{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T21:31:40Z","timestamp":1788471100472,"version":"build-2803163510"},"reference-count":62,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,2,28]],"date-time":"2024-02-28T00:00:00Z","timestamp":1709078400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Department of Education, Minority Science and Engineering Improvement Program","award":["P120A2000016"],"award-info":[{"award-number":["P120A2000016"]}]},{"name":"Department of Education, Minority Science and Engineering Improvement Program","award":["HRD-2011903"],"award-info":[{"award-number":["HRD-2011903"]}]},{"name":"Department of Education, Minority Science and Engineering Improvement Program","award":["DUE-1834046"],"award-info":[{"award-number":["DUE-1834046"]}]},{"name":"National Science Foundation Historically Black Colleges and Universities (HBCU) Undergraduate Program","award":["P120A2000016"],"award-info":[{"award-number":["P120A2000016"]}]},{"name":"National Science Foundation Historically Black Colleges and Universities (HBCU) Undergraduate Program","award":["HRD-2011903"],"award-info":[{"award-number":["HRD-2011903"]}]},{"name":"National Science Foundation Historically Black Colleges and Universities (HBCU) Undergraduate Program","award":["DUE-1834046"],"award-info":[{"award-number":["DUE-1834046"]}]},{"name":"National Science Foundation Scholarships in Science, Technology, Engineering, and Mathematics Program","award":["P120A2000016"],"award-info":[{"award-number":["P120A2000016"]}]},{"name":"National Science Foundation Scholarships in Science, Technology, Engineering, and Mathematics Program","award":["HRD-2011903"],"award-info":[{"award-number":["HRD-2011903"]}]},{"name":"National Science Foundation Scholarships in Science, Technology, Engineering, and Mathematics Program","award":["DUE-1834046"],"award-info":[{"award-number":["DUE-1834046"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Industrial hemp Cannabis sativa L. is an economically important crop mostly grown for its fiber, oil, and seeds. Due to its increasing applications in the pharmaceutical industry and a lack of knowledge of gene functions in cannabinoid biosynthesis pathways, developing an efficient transformation platform for the genetic engineering of industrial hemp has become necessary to enable functional genomic and industrial application studies. A critical step in the development of Agrobacterium tumefaciens-mediated transformation in the hemp genus is the establishment of optimal conditions for T-DNA gene delivery into different explants from which whole plantlets can be regenerated. As a first step in the development of a successful Agrobacterium tumefaciens-mediated transformation method for hemp gene editing, the factors influencing the successful T-DNA integration and expression (as measured by transient \u03b2-glucuronidase (GUS) and Green Florescent Protein (GFP) expression) were investigated. In this study, the parameters for an agroinfiltration system in hemp, which applies to the stable transformation method, were optimized. In the present study, we tested different explants, such as 1- to 3-week-old leaves, cotyledons, hypocotyls, root segments, nodal parts, and 2- to 3-week-old leaf-derived calli. We observed that the 3-week-old leaves were the best explant for transient gene expression. Fully expanded 2- to 3-week-old leaf explants, in combination with 30 min of immersion time, 60 \u00b5M silver nitrate, 0.5 \u00b5M calcium chloride, 150 \u00b5M natural phenolic compound acetosyringone, and a bacterial density of OD600nm = 0.4 resulted in the highest GUS and GFP expression. The improved method of genetic transformation established in the present study will be useful for the introduction of foreign genes of interest, using the latest technologies such as genome editing, and studying gene functions that regulate secondary metabolites in hemp.<\/jats:p>","DOI":"10.3390\/plants13050664","type":"journal-article","created":{"date-parts":[[2024,2,28]],"date-time":"2024-02-28T06:14:22Z","timestamp":1709100862000},"page":"664","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Evaluation of Parameters Affecting Agrobacterium-Mediated Transient Gene Expression in Industrial Hemp (Cannabis sativa L.)"],"prefix":"10.3390","volume":"13","author":[{"given":"Tasnim","family":"Mohammad","sequence":"first","affiliation":[{"name":"Center for Biotechnology, Department of Agricultural Sciences, Fort Valley State University, 113, Alva Tabor Building, Fort Valley, GA 31030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rishikesh","family":"Ghogare","sequence":"additional","affiliation":[{"name":"Department of Horticultural Sciences, Texas A&M University, College Station, TX 77843, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lauren B.","family":"Morton","sequence":"additional","affiliation":[{"name":"Center for Biotechnology, Department of Agricultural Sciences, Fort Valley State University, 113, Alva Tabor Building, Fort Valley, GA 31030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amit","family":"Dhingra","sequence":"additional","affiliation":[{"name":"Department of Horticultural Sciences, Texas A&M University, College Station, TX 77843, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shobha","family":"Potlakayala","sequence":"additional","affiliation":[{"name":"The Central Pennsylvania Research and Teaching Laboratory for Biofuels, Penn State Harrisburg, Middletown, PA 17057, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sairam","family":"Rudrabhatla","sequence":"additional","affiliation":[{"name":"The Central Pennsylvania Research and Teaching Laboratory for Biofuels, Penn State Harrisburg, Middletown, PA 17057, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarwan K.","family":"Dhir","sequence":"additional","affiliation":[{"name":"Center for Biotechnology, Department of Agricultural Sciences, Fort Valley State University, 113, Alva Tabor Building, Fort Valley, GA 31030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"958","DOI":"10.3389\/fpls.2020.00958","article-title":"Propagation of Cannabis for clinical research: An approach towards a modern herbal medicinal products development","volume":"11","author":"Chandra","year":"2020","journal-title":"Front. Plant Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3504","DOI":"10.1038\/s41598-020-60323-9","article-title":"Establishment and optimization of hemp (Cannabis sativa L.) agroinfiltration system for gene expression and silencing studies","volume":"10","author":"Deguchi","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/j.tips.2009.07.006","article-title":"Non-psychotropic plant cannabinoids: New therapeutic opportunities from an ancient herb","volume":"30","author":"Izzo","year":"2009","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Gon\u00e7alves, J., Rosado, T., Soares, S., Sim\u00e3o, A.Y., Caramelo, D., Lu\u00eds, \u00c2., Fern\u00e1ndez, N., Barroso, M., Gallardo, E., and Duarte, A.P. (2019). Cannabis and its secondary metabolites: Their use as therapeutic drugs, toxicological aspects, and analytical determination. Medicines, 6.","DOI":"10.3390\/medicines6010031"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Govindarajan, R.K., Mishra, A.K., Cho, K.H., Kim, K.H., Yoon, K.M., and Baek, K.H. (2023). Biosynthesis of Phytocannabinoids and Structural Insights: A Review. Metabolites, 13.","DOI":"10.3390\/metabo13030442"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1154332","DOI":"10.3389\/fpls.2023.1154332","article-title":"Challenges, and potentials of new breeding techniques in Cannabis sativa L","volume":"8","author":"Ingvardsen","year":"2023","journal-title":"Front. Plant Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"580621","DOI":"10.3389\/fpls.2020.580621","article-title":"Metabolic Engineering Strategies of Industrial Hemp (Cannabis sativa L.): A Brief Review of the Advances and Challenges","volume":"8","author":"Deguchi","year":"2020","journal-title":"Front. Plant Sci."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Hesami, M., Baiton, A., Alizadeh, M., Pepe, M., Torkamaneh, D., and Jones, A.M.P. (2021). Advances and perspectives in tissue culture and genetic engineering of Cannabis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22115671"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Sun, X. (2023). Research progress on Cannabinoids in Cannabis (Cannabis sativa L.) in China. Molecules, 29.","DOI":"10.3390\/molecules28093806"},{"key":"ref_10","unstructured":"Gilchrist, E.J., Wang, S., and Quilichini, T.D. (2023). Genomics and the Global Bioeconomy, Academic Press."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.1038\/s41477-022-01295-8","article-title":"Recent advances in crop transformation technologies","volume":"8","author":"Chen","year":"2022","journal-title":"Nat. Plants"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Su, W., Xu, M., Radani, Y., and Yang, L. (2023). Technological Development and Application of Plant Genetic Transformation. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241310646"},{"key":"ref_13","first-page":"15","article-title":"Agrobacterium-mediated plant transformation: Biology and applications","volume":"20","author":"Hwang","year":"2017","journal-title":"Arab. Book"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1007\/s11248-021-00239-w","article-title":"Genome editing reagent delivery in plants","volume":"30","author":"Ghogare","year":"2021","journal-title":"Transgenic Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"100028","DOI":"10.1016\/j.xplc.2020.100028","article-title":"A Highly Efficient Agrobacterium-Mediated Method for Transient Gene Expression and Functional Studies in Multiple Plant Species","volume":"1","author":"Zhang","year":"2020","journal-title":"Plant Commun."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1139\/cjb-2023-0084","article-title":"Agrobacterium-mediated transformation of THC-containing Cannabis sativa L. yields a high frequency of transgenic calli expressing bialaphos resistance and non-expressor of PR1 (NPR1) genes","volume":"101","author":"Holmes","year":"2023","journal-title":"Botany"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"627240","DOI":"10.3389\/fpls.2021.627240","article-title":"Medical Cannabis and industrial hemp tissue culture: Present status and future potential","volume":"3","author":"Adhikary","year":"2021","journal-title":"Front. Plant Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"113360","DOI":"10.1016\/j.indcrop.2021.113360","article-title":"Protoplast isolation, transient transformation, and flow-cytometric analysis of reporter-gene activation in Cannabis sativa L","volume":"164","author":"Beard","year":"2021","journal-title":"Ind. Crops Prod."},{"key":"ref_19","first-page":"1040","article-title":"Development of a robust transient expression screening system in protoplasts of Cannabis","volume":"57","author":"Mohamaden","year":"2021","journal-title":"Vitr. Cell. Dev. Biol. Plant"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1007\/s11816-022-00786-6","article-title":"Establishment of efficient Cannabis (Cannabis sativa L.) protoplast isolation and transient expression condition","volume":"16","author":"Kim","year":"2022","journal-title":"Plant Biotechnol. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1139\/cjb-2021-0178","article-title":"A versatile protoplast system and its application in Cannabis sativa L","volume":"101","author":"Zhu","year":"2023","journal-title":"Botany"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1079\/IVP2003454","article-title":"Tissue culture and Agrobacterium-mediated transformation of Hemp (Cannabis sativa L.)","volume":"39","author":"Feeney","year":"2003","journal-title":"Vitro Cell. Dev. Biol. Plant."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1080\/17429145.2012.746399","article-title":"Agrobacterium infection of Hemp (Cannabis sativa L.): Establishment of hairy root cultures","volume":"8","author":"Wahby","year":"2013","journal-title":"J. Plant Interact."},{"key":"ref_24","unstructured":"Roscow, R.F. (2017). Enhanced Cannabis Plants and Methods of Making and Using the Same. (2017\/1810180 A1), Patent with International Publication Number WO."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"921970","DOI":"10.3389\/fpls.2022.921970","article-title":"In planta, female flower agroinfiltration alters the Cannabinoid composition in industrial hemp (Cannabis sativa L.)","volume":"13","author":"Deguchi","year":"2022","journal-title":"Front. Plant Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.jbiotec.2020.12.014","article-title":"Nanoparticle-based genetic transformation of Cannabis sativa L","volume":"20","author":"Ahmed","year":"2021","journal-title":"J. Biotechnol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"113691","DOI":"10.1016\/j.indcrop.2021.113691","article-title":"A novel and rapid method for Agrobacterium-mediated production of stably transformed Cannabis sativa L. Plants","volume":"170","author":"Gramazio","year":"2021","journal-title":"Ind. Crops Prod."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1979","DOI":"10.1111\/pbi.13611","article-title":"Establishment of an Agrobacterium-mediated genetic transformation and CRISPR\/Cas9-mediated targeted mutagenesis in hemp (Cannabis sativa L.)","volume":"19","author":"Zhang","year":"2021","journal-title":"Plant Biotechnol. J."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Monthony, A.S., Page, S.R., Hesami, M., and Jones, A.M.P. (2021). The past, present, and future of Cannabis sativa L. tissue culture. Plants, 19.","DOI":"10.20944\/preprints202012.0766.v1"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"684192","DOI":"10.3389\/fpls.2021.684192","article-title":"Agrobacterium VirE2 protein modulates plant gene expression and mediates transformation from its location outside the nucleus","volume":"12","author":"Lapham","year":"2021","journal-title":"Front. Plant Sci."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Sivanandhan, G., Kapil, D.G., Theboral, J., Selvaraj, N., Ganapathi, A., and Manickavasagam, M. (2015). Sonication, vacuum infiltration, and thiol compounds enhance the Agrobacterium-mediated transformation frequency of Withania somnifera (L.) Dunal. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0124693"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/s13007-018-0343-2","article-title":"Improving agroinfiltration-based transient gene expression in Nicotiana benthamiana","volume":"14","author":"Norkunas","year":"2018","journal-title":"Plant Methods"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Xi, J., Patel, M., Dong, S., Que, Q., and Qu, R. (2018). Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus. BMC Biotechnol., 18.","DOI":"10.1186\/s12896-018-0459-5"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"61","DOI":"10.5010\/JPB.2022.49.1.061","article-title":"Optimization of different factors for an Agrobacterium-mediated genetic transformation system using embryo axis explants of chickpea (Cicer arietinum L.)","volume":"49","author":"Sadhu","year":"2022","journal-title":"J. Plant Biotechnol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1111\/tpj.15202","article-title":"Developing a rapid and highly efficient cowpea regeneration, transformation, and genome editing system using embryonic axis explants","volume":"106","author":"Che","year":"2021","journal-title":"Plant J."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Liu, H., Zhao, J., Chen, F., Wu, Z., Tan, J., Nguyen, N.H., Cheng, Z., and Weng, Y. (2023). Improving Agrobacterium tumefaciens-mediated genetic transformation for gene function studies and mutagenesis in cucumber (Cucumis sativus L.). Genes, 14.","DOI":"10.3390\/genes14030601"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"831","DOI":"10.1111\/j.1365-2958.1997.mmi501.x","article-title":"Control of virulence gene expression by plant calcium in the phytopathogen Erwinia carotovora","volume":"25","author":"Flego","year":"1997","journal-title":"Mol. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/s10725-004-6126-y","article-title":"Silver nitrate and aminoethoxyvinylglycine affect Agrobacterium-mediated apple transformation","volume":"45","author":"Seong","year":"2005","journal-title":"Plant Growth Regul."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"468","DOI":"10.3923\/ajps.2006.468.480","article-title":"Transient expression of gusA and gfp gene in Agrobacterium-mediated banana transformation using single tiny meristematic bud","volume":"5","author":"Subramaniam","year":"2006","journal-title":"Asian J. Plant Sci."},{"key":"ref_40","first-page":"3424","article-title":"Optimization of transient green fluorescent protein (GFP) gene expression in Phalaenopsis violacea orchid mediated by Agrobacterium tumefaciens-mediated transformation system","volume":"4","author":"Julkifle","year":"2010","journal-title":"Aust. J. Basic Appl. Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1186\/s13007-018-0360-1","article-title":"Agrobacterium-mediated transformation systems of Primula vulgaris","volume":"26","author":"Hayta","year":"2018","journal-title":"Plant Methods"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1780037","DOI":"10.1080\/15592324.2020.1780037","article-title":"Transient expression of the \u03b2-glucuronidase gene in Cannabis sativa L. varieties","volume":"15","author":"Sorokin","year":"2020","journal-title":"Plant Signal. Behav."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Sorokin, A., Yadav, N.S., Gaudet, D., and Kovalchuk, I. (2021). Development and standardization of rapid and efficient seed germination protocol for Cannabis sativa L. Bio Protoc., 11.","DOI":"10.21769\/BioProtoc.3875"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"971","DOI":"10.1104\/pp.115.3.971","article-title":"Genetic transformation of wheat mediated by Agrobacterium tumefaciens","volume":"115","author":"Cheng","year":"1997","journal-title":"Plant Physiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1023\/A:1005847615493","article-title":"Transformation of rice mediated by Agrobacterium tumefaciens","volume":"35","author":"Hiei","year":"1997","journal-title":"Plant Mol. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2281","DOI":"10.1007\/s00299-011-1133-8","article-title":"Improved Agrobacterium-mediated transformation of cowpea via sonication and vacuum infiltration","volume":"30","author":"Bakshi","year":"2011","journal-title":"Plant Cell Rep."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1016\/j.biotechadv.2015.03.012","article-title":"Transient plant transformation mediated by Agrobacterium tumefaciens: Principles, methods, and applications","volume":"1","author":"Krenek","year":"2015","journal-title":"Biotechnol Adv."},{"key":"ref_48","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 bioassays with tobacco tissue cultures","volume":"15","author":"Murashige","year":"1962","journal-title":"Physiol. Plant."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/0014-4827(68)90403-5","article-title":"Nutrient requirements of suspension cultures of soybean root cells","volume":"50","author":"Gamborg","year":"1968","journal-title":"Exp. Cell Res."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Sangra, A., Shahin, L., and Dhir, S.K. (2019). Long-term maintainable somatic embryogenesis system in Alfalfa (Medicago sativa) using leaf explants: Embryogenic sustainability approach. Plants, 9.","DOI":"10.3390\/plants8080278"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1007\/s00299-009-0717-z","article-title":"Optimization of conditions for transient Agrobacterium-mediated gene expression assays in Arabidopsis","volume":"28","author":"Kim","year":"2009","journal-title":"Plant Cell Rep."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1497","DOI":"10.1007\/s00709-020-01524-x","article-title":"An attempt to establish an Agrobacterium-mediated transient expression system in medicinal plants","volume":"257","author":"Xia","year":"2020","journal-title":"Protoplasma"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"584011","DOI":"10.3389\/fpls.2020.584011","article-title":"A rapid and highly efficient method for transient gene expression in rice plants","volume":"11","author":"Burman","year":"2020","journal-title":"Front. Plant Sci."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1007\/s00299-014-1684-6","article-title":"The Brachypodium distachyon BdWRKY36 gene confers tolerance to drought stress in transgenic tobacco plants","volume":"34","author":"Sun","year":"2015","journal-title":"Plant Cell Rep."},{"key":"ref_55","first-page":"459","article-title":"Agroinfiltration for transient gene expression in floral tissues of Dendrobium Sonia \u2018Earsakul\u2019","volume":"10","author":"Pinthong","year":"2014","journal-title":"J. Agric. Technol."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Shafaghi, M., Maktoobian, S., Rasouli, R., Howaizi, N., Ofoghi, H., and Ehsani, P. (2018). Transient expression of biologically active anti-rabies virus monoclonal antibody in tobacco leaves. Iran J. Biotechnol., 16.","DOI":"10.21859\/ijb.1774"},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Dan, Y., Baxter, A., Zhang, S., Pantazis, C.J., and Veilleux, R.E. (2010). Development of efficient plant regeneration and transformation system for impatiens using Agrobacterium tumefaciens and multiple bud cultures as explants. BMC Plant Biol., 9.","DOI":"10.1186\/1471-2229-10-165"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1128\/jb.62.3.293-300.1951","article-title":"Studies on lysogenesis","volume":"62","author":"Bertani","year":"1951","journal-title":"J. Bacteriol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1038\/342837a0","article-title":"The GUS reporter gene system","volume":"342","author":"Jefferson","year":"1989","journal-title":"Nature"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/0168-9452(90)90042-M","article-title":"An improved assay for \u03b2-glucuronidase in transformed cells: Methanol almost completely suppresses a putative endogenous \u03b2-glucuronidase activity","volume":"70","author":"Kosugi","year":"1990","journal-title":"Plant Sci."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1007\/978-1-0716-3299-4_18","article-title":"The GUS reporter system in flower development studies","volume":"2686","author":"Mudunkothge","year":"2023","journal-title":"Methods Mol. Biol."},{"key":"ref_62","unstructured":"Abdi, H., and Williams, L.J. (2010). Encyclopedia of Research Design, Sage."}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/13\/5\/664\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:06:11Z","timestamp":1760105171000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/13\/5\/664"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,28]]},"references-count":62,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,3]]}},"alternative-id":["plants13050664"],"URL":"https:\/\/doi.org\/10.3390\/plants13050664","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,28]]}}}