Cryopreservation of bred cultivars of rowan from the VIR in vitro collection
https://doi.org/10.30901/2658-6266-2025-3-o6
Abstract
Background. Cryopreservation is a promising strategy for the long-term preservation of genetic diversity of rowan species and cultivars in controlled environments. It minimizes the risk of genetic changes in accessions stored in liquid nitrogen. However, only a few studies on the cryopreservation of single rowan accessions are known. The objective of our study was the cryopreservation of six rowan cultivars bred in Russia and their transfer to the VIR cryobank for long-term storage. Materials and methods. Accessions of six Russian rowan cultivars (‘Alaya krupnaya’, 'Burka', 'Granatnaya', ‘Krasnaya krupnaya’, 'Sorbinka' and 'Titan') from the VIR in vitro collection, including two cultivars bred by I.V. Michurin, were taken for cryopreservation. The droplet vitrification method modified at VIR was used for cryopreservation of microplant apices. Results and discussion. The level of post-cryogenic regeneration in the control variants varied from 53% ('Granatnaya') to 97% (‘Alaya krupnaya’). The duration of in vitro cultivation of the accessions which varied from two to 17 years did not significantly affect the level of their post-cryogenic regeneration. All six cultivars were transferred to the VIR cryobank for long-term storage in the amount of 90 apices per accession. Conclusion. The droplet vitrification method modified at VIR has demonstrated high efficiency for cryopreservation of rowan accessions. Six rowan cultivars were placed in the VIR cryobank with the post-cryogenic regeneration level of 53%-97%. Further increase of the cryocollection of rowan accessions is planned.
Supporting agencies: The work was carried out as part of the implementation of the Program of Development of the National Center for Plant Genetic Resources under the Agreement No. 075-02-2025-1584 with the Ministry of Education and Science of Russia dated February 26, 2025.
About the Authors
S. E. Dunaeva
N.I. Vavilov All-Russian Institute of Plant Genetic Resources
Russian Federation
Russian Federation
Svetlana E. Dunaeva, Cand. Sci. (Biology), Senior Researcher, Department of Biotechnology, VIR
42, 44, Bolshaya Morskaya Street, St. Petersburg, 190000 Russia
O. V. Lisitsyna
N.I. Vavilov All-Russian Institute of Plant Genetic Resources
Russian Federation
Russian Federation
Olga V. Lisitsyna, Leading Specialist, Department of Biotechnology, VIR
42, 44, Bolshaya Morskaya Street, St. Petersburg, 190000 Russia
N. N. Volkova
N.I. Vavilov All-Russian Institute of Plant Genetic Resources
Russian Federation
Russian Federation
Natalia N. Volkova, Leading Specialist, Department of Biotechnology, VIR
42, 44 Bolshaya Morskaya Street, St. Petersburg, 190000 Russia
T. A. Gavrilenko
N.I. Vavilov All-Russian Institute of Plant Genetic Resources
Russian Federation
Russian Federation
Tatjana A. Gavrilenko, Dr. Sci. (Biology), Chief Researcher, Department of Biotechnology, VIR
42, 44, Bolshaya Morskaya Street, St. Petersburg, 190000 Russia
References
1. Abdullina R.G. Collection of Sorbus L. genus in the city of Ufa Botanical Garden. Hortus Botanicus. 2017;2:713-721. [in Russian]. DOI: 10.15393/j4.art.2017.4383
2. Aldasoro J.J, Aedo C., Navarro C., Garmendia F.M. The genus Sorbus (Maloideae, Rosaceae) in Europe and in North Africa: morphological analysis and systematics. Systematic Botany.1998;23:189-212.
3. Arrillaga I., Marzo T., Segura J. Micropropagation of juvenile and adult Sorbus domestica L. Plant Cell, Tissue and Organ Culture. 1991;27(3):341-348. DOI: 10.1007/BF00157600
4. Arvinte O.M., Senila L., Becze A., Amariei S. Rowanberry – a source of bioactive compounds and their biopharmaceutical properties. Plants. 2023;12(18):3225. DOI: 10.3390/plants12183225
5. Bobinaitė R., Grootaert C., Van Camp J., Šarkinas A., Liaudanskas M., Žvikas V., Viškelis P., Venskutonis P.R. Chemical composition, antioxidant, antimicrobial and antiproliferative activities of the extracts isolated from the pomace of rowanberry (Sorbus aucuparia L.). Food Research International. 2020;136:109310. DOI: 10.1016/j.foodres.2020.109310
6. Chalupa V. In vitro propagation of mature trees of Sorbus aucuparia L. and field performance of micropropagated trees. Journal of Forest Science. 2002;48:529-535. DOI: 10.17221/11923-JFS
7. Dobránszki J., Magyar-Tábori K., Jevcsák M., ÖRDöGH M., Jámbor-Benczúr E. Improving the in vitro rooting of micro-shoots of Sorbus rotundifolia ‘Bükk szépe’ by the sequential application of Humus® FW and Wuxal® Super organic and chemical fertilisers. The Journal of Horticultural Science and Biotechnology. 2012;87(5):509-513. DOI: 10.1080/14620316.2012.11512903
8. Drouet S., Doussot J., Garros L., Mathiron D., Bassard S., Favre-Réguillon A., Molinié R., Lainé É., Hano C. Selective synthesis of 3-O-palmitoyl-silybin, a new-to-nature flavonolignan with increased protective action against oxidative damages in lipophilic media. Molecules. 2018;23(10):2594. DOI: 10.3390/molecules23102594
9. Dunaeva S.E., Pendinen G.I., Antonova O.Y., Shvachko N.A., Ukhatova Y.V., Shuvalova L.E. Volkova N.N., Gavrilenko T.A. Preservation of vegetatively propagated crops in in vitro and cryo collections: methodological guidelines. (Sokhraneniye vegetativno razmnozhayemykh kul’tur v in vitro i krio kollektsiyakh: metodicheskiye ukazaniya). T.A. Gavrilenko (ed.). 2nd ed. St. Petersburg: VIR; 2017. [in Russian]
10. Dunaeva S.E., Orlova S.Yu., Tikhonova O.A., Gavrilenko T.A. In vitro collection of berry and fruit crops and their wild relatives at VIR. Plant Biotechnology and Breeding. 2018;1(1):43-51. [in Russian]. DOI: 10.30901/2658-6266-2018-1-43-51
11. Dussert S., Engelmann F., Noirot M. Development of probabilistic tools to assist in the establishment and management of cryopreserved plant germplasm collections. CryoLetters. 2003;24(3):149-160.
12. Efremova O.S., Volkova N.N., Rybakov D.А., Lisitsyna O.V., Ozerski P.V., Gavrilenko T.A. Development of the potato cryocollection preserved in the VIR cryobank. Proceedings on Applied Botany, Genetics and Breeding. 2023;184(3):9-20. [in Russian]. DOI: 10.30901/2227-8834-2023-3-9-20
13. Ermakov M.A., Volkova O.D., Khotsialova L.I., Zagumennikova T.N., Potapova A.V. Rowan in the collection of the Laboratory of Cultivated Plants of the N.V. Tsitsin Main Botanical Garden of the Russian Academy of Sciences (Ryabina v kollektsii laboratorii kulturnykh rasteniy Glavnogo botanicheskogo sada imeni N.V. Tsitsina RAN) Hortus Botanicus. 2019;14. [in Russian]. DOI: 10.15393/j4.art.2019.6224
14. Firsov G.A.,Vasiljev N.P. The genus Rowan (Sorbus) in the collection of Peter the Great Botanical Gardens in St. Petersburg. (Rod Ryabina (Sorbus) v kollektsii Botanicheskogo sada Petra Velikogo v Sankt-Peterburge). Flora and vegetation of Asian Russia. 2015;4(20):86-93. [in Russian]
15. Gavrilenko T.A., Shvachko N.A., Volkova N.N., Ukhatova Yu.V. A modified droplet vitrification method for cryopreservation of shoot tips from in vitro potato plants. Vavilov Journal of Genetics and Breeding. 2019;23(4):422-429. [in Russian]. DOI: 10.18699/VJ19.505
16. Gianguzzi V., Sottile F. Temporary immersion system as an innovative approach for in vitro propagation of Sorbus domestica L. Horticulturae 2024;10(2):164. DOI: 10.3390/horticulturae10020164
17. Goncharova O.A. Assessment of vitality and decorative qualities of introduced plants of the genus Sorbus L. on the Kola Peninsula (Otsenka zhiznennogo sostoyaniya i dekorativnykh kachestv introdutsirovannykh rasteniy roda Sorbus L. na Kol’skom poluostrove). Hortus Botanicus. 2022;17:139-153. [in Russian]
18. Gorbunov A.B. Introduction of not widespread fruits and berries in Siberia for their application as functional food products. Vestnik NSAU (Novosibirsk State Agrarian University). 2018;(4):62-73. [in Russian]. DOI: 10.31677/2072-6724-2018-49-4-62-73
19. Kadolsky M. Kryokonservierung und in vitro kultur von Pyrus pyraster (L.) Burgsd. und Sorbus torminalis (L.) Crantz . 2005. [in German] DOI: 10.18452/15652
20. Kamnev А.М., Dunaeva S.Е., Volkova N.N., Lisitsyna O.V., Gavrilenko Т.А. Cryopreservation of raspberry cultivar accessions bred in Russia from the VIR in vitro collection. Plant Biotechnology and Breeding. 2022;5(1):17-27. [in Russian]. DOI: 10.30901/2658-6266-2022-1-o2
21. King R., Bassil N.V., Rounsaville T.J., Reinhold L.A. Sorbus sensu lato: a complex genus with unfulfilled crop potential. Journal of the American Pomological Society. 2023;77(2):110-127.
22. Klein A., Bockhorn O., Mayer K., Grabner M. Central European wood species: characterization using old knowledge. Wood Science. 2016;62:194-202. DOI: 10.1007/s10086-015-1534-3
23. Komarov V.L. Rowan – Sorbus L. In the book: Flora of the USSR. Vol. 9. Moscow-Leningrad: USSR Academy of Sciences Publ.; 1939. p.372-395. [in Russian]
24. Konovalov I.N. Rowan Sorbus L. (Ryabina Sorbus L.) In: Konovalov I.N. Trees and shrubs of the USSR. Vol. 3. Moscow-Leningrad: USSR Academy of Sciences Publ.; 1954. p.458-483. [in Russian]
25. Kuminov E.P., Zhidekhina T.V. Introduction to the culture of wild fruit plants. (Vvedenie v kul’turu dikorastushchikh plodovykh rasteniy). Netraditsionnye selskokhozyaistvennye, lekarstvennye i dekorativnye rasteniya = Non-traditional agricultural, medicinal and ornamental plants. 2003;1(1):44-60. [in Russian]
26. Ludwig S., Robertson A., Rich T.C.G., Djordjevic M., Cerovic R., Houston L., Harris S.A., Hiscock S.J. Breeding systems, hybridization and continuing evolution in Avon Gorge Sorbus. Annals of Botany. 2013;111:563-575. DOI: 10.1093/aob/mct013
27. Máchová P., Malá J., Cvrčková H., Dostál J., Buriánek V. In vitro reproduction of rare and endemic species of rowan tree. The Journal of Forest Science. 2013;59:386-390. DOI: 10.17221/46/2013-JFS
28. Mándy A., Jámbor-Benczúr E., Szafián Z., Csillag A. The effect of basic media and growth regulators on in vitro propagated Sorbus degenii ‘Csákvár’. Acta Horticulturae. 1997;447:157-159. DOI: 10.17660/ActaHortic.1997.447.26
29. Mendler-Drienyovszki N., Magyar-Tábori K. Response of rowan berry (Sorbus redliana) shoot culture to slow growth storage conditions. Plants. 2023;12:1287. DOI: 10.3390/plants12061287
30. Michurin I.V. Results of sixty years of work: [1855-1935] (Itogi shestidesyatiletnikh rabot: [1855-1935]). Moscow: Selkhozgiz; 1949. [in Russian]
31. Mikulic-Petkovsek M., Krska B., Kiprovski B., Veberic R. Bioactive Components and Antioxidant Capacity of Fruits from Nine Sorbus Genotypes. The Journal of Food Science. 2017;82:647-658. DOI: 10.1111/1750-3841.13643
32. Murashige T., Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. 1962;15:473-497. DOI: 10.1111/j.1399-3054.1962.tb08052.x
33. NCGR-Corvallis Sorbus Catalog Mountain Ash. Sorbus Cultivars, Selections, and Hybrids. Available from: https://www.ars.usda.gov/ARSUserFiles/20721500/catalogs/soracc.html [accessed Sept. 15, 2025].
34. Panis B. Sixty years of plant cryopreservation: from freezing hardy mulberry twigs to establishing reference crop collections for future generations. Acta Horticulture. 2019;1234:1-7. DOI: 10.17660/ActaHortic.2019.1234.1
35. Panis B., Piette B., André E., Van den Houwe I., Swennen R. Droplet vitrification: the first generic cryopreservation protocol for organized plant tissues? Acta Horticulture. 2011;908:157-162. DOI: 10.17660/ActaHortic.2011.908.17
36. Panis B., Piette B., Swennen R. Droplet vitrification of apical meristems: A cryopreservation protocol applicable to all Musaceae. Plant Science. 2005;168:45-55. DOI: 10.1016/j.plantsci.2004.07.022
37. Petrov E.M. Rowan (Ryabina). Moscow: Selkhogiz; 1957. [in Russian]
38. Phipps J.B., Robertson K., Smith P.G., Rohrer J.R. A checklist of the subfamily Maloideae (Rosaceae). Canadian Journal of Botany. 1990;68:2209-2269. DOI: 10.1139/b90-288
39. Poplavskaya T.K. Breeding and introduction of new rowan varieties in Russian horticulture (Selektsiya i vnedreniye novykh sortov ryabiny v sadovodstvo Rossii). Perm: Perm Book Publishing House; 2006. [in Russian]
40. Raudonis R., Raudonė L., Gaivelytė K., Viškelis P., Janulis V. Phenolic and antioxidant profiles of rowan (Sorbus L.) fruits. Natural Products Research. 2014;28(16):1231-1240. DOI: 10.1080/14786419.2014.895727.
41. Rengarten G.A., Sorokopudov V.N. Selection of rows as a decorative culture in Russia and in European countries. the Bulletin of the KrasGAU. 2019a;6(147):9-15. [in Russian]. URL: http://www.kgau.ru/vestnik/2019_6/content/2.pdf [дата обращения: 24.09.2025]
42. Rengarten G.A., Sorokopudov V.N. Introduction and breeding of Sorbus (Rosaceae) as a food plant in countries of the world (Introduktsiya i selektsiya Sorbus (Rosaceae) v kachestve pishchevogo rasteniya v stranakh mira). Ekosistemy = Ecosystems. 2019b;18(48):89-96. [in Russian]
43. Sarv V, Venskutonis P.R, Bhat R. The Sorbus spp. – Underutilised plants for foods and nutraceuticals: review on polyphenolic phytochemicals and antioxidant potential. Antioxidants (Basel). 2020;Sep.1;9(9):813. DOI: 10.3390/antiox9090813
44. Šedivá J.; Velebil J.; Zahradník D. Micropropagation as a tool for the conservation of autochthonous Sorbus species of Czechia. Plants. 2023;12:488. DOI: 10.3390/plants 12030488
45. Sedov E.N. et al. (eds). Pomology. Vol. 5: Strawberry, raspberry, nut and rare crops (Pomologiya. T. 5: Zemlyanika, malina, orekhoplodnyie i redkiye kul’tury). Orel: VNIISPB; 2014. [in Russian]
46. Sokolov V.A., Savel’ev N.I., Goncharov N.P. I.V. Michurin’s work on expansion of the plant horticulture assortment and improvement of food quality. Proceedings of the Latvian Academy of Sciences. 2015;69(4):90-197. DOI: 10.1515/prolas-2015-0028
47. State Register of Varieties and Hybrids of Agricultural Plants Admitted for Usage (National List): official publication. Moscow: Rosinformagrotech; 2024. [in Russian]. URL: https://gossortrf.ru/upload/iblock/00a/clri6obhudueqx6t1f6awcrsp6vm6psk.pdf [дата обращения: 15.09.2025]
48. Streltsina S.A Burmistrov L.A., Nikitina E.V. Nutrient and bioactive substances in rowan (Sorbus L.) berries in the conditions of the northwestern horticultural zone of Russia (Pitatelnyie i biologicheski aktivnyie veshchestva plodov ryabiny (Sorbus L.) v usloviyakh Severo-Zapadnoy zony sadovodstva Rossii). Agrarnaya Rossiya = Agrarian Russia. 2010;3:10-17. [in Russian]
49. Taran A.A., Taran An.A., Chabanenko S.I., Sheiko V.V., Kazhaeva M.V. Catalogue of plants of the Sakhalin Botanical Garden FEB RAS: reference manual (Katalog rasteniy Sakhalinskogo botanicheskogo sada DVO RAN: spravochnoye posobie). V.M. Eremin (ed.). Yuzhno-Sakhalinsk: SB BSI FEB RAS; 2011. [in Russian]
50. The European Red List of Trees. Available from: https://www.bgci.org/our-work/projects-and-case-studies/the-red-list-of-european-trees/ [accessed Sept. 15, 2025].
51. Towill L.E. Improved survival after cryogenic exposure of shoot tips derived from in vitro plantlet cultures of potato. Cryobiology. 1983;20(5):567-573. DOI: 10.1016/0011-2240(83)90045-7
52. Ukhatova Y.V., Dunaeva S.E., Antonova O.Y., Apalikova O.V., Pozdniakova K.S., Novikova L.Y., Shuvalova L.E., Gavrilenko T.A. Cryopreservation of red raspberry cultivars from the VIR in vitro collection using a modified droplet vitrification method. In Vitro Cellular and Developmental Biology – Plant. 2017;53(4):394-401. DOI: 10.1007/s11627-017-9860-3
53. Vitkovsky V.L. Fruit Plants of the World. St. Petersburg, Moscow, Krasnodar: Lan’; 2003. [in Russian] (Витковский В.Л. Плодовые растения мира. Санкт-Петербург, Москва, Краснодар: Лань; 2003).
54. Volk G.M., Henk A.D., Jenderek M.M., Richards C.M. Probabilistic viability calculations for cryopreserving vegetatively propagated collections in genebanks. Genetic Resources and Crop Evolution. 2017;64(7):1613-1622. DOI: 10.1007/s10722-016-0460-6
55. Vysotskaya O.N. Cryopreservation in vitro of mountain ash (Sorbus L.) shoot tips. In: The IX International Conference «The Biology of plant cells in vitro and biotechnology»: Abstract; 2008 September 8-12; Zvenigorod, Russia. Moscow; 2008. p. 80-81. [in Russian]. URL: https://ippras.ru/upload/files/Abstracts.pdf [дата обращения 15.09.2025].
56. Yuorieva N.; Sinetova M.; Messineva E., Kulichenko I., Fomenkov A., Vysotskaya O., Osipova E., Baikalova A., Prudnikova O., Titova M., Nosov A.V., Popova E. Plants, cells, algae, and cyanobacteria in vitro and cryobank collections at the Institute of Plant Physiology, Russian Academy of Sciences – a platform for research and production center. Biology. 2023;12(6):838. DOI: 10.3390/biology12060838
Review
For citations:
Dunaeva S.E., Lisitsyna O.V., Volkova N.N., Gavrilenko T.A. Cryopreservation of bred cultivars of rowan from the VIR in vitro collection. Plant Biotechnology and Breeding. 2025;8(3):55-64. (In Russ.) https://doi.org/10.30901/2658-6266-2025-3-o6
Views:
49
Email this article 