Summary
Bet v 1 proteins, a group of isoforms that constitute the major pollen allergen of the white birch (Betula verrucosa), are encoded by a complex multigene family which also includes a subset of genes transcriptionally activated in cell suspension cultures in the presence of microbial pathogens. We have investigated the temporal and spatial expression patterns ofBet v 1 genes during development of male birch inflorescences. Western and Northern analysis indicated that the expression of Bet v 1 isoallergens is regulated at the transcriptional level and activated only during the late phases of anther maturation, so that high levels of these proteins accumulate in mature pollen. In situ hybridization experiments, carried out with sections of male inflorescences isolated at defined developmental stages, showed thatBet v 1 genes are specifically expressed in late bicellular and mature pollen, but no transcripts were detected in sporophytic anther tissues of any developmental stage. These data suggest a possible role of Bet v 1 proteins during pollen germination on the stigma, protecting the female reproductive tissues from infection by pathogens.
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Abbreviations
- BCIP:
-
5-bromo-4-chlor-3-indolylphosphate
- BSA:
-
bovine serum albumin
- DIECA:
-
sodium diethyldithiocarbamate
- NBT:
-
4-nitro blue tetrazolium chloride
- PMSF:
-
phenylmethylsulfonyl fluoride
References
Bradford M (1976) 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
Breiteneder H, Pettenburger K, Bito A, Valenta R, Kraft D, Rumpold H, Scheiner O, Breitenbach M (1989) The gene coding for the major birch pollen allergenBet v 1 is highly homologous to a pea disease resistance response gene. EMBO J 8: 1935–1938
Dickinson H (1994) Simply a social disease? Nature 367: 517–518
Ferreira FD, Hoffmann-Sommergruber K, Breiteneder H, Pettenburger K, Ebner C, Sommergruber W, Steiner R, Bohle B, Sperr WR, Valent P, Kungl AJ, Breitenbach M, Kraft D, Scheiner O (1993) Purification and characterization of recombinantBet v 1, the major birch pollen allergen. Immunological equivalence to naturalBet v 1. J Biol Chem 268: 19574–19580
Fristensky B, Horovitz D, Hadwiger LA (1988) cDNA sequences for pea disease resistance response genes. Plant Mol Biol 11: 713–715
Grote M (1992) Ultrastructural morphology of an allergen in birch pollen after aqueous, anhydrous-liquid, and vapor fixation technique. J Microsc Res Tech 21: 242–248
Ipsen H, Loewenstein H (1983) Isolation and immunochemical characterization of the major allergen to birch pollen (Betula verrucosa). J Allergy Clin Immunol 72: 150–195
Jarolim E, Tejkl M, Rohac M, Schlerka G, Scheiner O, Kraft D, Breitenbach M, Rumpold H (1989) Monoclonal antibodies against birch pollen allergens: characterisation by immunoblotting and use for single step affinity purification of the major allergenBet v 1. Int Arch Allergy Appl Immunol 90: 54–60
Knox RB, Heslop-Harrison J (1971) Pollen-wall proteins: localization of antigenic and allergenic proteins in the pollen grain wallsof Ambrosia spp. (ragweeds). Cytobios 4: 49–54
— —, Reed C (1970) Localization of antigens associated with the pollen grain wall by immunofluorescence. Nature 225: 1066–1068
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, New York
Mascarenhas JP (1990) Gene activity during pollen development. Annu Rev Plant Physiol Plant Mol Biol 41: 317–338
Matton DP, Brisson N (1989) Cloning, expression, and sequence conservation of pathogenesis-related gene transcripts of potato. Mol Plant Microbe Interact 2: 325–331
Melton DA, Krieg PA, Rebagliatti MR, Maniatis T, Zinn K, Green MR (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res 12: 7035–7056
Miki-Hirosige H, Nakamura S, Yaseuda H, Shida T, Takahashi Y (1994) Immunocytochemical localization of the allergenic proteins in the pollen ofCryptomeria japonica. Sex Plant Reprod 7: 95–100
Simon EW (1974) Phospholipid and plant membrane permeability. New Phytol 73: 377–420
Somssich IE, Schmelzer E, Kawalleck P, Hahlbrock K (1988) Gene structure and in situ transcript localization of pathogenesis-related protein 1 in parsley. Mol Gen Genet 213: 93–98
Staff IA, Taylor PE, Smith P, Singh MB, Knox RB (1990) Cellular localization of water soluble, allergenic proteins in rye-grass (Lolium perenne) pollen using monoclonal and specific IgE antibodies with immunogold probes. Histochemistry 22: 276–290
Swoboda I, Scheiner O, Kraft D, Breitenbach M, Heberle-Bors E, Vicente O (1994) A birch gene family encoding pollen allergens and pathogenesis-related proteins. Biochim Biophys Acta 1219: 457–464
—, Jilek A, Ferreira F, Engel E, Hoffmann-Sommergruber K, Scheiner O, Kraft D, Breiteneder H, Pittenauer E, Schmid E, Vicente O, Heberle-Bors E, Ahorn H, Breitenbach M (1995) Isoforms of Bet v 1, the major birch pollen allergen, analyzed by liquid chromatography, mass spectrometry, and cDNA cloning. J Biol Chem 270: 2607–2613
Walter MH, Liu J-W, Grand C, Lamb CJ, Hess D (1990) Bean pathogenesis-related (PR) proteins deduced from elicitor-induced transcripts are members of a ubiquitous new class of conserved PR proteins including pollen allergens. Mol Gen Genet 222: 353–360
Yeh K-W, Juang R-H, Su J-C (1991) A rapid and efficient method for RNA isolation from plants with high carbohydrate content. Focus 13: 102–103
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Swoboda, I., Dang, T.C.H., Heberle-Bors, E. et al. Expression of Bet v 1, the major birch pollen allergen, during anther development. Protoplasma 187, 103–110 (1995). https://doi.org/10.1007/BF01280237
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DOI: https://doi.org/10.1007/BF01280237