Comparing encapsulation-dehydration and droplet-vitrification for cryopreservation of sugarcane (Saccharum spp.) shoot tips

Barraco, Giacoma; Sylvestre, Isabelle; Engelmann, Florent

doi:10.1016/j.scienta.2011.07.003

Cited by 27 publications

(21 citation statements)

References 8 publications

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“…The availability of an efficient and reliable cryogenic protocol yielding a high recovery percentage is a prerequisite for large scale, routine application of cryopreservation [10].…”

Section: Introductionmentioning

confidence: 99%

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Cryopreservation of grapevine (Vitis vinifera L.) in vitro shoot tips

Marković¹,

Chatelet

Sylvestre

et al. 2013

Open Life Sciences

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In this work, we compared the efficiency of encapsulation-dehydration and droplet-vitrification techniques for cryopreserving grapevine (Vitis vinifera L.) cv. Portan shoot tips. Recovery of cryopreserved samples was achieved with both techniques; however, droplet-vitrification, which was used for the first time with grapevine shoot tips, produced higher regrowth. With encapsulationdehydration, encapsulated shoot tips were precultured in liquid medium with progressively increasing sucrose concentrations over a 2-day period (12 h in medium with 0.25, 0.5, 0.75 and 1.0 M sucrose), then dehydrated to 22.28% moisture content (fresh weight). After liquid nitrogen exposure 37.1% regrowth was achieved using 1 mm-long shoot tips and only 16.0% with 2 mm-long shoot tips. With droplet-vitrification, 50% regrowth was obtained following treatment of shoot tips with a loading solution containing 2 M glycerol + 0.4 M sucrose for 20 min, dehydration with half-strength PVS2 vitrification solution (30% (w/v) glycerol, 15% (w/v) ethylene glycol, 15% dimethylsulfoxide and 0.4 M sucrose in basal medium) at room temperature, then with full strength PVS2 solution at 0°C for 50 min before direct immersion in liquid nitrogen. No regrowth was achieved after cryopreservation when shoot tips were dehydrated with PVS3 vitrification solution (50% (w/v) glycerol and 50% (w/v) sucrose in basal medium).

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“…The availability of an efficient and reliable cryogenic protocol yielding a high recovery percentage is a prerequisite for large scale, routine application of cryopreservation [10].…”

Section: Introductionmentioning

confidence: 99%

“…In these experiments, the average recovery ranged from 47% to 85%. This vitrification protocol was improved by incorporating a two-step dehydration procedure and successfully applied to ten Vitis cultivars or species, with shoot recovery ranging between 60% and 80% [10].…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation of grapevine (Vitis vinifera L.) in vitro shoot tips

Marković¹,

Chatelet

Sylvestre

et al. 2013

Open Life Sciences

Self Cite

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“…Encapsulation-dehydration experiments were performed as described by Barraco et al (2011) with the following modifications. Shoot tips were excised and then dipped into 100 or 150 mg L -1 ascorbic acid for 1 h. They were then placed in a solution of 60 g L -1 sucrose + MS for 24 h and encapsulated into calcium-alginate beads.…”

Section: Encapsulation-dehydrationmentioning

confidence: 99%

Challenges in the development of a widely applicable method for sugarcane (Saccharum spp.) shoot tip cryopreservation

Volk¹,

Jenderek²,

Staats³

et al. 2019

Acta Hortic.

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The USDA-ARS National Plant Germplasm System (NPGS) maintains 946 accessions of sugarcane (Saccharum spp.) in the field at the Subtropical Horticulture Research Station in Miami, Florida. These accessions are particularly vulnerable to hurricanes, diseases, and other threats. We sought to identify a method whereby clonally propagated sugarcane accessions could be successfully introduced into tissue culture, multiplied, and then cryopreserved as shoot tips for long-term preservation at the National Laboratory for Genetic Resources Preservation in Fort Collins, Colorado. For many accessions, 70% isopropyl alcohol and 20% commercial bleach treatments, followed by three rinses of sterile water were sufficient to remove microbial contaminants during the introduction process. However, in some cases, cefotaxime was particularly effective for removing bacterial contamination. We found that antioxidant treatments of glutathione, glycine betaine, and ascorbic acid did not improve regrowth after liquid nitrogen exposure using either PVS2 or PVS3 as cryoprotectants in droplet vitrification cryopreservation methods. Exposure durations of PVS2 and PVS3 were optimized, with and without exposure to liquid nitrogen (LN), and shoot tip regrowth levels ranged from 0 to 37% after LN exposure. Published methods for encapsulation-dehydration and V-plate vitrification cryopreservation procedures were tested to determine if acceptable results could be obtained. Using these methods, shoot tip regrowth levels ranged from 0 to 50% after LN exposure. We conclude that the sugarcane cryopreservation methods that we tested are not yet ready for implementation in the NPGS.

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“…Recently, in vitro shoot tips of two clones were successfully cryopreserved using encapsulation-dehydration according to Gonzalez-Arnao et al, (1993) and dropletvitrification with two vitrification solutions, PVS2 and PVS3 (Barraco et al, 2011). For both clones, encapsulation-dehydration induced significantly higher recovery, reaching 60% for clone H70-144 and 53% for clone CP68-1026, compared with droplet-vitrification in which recovery was 33-37% for clone H70-144 and 20-27% for clone CP68-1026.…”

Section: Apicesmentioning

confidence: 99%