Plant Molecular Biology

, Volume 26, Issue 4, pp 1073–1084 | Cite as

Structure of the tomato Adh2 gene and Adh2 pseudogenes, and a study of Adh2 gene expression in fruit

  • Terry Longhurst
  • Elizabeth Lee
  • Rick Hinde
  • Colin Brady
  • Jim Speirs
Research Article

Abstract

A cDNA library was constructed from RNA from the pericarp of ripe tomato fruit and four cDNAs encoding ADH2 were isolated and characterized. The cDNAs encode a peptide 379 amino acids in length. They hybridized strongly with a 1.8 kb RNA species well represented in RNA from ripe, but not from mature, unripe fruit, and strongly to a similar RNA species present in hypoxic, but not in aerobic roots. Northern analysis showed that the mRNA for ADH2 in fruit increased in abundance through ripening, particularly during late ripening. In pericarp tissue of fruit, the Adh2 mRNA level increased to a maximum within 8–16 h of exposure to atmospheres with 3% (v/v) oxygen, and returned to the basal level within 16 h of a return to air. The mRNA level was sensitive to the oxygen level in the atmosphere, increasing 20-fold in 12% (v/v) oxygen and 100-fold in 3% oxygen.

The homologous tomato Adh2 gene was isolated from a genomic library. The gene has an overall length of 2334 bp from transcription start site to poly(A) addition site and includes eight introns.

Southern blot analysis of tomato genomic DNA identified multiple Adh2-related sequences. Two of these, PSA1 and PSA2, were cloned and found to have 94% similarity with each other and 77% similarity with the tomato Adh2 gene over a 1000 bp region. The homologous regions include introns and exons but the equivalent exons contain frame shifts, deletions and stop codons. The two regions are therefore presumptive pseudogenes.

Key words

ADH2 cDNA fruit gene hypoxia ripening 

Abbreviations

ADH

alcohol dehydrogenase

Adh

pertaining to an ADH gene

ARE

anaerobic responsive element

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Bicsak TA, Kann LR, Reiter A, Chase T: Tomato alcohol dehydrogenase: purification and substrate specificity. Arch Biochem Biophys 216: 605–615 (1982).PubMedGoogle Scholar
  2. 2.
    Brady CJ: Fruit ripening. Annu Rev Plant Physiol 38: 155–178 (1987).Google Scholar
  3. 3.
    Breathnach R, Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem 50: 349–383 (1981).CrossRefPubMedGoogle Scholar
  4. 4.
    Chang C, Meyerowitz EM: Molecular cloning and DNA sequence of the Arabidopsis thaliana alcohol dehydrogenase gene. Proc Natl Acad Sci USA 83: 1408–1412 (1986).PubMedGoogle Scholar
  5. 5.
    Cushman JC, Bohnert HJ: Salt stress alters A/T-rich DNA-binding factor interactions within the phospho-enolpyruvate carboxylase promoter from Mesembryanthemum crystallinum. Plant Mol Biol 20: 411–424 (1992).PubMedGoogle Scholar
  6. 6.
    Dean C, Tamaki S, Dunsmuir P, Favreau M, Katayama C, Dooner H, Bedbrook J: mRNA transcripts of several plant genes are polyadentylated at multiple sites in vivo. Nucl Acids Res 14: 2229–2240 (1986).PubMedGoogle Scholar
  7. 7.
    Dennis ES, Sachs MM, Gerlach WL, Finnegan EJ, Peacock JW: Molecular analysis of the alcohol dehydrogenase 2 (ADH2) gene of maize. Nucl Acids Res 13: 727–743 (1985).PubMedGoogle Scholar
  8. 8.
    Dennis ES, Gerlach WL, Walker JC, Lavin M, Peacock WJ: Anaerobically regulated aldolase gene of maize: a chimaeric origin? J Mol Biol 202: 759–767 (1988).PubMedGoogle Scholar
  9. 9.
    Ferl RJ, Laughner BH: In vivo detection of regulatory factor binding sites of Arabidopsis thaliana Adh. Plant Mol Biol 12: 357–366 (1989).CrossRefGoogle Scholar
  10. 10.
    Fourney RM, Miyakoshi J, Day RS, Paterson MC: Northern blotting: efficient RNA staining and transfer. Focus 10 (1): 5–9 (1988).Google Scholar
  11. 11.
    Genez AL, Staraci LC, Alexander DC, Rejda JM, Williamson VM, Chase TJnr, Williams BG: Isolation of a tomato alcohol dehydrogenase 2-encoding cDNA using phage-promoted antibody screening of a plasmid cDNA library. GenBank database, accession number M86724 (1992).Google Scholar
  12. 12.
    Genez AL, Staraci LC, Alexander DC, Rejda JM, Williamson VM, Chase TJnr, Williams BG: Isolation of a tomato alcohol dehydrogenase 2-encoding cDNA using phage-promoted antibody screening of a plasmid cDNA library. Gene 123: 157–164 (1993).CrossRefPubMedGoogle Scholar
  13. 13.
    Gerlach WL, Pryor AJ, Dennis ES, Ferl AJ, Sacha MM, Peacock WJ: cDNA cloning and induction of the alcohol dehydrogenase gene (ADH 1) of maize. Proc Natl Acad Sci USA 79: 2981–2985 (1982).Google Scholar
  14. 14.
    Joshi CP: An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucl Acids Res 15: 6643–6653 (1987).PubMedGoogle Scholar
  15. 15.
    Kloesgen RB, Gierl A, Schwarz-Sommer Z, Saedler H: Molecular analysis of the waxy-locus of Zea mays. Mol Gen Genet 203: 237–244 (1986).CrossRefGoogle Scholar
  16. 16.
    Lee E, Speirs J, McGlasson WB, Brady CJ: Messenger RNA changes in tomato fruit pericarp in response to propylene, wounding or ripening. J Plant Physiol 129: 287–299 (1987).Google Scholar
  17. 17.
    Lincoln JE, Fischer RL: Diverse mechanisms for the regulation of ethylene-inducible gene expression. Mol Gen Genet 212: 71–75 (1988).CrossRefPubMedGoogle Scholar
  18. 18.
    Llewellyn DJ, Finnegan EJ, Ellis JG, Dennis ES, Peacock WJ. Structure and expression of an alcohol dehydrogenase 1 gene from Pisum sativum (cv. ‘Greenfeast’). J Mol Biol 195: 115–123 (1987).PubMedGoogle Scholar
  19. 19.
    Longhurst TJ, Tung HF, Brady CJ: Developmental regulation of the expression of alcohol dehydrogenase in ripening tomato fruit. J Food Biochem 14: 421–433 (1990).Google Scholar
  20. 20.
    Lyons JM, Pratt HK: Effect of stage of maturity and ethylene treatment on respiration and ripening of tomato fruits. Proc Am Soc Hort Sci 84: 491–500 (1964).Google Scholar
  21. 21.
    Matton DP, Brisson N: Nucleotide sequence of two potato alcohol dehydrogenase cDNAs. Nucl Acids Res 18: 3070 (1990).PubMedGoogle Scholar
  22. 22.
    Matton DP, Constabel P, Brisson N: Alcohol dehydrogenase gene expression in potato following elicitor and stress treatment. Plant Mol Biol 14: 775–783 (1990).CrossRefPubMedGoogle Scholar
  23. 23.
    Messing J, Geraghty D, Heidecker G, Hnu N-T, Kridl J, Rubenstein I: Plant gene structure. In: Kosuge T, Meredith CP, Hollaender A (eds) Genetic Engineering of Plants: An Agricultural Perspective, pp. 211–227. Plenum Press, New York (1983).Google Scholar
  24. 24.
    Mozer TJ: Partial purification and characterisation of the mRNA for α-amylase from barley aleurone layers. Plant Physiol 65: 834–837 (1980).Google Scholar
  25. 25.
    Palmiter RD: Ovalbumin messenger ribonucleic acid translation. Comparable rates of polypeptide initiation and elongation on ovalbumin and globin messenger ribonucleic acid in rabbit reticulocyte lysate. Biol Chem 248: 2095–2106 (1973).Google Scholar
  26. 26.
    Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).Google Scholar
  27. 27.
    Sanger F, Niklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).PubMedGoogle Scholar
  28. 28.
    Speirs J, Brady CJ: A coordinated decline in the synthesis of subunits of ribulosebisphosphate carboxylase in aging wheat leaves. II. Abundance of messenger RNA. Aust J Plant Physiol 8: 603–618 (1981).Google Scholar
  29. 29.
    Tanksley SD: Linkage, chromosomal association and expression of Adh-1 and Pgm-2 in tomato. Biochem Genet 17: 1159–1167 (1979).PubMedGoogle Scholar
  30. 30.
    Tanksley SD, Jones RA: Effects of O2 stress on tomato alcohol dehydrogenase activity: description of a second ADH coding gene. Biochem Genet 19: 397–409 (1981).PubMedGoogle Scholar
  31. 31.
    Thomas MR, Matsumoto S, Cain P, Scott NS: Repetitive DNA of grapevine: classes present and sequences suitable for cultivar identification. Theor Appl Genet 86: 173–180 (1993).Google Scholar
  32. 32.
    Tihanyi K, Fontanell A, Thirion J: Gene regulation during anaerobiosis in soya roots. Biochem Genet 27: 719–730 (1989).PubMedGoogle Scholar
  33. 33.
    Van der Straeten D, Pousada RA, Gielen J, Van Montagu M: Tomato alcohol dehydrogenase. Expression during fruit ripening and under hypoxic conditions. FEBS Lett 295: 39–42 (1991).PubMedGoogle Scholar
  34. 34.
    Walker JC, Howard EA, Dennis ES, Peacock WJ: DNA sequences required for anaerobic expression of the maize alcohol dehydrogenase 1 gene. Proc Natl Acad Sci USA 84: 6624–6628 (1987).Google Scholar
  35. 35.
    Werr W, Frommer WB, Maas C, Starlinger P: Structure of the sucrose synthase gene on chromosome 9 of Zea mays L. EMBO J 4: 1373–1380 (1985).Google Scholar
  36. 36.
    Wisman E, Koornneef M, Chase T, Lifshytz E, Ramanna MS, Zabel P. Genetic and molecular characterization of an Adh-1 null mutant in tomato. Mol Gen Genet 226: 120–128 (1991).CrossRefPubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Terry Longhurst
    • 1
  • Elizabeth Lee
    • 1
  • Rick Hinde
    • 2
  • Colin Brady
    • 1
  • Jim Speirs
    • 1
  1. 1.CSIRO Division of Horticulture (Sydney)North RydeAustralia
  2. 2.School of Biological SciencesMacquarie UniversityAustralia

Personalised recommendations