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Isolation of a pea (Pisum sativum) seed lipoxygenase promoter by inverse polymerase chain reaction and characterization of its expression in transgenic tobacco

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Abstract

Part of the 5′-flanking sequence of a pea (Pisum sativum) lipoxygenase (LOX) gene was cloned, after amplification from genomic DNA by inverse polymerase chain reaction. Translational and transcriptional fusions of 818 bp of the 5′-flanking region and its deletion derivatives (−513 and −356) were made to a β-glucuronidase (GUS)-coding sequence and introduced into tobacco. Analysis of T1 transformants showed that the 818 bp 5′-flanking sequence drove GUS expression in seeds that was temporally regulated in a fashion similar to the accumulation of LOX mRNA in developing pea seeds. Contrary to expectations, however, expression of the 818 bp promoter-GUS fusion was not seed-specific; GUS activity was highest in leaves and also present in stems and, to a lesser extent, roots. Deletion analysis identified the region between −818 and −513 as essential for high-level, temporally regulated expression in seeds and also indicated that the sequence between −513 and −356 plays a negative role in leaf/stem, but not seed, expression. Comparison of translational and transcriptional fusions indicated that the LOX initiation codon was used more efficiently than the GUS initiation codon by the tobacco leaf translational apparatus.

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References

  1. Bäumlein H, Nagy I, Villarroel R, Inze D, Wobus U: Cis-analysis of a seed protein gene promoter: the conservative RY repeat CATGCATG within the legumin box is essential for tissue-specific expression of a legumin gene. Plant J 2: 233–239 (1992).

    Google Scholar 

  2. Bell E, Mullett JE: Lipoxygenase gene expression is modulated in plants by water deficit, wounding and methyl jasmonate. Mol Gen Genet 230: 456–462 (1991).

    Google Scholar 

  3. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).

    Google Scholar 

  4. Bustos MM, Guiltinan MJ, Jordano J, Begum F, Kalkan A, Hall TC: Regulation of β-glucuronidase expression in transgenic tobacco plants by an AT-rich, cis-acting sequence found upstream of a French bean β-phaseolin gene. Plant Cell 1: 839–853 (1989).

    Google Scholar 

  5. Casey R, Domoney C, Nielsen NC: Isolation of a cDNA clone for pea (Pisum sativum) seed lipoxygenase. Biochem J 232: 79–85 (1985).

    Google Scholar 

  6. Chamberland S, Daigle N, Bernier F: The legumin boxes and the 3′ part of a soybean β-conglycinin promoter are involved in seed gene expression in transgenic tobacco plants. Plant Mol Biol 19: 937–949 (1992).

    Google Scholar 

  7. Creelman RA, Bell E, Mullett JE: Involvement of a lipoxygenase-like enzyme in abscisic acid biosynthesis. Plant Physiol 99: 1258–1260 (1992).

    Google Scholar 

  8. Croft KPC, Juttner F, Slusarenko AJ: Volatile products of the lipoxygenase pathway evolved from Phaseolus vulgaris (L.) leaves inoculated with Pseudomonas syringae pv. phaseolicola. Plant Physiol 101: 13–24 (1993).

    Google Scholar 

  9. Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: version II. Plant Mol Biol Rep 1: 19–21 (1983).

    Google Scholar 

  10. Dickinson CD, Evan RP, Nielsen NC: RY repeats are conserved in the 5′ flanking region of legume seed protein genes. Nucl Acids Res 16: 371 (1988).

    Google Scholar 

  11. Domoney C, Casey R, Turner L, Ellis N: Pisum lipoxygenase genes. Theor Appl Genet 81: 800–805 (1991).

    Google Scholar 

  12. Domoney C, Firmin JL, Sidebottom C, Ealing PM, Slabas A, Casey R: Lipoxygenase heterogeneity in Pisum sativum. Planta 181: 35–43 (1990).

    Google Scholar 

  13. Ealing PM, Casey R: The complete amino acid sequence of a pea (Pisum sativum) seed lipoxygenase predicted from a near full-length cDNA. Biochem J 253: 915–918 (1988).

    Google Scholar 

  14. Ealing PM, Casey R: The cDNA cloning of a pea (Pisum sativum) seed lipoxygenase. Sequence comparisons of the two major seed lipoxygenase isoforms. Biochem J 264: 929–932 (1989).

    Google Scholar 

  15. Farmer EE, Ryan CA: Octadecanoid precursors of jasmonic acid activate the synthesis of wound-inducible proteinase inhibitors. Plant Cell 4: 129–134 (1992).

    Google Scholar 

  16. Fiedler U, Filistein R, Wobus U, Bäumlein H: A complex ensemble of cis-regulatory elements controls the expression of a Vicia faba non-storage seed protein gene. Plant Mol Biol 22: 669–679 (1993).

    Google Scholar 

  17. Frazier PJ: Lipoxygenase action and lipid binding in breadmaking. Bakers' Digest 53: 8–29 (1979).

    Google Scholar 

  18. Futers TS, Onde S, Turet M, Cuming AC: Sequence analysis of the tandemly linked Em genes from wheat. Plant Mol Biol 23: 1067–1072 (1993).

    Google Scholar 

  19. Ghosh D: A relational database of transcription factors. Nucl Acids Res 18: 1749–1756 (1990).

    Google Scholar 

  20. Goldberg RB: Regulation of plant gene expression. Phil Trans Roy Soc B 314: 343–353 (1986).

    Google Scholar 

  21. Green PJ, Kay SA, Chua N-M: Sequence specific interactions of a pea nuclear factor with light-responsive elements upstream of the rbcs-3A gene. EMBO J 6: 2543–2549 (1987).

    Google Scholar 

  22. Green PJ, Yong M-H, Cuozzo M, Kano-Murakami Y, Silverstein P, Chua N-H: Binding site requirements for pea nuclear protein factor GT-1 correlate with sequences required for light dependent transcriptional activation of the rbcs-3A gene. EMBO J 7: 4035–4044 (1988).

    Google Scholar 

  23. Hatanaka A: The biogenesis of green odour by green leaves. Phytochemistry 34: 1201–1218 (1993).

    Google Scholar 

  24. Henskens LAM, Fornerod MWJ, Rueb S, Winkler AA, van der Veen S, Schilperoort RA: Translation controls the expression level of a chimaeric reporter gene. Plant Mol Biol 20: 921–938 (1992).

    Google Scholar 

  25. Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).

    Google Scholar 

  26. Jordano J, Almoguera C, Thomas TL: A sunflower helianthinin gene upstream ensemble contains an enhancer and sites of nuclear protein interaction. Plant Cell 1: 855–866 (1989).

    Google Scholar 

  27. Kato T, Shirano Y, Kawazu T, Tada Y, Itoh E, Shibata D: A modified β-glucuronidase gene: sensitive detection of plant promoter activities in suspension cultured cells of tobacco and rice. Plant Mol Biol Rep 9: 333–339 (1991).

    Google Scholar 

  28. Keovahong P, Thilly W: Fidelity of DNA polymerases in DNA amplification. Proc Natl Acad Sci USA 86: 9253–9257 (1989).

    Google Scholar 

  29. Kreis M, Shewry PR, Forde BG, Forde J, Miflin BJ: Structure and evolution of seed storage proteins and their genes with particular reference to those of wheat, barley and rye. Oxford Surv Plant Mol Cell Biol 2: 253–317 (1985).

    Google Scholar 

  30. Lee D, Turner L, Davies DR, Ellis THN: An RFLP marker for rb in pea. Theor Appl Genet 75: 362–365 (1988).

    Google Scholar 

  31. Lelievre JM, Oliveira LO, Nielsen NC: 5′ CATG-CATG 3′ elements modulate the expression of glycinin genes. Plant Physiol 98: 387–391 (1992).

    Google Scholar 

  32. Lessard PA, Allen RD, Fujiwara T, Beachy RN: Upstream regulatory sequences from two β-conglycinin genes. Plant Mol Biol 22: 873–885 (1993).

    Google Scholar 

  33. Melan MA, Dong X, Endara ME, Davis KR, Ausubel FM, Peterman TK: An Arabidopsis thaliana lipoxygenase gene can be induced by pathogens, abscisic acid, and methyl jasmonate. Plant Physiol 101: 441–450 (1993).

    Google Scholar 

  34. Mullineaux PM, Guerineau F, Accotto G-P: Processing of complementary sense RNAs of Digitaria streak virus in its host and in transgenic tobacco. Nucl Acids Res 18: 7259–7265 (1990).

    Google Scholar 

  35. Nap J-P, Keizer P, Jansen R: First-generation transgenic plants and statistics. Plant Mol Biol Rep 11: 156–164 (1993).

    Google Scholar 

  36. Ochman H, Gerber AS, Hartl DL: Genetic applications of an inverse polymerase chain reaction. Genetics 120: 621–623 (1988).

    Google Scholar 

  37. Parry AD, Horgan R: Carotenoid metabolism and the biosynthesis of abscisic acid. Phytochemistry 30: 815–821 (1991).

    Google Scholar 

  38. Rao AG, Flynn P: Microtiter plate-based assay for β-D-glucuronidase: a quantitative approach. In: Gallagher SR (ed) GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression, pp. 89–99. Academic Press, New York (1992).

    Google Scholar 

  39. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  40. Schindler U, Cashmore AR: Photoregulated gene expression may involve ubiquitous DNA binding proteins. EMBO J 9: 3415–3427 (1990).

    Google Scholar 

  41. Steel RGD, Torrie JH: Principles and Procedures of Statistics, pp. 107–109. McGraw-Hill, New York (1960).

    Google Scholar 

  42. Thomas MS, Flavell RB: Identification of an enhancer element for the endosperm- specific expression of high molecular weight glutenin. Plant Cell 2: 1171–1180 (1990).

    Google Scholar 

  43. Warner SAJ, Scott R, Draper J: Isolation of an asparagus intracellular PR gene (AoPR1) wound-responsive promoter by the inverse polymerase chain reaction and its characterization in transgenic tobacco. Plant J 3: 191–201 (1993).

    Google Scholar 

  44. Yoon S, Klein BP: Some properties of pea lipoxygenase isoenzymes. J Agric Food Chem 27: 955–962 (1979).

    Google Scholar 

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Author notes

  1. Shaun L. A. Hobbs

    Present address: CAB International, OX10 8DE, Wallingford, UK

Authors and Affiliations

  1. John Innes Centre, Colney Lane, NR4 7UH, Norwich, UK

    Colette Forster, Eddie Arthur, Phil Mullineaux & Rod Casey

  2. Istituto di Fitovirologia Applicata, Strada delle Cacce, 10135, Torino, Italy

    Stefania Crespi

  3. Plant Biotechnology Institute, National Research Council of Canada, S79 OW9, Saskatoon, Saskatchewan, Canada

    Shaun L. A. Hobbs

Authors
  1. Colette Forster
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  2. Eddie Arthur
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  3. Stefania Crespi
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  4. Shaun L. A. Hobbs
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  5. Phil Mullineaux
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  6. Rod Casey
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Forster, C., Arthur, E., Crespi, S. et al. Isolation of a pea (Pisum sativum) seed lipoxygenase promoter by inverse polymerase chain reaction and characterization of its expression in transgenic tobacco. Plant Mol Biol 26, 235–248 (1994). https://doi.org/10.1007/BF00039535

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  • DOI: https://doi.org/10.1007/BF00039535

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