Accumulation of Naringin and Limonin and Genes Expression of their Corresponding Enzymes during Fruit Maturation of Three Citrus Genotypes

Document Type : Research Paper

Authors

1 Department of Horticulture, Faculty of Agriculture, University of Tabriz, Tabriz, Iran

2 Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz, Iran

3 Citrus and Subtropical Fruit Research Center, Ramsar, Iran

Abstract

Naringin and limonin are important antioxidant compounds in Citrus species. Due to the high biological effects of these compounds, in the present study, the variation of naringin and limonin content during fruit development was investigated at two fruit tissues of pulp and albedo in three Citrus genotypes (grapefruit, orange and mandarin). Also, the gene expression of involved enzymes in naringin and limonin metabolism was assayed. In all studied Citrus genotypes, the naringin content in albedo tissue was higher than the pulp and grapefruits showed the highest naringin content (940.26 mg per 100 g DW) followed by orange (791.17 mg per 100 g DW) and mandarin (602.10 mg per 100 g DW). The naringin content of studied genotypes was decreased during the fruit development and the highest decrement was observed at ripening stages. The decreasing of naringin content was attributed to the changes in genes expression of main involved enzymes in naringin biosynthesis pathways including chalcone-flavanone isomerase and naringenin glycoside biosynthesis (1.2 RhaT) during studied harvesting times. In both tissues of mandarin and orange fruits, the limonin content was increased up to 80 days after full bloom then decreased. In grapefruit, due to the late-ripening of the fruit, the peak of limonin increment was occurred later, which can be attributed to the long growth process of grapefruits compared to orange and mandarin. The reduction of limonin content with fruit ripening was in accordance with the increased gene expression of limonoid UDP-glucosyl transferase that converts limonin to tasteless compounds.

Keywords


  1. Iglesias I., Echeverría G. Differential effect of cultivar and harvest date on nectarine colour, quality and consumer acceptance. Sci Hortic. 2009;120:41-50.
  2. Terol J., Soler G., Talon M., Cercos M. The aconitate hydratase family from Citrus. BMC Plant Biol. 2010;10:1-12.
  3. Frydman A., Liberman R., Huhman D.V., Carmeli Weissberg M., Sapir Mir M., Ophir R., Sumner L., Eyal Y. The molecular and enzymatic basis of bitter/nonbitter flavor of citrus fruit: Evolution of branch forming rhamnosyltransferases under domestication. Plant J. 2013;73:166–178.
  4. Wang F., Wang M., Liu X., Xu Y., Zhu S., Shen W., Zhao X. Identification of putative genes involved in limonoids biosynthesis in citrus by comparative transcriptomic analysis. Front Plant Sci. 2017;8:782.
  5. Ververidis F., Trantas E., Douglas C., Vollmer G., Kretzschmar G., Panopoulos N. Biotechnology of flavonoids and other phenylpropanoid derived natural products. Part I: Chemical diversity, impacts on plant biology and human health. Biotechnol J Healthc Nutr Technol. 2007;2:1214–1234.
  6. Wang S., Tu H., Wan J., Chen W., Liu X., Luo J., Xu J., Zhang H. Spatio-temporal distribution and natural variation of metabolites in citrus fruits. Food Chem. 2016;199:8–17.
  7. Bai J., Li S., Wu G., Ma L., Li C., Liu H., Zhang P. Naringin inhibits lipopolysaccharide-induced activation of microglia cells. Cell Mol Biol. 2019;65:38–42.
  8. Farooqi A.A., Tahir F., Fakhar M., Butt G., Colombo Pimentel T., Wu N., Yulaevna I.M., Attar R. Antimetastatic effects of Citrus-derived bioactive ingredients: Mechanistic insights. Cell Mol Biol. 2021;67:178–186.
  9. Dea S., Plotto A., Manthey J.A., Raithore S., Irey M., Baldwin E. Interactions and thresholds of limonin and nomilin in bitterness perception in orange juice and other matrices. J Sens Stud. 2013;28:311–323.
  10. Zaare-Nahandi F., Hosseinkhani S., Zamani Z., Asadi-Abkenar A., Omidbaigi R. Delay expression of limonoid UDP-glucosyltransferase makes delayed bitterness in citrus. Biochem Biophys Res Commun. 2008;371:59–62.
  11. Zandalinas S.I., Sales C., Beltrán J., Gómez-Cadenas A., Arbona V. Activation of secondary metabolism in citrus plants is associated to sensitivity to combined drought and high temperatures. Front Plant Sci. 2017;7:1954.
  12. Barreca D., Bellocco E., Caristi C., Leuzzi U.G.O., Gattuso G. Flavonoid composition and antioxidant activity of juices from chinotto (Citrus × myrtifolia Raf.) fruits at different ripening stages. J Agric Food Chem. 2010;58:3031–3036.
  13. Chaudhary P.R., Bang H., Jayaprakasha G.K., Patil B.S. Variation in key flavonoid biosynthetic enzymes and phytochemicals in ‘Rio Red’grapefruit (Citrus paradisi Macf.) during fruit development. J Agric Food Chem. 2016;64:9022–9032.
  14. Asikin Y., Taira I., Inafuku S., Sumi H., Sawamura M., Takara K., Wada K. Volatile aroma components and antioxidant activities of the flavedo peel extract of unripe Shiikuwasha (Citrus depressa Hayata). J Food Sci. 2012;77:C469–C475.
  15. Liu C., Yan F., Gao H., He M., Wang Z., Cheng Y., Deng X., Xu J. Features of citrus terpenoid production as revealed by carotenoid, limonoid and aroma profiles of two pummelos (Citrus maxima) with different flesh color. J Sci Food Agric. 2015;95:111–119.
  16. Chang S.Q., Azrina A. Antioxidant content and activity in different parts of pomelo [Citrus grandis (L.) Osbeck] by-products. III International Conference on Agricultural and Food Engineering. 2016;27–34.
  17. Arbona V., Iglesias D.J., Gómez-Cadenas A. Non-targeted metabolite profiling of citrus juices as a tool for variety discrimination and metabolite flow analysis. BMC Plant Biol. 2015;15:1–16.
  18. Fallah F., Nokhasi F., Ghaheri M., Kahrizi D., Agha A.B., Ghorbani T., Kazemi E., Ansarypour Z. Effect of salinity on gene expression, morphological and biochemical characteristics of Stevia rebaudiana Bertoni under in vitro conditions. Cell Mol Biol. 2017;15:63:102-106.
  19. Kahrizi D., Ghari S.M., Ghaheri M., Fallah F., Ghorbani T., Agha A.B., Kazemi E., Ansarypour Z. Effect of KH2PO4 on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions. Cellular and Molecular Biology. 2017;15:63:107-111.
  20. Sun C., Chen K., Chen Y., Chen Q. Contents and antioxidant capacity of limonin and nomilin in different tissues of citrus fruit of four cultivars during fruit growth and maturation. Food Chem. 2005;93:599–605.
  21. Bermejo A., Llosá M.J., Cano A. Analysis of bioactive compounds in seven citrus cultivars. Food Sci Technol Int Sage Publications Sage UK: London, England. 2011;17:55–62.
  22. Bilal H., Akram W., Hassan S.A., Sahar S., Munir Iqbal M. Determination of limonin and nomilin contents in different citrus cultivars using high performance liquid chromatography. Pak J Sci Ind Res. 2013;56:36-40.
  23. Huang S., Liu X., Xiong B., Qiu X., Sun G., Wang X., Zhang X., Dong Z., Wang Z. Variation in limonin and nomilin content in citrus fruits of eight varieties determined by modified HPLC. Food Sci Biotechnol. 2019;28:641–647.
  24. Raithore S., Dea S., McCollum G., Manthey J.A., Bai J., Leclair C., Hijaz F., Narciso A.J., Baldwin E.A., Plotto A. Development of delayed bitterness and effect of harvest date in stored juice from two complex citrus hybrids. J Sci Food Agric. 2015;96:2.
  25. Wang Y., Li J., Xia R. Expression of chalcone synthase and chalcone isomerase genes and accumulation of corresponding flavonoids during fruit maturation of Guoqing No. 4 satsuma mandarin (Citrus unshiu Marcow). Sci Hortic. 2010;125:110-116.
  26. Lewinsohn E., Britsch L., Mazur Y., Gresse J. Flavanone glycoside biosynthesis in citrus. Plant Physiol. 1989;91:1323-1328.
  27. Ahmed W., Ahmad S., Malik A.U., Ahmed R. Harvesting times modulates the human health promotoing substances in grapefruit. Pak J Agric Sci. 2015;52:393-401.
  28. Moriguchi T., Kita M., Tomono Y., Endo-Inagaki T., Omura M. Gene expression in flavonoid biosynthesis: correlation with flavonoid accumulation in developing citrus fruit. Plant Physiol. 2001;111:66-74.
  29. Pichaiyongvongdee S., Haruenkit R. Comparative studies of limonin and naringin distribution in different parts of Pummelo [Citrus grandis (L.) Osbeck] cultivars grown in Thailand. Nat Sci. 2009;43:28-36.
  30. Daniel J.J., Owens D.K., McIntosh C.A. Secondary product glucosyltransferase and putative glucosyltransferase expression during Citrus paradisi (c.v. Duncan) growth and development. Phytochem. 2001;72:1732-1738.
  31. Bai J., Baldwin E.A., McCollum G., Plotto A., Manthey J.A., Widmer W.W., Luzio G., Cameron R. Changes in volatile and non-volatile flavor chemicals of “Valencia” orange juice over the harvest seasons. Foods. 2016;5:4.