Untitled

Vendrame, Wagner A.; Holliday, C. P.; Montello, Paul; Smith, Dale M.; Merkle, S. A.

doi:10.1023/a:1012237606373

Cited by 21 publications

(7 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this study, the DMSO pretreatment with 0.4 M sorbitol produced the highest percentage of recovery (46%) across all three tested culture lines among the tested DMSO concentrations. A concentration of 0.4 M sorbitol has performed well in other studies, while the optimal DMSO concentration has varied [36]. In that study, PEMs were suspended in the cryoprotectant solution just prior to freezing in an attempt to minimize phytotoxic effects.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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

Richins,

Montes,

Merkle

2024

Plants

View full text Add to dashboard Cite

Green ash (Fraxinus pennsylvanica) and white ash (F. americana) populations are currently experiencing major declines across their native ranges in North America due to infestation by the exotic insect pest emerald ash borer (Agrilus planipennis). 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 (p < 0.05) between experiments and with genotype (p < 0.05). The recovery of vitrified tissue from cryostorage did not vary with genotype, species, or PVS2 exposure duration (p > 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.

show abstract

Section: Discussionmentioning

confidence: 99%

“…The protocol tested in this experiment was adapted from a method reported for the cryostoring embryogenic cultures of Liriodendron tulipifera and Liquidambar spp. [36]. It involved three main stages: preculture, two-step freezing, and thawing.…”

Section: Slow Cooling Experimentsmentioning

confidence: 99%

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

Richins,

Montes,

Merkle

2024

Plants

View full text Add to dashboard Cite

show abstract

“…Means with common letters are not significantly different at P = 0.05 level according to LSD test conditions as these prevent freezing injury and improve post-thawing viability (Mazur 2004). Thus, cellular water content must be reduced either by use of cryoprotectants, such as DMSO (Vendrame et al 2001), polyethylene glycol (PEG) (Hagmann et al 1998), glycerol (Hirata et al 2002) or by subjecting cells to osmotic pretreatment using agents such as sugar or to other compounds that contribute to dehydration (Blakesley et al 1993). Treatment of cells/ tissues with liquid nitrogen stops all metabolic activities at the cellular level, and does not require water, thus avoiding ice formation.…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation of embryogenic cell suspensions of Catharanthus roseus L. (G) Don.

Fatima

Mujib

Nasim

et al. 2009

Plant Cell Tiss Organ Cult

View full text Add to dashboard Cite

An efficient cryopreservation protocol was established for embryogenic cell suspension cultures of Catharanthus roseus. This involved a vitrification-based cryopreservation method wherein embryogenic cells were exposed to a preculture/pretreatment medium prior to their immersion in liquid nitrogen. These cell suspension cultures were first initiated from friable embryogenic callus derived from hypocotyls of C. roseus on a medium containing 4.52 lM 2,4-Dichlorophenoxy acetic acid (2, 4-D). Among different sucrose (0.09-0.6 M) and sorbitol (0.2-0.6 M) levels evaluated during preculture, 0.4 M sucrose promoted highest cellular regrowth. Whereas, among pretreatments Dimethyl sulphoxide (DMSO) (5 or 10%) and glycerol (5 or 10%) at six different levels either alone or in combinations (PT 1-PT 6), the cryopreservation treatment combinations of either 0.4 M sucrose, 5% DMSO, and 5% glycerol (PT-5) or 0.4 M sucrose and 5% DMSO (PT-1) resulted in the highest frequency of viability of embryogenic cultures. However, the PT-1 treatment produced highest number of cell colonies (10.06 ± 0.55) following reculture of cryopreseved cultures. All calluses regrown in an optimized medium, containing 6.62 lM 6-benzyladenine (BA) and 5.37 lM a-naphthaleneacetic acid (NAA), resumed normal growth, and produced somatic embryos similar to those from non-frozen embryogenic cultures. These somatic embryos were converted into regenerated plantlets, and all plantlets exhibited normal morphology.

show abstract

“…Direct immersion in LN of embryogenic explants is frequently used, but positive results have also been reported after slow cooling (Jekkel et al, 1998;Vendrame et al, 2001;Scocchi et al, 2007;Ozudogru et al, 2010;Yu et al, 2022). Rapid thawing approach is employed in most protocols, whereby the cryopreserved tissues are plunged into water at 40-42°C for 2-3 min (Supplementary Material 3).…”

Section: Cryopreservation In Deciduous Forest Speciesmentioning

confidence: 99%

Current status of the cryopreservation of embryogenic material of woody species

Ballesteros,

Martínez,

Sánchez-Romero

et al. 2024

Front. Plant Sci.

View full text Add to dashboard Cite

Cryopreservation, or the storage at liquid nitrogen temperatures (-196°C), of embryogenic cells or somatic embryos allows their long-term conservation without loss of their embryogenic capacity. During the last decade, protocols for cryopreservation of embryogenic material of woody species have been increasing in number and importance. However, despite the large experimental evidence proved in thousands of embryogenic lines, the application for the large-scale conservation of embryogenic material in cryobanks is still limited. Cryopreservation facilitates the management of embryogenic lines, reducing costs and time spent on their maintenance, thus limiting the risk of the appearance of somaclonal variation or contamination. Somatic embryogenesis in combination with cryopreservation is especially useful to preserve the juvenility of lines while the corresponding clones are being field-tested. Hence, when tree performance has been evaluated, selected varieties can be propagated from the cryostock. The traditional method of slow cooling or techniques based on vitrification are mostly applied procedures. For example, slow cooling methods are widely applied to conserve embryogenic lines of conifers. Desiccation based procedures, although simpler, have been applied in a smaller number of species. Genetic stability of the cryopreserved material is supported by multiloci PCR-derived markers in most of the assayed species, whereas DNA methylation status assays showed that cryopreservation might induce some changes that were also observed after prolonged subculture of the embryogenic lines. This article reviews the cryopreservation of embryogenic cultures in conifers, fruit species, deciduous forest species and palms, including a description of the different cryopreservation procedures and the analysis of their genetic stability after storage in liquid nitrogen.

show abstract

Untitled

Cited by 21 publications

References 11 publications

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

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

Cryopreservation of embryogenic cell suspensions of Catharanthus roseus L. (G) Don.

Current status of the cryopreservation of embryogenic material of woody species

Contact Info

Product

Resources

About