{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T18:49:01Z","timestamp":1788374941488,"version":"build-2803163510"},"reference-count":49,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,3,30]],"date-time":"2026-03-30T00:00:00Z","timestamp":1774828800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Temporary Immersion Systems (TISs) are an efficient alternative for in vitro plant regeneration. This study aimed to evaluate the effect of different culture methods on the in vitro shoot proliferation and acclimatization of agave (Agave marmorata Roezl). The culture methods compared were a recipient for automated temporary immersion (RITA\u00ae), a temporary immersion bioreactor (TIB), a SETIS\u2122 bioreactor, and a semisolid medium control. After eight weeks of in vitro culture, the hyperhydricity of the explants, development variables, photosynthetic pigment content, stomatal density, and survival percentage during acclimatization were evaluated. The results showed that TISs significantly reduced explant hyperhydricity and increased the multiplication rate, number of shoots and leaves, number of roots per shoot, root length, carotenoid content, stomatal density, and percentage of closed stomata during in vitro shoot proliferation. Furthermore, TISs resulted in a higher number of leaves and roots and improved the survival percentage during acclimatization compared to the semisolid medium. Explants cultured in the SETIS\u2122 bioreactor showed the highest photosynthetic pigment content. In conclusion, the evaluated TISs enhanced the physiological development of the explants, favoring the multiplication rate and survival percentage during the acclimatization of A. marmorata.<\/jats:p>","DOI":"10.3390\/plants15071054","type":"journal-article","created":{"date-parts":[[2026,3,30]],"date-time":"2026-03-30T09:57:11Z","timestamp":1774864631000},"page":"1054","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Evaluation of Different Bioreactors During In Vitro Shoot Proliferation and Acclimatization of Agave (Agave marmorata Roezl)"],"prefix":"10.3390","volume":"15","author":[{"given":"Mar\u00eda del Rosario","family":"Moreno-Hern\u00e1ndez","sequence":"first","affiliation":[{"name":"Colegio de Postgraduados Campus C\u00f3rdoba, Amatl\u00e1n de los Reyes 94953, Veracruz, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eucario","family":"Mancilla-\u00c1lvarez","sequence":"additional","affiliation":[{"name":"Colegio de Postgraduados Campus C\u00f3rdoba, Amatl\u00e1n de los Reyes 94953, Veracruz, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Abel","family":"L\u00f3pez-Buenfil","sequence":"additional","affiliation":[{"name":"Colegio de Postgraduados Campus Montecillo, Texcoco 56264, Estado de M\u00e9xico, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2617-3079","authenticated-orcid":false,"given":"Jeric\u00f3 Jab\u00edn","family":"Bello-Bello","sequence":"additional","affiliation":[{"name":"Secretar\u00eda de Ciencia, Humanidades, Tecnolog\u00eda e Innovaci\u00f3n, Colegio de Postgraduados Campus C\u00f3rdoba, Amatl\u00e1n de los Reyes 94953, Veracruz, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1381","DOI":"10.1002\/ppp3.10532","article-title":"Evolution history and the importance of genomic diversity facing climate change. The case of Agave marmorata Roezl., a microendemic agave used for mezcal production","volume":"6","author":"Klimova","year":"2024","journal-title":"Plants People Planet"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Ruiz-Mondrag\u00f3n, K.Y., and Eguiarte, L.E. (2025). Plant Breeding and Potential Biotechnology Uses of Agave as a Source of Fiber, with Emphasis on Maguey Tepeztate (Agave marmorata Roezl) From Central and South Mexico. Breeding and Biotechnology of Leaf, Fruit, and Seed Fiber Crops, Springer Nature.","DOI":"10.1007\/978-3-032-00407-9_2"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Martinez-Rodriguez, A., Beltran-Garc\u00eda, C., Valdez-Salas, B., Santacruz-Ruvalcaba, F., Di Mascio, P., and Beltran-Garc\u00eda, M.J. (2022). Micropropagation of seed-derived clonal lines of the endangered Agave marmorata Roezl and their compatibility with endophytes. Biology, 11.","DOI":"10.3390\/biology11101423"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1007\/s12231-010-9117-0","article-title":"Population dynamics of Agave marmorata Roezl. under two contrasting management systems in central Mexico","volume":"64","author":"Caballero","year":"2010","journal-title":"Econ. Bot."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/j.sajb.2024.04.011","article-title":"Temporary immersion bioreactor as an efficient method for in vitro propagation of Agave marmorata","volume":"169","year":"2024","journal-title":"S. Afr. J. Bot."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1007\/s11627-023-10362-4","article-title":"Micropropagaci\u00f3n de palma de iraca (Carludovica palmata Ruiz y Pav) mediante un sistema de inmersi\u00f3n temporal","volume":"59","year":"2023","journal-title":"In Vitro Cell. Dev. Biol.\u2014Plant"},{"key":"ref_7","first-page":"1003","article-title":"In vitro propagation of Agave guiengola Gentry using semisolid medium and temporary immersion bioreactors","volume":"90","year":"2021","journal-title":"Phyton\u2014Int. J. Exp. Bot."},{"key":"ref_8","first-page":"493","article-title":"Regeneraci\u00f3n in vitro de Agave wocomahi Gentry (Asparagaceae)","volume":"45","author":"Ocampo","year":"2022","journal-title":"Rev. Fitotec. Mex."},{"key":"ref_9","first-page":"139","article-title":"Micropropagation of Agave maximiliana baker by axillary shoot proliferation","volume":"54","author":"Portillo","year":"2022","journal-title":"Polibot\u00e1nica"},{"key":"ref_10","first-page":"83","article-title":"In vitro-propagation of Agave tequilana weber cv. azul in a temporary immersion system","volume":"91","author":"Balch","year":"2022","journal-title":"Phyton\u2014Int. J. Exp. Bot."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"883","DOI":"10.17129\/botsci.3279","article-title":"In vitro culture of Agave potatorum a threatened species, endemic to Mexico","volume":"101","year":"2023","journal-title":"Bot. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Seliem, M.K., Abdalla, N., and El-Mahrouk, M.E. (2025). Cytokinin potentials on In Vitro shoot proliferation and subsequent rooting of Agave sisalana Perr. Syn. Horticulturae, 11.","DOI":"10.3390\/horticulturae11080929"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1007\/s11627-024-10480-7","article-title":"Scaling-up procedures and factors for mass micropropagation using temporary immersion systems","volume":"61","year":"2025","journal-title":"In Vitro Cell. Dev. Biol.\u2014Plant"},{"key":"ref_14","first-page":"77","article-title":"Comparison of conventional and temporary immersion systems on micropropagation (multiplication phase) of Agave angustifolia Haw","volume":"11","author":"Robert","year":"2021","journal-title":"Bacanora. 3 Biotech"},{"key":"ref_15","first-page":"2694","article-title":"Mass propagation of tobala mezcal maguey (Agave potatorum Zucc.) in a temporary immersion system compared with a solid medium","volume":"16","year":"2023","journal-title":"Agro Product."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Ram\u00edrez-Mosqueda, M.A., and Cruz-Cruz, C.A. (2024). BioMINT: A Temporary Immersion System for Agave Micropropagation. Micropropagation Methods in Temporary Immersion Systems, Humana.","DOI":"10.1007\/978-1-0716-3654-1"},{"key":"ref_17","first-page":"69","article-title":"In Vitro Multiplication of Agave (A. marmorata and A. potatorum) by Temporary Immersion in SETIS\u2122 Bioreactor","volume":"Volume 2759","year":"2024","journal-title":"Micropropagation Methods in Temporary Immersion Systems; Methods in Molecular Biology"},{"key":"ref_18","first-page":"74","article-title":"Temporary immersion systems induce photomixotrophism during in vitro propagation of agave Tobal\u00e1","volume":"14","year":"2024","journal-title":"3 Biotech"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1007\/BF00040116","article-title":"Comparison of methods of liquid medium culture for banana micropropagation","volume":"32","author":"Alvard","year":"1993","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1007\/s002990050653","article-title":"Pineapple (Ananas comosus L. Merr) micropropagation in temporary immersion systems","volume":"18","author":"Escalona","year":"1999","journal-title":"Plant Cell Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1016\/j.sajb.2022.03.009","article-title":"In vitro multiplication of agave tobala (Agave potatorum Zucc.) using Ebb-and-Flow bioreactor","volume":"147","year":"2022","journal-title":"S. Afr. J. Bot."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1007\/s11240-018-1508-4","article-title":"Comparison of two different micropropagation systems of Saccharum officinarum L. and expression analysis of PIP2; 1 and EIN3 genes as efficiency system indicators","volume":"136","author":"Robert","year":"2019","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_23","unstructured":"(2025, November 10). SETIS\u2122 Bioreactor Temporary Immersion Systems in Plant Micropropagation. Available online: https:\/\/setis-systems.be\/home."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1016\/j.sajb.2020.08.019","article-title":"Light-emitting diodes and gas exchange facilitation minimize hyperhydricity in Lippia grata: Physiological, biochemical and morpho anatomical aspects","volume":"135","author":"Neto","year":"2020","journal-title":"S. Afr. J. Bot."},{"key":"ref_25","first-page":"84","article-title":"Regeneration of Agave marmorata Roelz plants in temporary immersion systems, via organogenesis and somatic embryogenesis","volume":"24","year":"2021","journal-title":"Trop. Subtrop. Agroecosyst."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2366","DOI":"10.1111\/fwb.12216","article-title":"Optimising hydrological conditions to sustain wintering waterbird populations in Poyang Lake National Natural Reserve: Implications for dam operations","volume":"58","author":"Wang","year":"2013","journal-title":"Freshw. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1007\/s11627-019-09973-7","article-title":"A new temporary immersion system for commercial micropropagation of banana (Musa AAA cv. Grand Naine)","volume":"55","year":"2019","journal-title":"In Vitro Cell. Dev. Biol.\u2014Plant"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Gross, J. (1991). Carotenoids. Pigments in Vegetables, Chlorophylls and Carotenoids, Springer.","DOI":"10.1007\/978-1-4615-2033-7"},{"key":"ref_29","unstructured":"Watson, R.R. (2019). Phenotypic diversity of colored phytochemicals in sorghum accessions with various pericarp pigments. Polyphenols in Plants: Isolation, Purification and Extract Preparation, Elsevier."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"100203","DOI":"10.1016\/j.cpb.2021.100203","article-title":"Vital roles of carotenoids in plants and humans to deteriorate stress with its structure, biosynthesis, metabolic engineering and functional aspects","volume":"26","author":"Swapnil","year":"2021","journal-title":"Curr. Plant Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1007\/s00425-022-03953-9","article-title":"Biotechnological production of \u03b2-carotene using plant in vitro cultures","volume":"256","author":"Almagro","year":"2022","journal-title":"Planta"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Nguyen, T.N.P., and Sung, J. (2025). Light Spectral-Ranged Specific Metabolisms of Plant Pigments. Metabolites, 15.","DOI":"10.3390\/metabo15010001"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Hwang, H.D., Kwon, S.H., Murthy, H.N., Yun, S.W., Pyo, S.S., and Park, S.Y. (2022). Temporary immersion bioreactor system as an efficient method for mass production of in vitro plants in horticulture and medicinal plants. Agronomy, 12.","DOI":"10.3390\/agronomy12020346"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"20371","DOI":"10.1038\/s41598-021-99923-4","article-title":"A novel temporary immersion bio-reactor system for large scale multiplication of banana (Rasthali AAB\u2014Silk)","volume":"11","author":"Uma","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Lim, M.J., Han, J.E., Murthy, H.N., Song, H.Y., Lee, S.Y., and Park, S.Y. (2024). A comparison of semisolid, liquid, and temporary immersion bioreactor systems for effective plant regeneration of Gerbera jamesonii \u201cShy Pink\u201d. Horticulturae, 10.","DOI":"10.21203\/rs.3.rs-4366227\/v1"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2200116","DOI":"10.1002\/adbi.202200116","article-title":"Minor changes in daily rhythms induced by a skeleton photoperiod are associated with increased adiposity and glucose intolerance","volume":"7","author":"Rumanova","year":"2023","journal-title":"Adv. Biol."},{"key":"ref_37","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_38","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1007\/s11240-022-02441-y","article-title":"Micropropagation of Brassavola nodosa (L.) Lindl. using SETIS\u2122 bioreactor","volume":"153","author":"Vendrame","year":"2023","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.bbrc.2022.06.097","article-title":"Root-specific activation of plasma membrane H+-ATPase 1 enhances plant growth and shoot accumulation of nutrient elements under nutrient-poor conditions in Arabidopsis thaliana","volume":"621","author":"Monden","year":"2022","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1016\/j.plaphy.2021.12.019","article-title":"Light regulation of potassium in plants","volume":"170","author":"Ahammed","year":"2022","journal-title":"Plant Physiol. Biochem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"976","DOI":"10.1111\/nph.14332","article-title":"Photosynthetic responses to altitude: An explanation based on optimality principles","volume":"213","author":"Wang","year":"2017","journal-title":"New Phytol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.1007\/s11627-022-10314-4","article-title":"In vitro germination and development of \u201cCanelita\u201d (Lycaste aromatica (Graham) Lindl.) in gravity immersion bioreactors","volume":"58","year":"2022","journal-title":"In Vitro Cell. Dev. Biol.\u2014Plant"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1111\/j.1365-3040.2005.01319.x","article-title":"Stomatal density, anatomy and nutrient concentrations of Scots pine needles are affected by elevated CO2 and temperature","volume":"28","author":"Luomala","year":"2005","journal-title":"Plant Cell Environ."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1007\/s11627-025-10527-3","article-title":"Comparative structural analysis of banana plantlets cultivated in semisolid medium and in temporary immersion bioreactor","volume":"61","author":"Luis","year":"2025","journal-title":"In Vitro Cell. Dev. Biol.\u2014Plant"},{"key":"ref_45","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_46","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1007\/s10725-012-9702-6","article-title":"Effect of methyl jasmonate treatments on alleviation of polyethylene glycol-mediated water stress in banana (Musa acuminata cv. \u2018Berangan\u2019, AAA) shoot tip cultures","volume":"68","author":"Mahmood","year":"2012","journal-title":"Plant Growth Regul."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1111\/j.1750-3841.2009.01417.x","article-title":"Comparison of 3 spectrophotometric methods for carotenoid determination in frequently consumed fruits and vegetables","volume":"75","author":"Biehler","year":"2010","journal-title":"J. Food Sci."},{"key":"ref_48","first-page":"1210","article-title":"In vitro free radical scavenging activity of wild edible mushroom, Pleurotus squarrosulus (Mont.) singer","volume":"48","author":"Pal","year":"2010","journal-title":"J. Exp. Biol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"3317","DOI":"10.1093\/jxb\/ern185","article-title":"Responses of leaf stomatal density to water status and its relationship with photosynthesis in a grass","volume":"59","author":"Xu","year":"2008","journal-title":"J. Exp. Bot."}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/7\/1054\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,30]],"date-time":"2026-03-30T10:04:29Z","timestamp":1774865069000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/7\/1054"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,30]]},"references-count":49,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["plants15071054"],"URL":"https:\/\/doi.org\/10.3390\/plants15071054","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,30]]}}}