Skip to main content
SpringerLink
Log in

The expression of an abundant transmitting tract-specific endoglucanase (Sp41) is promoter-dependent and not essential for the reproductive physiology of tobacco

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

In angiosperms the interactions between the secretory matrix of the stylar transmitting tract and the growing pollen tubes have central roles in determining a successful fertilization. Sp41 is a major glycosylated component of the soluble proteins of the transmitting tract matrix and exhibits (1–3)-β-glucanase activity. It is a member of the pathogenesis-related protein superfamily, but shows developmental regulation as opposed to pathogen induction. In order to investigate the mechanisms regulating Sp41 expression, we isolated and characterized genomic clones corresponding to the sp41 a gene. Sp41 a contains an intervening sequence localized between the sequences encoding for a putative signal peptide and the mature protein. A fragment of 2.5 kb that lies 5′ to the coding region of the gene was sufficient to confer transmitting tract specific expression to a β-glucuronidase reporter gene in transgenic tobacco plants. The sp41 transcripts have unusually long 5′-untranslated sequences. The leader sequences contain small open reading frames, include secondary structures, and may be involved in post-transcriptional regulation. A possible function for Sp41 in reproductive physiology was tested by monitoring tobacco plants transformed with antisense stylar sp41a RNA. Transgenic antisense plants with immunologically and enzymatically undetectable levels of (1–3)-β-glucanase were obtained and their offspring analyzed. The progeny plants did not show any detectable phenotypic modifications as they had a normal flower morphology and were fully fertile.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. An G: Binary Ti vectors for plant transformation and promoter analysis. Meth Enzymol 153: 292–305 (1987).

    Google Scholar 

  2. Ausubel FM, Brent R, Kingston RE, Moore DD, Smith JA, Seidman JG, Struhl K: Current Protocols in Molecular Biology. John Wiley, New York (1989).

    Google Scholar 

  3. Benfey PN, Chua NH: The Cauliflower Mosaic Virus 35S promoter: combinatorial regulation of transcription in plants. Science 250: 959–966 (1990).

    Google Scholar 

  4. Bucciaglia PA, Smith AG: Cloning and characterization of Tag 1, a tobacco anther β-1,3-glucanase expressed during tetrad dissolution. Plant Mol Biol 24: 903–914 (1994).

    Google Scholar 

  5. Budelier KA, Smith AG, Gasser CS: Regulation of a stylar transmitting tissue-specific gene in wild-type and transgenic tomato and tobacco. Mol Gen Genet 224: 183–192 (1990).

    Google Scholar 

  6. Castresana C, deCarvalho F, Gheysen G, Habets M, Inzé D, VanMontagu M: Tissue-specific and pathogen-induced regulation of a Nicotiana plumbaginifolia β-1,3-glucanase gene. Plant Cell 2: 1131–1143 (1990).

    Google Scholar 

  7. Chen C-G, Cornish EC, Clarke AE: Specific expression of an extensin-like gene in the style of Nicotiana alata. Plant Cell 4: 1053–1062 (1992).

    Google Scholar 

  8. Cheung AY, May B, Kawata EE, Gu Q, Wu H-M: Characterization of cDNAs for stylar transmitting tissue-specific proline-rich proteins in tobacco. Plant J 3: 151–160 (1993).

    Google Scholar 

  9. Cohen Y, Niderman T, Mösinger E, fluhr R: β-aminobutyric acid induces the accumulation of pathogenesis-related proteins in tomato (Lycopersicum esculentum L.) plants and resisntance to late blight infection caused by Phytophthora infestans. Plant Physiol 104: 59–66 (1994).

    Google Scholar 

  10. Coleman CE, Kao T-h: The flanking regions of two Petunia inflata S alleles are heterogeneous and contain repetitive sequences. Plant Mol Biol 18: 725–737 (1992).

    Google Scholar 

  11. DamianiJr RD, Wessler SR: An upstream open reading frame represses expression of Lc, a member of the R/B family of maize transcription activators. Proc Natl Acad Sci USA 90: 8244–8248 (1993).

    Google Scholar 

  12. Davies B, Schwarz-Sommer Z: Control of floral organ identity by homeotic MADS-box transcription factors. In: Nover L (ed) Plant Promoters and Transcription Factors, pp. 235–258. Springer-Verlag, Berlin/Heidelberg/New York (1994).

    Google Scholar 

  13. Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).

    Google Scholar 

  14. Donald RGK, Cashmore AR: Mutation of either G box or I box sequences profoundly affects expression from the Arabidopsis rbcS-1A promoter. EMBO J 9: 1717–1726 (1990).

    Google Scholar 

  15. Dzelzkalns VA, Nasrallah JB, Nasrallah ME: Cell-cell communication in plants: self-incompatibility in flower development. Devel Biol 153: 70–82 (1992).

    Google Scholar 

  16. Dzelzkalns VA, Thorsness MK, dwyer KG, Baxter JS, Balent MA, Nasrallah ME, Nasrallah JB: Distinct cisacting elements direct pistil-specific and pollen-specific activity of the Brassica S locus glycoprotein gene promoter. Plant Cell 5: 855–863 (1993).

    Google Scholar 

  17. Eyal Y, Sagee O, Fluhr R: Dark-induced accumulation of a basic pathogenesis-related (PR-1) transcript and light requirement for its induction by ethylene. Plant Mol Biol 10: 589–599 (1992).

    Google Scholar 

  18. Fluhr R, Sessa G, Sharon A, Ori N, Lotan T: Pathogenesis-related proteins exhibit both pathogen-induced and developmental regulation. In: Advances in Molecular Genetics of Plant-Microbe Interactions, vol 1, pp. 387–394. Kluwer Academic Publishers, dordrecht, Netherlands (1991).

    Google Scholar 

  19. Forde BG: AT-rich elements (ATREs) in the promoter regions of nodulin and other higher plant genes: a novel class of cis-acting regulatory elements? In: Nover L (ed) Plant Promoters and Transcription Factors, pp. 87–103. Springer-Verlag, Berlin/Heidelberg/New York (1994).

    Google Scholar 

  20. Fütterer J, Gordon K, Sanfacon H, Bonneville J-M, Hohn T: Positive and negative control of translation by the leader sequence of cauliflower mosaic virus pregenomic 35S. EMBO J 9: 1697–1707 (1990).

    Google Scholar 

  21. Gallie DR, Sleat DE, Watts JW, Turner PC, Wilson TMA: The 5′-leader sequence of tobacco mosaic virus RNA enhances the expression of foreign gene transcripts in vitro and in vivo. Nucl Acids Res 15: 3257–3273 (1987).

    Google Scholar 

  22. Gasser CS, Robinson-Beers K: Pistil development. Plant Cell 5: 1231–1239 (1993).

    Google Scholar 

  23. Gheysen G, Inzé D, Soetaert P, VanMontagu M, Castresana C: Sequence of a Nicotiana plumbaginifolia β(1,3)-glucanase gene encoding a vacuolar isoform. Nucl Acids Res 18: 6685 (1990).

    Google Scholar 

  24. Gilmartin PM, Sarokin L, Memelink J, Chua NH: Molecular light switches for plant genes. Plant Cell 2: 369–378 (1990).

    Google Scholar 

  25. Hayes TE, Sengupta P, Cochran BH: The human c-fos serum response factor and the yeast factors GRM/PRTF have related DNA-binding specificities. Genes Devel 2: 1713–1722 (1988).

    Google Scholar 

  26. Heslop-Harrison J: Pollen germination and pollen-tube growth. Int Rev Cytol 107: 1–78 (1987).

    Google Scholar 

  27. Hinnebusch AG: Novel mechanisms of translational control in Saccharomyces cerevisiae. Trends Genet 4: 169–174 (1988).

    Google Scholar 

  28. Jansen MAK, Sessa G, Malkin S, Fluhr R: PEPC-mediated carbon fixation in transmitting tract cells reflects style-pollen tube interactions. Plant J 2: 507–515 (1992).

    Google Scholar 

  29. Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).

    Google Scholar 

  30. Joosten MHAJ, Bergmans CJB, Meulenhoff EJS, Cornelissen BJC, deWit PGJM: Purification and serological characterization of three basic 15-kilodalton pathogenesis-related proteins from tomato. Plant Physiol 94: 585–591 (1990).

    Google Scholar 

  31. 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).

    Google Scholar 

  32. Katagiri F, Lam E, Chua NH: Two tobacco DNA-binding proteins with homology to the nuclear factor CREB. Nature 340: 727–730 (1989).

    Google Scholar 

  33. Klarmann GJ, Schauber CA, Preston BD: Templatedirected pausing of DNA synthesis by HIV-1 reverse transcriptase during polymerization of HIV-1 sequences in vitro. J Biol Chem 268: 9793–9802 (1993).

    Google Scholar 

  34. Kozak M: Circumstances and mechanisms of inhibition of translation by secondary structure in eukaryotic mRNAs. Mol Cell Biol 9: 5134–5142 (1989).

    Google Scholar 

  35. Lee H-S, Huang S, Kao T-h: S proteins control rejection of incompatible pollen in Petunia inflata. Nature 367: 560–563 (1994).

    Google Scholar 

  36. Linthorst HJM, Melchers LS, Mayer A, vanRoekel JSC, Cornelissen BJC, Bol JF: Analysis of the gene families encoding acidic and basic β-1,3-glucanases of tobacco. Proc Natl Acad Sci USA 87: 8756–8760 (1990).

    Google Scholar 

  37. Lohmer S, Maddaloni M, Motto M, Salamini F, Thompson RD Transation of the mRNA of the maize transcriptional activator Opaque-2 is inhibited by upstream open reading frames present in the leader sequence. Plant Cell 5: 65–73 (1993).

    Google Scholar 

  38. Lord EM, Sanders LC: Roles for the extracellular matrix in plant development and pollination: a special case of cell movement in plants. Devel Biol 153: 16–28 (1992).

    Google Scholar 

  39. Lotan T, Ori N, Fluhr R: Pathogenesis-related proteins are developmentally regulated in tobacco flowers. Plant Cell 1: 881–887 (1989).

    Google Scholar 

  40. Lutcke HA, Chow KC, Mickel FS, Moss KA, Kern H, Scheele GA: Selection of AUG initiation codons differs in plants and animals. EMBO J 6: 43–48 (1987).

    Google Scholar 

  41. Malehorn DE, Scott KJ, Shah DM: Structure and expression of a barley acidic β-glucanase gene. Plant Mol Biol 22: 347–360 (1993).

    Google Scholar 

  42. Murfett J, Atherton TL, Mou B, Gasser CS, McClure BA. S-RNase expressed in transgenic Nicotiana causes S-allele-specific pollen rejection. Nature 367: 563–566 (1994).

    Google Scholar 

  43. Newbigin E, Anderson MA, Clarke AE: Gametophytic self-incompatibility systems. Plant Cell 5: 1315–1324 (1993).

    Google Scholar 

  44. Ori N, Sessa G, Lotan T, Himmelhoch S, Fluhr R: A major stylar matrix polypeptide (Sp41) is a member of the pathogenesis-related proteins superclass. EMBO J 9: 3429–3426 (1990).

    Google Scholar 

  45. Passmore S, Elble R, Tye BK: A protein involved in minichromosome maintenance in yeast binds a transcriptional enhancer conserved in eukaryotes. Genes Devel 3: 921–935 (1989).

    Google Scholar 

  46. Paulson JC: Glycoproteins: what are the sugar chains for? Trends Biochem Sci 14: 272–276 (1989).

    Google Scholar 

  47. Preissner KT: Structure and biological role of vitronectin. Annu Rev Cell Biol 7: 275–310 (1991).

    Google Scholar 

  48. Sanders LC, Lord EM: Directed movement of latex particles in the gynoecia of three species of flowering plants. Science 243: 1606–1608 (1989).

    Google Scholar 

  49. Sanders LC, Wang C-S, Walling L, Lord EM: A homolog of the substrate adhesion molecule vitronectin occurs in four species of flowering plants. Plant Cell 3: 629–635 (1991).

    Google Scholar 

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

    Google Scholar 

  51. Shiraishi H, Okada K, Shimura Y: Nucleotide sequences recognized by the Agamous MADS domain of Arabidopsis thaliana in vitro. Plant J 4: 385–398 (1993).

    Google Scholar 

  52. Treisman R: The serum response element. Trends Biochem Sci 17: 423–426 (1992).

    Google Scholar 

  53. Vasil IK: Physiology and culture of pollen. Int Rev Cytol 107: 127–174 (1987).

    Google Scholar 

  54. Vögeli-Lange R, Fründt C, Hart CM, Beffa R, Nagy F, MeinsJr F: Evidence for a role of β-1,3-glucanase in dicot seed germination. Plant J 5: 273–278 (1994).

    Google Scholar 

  55. Wang H, Wu H-M, Cheung AY: Development and pollination regulated accumulation and glycosylation of a stylar transmitting tissue-specific proline-rich protein. Plant Cell 5: 1639–1650 (1993).

    Google Scholar 

  56. Wang C-S, Walling LL, Gu YQ, Ware CF, Lord EM: Two classes of proteins and mRNAs in Lilium longiflorum L. identified by human vitronectin probes. Plant Physiol 104: 711–717 (1994).

    Google Scholar 

  57. Worrall D, Hird DL, Hodge R, Paul W, Draper J, Scott R: Premature dissolution of the microsporocyte callose wall causes male sterility in transgenic tobacco. Plant Cell 4: 759–771 (1992).

    Google Scholar 

  58. Zhu J-K, Damsz B, Kononowicz AK, Bressan RA, Hasegawa PM: A higher plant extracellular vitronectin-like adhesion protein is related to the translational elongation factor-1α. Plant Cell 6: 393–404 (1994).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Genetics, Weizmann Institute of Science, P.O.Box 26, 76100, Rehovot, Israel

    Guido Sessa & Robert Fluhr

Authors
  1. Guido Sessa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robert Fluhr
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sessa, G., Fluhr, R. The expression of an abundant transmitting tract-specific endoglucanase (Sp41) is promoter-dependent and not essential for the reproductive physiology of tobacco. Plant Mol Biol 29, 969–982 (1995). https://doi.org/10.1007/BF00014970

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00014970

Key words

Search

Navigation