Journal of Food Measurement and Characterization

, Volume 13, Issue 3, pp 2177–2189 | Cite as

Determination of physical, biochemical and microstructural changes in impact-bruise damaged pomegranate fruit

  • Zaharan Hussein
  • Olaniyi Amos Fawole
  • Umezuruike Linus OparaEmail author
Original Paper


This study investigated the physical, biochemical and cellular microstructural changes of ‘Wonderful’ pomegranate fruit induced by impact bruising at cold (5 °C) and ambient (20 °C) temperature. Pomegranate fruit were bruised by dropping at various drop heights, from low (20 cm), medium (40 cm) to high (60 cm) impacts onto a rigid impact surface. Physical and biochemical changes such as colour browning, peel electrolyte leakage and polyphenol oxidase (PPO) enzyme activity of bruised and non-dropped (control) fruit peels were measured. Reaction oxygen species (ROS) of fluorescent probe 2,7-dichlorodihydrofluorescein diacetate (H2DCF-DA) treated fruit peels were measured by confocal laser-scanning microscopy. Qualitative assessment of microstructures between control and bruised tissues of pomegranate fruit peels was performed using scanning electron microscope (SEM). Micrographs of SEM showed cellular microstructural differences between control and bruised fruit tissues were visible after 4 and 48 h of drop impact. Medium and high impact-bruised fruit were characterized by higher ROS than control fruit. Bruise damage to pomegranate fruit lead to increased peel electrolyte leakage (PEL) both at cold and ambient temperatures. Bruising had more effect on increasing PPO activity than did the incubation temperature and time. Activity of PPO enzyme was higher in bruised fruit than control fruit. Browning score (BS) and the total colour difference (TCD) both indicated highest values corresponding to medium and high drop impact bruising. Pearson’s correlation showed strong to moderate relationship between PEL, PPO activity, BS, TCD and ROS. Increase in fluorescent units of ROS showed a strong significant correlation (p < 0.05; r = 0.70) with BS and moderately correlated with TCD (p < 0.05; r = 0.67) and PPO activity (p < 0.05; r = 0.61). This study has confirmed that pomegranate fruit bruising induce the physical and biochemical changes in addition to underlying cellular microstructural alterations.


Bruise damage Pomegranate fruit Reaction oxygen species Polyphenol oxidase Electrolyte leakage Browning 



This work is based on the research supported wholly/in part by the National Research Foundation of South Africa (Grant No. 64813). The opinions, findings and conclusions or recommendations expressed are those of the author(s) alone, and the NRF accepts no liability whatsoever in this regard. The financial support of the Innovative Agricultural Research Initiative (iAGRI – Tanzania) and the Regional Universities Forum for Capacity Building in Agriculture (RUFORUM) through the award of postgraduate scholarship to Mr. Hussein is gratefully acknowledged.


  1. 1.
    U.L. Opara, P.B. Pathare, Bruise damage measurement and analysis of fresh horticultural produce—a review. Postharvest. Biol. Technol. 91, 9–24 (2014)Google Scholar
  2. 2.
    Y. Zhang, R. Zhang, Study on the mechanism of browning of pomegranate (Punica granatum L. cv. Ganesh) peel in different storage conditions. Agric. Sci. China 7 (1), 65–73 (2008)Google Scholar
  3. 3.
    C. Bugaud, G. Ocrisse, F. Salmon, D. Rinaldo, Bruise susceptibility of banana peel in relation to genotype and post-climacteric storage conditions. Postharvest. Biol. Technol. 87, 113–119 (2014)Google Scholar
  4. 4.
    D. Rinaldo, D. Mbéguié-A-Mbéguié, B. Fils-Lycaon, Advances on polyphenols and their metabolism in sub-tropical and tropical fruits. Trends Food Sci. Technol. 21(12), 599–606 (2010)Google Scholar
  5. 5.
    M.R. Jiménez, P. Rallo, H.F. Rapoport, M.P. Suárez, Distribution and timing of cell damage associated with olive fruit bruising and its use in analyzing susceptibility. Postharvest. Biol. Technol. 111, 117–125 (2016)Google Scholar
  6. 6.
    H.J. Lee, T. Kim, S.J. Kim, S. Park, Bruising injury of persimmon (Diospyros kaki cv. Fuyu) fruits. Sci. Hortic. 103 (2), 179–185 (2005)Google Scholar
  7. 7.
    K.A. Segovia-Bravo, M. Jarén-Galán, P. García-García, A. Garrido-Fernández, Browning reactions in olives: mechanism and polyphenols involved. Food Chem. 114(4), 1380–1385 (2009)Google Scholar
  8. 8.
    K. Mitsuhashi-Gonzalez, M.J. Pitts, J.K. Fellman, E.A. Curry, C.D. Clary, Bruising profile of fresh apples associated with tissue type and structure. Appl. Eng. Agric. 26(3), 509–517 (2010)Google Scholar
  9. 9.
    P. Ding, Y.S. Ling, Browning assessment methods and polyphenol oxidase in UV-C irradiated Berangan banana fruit. Food Res. Int. 21(4), 1667–1674 (2014)Google Scholar
  10. 10.
    K. Waliszewski, V. Pardio, L. Ovando, Control of polyphenol oxidase activity in banana slices during osmotic dehydration. Dry Technol. 25(2), 375–378 (2007)Google Scholar
  11. 11.
    G. De Martino, K. Vizovitis, R. Botondi, A. Bellincontro, F. Mencarelli, 1-MCP controls ripening induced by impact injury on apricots by affecting SOD and POX activities. Postharvest. Biol. Technol. 39(1), 38–47 (2006)Google Scholar
  12. 12.
    R. Jiménez, P. Rallo, M.P. Suárez, H.F. Rapoport, Cultivar susceptibility and anatomical evaluation of table olive fruit bruising. In J. Tous and R. Gucci (Eds.), Proceedings of the XXVIII international horticultural congress on science and horticulture (pp. 419–424). Acta Hortic. 924, ISHS (2011)Google Scholar
  13. 13.
    U. Kitthawee, S. Pathaveerat, T. Srirungruang, D. Slaughter, Mechanical bruising of young coconut. Biosyst. Eng. 109(3), 211–219 (2011)Google Scholar
  14. 14.
    R. Tabatabaekoloor, Engineering properties and bruise susceptibility of peach fruits (Prunus persica). Agric. Eng. Int. CIGR J. 15(4), 244–252 (2013)Google Scholar
  15. 15.
    W. Samim, N.H. Banks, Colour changes in bruised apple fruit tissue. N. Zeal. J. Crop Hortic. 21(4), 367–372 (1993)Google Scholar
  16. 16.
    B. Terman, U.T. Gustafsson, Brunk, Mitochondrial damage and intralysosomal degradation in cellular aging. Mol. Aspects Med. 27(5–6), 471–482 (2006)Google Scholar
  17. 17.
    J.F. Turrens, Mitochondrial formation of reactive oxygen species—topical review. J. Physiol. 552(2), 335–344 (2003)Google Scholar
  18. 18.
    F. Minibayeva, O. Kolesnikov, A. Chasov, R.P. Beckett, S. Lüthje, N. Vylegzhanina, F. Buck, M. Böttger, Wound-induced apoplastic peroxidase activities: their roles in the production and detoxification of reactive oxygen species. Plant Cell Environ. 32, 497–508 (2009)Google Scholar
  19. 19.
    R. Sabban-Amin, O. Feygenberg, E. Belausov, E. Pesis, Low oxygen and1-MCP pretreatments delay superficial scald development by reducing reactive oxygen species (ROS) accumulation in stored ‘Granny Smith’ apples. Postharvest. Biol. Technol. 62(3), 295–304 (2011)Google Scholar
  20. 20.
    Y. Lv, I.I. Tahir, M.E. Olsson, Factors affecting the content of the ursolic and oleanolic acid in apple peel: influence of cultivars, sun exposure, storage conditions, bruising and Penicillium expansum infection. J. Sci. Food Agric. 96(6), 2161–2169 (2016)Google Scholar
  21. 21.
    G. Stoilkova, S. Paunova, L. Popova, Survey of wound induced changes in cucurbita pepo leaves. Biotechnol. Equip. 23(1), 217–220 (2009)Google Scholar
  22. 22.
    T. Lu, T. Finkel, Free radicals and senescence. Exp. Cell Res. 314(9), 1918–1922 (2008)Google Scholar
  23. 23.
    J. Li, J. Yan, J. Wang, Y. Zhao, J. Cao, W. Jiang, Effects of chitosan coating on oxidative stress in bruised Yali pears (Pyrus bretschneideri Rehd.). Int. J. Food Sci. Technol. 45 (10), 2149–2154 (2010)Google Scholar
  24. 24.
    S. Ketsa, M. Koolpluksee, Some physical and biochemical characteristics of damaged pericarp of mangosteen fruit after impact. Postharvest. Biol. Technol. 2, 209–215 (1993)Google Scholar
  25. 25.
    V.M. Maia, L.C.C. Salomao, D.L. Siqueira, R. Puschman, R.V.J.G.M. Filho., P. Cecon, Physical and metabolic alterations in “Prata Ana” banana induced by mechanical damage at room temperature. Sci. Agric. 68 (1), 31–36 (2011)Google Scholar
  26. 26.
    D. Macarisin, L. Cohen, A. Eick, G. Rafael, E. Belausov, M. Wisniewski, S. Droby, Penicillium digitatum suppresses production of hydrogen peroxide in host tissue during infection of citrus fruit. Phytopathology 97(11), 1491–1500 (2007)Google Scholar
  27. 27.
    M. Sayyari, M. Babalar, S. Kalantari, M. Serrano, D. Valero, Effect of salicylic acid treatment on reducing chilling injury in stored pomegranates. Postharvest. Biol. Technol. 53(3), 152–154 (2009)Google Scholar
  28. 28.
    M.R. Safizadeh, Vacuum infiltration of polyamines reduces chilling injury and firmness loss of lemon stored at chilling temperature. Int. J. Agric. Crop Sci. 6(8), 445–451 (2013)Google Scholar
  29. 29.
    H. Meighani, M. Ghasemnezhad, D. Bakhshi, Evaluation of biochemical composition and enzymatic activities in browned arils of pomegranate fruits. Int J Hortic. Sci. Technol. 1, 53–65 (2014)Google Scholar
  30. 30.
    F. Gomez-Galindo, W. Herppich, V. Gekas, I. Sjoholm, Factors affecting quality and postharvest properties of vegetables: integration of water relations and metabolism. Crit. Rev. Food Sci. Nutr. 44(3), 139–154 (2004)Google Scholar
  31. 31.
    P.B. Pathare, U.L. Opara, F.A.J. Al-Said, Colour measurement and analysis in fresh and processed foods: a review. Food Bioprocess Tech. 6(1), 36–60 (2013)Google Scholar
  32. 32.
    O.A. Fawole, U.L. Opara, Effects of storage temperature and duration on physiological responses of pomegranate fruit. Ind. Crops Prod. 47, 300–309 (2013)Google Scholar
  33. 33.
    E. Castro-Mercado, Y. Martinez-Diaz, N. Roman-Tehandon, E. Garcia-Pineda, Biochemical analysis of reactive oxygen species production and antioxidative responses in unripe avocado (Persea americana Mill var Hass) fruits in response to wounding. Protoplasma 235(1–4), 67–76 (2009)Google Scholar
  34. 34.
    A.H. Mirdehghan, M. Rahemi, D. Martınez-Romero, F. Guillen, J.M. Valverde, P.J. Zapata, M. Serrano, D. Valero, Reduction of pomegranate chilling injury during storage after heat treatment: Role of polyamines. Postharvest. Biol. Technol. 44(1), 19–25 (2007)Google Scholar
  35. 35.
    N.M.T. Ratule, A. Osman, S.H. Ahmad, N. Saari, Development of chilling injury of Berangan banana (Musa cv. Berangan (AAA)) during storage at low temperature. J. Food Agric. Environ. 4 (1), 128–134. (2006).Google Scholar
  36. 36.
    R. Zhou, S. Su, L. Yan, Y. Li, Effect of transport vibration levels on mechanical damage and physiological responses of Huanghua pears (Pyrus pyrifolia Nakai, cv. Huanghua). Postharvest. Biol. Technol. 46 (1), 20–28 (2007)Google Scholar
  37. 37.
    M.J. Tareen, N.A. Abbasi, I.A. Hafiz, Effect of salicylic acid treatments on storage life of peach fruits cv. ‘Flordaking’. Pak J Bot. 44 (1), 119–124 (2012)Google Scholar
  38. 38.
    E. Lee, S.A. Sargent, D.J. Huber, Physiological changes in Roma-type tomato induced by mechanical stress at several ripeness stages. HortScience 42(5), 1237–1242 (2007)Google Scholar
  39. 39.
    J. Leon, E. Rojo, J.J. Sanchez-Serrano, Wound signaling in plants. J. Exp. Bot. 52(354), 1–9 (2001)Google Scholar
  40. 40.
    V.M. Martinez, J.R. Whitaker, The biochemistry and control of enzymatic browning. Trends Food Sci. Technol. 6(6), 195–200 (1995)Google Scholar
  41. 41.
    E. Valero, F. Garcia-Carmona, pH-dependent effect of sodium chloride on latent grape polyphenol oxidase. J. Agric. Food Chem. 46(7), 2447–2451 (1998)Google Scholar
  42. 42.
    K.A. Segovia-Bravo, M. Jaren-Galan, P. Garcia-Garcia, A. Garrido-Fernandez, Characterization of polyphenol oxidase from the Manzanilla cultivar (Olea europaea pomiformis) and prevention of browning reactions in bruised olive fruits. J. Agric. Food Chem. 55(16), 6515–6520 (2007)Google Scholar
  43. 43.
    F. Galli, D.D. Archbold, K.W. Pomper, Pawpaw fruit chilling injury and antioxidant protection. J. Am. Soc. Hortic. Sci. 134(4), 466–471 (2009)Google Scholar
  44. 44.
    I. Lichanporn, V. Srilaong, C. Wongs-Aree, S. Kanlayanarat, Postharvest physiology and browning of longkong (Aglaia dookkoo Griff.) fruit under ambient conditions. Postharvest. Biol. Technol. 52 (3), 294–299 (2009)Google Scholar
  45. 45.
    R.L. Shewfelt, Stress physiology: a cellular approach to quality. In: Shewfelt, R.L., Prussia, S. E. (Eds.), Postharvest Handling: A Systems Approach. (Academic Press, San Diego, 1993). pp. 257–276Google Scholar
  46. 46.
    B.G. Defilippi, B.D. Whitaker, B.M. Hess-Pierce, A.A. Kader, Development and control of scald on Wonderful pomegranate during long-term storage. Postharvest. Biol. Technol. 41(3), 234–243 (2006)Google Scholar
  47. 47.
    S.P. Tian, B.Q. Li, Y. Xu, Effects of O2 and CO2 concentrations on physiology and quality of litchi fruit in storage. Food Chem. 91(4), 659–663 (2005)Google Scholar
  48. 48.
    F. Taranto, A. Pasqualone, G. Mangini, P. Tripodi, M.M. Miazzi, S. Pavan, C. Montemurro, Polyphenol oxidases in crops: Biochemical, physiological and genetic aspects—review. Int. J. Mol. Sci. 18(2), 377 (2017)Google Scholar
  49. 49.
    H.T. Lin, Y.F. Xi, S.J. Chen, A review of enzymatic browning in fruit during storage. J. Fuzhou Univ. 9 (30), 696–703 (2013)Google Scholar
  50. 50.
    A.H. Sánchez, C. Romero, E. Ramirez, M. Brenes, Storage of mechanically harvested Manzanilla olives under controlled atmospheres. Postharvest. Biol. Technol. 81, 60–65 (2013)Google Scholar
  51. 51.
    T.B.T. Nguyen, S. Ketsa, W.G. Van Doorn, Relationship between browning and the activities of polyphenol oxidase and phenylalanine ammonia lyase in banana peel during low temperature storage. Postharvest. Biol. Technol. 30(2), 187–193 (2003)Google Scholar
  52. 52.
    Y. Jiang, Role of anthocyanins, polyphenol oxidase and phenols in lychee pericarp browning. Agric. Food Chem. 80(3), 305–310 (2000)Google Scholar
  53. 53.
    N. Ekrami-Rad, J. Khazaei, M. Khoshtaghaza, Selected mechanical properties of pomegranate peel and fruit. Int. J. Food Prop. 14(3), 570–582 (2011)Google Scholar
  54. 54.
    H. Ghaffari, H.R. Ghassemzadeh, M. Sadeghi, S. Alijani, Some physical, mechanical and chemical properties of tomato fruit related to mechanical damage and bruising models. Biol. Forum 7, 712–718 (2015)Google Scholar
  55. 55.
    V. Van Linden, N. Scheerlinck, M. Desmet, J. De Baerdemaeker, Factors that affect tomato bruise development because of mechanical impact. Postharvest. Biol. Technol. 42 (3), 260–270 (2006)Google Scholar
  56. 56.
    N.K. Kim, Y.C. Hung, Microscopic measurement of apple bruise. Food Struct. 9(2), 97–104 (1990)Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Zaharan Hussein
    • 1
  • Olaniyi Amos Fawole
    • 2
  • Umezuruike Linus Opara
    • 1
    • 2
    Email author
  1. 1.Postharvest Technology Research Laboratory, South African Research Chair in Postharvest Technology, Department of Food Science, Faculty of AgriSciencesStellenbosch UniversityStellenboschSouth Africa
  2. 2.Postharvest Technology Research Laboratory, South African Research Chair in Postharvest Technology, Department of Horticultural Science, Faculty of AgriSciencesStellenbosch UniversityStellenboschSouth Africa

Personalised recommendations