Cryopreservation of olive somatic embryos using the droplet-vitrification method: The importance of explant culture conditions

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

doi:10.1016/j.scienta.2017.02.003

Cited by 14 publications

(16 citation statements)

References 33 publications

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“…Some studies on cryopreservation of SEs of avocado [ 54 ] and olive [ 55 ] have reported 30 min of dehydration as the optimal exposure time to PVS2 using a droplet-vitrification procedure. There are other cases in which more prolonged exposures (60–90 min) have proved to be beneficial [ 26 , 28 , 53 ].…”

Section: Discussionmentioning

confidence: 99%

Indirect Somatic Embryogenesis and Cryopreservation of Agave tequilana Weber Cultivar ‘Chato’

Delgado-Aceves

González-Arnao

Santacruz-Ruvalcaba

et al. 2021

Plants

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Agave tequilana Weber cultivar ‘Chato’ represents an important genetic supply of wild severely in decline populations of ‘Chato’ for breeding and transformation programs. In this work, the indirect somatic embryogenesis and cryopreservation of Somatic Embryos (SEs) were investigated using the ‘Chato’ cultivar as a study case. Methods: Embryogenic calli were induced by the cultivation of 1 cm of young leaves from in vitro plants on MS semisolid medium supplemented with 24.84, 33.13, 41.41, 49.69, and 57.98 μM 4-amino-3,5,6-trichloro-2- pyridinecarboxylic acid (picloram) in combination with 2.21, 3.32, and 4.43 μM 6-benzylaminopurine (BAP). The origin and structure of formed SEs were verified by histological analysis. Cryopreservation studies of SEs were performed following the V-cryoplate technique and using for dehydration two vitrification solutions (PVS2 and PVS3). Results: The highest average (52.43 ± 5.74) of produced SEs and the Embryo Forming Capacity (estimated index 52.43) were obtained using 49.69 µM picloram and 3.32 µM BAP in the culture medium. The highest post-cryopreservation regrowth (83%) and plant conversion rate (around 70%) were achieved with PVS2 at 0 °C for 15 min. Conclusion: Our work provides new advances about somatic embryogenesis in Agave and reports the first results on cryopreservation of SEs of this species.

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Section: Discussionmentioning

confidence: 99%

Indirect Somatic Embryogenesis and Cryopreservation of Agave tequilana Weber Cultivar ‘Chato’

Delgado-Aceves

González-Arnao

Santacruz-Ruvalcaba

et al. 2021

Plants

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“…Five to six months after rewarming, cultures established from somatic embryos cryopreserved following the protocol of Bradaï et al [ 13 ] presented a good aspect. Although in general, higher proliferation rates were found in cryopreserved lines compared to control, non-frozen cultures; as found in previous investigations [ 14 , 15 ], the genotype exerted a primary effect, determining the significance of the influence.…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, embryogenic cultures established after rewarming retain their morphogenic capacity for subsequent regeneration of plants, with no evident negative effects affecting the somatic embryogenesis process executed following the procedure described by Sánchez-Romero [ 6 ]. Therefore, the droplet-vitrification method optimized by Bradaï et al [ 13 ] can be considered a reliable cryopreservation procedure for the long-term conservation of olive embryogenic lines.…”

Section: Discussionmentioning

confidence: 99%

“…Cryopreservation of olive somatic embryos was carried out following the protocol of Bradaï et al [ 13 ], using the droplet vitrification method on aluminum foil strips [ 46 ]. Briefly, somatic embryos 1–6 mm in size were selected from stock cultures of two independent embryogenic lines (T1 and P5) at the end of a maintenance cycle.…”

Section: Methodsmentioning

confidence: 99%

“…Both classical, based on slow cooling protocols, and new, vitrification-based methods have been used to conserve olive embryogenic cultures [ 12 ]. In 2017, Bradaï et al [ 13 ] achieved successful cryopreservation of olive somatic embryos by using the droplet-vitrification method on aluminum foil strips. Using this procedure, 60% of samples resumed embryogenesis, thus ensuring the safe long-term conservation of this type of embryogenic structures.…”

Section: Introductionmentioning

confidence: 99%

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Effect of Cryopreservation on Olive (Olea europaea L.) Plant Regeneration via Somatic Embryogenesis

Bradaï

Sánchez-Romero

2020

Plants

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Olive somatic embryos have been successfully cryopreserved using the droplet-vitrification method on aluminum foil strips. Although acceptable recovery rates have been obtained after rewarming, the influence of this cryopreservation protocol on the somatic embryogenesis process is unknown. To evaluate the effect of cryopreservation on olive somatic embryogenesis, the behavior of cultures established from cryopreserved somatic embryos was compared with that of control, non-cryopreserved cultures in the different phases of the somatic embryogenesis process. In order to analyze the influence of the genotype, this investigation was carried out in two independent lines. During the proliferation step, only the line T1 was affected by cryopreservation, with higher fresh weight increases. Although similar total embryos were produced per culture, freezing in liquid nitrogen significantly improved the maturation pattern in the line P5. Better germination results were also found in this embryogenic line. The genotype plays a key role, largely determining the effect of cryopreservation on olive somatic embryogenesis. A specific genotype-dependent response was found depending on the culture step. Variations observed could not be associated to differences in the embryogenic lines’ instability to maintain their morphogenic competence after cryopreservation. Embryogenic cultures established after rewarming retained their regeneration capacity, with no evident negative effects affecting their regeneration capacity.

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Synthetic Seeds: Prospects and Advances in Cryopreservation

2019

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Cryopreservation of olive somatic embryos using the droplet-vitrification method: The importance of explant culture conditions

Cited by 14 publications

References 33 publications

Indirect Somatic Embryogenesis and Cryopreservation of Agave tequilana Weber Cultivar ‘Chato’

Indirect Somatic Embryogenesis and Cryopreservation of Agave tequilana Weber Cultivar ‘Chato’

Effect of Cryopreservation on Olive (Olea europaea L.) Plant Regeneration via Somatic Embryogenesis

Synthetic Seeds: Prospects and Advances in Cryopreservation

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