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Establishment of Punica granatum L. peel cell culture to produce bioactive compounds

  • Rubinovich LiorEmail author
  • Segev Barak
  • Habashi Rida
  • Con Pazit
  • Amir Rachel
Original Article
  • 91 Downloads

Abstract

The pomegranate (Punica granatum L.) fruit harbors remarkable health-beneficial properties. Most of its healthy secondary metabolites are located in their peels. Therefore, pomegranate peels (PPs) can be used to produce high-value healthy compounds. PP cell cultures may also be an attractive alternative source since it can be established throughout the year regardless of seasonal effects. The aim of this study was to establish a novel PP cell culture for future research and biotechnological applications. We first established procedures to sterilize and control oxidative browning of the peel explants. We found that surface sterilizing with 3% sodium hypochlorite for 15 min, frequent sub-culturing of the explants on media containing polyvinylpyrrolidone plus silver nitrate was the optimal treatment for oxidative browning control and callus initiation. We also found that 6-benzylaminopurine (BAP) induced callus initiation, whereas 3-indoleacetic acid (IAA) repressed it. The callus daily growth rate was maintained at 1% regardless of the various growth media examined. Moreover, our results show that although there was a minor effect of the growth media on polyphenol accumulation and antioxidant activity, the exposure to light and high sucrose levels led to higher levels (up to 205 times than the control) of secondary metabolites such as anthocyanins, ellagic acid, punicalagin and gallic acid. In summary, this work laid the foundations for further research that will be needed to study and commercialize PP cell cultures.

Key message

In this present study, we addresses the establishment of pomegranate peel cell cultures for future research and biotechnological applications, such as production of health-beneficial secondary metabolites.

Keywords

Cell culture Pomegranate peel Punica granatum Secondary metabolites 

Notes

Acknowledgements

The authors wish to thank the Israel Ministry of Economics for funding under the Kamin Fund (No. 53856) and BARD (US-Israel Binational Agricultural Research and Development Fund 364) for funding under Project No. IS-4822-15 R.

Authors contribution

L.R. was the head of this project and was the main writer of this manuscript. B.S., R.H. and P.C. were the lab technicians which carried out most of the tissue culture work. R.A. is the PI and contributed to the planning of the experiments and data analysis.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11240_2019_1609_MOESM1_ESM.jpg (82 kb)
Supplementary material 1 (JPEG 81 kb). Supplemental Fig. A1 Antioxidant capacity of callus and pomegranate fruit skin expressed as Trolox equivalent. Values are means ± SE of four biological replicates (four Petri dishes/fruits, each dish containing nine explants). The experiment was repeated at least three times, with similar results. Significant differences (Tukey-HSD, p < 0.05) are indicated by different letters above the columns
11240_2019_1609_MOESM2_ESM.jpg (40 kb)
Supplementary material 2 (JPEG 39 kb). Supplemental Fig. A2 Anthocyanin localized in the vacuole of pomegranate peel tissue culture cells elicited with sucrose at 5%

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Northern R&DMIGAL – Galilee Research InstituteKiryat ShmonaIsrael
  2. 2.MIGAL – Galilee Research InstituteKiryat ShmonaIsrael
  3. 3.Tel Hai CollegeKiryat ShmonaIsrael

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