M S Swaminathan Research Foundation Community Agrobiodiversity Centre, Kalpetta, Wayanad, Kerala, India.
Science, Innovation and Diagnostics division and Bio-Agriculture, Avesthagen Ltd, Bangalore, India, Affiliated to University of Mysore, Karnataka-570005, India.
Cryo Letters. 2021 May-Jun;42(3):168-177.
Cryopreservation is a reliable and economical method for the long-term ex situ conservation of valuable genetic resources.
The present study focuses on establishing novel regeneration strategies and on assessing various cryogenic methods using nodal explants/shoot apices and on developing in vitro technologies for germplasm conservation of Dioscorea prazeri.
Pre-treatment, growth regulators, temperature conditions, treatment period for recovery and growth of explants were optimized and various germplasm conservation methods were conducted to attain the conservation and mass multiplication of the endangered therapeutic plant. The plants regenerated from vitrified tissues were evaluated for physiological stability through morphological characteristics, genetic stability using RAPD analysis and with key metabolites for biochemical characterization.
An optimized vitrification method resulted in a regeneration level of 92 ± 2 %, whereas a method comprising encapsulation dehydration resulted in 75 ± 2 % regeneration. In contrast, only a 38 ± 2 % regeneration was achieved using an encapsulation vitrification method.
Vitrification-based procedures significantly improve cryopreservation survival and can be successfully employed for the long-term conservation of Dioscorea species and, potentially, other medicinal plants.
冷冻保存是长期体外保存有价值遗传资源的可靠且经济的方法。
本研究旨在建立新的再生策略,并使用节间外植体/茎尖评估各种冷冻方法,并开发用于穿龙薯蓣种质保存的体外技术。
优化预处理、生长调节剂、温度条件、恢复和外植体生长的处理期,并采用各种种质保存方法来实现濒危治疗植物的保存和大量繁殖。通过形态特征、RAPD 分析和关键代谢物进行生化特性鉴定来评估玻璃化组织再生植株的生理稳定性。
优化的玻璃化方法可获得 92±2%的再生水平,而包含包埋脱水的方法可获得 75±2%的再生水平。相比之下,仅使用包埋玻璃化方法可获得 38±2%的再生水平。
基于玻璃化的程序可显著提高冷冻保存的存活率,可成功用于穿龙薯蓣属和其他药用植物的长期保存。