Cryopreservation of Pinus radiata embryogenic tissue: effects of cryoprotective pretreatments on maturation ability

Lineros, Yenny; Balocchi, Claudio; Muñoz, Ximena; Sánchez, Manuel; Ríos, Darcy

doi:10.1007/s11240-018-1469-7

Cited by 14 publications

(10 citation statements)

References 33 publications

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“…Using Mr. Frosty container (Nalgene TM ) with a cooling rate of approximately -1 °C per min to -40 °C, recovery of tissues was on average 96.24%, with different frequencies for different cell lines (A01, A30, A31, A32). This is in line with several reports on cryopreservation of conifer embryogenic tissues using this simple and relatively cheap container (Vondrakova et al, 2010;Carneros et al, 2017;Lineros et al, 2018). Varis et al (2017) compared two slow-freezing methods using the Mr. Frosty container and programmable freezer (cooling rates 0.17 °C, 1 °C per minute).…”

Section: Relationship Between Cryotolerance and Genotypesupporting

confidence: 84%

“…The effect of the cryopreservation procedure on maturation capacity was demonstrated in several conifer species. In Pinus radiata, the maturation capacity was negatively affected by pre-treatment compared to control, but among pre-treated cell lines, no statistically differences in the number of developed mature somatic embryos were observed, albeit the genotype effect was also apparent (Lineros et al, 2018). In Abies alba, after 6 years of cryopreservation, out of 12 cell lines four cell lines showed regrowth and cotyledonary somatic embryos differentiated in two cell lines (Krajnakova et al, 2013).…”

Section: Relationship Between Cryotolerance and Genotypementioning

confidence: 99%

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Cryopreservation of Abies alba embryogenic tissues by slow-freezing method

Salaj

Panis

Klubicová

et al. 2022

Not Bot Horti Agrobo

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Embryogenic tissues of Abies alba Mill. were cryopreserved using the slow-freezing approach. Four cell lines were incubated for 24 h on a medium with 0.5 M sorbitol and pre-treated with 5% DMSO. Subsequently, the tissues were frozen at a cooling rate of 1 °C min-1 to -40 °C and transferred to liquid nitrogen for 72 hours. After thawing in a water bath at 40 °C, the tissues were cultivated on a proliferation medium. All tested lines recovered, but variations in regrowth frequencies across cell lines were noticed (91.66 to 100%). The recovered tissues showed similar features to the control 2 (non-pre-treated and non-cryopreserved tissues). In the accumulation of fresh and dry mass, no statistically significant differences were observed between cryopreserved cultures and control 2. The cryopreserved tissues produced cotyledonary somatic embryos capable of germination. Microscopic observations revealed considerable structural changes as a consequence of the cryopreservation procedure. The long vacuolated suspensor cells were disrupted, and mostly the meristematic cells of the embryonal region survived. The typical bipolar structure of early somatic embryos has been regained during the post-thaw period. Differences in cryotolerance across cell lines were also observed.

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Section: Relationship Between Cryotolerance and Genotypesupporting

confidence: 84%

Section: Relationship Between Cryotolerance and Genotypementioning

confidence: 99%

Cryopreservation of Abies alba embryogenic tissues by slow-freezing method

Salaj

Panis

Klubicová

et al. 2022

Not Bot Horti Agrobo

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“…Cryopreservation used for conifer embryogenic tissue is based on the slow-freezing method (Häggman et al, 1998; Fraga et al, 2016b; Nunes et al, 2017). Several pine species were cryopreserved for different periods of time with relatively high recovery frequencies reaching values 87% for P. pinea (Carneros et al, 2017), 100% for P. pinaster (Alvarez et al, 2012) and 100% for P. radiata (Lineros et al, 2018). In most species the maturation ability of cryopreserved and recovered tissues was restored.…”

Section: Somatic Embryogenesis In Pinus Nigra Arnmentioning

confidence: 99%

Somatic Embryogenesis in Selected Conifer Trees Pinus nigra Arn. and Abies Hybrids

et al. 2019

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Somatic embryogenesis was achieved in the conifers Pinus nigra Arn. and in the hybrids Abies alba ×A. cephalonica and Abies alba ×A. numidica. For initiation of embryogenic tissue in P. nigra, immature zygotic embryos enclosed in megagametophytes were used. The initiated embryogenic cultures were maintained and proliferated on solid culture medium DCR supplemented with 9 μM 2,4-D and 2.2 μM BA. Microscopic investigations revealed the presence of bipolar early somatic embryos in proliferating tissue. Suspension cultures have also been established by resuspending the embryogenic tissue in liquid culture medium. Experimentation with abscisic acid concentration resulted in successful somatic embryo maturation. Besides abscisic acid, the carbohydrate content or higher concentration of gelling agent in maturation medium were also important requirements for somatic embryo maturation. Germination of cotyledonary somatic embryos occurred on hormone-free medium and terminated in somatic seedlings regeneration. The regenerated somatic seedlings were transferred to soil and were capable of successful development. For initiation of embryogenic tissue in Abies hybrids juvenile explants as immature or mature zygotic embryos as well as cotyledons were used and 4.4 μM BA as sole plant growth regulator was sufficient. Medium of the same composition was also suitable for their long-term maintenance. Maturation of somatic embryos was achieved on solid DCR medium supplemented with 38 μM abscisic acid, polyethylene glycol (0, 5, 7.5, and 10% PEG-4000) and different carbohydrates such as maltose, sucrose and glucose (each 3%). PEG-4000 stimulated somatic embryo development depending on the carbohydrate source used. Cotyledonary somatic embryos germinated after desiccation treatment and the regenerated somatic seedlings were transferred to soil. Cryopreservation of embryogenic tissue could be an alternative method for long-term maintenance. For cryopreservation the slow-freezing method was used with success. Tissue regeneration in the post thaw period was relatively high and the regenerated tissue produced mature somatic embryos and subsequent plantlets. The embryogenic tissue was also used in experiments focused on genetic transformation either by biolistic (P. nigra) or Agrobacterium-mediated (Abies hybrids) methods. A proteomic study was performed to gain a deeper insight into the early stages of P. nigra somatic embryogenesis.

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“…Thus, no influence was found in Vitis spp. [ 24 ], Gentiana tibetica [ 25 ], Gentiana cruciata [ 25 , 26 ], Coriandrum sativum [ 27 ], Theobroma cacao [ 28 ], Pinus radiata [ 29 ], or Cryptomeria japonica [ 30 ]. Nevertheless, regeneration enhancement has also been described after cryopreservation.…”

Section: Discussionmentioning

confidence: 99%

Effect of Cryopreservation on the Ex Vitro Establishment of Olive Plants Regenerated via Somatic Embryogenesis

Bradaï

Sánchez-Romero

2021

Plants

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Cryopreservation is considered the best technique for the safe, long-term conservation of embryogenic cultures. However, before integrating it into a somatic embryogenesis system, the influence of cryopreservation on the final production of plants should be investigated. The objective of this investigation was to evaluate the effect of cryopreservation on the regeneration performance of olive embryogenic cultures as well as on the quality of the plants obtained and their response to ex vitro establishment. In order to analyze the influence of the genotype, all the investigations were carried out in two genetically distinct embryogenic lines. The results obtained revealed no variation in the regeneration potential or the quality of the regenerated plants due to cryopreservation. The subsequent multiplication, rooting, and acclimatization steps were not influenced by cryopreservation either, although a significant genotype × cryopreservation interaction was found for shoot length during the multiplication step. The genotype played an important role, determining the quality of the regenerated plants and some aspects of the multiplication and rooting phases. This investigation revealed that the droplet-vitrification procedure optimized for the cryopreservation of olive somatic embryos can be efficiently used for the long-term conservation of olive embryogenic lines.

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Cryopreservation of Pinus radiata embryogenic tissue: effects of cryoprotective pretreatments on maturation ability

Cited by 14 publications

References 33 publications

Cryopreservation of Abies alba embryogenic tissues by slow-freezing method

Cryopreservation of Abies alba embryogenic tissues by slow-freezing method

Somatic Embryogenesis in Selected Conifer Trees Pinus nigra Arn. and Abies Hybrids

Effect of Cryopreservation on the Ex Vitro Establishment of Olive Plants Regenerated via Somatic Embryogenesis

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