Cryopreservation and assessment of genetic fidelity Acorus calamus Linn., an endangered medicinal plant.

IF 1 4区生物学 Q3 BIOLOGY

Cryo letters Pub Date : 2024-03-01

K Parasher, C Sharma, S Sharma, EmptyYN Y Shrishti, P Mukherjee

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引用次数: 0

Abstract

Background: Acorus calamus Linn. is a medicinally valuable monocot plant belonging to the family Acoraceae. Over-exploitation and unscientific approach towards harvesting to fulfill an ever-increasing demand have placed it in the endangered list of species.

Objective: To develop vitrification-based cryopreservation protocols for A. calamus shoot tips, using conventional vitrification and V cryo-plate.

Materials and methods: Shoot tips (2 mm in size) were cryopreserved with the above techniques by optimizing various parameters such as preculture duration, sucrose concentration in the preculture medium, and PVS2 dehydration time. Regenerated plantlets obtained post-cryopreservation were evaluated by random amplified polymorphic DNA (RAPD) to test their genetic fidelity.

Results: The highest regrowth of 88.3% after PVS2 exposure of 60 min was achieved with V cryo-plate as compared to 75% after 90 min of PVS2 exposure using conventional vitrification. After cryopreservation, shoot tips developed into complete plantlets in 28 days on regrowth medium (0.5 mg/L BAP, 0.3 mg/L GA₃, and 0.3 mg/L ascorbic acid). RAPD analysis revealed 100% monomorphism in all cryo-storage derived regenerants and in vitro donor (120-days-old) plants.

Conclusion: Shoot tips of A. calamus that were cryopreserved had 88.3% regrowth using V cryo-plate technique and the regerants retained genetic fidelity. https://doi.org/10.54680/fr24210110412.

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濒危药用植物石菖蒲（Acorus calamus Linn.

背景：石菖蒲（Acorus calamus Linn.）是一种具有药用价值的单子叶植物，属于金合欢科。为了满足日益增长的需求，过度开发和不科学的采收方法已将其列入濒危物种名单：利用传统玻璃化技术和 V 型低温板，开发基于玻璃化技术的菖蒲嫩枝尖低温保存方案：通过优化预培养时间、预培养基中的蔗糖浓度和 PVS2 脱水时间等各种参数，采用上述技术对嫩枝尖（2 毫米大小）进行冷冻保存。通过随机扩增多态性 DNA（RAPD）对冷冻保存后获得的再生小植株进行评估，以检验其遗传保真度：结果：V 型低温板在 PVS2 暴露 60 分钟后的再生率最高，达到 88.3%，而传统玻璃化法在 PVS2 暴露 90 分钟后的再生率为 75%。冷冻保存后，芽尖在再生培养基（0.5 毫克/升 BAP、0.3 毫克/升 GA3 和 0.3 毫克/升抗坏血酸）上 28 天后发育成完整的小植株。RAPD 分析显示，所有低温贮藏的再生植株和离体供体（120 天）植株的单形性均为 100%：https://doi.org/10.54680/fr24210110412。

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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.