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American Journal of Potato Research

, Volume 92, Issue 1, pp 128–137 | Cite as

Low-Cost Potato Tissue Culture with Microwave and Bleach Media Preparation and Sterilization

  • Brooke N. WeberEmail author
  • R. Andrews Witherell
  • Amy O. Charkowski
Article

Abstract

Labor and equipment costs are the main expenses in potato micropropagation. To determine if we could reduce costs associated with media sterilization, a disinfectant, sodium hypochlorite (NaOCl), in combination with microwave heating, were assayed as media sterilants. Incorporating a common 5 % NaOCl household beach at a concentration of 9 ppm (active chlorine) in media sterilized with an autoclave or microwave oven controlled microorganism growth and maintained plantlet growth performance. Non-sterile 473 ml (16 oz.) clear deli containers were selected as an inexpensive replacement for traditional culture vessels and were effectively sterilized with a 50 ppm (active chlorine) NaOCl solution. Reuse of the non-sterile clear deli containers and alternate media water sources were also tested but this decreased plantlet growth performance. Comparison of a controlled growth chamber and ambient laboratory conditions was also investigated. Microorganism growth was significantly less in a controlled growth chamber (5 %) as compared to uncontrolled conditions (26–36 %).

Keywords

Sodium hypochlorite Clear deli container Contamination Micropropagation Plantlets 

Resumen

Los costos de mano de obra y de equipo son los principales gastos en la micropropagación de papa. Con el fin de determinar si podríamos reducir los costos asociados con la esterilización del medio, se probó un desinfestante, hipoclorito de sodio (NaOCl), en combinación con calentamiento en el horno de microondas, como esterilizantes del medio. La incorporación de blanqueador doméstico de NaOCl común al 5 % a una concentración de 9 ppm (cloro activo), en medio esterilizado en una autoclave o en horno de microondas, controló el crecimiento de microorganismos y mantuvo el comportamiento del crecimiento de la plántula. Se seleccionaron recipientes transparentes no estériles de alimentos de 473 ml (16 oz.), como un reemplazo económico a los recipientes de cultivo tradicionales y se esterilizaron efectivamente con 50 ppm (cloro activo) de una solución de NaOCl. También se probó el re-uso de los recipientes transparentes no estériles y de fuentes alternativas de agua para el medio, pero esto disminuyó el buen comportamiento del crecimiento de la plántula. También se investigó la comparación de una cámara de crecimiento controlada con las condiciones ambientales de laboratorio. El crecimiento de los microorganismos fue significativamente menor en la cámara de crecimiento controlado (5 %) al compararlo con las condiciones no controladas (26–36 %).

Notes

Acknowledgments

Non-sterile clear deli containers were generously donated by M. Olson (Placon, Madison WI). The active chlorine level in tap water was kindly provided by City of Madison Water Utilities (Madison, WI). We thank N. Keuler (University of Wisconsin-Madison) for statistical advice and the following University of Wisconsin undergraduate student hourlies: E. Hutchins, E. Schmid, J. Wochos, L. Jenison, M. Moritz and R. Shefchek (University of Wisconsin) for their dedication to this project.

Supplementary material

12230_2014_9423_MOESM1_ESM.docx (67 kb)
ESM 1 (DOCX 67 kb)

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Copyright information

© The Potato Association of America 2014

Authors and Affiliations

  • Brooke N. Weber
    • 1
    Email author
  • R. Andrews Witherell
    • 1
  • Amy O. Charkowski
    • 1
  1. 1.Wisconsin Seed Potato Certification Program Tissue Culture Laboratory Department of Plant PathologyUniversity of Wisconsin-MadisonMadisonUSA

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