Abstract
A critical stage in pollen development is the dissolution of the four products of meiosis, the tetrads, into free microspores. The tetrads are surrounded by a thick callose wall composed of β-1,3-glucan. At the completion of meiosis, the tetrads are released into the anther locule after hydrolysis of the callose by a β-1,3-glucanase. Using the polymerase chain reaction, we have amplified and subsequently cloned a cDNA corresponding to a β-1,3-glucanase, tobacco (Nicotiana tabacum cv. Samsun) anther glucanase (Tag 1), which is expressed exclusively in anthers from meiosis to the free microspore stage of pollen development. The identity of the clone was determined by DNA and deduced protein sequence similarity to other known β-1,3-glucanases. Several regions strictly conserved among four classes of glucanases are also conserved in the Tag 1 protein. Tag 1 represents a novel class of β-1,3-glucanase based on phylogenetic analysis and RNA expression pattern. Tag 1 RNA was detected in situ only in the tapetum, with maximal expression just prior to tetrad dissolution. Due to its expression pattern and sequence similarity to other β-1,3-glucanases, we believe Tag 1 may be involved in tetrad dissolution.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bedinger P: The remarkable biology of pollen. Plant Cell 4: 879–887 (1992).
Bednarek SY, Raikhel NV: Intracellular trafficking of secretory proteins. Plant Mol Biol 20: 133–150 (1992).
Bhandari N: The microsporangium. In: BM Johri (ed) Embryology of angiosperms, pp. 53–121. Springer-Verlag, Berlin (1984).
Chen L, Fincher GB, Hoj PB: Evolution of polysaccharide hydrolase substrate specificity. J Biol Chem 268: 13318–13326 (1993).
Doyle JJ, Doyle JL: Isolation of plant DNA from fresh tissue. BRL Focus 12: 13–15 (1990).
Echlin P: The role of the tapetum during microsporogenesis of angiosperms. In: J Heslop-Harrison (ed) Pollen Development and Physiology, pp 41–61. Butterworth, London (1971).
Eschrich W, Currier HB: Identification of callose by its diachrome and fluorochrome reactions. Stain Tech 39: 303–307 (1964).
Fincher G, Lock P, Morgan M, Lingelbach K, Wettenhall R, Mercer J, Brandt A, Thomsen K: Primary structure of the (1,3;1,4)-β-D glucohydrolase from barley aleurone. Proc Natl Acad Sci USA 83: 2081–2085 (1986).
Goodspeed TH: The Genus Nicotiana: Origins, Relationships, and Evolution of its Species in Light of Their Distribution, Morphology, and Cytogenetics. Chronica Botanica, Waltham, MA (1954).
Heslop-Harrison J, Mackenzie A: Autoradiography of soluble [2–14C] thymidine derivatives during meiosis and microsporogenesis in Lilium anthers. J Cell Sci 2: 387–400 (1967).
Horner HT, Rogers MA: A comparative light and electron microscopic study of microsporogenesis in male fertile and cytoplasmic male sterile pepper (Capsicum annuum). Can J Bot 52: 435–441 (1974).
Keefe D, Hinz U, Meins F: The effect of ethylene on the cell-type-specific and intracellular localization of β-1,3-glucanase and chitinase in tobacco leaves. Planta 182: 43–51 (1990).
Koltunow AM, Truetner J, Cox KH, Wallroth M, Goldberg RB: Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2: 1201–1224 (1990).
Linthorst H, Melchers L, Mayer A, van Roekel J, Cornelissen B, Bol J: Analysis of gene families encoding acidic and basic β-1,3-glucanases of tobacco. Proc Natl Acad Sci USA 87: 8756–8760 (1990).
Linthorst H: Pathogenesis-related proteins of plants. Crit Rev Plant Sci 10: 123–150 (1991).
MacGregor A, Ballance G: Possible secondary structure in plant and yeast β-glucanases. Biochem J 274: 41–43 (1991).
Mauch F, Hadwiger LA, Boller T: Antifungal hydrolases in pea tissue. I. Purification and characterization of two chitinases and two beta-1,3-glucanases differentially regulated during development and in response to fungal infection. Plant Physiol 87: 325–333 (1988).
Maniatis T, Fritsch E, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Mariani C, De Beuckeleer M, Truettner J, Leemanas J, Goldberg R: Induction of male sterility in plants by a chimeric ribonuclease gene. Nature 347: 737–741 (1990).
Ohno N, Yoshikazu H, Yadomae T: Purification and properties of an exo-(1–3)-β-D-glucanase from the culture filtrate of Mucor hiemalis. Carbohydrate Res 158: 217–226 (1986).
Ohno N, Nono I, Yadomae T: Enzymatic and physiochemical properties of an exo-(1–3)-β-D-glucanase from Rhizoctonia solani. Carbohydrate Res 194: 261–271 (1989).
Okamura 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).
Ori N, Sessa G, Lotan T, Himmelhoch S, Fluhr R: A major stylar polypeptide (sp41) is a member of the pathogenesis-related proteins superclass. EMBO J 9: 3429–3436 (1990).
Payne G, Ward E, Gaffney T, Ahl Goy P, Moyer M, Harper A, Meins FM, Ryals J: Evidence for a third structural class of β-1,3-glucanase in tobacco. Plant Mol Biol 15: 797–808 (1990).
Periasamy K, Swamy BGL: Morphology of the anther tapetum in angiosperms. Curr Sci 35: 427–430 (1966).
Rochester DE, Winter JA, Shah DM: The structure and expression of maize genes encoding the major heat shock protein, hsp70. EMBO J 5: 451–458 (1986).
Shinshi H, Wenzler H, Neuhaus J, Felix G, Meins F: Evidence for N- and C-terminal processing of a plant defense-related enzyme: Primary structure of tobacco prepro-β-1,3-glucanase. Proc Natl Acad Sci USA 85: 5541–5545 (1988).
Smith AG, Hinchee M, Horsch R: Cell and tissue specific expression localized by in situ RNA hybridization in floral tissues. Plant Mol Biol Rep 5: 237–241 (1987).
Smith AG, Gasser CS, Budelier KA, Fraley RT: Identification and characterization of stamen and tapetal specific genes from tomato. Mol Gen Genet 222: 9–16 (1990).
Stieglitz H, Stern H: Regulation of β-1,3-glucanase activity in developing anthers of Lilium. Devel Biol 34: 169–173 (1973).
Stieglitz H: Role of β-1,3-glucanase in postmeiotic microspore release. Dev Biol 57: 87–97 (1977).
Swofford DL: PAUP: Phylogenetic Analysis Using Parsimony, Version 3.0. Computer program distributed by the Illinois Natural History Survey, Champaign, IL (1991).
Takeuchi Y, Yoshikawa M, Takeba G, Kunisuke T, Shibata D, Horino O: Molecular cloning and ethylene induction of mRNA encoding a phytoelexin elicitor-releasing factor β-1,3-glucanase in soybean. Plant Physiol 93: 673–682 (1990).
Van den Bulcke M, Bauw G, Castresana C, Van Montagu M, Vanderchkove J: Characterization of vacuolar and extracellular β-1,3-glucanases of tobacco; evidence for a strictly compartmentalized plant defence system. Proc Natl Acad Sci USA 86: 2673–2677 (1989).
Von Heijne G: A new method for predicting signal sequence cleavage sites. Nucl Acids Res 14: 4683–4690 (1986).
Ward ER, Payne GB, Moyer MB, Williams SC, Dincher SS, Sharkey KC, Beck JJ, Taylor HT, Ahl-Goy P, Meins FM, Ryals JA: Differential regulation of β-1,3-glucanase messenger RNA's in response to pathogen infection. Plant Physiol 96: 390–397 (1991).
Waterkeyn L, Bienfait A: On a possible function of the callosic special wall in Ipomoea purpurea (L) Roth. Grana 10: 13–20 (1970).
Worrall D, Hird D, Hodge R, Wyatt P, Draper J, Scott R: Premature dissolution of the microsporocyte callose wall causes male sterility in transgenic tobacco. Plant Cell 4: 759–771 (1992).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bucciaglia, P.A., Smith, A.G. Cloning and characterization of Tag 1, a tobacco anther β-1,3-glucanase expressed during tetrad dissolution. Plant Mol Biol 24, 903–914 (1994). https://doi.org/10.1007/BF00014444
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00014444