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Influence of Pulsed Electric Field Protocols on the Reversible Permeabilization of Rucola Leaves

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Abstract

Reversible electropermeabilization of plant tissues with heterogeneous structure represents a technological challenge as the response of the different structures within the same specimen to the application of electric field may differ due to different cell sizes, extracellular space configurations, and electrical properties. The influence of five different pulsed electric field protocols with different pulse polarity, number of pulses (25, 50, 75, 100, 250, and 500), and intervals between pulses (no intervals and 1- and 2-ms intervals) on the reversible permeabilization of rucola (Eruca sativa) leaves was investigated. The electric field intensity was 600 V/cm. Electrical resistance of the bulk tissue was measured before and after electroporation, and propidium iodide was used to analyze the electroporation at the surface of the leaf. Leaf viability was assessed from survival in storage, and cell viability was investigated with fluorescein diacetate. Results indicate that the viability of the leaves could not be predicted by measurements of electrical resistance or permeabilization levels of the leaf surface. Higher survival rate was demonstrated when applying bipolar pulses compared with monopolar pulses, but the latter proved to be more effective than bipolar pulses for permeabilizing the surface of the leaves. Longer intervals between bipolar pulses resulted in increased viability preservation, while the number of electroporated cells on the leaf surface was comparable for all tested protocols.

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References

  • Asavasanti, S., Ristenpart, W., Stroeve, P., & Barrett, D. M. (2011). Permeabilization of plant tissues by monopolar pulsed electric fields: effect of frequency. Journal of Food Science, 76(1), E98–E111.

    Article  CAS  Google Scholar 

  • Ben Ammar, J., Lanoisellé, J.-L., Lebovka, N., Van Hecke, E., & Vorobiev, E. (2011). Impact of a pulsed electric field on damage of plant tissues: effects of cell size and tissue electrical conductivity. Journal of Food Science, 76(1), E90–E97.

    Article  CAS  Google Scholar 

  • Benz, R., & Zimmermann, U. (1981). The resealing process of lipid bilayers after reversible electrical breakdown. Biochimica et Biophysica Acta, 640(1), 169–178.

    Article  CAS  Google Scholar 

  • Berger, F., Linstead, P., Dolan, L., & Haseloff, J. (1998). Stomata patterning on the hypocotyl of Arabidopsis thaliana is controlled by genes involved in the control of root epidermis patterning. Developmental Biology, 194(2), 226–234.

    Article  CAS  Google Scholar 

  • Berridge, M. J., Bootman, M. D., & Lipp, P. (1998). Calcium—a life and death signal. Nature, 395(6703), 645–648.

    Article  CAS  Google Scholar 

  • Beveridge, J. R., Wall, K. A., MacGregor, S. J., Anderson, J. G., & Rowan, N. (2003). Pulsed electric field in inactivation of spoilage microorganisms in alcoholic beverages and the influence of pulse profile. Proceedings of the IEEE, 92(7), 1138–1143.

    Article  Google Scholar 

  • Beveridge, J. R., MacGregor, S. J., Anderson, J. G., & Fouracre, R. A. (2005). The influence of pulse duration on the inactivation of bacteria using monopolar and bipolar profile pulsed electric fields. IEEE Transactions on Plasma Science, 33(4), 1287–1293.

    Article  Google Scholar 

  • Bilska, A. O., DeBruin, K. A., & Krassowska, W. (2000). Theoretical modeling of the effects of shock duration, frequency, and strength on the degree of electroporation. Bioelectrochemistry, 51(2), 133–143.

    Article  CAS  Google Scholar 

  • Bouzrara, H., & Vorobiev, E. (2000). Beet juice extraction by pressing and pulsed electric fields. International Sugar Journal, 102(1216), 194–200.

    CAS  Google Scholar 

  • Buescher, E. S., & Schoenbach, K. H. (2003). Effects of submicrosecond, high intensity pulsed electric fields on living cells—intracellular electromanipulation. IEEE Transactions on Dielectrics and Electrical Insulation, 10(5), 788–794.

    Article  Google Scholar 

  • Chalermchat Y. (2005). Effects of pulsed electric fields on plant tissue. Ph.D. thesis, Department of Food Technology, Engineering and Nutrition, Lund University, Lund.

  • Chalermchat, Y., Malangone, L., & Dejmek, P. (2010). Electropermeabilization of apple tissue: effect of cell size, cell size distribution and cell orientation. Biosystems Engineering, 105(3), 357–366.

    Article  Google Scholar 

  • Charlene, A. J., Atsushi, H., & Wildman, S. G. (1978). Evidence that the amount of chloroplast DNA exceeds that of nuclear DNA in mature leaves. The Journal of Cell Biology, 79(3), 631–636.

    Article  Google Scholar 

  • Coley, P. D. (1980). Effects of leaf age and plant life history patterns on herbivory. Nature, 284(5756), 545–546.

    Article  Google Scholar 

  • Esser, A. T., Smith, K. C., Gowrishankar, T. R., Vasilkoski, Z., & Weaver, J. C. (2010). Mechanisms for the intracellular manipulation of organelles by conventional electroporation. Biophysical Journal, 98(11), 2506–2514.

    Article  CAS  Google Scholar 

  • Evrendilek, G. A., & Zhang, Q. H. (2005). Effects of pulse polarity and pulse delaying time on pulsed electric fields-induced pasteurization of E. coli O157:H7. Journal of Food Engineering, 68(2), 271–276.

    Article  Google Scholar 

  • Evrendilek, G. A., Zhang, Q. H., & Richter, E. R. (1999). Inactivation of Escherichia coli O157:H7 and Escherichia coli 8739 in apple juice by pulsed electric fields. Journal of Food Protection, 62(7), 793–796.

    CAS  Google Scholar 

  • Gabriel, B., & Teissié, J. (1994). Generation of reactive-oxygen species induced by electropermeabilization of Chinese hamster ovary cells and their consequence on cell viability. European Journal of Biochemistry, 223(1), 25–33.

    Article  CAS  Google Scholar 

  • Gabriel, B., & Teissié, J. (1997). Direct observation in the millisecond time range of fluorescent molecule asymmetrical interaction with the electropermeabilized cell membrane. Biophysical Journal, 73(5), 2630–2637.

    Article  CAS  Google Scholar 

  • Gabriel, B., & Teissié, J. (1999). Time courses of mammalian cell electropermeabilization observed by millisecond imaging of membrane property changes during the pulse. Biophysical Journal, 76(4), 2158–2165.

    Article  CAS  Google Scholar 

  • Glaser, R., Leikin, S., Chernomordik, L., Pastushenko, V., & Sokirko, A. (1988). Reversible electrical breakdown of lipid bilayers: formation and evolution of pores. Biochimica et Biophysica Acta, 940(2), 275–287.

    Article  CAS  Google Scholar 

  • Golzio, M., Teissié, J., & Rols, M.-P. (2002). Direct visualization at the single-cell level of electrically mediated gene delivery. Proceedings of the National Academy of Sciences, 99(3), 1292–1297.

    Article  CAS  Google Scholar 

  • Gómez Galindo, F., Wadsö, L., Vicente, A., & Dejmek, P. (2008). Exploring metabolic responses of potato tissue induced by electric pulses. Food Biophysics, 3(4), 352–360.

    Article  Google Scholar 

  • Hapala, I. (1997). Breaking the barrier: methods for reversible permeabilization of cellular membranes. Critical Reviews in Biotechnology, 17(2), 105–122.

    Article  CAS  Google Scholar 

  • Hill, R. M., Dissado, L. A., & Pathmanathan, K. (1987). The low-frequency dielectric properties of leaves. Journal of Biological Physics, 15(1), 2–16.

    Article  Google Scholar 

  • Khan, O. G. M. & El-Hag, A. H. (2011). Biological cell electroporation using nanosecond electrical pulses. In: Proceedings of 1st Middle East Conference on Biomedical Engineering (MECBME), 21–24 February 2011, Sharjah.

  • Kotnik, T., Mir, L., Flisar, K., Puc, M., & Miklavcic, D. (2001). Cell membrane electropermeabilization by symmetrical bipolar rectangular pulses: part I. Increased efficiency of permeabilization. Bioelectrochemisty, 54(1), 83–90.

    Article  CAS  Google Scholar 

  • Lebar, A. M., & Miklavčič, D. (2001). Cell electropermeabilization to small molecules in vitro: control by pulse parameters. Radiology and Oncology, 35(3), 193–202.

    Google Scholar 

  • Mañas, P., Barsotti, L., & Cheftel, J. C. (2001). Microbial inactivation by pulsed electric fields in a batch treatment chamber: effects of some electrical parameters and food constituents. Innovative Food Science and Emerging Technologies, 2(4), 239–249.

    Article  Google Scholar 

  • Martín-Belloso, O., Vega-Mercado, H., Qin, B. L., Chang, F. J., Barbosa-Canovas, G. V., & Swanson, B. C. (1997). Inactivation of Escherichia coli suspended in liquid egg using pulsed electric fields. Journal of Food Processing and Preservation, 21(3), 193–208.

    Article  Google Scholar 

  • Neumann, E., & Rosenhec, K. (1972). Permeability changes induced by electric impulses in vesicular membranes. The Journal of Membrane Biology, 10(1), 279–290.

    Article  CAS  Google Scholar 

  • Neumann, E., Schaefer-Ridder, M., Wang, Y., & Hofschneider, P. (1982). Gene transfer into mouse lyoma cells by electroporation in high electric fields. The EMBO Journal, 1(7), 841–845.

    CAS  Google Scholar 

  • Pakhomov, A. G., Miklavčič, D., & Markov, M. S. (2010). Advanced electroporation techniques in biology and medicine. New York: CRC.

    Google Scholar 

  • Phillips, M., Maor, E., & Rubinsky, B. (2011). Principles of tissue engineering with nonthermal irreversible electroporation. Journal of Heat Transfer, 133(1).

  • Phoon, P. Y., Galindo, F. G., Vicente, A., & Dejmek, P. (2008). Pulsed electric field in combination with vacuum impregnation with trehalose improves the freezing tolerance of spinach leaves. Journal of Food Engineering, 88(1), 144–148.

    Article  CAS  Google Scholar 

  • Psaras, G. K., & Rhizopoulou, S. (1995). Mesophyll structure during leaf development in Ballota acetabulosa. The New Phytologist, 131(3), 303–309.

    Article  Google Scholar 

  • Qin, B.-L., Zhang, Q., Barbosa-Canovas, G. V., Swanson, B. G., & Pedrow, P. D. (1994). Inactivation of microorganisms by pulsed electric fields of different voltage waveforms. IEEE Transactions on Dielectrics and Electrical Insulation, 1(6), 1047–1057.

    Article  Google Scholar 

  • Qin, B. L., Pothakamury, U. R., Barbosa-Cánovas, G. V., & Swanson, B. G. (1996). Nonthermal pasteurization of liquid foods using high-intensity pulsed electric fields. Critical Reviews in Food Science and Nutrition, 36(6), 603–627.

    Article  CAS  Google Scholar 

  • Qin, B.-L., Barbosa-Canovas, G. V., Swanson, B. G., Pedrow, P. D., & Olsen, R. G. (1998). Inactivating microorganisms using a pulsed electric field continuous treatment system. IEEE Transactions on Industry Applications, 34(1), 43–50.

    Article  Google Scholar 

  • Riederer, M., & Schönherr, J. (1988). Development of plant cuticles: fine structure and cutin composition of Clivia miniata Reg. leaves. Planta, 174(1), 127–138.

    Article  CAS  Google Scholar 

  • Rols, M.-P., & Teissié, J. (1998). Electropermeabilization of mammalian cells to macromolecules: control by pulse duration. Biophysical Journal, 75(3), 1415–1423.

    Article  CAS  Google Scholar 

  • Rowan, N. J., MacGregor, S. J., Anderson, J. G., Fouracre, R. A., & Farish, O. (2000). Pulsed electric field inactivation of diarrhoeagenic Bacillus cereus through irreversible electroporation. Letters in Applied Microbiology, 31(2), 110–114.

    Article  CAS  Google Scholar 

  • Satkauskas, S., & Saulis, G. (2004). Electroporation as a tool for biotechnology and medicine with specific emphasize on its application for drug and gene delivery. Review. Veterinarija ir Zootechnika, 26(48), 74.

    Google Scholar 

  • Saulis, G. (2010). Electroporation of cell membranes: the fundamental effects of pulsed electric fields in food processing. Food Engineering Reviews, 2(2), 52–73.

    Article  Google Scholar 

  • Schultheiss, C., Bluhm, H., Mayer, H.-G., Kern, M., Michelberger, T., & Witte, G. (2002). Processing of sugar beets with pulsed-electric fields. IEEE Transactions on Plasma Science, 30(4), 1547–1551.

    Article  Google Scholar 

  • Tekle, E., Astumian, R. D., & Chock, P. B. (1991). Electroporation by using bipolar oscillating electric field: an improved method for DNA transfection of NIH 3T3 cells. Proceedings of the National Academy of Sciences, 88(10), 4230–4234.

    Article  CAS  Google Scholar 

  • Vorobiev, E., & Lebovka, N. (2009). Pulsed-electric-fields-induced effects in plant tissues: fundamental aspects and perspectives of applications. Electrotechnologies for extraction from food plants and biomaterials (pp. 39–81). New York: Springer. Food Engineering Series.

    Google Scholar 

  • Vorobiev, E., Jemai, A. B., Bouzrara, H., Lebovka, N., & Bazhal, M. (2005). Pulsed electric field-assisted extraction of juice from food plants. In G. V. Barbosa-Cánovas, M. S. Tapia, & M. P. Cano (Eds.), Novel food processing technologies (pp. 105–130). New York: CRC.

    Google Scholar 

  • Weaver, J. C. (2000). Electroporation of cells and tissues. IEEE Transactions on Plasma Science, 28(1), 24–33.

    Article  CAS  Google Scholar 

  • Wouters, P. C., & Smelt, J. P. P. M. (1997). Inactivation of microorganisms with pulsed electric fields: potential for food preservation. Food Biotechnology, 11(3), 193–229.

    Article  Google Scholar 

  • Zimmermann, U., & Vienken, J. (1982). Electric field-induced cell-to-cell fusion. The Journal of Membrane Biology, 67(3), 165–182.

    Article  CAS  Google Scholar 

  • Zimmermann, U., Pilwat, G., & Riemann, F. (1974). Dielectric breakdown of cell membranes. Biophysical Journal, 14(11), 881–899.

    Article  CAS  Google Scholar 

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Acknowledgments

The research leading to these results received funding from the European Community’s Seventh Framework Program (FP7/2007-2013) under grant agreement no. 245280, also known under the acronym PRESERF.

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Correspondence to Katarzyna Dymek.

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Dymek, K., Dejmek, P. & Galindo, F.G. Influence of Pulsed Electric Field Protocols on the Reversible Permeabilization of Rucola Leaves. Food Bioprocess Technol 7, 761–773 (2014). https://doi.org/10.1007/s11947-013-1067-y

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