{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T05:03:18Z","timestamp":1787029398415,"version":"build-2736575974"},"reference-count":92,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T00:00:00Z","timestamp":1756771200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010801","name":"Xunta de Galicia (Spain)","doi-asserted-by":"publisher","award":["IN607A 2021\/06"],"award-info":[{"award-number":["IN607A 2021\/06"]}],"id":[{"id":"10.13039\/501100010801","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010801","name":"Xunta de Galicia (Spain)","doi-asserted-by":"publisher","award":["E-COST-GRANT-CA21157-8ad89b5c"],"award-info":[{"award-number":["E-COST-GRANT-CA21157-8ad89b5c"]}],"id":[{"id":"10.13039\/501100010801","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Autonomous Province of Trento","award":["IN607A 2021\/06"],"award-info":[{"award-number":["IN607A 2021\/06"]}]},{"name":"Autonomous Province of Trento","award":["E-COST-GRANT-CA21157-8ad89b5c"],"award-info":[{"award-number":["E-COST-GRANT-CA21157-8ad89b5c"]}]},{"name":"COST (European Cooperation in Science and Technology)","award":["IN607A 2021\/06"],"award-info":[{"award-number":["IN607A 2021\/06"]}]},{"name":"COST (European Cooperation in Science and Technology)","award":["E-COST-GRANT-CA21157-8ad89b5c"],"award-info":[{"award-number":["E-COST-GRANT-CA21157-8ad89b5c"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>This study aimed to investigate culture conditions for the efficient micropropagation of apple cultivars \u2018Golden Delicious\u2019 and \u2018Royal Gala\u2019 in liquid medium by temporary immersion. RITA\u00ae bioreactors were used for the multiplication stage whereas RITA\u00ae or Plantform\u2122 were used for the rooting stage. Murashige and Skoog media (MS) with N6-benzyladenine (BA) was used for shoot multiplication and indole-3-butyric acid (IBA) for root induction. During the multiplication phase, we evaluated the mineral medium, BA concentration, immersion frequency, silver nitrate and activated charcoal supplementation and the use of physical supports to hold explants in an upright position. The results demonstrated that longer incubation periods (10 weeks) were better than shorter periods (6 weeks) for decreasing hyperhydricity and increasing the multiplication coefficient (MC). For \u2018Golden Delicious\u2019, the highest MC were obtained either with explants placed directly on the bioreactor basket and immersed six times per day for 60 s in MS with 2.2 \u00b5M BA or explants placed between rockwool cubes cultivated with 4.4 \u00b5M BA (both yielding MC of 8.9 and 5\u201310% hyperhydricity). These results were superior to \u2018Royal Gala\u2019, which showed a MC of 7.3 and 23% of hyperhydricity when cultivated in MS with half nitrates, 1.55 \u00b5M BA and rockwool cubes. Both varieties rooted efficiently (96\u2013100%), and resulting plantlets were successfully acclimated. This is the first report in the micropropagation of these two commercial fruiting cultivars in temporary immersion, demonstrating the potential of this technology to enhance large-scale plant production for the apple nursery industry.<\/jats:p>","DOI":"10.3390\/plants14172740","type":"journal-article","created":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T12:04:28Z","timestamp":1756814668000},"page":"2740","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Micropropagation of Apple Cultivars \u2018Golden Delicious\u2019 and \u2018Royal Gala\u2019 in Bioreactors"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6517-4269","authenticated-orcid":false,"given":"Sim\u00f3n","family":"Miranda","sequence":"first","affiliation":[{"name":"Research and Innovation Centre, Fondazione Edmund Mach (FEM), Via Mach 1, 38098 San Michele all\u2019Adige, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5158-534X","authenticated-orcid":false,"given":"Mickael","family":"Malnoy","sequence":"additional","affiliation":[{"name":"Research and Innovation Centre, Fondazione Edmund Mach (FEM), Via Mach 1, 38098 San Michele all\u2019Adige, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anxela","family":"Aldrey","sequence":"additional","affiliation":[{"name":"Misi\u00f3n Biol\u00f3gica de Galicia, Consejo Superior de Investigaciones Cient\u00edficas, Avda de Vigo s\/n, 15705 Santiago de Compostela, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda Jos\u00e9","family":"Cernadas","sequence":"additional","affiliation":[{"name":"Misi\u00f3n Biol\u00f3gica de Galicia, Consejo Superior de Investigaciones Cient\u00edficas, Avda de Vigo s\/n, 15705 Santiago de Compostela, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1304-0736","authenticated-orcid":false,"given":"Conchi","family":"S\u00e1nchez","sequence":"additional","affiliation":[{"name":"Misi\u00f3n Biol\u00f3gica de Galicia, Consejo Superior de Investigaciones Cient\u00edficas, Avda de Vigo s\/n, 15705 Santiago de Compostela, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruce","family":"Christie","sequence":"additional","affiliation":[{"name":"The Greenplant Company, Palmerston North 4410, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8046-3133","authenticated-orcid":false,"given":"Nieves","family":"Vidal","sequence":"additional","affiliation":[{"name":"Misi\u00f3n Biol\u00f3gica de Galicia, Consejo Superior de Investigaciones Cient\u00edficas, Avda de Vigo s\/n, 15705 Santiago de Compostela, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Li, M., Xiao, Y., Mount, S., and Liu, Z. (2021). An Atlas of Genomic Resources for Studying Rosaceae Fruits and Ornamentals. Front. Plant Sci., 12.","DOI":"10.3389\/fpls.2021.644881"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Patocka, J., Bhardwaj, K., Klimova, B., Nepovimova, E., Wu, Q., Landi, M., Kuca, K., Valis, M., and Wu, W. (2020). Malus domestica: A Review on Nutritional Features, Chemical Composition, Traditional and Medicinal Value. Plants, 9.","DOI":"10.3390\/plants9111408"},{"key":"ref_3","unstructured":"(2025, July 28). Fruit Production by Variety Worldwide 2023. Available online: https:\/\/www.statista.com\/statistics\/264001\/worldwide-production-of-fruit-by-variety\/."},{"key":"ref_4","unstructured":"FAOSTAT (2025, July 28). Available online: https:\/\/www.fao.org\/faostat\/en\/#data\/QV\/visualize."},{"key":"ref_5","unstructured":"OECD (2019). Unclassified Environment Directorate Joint Meeting of the Chemicals Committee and the Working Party on Chemicals, Pesticides and Biotechnology. Consensus Document on the Biology of Apple (Malus domestica Borkh.), OECD. Available online: https:\/\/one.oecd.org\/document\/ENV\/JM\/MONO(2019)30\/En\/pdf."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1038\/s41588-020-00723-9","article-title":"Phased Diploid Genome Assemblies and Pan-Genomes Provide Insights into the Genetic History of Apple Domestication","volume":"52","author":"Sun","year":"2020","journal-title":"Nat. Genet."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1038\/ng.654","article-title":"The Genome of the Domesticated Apple (Malus \u00d7 domestica Borkh.)","volume":"42","author":"Velasco","year":"2010","journal-title":"Nat. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.1038\/ng.3886","article-title":"High-Quality de Novo Assembly of the Apple Genome and Methylome Dynamics of Early Fruit Development","volume":"49","author":"Daccord","year":"2017","journal-title":"Nat. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1007\/s00425-019-03100-x","article-title":"In Vitro Tissue Culture of Apple and Other Malus Species: Recent Advances and Applications","volume":"249","author":"Wang","year":"2019","journal-title":"Planta"},{"key":"ref_10","first-page":"1","article-title":"Plant Regeneration and Transformation in the Rosaceae","volume":"3","author":"Aldwinckle","year":"2009","journal-title":"Transgenic Plant J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1094\/PHYTO-07-23-0232-KC","article-title":"Application of Biotechniques for in Vitro Virus and Viroid Elimination in Pome Fruit crops","volume":"114","author":"Bettoni","year":"2024","journal-title":"Phytopathology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1186\/s13007-025-01344-1","article-title":"Production of HSVd- and PPV-free apricot cultivars by in vitro thermotherapy followed by meristem culture","volume":"21","author":"Burgos","year":"2025","journal-title":"Plant Methods"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2238","DOI":"10.4236\/ajps.2017.89150","article-title":"In Vitro Propagation and Recovery of Eight Apple and Two Pear Cultivars Held in a Germplasm Bank","volume":"8","author":"Fraga","year":"2017","journal-title":"Am. J. Plant Sci."},{"key":"ref_14","first-page":"965","article-title":"Micropropagation of the endangered species Malus niedzwetzkyana for conservation biodiversity in Kazakhstan","volume":"57","author":"Nurtaza","year":"2021","journal-title":"In Vitro Cell. Dev. Biol.-Plant"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Kom\u00e1rkov\u00e1, M., Cvr\u010dkov\u00e1, H., Dost\u00e1l, J., Buri\u00e1nek, V., and M\u00e1chov\u00e1, P. (2022). Long-Term Observation of In Vitro-Derived Malus sylvestris (L.) Mill., the Path from the Bud to the Tree. Apple Cultivation-Recent Advances, IntechOpen.","DOI":"10.5772\/intechopen.108359"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Abdalla, N., and Dobr\u00e1nszki, J. (2024). Meta-Topolin as an effective Benzyladenine derivative to improve the multiplication rate and quality of In Vitro axillary shoots of H\u00fasv\u00e9ti Rozmaring apple scion. Plants, 13.","DOI":"10.3390\/plants13111568"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1111\/pbi.13253","article-title":"Reduced Fire Blight Susceptibility in Apple Cultivars Using a High-efficiency CRISPR\/Cas9-FLP\/FRT-based Gene Editing System","volume":"18","author":"Pompili","year":"2020","journal-title":"Plant Biotechnol. J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1111\/tpj.16036","article-title":"Cas9 Genome-editing Applied to MdPGT1 in Apple Results in Reduced Foliar Phloridzin without Impacting Plant Growth","volume":"113","author":"Miranda","year":"2023","journal-title":"Plant J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"896","DOI":"10.1002\/elsc.201900041","article-title":"Use of Bioreactor Systems in the Propagation of Forest Trees","volume":"19","author":"Vidal","year":"2019","journal-title":"Eng. Life Sci."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Abd-Elsalam, K.A., and Prasad, R. (2018). Applications of Silver Nanoparticles in Plant Protection. Nanobiotechnology Applications in Plant Protection, Springer International Publishing.","DOI":"10.1007\/978-3-319-91161-8"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"P\u00e9rez-Caselles, C., Burgos, L., S\u00e1nchez-Balibrea, I., Egea, J.A., Faize, L., Mart\u00edn-Valmaseda, M., Bogdanchikova, N., Pestryakov, A., and Alburquerque, N. (2023). The effect of silver nanoparticle addition on micropropagation of apricot cultivars (Prunus armeniaca L.) in semisolid and liquid media. Plants, 12.","DOI":"10.3390\/plants12071547"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"110622","DOI":"10.1016\/j.scienta.2021.110622","article-title":"GreenTray\u00ae TIS bioreactor as an effective in vitro culture system for the micropropagation of Prunus spp. rootstocks and analysis of the plant-PGPMs interactions","volume":"291","author":"Cantabella","year":"2022","journal-title":"Sci. Hortic."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"494","DOI":"10.1007\/s11627-017-9856-z","article-title":"Temporary Immersion Systems for the Mass Propagation of Sweet Cherry Cultivars and Cherry Rootstocks: Development of a Micropropagation Procedure and Effect of Culture Conditions on Plant Quality","volume":"53","author":"Godoy","year":"2017","journal-title":"In Vitro Cell. Dev. Biol.-Plant"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Dolcet-Sanjuan, R., Casanovas, M., Franquesa, S., Alsina, E., Carrasco-Cuello, F., Torres, E., Rufat, J., Solsona, C., and Teixido, N. (2024). GreenTray\u00ae, a TIS Bioreactor for Plant Micropropagation and Abiotic or Biotic Stress Bioassays. Appl. Sci., 14.","DOI":"10.20944\/preprints202404.0793.v1"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Gago, D., S\u00e1nchez, C., Aldrey, A., Christie, C.B., Bernal, M.\u00c1., and Vidal, N. (2022). Micropropagation of Plum (Prunus domestica L.) in Bioreactors Using Photomixotrophic and Photoautotrophic Conditions. Horticulturae, 8.","DOI":"10.3390\/horticulturae8040286"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Gago, D., Bernal, M.\u00c1., S\u00e1nchez, C., Aldrey, A., Cuenca, B., Christie, C.B., and Vidal, N. (2022). Effect of Sucrose on Growth and Stress Status of Castanea sativa x C. crenata Shoots Cultured in Liquid Medium. Plants, 11.","DOI":"10.3390\/plants11070965"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1016\/j.biotechadv.2010.02.008","article-title":"Micropropagation of apple\u2014A review","volume":"28","year":"2010","journal-title":"Biotechnol. Adv."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1080\/14620316.2003.11511671","article-title":"Micropropagation of Apple Rootstock M.9 EMLA Using Bioreactor","volume":"78","author":"Chakrabarty","year":"2003","journal-title":"J. Hortic. Sci. Biotechnol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1007\/s11240-004-6659-9","article-title":"Optimisation of Growing Conditions for the Apple Rootstock M26 Grown in RITA Containers Using Temporary Immersion Principle","volume":"81","author":"Zhu","year":"2005","journal-title":"Plant Cell Tiss. Organ Cult."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Hvoslef-Eide, A.K., and Preil, W. (2005). Propagation of Prunus and Malus by temporary immersion. Liquid Culture Systems for In Vitro Plant Propagation, Springer.","DOI":"10.1007\/1-4020-3200-5"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1007\/s13580-020-00257-3","article-title":"Efficient production of virus-free apple plantlets using the temporary immersion bioreactor system","volume":"61","author":"Kim","year":"2020","journal-title":"Hortic. Environ. Biotechnol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"143","DOI":"10.17660\/ActaHortic.2009.817.13","article-title":"Process engineering as a means of regulating the microclimate in a photoautotrophic in vitro culture","volume":"817","author":"Fuljahn","year":"2009","journal-title":"Acta Hortic."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"12","DOI":"10.17221\/69\/2020-HORTSCI","article-title":"Evaluation of ElecTIS Bioreactor for the Micropropagation of Malus sylvestris (L.) Mill., an Important Autochthonous Species of Albania","volume":"48","author":"Sota","year":"2021","journal-title":"Hortic. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1007\/s11240-020-01935-x","article-title":"Effects of meta\u2013topolin derivatives and temporary immersion on hyperhydricity and in vitro shoot proliferation in Pyrus communis","volume":"143","author":"Lotfi","year":"2020","journal-title":"Plant Cell Tiss. Organ Cult."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1007\/s00468-017-1611-x","article-title":"Micropropagation of axillary shoots of Salix viminalis using a temporary immersion system","volume":"32","author":"Regueira","year":"2018","journal-title":"Trees"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"689","DOI":"10.4141\/cjps82-100","article-title":"Shoot tissue culture of apple: Comparative response of five cultivars to cytokinin and auxin","volume":"62","author":"Lane","year":"1982","journal-title":"Can. J. Plant Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1111\/j.1399-3054.1962.tb08052.x","article-title":"A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures","volume":"15","author":"Murashige","year":"1962","journal-title":"Physiol. Plant."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/j.scienta.2014.09.035","article-title":"Evaluation of a new vessel system based on temporary immersion system for micropropagation","volume":"179","author":"Welander","year":"2014","journal-title":"Sci. Hortic."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1007\/s11240-017-1285-5","article-title":"Micropropagation of axillary shoots of hybrid chestnut (Castanea sativa x C. crenata) in liquid medium in a continuous immersion system","volume":"131","author":"Cuenca","year":"2017","journal-title":"Plant Cell Tiss. Organ Cult."},{"key":"ref_40","unstructured":"(2025, July 30). New South Wales Government, Available online: https:\/\/www.dpi.nsw.gov.au\/agriculture\/horticulture\/pomes\/apples\/varieties\/gala."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1007\/BF00024677","article-title":"Micropropagation of thirteen Malus cultivars and rootstocks, and effect of antibiotics on proliferation","volume":"15","author":"Yepes","year":"1994","journal-title":"Plant Growth Reg."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1007\/s11240-004-4043-4","article-title":"Prevention of hyperhydricity in micropropagated carnation shoots by bottom cooling: Implications of oxidative stress","volume":"81","author":"Saher","year":"2005","journal-title":"Plant Cell Tiss. Org. Cult."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Polivanova, O.B., and Bedarev, V.A. (2022). Hyperhydricity in plant tissue culture. Plants, 11.","DOI":"10.3390\/plants11233313"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1007\/BF00029573","article-title":"The influence of cation and gelling agent concentrations on vitrification of apple cultivars in vitro","volume":"14","author":"Pasqualetto","year":"1988","journal-title":"Plant Cell Tiss. Org. Cult."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1007\/s11240-014-0568-3","article-title":"Effect of polyamines and polyamine precursors on hyperhydricity in micropropagated apple shoots","volume":"120","author":"Tabart","year":"2015","journal-title":"Plant Cell Tiss. Organ Cult."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1007\/s11240-023-02528-0","article-title":"Towards automated detection of hyperhydricity in plant in vitro culture","volume":"154","author":"Bethge","year":"2023","journal-title":"Plant Cell Tiss. Organ Cult."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1093\/treephys\/26.3.377","article-title":"Hyperhydricity in apple: Ultrastuctural and physiological aspects","volume":"26","author":"Chakrabarty","year":"2008","journal-title":"Tree Physiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1007\/s11240-015-0827-y","article-title":"A Temporary Immersion System for Micropropagation of Axillary Shoots of Hybrid Chestnut","volume":"123","author":"Vidal","year":"2015","journal-title":"Plant Cell Tiss. Organ Cult."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Nezami-Alanagh, E., Garoosi, G.A., Land\u00edn, M., and Gallego, P.P. (2019). Computer-based tools provide new insight into the key factors that cause physiological disorders of pistachio rootstocks cultured in vitro. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-46155-2"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1111\/j.1399-3054.1985.tb02379.x","article-title":"Anatomical and chemical studies on vitrified shoots of chestnut regenerated in vitro","volume":"65","author":"Vieitez","year":"1985","journal-title":"Physiol. Plant."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1007\/BF00037271","article-title":"Agar and ammonium nitrate influence hyperhydricity, tissue nitrate and total nitrogen content of serviceberry (Amelanchier arborea) shoots in vitro","volume":"35","author":"Brand","year":"1993","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1111\/j.1399-3054.1986.tb01239.x","article-title":"Ammonium-induced vitrification in cultured tissues","volume":"66","author":"Daguin","year":"1986","journal-title":"Physiol. Plant."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1023\/B:TICU.0000016825.18930.e4","article-title":"Hyperhydricity of micropropagated shoots: A typically stress-induced change of physiological state","volume":"77","author":"Kevers","year":"2004","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1080\/00103629909370193","article-title":"Foliar ammonium accumulation as an index of stress in plants","volume":"30","author":"Barker","year":"1999","journal-title":"Commun. Soil Sci. Plant Anal."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1413","DOI":"10.1093\/jxb\/ert019","article-title":"Ammonium-induced shoot ethylene production is associated with the inhibition of lateral root formation in Arabidopsis","volume":"64","author":"Li","year":"2013","journal-title":"J. Exp. Bot."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1006\/anbo.2000.1299","article-title":"Micropropagation of potato; evaluation of closed, diffusive and forced ventilation on growth and tuberization","volume":"87","author":"Zobayed","year":"2001","journal-title":"Ann. Bot."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1023\/A:1015668610465","article-title":"Temporary immersion systems in plant micropropagation","volume":"69","author":"Etienne","year":"2002","journal-title":"Plant Cell Tiss. Organ Cult."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Hvoslef-Eide, A.K., and Preil, W. (2005). Aeration stress in plant tissue cultures. Liquid Culture Systems for In Vitro Plant Propagation, Springer.","DOI":"10.1007\/1-4020-3200-5"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1104\/pp.112.202853","article-title":"Proteome analysis in arabidopsis reveals shoot- and root-specific targets of cytokinin action and differential regulation of hormonal homeostasis","volume":"161","author":"Malbeck","year":"2013","journal-title":"Plant Physiol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/j.scienta.2017.02.010","article-title":"Effects of multiple factors on hyperhydricity of Allium sativum L.","volume":"217","author":"Liu","year":"2017","journal-title":"Sci. Hortic."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1016\/S0981-9428(01)01309-2","article-title":"Transport and distribution of benzyladenine in Actinidia deliciosa explants cultured in liquid and solid media","volume":"39","author":"Feito","year":"2001","journal-title":"Plant Physiol. Biochem."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1007\/s11240-011-0088-3","article-title":"Morphological and physiological responses of proliferating shoots of teak to temporary immersion and BA treatments","volume":"109","author":"Quiala","year":"2012","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Sreelekshmi, R., and Siril, E.A. (2021). Effective reversal of hyperhydricity leading to efficient micropropagation of Dianthus chinensis L.. 3Biotech, 11.","DOI":"10.1007\/s13205-021-02645-7"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1007\/s11240-020-01937-9","article-title":"Temporary immersion systems to improve alder micropropagation","volume":"143","author":"Cernadas","year":"2020","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Chmielarz, P., S\u00e1nchez, C., Martins, J.R., Ley-L\u00f3pez, J.M., Covelo, P., Cernadas, M.J., Aldrey, A., Rico, S., Vielba, J.M., and Christie, B. (2025). Multiplication of axillary shoots of adult Quercus robur L. trees in RITA\u00ae bioreactors. Forests, 16.","DOI":"10.3390\/f16081285"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1080\/11263504.2017.1340356","article-title":"The effect of Plantform\u2122 bioreactor on micropropagation of Quercus robur in comparison to a conventional in vitro culture system on gelled medium, and assessment of the microenvironment influence on leaf structure","volume":"151","author":"Gatti","year":"2017","journal-title":"Plant Biosyst."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.plantsci.2016.10.008","article-title":"Reversion of hyperhydricity in pink (Dianthus chinensis L.) plantlets by AgNO3 and its associated mechanism during in vitro culture","volume":"254","author":"Gao","year":"2017","journal-title":"Plant Sci."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"111593","DOI":"10.1016\/j.scienta.2022.111593","article-title":"Silver nitrate reduces hyperhydricity in shoots regenerated from the hypocotyl of snapdragon cv. Maryland Apple Blossom","volume":"308","author":"Lee","year":"2023","journal-title":"Sci. Hortic."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"1311","DOI":"10.1007\/s12298-019-00689-x","article-title":"The effect of silver nitrate on micropropagation of Moringa oleifera Lam. an important vegetable crop of tropics with substantial nutritional value","volume":"25","author":"Siril","year":"2019","journal-title":"Physiol. Mol. Biol. Plants"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"8","DOI":"10.2225\/vol12-issue2-fulltext-1","article-title":"AgNO3\u2014A potential regulator of ethylene activity and plant growth modulator","volume":"12","author":"Kumar","year":"2009","journal-title":"Electron. J. Biotech."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"55","DOI":"10.15446\/rev.colomb.biote.v15n2.41265","article-title":"Effect of culture medium consistence and silver nitrate on micropropagation of two potato (Solanum tuberosum) cultivars","volume":"15","author":"Ticona","year":"2013","journal-title":"Rev. Colomb. Biotecnol."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1111\/j.1399-3054.1994.tb02207.x","article-title":"The effect of the gaseous state on bud induction and shoot multiplication in vitro in eastern white cedar","volume":"90","author":"Nour","year":"1994","journal-title":"Physiol. Plant."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1023\/A:1005751017498","article-title":"Ethylene involvement in in vitro organogenesis and plant growth of Populus tremula L.","volume":"22","author":"Arigita","year":"1997","journal-title":"Plant Growth Regul."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1007\/BF01690260","article-title":"Ethylene enhances shoot formation in cultures of the peach rootstock GF-677 (Prunus persica \u00d7 P. amygdalus)","volume":"15","author":"Economou","year":"1995","journal-title":"Plant Cell Rep."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"3585","DOI":"10.1105\/tpc.108.065185","article-title":"Silver ions increase auxin efflux independently of effects on ethylene response","volume":"21","author":"Strader","year":"2009","journal-title":"Plant Cell"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1023\/A:1006119015972","article-title":"The use of charcoal in in vitro culture\u2014A review","volume":"26","author":"Pan","year":"1998","journal-title":"Plant Growth Reg."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1016\/j.biotechadv.2008.08.003","article-title":"The role of activated charcoal in plant tissue culture","volume":"26","author":"Thomas","year":"2008","journal-title":"Biotechnol. Adv."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"103","DOI":"10.30574\/wjarr.2020.6.2.0137","article-title":"Effects of culture medium strength and antioxidants on adventitious bud multiplication, hyperhydricity and tissue browning of date palm cv. Aziza Bouzid","volume":"6","author":"Elmaataoui","year":"2020","journal-title":"World J. Adv. Res. Rev."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1023\/A:1006471519609","article-title":"Effect of activated charcoal and 6-benzyladenine on in vitro nitrogen uptake by Lagerstroemia indica","volume":"63","author":"Eymar","year":"2000","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/0304-4238(93)90082-2","article-title":"Involvement of activated charcoal in the modulation of abiotic and biotic ethylene levels in tissue cultures","volume":"54","author":"Panizza","year":"1993","journal-title":"Sci. Hort."},{"key":"ref_81","first-page":"98","article-title":"Effects of activated charcoal on rooting of in vitro apple (Malus domestica Borkh.) shoots","volume":"7","author":"Szalai","year":"2001","journal-title":"Int. J. Hortic. Sci."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1023\/A:1000309023415","article-title":"Optimization of culture media for in vitro rooting of Malus domestica Borkh. cv. Compact Spartan","volume":"39","author":"Druart","year":"1997","journal-title":"Biol. Plant."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/S0304-4238(98)00259-3","article-title":"Micropropagation of apple cv. Tydeman Early Worcester","volume":"81","author":"Modgil","year":"1999","journal-title":"Sci. Hortic."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"209","DOI":"10.17660\/ActaHortic.2009.839.25","article-title":"Commercially feasible protocol for rooting and acclimatization of micropropagated apple rootstocks","volume":"839","author":"Modgil","year":"2009","journal-title":"Acta Hortic."},{"key":"ref_85","first-page":"26","article-title":"Posteffects of cytokinins and auxin levels of proliferation media on rooting ability of in vitro apple shoots (Malus domestica Borkh.) \u2018Red Fuji\u2019","volume":"7","author":"Szalai","year":"2001","journal-title":"Int. J. Hortic. Sci."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1023\/A:1025072922054","article-title":"The influence of medium aeration on in vitro rooting of Australian plant microcuttings","volume":"75","author":"Newel","year":"2003","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_87","unstructured":"Teixeira da Silva, J. (2006). Matrix-Supported Liquid Culture Systems for Efficient Micropropagation of Floricultural Plants. Floriculture, Ornamental and Plant Biotechnology, Global Science Books."},{"key":"ref_88","first-page":"16","article-title":"Polymerized peat plugs improve American chestnut somatic embryo germination in vitro","volume":"24","author":"Maner","year":"2010","journal-title":"J. Am. Chestnut Found."},{"key":"ref_89","doi-asserted-by":"crossref","unstructured":"Liu, Z., Bi, W.-L., Shukla, M.R., and Saxena, P.K. (2022). In Vitro Technologies for American Chestnut (Castanea dentata (Marshall) Borkh) Conservation. Plants, 11.","DOI":"10.3390\/plants11030464"},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1007\/s11627-020-10136-2","article-title":"In vitro co-culture system for Prunus spp. and Armillaria mellea in phenolic foam rooting matric","volume":"57","author":"Adelberg","year":"2021","journal-title":"In Vitro Cell. Dev. Biol.-Plant"},{"key":"ref_91","unstructured":"Terzi, M., Cella, R., and Falavigna, A. (1995). A new concept of plant in vitro cultivation liquid medium: Temporary immersion. Current Issues in Plant Molecular and Cellular Biology. In Current Plant Science and Biotechnology in Agriculture, Kluver Academic Publishers."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1007\/978-1-0716-3654-1_16","article-title":"Proliferation of Axillary Shoots of Chestnut in Temporary Immersion Systems","volume":"Volume 2759","year":"2024","journal-title":"Micropropagation Methods in Temporary Immersion Systems"}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/14\/17\/2740\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:38:01Z","timestamp":1760035081000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/14\/17\/2740"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,2]]},"references-count":92,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["plants14172740"],"URL":"https:\/\/doi.org\/10.3390\/plants14172740","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,2]]}}}