Production of callus and roots from lateral meristems of Loeselia mexicana

Lizeth Aguirre-Alberto, Maria de Lourdes Martinez-Cardenas


Background: Loeselia mexicana, known as Mexican false calico, or espinosilla in Spanish, is a widely distributed secondary forest plant with a significant number of medicinal and cosmetic uses. This species is threatened by the lack of regulation over collection methods and changes in land use. In vitro culture could be used to preserve the species by shoot induction, callus production and cell-suspension to obtain secondary metabolites; this would reduce the need to affect wild populations.

Hypothesis: A combination of cytokinins and auxins can induce structural development in the plant, promoting the formation of shoots, roots or callus in vitro. By applying this combination to L. mexicana stem segments, we expected to observe new shoots or callus.

Study site and dates: “El Teuhtli” volcano, Xochimilco; from June 2015 to February 2016.

Methods: Distal stems cuttings were used as explants. They were disinfected with 1 % soap, 0.6 sodium hypochlorite and 70 % ethanol, and rinsed with distilled water. Two different times of disinfection with ethanol were tested. The distal stem segments were then planted in solid MS medium with 3, 5 or 7 mg L-1 KIN combined with 3 mg L-1 NAA, and 2 % AC.

Results: A favorable response was observed in the treatment with 5 mg L-1 KIN and 3 mg L-1 NAA, which produced green callus with root in five weeks. Furthermore, a lower explant mortality rate was achieved, 46.66 % in three weeks, with a shorter disinfection time.

Conclusions: Disinfection time is important for this species, and callus production is possible.


callus; espinosilla; medicinal; meristems; organogenesis

Full Text:



Arizaga S. 2007. Estudio y Colección Viva de Plantas Medicinales Nativas y Formación de un Banco de Germoplasma del Estado de Michoacán. Informe final. UNAM-Campus Morelia: Centro de Investigaciones en Ecosistemas.

Azofeifa A. 2009. Problemas de oxidación y oscurecimiento de explantes cultivados in vitro. Agronomía Mesoamericana 20: 153-175.

Belukurava V, Kuchuk N. 2014. In vitro Bank and Seed Collection of Wild-Growing Plants as a Tool for Plant Conservation and Utilization in Biotechnological Studies. In: Behl RK, Arseniuk E, eds. Biotechnology and Plant Breeding Perspectives. India: AEROBIOS International, 119-232. ISBN-13: 978-9381191040

BDMTM [Biblioteca Digital de la Medicina Tradicional Mexicana]. 2009. Atlas de las plantas de la Medicina Tradicional Mexicana: Espinosilla. Universidad Nacional Autónoma de México. (accessed June 25, 2018)

CONABIO [Comisión Nacional para el uso de la Biodiversidad]. 2009. Polemoniaceae, Loeselia mexicana. (accessed February 12, 2018).

Cardoza V. 2008. Tissue Culture: The Manipulation of Plant Development. In: Stewart CN, ed. Plant Biotechnology and Genetics: Principles, Techniques, and Applications. USA: John Wiley and Sons, 113-134. ISBN: 9780470043813; DOI:10.1002/9780470282014

Çördük N, Aki C. 2011. Inhibition of browning problem during micropropagation of Sideritis trojana bornm., an endemic medicinal herb of Turkey. Romanian Biotechnological Letters 16: 6760-6765.

Dong Y, Fu C, Su P, Xu X, Yuan J, Wang S, Zhang M, Zhao C, Yu L. 2016. Mechanisms and effective control of physiological browning phenomena in plant cell cultures. Physiologia Plantarum 156: 13-28. DOI: 10.1111/ppl.12382

FAO [Food and Agriculture Organization of the United Nations]. 2002. Estado de la Información Forestal en México. Vol. 11. Información para el Desarrollo Forestal Sostenible, PROYECTO GCP/RLA/133/EC, Chile, 179-279.

Gamborg O, Phillips G, eds. 1995. Plant Cell, Tissue and Organ Culture: Fundamental Methods. Germany: Springer Berlin Heidelberg. DOI: 10.1007/978-3-642-79048-5; ISBN: 978-3-642-48974-7

Guzmán-Gutiérrez L, Reyes-Chilpa R, Bonilla-Jaime H. 2014. Medicinal plants for the treatment of “nervios”, anxiety, and depression in Mexican Traditional Medicine (Review). Brazilian Journal of Pharmacognosy 24: 591-608. DOI: 10.1016/j.bjp.2014.10.007

Herrera-Ruiz M, González-Carranza A, Zamilpa A, Jiménez-Ferrer E, Huerta-Reyes M, Navarro-García VM. 2011. The standardized extract of Loeselia mexicana possesses anxiolytic activity through the α-amino butyric acid mechanism. Journal of Ethnopharmacology 138: 262-267. DOI: 10.1016/j.jep.2011.09.010

Iliev I, Gajdosová A, Libiaková G, Jain SM. 2010. Plant Micropropagation. In: Davey MR, Anthony P, eds. Plant Cell Culture: Essential Methods. UK: John Wiley & Sons, 1-23. ISBN: 9780470686485; DOI: 10.1002/9780470686522

Jin L, Yang Y, Gao W, Gong M, Wang J, Anderson NO, He M. 2017. Establishment of Callus Induction and Cell Suspension Cultures of Dendrathema Indicum var. Aromaticum a Scented Chrysanthemum. Journal of Plant Studies 6: 38-44. DOI: 10.5539/jps.v6n2p38

Loyola-Vargas V, Vázquez-Flota F, eds. 2006. Plant Cell Culture Protocols. USA: Humana Press. ISBN 978-1-58829-547-7; DOI: 10.1385/1592599591

Muñetón-Pérez P. 2009. Plantas medicinales: un complemento vital para la salud de los mexicanos. Entrevista con el Dr. Erick Estrada Lugo. Revista Digital Universitaria 10: 1-9.

Murashige T, Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Biologia Plantarum 15: 473-497.

Navarro-García VM, Herrera-Ruiz M, Rojas G, Zepeda LG. 2007. Coumarin Derivatives from Loeselia mexicana. Determination of the Anxiolytic Effect of Daphnoretin on Elevated Plus-maze. Journal of the Mexican Chemical Society 51: 193-197.

Navarro-García V, Rojas G, Avilés M, Fuentes M, Zepeda G. 2011. In vitro antifungal activity of coumarin extracted from Loeselia mexicana Brand. Mycoses 54: e569-e571. DOI: 10.1111/j.1439-0507.2010.01993.x

Olmos S, Luciani G, Galdeano E. 2010. Métodos de propagación y conservación del germoplasma: Micropopagación. In: Levitus G, Echenique V, Rubinstein C, Hoop E, Mroginski L, eds. Biotecnología y Mejoramiento Vegetal II. Argentina: Instituto Nacional de Tecnología Agropecuaria (INTA), 353-362.

Palombo EA. 2006. Phytochemicals from Traditional Medicinal Plants used in the Treatment of Diarrhea: Modes of Action and Effects on Intestinal Function (Review). Phytotherapy Research 20: 717-724. DOI: 10.1002/ptr.1907

Pérez GS, Pérez GC, Zavala MA. 2005. A study of the antidiarrheal properties of Loeselia mexicana on mice and rats. Phytomedicine 12: 670-674.

Razavi SM, Arshneshin H, Ghasemian A. 2017. In vitro callus induction and isolation of volatile compounds in callus culture of Lallemantia iberica (M. Bieb.) Fisch. & C. A. Mey. Journal of Plant Process and Function 5: 65-68.

RECNAT [Recursos Naturales]. 1997. Norma Oficial Mexicana NOM-005-RECNAT-1997 que establece los procedimientos, criterios y especificaciones para realizar el aprovechamiento, transporte y almacenamiento de corteza, tallos y plantas completas de vegetación forestal. México: Diario Oficial de la Federación. 23/04/2003.

Reyes-Santiago PJ, Islas-Luna MA, González-Zorzano O, Carrillo-Reyes P, Vergara-Silva FR, Brachet-Ize CP. 2011. Echeveria: Manual del Perfil Diagnóstico del Género Echeveria en México. Mexico: Universidad Autónoma de Chapingo. ISBN: 978-607-12-0218-5

Roca W, Mroginski L. 1991. Cultivo de Tejidos en la Agricultura: Fundamentos y Aplicaciones. Colombia: Centro Internacional de Agricultura Tropical. ISBN: 958-9183-15-8

Rojas A, Bah M, Rojas J I, Serrano V, Pacheco S. 1999. Spasmolytic activity of some plants used by the Otomi Indians of Querétaro (México) for the treatment of gastrointestinal disorders. Phytomedicine 6: 367-371. DOI: 10.1016/S0944-7113(99)80061-0

Rout G, Samantaray S, Das P. 2000. In vitro manipulation and propagation of medicinal plants Biotechnology Advances 18: 91-120. DOI: 10.1016/S0734-9750(99)00026-9

Rzedowski J, Calderón de Rzedowski G. 1995. Flora del bajío y de regiones adyacentes: Polemoniaceae. Pátzcuaro, Michoacán: Instituto de Ecología, A.C.-Centro Regional del Bajío 33: 1-41

Salud-Pérez S, Cuauhtémoc-Pérez G, Zavala MA. 2005. A study of the antidiarrheal properties of Loeselia mexicana on mice and rats. Phytomedicine 12: 670-674.

SEDEREC [Secretaría de Desarrollo Rural y Equidad para las Comunidades]. 2012. Programa para la recuperación de la medicina tradicional y la herbolaria en la Ciudad de México. Tomo II. Gaceta Oficial del Distrito Federal, 106-115.

Tang K, Zhang L, Chen J, Xiao Y, Chen W, Sun X. 2010. Secondary Products. In: Davey MR, Anthony P, eds. Plant Cell Culture: Essential Methods. UK: John Wiley & Sons, 297-315. Print ISBN: 9780470686485; DOI: 10.1002/9780470686522

Trejo-Tapia G, R-Romero G, L-Laredo AR, B-Torres K, Zamilpa A. 2012. In vitro Organ Cultures of the Cancer Herb Castilleja tenuiflora Benth. as Potential Sources of Iridoids and Antioxidant Compounds. In: Orhan I, ed. Biotechnological Production of Plant Secondary Metabolites. Turkey: Bentham Science Publishers, 87-106. DOI: 10.2174/97816080511441120101; ISBN: 978-1-60805-410-7

Wang RF, Huang FL, Zhang J, Zhang QY, Sun LN, Song XS. 2016. Establishment of a high-frequency regeneration system in Cerasus humilis, an important economic shrub. Journal of Forest Research 21: 244-250. DOI: 10.1007/s10310-016-0535-4



  • There are currently no refbacks.

ISSN: 2007-4476
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.