Biotecnología

 

Establishment of cell suspension cultures of Prosopis laevigata (Humb. & Bonpl. ex willd) M.C. Johnst to determine the effect of zinc on the uptake and accumulation of lead

 

Establecimiento de cultivos de células en suspensión de Prosopis laevigata (Humb. & Bonpl. ex willd) M.C. Johnst para determinar el efecto del zinc en la absorción y acumulación de plomo

 

A. Maldonado-Magaña¹, J. Orozco-Villafuerte², L. Buendía-González³*, M.E. Estrada-Zúñiga³, A. Bernabé-Antonio⁴ and F. Cruz-Sosa¹

 

¹ Departamento de Biotecnología, Universidad Autónoma Metropolitana-Iztapalapa. Av. San Rafael Atltxco No. 186, Col. Vicentina C.P. 09340, México D.F., México.

² Facultad de Química, Universidad Autónoma del Estado de México. Paseo Colón esq. Paseo Tollocan s/n, Col. Residencial Colón, C.P. 50120 Toluca, Estado de México, México.

³ Facultad de Ciencias, Universidad Autónoma del Estado de México. Campus El Cerrillo, Piedras Blancas, Carretera Toluca-Ixtlahuaca km. 15.5, C.P. 50200, Toluca, Estado de México. * Corresponding author. E-mail: lety_sax@yahoo.com.mx Tel. +52-722-296-5556, Fax+52-722-296-5554.

⁴ Centro de Investigaciones Químicas, Universidad Autónoma del Estado de Morelos. Av. Universidad 1001 Chamilpa 62210, Cuernavaca, Morelos, México.

 

Received July 15, 2013
Accepted October 144, 2013

 

Abstract

In vitro studies indicate that Prosopis laevigata can be considered a potential hyperaccumulator of lead. Likewise, lead uptake has been related to protein transporters for zinc. In this work presents a protocol for the establishment of cell suspension culture to determine the effect of zinc on the uptake and accumulation of lead. A bioassay with Pb²⁺/Zn²⁺ (0.0, 0.5, and 1.0 mM) was carried out on cell suspension cultures derived from callus induced in half-strength Murashige and Skoog (MS) medium added with 6.8 μM 2,4-dichlorophenoxyacetic acid (2,4-D) with 4.5 μM kinetin (KIN). Cells showed significant tolerance to growth (GR>60%) at all concentrations and combinations of Pb and Zn (0.0, 0.5 and 1.0 mM). When the Pb with or without Zn were added to the culture medium, the cells showed the highest accumulation efficiency for non-essential (lead) metal over essential (zinc) metal (BF values for Pb >> BF values for Zn; 2-33 times). Scanning electron micrographs evidenced the accumulation of Pb in the cells walls. These results provide insights about the tolerance and accumulation mechanisms of Pb occurring in P. laevigata.

Keywords: Prosopis laevigata, lead, zinc, competitive transport, bioaccumulation, phytoremediation.

 

Resumen

Estudios de cultivos in vitro, indican que Prosopis laevigata puede ser considerada como una especie potencialmente hiperacumuladora de plomo. En este trabajo se presenta un protocolo para el establecimiento de un cultivo de células en suspensión de P. laevigata, para determinar el efecto del Zn²⁺ sobre la absorción y acumulación de Pb²⁺. Se realizó un bioensayo con Pb²⁺/Zn²⁺ (0.0, 0.5, and 1.0 mM) en cultivos de celulas en suspension establecidos a partir de callos inducidos en cotiledones en medio Murashige & Skoog (MS) a la mitad de su concentración y suplementado con 2,4-D (6.8 μM) y KIN (4.5 μM). Las células presentaron un crecimiento relativo del 63-98% en todas las concentraciones y combinaciones de Pb y Zn (0.0, 0.5, 1.0 mM). Respecto a la acumulación, cuando el Pb fue adicionado al medio con o sin Zn, las células mostraron mayor eficiencia de acumulación para el metal no esencial (Pb) sobre el metal esencial (Zn) (valores de BF para Pb >> valores de BF para zinc; 2-33 veces). Imágenes de las células observadas con microscopía electrónica de barrido evidencian la acumulación del plomo en la pared celular. Estos resultados proporcionan información sobre los mecanismos de tolerancia y acumulación de plomo que se llevan a cabo en Prosopis laevigata.

Palabras clave: Prosopis laevigata, plomo, zinc, transporte competitivo, bioacumulación, fitorremediación.

 

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Acknowledgements

The authors wish to thank the Programa para el Mejoramiento del Profesorado for the partial financing of this project through grant PROMEP/103.5/12/3510. Also, author AM-M thanks the Consejo Nacional de Ciencia y Tecnología (CONACyT) for the scholarship received.

 

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