{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T16:04:58Z","timestamp":1781625898403,"version":"3.54.5"},"reference-count":62,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T00:00:00Z","timestamp":1764547200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T00:00:00Z","timestamp":1768176000000},"content-version":"vor","delay-in-days":42,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Simmondsia chinensis<\/jats:italic>\n                    , commonly known as jojoba, is an important renewable source of liquid wax esters valued for its unique seed oil that is used in several industries. Elite jojoba germplasm needs to be conserved owing to its overexploitation, climate variability and excessive dependence of selected cultivars. This study presents a straightforward and efficient droplet vitrification-based cryopreservation protocol in jojoba. Shoot tips isolated from eight-week-old cultures were cultured for two days on high sucrose (0.3\u00a0M) enriched medium and then treated with loading solution containing 0.4\u00a0M sucrose and 2\u00a0M glycerol, followed by PVS2 exposure for 30\u00a0min at room temperature. Vitrified shoot tips were then directly frozen in liquid nitrogen by placing them on aluminium foil strips. Frozen shoot tips were rewarmed in an unloading solution containing 1.2\u00a0M sucrose for 15\u00a0min, and then cultured on regeneration medium consisting of Murashige and Skoog medium (MS) supplemented with 4.97\u00b5M benzyl-amino purine (BAP) and 0.28 \u00b5M gibberellic acid (GA\n                    <jats:sub>3<\/jats:sub>\n                    ). The protocol was optimized in one accession, where as high as 85.7% post-thaw survival and 76.1% regrowth were observed. The developed protocol was then tested for its efficacy and reproducibility on eleven other accession, and high post-thaw regrowth, ranging from 50 to 76.13% was observed. This study presents a broad spectrum, reproducible and efficient protocol for the conservation of jojoba genetic resources. This is the first report on cryopreservation of jojoba germplasm for its long-term conservation, providing a technical platform to set up cryobanks of valuable material of this important commercial crop.\n                  <\/jats:p>","DOI":"10.1038\/s41598-025-30951-0","type":"journal-article","created":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T19:05:13Z","timestamp":1764615913000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An efficient and reproducible cryopreservation protocol for sustainable conservation of Jojoba (Simmondsia chinensis)"],"prefix":"10.1038","volume":"16","author":[{"given":"Era Vaidya","family":"Malhotra","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suresh Chand","family":"Mali","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sangita","family":"Bansal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anju Mahendru","family":"Singh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,12,1]]},"reference":[{"key":"30951_CR1","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1111\/j.1365-313X.2008.03430.x","volume":"54","author":"JM Dyer","year":"2008","unstructured":"Dyer, J. M., Stymne, S., Green, A. G. & Carlsson, A. S. High value oils from plants. Plant. J. 54, 640\u2013655 (2008).","journal-title":"Plant. J."},{"key":"30951_CR2","doi-asserted-by":"publisher","first-page":"1836","DOI":"10.1016\/j.renene.2019.09.139","volume":"147","author":"S Agarwal","year":"2020","unstructured":"Agarwal, S., Kumari, S., Mudgal, A. & Khan, S. Green synthesized nano additives in Jojoba biodiesel diesel blends: an improvement of engine performance and emission. Renew. Energy. 147, 1836\u20131844 (2020).","journal-title":"Renew. Energy"},{"key":"30951_CR3","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.aoas.2018.04.003","volume":"63","author":"S Agarwal","year":"2018","unstructured":"Agarwal, S., Arya, D. & Khan, S. Comparative fatty acid and trace elemental analysis identified the best Raw material of Jojoba (Simmondsia chinensis) for commercial applications. Ann. Agric. Sci. 63, 37\u201345 (2018).","journal-title":"Ann. Agric. Sci."},{"key":"30951_CR4","doi-asserted-by":"crossref","unstructured":"Hill, K. & Hofer, R. Natural fats and oils. In Sustainable Solutions for Modern Economies (ed Hofer, R.) 167\u2013237 (Royal Society of Chemistry, 2009).","DOI":"10.1039\/9781847552686-00167"},{"key":"30951_CR5","doi-asserted-by":"publisher","first-page":"987","DOI":"10.1002\/ptr.1363","volume":"17","author":"T Aburjai","year":"2003","unstructured":"Aburjai, T. & Natsheh, F. M. Plants used in cosmetics. Phytother Res. 17, 987\u20131000 (2003).","journal-title":"Phytother Res."},{"key":"30951_CR6","doi-asserted-by":"publisher","first-page":"268","DOI":"10.1111\/j.1473-2165.2008.00405.x","volume":"7","author":"J Meyer","year":"2008","unstructured":"Meyer, J., Marshall, B., Gacula, M. Jr. & Rheins, L. Evaluation of additive effects of hydrolyzed Jojoba (Simmondsia chinensis) esters and glycerol: A preliminary study. J. Cosmet. Dermatol. 7, 268\u2013274 (2008).","journal-title":"J. Cosmet. Dermatol."},{"key":"30951_CR7","doi-asserted-by":"publisher","first-page":"443","DOI":"10.1016\/j.jep.2010.12.042","volume":"134","author":"E Ranzato","year":"2011","unstructured":"Ranzato, E., Martinotti, S. & Burlando, B. Wound healing properties of Jojoba liquid wax: an in vitro study. J. Ethnopharmacol. 134, 443\u2013449 (2011).","journal-title":"J. Ethnopharmacol."},{"key":"30951_CR8","doi-asserted-by":"publisher","first-page":"765","DOI":"10.1007\/BF02663058","volume":"56","author":"T Miwa","year":"1979","unstructured":"Miwa, T., Rothfus, J. A. & Dimitroff, E. Extreme pressure lubricant tests on Jojoba and sperm Whale oils. J. Am. Oil Chem. Soc. 56, 765\u2013770 (1979).","journal-title":"J. Am. Oil Chem. Soc."},{"key":"30951_CR9","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1016\/0043-1648(93)90469-3","volume":"161","author":"RPS Bisht","year":"1993","unstructured":"Bisht, R. P. S., Sivasankaran, G. A. & Bhatia, V. K. Additive properties of Jojoba oil for lubricating oil formulations. Wear 161, 193\u2013197 (1993).","journal-title":"Wear"},{"key":"30951_CR10","doi-asserted-by":"publisher","first-page":"639","DOI":"10.1080\/00908310490438623","volume":"26","author":"O El Kinawy","year":"2004","unstructured":"El Kinawy, O. Comparison between Jojoba oil and other vegetable oils as a substitute to petroleum. Energy Sources. 26, 639\u2013645 (2004).","journal-title":"Energy Sources"},{"key":"30951_CR11","unstructured":"Wisniak, J. The chemistry and technology of Jojoba oil. Am. Oil Chem. Soc. (1987)."},{"key":"30951_CR12","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1186\/s40659-017-0131-x","volume":"50","author":"JR Al Obaidi","year":"2017","unstructured":"Al Obaidi, J. R. et al. A review on plant importance, biotechnological aspects, and cultivation challenges of Jojoba plant. Biol. Res. 50, 50 (2017).","journal-title":"Biol. Res."},{"key":"30951_CR13","first-page":"298","volume":"4","author":"RK Tyagi","year":"2001","unstructured":"Tyagi, R. K. & Prakash, S. Clonal propagation and in vitro conservation of Jojoba (Simmondsia chinensis). Indian J. Plant. Genet. Resour. 4, 298\u2013300 (2001).","journal-title":"Indian J. Plant. Genet. Resour."},{"key":"30951_CR14","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1023\/B:BIOP.0000024270.02186.1f","volume":"48","author":"RK Tyagi","year":"2004","unstructured":"Tyagi, R. K. & Prakash, S. Genotype and sex specific protocols for in vitro micropropagation and medium term conservation of Jojoba. Biol. Plant. 48, 19\u201323 (2004).","journal-title":"Biol. Plant."},{"key":"30951_CR15","first-page":"396","volume":"5","author":"SA Bekheet","year":"2016","unstructured":"Bekheet, S. A. et al. In vitro conservation of Jojoba (Simmondsia chinensis) shootlet cultures using osmotic stress and low temperature. Middle East. J. Agric. Res. 5, 396\u2013402 (2016).","journal-title":"Middle East. J. Agric. Res."},{"key":"30951_CR16","doi-asserted-by":"publisher","first-page":"583","DOI":"10.1016\/j.biotechadv.2014.03.003","volume":"32","author":"B Wang","year":"2014","unstructured":"Wang, B. et al. Potential applications of cryogenic technologies to plant genetic improvement and pathogen eradication. Biotechnol. Adv. 32, 583\u2013395 (2014).","journal-title":"Biotechnol. Adv."},{"key":"30951_CR17","doi-asserted-by":"publisher","first-page":"1","DOI":"10.17660\/ActaHortic.2019.1234.1","volume":"1234","author":"B Panis","year":"2019","unstructured":"Panis, B. Sixty years of plant cryopreservation: from freezing hardy mulberry twigs to Establishing reference crop collections for future generations. Acta Hortic. 1234, 1\u20138 (2019).","journal-title":"Acta Hortic."},{"key":"30951_CR18","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.cryobiol.2019.01.008","volume":"8","author":"MN Normah","year":"2019","unstructured":"Normah, M. N., Sulong, N. & Reed, B. M. Cryopreservation of shoot tips of recalcitrant and tropical species: advances and strategies. Cryobiology 8, 1\u201314 (2019).","journal-title":"Cryobiology"},{"key":"30951_CR19","doi-asserted-by":"publisher","first-page":"2744","DOI":"10.3390\/plants10122744","volume":"10","author":"C Benelli","year":"2021","unstructured":"Benelli, C. Plant cryopreservation: A look at the present and the future. Plants 10, 2744 (2021).","journal-title":"Plants"},{"key":"30951_CR20","unstructured":"Acker, J. P., Adkins, S., Alves, A., Horna, D. & Toll, J. Feasibility Study for a Safety Backup Cryopreservation Facility (Bioversity International, 2017)."},{"key":"30951_CR21","first-page":"1634","volume":"9","author":"B Panis","year":"2020","unstructured":"Panis, B. & Nagel, M. Van Den Houwe, I. Challenges and prospects for the conservation of crop genetic resources in field genebanks, in in vitro collections and\/or in liquid nitrogen. Plants (Basel). 9, 1634 (2020).","journal-title":"Plants (Basel)"},{"key":"30951_CR22","doi-asserted-by":"publisher","first-page":"369","DOI":"10.1007\/s11240-012-0245-3","volume":"112","author":"CH Feng","year":"2013","unstructured":"Feng, C. H. et al. Duration of sucrose preculture is critical for shoot regrowth of in vitro grown Apple shoot tips cryopreserved by encapsulation dehydration. Plant. Cell. Tissue Organ. Cult. 112, 369\u2013378 (2013).","journal-title":"Plant. Cell. Tissue Organ. Cult."},{"key":"30951_CR23","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1016\/0304-4157(88)90015-9","volume":"947","author":"JH Crowe","year":"1989","unstructured":"Crowe, J. H. et al. Interactions of sugars with membranes. Biochim. Biophys. Acta. 947, 367\u2013384 (1989).","journal-title":"Biochim. Biophys. Acta"},{"key":"30951_CR24","doi-asserted-by":"publisher","first-page":"1073","DOI":"10.1093\/aob\/mcl054","volume":"97","author":"M Suzuki","year":"2006","unstructured":"Suzuki, M. et al. Physiological changes in Gentian axillary buds during two-step preculturing with sucrose that conferred high levels of tolerance to desiccation and cryopreservation. Ann. Bot. 97, 1073\u20131081 (2006).","journal-title":"Ann. Bot."},{"key":"30951_CR25","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1007\/s002990100347","volume":"20","author":"RA Shibli","year":"2001","unstructured":"Shibli, R. A., Haagenson, D. M., Cunningham, S. M., Berg, W. K. & Volenec, J. J. Cryopreservation of alfalfa (Medicago sativa L.) cells by encapsulation dehydration. Plant. Cell. Rep. 20, 445\u2013450 (2001).","journal-title":"Plant. Cell. Rep."},{"key":"30951_CR26","doi-asserted-by":"crossref","unstructured":"Hoekstra, F. A., Golovina, E. A., Tetteroo, F. A. A. & Wolkers, W. F. Induction of desiccation tolerance in plant somatic embryos: How exclusive is the protective role of sugars? Cryobiology 43, 140\u2013150 (2001).","DOI":"10.1006\/cryo.2001.2358"},{"key":"30951_CR27","first-page":"320","volume":"30","author":"HH Kim","year":"2009","unstructured":"Kim, H. H. et al. Development of alternative plant vitrification solutions in droplet vitrification procedures. CryoLetters 30, 320\u2013334 (2009).","journal-title":"CryoLetters"},{"key":"30951_CR28","first-page":"202","volume":"30","author":"I Pinker","year":"2009","unstructured":"Pinker, I., Halmagyi, A. & Olbricht, K. Effects of sucrose preculture on cryopreservation by droplet vitrification of strawberry cultivars and morphological stability of cryopreserved plants. CryoLetters 30, 202\u2013211 (2009).","journal-title":"CryoLetters"},{"key":"30951_CR29","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1016\/j.sajb.2010.10.005","volume":"77","author":"XL Chen","year":"2011","unstructured":"Chen, X. L. et al. X. Cryopreservation of in vitro grown apical meristems of Lilium by droplet vitrification. S Afr. J. Bot. 77, 397\u2013403 (2011).","journal-title":"S Afr. J. Bot."},{"key":"30951_CR30","doi-asserted-by":"publisher","first-page":"401","DOI":"10.5511\/plantbiotechnology.11.0531a","volume":"28","author":"K Sekizawa","year":"2011","unstructured":"Sekizawa, K., Yamamoto, S. I., Rafique, T., Fukui, K. & Niino, T. Cryopreservation of in vitro grown shoot tips of carnation (Dianthus caryophyllus L.) by vitrification method using aluminium cryo plates. Plant. Biotechnol. 28, 401\u2013405 (2011).","journal-title":"Plant. Biotechnol."},{"key":"30951_CR31","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1007\/s11627-015-9704-y","volume":"51","author":"JM Zhang","year":"2015","unstructured":"Zhang, J. M. et al. Identification of a highly successful cryopreservation method (droplet vitrification) for Petunia. Vitro Cell. Dev. Biol. -Plant. 51, 445\u2013451 (2015).","journal-title":"Vitro Cell. Dev. Biol. -Plant"},{"key":"30951_CR32","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1016\/j.scienta.2017.08.016","volume":"226","author":"XX Liu","year":"2017","unstructured":"Liu, X. X., Mou, S. W., Wen, Y. B. & Cheng, Z. H. Establishment of a Garlic cryopreservation protocol for shoot apices from adventitious buds in vitro. Sci. Hortic. 226, 10\u201318 (2017).","journal-title":"Sci. Hortic."},{"key":"30951_CR33","doi-asserted-by":"crossref","unstructured":"Zhang, A. L. et al. Overcoming challenges for shoot tip cryopreservation of root and tuber crops. Agronomy 13 (2023).","DOI":"10.3390\/agronomy13010219"},{"key":"30951_CR34","first-page":"375","volume":"25","author":"BJ Fuller","year":"2004","unstructured":"Fuller, B. J. & Cryoprotectants The essential antifreezes to protect life in the frozen state. CryoLetters 25, 375\u2013388 (2004).","journal-title":"CryoLetters"},{"key":"30951_CR35","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1016\/j.cryobiol.2005.09.004","volume":"52","author":"GM Volk","year":"2006","unstructured":"Volk, G. M. & Walters, C. Plant vitrification solution 2 lowers water content and alters freezing behavior in shoot tips during cryoprotection. Cryobiology 52, 48\u201361 (2006).","journal-title":"Cryobiology"},{"key":"30951_CR36","doi-asserted-by":"crossref","unstructured":"Sakai, A., Hirai, D. & Niino, T. Development of PVS-based vitrification and encapsulation\u2013vitrification protocols. In Plant Cryopreservation: A Practical Guide (ed Reed, B. M.) 33\u201357 (Springer, 2008).","DOI":"10.1007\/978-0-387-72276-4_3"},{"key":"30951_CR37","doi-asserted-by":"publisher","first-page":"330","DOI":"10.1016\/j.scienta.2014.07.031","volume":"176","author":"RR Wang","year":"2014","unstructured":"Wang, R. R. et al. Shoot recovery and genetic integrity of Chrysanthemum morifolium shoot tips following cryopreservation by droplet-vitrification. Sci. Hortic. 176, 330\u2013339 (2014).","journal-title":"Sci. Hortic."},{"key":"30951_CR38","first-page":"14","volume":"31","author":"AM Medina","year":"2010","unstructured":"Medina, A. M., Casas, L., Swennen, R. & Panis, B. Cryopreservation of Thymus moroderi by droplet vitrification. CryoLetters 31, 14\u201323 (2010).","journal-title":"CryoLetters"},{"key":"30951_CR39","first-page":"418","volume":"33","author":"GM Volk","year":"2012","unstructured":"Volk, G. M., Bonnart, R., Krueger, R. & Lee, R. R. Cryopreservation of citrus shoot tips using micrografting for recovery. CryoLetters 33, 418\u2013426 (2012).","journal-title":"CryoLetters"},{"key":"30951_CR40","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/plants10020277","volume":"10","author":"C Benelli","year":"2021","unstructured":"Benelli, C., Carvalho, L. S. O., Merzougui, S. E. L. & Petruccelli, R. Two advanced cryogenic procedures for improving Stevia rebaudiana (Bertoni) cryopreservation. Plants 10, 1 (2021).","journal-title":"Plants"},{"key":"30951_CR41","doi-asserted-by":"publisher","first-page":"112329","DOI":"10.1016\/j.scienta.2023.112329","volume":"321","author":"R Gowthami","year":"2023","unstructured":"Gowthami, R., Chander, S., Pandey, R., Shankar, M. & Agrawal, A. Development of efficient and sustainable droplet vitrification cryoconservation protocol for shoot tips for long term conservation of Dahlia germplasm. Sci. Hortic. 321, 112329 (2023).","journal-title":"Sci. Hortic."},{"key":"30951_CR42","doi-asserted-by":"publisher","first-page":"104887","DOI":"10.1016\/j.cryobiol.2024.104887","volume":"115","author":"EV Malhotra","year":"2024","unstructured":"Malhotra, E. V., Mali, S. C., Sharma, S. & Bansal, S. A droplet vitrification cryopreservation protocol for conservation of hops (Humulus lupulus) genetic resources. Cryobiology 115, 104887. https:\/\/doi.org\/10.1016\/j.cryobiol.2024.104887 (2024).","journal-title":"Cryobiology"},{"key":"30951_CR43","doi-asserted-by":"publisher","first-page":"117","DOI":"10.1007\/s11240-014-0585-2","volume":"120","author":"A Panta","year":"2015","unstructured":"Panta, A. et al. Improved cryopreservation method for the long term conservation of the world potato germplasm collection. Plant. Cell. Tiss Organ. Cult. 120, 117\u2013125. https:\/\/doi.org\/10.1007\/s11240-014-0585-2 (2015).","journal-title":"Plant. Cell. Tiss Organ. Cult."},{"key":"30951_CR44","doi-asserted-by":"publisher","first-page":"333","DOI":"10.5010\/JPB.2018.45.4.333","volume":"45","author":"K Nakkanong","year":"2018","unstructured":"Nakkanong, K. & Nualsri, C. Cryopreservation of Hevea Brasiliensis zygotic embryos by vitrification and encapsulation dehydration. J. Plant. Biotechnol. 45, 333\u2013339 (2018).","journal-title":"J. Plant. Biotechnol."},{"key":"30951_CR45","first-page":"71","volume":"58","author":"JC Bettoni","year":"2019","unstructured":"Bettoni, J. C., Bonnart, R., Shepherd, A., Kretzschmar, A. A. & Volk, G. M. Cryopreservation of grapevine (Vitis spp.) shoot tips from growth chamber-sourced plants and histological observations. Vitis 58, 71\u201378 (2019).","journal-title":"Vitis"},{"key":"30951_CR46","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1016\/0168-9452(93)90189-7","volume":"91","author":"S Nishizawa","year":"1993","unstructured":"Nishizawa, S., Sakai, A., Amano, Y. & Matsuzawa, T. Cryopreservation of asparagus (Asparagus officinalis L.) embryogenic suspension cells and subsequent plant regeneration by vitrification. Plant. Sci. 91, 67\u201373. https:\/\/doi.org\/10.1016\/0168-9452(93)90189-7 (1993).","journal-title":"Plant. Sci."},{"key":"30951_CR47","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1007\/s11240-019-01721-4","volume":"140","author":"MR Wang","year":"2020","unstructured":"Wang, M. R. et al. Droplet vitrification for shoot tip cryopreservation of Shallot (Allium Cepa var. aggregatum): effects of PVS3 and PVS2 on shoot regrowth. Plant. Cell. Tiss Organ. Cult. 140, 185\u2013195 (2020).","journal-title":"Plant. Cell. Tiss Organ. Cult."},{"key":"30951_CR48","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1007\/s11627-023-10399-5","volume":"60","author":"T Vujovi\u0107","year":"2024","unstructured":"Vujovi\u0107, T., An\u0111eli\u0107, T., Markovi\u0107, Z., Gajdo\u0161ov\u00e1, A. & Hunkov\u00e1, J. Cryopreservation of highbush blueberry, strawberry, and Saskatoon using V and D cryo-plate methods and monitoring of multiplication ability of regenerated shoots. Vitro Cell. Dev. Biol. -Plant. 60, 85\u201397 (2024).","journal-title":"Vitro Cell. Dev. Biol. -Plant"},{"key":"30951_CR49","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1007\/s11240-020-01846-x","volume":"144","author":"JC Bettoni","year":"2021","unstructured":"Bettoni, J. C., Bonnart, R. & Volk, G. M. Challenges in implementing plant shoot tip cryopreservation technologies. Plant. Cell. Tiss Organ. Cult. 144, 21\u201334. https:\/\/doi.org\/10.1007\/s11240-020-01846-x (2021).","journal-title":"Plant. Cell. Tiss Organ. Cult."},{"key":"30951_CR50","doi-asserted-by":"publisher","first-page":"2623","DOI":"10.3390\/plants10122623","volume":"10","author":"J Z\u00e1me\u010dn\u00edk","year":"2021","unstructured":"Z\u00e1me\u010dn\u00edk, J., Faltus, M. & Bilav\u010d\u00edk, A. Vitrification solutions for plant cryopreservation: modification and properties. Plants 10, 2623 (2021).","journal-title":"Plants"},{"key":"30951_CR51","unstructured":"Harding, K. et al. Concepts in cryobionomics: a case study of Ribes genotype responses to cryopreservation in relation to thermal analysis oxidative stress nucleic acid methylation and transcriptional activity. In Cryopreservation of Crop Species in Europe, Proceedings of CRYOPLANET COST Action 871, 20\u201323 February 2008, Oulu, MTT Agrifood Research Working Papers 153 (eds Laamanen, J. et al.)."},{"key":"30951_CR52","doi-asserted-by":"crossref","unstructured":"A droplet vitrification cryopreservation protocol for conservation of hops (Humulus lupulus) genetic resources. Cryobiology 115, 104887 (2024).","DOI":"10.1016\/j.cryobiol.2024.104887"},{"key":"30951_CR53","doi-asserted-by":"publisher","first-page":"333","DOI":"10.1007\/s11627-013-9501-4","volume":"49","author":"ZF Yin","year":"2013","unstructured":"Yin, Z. F., Zhao, B., Bi, W. L., Chen, L. & Wang, Q. C. Direct shoot regeneration from basal leaf segments of lilium and assessment of genetic stability in regenerants by ISSR and AFLP markers. Vitro Cell. Develop Biol. - Plant. 49, 333\u2013342 (2013).","journal-title":"Vitro Cell. Develop Biol. - Plant."},{"key":"30951_CR54","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1007\/s11627-015-9704-y","volume":"51","author":"JM Zhang","year":"2015","unstructured":"Zhang, J. M. et al. Identification of a highly successful cryopreservation method (droplet-vitrification) for Petunia. Vitro Cell. Develop Biol. - Plant. 51, 445\u2013451 (2015).","journal-title":"Vitro Cell. Develop Biol. - Plant."},{"key":"30951_CR55","unstructured":"Reed, B. M. Plant Cryopreservation: A Practical Guide (Springer, 2005)."},{"key":"30951_CR56","doi-asserted-by":"publisher","first-page":"14674","DOI":"10.1038\/s41598-020-70869-3","volume":"10","author":"H Wilms","year":"2020","unstructured":"Wilms, H. et al. Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources. Sci. Rep. 10, 14674 (2020).","journal-title":"Sci. Rep."},{"key":"30951_CR57","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1007\/s11240-020-01841-2","volume":"144","author":"S Rantala","year":"2021","unstructured":"Rantala, S. et al. Droplet vitrification technique for cryopreservation of a large diversity of blackcurrant (Ribes nigrum L.) cultivars. Plant. Cell. Tiss Organ. Cult. 144, 79\u201390 (2021).","journal-title":"Plant. Cell. Tiss Organ. Cult."},{"key":"30951_CR58","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1007\/s11626-022-00680-z","volume":"58","author":"N Sharma","year":"2022","unstructured":"Sharma, N. et al. Cryopreservation and genetic stability assessment of regenerants of the critically endangered medicinal plant Dioscorea deltoidea Wall. Ex Griseb. For cryobanking of germplasm. Vitro Cell. Dev. Biol. -Plant. 58, 521\u2013529 (2022).","journal-title":"Vitro Cell. Dev. Biol. -Plant"},{"key":"30951_CR59","doi-asserted-by":"publisher","first-page":"1613","DOI":"10.1007\/s10722-016-0460-6","volume":"64","author":"GM Volk","year":"2017","unstructured":"Volk, G. M., Henk, A. D., Jenderek, M. M. & Richards, C. M. Probabilistic viability calculations for cryopreserving vegetatively propagated collections in genebanks. Genet. Resour. Crop Evol. 64, 1613\u20131622 (2017).","journal-title":"Genet. Resour. Crop Evol."},{"key":"30951_CR60","doi-asserted-by":"publisher","first-page":"473","DOI":"10.1111\/j.1399-3054.1962.tb08052.x","volume":"15","author":"T Murashige","year":"1962","unstructured":"Murashige, T. & Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473\u2013497 (1962).","journal-title":"Physiol. Plant."},{"key":"30951_CR61","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1007\/BF00232130","volume":"9","author":"A Sakai","year":"1990","unstructured":"Sakai, A., Kobayashi, S. & Oiyama, I. Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb. var. Brasiliensis Tanaka) by vitrification. Plant. Cell. Rep. 9, 30\u201333 (1990).","journal-title":"Plant. Cell. Rep."},{"key":"30951_CR62","first-page":"11","volume":"19","author":"J Doyle","year":"1987","unstructured":"Doyle, J. & Doyle, J. A rapid DNA isolation procedure from small quantities of fresh leaf tissues. Phytochem Bull. 19, 11\u201315 (1987).","journal-title":"Phytochem Bull."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30951-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30951-0","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30951-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T14:52:13Z","timestamp":1781621533000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30951-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,1]]},"references-count":62,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["30951"],"URL":"https:\/\/doi.org\/10.1038\/s41598-025-30951-0","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,1]]},"assertion":[{"value":"28 July 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"28 November 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 December 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"1356"}}