{"title":"Cryopreservation of Arum palaestinum plant callus as a strategy for mitigating extinction risks","authors":"","doi":"10.1016/j.cpb.2024.100402","DOIUrl":null,"url":null,"abstract":"<div><div><em>Arum palaestinum</em> is a wild perennial plant commonly known as \"Al-Loof\" in Jordan. Due to overharvesting, climate change, and increasing demand, its natural populations are threatened with extinction. Cryopreservation, an effective method for conserving plant material at ultra-low temperatures, is explored for <em>A. palaestinum</em> calli. We investigated the applicability of encapsulation-vitrification (using different plant vitrification solutions (PVS) and incubation times), encapsulation-dehydration (using sucrose or sorbitol at different concentrations and dehydration times), and the v-cryoplate (using different pre-culture times and temperatures) techniques. In the encapsulation-vitrification experiment, a notable 82.4 % regrowth rate was achieved by desiccating calli in plant vitrification solution 2 (PVS2) for 10 minutes at 25 °C. The encapsulation-dehydration technique resulted in an 82.6 % regrowth rate by incubating calli for one day in low sucrose levels (0.1 M sucrose) following one hour of air dehydration, where the moisture content of the beads was 30 %. The moisture content of the beads decreased from 81 % before chemical and air dehydration to 71 % after 0 hours of air dehydration combined with chemical dehydration using 0.1 M sucrose or sorbitol. It further dropped to 30–34 % after one day of chemical dehydration with 0.1 M sucrose and 1 hour of air dehydration. The v-cryoplate technique successfully conserved calli, showing impressive survival and regrowth percentages (96.8 %) when the callus was pre-cultured with 0.3 M sucrose for three days at 5 °C. Temperature during pre-culture significantly influenced regrowth percentages in the v-cryoplate technique. The study establishes promising cryopreservation protocols for <em>A. palaestinum</em> calli, offering a means to conserve germplasm and contribute to environmental and biodiversity protection by reintroducing endangered plants to their native habitats.</div></div>","PeriodicalId":38090,"journal":{"name":"Current Plant Biology","volume":null,"pages":null},"PeriodicalIF":5.4000,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Plant Biology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2214662824000847","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
引用次数: 0
Abstract
Arum palaestinum is a wild perennial plant commonly known as "Al-Loof" in Jordan. Due to overharvesting, climate change, and increasing demand, its natural populations are threatened with extinction. Cryopreservation, an effective method for conserving plant material at ultra-low temperatures, is explored for A. palaestinum calli. We investigated the applicability of encapsulation-vitrification (using different plant vitrification solutions (PVS) and incubation times), encapsulation-dehydration (using sucrose or sorbitol at different concentrations and dehydration times), and the v-cryoplate (using different pre-culture times and temperatures) techniques. In the encapsulation-vitrification experiment, a notable 82.4 % regrowth rate was achieved by desiccating calli in plant vitrification solution 2 (PVS2) for 10 minutes at 25 °C. The encapsulation-dehydration technique resulted in an 82.6 % regrowth rate by incubating calli for one day in low sucrose levels (0.1 M sucrose) following one hour of air dehydration, where the moisture content of the beads was 30 %. The moisture content of the beads decreased from 81 % before chemical and air dehydration to 71 % after 0 hours of air dehydration combined with chemical dehydration using 0.1 M sucrose or sorbitol. It further dropped to 30–34 % after one day of chemical dehydration with 0.1 M sucrose and 1 hour of air dehydration. The v-cryoplate technique successfully conserved calli, showing impressive survival and regrowth percentages (96.8 %) when the callus was pre-cultured with 0.3 M sucrose for three days at 5 °C. Temperature during pre-culture significantly influenced regrowth percentages in the v-cryoplate technique. The study establishes promising cryopreservation protocols for A. palaestinum calli, offering a means to conserve germplasm and contribute to environmental and biodiversity protection by reintroducing endangered plants to their native habitats.
Arum palaestinum 是一种多年生野生植物,在约旦通常被称为 "Al-Loof"。由于过度采摘、气候变化和需求增加,其自然种群正面临灭绝的威胁。低温保存是一种在超低温下保存植物材料的有效方法,我们对 A. palaestinum 的胼胝体进行了探索。我们研究了封装-玻璃化(使用不同的植物玻璃化溶液(PVS)和培养时间)、封装-脱水(使用不同浓度的蔗糖或山梨醇和脱水时间)以及 V 型冻存(使用不同的预培养时间和温度)技术的适用性。在封装-玻璃化实验中,将胼胝体置于植物玻璃化溶液 2(PVS2)中,在 25 °C 下干燥 10 分钟,再生率达到 82.4%。通过封装-脱水技术,将胼胝体在低浓度蔗糖(0.1 M 蔗糖)中培养一天,然后在空气中脱水一小时,珠子的含水量为 30%,再生率为 82.6%。在使用 0.1 M 蔗糖或山梨醇进行化学脱水和空气脱水 0 小时后,珠子的含水量从化学脱水和空气脱水前的 81% 降至 71%。在使用 0.1 M 蔗糖进行化学脱水一天和空气脱水 1 小时后,水分含量进一步降至 30-34%。v-cryoplate 技术成功地保存了胼胝体,在 5 °C 下用 0.3 M 蔗糖预培养三天后,胼胝体的存活率和再生率(96.8%)令人印象深刻。预培养过程中的温度对 V 型干细胞技术中的再生率有显著影响。该研究为 A. palaestinum 胼胝体建立了前景广阔的低温保存方案,提供了一种保存种质资源的方法,并通过将濒危植物重新引入其原生栖息地,为环境和生物多样性保护做出了贡献。
期刊介绍:
Current Plant Biology aims to acknowledge and encourage interdisciplinary research in fundamental plant sciences with scope to address crop improvement, biodiversity, nutrition and human health. It publishes review articles, original research papers, method papers and short articles in plant research fields, such as systems biology, cell biology, genetics, epigenetics, mathematical modeling, signal transduction, plant-microbe interactions, synthetic biology, developmental biology, biochemistry, molecular biology, physiology, biotechnologies, bioinformatics and plant genomic resources.