Influence of Pulsed Electric Field Protocols on the Reversible Permeabilization of Rucola Leaves
- 447 Downloads
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.
KeywordsPlant tissue Electroporation Electrical resistance Survival
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.
- Bouzrara, H., & Vorobiev, E. (2000). Beet juice extraction by pressing and pulsed electric fields. International Sugar Journal, 102(1216), 194–200.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.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.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.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
- 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.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).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
- 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