PDF(Pubmed)
Abstract：
Cryopreservation is an important technique used in the conservation of various plant tissues. This study proposes a cryopreservation method for the long-term conservation of eastern bracken fern gametophytes (Pteridium aquilinum var. latiusculum). Encapsulation-dehydration of the gametophytes was performed, and the exogenous sucrose and abscisic acid (ABA) preculture conditions were investigated. Gametophytes are sensitive to dehydration and drying, and the following treatment conditions were applied: encapsulation by alginate containing 0.75 M sucrose, 18-h loading treatment with 0.75 M sucrose, and 6-h drying treatment. The survival rate following cryopreservation was determined. The water content of < 27.5% in the alginate beads after dehydration and drying was found to be appropriate for ensuring survival. Additionally, performing an exogenous sucrose and ABA preculture was essential before encapsulation to achieve a survival of ≥ 90%. The high stress induced by cryopreservation and exogenous preculture regulated the expression of PaSuSy, PaLEA14, and PaABI1b and the endogenous ABA content. In eastern bracken gametophytes, ABI1 appears to be a negative regulator of ABA signaling. These results indicate that the encapsulation-dehydration method is effective for the long-term conservation of eastern bracken fern gametophytes, and exogenous preculture alleviates abiotic stress and increases the survival rate.
摘要：
冷冻保存是用于保护各种植物组织的重要技术。本研究提出了一种冷冻保存方法，用于长期保存东部的蕨类植物配子体（Pteridiumaquilinumvar。latiusculum).进行配子体的包封-脱水，并研究了外源蔗糖和脱落酸（ABA）的预培养条件。配子体对脱水和干燥敏感，并应用以下处理条件:用含有0.75M蔗糖的藻酸盐封装，用0.75M蔗糖进行18小时的装载处理，和6小时的干燥处理。测定冷冻保存后的存活率。发现脱水和干燥后藻酸盐珠粒中<27.5%的水含量适合于确保存活。此外,在封装之前进行外源蔗糖和ABA预培养以实现≥90％的存活率是必不可少的。低温保存和外源预培养诱导的高胁迫对PaSuSy表达的调控,PaLEA14、PaABI1b和内源ABA含量。在东部蕨菜配子体中，ABI1似乎是ABA信号的负调节因子。这些结果表明，包封-脱水方法对于长期保存东部蕨类植物是有效的。外源预培养减轻了非生物胁迫，提高了存活率。
