Cryopreservation of asparagus (Asparagus officinalis L.) embryogenic suspension cells and subsequent plant regeneration by vitrification

Nishizawa, Shuji; Sakai, Akira; Amano, Yoshihiko; Matsuzawa, T.

doi:10.1016/0168-9452(93)90189-7

Cited by 287 publications

(167 citation statements)

References 14 publications

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“…For each treatment, 1 mL cryoprotective loading solution was added, consisting of 2.0 M glycerol and 0.4 M sucrose (Nishizawa et al 1993). Cryovials were left for 20 min at room temperature (26 ± 2 °C) prior to the addition of a plant vitrification solution (PVS2).…”

Section: Osmoprotection and Seed Cryopreservation Treatmentsmentioning

confidence: 99%

Seed cryopreservation, in vitro propagation and ex vitro growth of Cattleya walkeriana Gardner, a vulnerable ornamental orchid

Galdiano¹,

Vendrame²,

Moretto³

et al. 2017

Aust J Crop Sci

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Section: Osmoprotection and Seed Cryopreservation Treatmentsmentioning

confidence: 99%

Seed cryopreservation, in vitro propagation and ex vitro growth of Cattleya walkeriana Gardner, a vulnerable ornamental orchid

Galdiano¹,

Vendrame²,

Moretto³

et al. 2017

Aust J Crop Sci

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“…The embryonic axes (after dehydration as described above) were submitted to vitrification in cold PVS3 solution containing 50 % (w/v) sucrose and 50 % (w/v) glycerol (Nishizawa et al 1993) or with PVS2 containing 30 % glycerol, 15 % ethylene glycol and 15 % DMSO in 0.4 M sucrose in liquid medium (Sakai et al 1990) w/v for 60 min at room temperature. Adding chilled solutions on ice may reduce the toxic effects of vitrification.…”

Section: Droplet-vitrification and Cryopreservation Of Easmentioning

confidence: 99%

Cryopreservation changes the DNA methylation of embryonic axes of Quercus robur and Fagus sylvatica seeds during in vitro culture

Nuc

Marszałek

Pukacki

2016

Trees

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Key message Dehydration and cryo-stress (2196°C) caused a greater increase in the DNA methylation of embryonic axes (EAs) of beech seeds (tolerant to both stresses) than in EAs of oak (sensitive to both stresses). Simultaneously, in vitro survival of cryopreserved beech EAs was also significantly higher than in those of oak. Abstract DNA methylation plays a key role in the regulation of growth and differentiation in plants. The present study focused on global DNA methylation of embryonic axes (EAs) isolated from the seeds of pedunculate oak (Quercus robur L.) and European beech (Fagus sylvatica L.) after cryopreservation. Oak seeds can be labeled recalcitrant, i.e., sensitive to dehydration, while beech seeds fall under the suborthodox category, i.e., tolerant to dehydration. DNA methylation was determined using a specific antibody directed at the methylated cytosine in CpG dinucleotides. The analysis was carried out on DNA isolated from EA tissues and subjected to (1) dehydration (control), (2) a combination of dehydration and plant vitrification solution (PVS3) and (3) dehydration with vitrification PVS3, and rapid cooling in liquid nitrogen (LN 2 , -196°C) at a cooling rate of 32°C s -1 . Such treatments induced global DNA methylation primarily in tolerant EAs from F. sylvatica seeds. The level of global DNA methylation in this material was 30 % higher than in the control, i.e., dehydrated (not frozen) beech EAs. The same treatment in oak EAs did not evoke such changes. After 120 days of tissue growth, we observed substantially more DNA demethylation in the beech tissues after cryopreservation in LN 2 than in the controls or in tissues subjected to vitrification but not cryopreservation. Information on the global level of DNA methylation can be useful in monitoring procedures aimed at increasing the rate of recovery of EAs following cryopreservation.

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“…0.4 M sucrose, at 22°C for 20 min and then treated with PVS2 (on ice) or PVS3 (at 22 ± 2°C) vitrification solutions for 0.5, 1, 2, and 3 h. The PVS2 consisted of 30 % (w/v) glycerol, 15 % (w/v) ethylene glycol, 15 % (w/v) DMSO and 0.4 M sucrose in 1/2MS culture medium (Sakai et al 1990). The PVS3 solution contained 40 % (w/v) glycerol and 40 % (w/v) sucrose (Nishizawa et al 1993). Samples were washed twice in fresh PVS2 or PVS3 solutions, loaded into 2-ml cryovials and placed directly into LN for 24 h. For rewarming, the cryotubes were plunged into a water bath at 35°C for 1.5 min; the gametophytes were then transferred to 1.2 M sucrose solution for 0.5 h. The explants were subsequently placed on 1/2MS agar medium in darkness for 2 days.…”

Section: Gametophyte Cryopreservation and Survival Assessmentmentioning

confidence: 99%

Integration of tissue culture and cryopreservation methods for propagation and conservation of the fern Osmunda regalis L.

et al. 2015

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A simple and reliable technique for in vitro multiplication and long-term preservation using liquid nitrogen was developed for gametophytes of Osmunda regalis. The effect of Knop's medium and various concentrations (1/2, 1/4, 1/8) of mineral salts provided in Murashige and Skoog (MS) basal medium, together in the presence or absence of both ammonium nitrate and a full complement of vitamins on gametophyte proliferation and sporophyte production was determined. Moreover, the effectiveness of gametophyte cryopreservation by vitrification and encapsulation-vitrification techniques was assessed. Maximum gametophyte proliferation (89 %) occurred on the ammonium nitrate-and vitamin-free [(-NH4NO3)(-vit)] MS medium with 1/2 or 1/4 strength mineral salts. The maximum production of sporophytes (30 plantlets per gametophyte clump) required 1/8MS (-NH4NO3)(-vit) medium. The flow cytometric analysis revealed that the sporophytes contained twofold more pg DNA than gametophytes. This confirmed that the sporophytes were obtained by sexual reproduction. The vitrification protocol and PVS2 solution were ineffective for cryopreservation. The greatest survival rate (81.6 %) following cryo-exposure occurred following treatment of encapsulated gametophytes with PVS3 solution for 3 h. This protocol allowed the recovery of gametophyte cultures following 6 weeks after rewarming. Finally, 100 % of sporophytes produced in vitro were successfully acclimated to ex vitro conditions. Application of the in vitro and cryopreservation methods made it possible to improve the number and time of O. regalis sporophyte production. Whole system of micropropagation can be completed in approximately 1 year. The protocols open new avenues for the mass propagation, germplasm conservation and resource management of the species.

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Cryopreservation of asparagus (Asparagus officinalis L.) embryogenic suspension cells and subsequent plant regeneration by vitrification

Cited by 287 publications

References 14 publications

Seed cryopreservation, in vitro propagation and ex vitro growth of Cattleya walkeriana Gardner, a vulnerable ornamental orchid

Seed cryopreservation, in vitro propagation and ex vitro growth of Cattleya walkeriana Gardner, a vulnerable ornamental orchid

Cryopreservation changes the DNA methylation of embryonic axes of Quercus robur and Fagus sylvatica seeds during in vitro culture

Integration of tissue culture and cryopreservation methods for propagation and conservation of the fern Osmunda regalis L.

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