INDUCTION OF POLYPLOIDY: EFFECT OF COLCHICINE ON NODAL SEGMENTS OF STEVIA REBAUDIANA (BERTONI) VC. KH/IAN 142 (EIRETE)

Authors

  • Edith Alba Segovia Corrales Universidad Nacional de Asunción, CEMIT - Paraguay
  • Denice Natalie Álvarez Ramos Universidad Nacional de Asunción, CEMIT - Paraguay
  • Rosa Oviedo de Cristaldo Universidad Nacional de Asunción, DGICT - Paraguay
  • Edgar Álvarez PureCircle - Paraguay

Keywords:

Ka´a he´e, Polyploidy, Nodal cutting, Colchicine

Abstract

Stevia rebaudiana, native to Paraguay, belongs to the Asteraceae family. It is a semi-perennial plant whose main economic and social value is the production of steviosides. Stevioside (tri-glucoside) and rebaudioside A (tetra-glucoside) typically represent the majority of steviol glycosides present in Stevia rebaudiana leaves. An alternative to increase the production of these secondary metabolites and increase agronomic yield (i.e. leaf mass), may be the artificial induction of polyploidy mediated by colchicine. The objective of this study was to evaluate the use of colchicine to increase chromosome number in the species Stevia rebaudiana (Bertoni) Bertoni cv. KH/IAN 142 (Eireté). The study followed a split-plot experimental design focusing on populations of 720 nodal cutting segments (in vitro). The cuttings were subjected to three concentrations of colchicine, which were 0.01, 0.05 and 0.1 g/100 mL, at three exposure times (4 h, 8 h and 12 h), at 25 °C. As a result, the cuttings showed no changes in ploidy level, remaining diploid throughout the process. This may be due to the type of explant chosen and the exposure time to treatment for that explant. We conclude that the treatments with the different concentrations of colchicine at the exposure times used were not viable for obtaining polyploid cells.

Downloads

Download data is not yet available.

Author Biographies

Edith Alba Segovia Corrales, Universidad Nacional de Asunción, CEMIT - Paraguay

PhD. Universidad Nacional de Asunción, CEMIT. Correo electrónico: edith.segovia@cemit.una.py *autor para correspondencia.

Denice Natalie Álvarez Ramos , Universidad Nacional de Asunción, CEMIT - Paraguay

Master. Universidad Nacional de Asunción, CEMIT. Correo electrónico: dalvarezramos@gmail.com

Rosa Oviedo de Cristaldo, Universidad Nacional de Asunción, DGICT - Paraguay

PhD. Universidad Nacional de Asunción, DGICT. Correo electrónico: rosa.cristaldo@gmail.com

Edgar Álvarez, PureCircle - Paraguay

MSc. PURE CIRCLE S.A. Correo electrónico: edgar.alvarez@purecircle.com

References

Ahmar, S., Gill, R. A., Jung, K. H., Faheem, A., Qasim, M. U., Mubeen, M., & Zhou, W. (2020). Conventional and molecular techniques from simple breeding to speed breeding in crop plants: recent advances and future outlook. International Journal of Molecular Sciences, 21(7), 2590.

Bernard, F., Moghbel, N., & Hassannejad, S. (2012). Treatment of licorice seeds with colchicine: changes in seedling DNA levels and anthocyanin and glycyrrhizic acid contents of derived callus cultures. Natural Product Communications, 7(11), 1457-1460.

Britos, R., & Park, J. (2016). Stevia rebaudiana (Bertoni) Bertoni. Ed. IPTA. Paraguay. 116 pp.

Buiza, J. I., & Cequea, H. (2001). Evaluación citogenética de la generación M1V2 de tetraploides experimentales en sábila (Aloe vera L.). Revista Científica UDO Agrícola, 1(1), 29-33.

Cabrera, A. L., Holmes, W. C., & McDaniel, S. (1996). Compositae III. Flora del Paraguay, 25, 302-306.

Cabahug, R. A. M., Khanh, H. T. T. M., Lim, K. B., & Hwang, Y. J. (2021). Phenotype and ploidy evaluation of colchicine-induced Echeveria ‘Peerless’. Toxicology and Environmental Health Sciences, 13, 17-24.

Campoagropecuario.com.py (2025). Stevia: alta demanda, baja producción y nuevos mercados en la mira. https://www.campoagropecuario.com.py/notas/4232/stevia-alta-demanda-baja-produccion-y-nuevos-mercados-en-la-mira. Consultado el 08/07/2025.

CAPECO.org.py. (2025). Área de siembra, producción y rendimiento. https://capeco.org.py/area-de-siembra-produccion-y-rendimiento/. Consultado el 08/07/2025.

Caperta, A., Delgado, M., Ressurreição, F., & Meister, A. (2006). Colchicine-induced polyploidization depends on tubulin polymerization in c-metaphase cells. Protoplasma, 227, 147–153.

Castañeda-Saucedo, M. C., Tapia-Campos, E., Ramírez-Anaya, J. D. P., & Beltrán, J. (2020). Growth and development of stevia cuttings during propagation with hormones in different months of the year. Plants, 9(3), 294.

Collavino, M., De Badaro, R., & Mroginski, L. (1995). Inducción de variaciones cromosómicas por tratamiento con colchicina en diploides micropropagados de Stevia rebaudiana (Bertoni) Bertoni. (Tesis de Grado). UNAM, Corrientes, Argentina. 19 pp.

Dolinski, R., & Kowalczyk, K. (2019). Fast direct regeneration of plants from nodal explants of Stevia rebaudiana Bert. Acta Scientiarum Polonorum Hortorum Cultus, 18(5), 95-103.

Eigsti, O. J., & Dustin, P. (1955). Colchicine in Agriculture, Medicine, Biology, and Chemistry. Iowa State College Press. Iowa, U.S.A. pp. 28-30; 383.

FAO/OIEA. (2021). Manual de mejoramiento por mutación. Tercera edición. Spencer-Lopes, M.M., Forster, B.P. y Jankuloski, L. (coords.). Viena, FAO. https://doi.org/10.4060/i9285es

Feulgen, R., & Rossenbeck, H. (1924). Mikroskopisch-chemischer Nachweis einer Nucleinsäure vom Typus der Thymonucleinsäure. Hoppe-Seyler’s Zeitschrift für physiologische Chemie, 135(1-4), 203-248.

Gandhi, S. (1997). Colchicine-induced autotetraploidy in Clitoria ternatea L. Cytologia, 62(1), 13-18.

González, M. C., & Nakayama, H. D. (2015). Radioestimulación de la germinación en Stevia rebaudiana cultivar KH-IAN VC-142 (Eireté), mediante el empleo de rayos gamma 60CO. Cultivos Tropicales, 36(4), 117-119.

Gupta, S. K., & Roy, S. K. (1986). Induced colchiploidy in Nicandra physaloides. Cytologia, 51(2), 319-324.

Hamill, D. S., Smith, M. K., & Dodd, W. A. (1992). In vitro induction of banana autotetraploids by colchicine treatment of micropropagated diploids. Australian Journal of Botany, 40(6), 887-896.

Kumar, R., Jha, K. K., Sengupta, S., Misra, S., Mahto, C. S., Chakravarty, M. K., ... & Yadav, M. (2019). Effect of colchicine treatment on plant growth and floral behaviour in cape gooseberry (Physalis peruviana L.). Journal of Pharmacognosy and Phytochemistry, 8(5), 405-411.

M. de Oliveira, V., Forni-Martins, R. E., Magalhães, M. P., & Alves, N. M. (2004). Chromosomal and morphological studies of diploid and polyploidy cytotypes of Stevia rebaudiana (Bertoni) Bertoni (Eupatorieae, Asteraceae). Genetics and Molecular Biology, 27(2), 215-222.

Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15(3), 473-497.

Niazian, M., & Nalousi, A. M. (2020). Artificial polyploidy induction for improvement of ornamental and medicinal plants. Plant Cell, Tissue and Organ Culture (PCTOC), 142(3), 447-469.

Nilanthi, D., Chen, X. L., Zhao, F. C., Yang, Y. S., & Wu, H. (2009). Induction of tetraploids from petiole explants through colchicine treatments in Echinacea purpurea L. Journal of Biomedicine and Biotechnology, 2009, 1-7.

Pehlivan, E. C., & Kunter, B. (2021). Effects of colchicine on in vitro regeneration and ploidy level in grapevine (Vitis vinifera L.). Agricultural and Natural Research & Reviews, 99.

Pin, A. (2009). Plantas Medicinales del Jardín Botánico de Asunción. Asunción, Paraguay. p. 441.

Pinheiro, A. A., Pozzobon, M. T., Do Valle, C. B., Penteado, M. I. O., & Carneiro, V. T. C. (2000). Duplication of the chromosome number of diploid Brachiaria brizantha plants using colchicine. Plant Cell Reports, 19, 274–278.

Romero-Aranda, R., Bondada, B. R., Syvertsen, J. P., & Grosser, J. W. (1997). Leaf characteristics and net gas exchange of diploid and autotetraploid citrus. Annals of Botany, 79(2), 153-160.

Roychowdhury, R., & Tah, J. (2013). Mutagenesis—A potential approach for crop improvement. Crop Improvement: New Approaches and Modern Techniques, 149-187.

Salazar Mercado, S. A., Quintero Caleño, J. D., & Bustos Urbano, V. J. (2021). Impact of colchicine on leaf morphology and stomatics of Kalanchoe tubiflora (Harv.) Raym.-Hamet (Crassulaceae). Botanica Pacifica, 10(2), 1-10. DOI: 10.17581/bp.2021.10201.

Sanabria-Velazquez, A. D., Enciso-Maldonado, G. A., Maidana-Ojeda, M., Diaz-Najera, J. F., Thiessen, L. D., & Shew, H. D. (2023). Validation of standard area diagrams to estimate the severity of Septoria leaf spot on stevia in Paraguay, Mexico, and the United States. Plant Disease, 107(6), 1829-1838.

Sartor, M. E., Espinoza, F., & Quarin, C. L. (2004). Inducción de autoploidía en Paspalum plicatulum. Comunicaciones Científicas y Tecnológicas. Universidad Nacional del Nordeste, Corrientes, Argentina.

Sattler, M. C., Carvalho, C. R., & Clarindo, W. R. (2016). The polyploidy and its key role in plant breeding. Planta, 243, 281-296.

Slobodnick, A., Shah, B., Krasnokutsky, S., & Pillinger, M. H. (2018). Update on colchicine. Rheumatology, 57(suppl_1), i4-i11.

Surson, S., Sitthaphanit, S., & Wongma, N. (2015). In vivo induction of tetraploid in tangerine citrus plants (Citrus reticulata Blanco) with the use of colchicine. Pakistan Journal of Biological Sciences, 18(1), 37-41.

Villalba, B. L., & Nakayama, N. H. (2016). Efecto del estrés hídrico inducido con PEG-6000 sobre el crecimiento in vitro de plantas de Stevia rebaudiana cv. ‘KH-IAN/VC-142’. Biotecnología Vegetal, 16, 189-192.

Villalba Martínez, C. J., & Oroa Pfefferkorn, E. D. (2018). Concentración de nutrientes en tres variedades de stevia [Stevia rebaudiana (Bertoni) Bertoni] cultivadas en un Ultisol. Informaciones Agronómicas de Hispanoamérica, 29, 9-13.

Xu, B., & Li, L. (1982). Karyotype analysis of Stevia rebaudiana (Compositae). Bulletin of Jiangsu Agricultural College, 3, 17-19.

Yadav, A. K., Singh, S., Dhyani, D., & Ahuja, P. S. (2011). A review on the improvement of stevia [Stevia rebaudiana (Bertoni)]. Canadian Journal of Plant Science, 91(1), 1-27.

Yang, Y. W., Hsing, Y. I., & Chang, W. C. (1981). Clonal propagation of Stevia rebaudiana Bertoni through axillary shoot proliferation in vitro. Botanical Bulletin of Academia Sinica, 22, 57-62.

Zhang, H., An, S., Hu, J., Lin, Z., Liu, X., Bao, H., & Chen, R. (2018). Induction, identification and characterization of polyploidy in Stevia rebaudiana Bertoni. Plant Biotechnology, 35(1), 81-86.

Published

2025-12-30

Most read articles by the same author(s)