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

Abstract

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.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  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 40:1131–1134

    CAS  Article  Google Scholar 

  2. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126

    CAS  Article  PubMed  Google 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 54:8335–8339

    CAS  Article  PubMed  Google Scholar 

  4. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399

    CAS  Article  PubMed  Google 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 11:551–563

    CAS  Article  Google Scholar 

  6. Asada K (1984) Chloroplasts-formation of active oxygen species. Methods Enzymol 105:422–429

    CAS  Article  Google Scholar 

  7. Asada K (1999) The water–water cycle in chloroplasts: scavenging of active oxygens and dissipation of excess photons. Annu Rev Plant Biol 50:601–639

    CAS  Article  Google 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 6:1–24

    CAS  Article  Google 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 112:1519–1531

    CAS  Article  PubMed  Google 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 03:639–650

    CAS  Article  Google 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 72:248–254

    CAS  Article  PubMed  Google Scholar 

  12. Brennan T, Frenkel C (1977) Involvement of hydrogen peroxide in the regulation of senescence in pear. Plant Physiol 59:411–416

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  13. Cocetta G, Karppinen K, Suokas M et al (2012) Ascorbic acid metabolism during bilberry (Vaccinium myrtillus L.) fruit development. J Plant Physiol 169:1059–1065

    CAS  Article  PubMed  Google 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 15:316–328

    Article  PubMed  Google 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 32:93–101

    CAS  Article  Google Scholar 

  16. Echeverria E, Burns JK (1989) Vacuolar acid hydrolysis as a physiological mechanism for sucrose breakdown. Plant Physiol 90:530–533

    PubMed Central  CAS  Article  PubMed  Google 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 54:4397–4406

    CAS  Article  PubMed  Google 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 150:37–46

    CAS  Article  Google Scholar 

  19. Foyer CH, Lelandais M, Kunert KJ (1994) Photooxidative stress in plants. Physiol Plant 92:696–717

    CAS  Article  Google 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 100:241–254

    CAS  Article  Google 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 19:43–55

    CAS  Article  Google Scholar 

  22. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930

    CAS  Article  PubMed  Google Scholar 

  23. Gillespie KM, Ainsworth EA (2007) Measurement of reduced, oxidized and total ascorbate content in plants. Nat Protoc 2:871–874

    CAS  Article  PubMed  Google Scholar 

  24. Gulsen O, Roose ML (2001) Chloroplast and nuclear genome analysis of the parentage of lemons. J Am Soc Hortic Sci 126:210–215

    CAS  Google Scholar 

  25. Haleng J, Pincemail J, Defraigne J-O et al (2007) Le stress oxydant. Rev Med Liege 62:628–638

    CAS  PubMed  Google 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 405–448

    Google 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 207:604–611

    CAS  Article  Google Scholar 

  28. Hodges DM, Lester GE, Munro KD, Toivonen PMA (2004) Oxidative stress: importance for postharvest quality. HortScience 39:924–929

    CAS  Google 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 113:166–172

    CAS  Article  Google Scholar 

  30. Jiménez A, Hernández JA, Barceló AR et al (1998) Mitochondrial and peroxisomal ascorbate peroxidase of pea leaves. Physiol Plant 104:687–692

    Article  Google 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 219:243–252

    CAS  Article  PubMed  Google Scholar 

  32. López AP, Gochicoa MTN, Franco AR (2010) Activities of antioxidant enzymes during strawberry fruit development and ripening. Biol Plant 54:349–352

    Article  Google Scholar 

  33. Lurie S (2003) Antioxidants. In: Hodges DM (ed) Postharvest oxidative stress in horticultural crops. Haworth Press, Inc., New York, pp 131–150

  34. Medlicott AP, Reynolds SB, New SW, Thompson AK (1988) Harvest maturity effects on mango fruit ripening. Trop Agric 65:153–157

    Google Scholar 

  35. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410

    CAS  Article  PubMed  Google Scholar 

  36. Mondal K, Sharma NS, Malhotra SP et al (2004) Antioxidant systems in ripening tomato fruits. Biol Plant 48:49–53

    CAS  Article  Google Scholar 

  37. Muller ML, Taiz L (2002) Regulation of the lemon fruit V-ATPase by variable stoichiometry and organic acids. J Membr Biol 185:209–220

    CAS  Article  PubMed  Google 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 100:1155–1166

    CAS  Article  Google 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 50:1093–1106

    CAS  Article  PubMed  Google Scholar 

  40. Oberley LW, Spitz DR (1984) Assay of superoxide dismutase activity in tumor tissue. Methods Enzymol 105:457–464

    CAS  Article  PubMed  Google 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 30:872–880

    CAS  Article  Google 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 149:175–187

    CAS  Article  PubMed  Google 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 1:3159–3165

    CAS  Article  Google Scholar 

  44. Richardson AC, Marsh KB, Macrae EA (1997) Temperature effects on satsuma mandarin fruit development. J Hortic Sci 72:919–929

    Google Scholar 

  45. Sala JM (1998) Involvement of oxidative stress in chilling injury in cold-stored mandarin fruits. Postharvest Biol Techn 13:255–261

    CAS  Article  Google 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 47:2410–2414

    CAS  Article  PubMed  Google 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 27:71–83

    CAS  Article  Google 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 358:149–152

    CAS  Article  PubMed  Google Scholar 

  49. Tompkins D, Toffaletti J (1982) Enzymic determination of citrate in serum and urine, with use of the Worthington “ultrafree” device. Clin Chem 28:192–195

    CAS  PubMed  Google Scholar 

  50. Tzur A, Goren R, Zehavi U (1992) Carbohydrate metabolism in developing citrus fruits. Proc Int Soc Citric 1:405–411

    Google 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 121:306–312

    CAS  Article  Google 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 49:1612–1619

    CAS  Article  PubMed  Google 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 49:113–118

    Article  Google Scholar 

Download references

Acknowledgments

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

Author information

Affiliations

Authors

Corresponding author

Correspondence to Jérémie Santini.

Additional information

Communicated by P. K. Nagar.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 227 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Oustric, J., Antoine, S., Giannettini, J. et al. Biochemical characterization of the primary metabolism and antioxidant defense systems of acidic and acidless citrus genotypes during the major stages of fruit growth. Acta Physiol Plant 37, 228 (2015). https://doi.org/10.1007/s11738-015-1982-9

Download citation

Keywords

  • Citrus
  • Sugars
  • Organic acids
  • Antioxidant system
  • Oxidative status