Effect of sucrose preculture and culture inoculum density on cryopreservation of olive somatic embryos

Bradaï, Fatiha; Almagro-Bastante, Javier; Sánchez-Romero, Carolina

doi:10.1016/j.scienta.2023.112385

Cited by 4 publications

(2 citation statements)

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“…Culture conditions of somatic embryos had a determinant influence on their subsequent response to cryopreservation. Preculture in liquid medium supplemented with 0.2 M sucrose for 28 days significantly improved cryopreservation, with 90% of explants resuming embryogenesis after rewarming ( Bradaï et al., 2023 ).…”

Section: Cryopreservation Of Embryogenic Materials In Woody Speciesmentioning

confidence: 99%

“…Once an embryogenic line has been established, periodic subculture is necessary to maintain the embryogenic ability. This task is labor-intensive and there are also a risks of losing lines by contamination, technical failure, human mistakes, reduction of the embryogenic ability, or the appearance of somaclonal variation ( Bradaï et al., 2023 ). The development and optimization of efficient cryopreservation procedures enable safe and long-term conservation of embryogenic cultures ( Ozudogru and Lambardi, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

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Current status of the cryopreservation of embryogenic material of woody species

Ballesteros,

Martínez,

Sánchez-Romero

et al. 2024

Front. Plant Sci.

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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.

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Section: Cryopreservation Of Embryogenic Materials In Woody Speciesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Current status of the cryopreservation of embryogenic material of woody species

Ballesteros,

Martínez,

Sánchez-Romero

et al. 2024

Front. Plant Sci.

Self Cite

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Cryopreservation of embryogenic callus in Oryza sativa L.: Assessment of impact of callus age on regeneration; morphological and genetic stability regenerants

Zeliang,

Pattanayak

2024

Plant Cell Tiss Organ Cult

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Cryopreservation of sessile oak (Quercus petraea (Matt.) Liebl.) plumules using aluminium cryo-plates: influence of cryoprotection and drying

Wasileńczyk,

Wawrzyniak,

Martins

et al. 2024

Plant Methods

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Background Quercus seeds that are recalcitrant to desiccation and freezing temperatures cannot be stored in gene banks under conventional conditions. However, the germplasm of some recalcitrant seeded species can be stored in liquid nitrogen (–196 °C). Unfortunately, for many species, among them for almost the whole genus Quercus, an effective cryostorage method is still unknown. In this study, we propose a successful cryostorage protocol for Quercus petraea (Matt.) Liebl. germplasm using plumules (a shoot apical meristem of an embryo) frozen on aluminium cryo-plates. Results The plumules isolated from the acorns of ten provenances were prestored in 0.5 M sucrose solution (for 18 h). To form alginate beads (one plumule per bead), the plumules were placed in the wells of a cryo-plate and embedded in calcium alginate gel. For cryoprotection, the encapsulated plumules were immersed in cryoprotectant solution containing 2.0 M glycerol and different concentrations of sucrose (0.8–1.2 M) for 40 min at 25 °C and desiccated under a laminar flow cabinet for 1.0–4.0 h. Cryo-plates with plumules were directly immersed in liquid nitrogen and then cryostored for 30 min. For rewarming, cryo-plates with plumules were immersed in 1.0 M sucrose solution and rehydrated for 15 min at 25 °C. Survival rates varied from 25.8 to 83.4 were achieved after cryoprotection in 1.0 M sucrose solution and the drying of plumules for 2 h. The in vitro regrowth rate of cryopreserved plumules varied among provenances and was 26–77%. Conclusions This study presents, for the first time, a successful, simple and effective protocol for the cryopreservation of Q. petraea germplasm that could be used in gene banks. The experiment was successfully repeated on seeds from various provenances, each yielding similar, good results. However, seed quality and storage time after harvesting are important factors in plumule regrowth after cryopreservation.

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Effect of sucrose preculture and culture inoculum density on cryopreservation of olive somatic embryos

Cited by 4 publications

References 48 publications

Current status of the cryopreservation of embryogenic material of woody species

Current status of the cryopreservation of embryogenic material of woody species

Cryopreservation of embryogenic callus in Oryza sativa L.: Assessment of impact of callus age on regeneration; morphological and genetic stability regenerants

Cryopreservation of sessile oak (Quercus petraea (Matt.) Liebl.) plumules using aluminium cryo-plates: influence of cryoprotection and drying

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