Postharvest Chlorophyll Degradation and Oxidative Stress

  • Naoki YamauchiEmail author


A loss of green color from chlorophyll (Chl) degradation in harvested green horticultural crops is one of the main quality problems occurring during transportation and storage. In crops such as leafy vegetables, broccoli, and lime, initial Chl a degradation was thought to lead to chlorophyllide a formation by chlorophyllase, pheophytin a formation by Mg-dechelation, or 132-hydroxychlorophyll a formation by oxidation. Oxidative degradation of Chl a could be caused by the oxidation of phenolics, which have a OH group of p-position in the benzene ring, or that of unsaturated fatty acids with senescence.

Postharvest stresses, immoderate temperature, light, and desiccation seem to induce the production of reactive oxygen species (ROS) in horticultural produce. Those environmental stresses can accelerate the advancement of senescence, including Chl degradation. In contrast, generation of an appropriate amount of ROS by stress treatments induces the activation of antioxidant enzymes and an ascorbate-glutathione cycle, which eliminates hydrogen peroxide in the cell, and in consequence, suppression of postharvest Chl degradation of green horticultural crops. Thus, oxidative stress could be seen as a double-edged sword in association with the advancement of quality deterioration throughout transportation and storage of green crops.


Antioxidant Chlorophyll degradation Quality deterioration Reactive oxygen species Senescence Stress treatments Transportation and storage 


  1. Aiamla-or S, Yamauchi N, Takino S, Shigyo M (2009) Effect of UV-A and UV-B irradiation on broccoli (Brassica oleracea L. Italica group) floret yellowing during storage. Postharvest Biol Technol 54:177–179CrossRefGoogle Scholar
  2. Aiamla-or S, Kaewsuksaeng S, Shigyo M, Yamauchi N (2010) Impact of UV-B irradiation on chlorophyll degradation and chlorophyll-degrading enzyme activities in stored broccoli (Brassica oleracea L. Italica group) florets. Food Chem 120:645–651CrossRefGoogle Scholar
  3. Aiamla-or S, Nakajima T, Shigyo M, Yamauchi N (2012) Pheophytinase activity and gene expression of chlorophyll-degrading enzyme relating to UV-B treatment in postharvest broccoli (Brassica oleracea L. Italica group) florets. Postharvest Biol Technol 63:60–66CrossRefGoogle Scholar
  4. Aiamla-or S, Shigyo M, Ito S, Yamauchi N (2014) Involvement of chloroplast peroxidase on chlorophyll degradation in postharvest broccoli florets and its control by UV-B treatment. Food Chem 165:224–231Google Scholar
  5. Akiyama Y, Takahashi C, Yamauchi N (2000) Pathway of chlorophyll degradation in yellowing radish (Raphanus sativus L.) cotyledons. J Jpn Soc Food Sci Technol 47(4):296–301CrossRefGoogle Scholar
  6. Amir-Shapira D, Goldschmidt EE, Altman A (1987) Chlorophyll catabolism in senescing plant tissues: in vivo breakdown intermediates suggest different degradative pathways for Citrus fruit and parsley leaves. Proc Natl Acad Sci USA 84(7):1901–1905PubMedCentralPubMedCrossRefGoogle Scholar
  7. Barmore CR (1975) Effect of ethylene on chlorophyllase activity and chlorophyll content in calamondin rind tissue. HortScience 10:595–596Google Scholar
  8. Blackbourn HD, Jeger MJ, John P (1990) Inhibition of degreening in the peel of bananas ripened at tropical temperatures. V. Chlorophyll bleaching activity measured in vitro. Ann Appl Biol 117(1):175–186CrossRefGoogle Scholar
  9. Büchart AM, Civello PM, Martínez GA (2010) Effect of hot air, UV-C, white light and modified atmosphere treatments on expression of chlorophyll degrading genes in postharvest broccoli (Brassica oleracea L.) florets. Sci Hortic 127(3):214–219CrossRefGoogle Scholar
  10. Büchart AM, Civello PM, Martínez GA (2011) Characterization of Mg-dechelating substance in senescent and pre-senescent Arabidopsis thaliana leaves. Biol Plant 55(1):75–82Google Scholar
  11. Buckle KA, Edwards RA (1970) Chlorophyll degradation and lipid oxidation in frozen unblanched peas. J Sci Food Agric 21(6):307–312PubMedCrossRefGoogle Scholar
  12. Chairat B, Nutthachai P, Varit S (2013) Effect of UV-C treatment on chlorophyll degradation, antioxidant enzyme activities and senescence in Chinese kale (Brassica oleracea var. alboglabra). Int Food Res J 20(2):623–628Google Scholar
  13. Chen LO, Lin C, Kelker SM, Chang Y, Shaw J (2008) Transgenic broccoli (Brassica oleracea L. var. Italica) with antisense chlorophyllase (BoCLH1) delays postharvest yellowing. Plant Sci 174(1):25–31CrossRefGoogle Scholar
  14. Cheng Y, Dong Y, Yan H, Ge W, Shen C, Guan J, Liu L, Zhang Y (2012) Effects of 1-MCP on chlorophyll degradation pathway-associated genes expression and chloroplast ultrastructure during the peel yellowing of Chinese pear fruits in storage. Food Chem 135(2):415–422PubMedCrossRefGoogle Scholar
  15. Costa ML, Civello PM, Chaves AR, Martínez GA (2002) Characterization of Mg-dechelatase activity obtained from Fragaria × ananassa fruit. Plant Physiol Biochem 40(2):111–118CrossRefGoogle Scholar
  16. Costa L, Vicente AR, Civello PM, Chaves AR, Martínez GA (2006) UV-C treatment delays postharvest senescence in broccoli florets. Postharvest Biol Technol 39(2):204–210CrossRefGoogle Scholar
  17. Dissanayake PK, Yamauchi N, Shigyo M (2008) Chlorophyll degradation and resulting catabolite formation in stored Japanese bunching onion (Allium fistulosum L.). J Sci Food Agric 88(11):1981–1986CrossRefGoogle Scholar
  18. Erkan M, Wang SY, Wang CY (2008) Effect of UV treatment on antioxidant capacity, antioxidant enzyme activity and decay in strawberry fruit. Postharvest Biol Technol 48(2):163–171CrossRefGoogle Scholar
  19. Fukasawa A, Suzuki Y, Terai H, Yamauchi N (2010) Effects of postharvest ethanol vapor treatment on activities and gene expression of chlorophyll catabolic enzymes in broccoli florets. Postharvest Biol Technol 55(2):97–102CrossRefGoogle Scholar
  20. Funamoto Y, Yamauchi N, Shigenaga T, Shigyo M (2002) Effects of heat treatment on chlorophyll degrading enzymes in stored broccoli (Brassica oleracea L.). Postharvest Biol Technol 24(2):163–170CrossRefGoogle Scholar
  21. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48(12):909–930PubMedCrossRefGoogle Scholar
  22. Gómez F, Fernández L, Gergoff G, Guiamet JJ, Chaves A, Bartoli CG (2008) Heat shock increases mitochondrial H2O2 production and extends postharvest life of spinach leaves. Postharvest Biol Technol 49(2):229–234CrossRefGoogle Scholar
  23. Gross J (1987) Pigments in fruits. Academic Press, LondonGoogle Scholar
  24. Gross J (1991) Pigments in vegetables. Chlorophylls and carotenoids. AVI, New YorkCrossRefGoogle Scholar
  25. Heaton JW, Marangoni AG (1996) Chlorophyll degradation in processed foods and senescent plant tissues. Trends Food Sci Technol 7(1):8–15CrossRefGoogle Scholar
  26. Hodges DM (2003) Postharvest oxidative stress in horticultural crops. Food Products Press, New YorkGoogle Scholar
  27. Hörtensteiner S (2013) Update on the biochemistry of chlorophyll breakdown. Plant Mol Biol 82(6):505–517PubMedCrossRefGoogle Scholar
  28. Hörtensteiner S, Kräutler B (2011) Chlorophyll breakdown in higher plants. Biochim Biophys Acta 1807(8):977–988PubMedCrossRefGoogle Scholar
  29. Hörtensteiner S, Lee DW (2007) Chlorophyll catabolism and leaf coloration. In: Gan S (ed) Senescence processes in plants. Annual Plant Reviews, vol 26. Blackwell, Oxford, pp 12–38Google Scholar
  30. Huff A (1982) Peroxidase-catalysed oxidation of chlorophyll by hydrogen peroxide. Phytochemistry 21(2):261–265CrossRefGoogle Scholar
  31. Janave MT (1997) Enzymic degradation of chlorophyll in Cavendish bananas: in vitro evidence for two independent degradative pathways. Plant Physiol Biochem 35(11):837–846Google Scholar
  32. Johnson-Flanagan AM, McLachlan G (1990) Peroxidase-mediated chlorophyll bleaching in degreening canola (Brassica napus) seeds and its inhibition by sublethal freezing. Physiol Plant 80(3):453–459CrossRefGoogle Scholar
  33. Johnson-Flanagan AM, Spencer MS (1996) Chlorophyllase and peroxidase activity during degreening of maturing canola (Brassica napus) and mustard (Brassica juncea) seed. Physiol Plant 97(2):353–359CrossRefGoogle Scholar
  34. Kato M, Shimizu S (1985) Chlorophyll metabolism in higher plants. VI. Involvement of peroxidase in chlorophyll degradation. Plant Cell Physiol 26(7):1291–1301Google Scholar
  35. Kazami D, Sato T, Nakagawa H, Ogura N (1991) Effect of pre-storage hot water dipping of broccoli heads on shelf life and quality during storage. Nippon Nõgeikagaku Kaishi 65(1):19–26CrossRefGoogle Scholar
  36. Kovtun Y, Chiu WL, Tena G, Sheen J (2000) Functional analysis of oxidative stress-activated mitogen-activated protein kinase cascade in plants. Proc Natl Acad Sci USA 97(6):2940–2945PubMedCentralPubMedCrossRefGoogle Scholar
  37. Kunieda T, Amano T, Shioi Y (2005) Search for chlorophyll degradation enzyme, Mg-dechelatase, from extracts of Chenopodium album with native and artificial substrates. Plant Sci 169(1):177–183CrossRefGoogle Scholar
  38. Kuroda M, Ozawa T, Imagawa H (1990) Changes in chloroplast peroxidase activities in relation to chlorophyll loss in barley leaf segments. Physiol Plant 80(4):555–560CrossRefGoogle Scholar
  39. Lee GC, Chepyshko H, Chen HH, Chu CC, Chou YF, Akoh CC, Shaw JF (2010) Genes and biochemical characterization of three novel chlorophyllase isozymes from Brassica oleracea. J Agric Food Chem 58(15):8651–8657PubMedCrossRefGoogle Scholar
  40. Lurie S, Mitcham EJ (2007) Physiological responses of agricultural commodities to heat treatments. In: Tang J et al (eds) Heat treatments for postharvest pest control: theory and practice. CABI, Wallingford, pp 79–104CrossRefGoogle Scholar
  41. Ma XW, Shimokawa K (1998) In vitro studies on ethylene-enhanced chlorophyll degrading peroxidase and its reaction products in banana (Musa sapientum L.) fruits. J Jpn Soc Hortic Sci 67(2):261–269CrossRefGoogle Scholar
  42. Maeda Y, Kurata H, Adachi M, Shimokawa K (1998) Chlorophyll catabolism in ethylene-treated Citrus unshiu fruits. J Jpn Soc Hortic Sci 67(4):497–502CrossRefGoogle Scholar
  43. Martínez GA, Civello PM, Chaves AR, Aňón MC (2001) Characterization of peroxidase-mediated chlorophyll bleaching in strawberry fruit. Phytochemistry 58(3):379–387PubMedCrossRefGoogle Scholar
  44. Matile P, Schellenberg M, Vicentini F (1997) Localization of chlorophyllase in the chloroplast envelope. Planta (Berl) 201(1):96–99CrossRefGoogle Scholar
  45. Matile P, Hörtensteiner S, Thomas H (1999) Chlorophyll degradation. Annu Rev Plant Physiol Plant Mol Biol 50:67–95PubMedCrossRefGoogle Scholar
  46. Maunders MJ, Brown SB, Woolhouse HW (1983) The appearance of chlorophyll derivatives in senescing tissue. Phytochemistry 22(11):2443–2446CrossRefGoogle Scholar
  47. McFeeters RF (1975) Substrate specificity of chlorophyllase. Plant Physiol 55(2):377–381PubMedCentralPubMedCrossRefGoogle Scholar
  48. McFeeters RF, Chichester CO, Whitaker JR (1971) Purification and properties of chlorophyllase from Ailanthus altissima (tree-of-heaven). Plant Physiol 47(5):609–618PubMedCentralPubMedCrossRefGoogle Scholar
  49. Mínguez-Mosquera MI, Gandul-Rojas B, Gallardo-Guerrero L (1994) Measurement of chlorophyllase activity in olive fruit (Olea europaea). J Biochem (Tokyo) 116(2):263–268Google Scholar
  50. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7(9):405–410PubMedCrossRefGoogle Scholar
  51. Mori T, Terai H, Yamauchi N, Suzuki Y (2009) Effects of postharvest ethanol vapor treatment on the ascorbate-glutathione cycle in broccoli florets. Postharvest Biol Technol 52(1):134–136CrossRefGoogle Scholar
  52. Ninomiya A, Morimoto M, Shimizu S (1987) Lipid peroxidation by the (peroxidase/H2O2/phenolic) system. Plant Cell Physiol 28(4):731–734Google Scholar
  53. Noma Y, Suzuki Y, Terai H, Yamauchi N (2004) Effects of postharvest ethanol vapor treatment on quality of sudachi (Citrus sudachi Hort. ex. Shirai) fruit. Food Preserv Sci 35(4):187–193Google Scholar
  54. Ogo Y, Umeno C, Shigyo M, Yamauchi N (2011) Control of degreening during storage by heat treatment in green sour citrus fruit. Hortic Res (Japan) 10(suppl 1):473Google Scholar
  55. Okazawa A, Tang L, Ito Y, Fukusaki E, Kobayashi A (2006) Characterization and subcellular localization of chlorophyllase from Ginkgo biloba. Z Naturforsch 61c(1/2):111–117Google Scholar
  56. Peng G, Xie XL, Jiang Q, Song S, Xu CJ (2013) Chlorophyll a/b binding protein plays a key role in natural and ethylene-induced degreening of Ponkan (Citrus reticulata Blanco). Sci Hortic 160:37–43CrossRefGoogle Scholar
  57. Schelbert S, Aubry S, Burla B, Agne B, Kessler F, Krupinska K, Hörtensteiner S (2009) Pheophytin pheophorbide hydrolase (pheophytinase) is involved in chlorophyll breakdown during leaf senescence in Arabidopsis. Plant Cell 21(3):767–785PubMedCentralPubMedCrossRefGoogle Scholar
  58. Schoch S, Rüdiger W, Lüthy B, Matile P (1984) 132-Hydroxychlorophyll a, the first product of the reaction of chlorophyll-oxidase. J Plant Physiol 115(1):85–89PubMedCrossRefGoogle Scholar
  59. Shigenaga T, Yamauchi N, Funamoto Y, Shigyo M (2005) Effects of heat treatment on an ascorbate-glutathione cycle in stored broccoli (Brassica oleracea L.) florets. Postharvest Biol Technol 38(2):152–159CrossRefGoogle Scholar
  60. Shimokawa K, Shimada S, Yaeo K (1978) Ethylene-enhanced chlorophyllase activity during degreening of Citrus unshiu Marc. Sci Hortic 8(2):129–135CrossRefGoogle Scholar
  61. Shioi Y, Tomita N, Tsuchiya T, Takamiya K (1996) Conversion of chlorophyllide to pheophorbide by Mg-dechelating substance in extracts of Chenopodium album. Plant Physiol Biochem 34Z(1):41–47Google Scholar
  62. Suzuki T, Shioi Y (2002) Re-examination of Mg-dechelation reaction in the degradation of chlorophylls using chlorophyllin a as a substrate. Photosynth Res 74(2):217–223PubMedCrossRefGoogle Scholar
  63. Suzuki T, Kunieda T, Murai F, Morioka S, Shioi Y (2005) Mg-dechelation activity in radish cotyledons with artificial and native substrates, Mg-chlorophyllin a and chlorophyllide a. Plant Physiol Biochem 43(5):459–464PubMedCrossRefGoogle Scholar
  64. Takahama U, Oniki T (1998) 3,4-Dihydroxyphenylalanine is oxidized by phenoxy radicals of hydroxycinnamic acid esters in leaves of Vicia faba L. J Plant Res 111(4):487–494CrossRefGoogle Scholar
  65. Tang L, Okazawa A, Fukusaki E, Kobayashi A (2000) Removal of magnesium by Mg-dechelatase is a major step in the chlorophyll-degrading pathway in Ginkgo biloba in the process of autumnal tints. Z Naturforsch 55c(11/12):923–926Google Scholar
  66. Tenorio MD, Villanueva MJ, Sagardoy M (2004) Changes in carotenoids and chlorophylls in fresh green asparagus (Asparagus officinalis L.) stored under modified atmosphere packaging. J Sci Food Agric 84(4):357–365CrossRefGoogle Scholar
  67. Terai H, Kanou M, Mizuno M, Tsuchida H (1999) Inhibition of yellowing and ethylene production in broccoli florets following high temperature treatment with hot air. Food Preserv Sci 25(5):221–227CrossRefGoogle Scholar
  68. Tian MS, Woolf AB, Bowen JH, Ferguson IB (1996) Changes in color and chlorophyll fluorescence of broccoli florets following hot water treatment. J Am Soc Hortic Sci 121(2):310–313Google Scholar
  69. Toivonen PMA, Hodges DM (2011) Abiotic stress in harvested fruits and vegetables. In: Shanker A (ed) Abiotic stress in plants: mechanism and adaptations. InTech, Rijeka, pp 39–58Google Scholar
  70. Tsuchiya T, Ohta H, Masuda T, Mikami B, Kita N, Shioi Y, Takamiya K (1997) Purification and characterization of two isozymes of chlorophyllase from mature leaves of Chenopodium album. Plant Cell Physiol 38(9):1026–1031CrossRefGoogle Scholar
  71. Vergara-Domínguez H, Gandul-Rojas B, Roca M (2011) Formation of oxidised chlorophyll catabolites in olives. J Food Compos Anal 24(6):851–857CrossRefGoogle Scholar
  72. Vicentini F, Iten F, Matile P (1995) Development of an assay for Mg-dechelatase of oilseed rape cotyledons, using chlorophyllin as the substrate. Physiol Plant 94(1):57–63CrossRefGoogle Scholar
  73. Yamauchi N (2013) Quality maintenance of postharvest horticultural crops by stress treatments and approach for the elucidation of its mechanism. J Jpn Soc Hortic Sci 82(1):1–10CrossRefGoogle Scholar
  74. Yamauchi N, Minamide T (1985) Chlorophyll degradation by peroxidase in parsley leaves. J Jpn Soc Hortic Sci 54(2):265–271CrossRefGoogle Scholar
  75. Yamauchi N, Watada AE (1991) Regulated chlorophyll degradation in spinach leaves during storage. J Am Soc Hortic Sci 116(1):58–62Google Scholar
  76. Yamauchi N, Watada AE (1993) Pigment changes in parsley leaves during storage in controlled or ethylene-containing atmosphere. J Food Sci 58(3):616–618CrossRefGoogle Scholar
  77. Yamauchi N, Watada AE (1998) Chlorophyll and xanthophyll changes in broccoli florets stored under elevated CO2 or ethylene-containing atmosphere. HortScience 33:114–117Google Scholar
  78. Yamauchi N, Yoshimura M, Shono Y, Kozukue N (1995) Chlorophyll degradation in mitsuba leaves during storage. J Jpn Soc Food Sci Technol 42(9):709–714CrossRefGoogle Scholar
  79. Yamauchi N, Akiyama Y, Kako S, Hashinaga F (1997) Chlorophyll degradation in wase Satsuma mandarin (Citrus unshiu Marc.) fruit with on-tree maturation and ethylene treatment. Sci Hortic 71(1-2):35–42CrossRefGoogle Scholar
  80. Yamauchi N, Funamoto Y, Shigyo M (2004) Peroxidase-mediated chlorophyll degradation in horticultural crops. Phytochem Rev 3(1-2):221–228CrossRefGoogle Scholar
  81. Yamauchi N, Aiamla-or S, Eguchi K, Shigyo M, Migita CT (2012) Effect of naringin radical formed by peroxidase on chlorophyll degradation during storage of Citrus nagato-yuzukichi fruit. J Jpn Soc Hortic Sci 81(1):117–122CrossRefGoogle Scholar
  82. Yamauchi N, Takamura K, Shigyo M, Migita CT, Masuda Y, Maekawa T (2014) Control of degreening in postharvest green sour citrus fruit by electrostatic atomized water particles. Food Chem 156:160–164PubMedCrossRefGoogle Scholar
  83. Zhang X, Zhang Z, Li J, Wu L, Guo J, Ouyang L, Xia Y, Huang X, Pang X (2011) Correlation of leaf senescence and gene expression/activities of chlorophyll degradation enzymes in harvested Chinese flowering cabbage (Brassica rapa var. parachinensis). J Plant Physiol 168(17):2081–2087PubMedCrossRefGoogle Scholar

Copyright information

© Springer Japan 2015

Authors and Affiliations

  1. 1.Faculty of AgricultureYamaguchi UniversityYamaguchiJapan

Personalised recommendations