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

ABSTRACT: 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.

SUBMITTER: Bradai F

PROVIDER: S-EPMC7921972 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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

Bradaï Fatiha F Sánchez-Romero Carolina C

Plants (Basel, Switzerland) 20210219 2

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 est ...[more]

PMID: 33669606

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Project description:The effects of growth regulators, namely, 6-benzylaminopurine (BAP) and thidiazuron (TDZ), on the morphogenic capacity of 13 cultivars of clematis plants, in terms of their morphological structure formation, shoot regeneration, and somatic embryo development, are presented. The clematis cultivars 'Alpinist,' 'Ay-Nor,' 'Bal Tsvetov,' 'Crimson Star,' 'Crystal Fountain,' 'Kosmicheskaya Melodiya,' 'Lesnaya Opera,' 'Madame Julia Correvon,' 'Nevesta,' 'Nikitsky Rosovyi,' 'Nikolay Rubtsov,' 'Serenada Kryma,' and 'Vechniy Zov' were taken in collection plots of the Nikita Botanical Gardens for use in study. After explant sterilization with 70% ethanol (1 min), 0.3-0.4% Cl2 (15 min), and 1% thimerosal (10 min), 1-cm long segments with a single node were introduced to an in vitro culture. The explants were established on the basal MS medium supplemented with BAP (2.20-8.90 μM) and 0.049 μM NAA, or TDZ (3.0; 6.0, and 9.0 μM) with 30 g/L sucrose and 9 g/L agar. The medium with 0.89 μM BAP served as the control. Culture vessels and test tubes with the explants were maintained in plant growth chamber-controlled conditions: with a 16-h photoperiod, under cool-white light fluorescent lamps with a light intensity of 37.5 μmol m-2 s-1, at a temperature of 24 ± 1°C. Histological analysis demonstrated that adventitious bud and somatic embryo formation in studied clematis cultivars occurred at numerous areas of active meristematic cell zones. The main role of plant growth regulators and its concentrations were demonstrated. It was determined that maximum adventitious microshoot regeneration without any morphological abnormalities formed on the media supplemented with BAP or TDZ. 4.40 μM BAP, or 6.0 μM TDZ were optimal cytokinin concentrations for micropropagation. The explants of 'Alpinist,' 'Ay-Nor,' 'Crimson Star,' 'Crystal Fountain,' 'Nevesta,' and 'Serenada Kryma' cultivars displayed high morphogenetic capacity under in vitro culturing. During indirect somatic embryogenesis, light intensity 37.5 μmol m-2 s-1 stimulated a higher-number somatic embryo formation and a temperature of 26°C affected somatic embryo development. Active formation of primary and secondary somatic embryos was also demonstrated. 2.20 μM BAP with 0.09 μM IBA affected the high-number somatic embryo formation for eight cultivars. Secondary somatic embryogenesis by the same concentration of BAP was induced. The frequency of secondary somatic embryogenesis was higher in 'Crystal Fountain' (100%), 'Crimson Star' (100%), 'Nevesta' (97%), and 'Ay-Nor' (92%) cultivars. Based on these results, the methodology for direct somatic embryogenesis and organogenesis of studied clematis cultivars has been developed.

Dataset Information

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

Publications

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

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