Gentiana kurroo Royle, a critically endangered medicinal plant species of India is conserved as in vitro slow growing cultures in the In Vitro Genebank (IVGB) at ICAR-National Bureau of Plant Genetic Resources (NBPGR), New Delhi, India. Shoot tips (about 1 mm in length) excised from 8-weeks-old stock cultures subjected to cold hardening for four weeks at 8 °C (dark) were precultured on Murashige and Skoog (MS) medium supplemented with 5% Dimethylsulfoxide (DMSO) at 8 °C (dark) for two days. Thereafter, shoot tips were dehydrated with PVS2 solution (30 min) at 0 °C and cryopreserved using conventional vitrification (V) and droplet-vitrification (DV) techniques. Average post-thaw regeneration after 4 weeks was ~ 35% by V technique and ~ 60% by DV in all the three accessions tested. Genetic stability assessment on the basis of 40 Inter Simple Sequence Repeats and 30 Expressed Sequence Tagged-Simple Sequence Repeats markers revealed no variation between in vitro-cryopreserved plants and controls, in any of the genotypes. Thus, standardized DV method has the potential for long-term conservation of germplasm of G. kurroo and the three accessions are now safely cryobanked at ICAR-NBPGR.

秦ianaRoyle是印度极度濒危的药用植物，在印度新德里的ICAR国家植物遗传资源局（NBPGR）的体外基因库（IVGB）中作为体外缓慢生长的培养物得以保存。从8周龄的原种培养物中切下的茎尖（长度约1毫米）在8°C（黑暗）下进行了4年的冷硬化处理，然后在补充了5％二甲基亚砜（DMSO）的Murashige和Skoog（MS）培养基上进行了预培养在8°C（黑暗）下放置两天。此后，将茎尖在0°C下用PVS2溶液脱水（30分钟），并使用常规玻璃化（V）和液滴玻璃化（DV）技术进行低温保存。在所有测试的三个种质中，用V技术测得的4周后的平均解冻后再生率约为35％，DV约为60％。基于40个内部简单序列重复和30个表达的序列标签-简单序列重复标记的遗传稳定性评估显示，在任何基因型中，体外冷冻保存的植物和对照之间均无差异。因此，标准化的DV方法具有长期保存种质资源的潜力。G. kurroo和三个种质现已安全地保存在ICAR-NBPGR中。