Acta Physiologiae Plantarum

, 37:228 | Cite as

Biochemical characterization of the primary metabolism and antioxidant defense systems of acidic and acidless citrus genotypes during the major stages of fruit growth

  • Julie Oustric
  • Sandrine Antoine
  • Jean Giannettini
  • Yves Gibon
  • François Luro
  • Liliane Berti
  • Jérémie SantiniEmail author
Original Article


Fruits are consumed not just for their taste but also for their nutritional value. The major primary metabolites in fruit are sugars and acids, whose contents change during fruit growth and determine ultimate fruit quality. Fruits are also a source of antioxidant metabolites, which are important to human health due to their role in reducing risk of cancer and cardiovascular diseases. Antioxidants are equally important in the plant as they help fight against oxidative stress. Here, we investigated the consequences of changes in the primary metabolism in acidic and acidless citrus genotypes during the major stages of fruit growth on the expression of antioxidant enzymes and the markers of cellular oxidation (hydrogen peroxide, malondialdehyde) in acidless (Iaffaoui orange and sweet lemon) and acidic (Salustiana orange and Villafranca lemon) citrus fruits. Glucose and fructose were the major sugars in the acidless lemon. Sucrose was the major sugar in the acidic lemon. Oranges shared a balance of glucose, fructose, and sucrose. Malic and citric acid concentrations were higher in acidic lemons than acidless fruits. Acidic genotypes had higher hydrogen peroxide concentrations than acidless genotypes, whereas MDA concentrations were higher in oranges than in lemons. Specific activities of ascorbate peroxidase, catalase, superoxide dismutase, and dehydroascorbate reductase were on the whole higher in acidic than acidless fruits. Principal component analysis revealed between-genotype divergence in antioxidant system, giving three groups: acidic lemons, acidless lemons, and oranges.


Citrus Sugars Organic acids Antioxidant system Oxidative status 



The authors thank the Collectivité Territoriale de Corse (CTC) for providing financial support for this study.

Supplementary material

11738_2015_1982_MOESM1_ESM.docx (228 kb)
Supplementary material 1 (DOCX 227 kb)


  1. Ackermann J, Fischer M, Amado R (1992) Changes in sugars, acids, and amino acids during ripening and storage of apples (cv. Glockenapfel). J Agric Food Chem 40:1131–1134CrossRefGoogle Scholar
  2. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefPubMedGoogle Scholar
  3. Albertini M-V, Carcouet E, Pailly O et al (2006) Changes in organic acids and sugars during early stages of development of acidic and acidless citrus fruit. J Agric Food Chem 54:8335–8339CrossRefPubMedGoogle Scholar
  4. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399CrossRefPubMedGoogle Scholar
  5. Aprile A, Federici C, Close TJ et al (2011) Expression of the H+-ATPase AHA10 proton pump is associated with citric acid accumulation in lemon juice sac cells. Funct Integr Genom 11:551–563CrossRefGoogle Scholar
  6. Asada K (1984) Chloroplasts-formation of active oxygen species. Methods Enzymol 105:422–429CrossRefGoogle Scholar
  7. Asada K (1999) The water–water cycle in chloroplasts: scavenging of active oxygens and dissipation of excess photons. Annu Rev Plant Biol 50:601–639CrossRefGoogle Scholar
  8. Bain JM (1958) Morphological, anatomical, and physiological changes in the developing fruit of the Valencia orange, Citrus sinensis (L) Osbeck. Aust J Bot 6:1–24CrossRefGoogle Scholar
  9. Barkley NA, Roose ML, Krueger RR, Federici CT (2006) Assessing genetic diversity and population structure in a citrus germplasm collection utilizing simple sequence repeat markers (SSRs). Theor Appl Genet 112:1519–1531CrossRefPubMedGoogle Scholar
  10. Bermejo A (2012) Analysis of nutritional constituents in twenty citrus cultivars from the Mediterranean area at different stages of ripening. Food Nut Sci 03:639–650CrossRefGoogle Scholar
  11. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  12. Brennan T, Frenkel C (1977) Involvement of hydrogen peroxide in the regulation of senescence in pear. Plant Physiol 59:411–416PubMedCentralCrossRefPubMedGoogle Scholar
  13. Cocetta G, Karppinen K, Suokas M et al (2012) Ascorbic acid metabolism during bilberry (Vaccinium myrtillus L.) fruit development. J Plant Physiol 169:1059–1065CrossRefPubMedGoogle Scholar
  14. Del Rio D, Stewart AJ, Pellegrini N (2005) A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr Metab Cardiovasc Dis 15:316–328CrossRefPubMedGoogle Scholar
  15. Dhindsa RS, Plumb-Dhindsa P, Thorpe TA (1981) Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. J Exp Bot 32:93–101CrossRefGoogle Scholar
  16. Echeverria E, Burns JK (1989) Vacuolar acid hydrolysis as a physiological mechanism for sucrose breakdown. Plant Physiol 90:530–533PubMedCentralCrossRefPubMedGoogle Scholar
  17. Fanciullino A-L, Dhuique-Mayer C, Luro F et al (2006) Carotenoid diversity in cultivated citrus is highly influenced by genetic factors. J Agric Food Chem 54:4397–4406CrossRefPubMedGoogle Scholar
  18. Fawole OA, Opara UL (2013) Changes in physical properties, chemical and elemental composition and antioxidant capacity of pomegranate (cv. Ruby) fruit at five maturity stages. Sci Hort 150:37–46CrossRefGoogle Scholar
  19. Foyer CH, Lelandais M, Kunert KJ (1994) Photooxidative stress in plants. Physiol Plant 92:696–717CrossRefGoogle Scholar
  20. Foyer CH, Lopez-Delgado H, Dat JF, Scott IM (1997) Hydrogen peroxide-and glutathione-associated mechanisms of acclimatory stress tolerance and signalling. Physiol Plant 100:241–254CrossRefGoogle Scholar
  21. Geigenberger P, Lerchi J, Stitt M, Sonnewald U (1996) Phloem-specific expression of pyrophosphatase inhibits long distance transport of carbohydrates and amino acids in tobacco plants. Plant Cell Environ 19:43–55CrossRefGoogle Scholar
  22. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930CrossRefPubMedGoogle Scholar
  23. Gillespie KM, Ainsworth EA (2007) Measurement of reduced, oxidized and total ascorbate content in plants. Nat Protoc 2:871–874CrossRefPubMedGoogle Scholar
  24. Gulsen O, Roose ML (2001) Chloroplast and nuclear genome analysis of the parentage of lemons. J Am Soc Hortic Sci 126:210–215Google Scholar
  25. Haleng J, Pincemail J, Defraigne J-O et al (2007) Le stress oxydant. Rev Med Liege 62:628–638PubMedGoogle Scholar
  26. Hamilton GA (1974) Chemical models and mechanism for oxygenases. In: Hayaishi O (ed) Molecular mechanisms of oxygen activation. Academic Press, New York, pp 405–448Google Scholar
  27. Hodges DM, DeLong JM, Forney CF, Prange RK (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207:604–611CrossRefGoogle Scholar
  28. Hodges DM, Lester GE, Munro KD, Toivonen PMA (2004) Oxidative stress: importance for postharvest quality. HortScience 39:924–929Google Scholar
  29. Huang R, Xia R, Hu L et al (2007) Antioxidant activity and oxygen-scavenging system in orange pulp during fruit ripening and maturation. Sci Hort 113:166–172CrossRefGoogle Scholar
  30. Jiménez A, Hernández JA, Barceló AR et al (1998) Mitochondrial and peroxisomal ascorbate peroxidase of pea leaves. Physiol Plant 104:687–692CrossRefGoogle Scholar
  31. Katz E, Lagunes PM, Riov J et al (2004) Molecular and physiological evidence suggests the existence of a system II-like pathway of ethylene production in non-climacteric Citrus fruit. Planta 219:243–252CrossRefPubMedGoogle Scholar
  32. López AP, Gochicoa MTN, Franco AR (2010) Activities of antioxidant enzymes during strawberry fruit development and ripening. Biol Plant 54:349–352CrossRefGoogle Scholar
  33. Lurie S (2003) Antioxidants. In: Hodges DM (ed) Postharvest oxidative stress in horticultural crops. Haworth Press, Inc., New York, pp 131–150Google Scholar
  34. Medlicott AP, Reynolds SB, New SW, Thompson AK (1988) Harvest maturity effects on mango fruit ripening. Trop Agric 65:153–157Google Scholar
  35. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410CrossRefPubMedGoogle Scholar
  36. Mondal K, Sharma NS, Malhotra SP et al (2004) Antioxidant systems in ripening tomato fruits. Biol Plant 48:49–53CrossRefGoogle Scholar
  37. Muller ML, Taiz L (2002) Regulation of the lemon fruit V-ATPase by variable stoichiometry and organic acids. J Membr Biol 185:209–220CrossRefPubMedGoogle Scholar
  38. Nicolosi E, Deng ZN, Gentile A et al (2000) Citrus phylogeny and genetic origin of important species as investigated by molecular markers. Theor Appl Genet 100:1155–1166CrossRefGoogle Scholar
  39. Nunes-Nesi A, Carrari F, Gibon Y et al (2007) Deficiency of mitochondrial fumarase activity in tomato plants impairs photosynthesis via an effect on stomatal function. Plant J 50:1093–1106CrossRefPubMedGoogle Scholar
  40. Oberley LW, Spitz DR (1984) Assay of superoxide dismutase activity in tumor tissue. Methods Enzymol 105:457–464CrossRefPubMedGoogle Scholar
  41. Peroni LA, Ferreira RR, Figueira A et al (2007) Expression profile of oxidative and antioxidative stress enzymes based on ESTs approach of citrus. Genet Mol Biol 30:872–880CrossRefGoogle Scholar
  42. Poiroux-Gonord F, Santini J, Fanciullino A-L et al (2013) Metabolism in orange fruits is driven by photooxidative stress in the leaves. Physiol Plant 149:175–187CrossRefPubMedGoogle Scholar
  43. Rahman I, Kode A, Biswas SK (2006) Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat Prot 1:3159–3165CrossRefGoogle Scholar
  44. Richardson AC, Marsh KB, Macrae EA (1997) Temperature effects on satsuma mandarin fruit development. J Hortic Sci 72:919–929Google Scholar
  45. Sala JM (1998) Involvement of oxidative stress in chilling injury in cold-stored mandarin fruits. Postharvest Biol Techn 13:255–261CrossRefGoogle Scholar
  46. Sala JM, Lafuente MT (1999) Catalase in the heat-induced chilling tolerance of cold-stored hybrid Fortune mandarin fruits. J Agric Food Chem 47:2410–2414CrossRefPubMedGoogle Scholar
  47. Santini J, Giannettini J, Pailly O et al (2013) Comparison of photosynthesis and antioxidant performance of several Citrus and Fortunella species (Rutaceae) under natural chilling stress. Trees 27:71–83CrossRefGoogle Scholar
  48. Schantz M-L, Schreiber H, Guillemaut P, Schantz R (1995) Changes in ascorbate peroxidase activities during fruit ripening in Capsicum annuum. FEBS Lett 358:149–152CrossRefPubMedGoogle Scholar
  49. Tompkins D, Toffaletti J (1982) Enzymic determination of citrate in serum and urine, with use of the Worthington “ultrafree” device. Clin Chem 28:192–195PubMedGoogle Scholar
  50. Tzur A, Goren R, Zehavi U (1992) Carbohydrate metabolism in developing citrus fruits. Proc Int Soc Citric 1:405–411Google Scholar
  51. Uzun A, Yesiloglu T, Tuzcu O, Gulsen O (2009) Genetic diversity and relationships within Citrus and related genera based on sequence related amplified polymorphism markers (SRAPs). Sci Hort 121:306–312CrossRefGoogle Scholar
  52. Wang SY, Jiao H (2001) Changes in oxygen-scavenging systems and membrane lipid peroxidation during maturation and ripening in blackberry. J Agric Food Chem 49:1612–1619CrossRefPubMedGoogle Scholar
  53. Zhou B, Wang J, Guo Z et al (2006) A simple colorimetric method for determination of hydrogen peroxide in plant tissues. Plant Growth Regul 49:113–118CrossRefGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2015

Authors and Affiliations

  • Julie Oustric
    • 1
  • Sandrine Antoine
    • 1
    • 2
  • Jean Giannettini
    • 1
  • Yves Gibon
    • 3
    • 4
  • François Luro
    • 2
  • Liliane Berti
    • 1
  • Jérémie Santini
    • 1
    Email author
  1. 1.CNRS, UMR 6134 SPE, Laboratoire Biochimie and Biologie Moléculaire du VégétalCorteFrance
  2. 2.UMR AGAP Corse, Station INRASan GiulianoFrance
  3. 3.Institut National de la Recherche Agronomique and Université de Bordeaux, Unité Mixte de Recherche 1332, Biologie du Fruit et PathologieVillenave-d’OrnonFrance
  4. 4.Metabolomics Platform–Functional Genomics Centre Bordeaux, INRA BordeauxVillenave-d’OrnonFrance

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