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冷冻保存方法结合生化分析对宝石兰腋芽的影响

Effect of Cryopreservation Method Supported with Biochemical Analyses in the Axillary Bud of Jewel Orchid, .

作者信息

Burkhan Hazirah, Rajan Kirutika Selva, Appalasamy Suganthi, Poobathy Ranjetta, Chew Bee Lynn, Mariappan Vanitha, Subramaniam Sreeramanan

机构信息

School of Biological Sciences, Universiti Sains Malaysia (USM), Georgetown 11800, Penang, Malaysia.

Department of Natural Resource and Sustainability, Faculty of Earth Science, Universiti Malaysia Kelantan (UMK), Locked Bag No. 100, Jeli 17600, Kelantan, Malaysia.

出版信息

Plants (Basel). 2022 Mar 25;11(7):879. doi: 10.3390/plants11070879.

DOI:10.3390/plants11070879
PMID:35406859
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9002730/
Abstract

This study investigated conserving an endangered terrestrial jewel orchid using in vitro grown axillary buds. Excised segments of axillary buds (4-5 mm in length) were precultured on a modified Murashige and Skoog (MS) medium supplemented with 0.2 M sucrose for 24 h and osmoprotected in a loading solution for 20 min. Then, axillary buds were dehydrated in plant vitrification solution 2 (PVS2) for 10 min at 0 °C and incubated in liquid nitrogen for 1 h. Subsequently, axillary buds were rewarmed rapidly by dilution solution and transferred to a growth recovery medium supplemented with 0.05 µM melatonin, which led to an improved survival chance (16.67%) for cryopreserved . The osmotic stress and the overproduction of reactive oxygen species (ROS) during cryopreservation stages may result in cryoinjuries and poor survival as increased levels of proline (5.51 µmol/g), catalase (85.64 U/g), peroxidase (565.37 U/g), and ascorbate peroxidase activities (12.19 U/g) were detected after dehydration, preculture, rewarming, and loading stage, respectively. Results obtained from this study indicate that further experimental designs which apply different PVS and exogenous antioxidants are needed for improved survival and regrowth of

摘要

本研究调查了利用离体培养的腋芽对一种濒危地生宝石兰进行保存的方法。将长度为4 - 5毫米的腋芽切段在添加了0.2 M蔗糖的改良Murashige和Skoog（MS）培养基上预培养24小时，并在装载溶液中进行渗透保护20分钟。然后，将腋芽在植物玻璃化溶液2（PVS2）中于0°C脱水10分钟，并在液氮中孵育1小时。随后，通过稀释溶液使腋芽快速复温，并转移到添加了0.05 µM褪黑素的生长恢复培养基中，这使得冷冻保存的腋芽存活几率提高（16.67%）。在脱水、预培养、复温和装载阶段后分别检测到脯氨酸（5.51 µmol/g）、过氧化氢酶（85.64 U/g）、过氧化物酶（565.37 U/g）和抗坏血酸过氧化物酶活性（12.19 U/g）水平升高，这表明冷冻保存阶段的渗透胁迫和活性氧（ROS）过量产生可能导致冷冻损伤和低存活率。本研究所得结果表明，需要进一步设计应用不同PVS和外源抗氧化剂的实验，以提高[具体植物名称未完整给出]的存活率和再生能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/8d6b2cdbf702/plants-11-00879-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/40298fe2e124/plants-11-00879-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/b0d1c7dd1d1c/plants-11-00879-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/7e903bbab24c/plants-11-00879-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/8189c810f467/plants-11-00879-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/7f7891f05631/plants-11-00879-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/97536ba89058/plants-11-00879-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/65bba21c40d0/plants-11-00879-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/49d015114578/plants-11-00879-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/8d6b2cdbf702/plants-11-00879-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/40298fe2e124/plants-11-00879-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/b0d1c7dd1d1c/plants-11-00879-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/7e903bbab24c/plants-11-00879-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/8189c810f467/plants-11-00879-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/7f7891f05631/plants-11-00879-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/97536ba89058/plants-11-00879-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/65bba21c40d0/plants-11-00879-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/49d015114578/plants-11-00879-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/733e/9002730/8d6b2cdbf702/plants-11-00879-g009.jpg

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ROS-induced oxidative stress in plant cryopreservation: occurrence and alleviation.植物冷冻保存中 ROS 诱导的氧化应激：发生与缓解。
Planta. 2021 Nov 20;254(6):124. doi: 10.1007/s00425-021-03784-0.
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Looking for a way forward for the cryopreservation of orchid diversity.寻找兰花多样性冷冻保存的前进之路。
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Plants (Basel). 2020 Nov 24;9(12):1634. doi: 10.3390/plants9121634.
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Droplet-vitrification of Broga Blue orchid: Role of ascorbic acid on the antioxidant system and genetic fidelity assessments via RAPD and SCoT markers.布罗加蓝花石斛的玻璃化法超低温保存：抗坏血酸在抗氧化系统中的作用以及通过随机扩增多态性DNA（RAPD）和目标起始密码子多态性（SCoT）标记进行的遗传稳定性评估
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