Efficient cryopreservation by droplet vitrification of pentaploid roses and the phenotype of regenerated plants

Bożena Pawłowska, Bożena Szewczyk-Taranek

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 (Rosa agrestis, R. canina and R. dumalis) or 30 min (R. rubiginosa) 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 R. agrestis, 63.2% for R. rubiginosa, 71.4% for R. dumalis and 78% for R. canina. 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.

Keywords


Caninae; dog roses; shoot tips; preculture; in vitro; liquid nitrogen

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References


Ritz CM, Schmuths H, Wisseman V. Evolution by reticulation: European dogroses originated by multiple hybridization across the genus Rosa. J Hered. 2005;96:4–14. http://dx.doi.org/10.1093/jhered/esi011

Panis B, Lambardi M. Status of cryopreservation technologies in plants (crops and forest trees). In: Ruane J, Sonnino A, editors. The role of biotechnology in exploring and protecting agricultural genetic resources. Rome: Food and Agriculture Organization of the United nations; 2005. p. 43–51.

Burritt DJ. Efficient cryopreservation of adventitious shoots of Begonia × erythrophylla using encapsulation–dehydration requires pretreatment with both ABA and proline. Plant Cell Tiss Organ Cult. 2008;95:209–215. http://dx.doi.org/10.1007/s11240-008-9434-5

Pawłowska B, Bach A. Cryopreservation by encapsulation dehydration of in vitro grown shoot buds of rosa ‘New Dawn’. Acta Hortic. 2011;908:303–307. http://dx.doi.org/10.17660/ActaHortic.2011.908.40

Lynch PT, Harris WC, Chartier Hollis JM. The cryopreservation of shoot tips of Rosa multiflora. Plant Growth Regul. 1996;20:43–45. http://dx.doi.org/10.1007/BF00024056

Halmagyi A, Pinker I. Plant regeneration from Rosa shoot tips cryopreserved by a combined droplet-vitrification method. Plant Cell Tiss Organ Cult. 2006;84:145–153. http://dx.doi.org/10.1007/s11240-005-9012-z

Halmagyi A, Pinker I. Cryopreservation of Rosa shoot tips: importance of preculture conditions. Acta Hortic. 2006;725:351–356. http://dx.doi.org/10.17660/ActaHortic.2006.725.45

Pawłowska B. Cryopreservation of Rosa canina and R. rubiginosa apical buds by the droplet vitrification method. Acta Hortic. 2012;937:905–910. http://dx.doi.org/10.17660/ActaHortic.2012.937.111

Le Bras CL, Besesrais PH, Hamama L, Grapin A. Cryopreservation of ex-vitro-grown Rosa chinensis ‘Old Blush’ buds using droplet-vitrification and encapsulation-dehydration. Plant Cell Tiss Organ Cult. 2014;116:235–242. http://dx.doi.org/10.1007/s11240-013-0400-5

Pawłowska B, Szewczyk-Taranek B. Droplet vitrification cryopreservation of Rosa canina and Rosa rubiginosa using shoot tips from in situ plants. Sci Hortic (Amsterdam). 2014;168:151–156. http://dx.doi.org/10.1016/j.scienta.2013.12.016

Agrawal A, Sanayaima R, Singh R, Tandon R, Verma S, Tyagi RK. Phenotypic and molecular studies for genetic stability assessment of cryopreserved banana meristems derived form field and in vitro explant sources. In Vitro Cell Dev Biol Plant. 2014;50:345–356. http://dx.doi.org/10.1007/s11627-014-9606-4

Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant. 1962;15:473–497. http://dx.doi.org/10.1111/j.1399-3054.1962.tb08052.x

Harding K. Genetic integrity of cryopreserved plant cells: a review. Cryo Letters. 2004;25:3–22.




DOI: https://doi.org/10.5586/asbp.2015.038

Journal ISSN:
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  • 0001-6977 (print; ceased since 2016)
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Polish Botanical Society