Cryopreservation of Viola cornuta shoot tips using vitrification procedure
Аутори
Trajković, MilenaAntonić, Dragana
Trailović, Maja
Trifunović Momčilov, Milana
Subotić, Angelina
Jevremović, Slađana
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Cryopreservation represents a suitable method for long term storage of different plant genetic
resources. The aim of this study was to develop protocol for cryopreservation of Viola cornuta
shoot tips using one step freezing method with chemical dehydration of tissue with modified
Plant Vitrification Solutions (PVS2 or PVS3). Shoot tips (1-2 mm) of two-week cold acclimated
shoots were cultured on ½MS medium with 0.3 M sucrose for one day before treatment with loading solution (2 M glycerol, 0.4 M sucrose) for 30 min. Osmotic dehydration with PVS2 solution (30% glycerol, 15% ethylene glycol and 15% DMSO in liquid ½MS medium with 0.4 M sucrose) were tested at 0 °C or 24 °C. Osmotic dehydration with PVS3 (50% sucrose, 50% glycerol in liquid ½MS medium) were tested at 24 °C for 45 min. After the treatment the explants were directly immersed in liquid nitrogen (LN) for at least one day. Re-warming was performed at 42 °C in water bath for 2 min. After re-warming, the PVS solutions were r...eplaced with unloading solution containing 1.2 M sucrose for 20 min. Re-warmed shoot tips were cultured on ½MS medium with 0.1 mg L-1 BAP. We observed that PVS2 solution is cytotoxic for V. cornuta shoot tips and cannot be used for cryopreservation. However, cryopreservation with PVS3 solution was successful, where 71.9-100% shoot tips survived treatment before immersion to LN and 31-40% survived after re-warming from LN. Regrowth of cryopreserved shoot tips with new well-formed leaves was obtained after four weeks of culture.
Кључне речи:
horned pancy / Plant Vitrification Solution / PVS2 / PVS3Извор:
3rd International Conference on Plant Biology (22nd SPPS Meeting) 9-12 June 2018, Belgrade, 2018Издавач:
- Serbian Plant Physiology Society Institute for Biological Research “Siniša Stanković”, University of Belgrade Faculty of Biology, University of Belgrade
Институција/група
Institut za multidisciplinarna istraživanjaTY - CONF AU - Trajković, Milena AU - Antonić, Dragana AU - Trailović, Maja AU - Trifunović Momčilov, Milana AU - Subotić, Angelina AU - Jevremović, Slađana PY - 2018 UR - http://rimsi.imsi.bg.ac.rs/handle/123456789/2365 AB - Cryopreservation represents a suitable method for long term storage of different plant genetic resources. The aim of this study was to develop protocol for cryopreservation of Viola cornuta shoot tips using one step freezing method with chemical dehydration of tissue with modified Plant Vitrification Solutions (PVS2 or PVS3). Shoot tips (1-2 mm) of two-week cold acclimated shoots were cultured on ½MS medium with 0.3 M sucrose for one day before treatment with loading solution (2 M glycerol, 0.4 M sucrose) for 30 min. Osmotic dehydration with PVS2 solution (30% glycerol, 15% ethylene glycol and 15% DMSO in liquid ½MS medium with 0.4 M sucrose) were tested at 0 °C or 24 °C. Osmotic dehydration with PVS3 (50% sucrose, 50% glycerol in liquid ½MS medium) were tested at 24 °C for 45 min. After the treatment the explants were directly immersed in liquid nitrogen (LN) for at least one day. Re-warming was performed at 42 °C in water bath for 2 min. After re-warming, the PVS solutions were replaced with unloading solution containing 1.2 M sucrose for 20 min. Re-warmed shoot tips were cultured on ½MS medium with 0.1 mg L-1 BAP. We observed that PVS2 solution is cytotoxic for V. cornuta shoot tips and cannot be used for cryopreservation. However, cryopreservation with PVS3 solution was successful, where 71.9-100% shoot tips survived treatment before immersion to LN and 31-40% survived after re-warming from LN. Regrowth of cryopreserved shoot tips with new well-formed leaves was obtained after four weeks of culture. PB - Serbian Plant Physiology Society Institute for Biological Research “Siniša Stanković”, University of Belgrade Faculty of Biology, University of Belgrade C3 - 3rd International Conference on Plant Biology (22nd SPPS Meeting) 9-12 June 2018, Belgrade T1 - Cryopreservation of Viola cornuta shoot tips using vitrification procedure UR - https://hdl.handle.net/21.15107/rcub_rimsi_2365 ER -
@conference{ author = "Trajković, Milena and Antonić, Dragana and Trailović, Maja and Trifunović Momčilov, Milana and Subotić, Angelina and Jevremović, Slađana", year = "2018", abstract = "Cryopreservation represents a suitable method for long term storage of different plant genetic resources. The aim of this study was to develop protocol for cryopreservation of Viola cornuta shoot tips using one step freezing method with chemical dehydration of tissue with modified Plant Vitrification Solutions (PVS2 or PVS3). Shoot tips (1-2 mm) of two-week cold acclimated shoots were cultured on ½MS medium with 0.3 M sucrose for one day before treatment with loading solution (2 M glycerol, 0.4 M sucrose) for 30 min. Osmotic dehydration with PVS2 solution (30% glycerol, 15% ethylene glycol and 15% DMSO in liquid ½MS medium with 0.4 M sucrose) were tested at 0 °C or 24 °C. Osmotic dehydration with PVS3 (50% sucrose, 50% glycerol in liquid ½MS medium) were tested at 24 °C for 45 min. After the treatment the explants were directly immersed in liquid nitrogen (LN) for at least one day. Re-warming was performed at 42 °C in water bath for 2 min. After re-warming, the PVS solutions were replaced with unloading solution containing 1.2 M sucrose for 20 min. Re-warmed shoot tips were cultured on ½MS medium with 0.1 mg L-1 BAP. We observed that PVS2 solution is cytotoxic for V. cornuta shoot tips and cannot be used for cryopreservation. However, cryopreservation with PVS3 solution was successful, where 71.9-100% shoot tips survived treatment before immersion to LN and 31-40% survived after re-warming from LN. Regrowth of cryopreserved shoot tips with new well-formed leaves was obtained after four weeks of culture.", publisher = "Serbian Plant Physiology Society Institute for Biological Research “Siniša Stanković”, University of Belgrade Faculty of Biology, University of Belgrade", journal = "3rd International Conference on Plant Biology (22nd SPPS Meeting) 9-12 June 2018, Belgrade", title = "Cryopreservation of Viola cornuta shoot tips using vitrification procedure", url = "https://hdl.handle.net/21.15107/rcub_rimsi_2365" }
Trajković, M., Antonić, D., Trailović, M., Trifunović Momčilov, M., Subotić, A.,& Jevremović, S.. (2018). Cryopreservation of Viola cornuta shoot tips using vitrification procedure. in 3rd International Conference on Plant Biology (22nd SPPS Meeting) 9-12 June 2018, Belgrade Serbian Plant Physiology Society Institute for Biological Research “Siniša Stanković”, University of Belgrade Faculty of Biology, University of Belgrade.. https://hdl.handle.net/21.15107/rcub_rimsi_2365
Trajković M, Antonić D, Trailović M, Trifunović Momčilov M, Subotić A, Jevremović S. Cryopreservation of Viola cornuta shoot tips using vitrification procedure. in 3rd International Conference on Plant Biology (22nd SPPS Meeting) 9-12 June 2018, Belgrade. 2018;. https://hdl.handle.net/21.15107/rcub_rimsi_2365 .
Trajković, Milena, Antonić, Dragana, Trailović, Maja, Trifunović Momčilov, Milana, Subotić, Angelina, Jevremović, Slađana, "Cryopreservation of Viola cornuta shoot tips using vitrification procedure" in 3rd International Conference on Plant Biology (22nd SPPS Meeting) 9-12 June 2018, Belgrade (2018), https://hdl.handle.net/21.15107/rcub_rimsi_2365 .