The maximum survival rate of cryopreserved seeds treated by PVS3 was only 23%. Oxidative stress induced by reactive oxygen species (ROS) accumulation within seeds was the reason to cause inefficiency of PVS3 treated seeds cryopreservation. We developed a new plant vitrification solution (PVS+), by adding glutathione (GSH) into PVS3. PVS+ rescued the inefficiency by decreasing ROS accumulation and elevated survival rate of cryopreserved seeds to 97%. Our results showed some inefficiencies of plant tissue cryopreservation may be caused by ROS-induced oxidative stress. Antioxidants can reduce ROS-induced oxidative stress and improve seeds survival after cryopreservation. In conclusion, N. caerulea seeds were identified as recalcitrant and successfully cryopreserved. Suppressing ROS accumulation during cryopreservation may be a potential strategy to improve survival rate of recalcitrant seeds after cryopreservation.
