- PII
- S3034546425040029-1
- DOI
- 10.7868/S3034546425040029
- Publication type
- Article
- Status
- Published
- Authors
- Volume/ Edition
- Volume 94 / Issue number 4
- Pages
- 330-340
- Abstract
- The efficiency of cryopreservation of cyanobacteria and eukaryotic microalgae of different taxonomic groups, including charophyte, chlorophyte and ochrophyte microalgae, was assessed using 24 strains of the algological part of the All-Russian Collection of Microorganisms (VKM) as an example. Microalgae cultures differing in cell size, morphological organization of the thallus, reproduction method, presence and size of mucous membranes, and ability to form dormant cells were selected for comparative study. Two types of cryoprotectors (methanol and dimethyl sulfoxide), two types of nutrient media (slant agar and liquid medium), and three methods for determining the survival of microalgae after cryopreservation were tested. It was shown that a two-step cryopreservation protocol using both cryoprotectants (methanol and dimethyl sulfoxide) was successfully applied to all 24 studied strains, regardless of their taxonomic affiliation and morphological features. Based on the results of the experiment, a standard operating procedure for cryopreservation of microalgae was developed, including a liquid culture medium with dimethyl sulfoxide, as well as two methods for determining the survival of microalgae after cryopreservation - growth in a liquid medium and streaks on agar. The proposed protocol not only ensures the preservation of cell viability and the possibility of further use of the strain as a morphologically and genetically representative sample, but also minimizes time and resource costs, as well as the risk of contamination of cultures.
- Keywords
- цианобактерии микроводоросли длительное хранение выживаемость стратегия ex situ биоресурсная коллекция
- Date of publication
- 01.08.2025
- Year of publication
- 2025
- Number of purchasers
- 0
- Views
- 66
References
- 1. Beaty M.H. Cryopreservation of eukaryote algae. Master of Science in Biology thesis. Blacksburg, Virginia: Virginia Polytechnic Institute and State University, 1991. 116 p.
- 2. Cameron R. E. Species of Nostoc vaucher occurring in the Sonoran Desert in Arizona // Trans. Am. Microsc. Soc. 1962. V. 81. P. 379-384. https://doi.org/10.2307/3223790
- 3. Day J. G., Priestley I. M., Codd G. A. Storage, recovery and photosynthetic activities of immobilised algae // Plant and animal cells: process possibilities / Eds. Webb C., Mavituna F. Chichester, West Sussex: Ellis Horwood Limited, 1987. P. 257-261.
- 4. Ettl H., Gärtner G. Syllabus der boden-, luft-und flech-tenalgen. 2., ergänzte Auflage. Berlin, Heidelberg: Springer Spektrum, 2014. 773 p. https://doi.org/10.1007/978-3-642-39462-1
- 5. Fernandes M. S., Calsing L. C., Nascimento R. C., Santana H., Morais P. B., de Capdeville G., Brasil B. S. Customized cryopreservation protocols for chlorophytes based on cell morphology // Algal Res. 2019. V. 38. Art. 101402. https://doi.org/10.1016/j.algal.2018.101402
- 6. Foo S. C., Mok C. Y., Ho S. Y., Khong N. M.H. Microalgal culture preservation: progress, trends and future developments // Algal Res. 2023. V. 71. Art. 103007. https://doi.org/10.1016/j.algal.2023.103007
- 7. Friedl T., Lorenz M. The culture collection of algae at Göttingen University (SAG): a biological resource for biotechnological and biodiversity research // Procedia Environ. Sci. 2012. V. 15. P. 110-117. https://doi.org/10.1016/J.PROENV.2012.05.015
- 8. Guiry M. D. How many species of algae are there? A reprise. Four kingdoms, 14 phyla, 63 classes and still growing // J. Phycol. 2024. V. 60. P. 214-228. https://doi.org/10.1111/jpy.13431
- 9. Holm-Hansen O. Viability of blue-green and green algae after freezing // Physiol. Plant. 1963. V. 16. P. 530-540. https://doi.org/10.1111/j.1399-3054.1963.tb08330.x
- 10. Kapoore R. V., Huete-Ortega M., Day J. G., Okurowska K., Slocombe S. P., Stanley M. S., Vaidyanathan S. Effects of cryopreservation on viability and functional stability of an industrially relevant alga // Sci. Rep. 2019. V. 9. Art. 2093. https://doi.org/10.1038/s41598-019-38588-6
- 11. Leeson E. A., Cann J. P., Morris G. J. Maintenance of algae and protozoa // Maintenance of Microorganisms / Eds. Kirsop B.E., Snell J. J.S. London: Academic Press, 1984. P. 131-160.
- 12. Lewis L. A., Trainor F. R. Survival of Protosiphon botryoides (Chlorophyceae, Chlorophyta) from a Connecticut soil dried for 43 years // Phycologia. 2012. V. 51. P. 662-665. https://doi.org/10.2216/11-108.1
- 13. Leya T. The CCCryo Culture Collection of Cryophilic Algae as a valuable bioresource for algal biodiversity and for novel, industrially marketable metabolites // Appl. Phycol. 2022. V. 3. P. 167-188. https://doi.org/10.1080/26388081.2020.1753572
- 14. Prieto-Guevara M., Alarcón-Furnieles J., Jiménez-Velásquez C., Hernández-Julio Y., Espinosa-Araujo J., Atencio-García V. Cryopreservation of the microalgae Scenedesmus sp. // Cells. 2023. V. 12. Art. 562. https://doi.org/10.3390/cells12040562
- 15. Morris G. J. Cryopreservation of 250 strains of Chlorococ-cales by the method of two-step cooling // Br. Phycol. J. 1978. V. 13. P. 15-24. https://doi.org/10.1080/00071617800650031
- 16. Müller J., Day J. G., Harding K., Hepperle D., Lorenz M., Friedl T. Assessing genetic stability of a range of terrestrial microalgae after cryopreservation using amplified fragment length polymorphism (AFLP) // Am.J. Bot. 2007. V. 94. P. 799-808. https://doi.org/10.3732/ajb.94.5.799
- 17. Osório H. C., Laranjeiro C. N., Santos L. M., Santos M. F. First attempts to cryopreserve strains from the Coimbra Collection of Algae (ACOI) and the use of image analysis to assess viability // Nova Hedwigia. 2004. V. 79. P. 227-235. https://doi.org/10.1127/0029-5035/2004/0079-0227
- 18. Paredes E., Ward A., Probert I., Gouhier L., Campbell C. N. Cryopreservation of Algae // Cryopreservation and freeze-drying protocols. Methods in Molecular Biology / Eds. Wolkers W. F., Oldenhof H. NY: Humana New York, 2021. V. 2180. P. 607-621. https://doi.org/10.1007/978-1-0716-0783-1_32
- 19. Puchkov E. O. Preservation of viable microorganisms in the laboratory: an overview of basics, methods and practical recommendations for beginners // Austin J. Biotechnol. Bioeng. 2023. V. 10. Art. 1119. https://doi.org/10.26420/austinjbiotechnolbioeng.2023.1119
- 20. Rastoll M. J., Ouahid Y., Martín-Gordillo F., Ramos V., Vasconcelos V., Del Campo F. F. The development of a cryopreservation method suitable for a large cyanobacteria collection // J. Appl. Phycol. 2013. V. 25. № 5. P. 1483-1493. https://doi.org/10.1007/s10811-013-0001-z
- 21. Urmeneta J., Navarrete A., Huete J., Guerrero R. Isolation and characterization of cyanobacteria from microbial mats of the Ebro Delta, Spain // Curr. Microbiol. 2003. V. 46. P. 0199-0204. https://doi.org/10.1007/s00284-002-3856-9
- 22. Wan M. C., Qin W., Lei C., Li Q. H., Meng M., Fang M., Song W., Chen J. H., Tay F., Niu L. N. Biomaterials from the sea: Future building blocks for biomedical applications // Bioact. Mater. 2021. V. 6. P. 4255-4285. https://doi.org/10.1016/j.bioactmat.2021.04.028
