Skip to main content
SpringerLink
Log in

Transgenic ryegrasses (Lolium spp.) with down-regulation of main pollen allergens

  • Published:
Molecular Breeding Aims and scope Submit manuscript

Abstract

Ryegrass pollen (Lolium species) is a widespread source of air-borne allergens and is a major cause of hayfever and seasonal allergic asthma, which affect approximately 25% of the population in cool temperate climates. The main allergens of ryegrass pollen are the proteins Lol p 1 and Lol p 2. These proteins belong to two major classes of grass pollen allergens to which over 90% of pollen-allergic patients are sensitive. The functional role in planta of these pollen allergen proteins remains largely unknown. Here we describe the generation and analysis of transgenic plants with reduced levels of the main ryegrass pollen allergens, Lol p 1 and Lol p 2 in the most important worldwide cultivated ryegrass species, L. perenne and L. multiflorum. These transgenic plants will allow the study of the functional role in planta of these pollen proteins and the determination of potential for development of hypo-allergenic ryegrass cultivars.

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

Abbreviations

AFLP:

amplified fragment length polymorphism

BAC:

bacterial artificial chromosome

DIG:

digoxigenin

GUS:

β-glucuronidase

hph :

hygromycin phosphotransferase

LG:

linkage group

MES:

2-Nmorpholinoethanesulfonicacid

PAGE:

polyacrylamide gel electrophoresis

PBS:

phosphate-buffered saline

PCR:

polymerase chain reaction

PMSF:

phenylmethylsulfonyl fluoride

RFLP:

restriction fragment length polymorphism

SDS:

sodium dodecyl sulphate

References

  • Ansari A.A., Shenbagamurthi P. and Marsh D.G. 1989. Complete primary structure of a Lolium perenne (perennial ryegrass) pollen allergen, Lol p III: comparison with known Lol p I and II sequences. Biochemistry 28: 8665–8670.

    Google Scholar 

  • Altpeter F. and Posselt U.K. 2000. Improved plant regeneration from cell suspensions of commercial cultivars, breeding- and inbred lines of perennial ryegrass (Lolium perenne L.). J. Plant Physiol. 156: 790–796.

    Google Scholar 

  • Altpeter F., Xu J.P. and Ahmed S. 2000. Generation of large numbers of independently transformed fertile perennial ryegrass (Lolium perenne L.). plants of forage- and turf-type cultivars. Mol. Breed. 6: 519–528.

    Google Scholar 

  • Bert P.F., Charmet G., Sourdille P., Hayward M.D. and Balfourier F. 1999. A high density molecular map for ryegrass (Lolium perenne) using AFLP markers. Theor. Appl. Genet. 99: 445–452.

    Google Scholar 

  • Bhalla P.L., Swoboda I. and Singh B.M. 1999. Antisense-mediated silencing of a gene encoding a major ryegrass pollen allergen. Proc. Natl. Acad. Sci. USA 96: 11676–11680.

    Google Scholar 

  • Bieber T. 1996. Fc epsilon R1 on antigen presenting cells. Curr. Opin. Immunol. 8: 773–777.

    Google Scholar 

  • Bilang R., Iida S., Peterhans A., Potrykus I. and Paszkowski J. 1991. The 3’-terminal region of the hygromycin-B-resistance gene is important for its activity in Escherichia coli and Nicotiana tabacum. Gene 100: 247–250.

    Google Scholar 

  • Cosgrove D.J., Bedinger P. and Durachko D.M. 1997. Group I allergens of grass pollen as cell wall-loosening agents. Proc. Natl. Acad. Sci. USA 94: 6559–6564.

    Google Scholar 

  • Esch R.E. and Klapper D.G. 1989. Isolation and characterisation of a major cross-reactive grass group-1 antigenic determinant. Mol. Immunol. 26: 557–561.

    Google Scholar 

  • Esch R.E. and Klapper D.G. 1989. Identification and localization of allergenic determinants on grass group-1 antigens using monoclonal antibodies. J. Immunol. 142: 179–184.

    Google Scholar 

  • Ford S.A. and Baldo B.A. 1986. A re-examination of ryegrass (Lolium perenne) pollen allergens. Int. Arch. Allergy Appl. Immunol. 81: 193–202.

    Google Scholar 

  • Freidhoff L.R., Ehrlich-Kautzky E., Grant J.H., Meyers D.A. and Marsh D.G. 1986. A study of the human immune response to Lolium perenne (rye) pollen and its components, Lol p I and Lol p II (rye I and rye II). I. Prevalence of reactivity to the allergens among skin test, IgE antibody, and IgG antibody data. J. Allergy Clin. Immunol. 78: 1190–1201.

    Google Scholar 

  • Griffith I.J., Smith P.M., Pollock J., Teerakulpisut P., Avjioglu A., Davies S., Hough T., Singh M.B., Simpson R.J., Ward L.D. and Knox B.R. 1991. Cloning and sequencing of Lol p I, the major allergenic protein of ryegrass pollen. FEBS Lett. 279: 210–215.

    Google Scholar 

  • Hamilton D.A., Roy M., Rueda J., Sindhu R.K., Sanford J. and Mascarenhas J.P. 1992. Dissection of a pollen-specific promoter from maize by transient transformation assays. Plant Mol. Biol. 18: 211–218.

    Google Scholar 

  • Helliwell C. and Waterhouse P. 2003. Constructs and methods for high-throughput gene silencing in plants. Methods 30: 289–295.

    Google Scholar 

  • Higo K., Ugawa Y., Iwamoto M. and Higo H. 1998. PLACE: a database of plant cis-acting regulatory DNA elements. Nucleic Acid Res. 26: 358–359.

    Google Scholar 

  • Jauhar P.P. 1993. Cytogenetics of the Festuca-Lolium complex: relevance to breeding. Springer-Verlag, Berlin - New York.

    Google Scholar 

  • Johnson P. and Marsh D.G. 1965. Allergens from common ryegrass pollen (Lolium perenne). The allergenic determinants and carbohydrate moiety. Immunochemistry 3: 101–110.

    Google Scholar 

  • Jones E.S., Mahoney N.L., Hayward M.D., Armstead I.P., Jones J.G., Humphreys M.O., King I.P., Kishida T., Yamada T., Balfourier F., Charmet C. and Forster J.W. 2002a. An enhanced molecular marker-based map of perennial ryegrass (Lolium perenne L.) reveals comparative relationships with other Poaceae species. Genome 45: 282–295.

    Article  CAS  PubMed  Google Scholar 

  • Jones E.S., Dupal M.D., Dumsday J.L., Hughes L.J. and Forster J.W. 2002b. An SSR-based genetic linkage map for perennial ryegrass (Lolium perenne L.). Theor. Appl. Genet. 105: 577–584.

    Google Scholar 

  • Kahn C.R. and Marsh D.G. 1986. Monoclonal antibodies to the major Lolium perenne (rye grass) pollen allergen Lol p 1 (Rye 1). Mol. Immunol. 23: 1281–1288.

    Google Scholar 

  • King T.P., Hoffman D., Lowenstein H., Marsh D.G., Platts-Mills T.A. and Thomas W. 1995. Allergen nomenclature. J. Allergy Clin. Immunol. 96: 5–14.

    Google Scholar 

  • Knox B. and Suphioglu C. 1996. Environmental and molecular biology of pollen allergens. Trends Plant Sci. 1: 156–164.

    Google Scholar 

  • Laffer S., Valenta R., Vrtala S., Susani M., van Ree R., Kraft D., Scheiner O. and Duchene M. 1994. Complementary DNA cloning of the major allergen Phl p I from timothy grass (Phleum pratense); recombinant Phl p I inhibits IgE binding to group I allergens from eight different grass species. J. Allergy Clin. Immunol. 94: 689–698.

    Google Scholar 

  • Li L. and Cosgrove D.J. 2001. Grass group I pollen allergens (β-expansins) lack proteinase activity and do not cause loosening via proteolysis. Eur. J. Biochem. 268: 4217–4226.

    Google Scholar 

  • Muller W.D., Karamfilov T., Bufe A., Fahlbush B., Wolf I. and Jager L. 1996. Group 5 allergens of timothy grass (Phl p 5) bear cross-reacting T cell epitopes with group 1 allergens of ryegrass (Lol p 1). Int. Arch. Allergy Immunol. 109: 352–355.

    Google Scholar 

  • Ong E.K., Griffith I.J., Knox R.B. and Singh M.B. 1993. Cloning of a cDNA encoding a group-V (group-IX) allergen isoform from ryegrass pollen that demonstrates specific antigenic immunoreactivity. Gene 134: 235–240.

    Google Scholar 

  • Perez M., Ishioka Y.G., Walkerm E.L. and Chesnut R. 1990. cDNA cloning and immunological characterization of the ryegrass allergen Lol p I. J. Biol. Chem. 265: 16210–16215.

    Google Scholar 

  • Roberts A.M., Bevan L.J., Flora P.S., Jepson I. and Walker M.R. 1993. Nucleotide sequence of cDNA encoding the group II allergen of cocksfood/orchard grass (Dactylis glomerata), Dac g II. Allergy 48: 615–623.

    Google Scholar 

  • Sambrook J., Fritsch E.F. and Maniatis T. 1989. Molecular Cloning a Laboratory Manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.

    Google Scholar 

  • Sidoli A., Tamborini E., Giuntinis I., Levi S., Volonte G., Paini C., De Lalla C., Siccardi A.G., Barelle F.E., Galliani S. and Arosio P. 1993. Cloning, expression, and immunological characterisation of recombinant Lolium perenne allergen Lol p II. J. Biol. Chem. 268: 21819–21825.

    Google Scholar 

  • Singh M.B., Hough T., Teerakulpisut P., Avjioglu A., Davies S., Smith P.M., Taylor P., Simpson R.J., Ward L.D. and McCluskey J. 1991. Isolation of cDNA encoding a newly identified major allergenic protein of rye-grass pollen: intracellular targeting to the amyloplast. Proc. Natl. Acad. Sci. USA 88: 1384–1388.

    Google Scholar 

  • Singh B.M. and Bhalla L.P. 2003. Hypoallergenic derivatives of major grass pollen allergens for allergy vaccination. Immunol. Cell Biol. 81: 86–91.

    Google Scholar 

  • Smart I.J., Tuddenham W.G. and Knox R.B. 1979. Aerobiology of grass pollen in the city atmosphere of Melbourne: effects of weather parameters and pollen sources. Aust. J. Bot. 27: 333–342.

    Google Scholar 

  • Smith P.M., Ong E.K., Knox R.B. and Singh M.B. 1994. Immunological relationships among Group I and Group V allergens from grass pollen. Mol. Immunol. 31: 491–498.

    Google Scholar 

  • Spangenberg G., Perez Vicente R., Oliveira M.M., Osusky M., Nagel J., Pais M.S. and Potrykus I. 1992. CMS system in Nicotiana: flower development, patterns of mitochondrial DNA and mitochondrial gene expression. Sex. Plant Reprod. 5: 13–26.

    Google Scholar 

  • Spangenberg G., Wang Z.Y., Wu X.L., Nagel J. and Potrykus I. 1995. Transgenic perennial ryegrass (Lolium perenne) plants from microprojectile bombardment of embryogenic suspension cells. Plant Sci. 108: 209–217.

    Google Scholar 

  • Spangenberg G.,Wang Z.Y., Heath R., Kaul V. and Garrett R. 1997. Biotechnology in pasture plant improvement: methods and prospects.. In: Proc XVIII Intl Grassland Congress, Vol. 3. Winnipeg and Saskatoon.

  • Spangenberg G., Wang Z.Y. and Potrykus I. 1998. Biotechnology in forage and turf grass improvement.. In: Frankel R., Grossman M., Linskens H.F., Maliga P. and Riley R. (eds), Monographs on Theoretical and Applied Genetics, Vol. 23. Springer Verlag, Berlin, Heidelberg, Germany, pp. 192.

    Google Scholar 

  • Spangenberg G., Kalla R., Lidgett A., Sawbridge T., Ong E.K. and John U. 2001. Breeding forage plants in the genome era.. In: Spangenberg G. (ed.), Molecular Breeding of Forage Crops. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp.1–39.

    Google Scholar 

  • Stingl G. and Maurer D. 1997. IgE-mediated allergen presentation via Fc epsilon R1 on antigen presenting cells. Int. Arch. Allergy Immunol. 113: 24–29.

    Google Scholar 

  • Tamborini E., Brandazza A., de Lalla C., Musco G., Siccardi A.G., Arosio P. and Sidoli A. 1995. Recombinant allergen Lol p II: expression, purification and characterization. Mol. Immunol. 32: 505–513.

    Google Scholar 

  • Wang Z.Y., Nagel J., Potrykus I. and Spangenberg G. 1993. Plants from cell suspension-derived protoplasts in Lolium species. Plant Sci. 94: 179–193.

    Google Scholar 

  • Wesley S.V., Helliwell C.A., Smith N.A., Wang M., Rouse D.T., Liu Q., Gooding P.S., Singh S.P., Abbott D., Stoutjesdijk P.A., Robinson S.P., Gleave A.P., Green A.G. and Waterhouse P.M. 2001. Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J. 27: 581–590.

    Google Scholar 

  • Xu H., Theerakulpisut P., Goulding N., Suphioglu C., Singh M.B. and Bhalla P.L. 1995. Cloning, expression and immunological characterization of Ory s 1, the major allergen of rice pollen. Gene 164: 255–259.

    Google Scholar 

  • Ye X., Wang Z.Y., Wu X.L., Potrykus I. and Spangenberg G. 1997. Transgenic Italian ryegrass (Lolium multiflorum) plants from microprojectile bombardment of embryogenic suspension cells. Plant Cell Rep. 16: 379–384.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Plant Biotechnology Centre, Primary Industries Research Victoria and Molecular Plant Breeding Cooperative Research Centre, La Trobe University, 3086, Bundoora, Victoria, Australia

    Natasha Petrovska, Rino Donato, Zengyu Wang, Eng-Kok Ong, Elizabeth Jones, John Forster, Michael Emmerling & German Spangenberg

  2. Pioneer Hi-Bred International, 50131, Johnson, Iowa, USA

    Xinli Wu & Elizabeth Jones

  3. Samuel Roberts Noble Foundation, 73401, Ardmore, Oklahoma, USA

    Zengyu Wang

  4. Axxam, San Raffaele Scientific Institute Science Park, 20132, Milan, Italy

    Alessandro Sidoli

  5. Department of Allergy, Immunology and Respiratory Medicine, Alfred Hospital, 3004, Melbourne, Victoria, Australia

    Robyn O’Hehir

Authors
  1. Natasha Petrovska
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xinli Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rino Donato
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zengyu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Eng-Kok Ong
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Elizabeth Jones
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. John Forster
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Michael Emmerling
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Alessandro Sidoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Robyn O’Hehir
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. German Spangenberg
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to German Spangenberg.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Petrovska, N., Wu, X., Donato, R. et al. Transgenic ryegrasses (Lolium spp.) with down-regulation of main pollen allergens. Mol Breeding 14, 489–501 (2005). https://doi.org/10.1007/s11032-005-1011-6

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11032-005-1011-6

Key words

Search

Navigation