Skip to main content
Log in

Cloning and characterization of the cDNA for lycopene β-cyclase from tomato reveals decrease in its expression during fruit ripening

  • Research Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

The cDNA which encodes lycopene cyclase, CrtL, was cloned from tomato (Lycopersicon esculentum cv. VF36) and tobacco (Nicotiana tabacum cv. Samsun NN) and functionally expressed in Escherichia coli. This enzyme converts lycopene to β-carotene by catalyzing the formation of two β-rings at each end of the linear carotene. The enzyme interacts with half of the carotenoid molecule and requires a double bond at the C-7,8 (or C-7,8′) position. Inhibition in E. coli indicated that lycopene cyclase is the target site for the inhibitor MPTA, 2-(4-methylphenoxy)tri-ethylamine hydrochloride. The primary structure of lycopene cyclase in higher plants is significantly conserved with the enzyme from cyanobacteria but different from that of from the non-photosynthetic bacteria Erwinia. mRNA of CrtL and Pds, which encodes phytoene desaturase, was measured in leaves, flowers and ripening fruits of tomato. In contrast to genes which encode enzymes of early steps in the carotenoid biosynthesis pathway, whose transcription increases during the ‘breaker’ stage of fruit ripening, the level of CrtL mRNA decreases at this stage. Hence, the accumulation of lycopene in tomato fruits is apparently due to a down-regulation of the lycopene cyclase gene that occurs at the breaker stage of fruit development. This conclusion supports the hypothesis that transcriptional regulation of gene expression is a predominant mechanism of regulating carotenogenesis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Beyer P: Carotene biosynthesis in daffodile chromoplasts: On the membrane-integral desaturation and cyclization reactions. In: Boyer CD, Shannon JC, Hardison RC (eds) Physiology, Biochemistry and Genetics of Nongreen Plastids. pp. 157–170. American Society of Plant Physiology, Rockville, TN (1989).

    Google Scholar 

  2. Bouvier F, Hugueney P, Dharlingue A, Kuntz M, Camara B: Xanthophyll biosynthesis in chromoplasts: Isolation and molecular cloning of an enzyme catalyzing the conversion of 5,6-epoxycarotenoid into ketocarotenoid. Plant J 6: 45–54 (1994).

    Google Scholar 

  3. Bramley PM: The in vitro biosynthesis of carotenoids. Adv Lipid Res 21: 243–279 (1985).

    Google Scholar 

  4. Bramley PM, Than A, Davies BH: Alternative pathways of carotene cyclization in Phycomyces blakesleeanus. Phytochemistry 16: 235–238 (1977).

    Google Scholar 

  5. Britton G: Biosynthesis of carotenoids. In: Goodwin TW (ed) Plant Pigments, pp. 133–180. Academic Press, New York (1988).

    Google Scholar 

  6. Camara B, Dogbo O: Demonstration and solubilization of lycopene cyclase from Capsicum chromoplast membranes. Plant Physiol 80: 172–184 (1986).

    Google Scholar 

  7. Chamovitz D, Misawa N, Sandmann G, Hirschberg J: Molecular cloning and expression in Escherichia coli of a cyanobacterial gene coding for phytoene synthase, a carotenoid biosynthesis enzyme. FEBS Lett 296: 305–310 (1992).

    Google Scholar 

  8. Cunningham FXJr., Chamovitz D, Misawa N, Gantt E, Hirschberg J: Cloning and functional expression in Escherichia coli of a cyanobacterial gene for lycopene cyclase, the enzyme that catalyzes the biosynthesis of β-carotene. FEBS Lett 328: 130–138 (1993).

    Google Scholar 

  9. Cunningham FXJr., Sun ZR, Chamovitz D, Hirschberg J, Gantt E: Molecular structure and enzymatic function of lycopene cyclase from the cyanobacterium Synechococcus sp strain PCC7942. Plant Cell 6: 1107–1121 (1994).

    Google Scholar 

  10. Davies BH: A novel sequence for phytoene dehydrogenation in Rhodospirillum rubrum. Biochem J 116: 93–99 (1970).

    Google Scholar 

  11. Deruere J, Bouvier, Steppuhn J, Klein A, Camara B, Kuntz M: Structure and expression of two plant genes encoding chromoplast-specific proteins: occurrence of partially spliced transcripts. Biochem Biophys Res Commun 199: 1144–1150 (1994).

    Google Scholar 

  12. Fraser PD, Truesdale MR, Bird CR, Schuch W, Bramley PM: Carotenoid biosynthesis during tomato fruit development. Plant Physiol 105: 405–413 (1994).

    Google Scholar 

  13. Gillaspy G, Ben-David H, Gruissem W: Fruits: a developmental perspective. Plant Cell 5: 1439–1451 (1993).

    Google Scholar 

  14. Guiliano G, Bartley GE, Scolnik P: Regulation of carotenoind biosynthesis during tomato development. Plant Cell 5: 379–387 (1993).

    Google Scholar 

  15. Goodwin TW, Jamikorn M: Biosynthesis of carotenoids in ripening tomatoes. Nature 170: 104–105 (1955).

    Google Scholar 

  16. Grierson D, Schuch W: Control of ripening. Phil Trans R Soc Lond (Biol) 342: 241–250 (1993).

    Google Scholar 

  17. Hundle BS, Obrien DA, Beyer P, Kleinig H, Hearst JE: In vitro expression and activity of lycopene cyclase and β-carotene hydroxylase from Erwinia herbicola. FEBS Lett 315: 329–334 (1993).

    Google Scholar 

  18. Jones BL, Porter JW: Biosynthesis of carotenes in higher plants. CRC Crit Rev Plant Sci 3: 295–324 (1986).

    Google Scholar 

  19. Kuntz M, Römer S, Suire C, Hugueney P, Weil JH, Schantz R, Camara B: Identification of a cDNA for the plastid-located geranylgeranyl pyrophosphate synthase from Capsicum annuum: correlative increase in enzyme activity and transcript level during fruit ripening. Plant J 2: 25–34 (1992).

    Google Scholar 

  20. Kushwaha SC, Subbarayan C, Beeler DA, Porter JW: The conversion of lycopene-15–15′-3H to cyclic carotenes by soluble extracts of higher plant plastids. J Biol Chem 244: 3635–3642 (1969).

    Google Scholar 

  21. Kyte J, Doolittle RF: A simple method for displaying the hydropathic character of a protein. J Mol Biol 157: 105–132 (1982).

    Google Scholar 

  22. Laval-Martin D, Quennement J, Moneger R: Pigment evolution in Lycopersicon esculentum during growth and ripening. Biochemistry 14: 2357–2362 (1975).

    Google Scholar 

  23. Misawa N, Nakagawa M, Kobayashi K, Yamano S, Izawa I, Nakamura K, Harashima K: Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products in Escherichia coli. J Bact 172: 6704–6712 (1990).

    Google Scholar 

  24. Pecker I, Chamovitz D, Linden H, Sandmann G, Hirschberg J: A single polypeptide catalyzing the conversion of phytoene to ζ-carotene is transcriptionally regulated during tomato fruit ripening. Proc Natl Acad Sci USA 89: 4962–4966 (1992).

    Google Scholar 

  25. Ray JA, Moureau P, Bird AS, Grierson D, Maunders M, Truesdale M, Bramley PM, Schuch W: Cloning and characterization of a gene involved in phytoene synthesis from tomato. Plant Mol Biol 19: 401–404 (1992).

    Google Scholar 

  26. Raymundo LC, Griffith AE, Simpson KL: Biosynthesis of carotenoids in tomato fruits. Phytochemistry 9: 1239–1245 (1970).

    Google Scholar 

  27. Raymundo LC, Griffiths AE, Simpson KL. Dimethyl sulfoxide and biosynthesis of carotenoids in detached tomatoes. Phytochemistry 6: 1527–1532 (1967).

    Google Scholar 

  28. Römer S, Hugueney P, Bouvier F, Camara B, Kuntz M: Expression of the genes encoding the early carotenoid biosynthetic enzymes in Capsicum annuum. Biochem Biophys Res Commun 196: 1414–1421 (1993).

    Google Scholar 

  29. Sambrook JF, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory, Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor (1989).

    Google Scholar 

  30. Sandmann G: Phytoene desaturase: genes, enzymes and phylogenetic aspects. J Plant Physiol 143: 444–447 (1994).

    Google Scholar 

  31. Sandmann G, Böger P: Inhibition of carotenoid biosynthesis by herbicides. In: Böger P, Sandmann G (eds) Target Sites of Herbicide Action, pp. 25–44. CRC Press, Boca Raton, FL (1989).

    Google Scholar 

  32. Shaish A, Ben-Amotz A, Avron M: Biosynthesis of β-carotene in Dunaliella. Meth Enzymol 213: 439–444 (1992).

    Google Scholar 

  33. Spurgeon SL, Porter JW: Biosynthesis of Carotenoids. In: Porter JW, Spurgeon SL (eds) Biochemistry of Isoprenoid Compounds, pp. 1–122. Wiley, New York (1980).

    Google Scholar 

  34. Thelander M, Narita JO, Gruissem W: Plastid differentiation and pigment biosynthesis during tomato fruit ripening. Curr Topics Plant Biochem Physiol 5: 128–141 (1986).

    Google Scholar 

  35. To KY, Lai EM, Lee LY, Lin TP, Hung CH, Chen CL, Chang YS, Liu ST: Analysis of the gene cluster encoding carotenoid biosynthesis in Erwinia herbicola EHO13. Microbiology 140: 331–339 (1994).

    Google Scholar 

  36. Tomes ML: Temperature inhibition of carotenoid synthesis in tomato. Bot Gaz 124: 180–185 (1963).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pecker, I., Gabbay, R., Cunningham, F.X. et al. Cloning and characterization of the cDNA for lycopene β-cyclase from tomato reveals decrease in its expression during fruit ripening. Plant Mol Biol 30, 807–819 (1996). https://doi.org/10.1007/BF00019013

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00019013

Key words

Navigation