Droplet-vitrification of apical shoot tips of Rubus fruticosus L. and Prunus cerasifera Ehrh.
Abstract
Apical shoot tips excised from in vitro plantlets of blackberry (Rubus fruticosus L. 'Cacanska Bestrna') and cherry plum (Prunus cerasifera Ehrh.) were tested for recovery after cryopreservation using the droplet-vitrification technique. Following treatment for 30 min with a loading solution comprising 1.9 M glycerol and 0.5 M sucrose, explants were dehydrated with a highly concentrated cryoprotectant solution, so called vitrification solution. Shoot tips were dehydrated for 10,20 and 30 min at room temperature with a solution derived from the original PVS2 solution (containing 37.5% (w/v) glycerol, 15% (w/v) dimethylsulfoxide, 15% (w/v) ethylene glycol and 22.5% (w/v) sucrose) and for 60,90 and 120 min using the PVS3 solution (containing 50% (w/v) glycerol and 50% (w/v) sucrose). Explants were cooled by direct immersion in LN in 10 mu l droplets of vitrification solution placed on aluminium foil strips. Rewarming was done by direct plunging of foil strips in a preheated (37 degrees C)... unloading solution (0.8 M sucrose) for 30 s, after which an equal volume of unloading solution (at room temperature) was added for further incubation for 30 min. As for regrowth of blackberry, PVS3 proved more effective than the modified PVS2, but the difference was significant (P LT 0.05) only for the shortest treatment duration. The duration of PVS3 treatment had no significant effect on regrowth of cryopreserved shoot tips (45.8-70%). By contrast, a 30-min treatment with modified PVS2 solution resulted in a significant increase in regeneration percentage (30%), as compared with a 10-min treatment with the same solution (5%). Cherry plum shoot tips were very sensitive to both vitrification solutions and growth recovery of cryopreserved samples was generally lower (5-20%) than that of blackberry explants. No significant influence of PVS treatment (both type of solution and treatment duration) on regrowth of cryopreserved shoot tips was observed with cherry plum shoot tips. Experiments performed in France and in Serbia produced similar results, thereby showing the robustness and reproducibility of the protocols developed.
Keywords:
Vitrification solution / In vitro shoot tips / Dehydration / Cryopreservation / Cherry plum / BlackberrySource:
Scientia Horticulturae, 2011, 130, 1, 222-228Publisher:
- Elsevier, Amsterdam
Funding / projects:
- COST Action [STSM 871-06145]
- Development and preservation of genetic potential of temperate zone fruits (RS-31064)
- ARCAD
- Agropolis Foundation (Montpellier, France)
DOI: 10.1016/j.scienta.2011.06.049
ISSN: 0304-4238
WoS: 000294942900035
Scopus: 2-s2.0-79961173350
Collections
Institution/Community
Institut za voćarstvoTY - JOUR AU - Vujović, Tatjana AU - Sylvestre, Isabelle AU - Ružić, Đurđina AU - Engelmann, Florent PY - 2011 UR - https://refri.institut-cacak.org/handle/123456789/209 AB - Apical shoot tips excised from in vitro plantlets of blackberry (Rubus fruticosus L. 'Cacanska Bestrna') and cherry plum (Prunus cerasifera Ehrh.) were tested for recovery after cryopreservation using the droplet-vitrification technique. Following treatment for 30 min with a loading solution comprising 1.9 M glycerol and 0.5 M sucrose, explants were dehydrated with a highly concentrated cryoprotectant solution, so called vitrification solution. Shoot tips were dehydrated for 10,20 and 30 min at room temperature with a solution derived from the original PVS2 solution (containing 37.5% (w/v) glycerol, 15% (w/v) dimethylsulfoxide, 15% (w/v) ethylene glycol and 22.5% (w/v) sucrose) and for 60,90 and 120 min using the PVS3 solution (containing 50% (w/v) glycerol and 50% (w/v) sucrose). Explants were cooled by direct immersion in LN in 10 mu l droplets of vitrification solution placed on aluminium foil strips. Rewarming was done by direct plunging of foil strips in a preheated (37 degrees C) unloading solution (0.8 M sucrose) for 30 s, after which an equal volume of unloading solution (at room temperature) was added for further incubation for 30 min. As for regrowth of blackberry, PVS3 proved more effective than the modified PVS2, but the difference was significant (P LT 0.05) only for the shortest treatment duration. The duration of PVS3 treatment had no significant effect on regrowth of cryopreserved shoot tips (45.8-70%). By contrast, a 30-min treatment with modified PVS2 solution resulted in a significant increase in regeneration percentage (30%), as compared with a 10-min treatment with the same solution (5%). Cherry plum shoot tips were very sensitive to both vitrification solutions and growth recovery of cryopreserved samples was generally lower (5-20%) than that of blackberry explants. No significant influence of PVS treatment (both type of solution and treatment duration) on regrowth of cryopreserved shoot tips was observed with cherry plum shoot tips. Experiments performed in France and in Serbia produced similar results, thereby showing the robustness and reproducibility of the protocols developed. PB - Elsevier, Amsterdam T2 - Scientia Horticulturae T1 - Droplet-vitrification of apical shoot tips of Rubus fruticosus L. and Prunus cerasifera Ehrh. EP - 228 IS - 1 SP - 222 VL - 130 DO - 10.1016/j.scienta.2011.06.049 UR - conv_1402 ER -
@article{ author = "Vujović, Tatjana and Sylvestre, Isabelle and Ružić, Đurđina and Engelmann, Florent", year = "2011", abstract = "Apical shoot tips excised from in vitro plantlets of blackberry (Rubus fruticosus L. 'Cacanska Bestrna') and cherry plum (Prunus cerasifera Ehrh.) were tested for recovery after cryopreservation using the droplet-vitrification technique. Following treatment for 30 min with a loading solution comprising 1.9 M glycerol and 0.5 M sucrose, explants were dehydrated with a highly concentrated cryoprotectant solution, so called vitrification solution. Shoot tips were dehydrated for 10,20 and 30 min at room temperature with a solution derived from the original PVS2 solution (containing 37.5% (w/v) glycerol, 15% (w/v) dimethylsulfoxide, 15% (w/v) ethylene glycol and 22.5% (w/v) sucrose) and for 60,90 and 120 min using the PVS3 solution (containing 50% (w/v) glycerol and 50% (w/v) sucrose). Explants were cooled by direct immersion in LN in 10 mu l droplets of vitrification solution placed on aluminium foil strips. Rewarming was done by direct plunging of foil strips in a preheated (37 degrees C) unloading solution (0.8 M sucrose) for 30 s, after which an equal volume of unloading solution (at room temperature) was added for further incubation for 30 min. As for regrowth of blackberry, PVS3 proved more effective than the modified PVS2, but the difference was significant (P LT 0.05) only for the shortest treatment duration. The duration of PVS3 treatment had no significant effect on regrowth of cryopreserved shoot tips (45.8-70%). By contrast, a 30-min treatment with modified PVS2 solution resulted in a significant increase in regeneration percentage (30%), as compared with a 10-min treatment with the same solution (5%). Cherry plum shoot tips were very sensitive to both vitrification solutions and growth recovery of cryopreserved samples was generally lower (5-20%) than that of blackberry explants. No significant influence of PVS treatment (both type of solution and treatment duration) on regrowth of cryopreserved shoot tips was observed with cherry plum shoot tips. Experiments performed in France and in Serbia produced similar results, thereby showing the robustness and reproducibility of the protocols developed.", publisher = "Elsevier, Amsterdam", journal = "Scientia Horticulturae", title = "Droplet-vitrification of apical shoot tips of Rubus fruticosus L. and Prunus cerasifera Ehrh.", pages = "228-222", number = "1", volume = "130", doi = "10.1016/j.scienta.2011.06.049", url = "conv_1402" }
Vujović, T., Sylvestre, I., Ružić, Đ.,& Engelmann, F.. (2011). Droplet-vitrification of apical shoot tips of Rubus fruticosus L. and Prunus cerasifera Ehrh.. in Scientia Horticulturae Elsevier, Amsterdam., 130(1), 222-228. https://doi.org/10.1016/j.scienta.2011.06.049 conv_1402
Vujović T, Sylvestre I, Ružić Đ, Engelmann F. Droplet-vitrification of apical shoot tips of Rubus fruticosus L. and Prunus cerasifera Ehrh.. in Scientia Horticulturae. 2011;130(1):222-228. doi:10.1016/j.scienta.2011.06.049 conv_1402 .
Vujović, Tatjana, Sylvestre, Isabelle, Ružić, Đurđina, Engelmann, Florent, "Droplet-vitrification of apical shoot tips of Rubus fruticosus L. and Prunus cerasifera Ehrh." in Scientia Horticulturae, 130, no. 1 (2011):222-228, https://doi.org/10.1016/j.scienta.2011.06.049 ., conv_1402 .