Plant Molecular Biology

, Volume 37, Issue 6, pp 1001–1011 | Cite as

Separation of cis elements responsive to ethylene, fruit development, and ripening in the 5′-flanking region of the ripening-related E8 gene

  • Jill Deikman
  • Ruiling Xu
  • Michelle L. Kneissl
  • Joseph A. Ciardi
  • Kyung-Nam Kim
  • Dan Pelah
Article

Abstract

The E8 gene is expressed at a high level during fruit ripening, and is transcriptionally activated by ethylene. We have identified a 428 bp fragment of the E8 5'-flanking region, from -1528 to -1100, that makes a minimal 35S promoter responsive to ethylene. This fragment confers ethylene-responsiveness only in the 5';-to-3'; orientation; in the reverse orientation it results in increased expression in unripe fruit. Interestingly, this ethylene-responsive construct does not have high levels of expression during fruit ripening, indicating that sequences required for high level expression during fruit ripening are separate from sequences required for ethylene response. The ethylene-responsive sequences of the E8 5';-flanking region interact with the same DNA-binding protein that interacts with sequences required for ethylene responsiveness of the coordinately regulated E4 gene. We also conducted experiments to test the function of a second DNA-binding protein that interacts with both E4 and E8 5';-flanking sequences, the E4/E8-binding protein (E4/E8BP). We examined the effect of an internal deletion from -1088 to -863, which includes the binding site for E4/E8BP, on gene expression. This deletion did not affect expression in ripening fruit, and did not impair ethylene responsiveness. The deletion had a negative effect on expression in unripe fruit, but resulted in increased expression in leaves. These results suggest that the E4/E8BP is not critical for high levels of expression during fruit ripening or for ethylene response, but may play a role in organ-specific gene transcription.

ethylene fruit ripening Lycopersicon esculentum promoter analysis 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Brady CJ, Speirs J: Ethylene in fruit ontogeny and abscission. In Mattoo AK, Suttle JC (eds) The Plant Hormone Ethylene, pp. 235–258. CRC Press, Boca Raton, FL (1991).Google Scholar
  2. 2.
    Cordes S, Deikman J, Margossian LJ, Fischer RL: Interaction of a developmentally regulated DNA-binding factor with sites flanking two different fruit-ripening genes from tomato. Plant Cell 1: 1025–1034 (1989).CrossRefPubMedGoogle Scholar
  3. 3.
    Coupe SA, Deikman J: Characterization of a DNA-binding protein that interacts with 50-flanking regions of two fruitripening genes. Plant J 11: 1207–1218 (1997).PubMedGoogle Scholar
  4. 4.
    Deikman J, Fischer RL: Interaction of a DNA binding factor with the 50-flanking region of an ethylene-responsive fruit ripening gene from tomato. EMBO J 7: 3315–3320 (1988).PubMedGoogle Scholar
  5. 5.
    Deikman J, Kline R, Fischer RL: Organization of ripening and ethylene regulatory regions in a fruit-specific promoter from tomato (Lycopersicon esculentum). Plant Physiol 100: 2013–2017 (1992).Google Scholar
  6. 6.
    DellaPenna D, Lincoln JE, Fischer RL, Bennett AB: Transcriptional analysis of polygalacturonase and other ripening associated genes in Rugters, rin, nor, and Nr tomato fruit. Plant Physiol 90: 1372–1377 (1989).Google Scholar
  7. 7.
    Fang R-X, Nagy F, Sivasubramaniam S, Chua N-H: Multiple cis regulatory elements for maximal expression of the cauliflower mosaic virus 35S promoter in transgenic plants. Plant Cell 1: 141–150 (1989).CrossRefPubMedGoogle Scholar
  8. 8.
    Fray RG, Grierson D: Molecular genetics of tomato fruit ripening. Trends Genet 9: 438–443 (1993).PubMedGoogle Scholar
  9. 9.
    Giovannoni JJ, DellaPenna D, Bennett AB, Fischer RL: Expression of a chimeric polygalacturonase gene in transgenic rin (ripening inhibitor) tomato fruit results in polyuronide degradation but not fruit softening. Plant Cell 1: 53–63 (1989).PubMedGoogle Scholar
  10. 10.
    Good X, Kellogg JA, Wagoner W, Langhoff D, Matsumura W, Bestwick RK: Reduced ethylene synthesis by transgenic tomatoes expressing S-adenosylmethionine hydrolase. Plant Mol Biol 26: 781–790 (1994).PubMedGoogle Scholar
  11. 11.
    Gray J, Picton S, Shabbeer J, Schuch W, Grierson D: Molecular biology of fruit ripening and its manipulation with antisense genes. Plant Mol Biol 19: 69–87 (1992).PubMedGoogle Scholar
  12. 12.
    Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).Google Scholar
  13. 13.
    Klee HJ: Ripening physiology of fruit from transgenic tomato (Lycopersicon esculentum) plants with reduced ethylene synthesis. Plant Physiol 102: 911–916 (1993).PubMedGoogle Scholar
  14. 14.
    Kneissl ML, Deikman J: The tomato E8 gene influences ethylene biosynthesis in fruit but not in flowers. Plant Physiol 112: 537–547 (1996).PubMedGoogle Scholar
  15. 15.
    Kosugi S, Ohashi Y, Nakajima K, Arai Y: An improved assay for β-glucuronidase in transformed cells: methanol almost completely represses a putative endogenous b-glucuronidase activity. Plant Sci 70: 133–140 (1990).CrossRefGoogle Scholar
  16. 16.
    Lam E, Benfey PN, Gilmartin PM, Fang R-X, Chua N-H: Site-specific mutations alter in vitro factor binding and change promoter expression pattern in transgenic plants. Proc Natl Acad Sci USA 86: 7890–7894 (1989).PubMedGoogle Scholar
  17. 17.
    Lincoln JE, Cordes S, Read E, Fischer RL: Regulation of gene expression by ethylene during Lycopersicon esculentum (tomato) fruit ripening. Proc Natl Acad Sci USA 84: 2793–2797 (1987).PubMedGoogle Scholar
  18. 18.
    Lincoln JE, Fischer RL: Diverse mechanisms for the regulation of ethylene-inducible gene expression. Mol Gen Genet 212: 71–75 (1988).CrossRefPubMedGoogle Scholar
  19. 19.
    Montgomery J, Goldman S, Deikman J, Margossian L, Fischer RL: Identification of an ethylene-responsive region in the promoter of a fruit ripening gene. Proc Natl Acad Sci USA 90: 5939–5943 (1993).PubMedGoogle Scholar
  20. 20.
    Oeller PW, Min-Wong L, Taylor L, Pike DA, Theologis A: Reversible inhibition of tomato fruit senescence by antisense RNA. Science 254: 437–439 (1991).PubMedGoogle Scholar
  21. 21.
    Ow DW, Wood KV, DeLuca M, deWet JR, Helinski DR, Howell SH: Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science 234: 856–859 (1986).Google Scholar
  22. 22.
    Penarrubia L, Aguilar M, Margossian L, Fischer RL: An antisense gene stimulates ethylene hormone production during tomato fruit ripening. Plant Cell 4: 681–687 (1992).CrossRefPubMedGoogle Scholar
  23. 23.
    Picton S, Barton SL, Bouzayen M, Hamilton AJ, Grierson D: Altered fruit ripening and leaf senescence in tomatoes expressing an antisense ethylene-forming enzyme transgene. Plant J 3: 469–481 (1993).Google Scholar
  24. 24.
    Qin X-F, Holuigue L, Horvath DM, Chua N-H: Immediate early transcription activation by salicyclic acid via the cauliflower mosaic virus as-1 element. Plant Cell 6: 863–874 (1994).CrossRefPubMedGoogle Scholar
  25. 25.
    Rhodes MJC: The maturation and ripening of fruits. In: Thimann KV (ed) Senescence in Plants, pp. 157–205. CRC Press, Boca Raton, FL (1980).Google Scholar
  26. 26.
    Rottmann WH, Peter GF, Oeller PW, Keller JA, Shen NF, Nagy BP, Taylor LP, Campbell AD, Theologis A: 1-Aminocyclopropane-1-carboxylate synthase in tomato is encoded by a multigene family whose transcription is induced during fruit and flower senescence. J Mol Biol 222: 937–961 (1991).CrossRefPubMedGoogle Scholar
  27. 27.
    Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Plainview, NY (1989).Google Scholar
  28. 28.
    Theologis A, Oeller PW, Wong L-M, Rottmann WH, Gantz DM: Use of a tomato mutant constructed with reverse genetics to study fruit ripening, a complex developmental process. Devel Genet 14: 282–295 (1993).Google Scholar
  29. 29.
    Van Haaren MJJ, Houck CM: Strong negative and positive regulatory elements contribute to the high-level fruit-specific expression of the tomato 2A11 gene. Plant Mol Biol 17: 615–630 (1991).PubMedGoogle Scholar
  30. 30.
    Wilkinson JQ, Lanahan MB, Yen H-C, Giovannoni JJ, Klee HJ: An ethylene-inducible component of signal transduction encoded by Never-ripe. Science 270: 1807–1809 (1995).PubMedGoogle Scholar
  31. 31.
    Xu R, Goldman S, Coupe S, Deikman J: Ethylene control of E4 transcription during tomato fruit ripening involves two cooperative cis-elements. Plant Mol Biol 31: 1117–1127 (1996).PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Jill Deikman
    • 1
    • 2
    • 4
  • Ruiling Xu
    • 1
    • 3
  • Michelle L. Kneissl
    • 2
  • Joseph A. Ciardi
    • 5
  • Kyung-Nam Kim
    • 2
  • Dan Pelah
    • 1
    • 6
  1. 1.Department of Biology208 Mueller LaboratoryUSA
  2. 2.Intercollege Graduate Program in Plant PhysiologyUSA
  3. 3.Blood Systems FoundationTempeUSA
  4. 4.Monsanto Company AA2GSt. LouisUSA
  5. 5.Department of HorticulturePennsylvania State UniversityTempeUSA
  6. 6.Ben-Gurion University of the NegevSede Boqer CampusIsrael

Personalised recommendations