Plant Molecular Biology

, Volume 20, Issue 3, pp 493–502 | Cite as

Isolation and characterization of a tobacco gene with homology to pectate lyase which is specifically expressed during microsporogenesis

  • H. J. Rogers
  • A. Harvey
  • D. M. Lonsdale
Research Article

Abstract

A genomic clone has been isolated which contains an open reading frame of 1191 bp interrupted by two small introns. The ORF has been sequenced and the transcriptional start determined. The predicted amino acid sequence shows homology to the deduced amino acid sequences of two pollen-specific pectate lyase genes identified in tomato. The genomic clone was isolated using a partial cDNA clone, TP10, which had been isolated from a Nicotiana tabacum pollen cDNA library by means of differential screening. TP10 has been fully sequenced and contains an open reading frame of 792 bp which shows 96% homology to the ORF in the genomic clone. The transcript corresponding to TP10 is maximally expressed late in pollen development, and has not been detected in vegetative tissues.

Key words

gene expression microsporogenesis Nicotiana tabacum pectate lyase pollen 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Albani D, Altosaar I, Arnison PG, Fabijanski F: A gene showing sequence similarity to pectin esterase is specifically expressed in developing pollen of Brassica napus. Sequences in its 5′ flanking region are conserved in other pollen-specific promoters. Plant Mol Biol 16: 501–513 (1991).Google Scholar
  2. 2.
    Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JD, Smith JA, Struhl K (eds) Current Protocols in Molecular Biology, Wiley, New York (1990).Google Scholar
  3. 3.
    Baulcombe DC, Buffard D: Gibberellic-acid-regulated expression of alpha amylase and six other genes in wheat aleurone layers. Planta 157: 493–500 (1983).Google Scholar
  4. 4.
    Brown SM, Crouch ML: Characterisation of a gene family abundantly expressed in Oenothera organensis pollen that shows sequence similarity to polygalacturonase. Plant Cell 2: 263–274 (1990).Google Scholar
  5. 5.
    Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programmes for the VAX. Nucl Acids Res 12: 387–395 (1984).Google Scholar
  6. 6.
    Feinberg AP, Vogelstein B: A technique for radiolabelling DNA restriction fragments to high specific activity. Anal Biochem 132: 6–13 (1983).Google Scholar
  7. 7.
    Grierson D, Tucker GA, Keen J, Ray J, Bird CR, Schuch W: Sequencing and identification of a cDNA clone for tomato polygalacturonase. Nucl Acids Res 14: 8595–8603 (1986).Google Scholar
  8. 8.
    Gubler U, Hoffman BJ: A simple and very efficient method of generating cDNA libraries. Gene 25: 263–269 (1983).Google Scholar
  9. 9.
    Guerrero FD, Crossland L, Smutzer GS, Hamilton DA, Mascarenhas JP: Promoter sequences from a maize pollen-specific gene direct tissue-specific transcription in tobacco. Mol Gen Genet 224: 161–168 (1990).Google Scholar
  10. 10.
    Higgins DG, Sharp PM: CLUSTAL: a package for performing multiple sequence alignments on a microcomputer. Gene 73: 237–244 (1988).Google Scholar
  11. 11.
    Hinton JCD, Sidebotham JM, Gill DR, Salmond GPC: Extracellular and periplasmic isoenzymes of pectate lyase from Erwinia carotovora subspecies carotovora belong to different gene families. Mol Microbiol 3: 1785–1795 (1989).Google Scholar
  12. 12.
    Huyn TV, Young RA, Davis RW: Constructing and screening cDNA libraries in lambda gt10 and gt11. In: Glover DM (ed) DNA Cloning Techniques — A Practical Approach, vol. 1, pp. 49–78. IRL Press, Oxford (1985).Google Scholar
  13. 13.
    Joshi CP: An inspection of the domain between putative TATA box and translational start site in 79 plant genes. Nucl Acids Res 15: 6648–6653 (1987).Google Scholar
  14. 14.
    Kaiser K, Murray NE: The use of Phage Lambda replacement vectors in the construction of representative genomic DNA libraries. In: Glover DM (ed) DNA Cloning Techniques — A Practical Approach, vol. 1, pp. 1–47. IRL Press, Oxford (1985).Google Scholar
  15. 15.
    Keen NT, Tamaki S: Structure of two pectate lyase genes from Erwinia chrysanthemi EC16 and their high-level expression in Escherichia coli. J Bact 168: 595–606 (1986).Google Scholar
  16. 16.
    Koltunow AM, Truetter J, Cox KH, Wallroth M, Goldberg RB: Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2: 1201–1224 (1991).Google Scholar
  17. 17.
    Kyo M, Harada H: Control of the developmental pathway of tobacco pollen in vitro. Planta 168: 427–432 (1986).Google Scholar
  18. 18.
    Lazarus CM, Baulcombe DC, Martienssen RA: Alpha-amylase genes of wheat are two multigene families which are differentially expressed. Plant Mol Biol 5: 8–24 (1985).Google Scholar
  19. 19.
    Lei S-P, Lin H-C, Wang S-S, Callaway J, Wilcox G: Characterisation of the Erwinia carotovora pelB gene and its product pectate lyase. J Bact 169: 4379–4383 (1987).Google Scholar
  20. 20.
    Lei S-P, Lin H-C, Wand S-S, Wilcox G: Characterisation of the Erwinia carotovora pelA gene and its product pectate lyase A. Gene 62: 159–164 (1988).Google Scholar
  21. 21.
    Mascarenhas JP: Pollen tube growth and RNA synthesis by tube and generative nuclei of Tradescantia. Am J Bot 52: 617 (1965).Google Scholar
  22. 22.
    Mascarenhas JP: Pollen tube growth and ribonucleic acid synthesis by vegetative and generative nuclei of Tradescantia. Am J Bot 53: 563–569 (1966).Google Scholar
  23. 23.
    Mascarenhas JP: Gene activity during pollen development. Annu Rev Plant Physiol Plant Mol Biol 41: 317–338 (1990).Google Scholar
  24. 24.
    McCormick S: Molecular analysis of male gametogenesis in plants. Trends Genet 7: 298–303 (1991).Google Scholar
  25. 25.
    Murphy G, Kavanagh TA: Speeding up the sequencing of double-stranded DNA. Nucl Acids Res 16: 5198 (1988).Google Scholar
  26. 26.
    Neale AD, Wahleithner JA, Lund M, Bonnett HT, Kelly A, Meeks-Wagner DR, Peacock WJ, Dennis ES: Chitinase, β-1,3-glucanase, osmotin, and extensin are expressed in tobacco explants during flower formation. Plant Cell 2: 673–684 (1990).Google Scholar
  27. 27.
    Niogret MF, Dubald M, Mandaron P, Mache R: Characterisation of pollen polygalacturonase encoded by several cDNA clones in maize. Plant Mol Biol 17: 1155–1164 (1991).Google Scholar
  28. 28.
    Okamuro JK, Goldberg RB: Tobacco single copy DNA is highly homologous to sequences present in the genomes of its diploid progenitors. Mol Gen Genet 198: 290–298 (1985).Google Scholar
  29. 29.
    Pressey R, Reger BJ: Polygalacturonase in pollen from corn and other grasses. Plant Sci 59: 57–62 (1989).Google Scholar
  30. 30.
    Rogers HJ, Lonsdale DM: Genetic manipulation of male sterility for the production of hybrid seed. Plant Growth Regul 11: 21–26 (1992).Google Scholar
  31. 31.
    St. John TP, Davis RW: Isolation of galactose-inducible DNA sequences from Saccharomyces by differential plaque filter hybridization. Cell 16: 443–552 (1979).Google Scholar
  32. 32.
    Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1989).Google Scholar
  33. 33.
    Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).Google Scholar
  34. 34.
    Tamaki SJ, Gold S, Robeson M, Manulis S, Kenn NT: Structure and organisation of the pel genes from Erwinia chrysanthemi EC16. J Bact 170: 3468–3478 (1988).Google Scholar
  35. 35.
    Taniai M, Ando S, Usui M, Kurimoto M, Sakaguchi M, Inouye S, Matuhasi T: N-terminal amino acid sequence of a major allergen of Japanese cedar pollen (Cry jI). FEBS Lett 239: 329–332 (1988).Google Scholar
  36. 36.
    Twell D, Yamaguchi J, Wing RA, Ushiba J, McCormick S: Promoter analysis of genes that are coordinately expressed during pollen development reveals pollen-specific enhancer sequences and shared regulatory elements. Genes Devel 5: 496–507 (1991).Google Scholar
  37. 37.
    van Tunen AJ, Mur LA, Brouns GS, Reinstra J-D, Koes RE, Mol JNM: Pollen- and anther-specific promoters from petinia: Tandem promoter regulation of the chiA gene. Plant Cell 2: 393–401 (1990).Google Scholar
  38. 38.
    von Heijne G: A new method for predicting signal sequence cleavage sites. Nucl Acids Res 14: 4683–4690 (1983).Google Scholar
  39. 39.
    Wing RA, Yamaguchi J, Larabell SK, Ursin VM, McCormick S: Molecular and genetic characterisation of two pollen expressed genes that have sequence similarity to pectate lyases of the plant pathogen Erwinia. Plant Mol Biol 14: 17–28 (1989).Google Scholar

Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • H. J. Rogers
    • 1
  • A. Harvey
    • 1
  • D. M. Lonsdale
    • 1
  1. 1.The Cambridge Laboratory, AFRC Institute of Plant Science ResearchJohn Innes CentreNorwichUK

Personalised recommendations