Research Paper | Published:

Using Antisense RNA to Study Gene Function: Inhibition of Carotenoid Biosynthesis in Transgenic Tomatoes

Bio/Technologyvolume 9pages635639 (1991) | Download Citation



Transgenic tomato plants expressing antisense RNA to a ripening related gene (pTOM5) have yellow ripening fruit and pale colored flowers. The yellow fruit color is correlated with a severe reduction in the level of the pTOMS gene mRNA during fruit ripening. The level of carotenoids in ripening fruit from selected transgenic plants showing yellow fruit was reduced by more than 97 percent. In addition, the carotenoid lycopene, which is primarily responsible for the red color of ripening fruit, was reduced to undetectable levels (<0.1%). These data indicate that the pTOM5 gene is crucial to tomato fruit carotenoid biosynthesis.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1

    van der Krol, A.R., Lenting, P.E., Veenstra, J., van der Meer, I.M., Koes, R.E., Gerats, A.G.M., Mol, J.N.M. and Stuitje, A.R. 1988. An antisense chalcone synthase gene in transgenic plants inhibits flower pigmentation. Nature 333: 866–869.

  2. 2

    Smith, C.J.S., Watson, C.F., Ray, J., Bird, C.R., Morris, P.C., Schuch, W. and Grierson, D. 1988. Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes. Nature 334: 724–726.

  3. 3

    Rodermel, S.R., Abbott, M.S. and Bogorad, L. 1988. Nuclear-organelle interactions: Nuclear antisense gene inhibits ribulose bisphosphate carboxylase enzyme levels in transformed tobacco plants. Cell 55: 673–681.

  4. 4

    Sheehy, R.E., Kramer, M. and Hiatt, W.R. 1988. Reduction of polygalacturonase activity in tomato fruit by antisense RNA. Proc. Natl. Acad. Sci. USA 85: 8805–8809.

  5. 5

    Stockhaus, J., Hofer, M., Renger, G., Westhoff, P., Wydrzynski, T. and Willmitzer, L. 1990. Anti-sense RNA efficiently inhibits formation of the 10 kd polypeptide of photosystem II in transgenic potato plants: analysis of the role of the 10 kd protein. EMBO J. 9: 3013–3021.

  6. 6

    Hamilton, A.J., Lycett, G.W. and Grierson, D. 1990. Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature 346: 284–287.

  7. 7

    Smith, C.J.S., Watson, C.F., Morris, P.C., Bird, C.R., Seymour, G.B., Gray, J.E., Arnold, C., Tucker, G.A., Schuch, W., Harding, S. and Grierson, D. 1990. Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes. Plant Mol. Biol. 14: 369–379.

  8. 8

    Slater, A., Maunders, M.J., Edwards, K., Schuch, W. and Grierson, D. 1985. Isolation and characterisation of cDNA clones for tomato polygalacturonase and other ripening-related proteins. Plant. Mol. Biol. 5: 137–147.

  9. 9

    Mansson, P.-E., Hsu, D. and Stalker, D. 1985. Characterisation of fruit specific cDNAs from tomato. Mol. Gen. Genet. 200: 356–361.

  10. 10

    Lincoln, J.E., Cordes, S., Read, E. and Fischer, R.L. 1987. Regulation of gene expression by ethylene during Lycopersicm esculentum fruit development. Proc. Natl. Acad. Sci. USA 84: 2793–2797.

  11. 11

    Ray, J., Bird, C.R., Maunders, M., Grierson, D. and Schuch, W. 1987. Sequence of pTOM5, a ripening related cDNA from tomato. Nucl. Acids Res. 24: 10587.

  12. 12

    Maunders, M.J., Holdsworth, M.J., Slater, A., Knapp, J., Bird, C.R., Schuch, W. and Grierson, D. 1987. Ethylene stimulates the accumulation of ripening-related mRNAs in tomatoes. Plant, Cell Environ. 10: 177–184.

  13. 13

    Armstrong, G.A., Alberti, M. and Hearst, J.E. 1990. Conserved enzymes mediate the early reactions of carotenoid biosynthesis in nonphotosynthetic and photosynthetic prokaryotes. Proc. Natl. Acad. Sci. USA. 87: 9975–9979.

  14. 14

    Bird, C.R., Smith, C.J.S., Ray, J.A., Moureau, P., Sevan, M.W., Bird, A.S., Hughes, S., Morris, P.C., Grierson, D. and Schuch, W. 1988. The tomato polygalacturonase gene and ripening-specific expression in transgenic plants. Plant Mol. Biol. 11: 651–662.

  15. 15

    Porter, J.W. and Lincoln, R.E. 1950. I. Lycopersicon selections containing a high content of carotenes and colorless polyenes. II. The mechanism of carotenoid biosynthesis. Arch. Biochem. 27: 390–403.

  16. 16

    Knapp, J., Moureau, P., Schuch, W. and Grierson, D. 1989. Organisation and expression of polygalacturonase and other ripening related genes in Ailsa Craig ‘Neverripe’ and ‘Ripening inhibitor’ tomato mutants. Plant Mol. Biol. 12: 105–116.

  17. 17

    Stevens, M.A. and Rick, C.M. 1989. Genetics and breeding, p. 35–109. In: The Tomato Crop. Atherton J. G. and Rudich J. J. (Eds.). Chapman and Hall, New York.

  18. 18

    Armstrong, G.A., Alberti, M., Leach, F. and Hearst, J.E. 1989. Nucleotide sequence, organisation and Nature of the protein products of the carotenoid biosynthesis gene cluster of Rhodobacter capsulatus. Mol. Gen. Genet. 216: 254–268.

  19. 19

    Dogbo, O., Laferriere, A., D'Harlingue, A. and Camara, B. 1988. Carotenoid biosynthesis: Isolation and characterization of a bifunctional enzyme catalyzing the synthesis of phytoene. Proc. Natl. Acad. Sci. USA. 85: 7054–7058.

  20. 20

    Frecknall, E.A. and Pattenden, G. 1984. Carotenoid differences in isogenic lines of tomato fruit colour mutants. Phytochemistry. 23: 1707–1710.

  21. 21

    Darby, L.A., Ritchie, D.B. and Taylor, I.B. 1978. Isogenic lines of the tomato ‘Ailsa Craig’. Ann. Rep. Glasshouse Crops Research Inst. 168–184.

  22. 22

    Kinzer, S.M., Schwager, S.J. and Mutschler, M.A. 1990. Mapping of ripening-related or -specific cDNA clones of tomato. Theor. Appl. Genet. 79: 489–496.

  23. 23

    Bevan, M.W. 1984. Agrobacterium vectors for plant transformation. Nucl. Acids Res. 12: 8711–8721.

  24. 24

    Lichtenstein, C. and Draper, J. 1985. Genetic engineering of plants, p. 67–120. In: DNA Cloning, Vol II (Glover, M. D. (Ed.). IRL Press, Oxford).

  25. 25

    Britton, G. 1985. General carotenoids methods. Methods Enzymol. 111: 113–149.

  26. 26

    Lichtenthaler, H.K. and Wellburn, A.R. 1983. Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem. Soc. Trans. 11: 591–592.

  27. 27

    Davies, B.H. 1976. Carotenoids, p. 38–162. In: Chemistry and Biochemistry of Plant Pigments, Goodwin T. W. (Ed.). Academic Press, London and New York.

Download references

Author information

Author notes

  1. Wolfgang Schuch: Corresponding author.


  1. ICI Seeds, Plant Biotechnology Section, Jealott's Hill Research Station, Bracknell, Berkshire, RG12 6EY, U.K.

    • Colin R. Bird
    • , John A. Ray
    • , Jonathon D. Fletcher
    • , Jeremy M. Boniwell
    • , Alison S. Bird
    • , Chantal Teulieres
    •  & Wolfgang Schuch
  2. Department of Biochemistry, Royal Holloway and Bedford New College, University of London, Egham, Surrey, TW20 0EX, U.K.

    • Ian Blain
    •  & Peter M. Bramley


  1. Search for Colin R. Bird in:

  2. Search for John A. Ray in:

  3. Search for Jonathon D. Fletcher in:

  4. Search for Jeremy M. Boniwell in:

  5. Search for Alison S. Bird in:

  6. Search for Chantal Teulieres in:

  7. Search for Ian Blain in:

  8. Search for Peter M. Bramley in:

  9. Search for Wolfgang Schuch in:

About this article

Publication history



Issue Date


Further reading