通过玻璃化液滴法对欧洲百合茎尖进行超低温保存及其生理稳定性评估

Cryopreservation of Lilium martagon l. meristems by droplet-vitrification and evaluation of their physiological stability.

作者信息

Urbaniec-Kiepura M, Bach A

机构信息

Department of Ornamental Plants, University of Agriculture in Krakow, Krakow, Poland.

出版信息

Cryo Letters. 2017 Mar/Apr;38(2):78-89.

PMID:28534050

Abstract

BACKGROUND

The aim of the present study was to compare different strategies for cryopreservation of martagon lily meristems and to evaluate the physiological status of the regenerants.

MATERIALS AND METHODS

The bulblets were stored at 5 degree C or 20 degree C and pretreated with 3% or 6% sucrose prior to droplet-vitrification. The meristems were then assessed for their survival and regeneration. Their photochemical activity was investigated using a Photosynthesis Yield Analyzer MINI PAM 2000 Portable Chlorophyll Fluorometer and their photosynthesis oxygen production was evaluated with a Plant Vital 5030 device.

RESULTS

The plant material stored at 5 degree C on medium containing 3% sucrose exhibited lower survival (40.8%) and regeneration (75%) of meristems following cryopreservation compared with material stored at 20 degree C on medium containing 3% sucrose, for which survival was 65% and regeneration 87%. Treatment of lily meristems for 30 min with PVS2 yielded high survival and regeneration. The implemented cryopreservation protocol did not induce any physiological changes in regenerants. Chlorophyll fluorescence (Fv/Fm) was 0.822 for cryopreserved samples (+LN) and 0.824 for non-cryopreserved ones (-LN). Photosynthetic oxygen production (KphA) was 1.531 (+LN) and 1.410 (-LN).

CONCLUSION

Droplet-vitrification seems to be an effective method for cryopreservation of martagon lily meristems with the aim of its ex situ protection.

摘要

背景

本研究的目的是比较铃兰分生组织冷冻保存的不同策略，并评估再生植株的生理状态。

材料与方法

将小鳞茎保存在5℃或20℃，并在玻璃化滴冻前用3%或6%的蔗糖进行预处理。然后评估分生组织的存活和再生情况。使用光合作用产量分析仪MINI PAM 2000便携式叶绿素荧光计研究其光化学活性，并用Plant Vital 5030设备评估其光合作用产氧量。

结果

与保存在含有3%蔗糖的培养基上、温度为20℃的材料相比，保存在含有3%蔗糖的培养基上、温度为5℃的植物材料在冷冻保存后分生组织的存活率（40.8%）和再生率（75%）较低，前者的存活率为65%，再生率为87%。用PVS2处理铃兰分生组织30分钟可获得较高的存活率和再生率。实施的冷冻保存方案未在再生植株中引起任何生理变化。冷冻保存样品（+LN）的叶绿素荧光（Fv/Fm）为0.822，未冷冻保存样品（-LN）为0.824。光合产氧量（KphA）分别为1.531（+LN）和1.410（-LN）。