{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T11:02:33Z","timestamp":1771930953734,"version":"3.50.1"},"reference-count":62,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,1,2]],"date-time":"2023-01-02T00:00:00Z","timestamp":1672617600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["SFRH\/BD\/146485\/2019"],"award-info":[{"award-number":["SFRH\/BD\/146485\/2019"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["UIDB\/04004\/2020"],"award-info":[{"award-number":["UIDB\/04004\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"name":"University of Coimbra\/Santander Seed Projects Award 2021","award":["SFRH\/BD\/146485\/2019"],"award-info":[{"award-number":["SFRH\/BD\/146485\/2019"]}]},{"name":"University of Coimbra\/Santander Seed Projects Award 2021","award":["UIDB\/04004\/2020"],"award-info":[{"award-number":["UIDB\/04004\/2020"]}]},{"DOI":"10.13039\/501100001871","name":"Center for Functional Ecology\u2014Science for People and Planet (CFE)","doi-asserted-by":"publisher","award":["SFRH\/BD\/146485\/2019"],"award-info":[{"award-number":["SFRH\/BD\/146485\/2019"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Center for Functional Ecology\u2014Science for People and Planet (CFE)","doi-asserted-by":"publisher","award":["UIDB\/04004\/2020"],"award-info":[{"award-number":["UIDB\/04004\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Plant cell suspension cultures are widely used as a tool for analyzing cellular and molecular processes, metabolite synthesis, and differentiation, bypassing the structural complexity of plants. Within the range of approaches used to increase the production of metabolites by plant cells, one of the most recurrent is applying elicitors capable of stimulating metabolic pathways related to defense mechanisms. Previous proteomics analysis of tamarillo cell lines and cell suspension cultures have been used to further characterize and optimize the growth and stress-related metabolite production under in vitro controlled conditions. The main objective of this work was to develop a novel plant-based bioreactor system to produce hydrolytic enzymes using an elicitation approach. Based on effective protocols for tamarillo micropropagation and plant cell suspension culture establishment from induced callus lines, cell growth has been optimized, and enzymatic activity profiles under in vitro controlled conditions characterized. By testing different sucrose concentrations and the effects of two types of biotic elicitors, it was found that 3% (w\/v) sucrose concentration in the liquid medium enhanced the production of hydrolytic enzymes. Moreover, casein hydrolysate at 0.5 and 1.5 g\/L promoted protein production, whereas yeast extract (0.5 g\/L) enhanced glycosidase activity. Meanwhile, chitosan (0.05 and 0.1 g\/L) enhanced glycosidases, alkaline phosphates, and protease activities.<\/jats:p>","DOI":"10.3390\/plants12010190","type":"journal-article","created":{"date-parts":[[2023,1,3]],"date-time":"2023-01-03T02:05:53Z","timestamp":1672711553000},"page":"190","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Enhancing the Production of Hydrolytic Enzymes in Elicited Tamarillo (Solanum betaceum Cav.) Cell Suspension Cultures"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1224-3056","authenticated-orcid":false,"given":"Bruno","family":"Casimiro","sequence":"first","affiliation":[{"name":"Centre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, Cal\u00e7ada Martim de Freitas, University of Coimbra, 3000-456 Coimbra, Portugal"}]},{"given":"In\u00eas","family":"Mota","sequence":"additional","affiliation":[{"name":"Centre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, Cal\u00e7ada Martim de Freitas, University of Coimbra, 3000-456 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0532-4242","authenticated-orcid":false,"given":"Paula","family":"Ver\u00edssimo","sequence":"additional","affiliation":[{"name":"Centre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, Cal\u00e7ada Martim de Freitas, University of Coimbra, 3000-456 Coimbra, Portugal"},{"name":"Center for Neuroscience and Cell Biology, University of Coimbra, Cal\u00e7ada Martim de Freitas, 3000-456 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2299-298X","authenticated-orcid":false,"given":"Jorge","family":"Canhoto","sequence":"additional","affiliation":[{"name":"Centre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, Cal\u00e7ada Martim de Freitas, University of Coimbra, 3000-456 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2151-3916","authenticated-orcid":false,"given":"Sandra","family":"Correia","sequence":"additional","affiliation":[{"name":"Centre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, Cal\u00e7ada Martim de Freitas, University of Coimbra, 3000-456 Coimbra, Portugal"},{"name":"InnovPlantProtect CoLab, Estrada de Gil Vaz, 7351-901 Elvas, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Moon, K.-B., Park, J.-S., Park, Y.-I., Song, I.-J., Lee, H.-J., Cho, H.S., Jeon, J.-H., and Kim, H.-S. (2019). Development of systems for the production of plant-derived biopharmaceuticals. Plants, 9.","DOI":"10.3390\/plants9010030"},{"key":"ref_2","first-page":"1573","article-title":"The emergency response capacity of plant-based biopharmaceutical manufacturing-what it is and what it could be","volume":"11","author":"Nandi","year":"2020","journal-title":"Front. Plant Sci."},{"key":"ref_3","first-page":"1","article-title":"Rapid Expression of COVID-19 proteins by transient expression in tobacco. Author Contributions","volume":"1","author":"Lindsay","year":"2020","journal-title":"bioRxiv"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1415","DOI":"10.1038\/nbt1027","article-title":"Plant cell cultures for the production of recombinant proteins","volume":"22","author":"Hellwig","year":"2004","journal-title":"Nat. Biotechnol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"278","DOI":"10.1016\/j.biotechadv.2011.01.002","article-title":"Towards high-yield production of pharmaceutical proteins with plant cell suspension cultures","volume":"29","author":"Xu","year":"2011","journal-title":"Biotechnol. Adv."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5503","DOI":"10.2174\/1381612811319310006","article-title":"N-glycosylation of plant-produced recombinant proteins","volume":"19","author":"Bosch","year":"2013","journal-title":"Curr. Pharm. Des."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"297","DOI":"10.3389\/fpls.2016.00297","article-title":"Putting the spotlight back on plant suspension cultures","volume":"7","author":"Santos","year":"2016","journal-title":"Front. Plant Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1111\/pbi.12147","article-title":"Scale-up of hydrophobin-assisted recombinant protein production in tobacco BY-2 suspension cells","volume":"12","author":"Reuter","year":"2014","journal-title":"Plant Biotechnol. J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"119501","DOI":"10.1016\/j.saa.2021.119501","article-title":"Nutraceutical properties of tamarillo fruits: A vibrational study","volume":"252","author":"Martin","year":"2021","journal-title":"Spectrochim. Acta-Part A Mol. Biomol. Spectrosc."},{"key":"ref_10","unstructured":"Jain, S., and Gupta, P. (2018). Somatic Embryogenesis of Tamarillo (Solanum betaceum Cav.), Springer."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1666","DOI":"10.1021\/pr200856w","article-title":"Comparative proteomic analysis of auxin-induced embryogenic and nonembryogenic tissues of the solanaceous tree Chyphomandra betacea (Tamarillo)","volume":"11","author":"Correia","year":"2012","journal-title":"J. Proteome Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"438","DOI":"10.3389\/fpls.2019.00438","article-title":"NEP-TC a rRNA methyltransferase involved on somatic embryogenesis of tamarillo (Solanum betaceum Cav.)","volume":"10","author":"Correia","year":"2019","journal-title":"Front. Plant Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"606","DOI":"10.1007\/s11627-017-9864-z","article-title":"Establishment and biochemical characterization of tamarillo (Solanum betaceum Cav.) embryogenic cell suspension cultures","volume":"53","author":"Alves","year":"2017","journal-title":"Vitr. Cell. Dev. Biol.-Plant"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"80062","DOI":"10.1007\/s11356-022-20938-z","article-title":"\u03b2-glucan and its nanocomposites in sustainable agriculture and environment: An overview of mechanisms and applications","volume":"29","author":"Chavanke","year":"2022","journal-title":"Environ. Sci. Pollut. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1983","DOI":"10.1007\/s10529-012-1012-6","article-title":"Plant \u03b2-1,3-glucanases: Their biological functions and transgenic expression against phytopathogenic fungi","volume":"34","author":"Balasubramanian","year":"2012","journal-title":"Biotechnol. Lett."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Caseiro, C., Dias, J.N.R., de Andrade Fontes, C.M.G., and Bule, P. (2022). From cancer therapy to winemaking: The molecular structure and applications of \u03b2-glucans and \u03b2-1, 3-glucanases. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23063156"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1007\/978-3-319-48006-0_6","article-title":"Control of fungal diseases in agricultural crops by chitinase and glucanase transgenes","volume":"22","author":"Sandhu","year":"2017","journal-title":"Sustainable Agriculture Reviews"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"21","DOI":"10.4103\/0975-7406.106559","article-title":"Chitinases: An update","volume":"5","author":"Javed","year":"2013","journal-title":"J. Pharm. Bioallied Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1007\/s11738-006-0017-y","article-title":"Stress-induced expression of cucumber chitinase and Nicotiana plumbaginifolia \u03b2-1,3-glucanase genes in transgenic potato plants","volume":"29","author":"Salaj","year":"2007","journal-title":"Acta Physiol. Plant."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"7957269","DOI":"10.1155\/2018\/7957269","article-title":"The role of microbial aspartic protease enzyme in food and beverage industries","volume":"2018","author":"Mamo","year":"2018","journal-title":"J. Food Qual."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1227","DOI":"10.1007\/s11101-017-9534-0","article-title":"Elicitation: A stimulation of stress in vitro plant cell\/tissue cultures for enhancement of secondary metabolite production","volume":"16","author":"Narayani","year":"2017","journal-title":"Phytochem. Rev."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1007\/s11240-021-02131-1","article-title":"Biotic elicitors:a boon for the in-vitro production of plant secondary metabolites","volume":"149","author":"Bhaskar","year":"2022","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1146\/annurev.pp.46.060195.001201","article-title":"Chemoperception of microbial signals in plant cells","volume":"46","author":"Boller","year":"1995","journal-title":"Annu. Rev. Plant Physiol. Plant Mol. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Bhatia, S., Sharma, K., Dahiya, R., and Bera, T. (2015). Modern Applications of Plant Biotechnology in Pharmaceutical Sciences, Elsevier Inc.","DOI":"10.1016\/B978-0-12-802221-4.00012-1"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Malerba, M., and Cerana, R. (2016). Chitosan effects on plant systems. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17070996"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Ramirez-Estrada, K., Vidal-Limon, H., Hidalgo, D., Moyano, E., Golenioswki, M., Cusid\u00f3, R.M., and Palazon, J. (2016). Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules, 21.","DOI":"10.3390\/molecules21020182"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"100103","DOI":"10.1016\/j.stress.2022.100103","article-title":"Role of elicitors to initiate the induction of systemic resistance in plants to biotic stress","volume":"5","author":"Meena","year":"2022","journal-title":"Plant Stress"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1007\/s11240-018-1488-4","article-title":"Chitosan versus yeast extract driven elicitation for enhanced production of fragrant compound 2-hydroxy-4-methoxybenzaldehyde (2H4MB) in root tuber derived callus of Decalepis salicifolia (Bedd. Ex Hook.f.) Venter","volume":"136","author":"Ahmad","year":"2019","journal-title":"Plant Cell. Tissue Organ Cult."},{"key":"ref_29","first-page":"100054","article-title":"Efficiency of microbial bio-agents as elicitors in plant defense mechanism under biotic stress: A review","volume":"2","author":"Zehra","year":"2021","journal-title":"Curr. Res. Microb. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Iula, G., Miras-Moreno, B., Rouphael, Y., Lucini, L., and Trevisan, M. (2022). The complex metabolomics crosstalk triggered by four molecular elicitors in tomato. Plants, 11.","DOI":"10.3390\/plants11050678"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"113322","DOI":"10.1016\/j.indcrop.2021.113322","article-title":"The metabolomics reveals intraspecies variability of bioactive compounds in elicited suspension cell cultures of three Bryophyllum species","volume":"163","author":"Lucini","year":"2021","journal-title":"Ind. Crops Prod."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1002\/bit.260420416","article-title":"Plant cell suspension culture rheology","volume":"42","author":"Curtis","year":"1993","journal-title":"Biotechnol. Bioeng."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.bej.2009.02.008","article-title":"Bioreactor engineering for recombinant protein production in plant cell suspension cultures","volume":"45","author":"Huang","year":"2009","journal-title":"Biochem. Eng. J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1007\/s11240-015-0915-z","article-title":"Sucrose induced osmotic stress and photoperiod regimes enhanced the biomass and production of antioxidant secondary metabolites in shake-flask suspension cultures of Prunella vulgaris L.","volume":"124","author":"Fazal","year":"2016","journal-title":"Plant Cell. Tissue Organ Cult."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/S0141-0229(00)00292-1","article-title":"Effect of osmotic pressure on paclitaxel production in suspension cell cultures of Taxus chinensis","volume":"28","author":"Kim","year":"2001","journal-title":"Enzyme Microb. Technol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1111\/j.1365-2621.2002.tb11363.x","article-title":"Effect of sugar and nitrogen on the production of anthocyanin in cultured carrot (Daucus carota) Cells","volume":"67","author":"Narayan","year":"2002","journal-title":"J. Food Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1007\/BF03031122","article-title":"Effects of sucrose, inoculum density, auxins, and aeration volume on cell growth of Gymnema sylvestre","volume":"49","author":"Lee","year":"2006","journal-title":"J. Plant Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"24315","DOI":"10.1038\/s41598-021-03845-0","article-title":"The impact of carbon source on cell growth and the production of bioactive compounds in cell suspensions of Hancornia speciosa Gomes","volume":"11","author":"Dantas","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.procbio.2010.09.004","article-title":"Production of the anticancer drug taxol in Taxus baccata suspension cultures: A review","volume":"46","author":"Malik","year":"2011","journal-title":"Process Biochem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/0922-338X(92)90527-2","article-title":"Turbidimetric measurement of cell biomass of plant cell suspensions","volume":"73","author":"Tanaka","year":"1992","journal-title":"J. Ferment. Bioeng."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1007\/s11240-019-01593-8","article-title":"Synergistic effect of coronatine and sorbitol on artemisinin production in cell suspension culture of Artemisia annua L. Cv. Anamed","volume":"137","author":"Salehi","year":"2019","journal-title":"Plant Cell. Tissue Organ Cult."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"379","DOI":"10.3923\/biotech.2006.379.384","article-title":"Stimulatory Effects of casein hydrolysate and proline in in vitro callus induction and plant regeneration from five deep water rice (Oryza sativa L.)","volume":"5","author":"Khaleda","year":"2006","journal-title":"Biotechnology"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"15598","DOI":"10.1038\/s41598-017-15703-z","article-title":"A novel medium for enhancing callus growth of hazel (Corylus avellana L.)","volume":"7","author":"Salehi","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1007\/s11240-016-1049-7","article-title":"Elicitor induced stevioside production, in vitro shoot growth, and biomass accumulation in micropropagated Stevia rebaudiana","volume":"127","author":"Bayraktar","year":"2016","journal-title":"Plant Cell. Tissue Organ Cult."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1007\/s11240-017-1252-1","article-title":"Effect of elicitors on xanthone accumulation and biomass production in hairy root cultures of Gentiana dinarica","volume":"130","author":"Uzelac","year":"2017","journal-title":"Plant Cell. Tissue Organ Cult."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1007\/s00709-011-0264-7","article-title":"Defense\/stress responses activated by chitosan in sycamore cultured cells","volume":"249","author":"Malerba","year":"2012","journal-title":"Protoplasma"},{"key":"ref_47","first-page":"1677","article-title":"Chitosan induces plant hormones and defenses in tomato root exudates","volume":"11","author":"Arnao","year":"2020","journal-title":"Front. Plant Sci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1016\/j.plantsci.2007.01.006","article-title":"Molecular aspects of the early stages of elicitation of secondary metabolites in plants","volume":"172","author":"Vasconsuelo","year":"2007","journal-title":"Plant Sci."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1080\/10826068.2020.1830415","article-title":"Yeast extract improved biosynthesis of astragalosides in hairy root cultures of Astragalus membranaceus","volume":"51","author":"Park","year":"2021","journal-title":"Prep. Biochem. Biotechnol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"907","DOI":"10.1016\/j.apjtm.2017.08.012","article-title":"Biomass accumulation of Panax vietnamensis in cell suspension cultures varies with addition of plant growth regulators and organic additives","volume":"10","author":"Trong","year":"2017","journal-title":"Asian Pac. J. Trop. Med."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.plaphy.2016.06.034","article-title":"Induction of aromatic amino acids and phenylpropanoid compounds in Scrophularia striata Boiss. Cell Culture in Response to Chitosan-Induced Oxidative Stress","volume":"107","author":"Kamalipourazad","year":"2016","journal-title":"Plant Physiol. Biochem."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/s40529-019-0274-x","article-title":"Chitosan enhances rosmarinic acid production in shoot cultures of Melissa officinalis L. through the Induction of Methyl Jasmonate","volume":"60","author":"Shabani","year":"2019","journal-title":"Bot. Stud."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s40502-015-0139-6","article-title":"Chitosan as a promising natural compound to enhance potential physiological responses in plant: A Review","volume":"20","author":"Katiyar","year":"2015","journal-title":"Indian J. Plant Physiol."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Stasi\u0144ska-Jakubas, M., and Hawrylak-Nowak, B. (2022). Protective, biostimulating, and eliciting effects of chitosan and its derivatives on crop plants. Molecules, 27.","DOI":"10.3390\/molecules27092801"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Dalvi, S., and Suprasanna, P. (2022). Studies on radiation processed polymers mediated biochemical and molecular responses and relevance to enhancing plant productivity. Radiation-Processed Polysaccharides, Elsevier.","DOI":"10.1016\/B978-0-323-85672-0.00003-9"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1111\/j.1399-3054.1997.tb03460.x","article-title":"N-acetylchitooligosaccharides elicit expression of a single (13)-beta-glucanase gene in suspension-cultured cells from barley (Hordeum vulgare)","volume":"100","author":"Kaku","year":"1997","journal-title":"Physiol. Plant."},{"key":"ref_57","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_58","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1016\/j.sjbs.2013.12.008","article-title":"Biotic elicitor enhanced production of psoralen in suspension cultures of Psoralea corylifolia L.","volume":"21","author":"Ahmed","year":"2014","journal-title":"Saudi J. Biol. Sci."},{"key":"ref_59","first-page":"51","article-title":"Growth measurements: Estimation of cell division and cell expansion","volume":"318","year":"2006","journal-title":"Methods Mol. Biol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1006\/abio.1999.4202","article-title":"Fluorogenic substrates based on fluorinated umbelliferones for continuous assays of phosphatases and \u03b2-galactosidases","volume":"273","author":"Gee","year":"1999","journal-title":"Anal. Biochem."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"7754","DOI":"10.1073\/pnas.140132697","article-title":"Rapid and general profiling of protease specificity by using combinatorial fluorogenic substrate libraries","volume":"97","author":"Harris","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/12\/1\/190\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T17:56:38Z","timestamp":1760118998000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/12\/1\/190"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,2]]},"references-count":62,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["plants12010190"],"URL":"https:\/\/doi.org\/10.3390\/plants12010190","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,2]]}}}