Conservation of Green and White Ash Germplasm Using the Cryopreservation of Embryogenic Cultures

Green ash (<i>Fraxinus pennsylvanica</i>) and white ash (<i>F. americana</i>) populations are currently experiencing major declines across their native ranges in North America due to infestation by the exotic insect pest emerald ash borer (<i>Agrilus planipennis</i&g...

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Main Authors: Mason Richins, Cristian Montes, Scott Merkle
Format: Article
Language:English
Published: MDPI AG 2024-01-01
Series:Plants
Subjects:
Online Access:https://www.mdpi.com/2223-7747/13/3/352
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author Mason Richins
Cristian Montes
Scott Merkle
author_facet Mason Richins
Cristian Montes
Scott Merkle
author_sort Mason Richins
collection DOAJ
description Green ash (<i>Fraxinus pennsylvanica</i>) and white ash (<i>F. americana</i>) populations are currently experiencing major declines across their native ranges in North America due to infestation by the exotic insect pest emerald ash borer (<i>Agrilus planipennis</i>). The development of a reliable method for the long-term storage of green and white ash germplasm in the form of embryogenic cultures using cryopreservation would be a considerable aid to ash conservation efforts. We compared recovery percentages of cryopreserved green and white ash embryogenic cultures using vitrification versus slow cooling methods. Three Plant Vitrification Solution 2 (PVS2) exposure durations (40, 60, and 80 min) for vitrification and three DMSO concentrations (5%, 10%, and 15%) for slow cooling were tested for their effects on the percentage of cultures that regrew following cryostorage. Vitrification resulted in a higher overall culture recovery percentage (91%) compared to cultures that were cryostored using the slow cooling approach (39%), and a more rapid initiation of regrowth (5 days versus 2–3 weeks) resulted. Recovery from cryostorage by cultures using the slow cooling approach varied significantly (<i>p</i> < 0.05) between experiments and with genotype (<i>p</i> < 0.05). The recovery of vitrified tissue from cryostorage did not vary with genotype, species, or PVS2 exposure duration (<i>p</i> > 0.05). The vitrification cryopreservation protocol provides a reliable and versatile alternative to the traditional slow cooling method, strengthening our ability to preserve valuable ash germplasm for conservation and restoration.
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spelling doaj.art-d82aac107ab94eb4a28ac67c7f8603df2024-02-09T15:20:09ZengMDPI AGPlants2223-77472024-01-0113335210.3390/plants13030352Conservation of Green and White Ash Germplasm Using the Cryopreservation of Embryogenic CulturesMason Richins0Cristian Montes1Scott Merkle2Warnell School of Forestry and Natural Resources, University of Georgia, Athens, GA 30602, USAWarnell School of Forestry and Natural Resources, University of Georgia, Athens, GA 30602, USAWarnell School of Forestry and Natural Resources, University of Georgia, Athens, GA 30602, USAGreen ash (<i>Fraxinus pennsylvanica</i>) and white ash (<i>F. americana</i>) populations are currently experiencing major declines across their native ranges in North America due to infestation by the exotic insect pest emerald ash borer (<i>Agrilus planipennis</i>). The development of a reliable method for the long-term storage of green and white ash germplasm in the form of embryogenic cultures using cryopreservation would be a considerable aid to ash conservation efforts. We compared recovery percentages of cryopreserved green and white ash embryogenic cultures using vitrification versus slow cooling methods. Three Plant Vitrification Solution 2 (PVS2) exposure durations (40, 60, and 80 min) for vitrification and three DMSO concentrations (5%, 10%, and 15%) for slow cooling were tested for their effects on the percentage of cultures that regrew following cryostorage. Vitrification resulted in a higher overall culture recovery percentage (91%) compared to cultures that were cryostored using the slow cooling approach (39%), and a more rapid initiation of regrowth (5 days versus 2–3 weeks) resulted. Recovery from cryostorage by cultures using the slow cooling approach varied significantly (<i>p</i> < 0.05) between experiments and with genotype (<i>p</i> < 0.05). The recovery of vitrified tissue from cryostorage did not vary with genotype, species, or PVS2 exposure duration (<i>p</i> > 0.05). The vitrification cryopreservation protocol provides a reliable and versatile alternative to the traditional slow cooling method, strengthening our ability to preserve valuable ash germplasm for conservation and restoration.https://www.mdpi.com/2223-7747/13/3/352<i>Fraxinus americana</i><i>Fraxinus pennsylvanica</i>cryostoragesomatic embryogenesisemerald ash borerin vitro culture
spellingShingle Mason Richins
Cristian Montes
Scott Merkle
Conservation of Green and White Ash Germplasm Using the Cryopreservation of Embryogenic Cultures
Plants
<i>Fraxinus americana</i>
<i>Fraxinus pennsylvanica</i>
cryostorage
somatic embryogenesis
emerald ash borer
in vitro culture
title Conservation of Green and White Ash Germplasm Using the Cryopreservation of Embryogenic Cultures
title_full Conservation of Green and White Ash Germplasm Using the Cryopreservation of Embryogenic Cultures
title_fullStr Conservation of Green and White Ash Germplasm Using the Cryopreservation of Embryogenic Cultures
title_full_unstemmed Conservation of Green and White Ash Germplasm Using the Cryopreservation of Embryogenic Cultures
title_short Conservation of Green and White Ash Germplasm Using the Cryopreservation of Embryogenic Cultures
title_sort conservation of green and white ash germplasm using the cryopreservation of embryogenic cultures
topic <i>Fraxinus americana</i>
<i>Fraxinus pennsylvanica</i>
cryostorage
somatic embryogenesis
emerald ash borer
in vitro culture
url https://www.mdpi.com/2223-7747/13/3/352
work_keys_str_mv AT masonrichins conservationofgreenandwhiteashgermplasmusingthecryopreservationofembryogeniccultures
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