Cryopreservation of highbush blueberry, strawberry, and saskatoon using V and D cryo-plate methods and monitoring of multiplication ability of regenerated shoots
{"title":"Cryopreservation of highbush blueberry, strawberry, and saskatoon using V and D cryo-plate methods and monitoring of multiplication ability of regenerated shoots","authors":"","doi":"10.1007/s11627-023-10399-5","DOIUrl":null,"url":null,"abstract":"<h3>Abstract</h3> <p>The purpose of this work was to apply V and D cryo-plate methods for cryopreservation of <em>Vaccinium corymbosum</em> ʻToroʼ, <em>Fragaria</em> × <em>ananassa</em> ʻCleryʼ, and <em>Amelanchier alnifolia</em> (Nutt.) M. Roem. and to monitor the multiplication capacity of shoots regenerated from cryopreserved explants. Shoot tips pre-cultured for 1 d at 23°C in the dark on medium containing 0.3 M sucrose were used as explants. Loading was performed in a solution containing 2 M glycerol and 0.8 M sucrose (30 min at room temperature). In the V cryo-plate, dehydration was carried out at room temperature (20 to 50 min) using the following plant vitrification solutions: original PVS2, 90% PVS2 solution, and PVS3. Regarding the D cryo-plate, dehydration was performed in closed glass containers over silica gel for 2, 2.5, or 3 h. In both protocols, rewarming was carried out in a 1.0 M sucrose solution (15 min at 25°C). Regenerated shoots were multiplied and multiplication parameters were monitored after the second subculture. Using the V cryo-plate method, the highest regrowth in highbush blueberry was obtained following 50-min treatment with all three VSs (61.7 to 80.9%). The D cryo-plate method was even more suitable with maximum regrowth of 89.4% achieved after 2.5 h of desiccation. For strawberry, 62.5% was the highest regrowth recorded using PVS3-based V cryo-plate method while 83.3% of regrowth was observed using D cryo-plate protocol. Regrowth of saskatoon reached a maximum of 50% after 50-min treatment with PVS3 while it did not exceed 40% in other treatments. By the second subculture, shoots regenerated from cryopreserved explants regained and even exceeded the multiplication capacity of shoots regenerated from non-cryopreserved explants. This study is the first to present the successful application of the V cryo-plate method in highbush blueberry, as well as the utilization of both V and D cryo-plate methods in saskatoon.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"5 1","pages":""},"PeriodicalIF":2.2000,"publicationDate":"2024-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"In Vitro Cellular & Developmental Biology - Plant","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1007/s11627-023-10399-5","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
The purpose of this work was to apply V and D cryo-plate methods for cryopreservation of Vaccinium corymbosum ʻToroʼ, Fragaria × ananassa ʻCleryʼ, and Amelanchier alnifolia (Nutt.) M. Roem. and to monitor the multiplication capacity of shoots regenerated from cryopreserved explants. Shoot tips pre-cultured for 1 d at 23°C in the dark on medium containing 0.3 M sucrose were used as explants. Loading was performed in a solution containing 2 M glycerol and 0.8 M sucrose (30 min at room temperature). In the V cryo-plate, dehydration was carried out at room temperature (20 to 50 min) using the following plant vitrification solutions: original PVS2, 90% PVS2 solution, and PVS3. Regarding the D cryo-plate, dehydration was performed in closed glass containers over silica gel for 2, 2.5, or 3 h. In both protocols, rewarming was carried out in a 1.0 M sucrose solution (15 min at 25°C). Regenerated shoots were multiplied and multiplication parameters were monitored after the second subculture. Using the V cryo-plate method, the highest regrowth in highbush blueberry was obtained following 50-min treatment with all three VSs (61.7 to 80.9%). The D cryo-plate method was even more suitable with maximum regrowth of 89.4% achieved after 2.5 h of desiccation. For strawberry, 62.5% was the highest regrowth recorded using PVS3-based V cryo-plate method while 83.3% of regrowth was observed using D cryo-plate protocol. Regrowth of saskatoon reached a maximum of 50% after 50-min treatment with PVS3 while it did not exceed 40% in other treatments. By the second subculture, shoots regenerated from cryopreserved explants regained and even exceeded the multiplication capacity of shoots regenerated from non-cryopreserved explants. This study is the first to present the successful application of the V cryo-plate method in highbush blueberry, as well as the utilization of both V and D cryo-plate methods in saskatoon.
期刊介绍:
Founded in 1965, In Vitro Cellular & Developmental Biology - Plant is the only journal devoted solely to worldwide coverage of in vitro biology in plants. Its high-caliber original research and reviews make it required reading for anyone who needs comprehensive coverage of the latest developments and state-of-the-art research in plant cell and tissue culture and biotechnology from around the world.