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Development of a Direct in vitro Plant Regeneration Protocol From Cannabis sativa L. Seedling Explants: Developmental Morphology of Shoot Regeneration and Ploidy Level of Regenerated Plants

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Development of a Direct in vitro Plant Regeneration Protocol From Cannabis sativa L. Seedling Explants: Developmental Morphology of Shoot Regeneration and Ploidy Level of Regenerated Plants

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dc.contributor.author Galán-Ávila, Alberto es_ES
dc.contributor.author García-Fortea, Edgar es_ES
dc.contributor.author Prohens Tomás, Jaime es_ES
dc.contributor.author Herraiz García, Francisco Javier es_ES
dc.date.accessioned 2021-06-16T03:30:40Z
dc.date.available 2021-06-16T03:30:40Z
dc.date.issued 2020-05-27 es_ES
dc.identifier.uri http://hdl.handle.net/10251/168035
dc.description.abstract [EN] In vitro shoot regeneration can efficiently contribute to the improvement of recalcitrant Cannabis sativa L. We aimed at developing a highly efficient protocol for in vitro direct regeneration of C. sativa plants from different explants (cotyledon, hypocotyl, and true leaf) from seedlings of monoecious C. sativa short-day varieties Ferimon, Felina32, Fedora17, and USO31, together with dioecious neutral-day variety Finola. Ten regeneration media, including already published protocols, and self-designed combinations of plant growth regulators were tested. The developmental morphology since germination of seeds to the development of rooted plantlets was followed. Additionally, the ploidy level of explants and in vitro regenerants was analyzed. We concluded that hypocotyl is the best explant for in vitro direct regeneration of C. sativa plants with 49.45% of responding explants, while cotyledon and true leaf had a poor response with, respectively, 4.70 and 0.42% of explants developing plantlets. In terms of shoot regeneration, we found significant differences among the culture media evaluated and the varieties studied. Overall, the best regeneration media were ZEARIB 2.0 (mg/L) and ZEARIB 1.0 (mg/L) C NAA 0.02 (mg/L) with 66.67% of responding hypocotyls. Amazingly, hypocotyls cultured in medium without plant growth regulators showed an excellent response (61.54% of responding hypocotyls) and spontaneous rooting of regenerants (17.94%). In vitro regenerated plants were acclimatized just 6 weeks after culture initiation. The developmental morphology study suggests that regenerated shoots originate from pericycle cells adjacent to xylem poles. Polysomaty was detected in hypocotyls and cotyledons of all varieties studied, and diploid (>80%) and mixoploid (with diploid and tetraploid cells) plants were regenerated. Our protocol allows a high shoot organogenesis efficiency in different C. sativa varieties. The fact that a significant percentage of plants are mixoploid may provide an alternative way to develop polyploids in C. sativa. Our results show that direct in vitro regeneration may make a significant contribution to the development of improved C. sativa materials for medical applications. es_ES
dc.language Inglés es_ES
dc.publisher Frontiers Media SA es_ES
dc.relation.ispartof Frontiers in Plant Science es_ES
dc.rights Reconocimiento (by) es_ES
dc.subject Cannabinoids es_ES
dc.subject Hemp es_ES
dc.subject Hypocotyl es_ES
dc.subject Micropropagation es_ES
dc.subject Polyploidization es_ES
dc.subject Polysomaty es_ES
dc.subject Shoot organogenesis es_ES
dc.subject.classification GENETICA es_ES
dc.title Development of a Direct in vitro Plant Regeneration Protocol From Cannabis sativa L. Seedling Explants: Developmental Morphology of Shoot Regeneration and Ploidy Level of Regenerated Plants es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.3389/fpls.2020.00645 es_ES
dc.rights.accessRights Abierto es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Universitario de Conservación y Mejora de la Agrodiversidad Valenciana - Institut Universitari de Conservació i Millora de l'Agrodiversitat Valenciana es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia es_ES
dc.description.bibliographicCitation Galán-Ávila, A.; García-Fortea, E.; Prohens Tomás, J.; Herraiz García, FJ. (2020). Development of a Direct in vitro Plant Regeneration Protocol From Cannabis sativa L. Seedling Explants: Developmental Morphology of Shoot Regeneration and Ploidy Level of Regenerated Plants. Frontiers in Plant Science. 11:1-15. https://doi.org/10.3389/fpls.2020.00645 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion https://doi.org/10.3389/fpls.2020.00645 es_ES
dc.description.upvformatpinicio 1 es_ES
dc.description.upvformatpfin 15 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 11 es_ES
dc.identifier.eissn 1664-462X es_ES
dc.identifier.pmid 32670304 es_ES
dc.identifier.pmcid PMC7326123 es_ES
dc.relation.pasarela S\413989 es_ES
dc.description.references Adelberg, J. W., Rhodes, B. B., & Skorupska, H. T. (1993). GENERATING TETRAPLOID MELONS IN TISSUE CULTURE. Acta Horticulturae, (336), 373-380. doi:10.17660/actahortic.1993.336.49 es_ES
dc.description.references Andre, C. M., Hausman, J.-F., & Guerriero, G. (2016). Cannabis sativa: The Plant of the Thousand and One Molecules. Frontiers in Plant Science, 7. doi:10.3389/fpls.2016.00019 es_ES
dc.description.references Atta, R., Laurens, L., Boucheron-Dubuisson, E., Guivarc’h, A., Carnero, E., Giraudat-Pautot, V., … Chriqui, D. (2009). Pluripotency of Arabidopsis xylem pericycle underlies shoot regeneration from root and hypocotyl explants grownin vitro. The Plant Journal, 57(4), 626-644. doi:10.1111/j.1365-313x.2008.03715.x es_ES
dc.description.references Beck, E. H. (1996). Regulation of shoot/root ratio by cytokinins from roots inUrtica dioica: Opinion. Plant and Soil, 185(1), 1-12. doi:10.1007/bf02257560 es_ES
dc.description.references Beeckman, T., & De Smet, I. (2014). Pericycle. Current Biology, 24(10), R378-R379. doi:10.1016/j.cub.2014.03.031 es_ES
dc.description.references Behr, M., Legay, S., Žižková, E., Motyka, V., Dobrev, P. I., Hausman, J.-F., … Guerriero, G. (2016). Studying Secondary Growth and Bast Fiber Development: The Hemp Hypocotyl Peeks behind the Wall. Frontiers in Plant Science, 7. doi:10.3389/fpls.2016.01733 es_ES
dc.description.references Behr, M., Sergeant, K., Leclercq, C. C., Planchon, S., Guignard, C., Lenouvel, A., … Guerriero, G. (2018). Insights into the molecular regulation of monolignol-derived product biosynthesis in the growing hemp hypocotyl. BMC Plant Biology, 18(1). doi:10.1186/s12870-017-1213-1 es_ES
dc.description.references Breslavetz, L. (1932). Polyploide Mitosen bei Cannabis sativa L. II. Planta, 17(3), 644-649. doi:10.1007/bf01909774 es_ES
dc.description.references Bubner, B., Gase, K., Berger, B., Link, D., & Baldwin, I. T. (2006). Occurrence of tetraploidy in Nicotiana attenuata plants after Agrobacterium-mediated transformation is genotype specific but independent of polysomaty of explant tissue. Plant Cell Reports, 25(7), 668-675. doi:10.1007/s00299-005-0111-4 es_ES
dc.description.references Cascio, M. G., Pertwee, R. G., & Marini, P. (2017). The Pharmacology and Therapeutic Potential of Plant Cannabinoids. Cannabis sativa L. - Botany and Biotechnology, 207-225. doi:10.1007/978-3-319-54564-6_9 es_ES
dc.description.references Chaohua, C., Gonggu, Z., Lining, Z., Chunsheng, G., Qing, T., Jianhua, C., … Jianguang, S. (2016). A rapid shoot regeneration protocol from the cotyledons of hemp (Cannabis sativa L.). Industrial Crops and Products, 83, 61-65. doi:10.1016/j.indcrop.2015.12.035 es_ES
dc.description.references Colijn-Hooymans, C. M., Hakkert, J. C., Jansen, J., & Custers, J. B. M. (1994). Competence for regeneration of cucumber cotyledons is restricted to specific developmental stages. Plant Cell, Tissue and Organ Culture, 39(3), 211-217. doi:10.1007/bf00035972 es_ES
dc.description.references D’Amato, F. (1952). Polyploidy in the Differentiation and Function of Tissues and Cells in Plants. Caryologia, 4(3), 311-358. doi:10.1080/00087114.1952.10797544 es_ES
dc.description.references D’Amato, F. (1964). Endopolyploidy as a Factor in Plant Tissue Development. Caryologia, 17(1), 41-52. doi:10.1080/00087114.1964.10796115 es_ES
dc.description.references DETREZ, C., TETU, T., SANGWAN, R. S., & SANGWAN-NORREEL, B. S. (1988). Direct Organogenesis from Petiole and Thin Cell Layer Explants in Sugar Beet CulturedIn Vitro. Journal of Experimental Botany, 39(7), 917-926. doi:10.1093/jxb/39.7.917 es_ES
dc.description.references Dpooležel, J., Binarová, P., & Lcretti, S. (1989). Analysis of Nuclear DNA content in plant cells by Flow cytometry. Biologia Plantarum, 31(2), 113-120. doi:10.1007/bf02907241 es_ES
dc.description.references Ervin, C. D. (1939). Polysomaty in Cucumis Melo. Proceedings of the National Academy of Sciences, 25(7), 335-338. doi:10.1073/pnas.25.7.335 es_ES
dc.description.references Ervin, C. D. (1941). A STUDY OF POLYSOMATY IN CUCUMIS MELO. American Journal of Botany, 28(2), 113-124. doi:10.1002/j.1537-2197.1941.tb07950.x es_ES
dc.description.references Evans, D. A., & Bravo, J. E. (1986). Phenotypic and Genotypic Stability of Tissue Cultured Plants. Current Plant Science and Biotechnology in Agriculture, 73-94. doi:10.1007/978-94-009-4444-2_6 es_ES
dc.description.references Ezura, H., Nishimiya, S., & Kasumi, M. (1993). Efficient regeneration of plants independent of exogeneous growth regulators in bell pepper (Capsicum annumm L.). Plant Cell Reports, 12(12). doi:10.1007/bf00233418 es_ES
dc.description.references Feeney, M., & Punja, Z. K. (2003). Tissue culture and Agrobacterium-mediated transformation of hemp (Cannabis sativa L.). In Vitro Cellular & Developmental Biology - Plant, 39(6), 578-585. doi:10.1079/ivp2003454 es_ES
dc.description.references Feeney, M., & Punja, Z. K. (2014). Hemp (Cannabis sativa L.). Agrobacterium Protocols, 319-329. doi:10.1007/978-1-4939-1658-0_25 es_ES
dc.description.references Feeney, M., & Punja, Z. K. (2017). The Role of Agrobacterium-Mediated and Other Gene-Transfer Technologies in Cannabis Research and Product Development. Cannabis sativa L. - Botany and Biotechnology, 343-363. doi:10.1007/978-3-319-54564-6_16 es_ES
dc.description.references Cardoso-Furlan, F., Gavilan, N. H., Zichner-Zorz, A., Oliveira, L. S. de, Konzen, E. R., & Ebling-Brondani, G. (2018). Active chlorine and charcoal affect the in vitro culture of Bambusa vulgaris. Bosque (Valdivia), 39(1), 61-70. doi:10.4067/s0717-92002018000100061 es_ES
dc.description.references García-Fortea, E., Lluch-Ruiz, A., Pineda-Chaza, B. J., García-Pérez, A., Bracho-Gil, J. P., Plazas, M., … Prohens, J. (2020). A highly efficient organogenesis protocol based on zeatin riboside for in vitro regeneration of eggplant. BMC Plant Biology, 20(1). doi:10.1186/s12870-019-2215-y es_ES
dc.description.references Iannicelli, J., Guariniello, J., Tossi, V. E., Regalado, J. J., Di Ciaccio, L., van Baren, C. M., … Escandón, A. S. (2020). The «polyploid effect» in the breeding of aromatic and medicinal species. Scientia Horticulturae, 260, 108854. doi:10.1016/j.scienta.2019.108854 es_ES
dc.description.references Ihaka, R., & Gentleman, R. (1996). R: A Language for Data Analysis and Graphics. Journal of Computational and Graphical Statistics, 5(3), 299-314. doi:10.1080/10618600.1996.10474713 es_ES
dc.description.references LaRue, C. D. (1933). Regeneration in Mutilated Seedlings. Proceedings of the National Academy of Sciences, 19(1), 53-63. doi:10.1073/pnas.19.1.53 es_ES
dc.description.references Lata, H., Chandra, S., Khan, I., & ElSohly, M. A. (2008). Thidiazuron-induced high-frequency direct shoot organogenesis of Cannabis sativa L. In Vitro Cellular & Developmental Biology - Plant, 45(1), 12-19. doi:10.1007/s11627-008-9167-5 es_ES
dc.description.references Lata, H., Chandra, S., Khan, I., & ElSohly, M. (2010). High Frequency Plant Regeneration from Leaf Derived Callus of HighΔ9-Tetrahydrocannabinol YieldingCannabis sativaL. Planta Medica, 76(14), 1629-1633. doi:10.1055/s-0030-1249773 es_ES
dc.description.references Lata, H., Chandra, S., Khan, I. A., & ElSohly, M. A. (2016). In Vitro Propagation of Cannabis sativa L. and Evaluation of Regenerated Plants for Genetic Fidelity and Cannabinoids Content for Quality Assurance. Protocols for In Vitro Cultures and Secondary Metabolite Analysis of Aromatic and Medicinal Plants, Second Edition, 275-288. doi:10.1007/978-1-4939-3332-7_19 es_ES
dc.description.references Lata, H., Chandra, S., Techen, N., Khan, I. A., & ElSohly, M. A. (2016). In vitro mass propagation of Cannabis sativa L.: A protocol refinement using novel aromatic cytokinin meta-topolin and the assessment of eco-physiological, biochemical and genetic fidelity of micropropagated plants. Journal of Applied Research on Medicinal and Aromatic Plants, 3(1), 18-26. doi:10.1016/j.jarmap.2015.12.001 es_ES
dc.description.references Lata, H., Chandra, S., Khan, I. A., & ElSohly, M. A. (2017). Micropropagation of Cannabis sativa L.—An Update. Cannabis sativa L. - Botany and Biotechnology, 285-297. doi:10.1007/978-3-319-54564-6_13 es_ES
dc.description.references Ranalli, P. (1999). Advances in Hemp Research. doi:10.1201/9781498705820 es_ES
dc.description.references Mansouri, H., & Bagheri, M. (2017). Induction of Polyploidy and Its Effect on Cannabis sativa L. Cannabis sativa L. - Botany and Biotechnology, 365-383. doi:10.1007/978-3-319-54564-6_17 es_ES
dc.description.references Mhatre, M., Bapat, V. A., & Rao, P. S. (1985). Regeneration of plants from the culture of leaves and axillary buds in mulberry (Morus indica L.). Plant Cell Reports, 4(2), 78-80. doi:10.1007/bf00269211 es_ES
dc.description.references Miller, R. H. (1959). MORPHOLOGY OF HUMULUS LUPULUS. II. SECONDARY GROWTH IN THE ROOT AND SEEDLING VASCULARIZATION. American Journal of Botany, 46(4), 269-277. doi:10.1002/j.1537-2197.1959.tb07012.x es_ES
dc.description.references Minocha, S. C. (1987). Plant Growth Regulators and Morphogenesis in Cell and Tissue Culture of Forest Trees. Forestry Sciences, 50-66. doi:10.1007/978-94-017-0994-1_4 es_ES
dc.description.references Mishchenko, S., Mokher, J., Laiko, I., Burbulis, N., Kyrychenko, H., & Dudukova, S. (2017). Phenological growth stages of hemp (Cannabis sativa L.): codification and description according to the BBCH scale. Žemės ūkio mokslai, 24(2). doi:10.6001/zemesukiomokslai.v24i2.3496 es_ES
dc.description.references Murashige, T., & Skoog, F. (1962). A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiologia Plantarum, 15(3), 473-497. doi:10.1111/j.1399-3054.1962.tb08052.x es_ES
dc.description.references Parsons, J. L., Martin, S. L., James, T., Golenia, G., Boudko, E. A., & Hepworth, S. R. (2019). Polyploidization for the Genetic Improvement of Cannabis sativa. Frontiers in Plant Science, 10. doi:10.3389/fpls.2019.00476 es_ES
dc.description.references Ramírez-Mosqueda, M. A., & Iglesias-Andreu, L. G. (2015). Indirect organogenesis and assessment of somaclonal variation in plantlets of Vanilla planifolia Jacks. Plant Cell, Tissue and Organ Culture (PCTOC), 123(3), 657-664. doi:10.1007/s11240-015-0868-2 es_ES
dc.description.references RAMSAY, G., & KUMAR, A. (1990). Transformation ofVicia fabaCotyledon and Stem Tissues7Agrobacterium rhizogenes: Infectivity and Cytological Studies. Journal of Experimental Botany, 41(7), 841-847. doi:10.1093/jxb/41.7.841 es_ES
dc.description.references Recupero, G. R., Russo, G., & Recupero, S. (2005). New Promising Citrus Triploid Hybrids Selected from Crosses between Monoembryonic Diploid Female and Tetraploid Male Parents. HortScience, 40(3), 516-520. doi:10.21273/hortsci.40.3.516 es_ES
dc.description.references Ren, Y., Bang, H., Gould, J., Rathore, K. S., Patil, B. S., & Crosby, K. M. (2012). Shoot regeneration and ploidy variation in tissue culture of honeydew melon (Cucumis melo L. inodorus). In Vitro Cellular & Developmental Biology - Plant, 49(2), 223-229. doi:10.1007/s11627-012-9482-8 es_ES
dc.description.references RICHEZ-DUMANOIS, C., BRAUT-BOUCHER, F., COSSON, L., & PARIS, M. (1986). Multiplication végétativein vitrodu chanvre (Cannabis sativa L.). Application à la conserva- tion des clones sélectionnés. Agronomie, 6(5), 487-495. doi:10.1051/agro:19860510 es_ES
dc.description.references Sairam Reddy, P., Rodrigues, R., & Rajasekharan, R. (2001). Plant Cell, Tissue and Organ Culture, 66(3), 183-188. doi:10.1023/a:1010697813852 es_ES
dc.description.references Silvarolla, M. B., Mazzafera, P., Lima, M. M. A. de, Medina Filho, H. P., & Fazuoli, L. C. (1999). Ploidy level and caffeine content in leaves of Coffea. Scientia Agricola, 56(3), 661-663. doi:10.1590/s0103-90161999000300021 es_ES
dc.description.references Sliwinska, E., & Lukaszewska, E. (2005). Polysomaty in growing in vitro sugar-beet (Beta vulgaris L.) seedlings of different ploidy level. Plant Science, 168(4), 1067-1074. doi:10.1016/j.plantsci.2004.12.003 es_ES
dc.description.references Smýkalová, I., Vrbová, M., Cvečková, M., Plačková, L., Žukauskaitė, A., Zatloukal, M., … Griga, M. (2019). The effects of novel synthetic cytokinin derivatives and endogenous cytokinins on the in vitro growth responses of hemp (Cannabis sativa L.) explants. Plant Cell, Tissue and Organ Culture (PCTOC), 139(2), 381-394. doi:10.1007/s11240-019-01693-5 es_ES
dc.description.references Su, Y.-H., Liu, Y.-B., & Zhang, X.-S. (2011). Auxin–Cytokinin Interaction Regulates Meristem Development. Molecular Plant, 4(4), 616-625. doi:10.1093/mp/ssr007 es_ES
dc.description.references Tanimoto, S., & Harada, H. (1984). Roles of Auxin and Cytokinin in Organogenesis in Torenia Stem Segments Cultured in vitro. Journal of Plant Physiology, 115(1), 11-18. doi:10.1016/s0176-1617(84)80046-2 es_ES
dc.description.references Urits, I., Borchart, M., Hasegawa, M., Kochanski, J., Orhurhu, V., & Viswanath, O. (2019). An Update of Current Cannabis-Based Pharmaceuticals in Pain Medicine. Pain and Therapy, 8(1), 41-51. doi:10.1007/s40122-019-0114-4 es_ES
dc.description.references Van den Bulk, R. W., Löffler, H. J. M., Lindhout, W. H., & Koornneef, M. (1990). Somaclonal variation in tomato: effect of explant source and a comparison with chemical mutagenesis. Theoretical and Applied Genetics, 80(6), 817-825. doi:10.1007/bf00224199 es_ES
dc.description.references Van Hieu, P. (2019). Polyploid Gene Expression and Regulation in Polysomic Polyploids. American Journal of Plant Sciences, 10(08), 1409-1443. doi:10.4236/ajps.2019.108101 es_ES
dc.description.references Vieira, L. M., Rocha, D. I., Taquetti, M. F., da Silva, L. C., de Campos, J. M. S., Viccini, L. F., & Otoni, W. C. (2014). In vitro plant regeneration of Passiflora setacea D.C. (Passifloraceae): the influence of explant type, growth regulators, and incubation conditions. In Vitro Cellular & Developmental Biology - Plant, 50(6), 738-745. doi:10.1007/s11627-014-9650-0 es_ES
dc.description.references Wahby, I., Caba, J. M., & Ligero, F. (2013). Agrobacteriuminfection of hemp (Cannabis sativaL.): establishment of hairy root cultures. Journal of Plant Interactions, 8(4), 312-320. doi:10.1080/17429145.2012.746399 es_ES
dc.description.references Wahby, I., Caba, J. M., & Ligero, F. (2017). Hairy Root Culture as a Biotechnological Tool in C. sativa. Cannabis sativa L. - Botany and Biotechnology, 299-317. doi:10.1007/978-3-319-54564-6_14 es_ES
dc.description.references Wielgus, K., Luwanska, A., Lassocinski, W., & Kaczmarek, Z. (2008). Estimation ofCannabis sativaL. Tissue Culture Conditions Essential for Callus Induction and Plant Regeneration. Journal of Natural Fibers, 5(3), 199-207. doi:10.1080/15440470801976045 es_ES


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