Abstract
Shoot tips from in vitro plants of four rose species were cryopreserved by the droplet vitrification method. Optimized conditions involved exposure to loading solution for 20 min, then treatment with plant vitrification solution (PVS2) for 20 min (<em>Rosa agrestis</em>, <em>R. canina</em> and <em>R. dumalis</em>) or 30 min (<em>R. rubiginosa</em>) followed by freezing in liquid nitrogen. Survival rate ranged from 78.3 to 95.1%, depending on the species. Regrowth rate of shoot tips was 50.5% for <em>R. agrestis</em>, 63.2% for <em>R. rubiginosa</em>, 71.4% for <em>R. dumalis</em> and 78% for <em>R. canina</em>. The preculture of donor plants in a medium with 0.25 µM sucrose facilitated the isolation of shoot tips and increased regrowth rate after cryopreservation. Plant regeneration was carried out in Murashige and Skoog medium with 1 µM 6-benzylaminopurine, 1.5 µM gibberellic acid and 0.087 M sucrose. Plants regenerated from cryopreserved shoot tips did not display morphological alterations in comparison with non-cryopreserved shoot tip – derived plants.
Highlights
Rosa is one of the major economically important genera in floriculture
In vitro precultures of donor plants or explants were applied to induce the tolerance to low temperature by using of media containing either a higher than standard concentration of carbohydrates and abscisic acid (ABA), proline or activated charcoal [3,4]
We established the efficient cryopresevation protocol for four species of Caninae rose shoot tips, and assessed the morphology of regenerated plants recovered from liquid nitrogen (LN) storage
Summary
Rosa is one of the major economically important genera in floriculture. Pentaploid roses from Caninae section (2n = 5x = 35) are a source of plant material for breeding. The specific type of meiosis in Caninae contributes to the great morphological diversity of dog roses [1]. Cryopreservation is a technique used to ensure safe and cost-efficient, long-term conservation of germplasm. Most explants require a special protection for liquid nitrogen (LN) conservation, which must be developed for each method of cryopreservation [2]. In vitro precultures of donor plants or explants were applied to induce the tolerance to low temperature by using of media containing either a higher than standard concentration of carbohydrates (sucrose, glucose) and ABA, proline or activated charcoal [3,4]
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