A ten-year retrospective on the efficacy of droplet vitrification for cryobanking of Allium ramosum L. germplasm.

IF 1 4区生物学 Q3 BIOLOGY

Cryo letters Pub Date : 2025-03-01

S Chander, R Gowthami, R Pandey, D A Deepak, A Agrawal

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引用次数: 0

Abstract

Background: Allium ramosum is an important member of the genus Allium, which is commonly known as Chinese chive or fragrant-flowered garlic. Conserving the genetic diversity of different species of Allium is crucial, and cryopreservation has emerged as an important strategy for long-term conservation of alliums.

Objective: To develop a reliable protocol for the cryoconservation of A. ramosum shoot bases.

Materials and methods: Different parameters, viz. (a) cold-hardening (5 degree C for 16/8 h photoperiod), (b) PVS2 dehydration (0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 and 60 min), (c) pregrowth medium (MM3: MS + 0.1 mg/L NAA + 0.5 mg/L 2iP + 10 mg/L spermidine + 3% sucrose; MM10: MS + 0.1 mg/L NAA + 0.5 mg/l 2iP + 10 mg/L spermidine + 10% sucrose) and (d) preculture duration (1, 2, 3, 4 and 5 days) were tested using a vitrification technique.

Results: Shoot bases excised from 4-wk old in vitro cultures that had been cold-hardened at 5 degree C (16/8 h photoperiod) and precultured on MM10 with 10% sucrose at 5 degree C for 3 days resulted in highest post-thaw regrowth of 43% after conventional vitrification. However, when droplet-vitrification was used, post-thaw regrowth was increased to 77%. Retesting of shoot bases after 10 years of cryobanking, revealed no significant difference in the post-thaw regrowth of A. ramosum.

Conclusion: This is the first report of the long-term cryopreservation of A. ramosum shoot bases using vitrification and droplet-vitrification techniques. https://doi.org/10.54680/fr25210110512.

本刊更多论文

液滴玻璃化法冷冻保存小葱种质的十年回顾。

背景：ramosum Allium ramosum是Allium属的重要成员，通常被称为韭菜或香蒜。保存不同种类葱属植物的遗传多样性至关重要，低温保存已成为长期保存葱属植物的重要策略。目的：建立一套可靠的冷保存方法。材料和方法：不同的参数，即(a)冷硬化（5℃，16/8 h光周期），(b) PVS2脱水（0、5、10、15、20、25、30、35、40、45、50、55和60 min）， (C)生长前培养基(MM3: MS + 0.1 mg/L NAA + 0.5 mg/L 2iP + 10 mg/L亚精胺+ 3%蔗糖；采用玻璃化技术检测MM10: MS + 0.1 mg/L NAA + 0.5 mg/L 2iP + 10 mg/L亚精胺+ 10%蔗糖)和(d)预培养时间（1、2、3、4和5天）。结果：从4周龄的离体培养物中取出茎基，在5℃（16/8小时光周期）下冷硬化，并在5℃下用10%蔗糖在MM10上预培养3天，常规玻璃化后解冻后再生率最高，为43%。然而，当使用液滴玻璃化时，解冻后再生率提高到77%。冷冻贮藏10年后，重新测试的芽基显示，冻融后再生无显著差异。结论：本研究首次采用玻璃化和液滴玻璃化技术对沙菖蒲茎底进行了长期冷冻保存。https://doi.org/10.54680/fr25210110512。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cryo letters 生物-生理学

CiteScore

1.80

自引率

10.00%

发文量

审稿时长

1 months

期刊介绍： A bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.