Microbial contamination during stages of in vitro propagation of Agave angustifolia Haw.

Authors

  • Fátima Manuel-Zárate Tecnológico Nacional de México-Instituto Tecnológico del Valle de Oaxaca. Ex hacienda Nazareno, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 3637836 , Tecnológico Nacional de México-Instituto Tecnológico del Valle de Oaxaca. Ex hacienda Nazareno, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 3637836
  • José Raymundo Enríquez-del Valle Tecnológico Nacional de México-Instituto Tecnológico del Valle de Oaxaca. Ex hacienda Nazareno, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 3637836 , Tecnológico Nacional de México-Instituto Tecnológico del Valle de Oaxaca. Ex hacienda Nazareno, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 3637836
  • Alfonso Vásquez-López Centro Interdisciplinario de Investigación para el Desarrollo Integral Regional-Unidad Oaxaca. Calle Hornos núm. 1003, colonia Noche Buena, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 2568667 , Centro Interdisciplinario de Investigación para el Desarrollo Integral Regional-Unidad Oaxaca. Calle Hornos núm. 1003, colonia Noche Buena, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 2568667
  • Gerardo Rodríguez-Ortiz Tecnológico Nacional de México-Instituto Tecnológico del Valle de Oaxaca. Ex hacienda Nazareno, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 3637836 , Tecnológico Nacional de México-Instituto Tecnológico del Valle de Oaxaca. Ex hacienda Nazareno, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 3637836
  • Vicente Arturo Velasco-Velasco Tecnológico Nacional de México-Instituto Tecnológico del Valle de Oaxaca. Ex hacienda Nazareno, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 3637836 , Tecnológico Nacional de México-Instituto Tecnológico del Valle de Oaxaca. Ex hacienda Nazareno, Santa Cruz Xoxocotlán, Oaxaca, México. CP. 71230. Tel. 951 3637836

DOI:

https://doi.org/10.29312/remexca.v17i2.4251

Keywords:

antibiotics, microbial contamination, microorganisms, organogenesis

Abstract

When Agave angustifolia stem tissues are established in vitro, microbial contamination frequently occurs; therefore, the use of antibiotics to solve this problem was evaluated. The purpose of this work was to determine the percentage of stem tissues that had an aseptic establishment and exhibited morphogenetic response in three situations: 1) during in vitro establishment (stage I); 2) in explants in which organogenesis with shoot formation occurred in stage I but were contaminated with bacteria, and these were treated with antibiotics to eliminate contamination; and 3) in aseptic adventitious shoots obtained in stage I and transferred to propagule multiplication medium (stage II). The percentages of aseptic cultures, viable tissues, and organogenic response were evaluated. The results showed that only between 3.68 and 8.73% of stem tissues were aseptic during stage I. However, by applying antibiotics to shoots already formed but contaminated with bacteria at this stage, asepsis was recovered in 52.5% of the cultures. On the other hand, adventitious shoots that were aseptic from stage I and transferred to the multiplication medium (stage II) maintained asepsis at 89.88%; all were viable and showed formation of new shoots. These results indicate that antibiotic treatment in already induced stem tissues is an effective strategy to rescue contaminated cultures, and that transferring aseptic shoots to the multiplication stage allows maintaining aseptic cultures with morphogenetic response.

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References

Abreu, E.; Sosa-Castillo, M.; Ascunce-Sol G. y González, G. 2016. Efecto de antibióticos en la propagación in vitro de Agave fourcroydes Lem. Biotecnología Vegetal. 16(1):31-36. https://revista.ibp.co.cu/index.php/BV/article/view/507.

Altan, F.; Bürün, B. and Sahin, N. 2010. Fungal contaminants observed during micropropagation of Lilium candidum L. and the effect of chemotherapeutic substances applied after sterilization. African Journal of Biotechnology. 9(7):991-995.

Azofeifa, A. 2009. Problemas de oxidación y oscurecimiento de explantes cultivados in vitro. Agronomía Mesoamericana. 20(1):153-175. https://archivo.revistas.ucr.ac.cr/index.php/agromeso/article/view/4990/4799.

Cruz-Hernández, H.; Enríquez-Valle, J. R.; Velasco-Velasco, V. A.; Ruiz-Luna, J.; Campos-Ángeles, G. V. y Aquino, D. E. 2013. Nutrimentos y carbohidratos en plantas de Agave angustifolia Haw. y Agave karwinskii Zucc. Revista Mexicana de Ciencias Agrícolas. 6(4):1161-1173. https://www.scielo.org.mx/pdf/remexca/v4nspe6/v4spe6a8.pdf.

Debergh, P. C. and Maene, L. J. 1981. A Scheme for commercial propagation of ornamental plants by tissue culture. Scientia Horticulturae. 14(4):335-345. https://doi.org/10.1016/0304-4238(81)90047-9.

DGSIAP. 2024. Dirección General del Servicio de Información Agroalimentaria y Pesquera. Anuario estadístico de la producción agrícola. https://nube.agricultura.gob.mx/cierre-agricola/.

Domínguez-Rosales, M. S.; González-Jiménez, M. L.; Rosales-Gómez, C.; Quiñones-Valles, C.; Delgadillo, D. S.; Mireles-Ordaz, S. J. y Pérez-Molphe, B. E. 2008. El cultivo in vitro como herramienta para el aprovechamiento, mejoramiento y conservación de especies del género agave. Investigación y Ciencia. Universidad Autónoma de Aguascalientes. 41:53-62. https://dialnet.unirioja.es/servlet/articulo?codigo=6104556.

Enjalric, F.; Carron, M. P. and Lardet, L. 1988. Contamination of primary cultures in tropical areas: the case of Hevea brasiliensis. Acta Horticulturae. 225:57-66. https://doi.org/10.17660/ActaHortic.1988.225.6.

Enríquez-Valle, J. R. 2008. La propagación y crecimiento de agaves. Fundación Produce Oaxaca, AC. Instituto Tecnológico del Valle de Oaxaca (ITVO). Oaxaca. México. ISBN: 978-607-00. 46 p.

Fortes, A. M. and Pais, M. S. 2000. Organogenesis from intemode-derived nodules of Humulus lupulus var. Nugget (Cannabinaceae): histoligical studies and changes in the starch content. American Journal of Botanic. 87(7):971-979. https://doi.org/10.2307/2656996.

George, E. F. and Debergh, P. C. 2008. Micropropagation: uses and methods. In: George, E. F.; Hall, M. and De Klerk, G. Plant propagation by tissue culture. 3er Ed. The Bacground. Springer. 29-64 pp.

González-González, L. R.; García-Pérez, M. I. B.; Gutiérrez, L. K. y García, A. 2007. Obtención de azúcares fermentables a partir de inulinasas inmovilizadas por el método del sol-gel. Rev. Cienc. Tecnol. 6(6):106-111.

Jarquín-Rosales, D.; Enríquez-Valle, J. R.; Alpuche-Osorno, J. J.; Rodríguez-Ortiz, G.; Martin, M. P. and Campos-Ángeles, G. V. 2022. The effects of fertirrigation and Azospirillum brasilense inoculation on photosynthetic compounds of Agave angustifolia. Australian Journal of Crop Science. 16:162-168. Doi: http://doi.org/10.21475/ajcs.22.16.01.p3280.

Leifert, C. and Cassells, A. C. 2001. Microbial hazards in plant tissue and cell cultures. In Vitro Cellular & Developmental Biology Plant. 37(2):133-138. https://doi.org/10.1007/s11627-001-0025-y.

Leifert, C. and Waites, W. M. 1992. Bacterial growth in plant tissue culture media. Journal of Applied Bacteriology. 72:460-466. https://doi.org/10.1111/j.1365-2672.1992.tb01859.x.

Leifert, C.; Ritchie, J. Y. and Waites, W. M. 1991. Contaminants of plant tissue cultures. World Journal of Microbiology and Biotechnology. 37(2):133-138. https://doi.org/10.1007/BF00303371.

López-Acevedo, L.; Merino-Pérez, Y. E.; Enríquez-Valle J. R.; Rodríguez-Ortiz, G. and Lagunas-Sánchez, Z. C. 2018. Organogénesis in vitro en tejidos de tallo de Agave marmorata y Agave angustifolia. Revista Mexicana de Agroecosistemas. 5(2):20-27. https://revistaremaeitvo.mx/index.php/remae/article/view/165.

Miguel-Luna, M. E.; Enríquez-Valle, J. R.; Velasco-Velasco, V. A.; Villegas-Aparicio, Y.; Carrillo-Rodríguez, J. C. y Rodríguez-Ortíz, G. 2013. Composición del medio de cultivo y la incubación para enraizar brotes de Agave. Revista Mexicana de Ciencias Agrícolas. 4(6):1151-1159. https://www.scielo.org.mx/pdf/remexca/v4nspe6/v4spe6a7.pdf.

Millán-Soto, G.; Gutiérrez, A.; Esqueda, M.; Gardea, A.; Tiznado, M. and Orozco, J. A. 2016. Respiratory metabolism of Agave angustifolia Haw. Clonal lines at different temperatures. Plant Cell Tissue Organ Cult. 125(1):71-80. https://doi.org/10.1007/s11240-015-0930-0.

Morales-González, B. and Núñez-Palenius, H. G. 2015. Micropropagación de Agave tequilana Weber variedad Azul ‘El coronel’ en un sistema de inmersión temporal (SIT). Jóvenes en la Ciencia. 1(2):77 82. http://repositorio.ugto.mx/handle/20.500.12059/2307.

Murashige, T. and Skoog, F. 1962. A Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. 15(3):473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x.

Obledo-Vázquez, E. N.; Flores-Verduzco, N. y Cervantes-Martínez, J. 2004. Detección del efecto de un extracto vegetal antimicrobiano sobre plantas de agave (Agave tequilana Weber var. azul) Cultivadas in vitro utilizando fluorescencia inducida por láser (LIF). Revista Mexicana de Fitopatología. 22(3):328-332. https://www.redalyc.org/pdf/612/61222302.pdf.

Pancaningtyas, S. 2015. Study on the presence and influence of phenolic compounds in callogenesis and somatic embryo development of cocoa (Theobroma cacao L.). Pelita Perkebunan. 31(1):14-20.

Pérez-Pazos, J.; Rosero, A.; Cardinale, M. and Gámez, R. 2023. Development of control strategies for bacteria and fungi associated with a micropropagated new cultivar of orange-fleshed sweet potato (Ipomoea batatas cv. Agrosavia-Aurora). Hortic. Environ. Biotechnol. 64:859-875. https://doi.org/10.1007/s13580-023-00521-2.

Ríos-Ramírez, S. C.; Enríquez- Valle, J. R.; Rodríguez-Ortiz, G.; Ruíz-Luna, J. and Velasco-Velasco, V. A. 2018. In vitro formation of adventitious shoots on culinary tissue of physiologically contrasting Agave angustifolia plants. Emirates Journal of Food and Agriculture. 30(1):285-294.

Robert, M. L.; Herrera, J. L.; Chan, J. L. and Contreras, F. 1992. Micropropagation of Agave spp. In: Bajaj, Y. P. S. Ed. Biotechnology in agriculture and forestry. 19. Springer-Verlag. 306-329 pp. https://doi.org/10.1007/978-3-662-07770-2-19.

Sánchez, A.; Coronel-Lara, Z.; Gutiérrez, A.; Vargas, G.; Coronado, M. L. y Esqueda, M. 2020. Aclimatación y trasplante de vitro plantas de Agave angustifolia Haw. en condiciones silvestres. Revista Mexicana Ciencias Agrícolas. 11(7):1593-1605. https://doi.org/10.29312/remexca.v11i7.2403.

Sánchez-Cuevas, J. L. y Salaverria, J. 2004. Control de la oxidación y la contaminación en el cultivo in vitro de fresa (Fragaria x ananassa Duch. Revista Científica UDO Agrícola. 4(1):21- 26. https://dialnet.unirioja.es/servlet/articulo?codigo=2221549.

SAS/ETS® 9.4. 2019. SAS Institute Inc. Cary, North Carolina, USA.

SIAP. 2019. Servicio de Información Agroalimentaria y Pesquera. Anuario Estadístico de la Producción Agrícola. https://nube.siap.gob.mx/cierreagricola.

Steiner, A. A. 1961. A universal method for preparing nutrient solutions of a certain desired composition. Plant Soil. 15:134-154.

Zeng, F.; Zhang, X.; Cheng, L.; Hu, L.; Zhu, L.; Cao, J. and Guo, X. 2007. A draft gene regulatory network for celular totipotency reprogramming during plant somatic embryogenesis. Genomics. 90:620-628.

Published

2026-04-26

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Articles

How to Cite

Manuel-Zárate, Fátima, José Raymundo Enríquez-del Valle, Alfonso Vásquez-López, Gerardo Rodríguez-Ortiz, and Vicente Arturo Velasco-Velasco. 2026. “Microbial Contamination During Stages of in Vitro Propagation of Agave Angustifolia Haw”. Revista Mexicana De Ciencias Agrícolas 17 (2): e4251. https://doi.org/10.29312/remexca.v17i2.4251.

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