Plant regeneration from Rosa shoot tips cryopreserved by a combined droplet vitrification method |
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Authors: | Adela Halmagyi Ina Pinker |
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Affiliation: | (1) Institute of Biological Research, Republicii str. 48, 400015 Cluj-Napoca, Romania;(2) Institute of Horticultural Sciences, Humboldt University Berlin, Albrecht-Thaer Weg 1, 14195 Berlin, Germany |
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Abstract: | Shoot tips obtained from in vitro Rosa plants (three cultivars) were successfully cryopreserved by a combined droplet vitrification method and subsequently shoots regenerated. The excised shoot tips (1–4 mm long) were incubated in a liquid MS medium supplemented with 2.5 mg l−1 thiamine, 0.2 mg l−1 biotin, 0.2 mg l−1 pyridoxine, 0.25 mg l−1 6-benzylaminopurine (BAP), 0.5 mg l−1 gibberellic acid (GA3) and 0.08 M sucrose, for 24 h. Following that incubation shoot tips were pre-cultured in this MS medium containing 0.1 till 1.0 M sucrose for 24 and 48 h, respectively. Pre-cultured shoot tips were dehydrated with concentrated PVS2 cryoprotective solution for 10–30 min at room temperature, prior to a direct plunge in liquid nitrogen. After rapid rewarming in the above mentioned liquid medium shoot tips were plated on a modified MS medium (5 g l−1 agar) supplemented with vitamins and plant growth regulators as mentioned above for regrowth. Cryopreserved shoot tips resumed growth within 10 days and regenerated shoots within 3 weeks. The highest numbers of regrowing shoot tips were 64.44% for cv. Kardinal, 67.73% for cv. Fairy and 57.57% for cv. Maidy. |
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Keywords: | histology in vitro PVS2 recovery rose shoot tip |
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