Cryopreservation of Abies alba embryogenic tissues by slow-freezing method
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 liqui...
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| Format: | Article |
| Language: | English |
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AcademicPres
2022-12-01
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| Series: | Notulae Botanicae Horti Agrobotanici Cluj-Napoca |
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| Online Access: | https://www.notulaebotanicae.ro/index.php/nbha/article/view/12770 |
| _version_ | 1811198543185051648 |
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| author | Terezia SALAJ Bart PANIS Katarina KLUBICOVA Jan SALAJ |
| author_facet | Terezia SALAJ Bart PANIS Katarina KLUBICOVA Jan SALAJ |
| author_sort | Terezia SALAJ |
| collection | DOAJ |
| description |
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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| first_indexed | 2024-04-12T01:33:37Z |
| format | Article |
| id | doaj.art-f7f121fbe5c54da7aee78fdd46d76816 |
| institution | Directory Open Access Journal |
| issn | 0255-965X 1842-4309 |
| language | English |
| last_indexed | 2024-04-12T01:33:37Z |
| publishDate | 2022-12-01 |
| publisher | AcademicPres |
| record_format | Article |
| series | Notulae Botanicae Horti Agrobotanici Cluj-Napoca |
| spelling | doaj.art-f7f121fbe5c54da7aee78fdd46d768162022-12-22T03:53:25ZengAcademicPresNotulae Botanicae Horti Agrobotanici Cluj-Napoca0255-965X1842-43092022-12-0150410.15835/nbha50312770Cryopreservation of Abies alba embryogenic tissues by slow-freezing methodTerezia SALAJ0Bart PANIS1Katarina KLUBICOVA2Jan SALAJ3Plant Science and Biodiversity Center SAS, Institute of Plant Genetics and Biotechnology, Akademicka 2, 950 07 NitraAlliance of Biodiversity International and International and CIAT, c/o KU Leuven, Willem de Croylaan 42, 3001 Leuven; Department of Biosystems, KU Leuven, Willem de Croylaan 42, 3001 LeuvenPlant Science and Biodiversity Center SAS, Institute of Plant Genetics and Biotechnology, Akademicka 2, 950 07 NitraPlant Science and Biodiversity Center SAS, Institute of Plant Genetics and Biotechnology, Akademicka 2, 950 07 Nitra 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. https://www.notulaebotanicae.ro/index.php/nbha/article/view/12770coniferscryo-toleranceregenerationsilver firsomatic embryogenesis |
| spellingShingle | Terezia SALAJ Bart PANIS Katarina KLUBICOVA Jan SALAJ Cryopreservation of Abies alba embryogenic tissues by slow-freezing method Notulae Botanicae Horti Agrobotanici Cluj-Napoca conifers cryo-tolerance regeneration silver fir somatic embryogenesis |
| title | Cryopreservation of Abies alba embryogenic tissues by slow-freezing method |
| title_full | Cryopreservation of Abies alba embryogenic tissues by slow-freezing method |
| title_fullStr | Cryopreservation of Abies alba embryogenic tissues by slow-freezing method |
| title_full_unstemmed | Cryopreservation of Abies alba embryogenic tissues by slow-freezing method |
| title_short | Cryopreservation of Abies alba embryogenic tissues by slow-freezing method |
| title_sort | cryopreservation of abies alba embryogenic tissues by slow freezing method |
| topic | conifers cryo-tolerance regeneration silver fir somatic embryogenesis |
| url | https://www.notulaebotanicae.ro/index.php/nbha/article/view/12770 |
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