Summary
Beta-glucuronidase (GUS) was histochemically analyzed in anthers and pollen of potato, tobacco and tomato. GUS activity was determined in transgenic plants containing a chimaeric GUS gene and in untransformed plants. In anthers of transgenic plants at premeiotic and meiotic stages of sporogenous development, indigogenic precipitation indicative of GUS activity was consistently manifest in cells of the vascular cylinder, the connectivum and the stomium while no activity was found in the tapetal and sporogenous tissues. At similar stages, anther sections of untransformed plants did not show any indigo blue staining. At later stages of microspore and pollen development, anthers of both transgenic and untransformed plants demonstrated consistently high levels of GUS activity in tapetal and sporogenic cells. In anthers of transgenic plants, GUS was also present in the vascular cylinder, the connectivum and the stomium. These results indicate that in anthers of transgenic potato, tobacco and tomato the chimaeric GUS gene product was localized tissue specifically. They also show that an endogenous GUS gene was expressed in a temporal- and spatial-specific manner in the tapetum and pollen of both transformed and untransformed plants.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aoyagi K, Kuhlemeier C, Chua N (1988) The pea rbcS-3A enhancer-like element directs cell-specific expression in transgenic tobacco. Mol Gen Genet 213: 179–185
Aronson NN, de Duve C (1968) Digestive activity of lysosomes: II. The digestion of macromolecular carbohydrates by extracts of rat liver lysosomes. J Biol Chem 243: 4564
Bino RJ, Hille J, Franken J (1987) Kanamycin resistance during in vitro development of pollen from transgenic tomato plants. Plant Cell Rep 6: 333–336
Bino RJ, Franken J, Witsenboer HMA, Hille J, Dons JJM (1988) Effects of Alternaria alternata f.sp. lycopersici toxins on pollen. Theor Appl Genet 76: 204–208
Brewbaker JL, Kwack BH (1963) The essential role of calcium ion in pollen germination and pollen tube growth. Am J Bot 50: 859–865
Dickinson HG, Heslop-Harrison J (1970) The ribosome cycle, nucleoli, and cytoplasmic nucleoloids in the meiocytes of Lilium. Protoplasma 69: 187–200
Ditta G, Stanfield S, Corbin D, Helinski DR (1980) Broad host range DNA cloning system for gram-negative bacteria: construction of gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77: 7347–7351
Gell AC, Bacic A, Clarke AE (1986) Arabinogalactan-proteins of the female sexual tissue of Nicotiana alata. Plant Physiol 82: 885–889
Goldberg RB (1988) Plants: novel developmental processes. Science 240: 1460–1467
Górska-Brylass A (1965) Hydrolases in pollen grains and pollen tubes. Acta Soc Bot Poloniae 34: 589–604
Herrera-Estrella L, Teeri TH, Simpson J (1988) Use of reporter genes to study gene expression in plant cells. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual, vol B1. Kluwer Academic Publishers, Dordrecht Boston London, pp 1–22
Hoekstra FA, Bruinsma J (1979) Protein synthesis of binucleate and trinucleate pollen and its relationship to tube emergence and growth. Planta 146: 559–566
Holt SJ (1958) Indigogenic methods for esterases. In: Danielli IF (ed) General cytochemical methods. Academic Press, New York, pp 375–398
Horsch RB, Fry JE, Hoffmann NL, Wallroth M, Eicholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227: 1229–1231
Horsch RB, Fry JE, Hoffmann NL, Niedermeyer J, Rogers SG, Fraley RT (1988) Leaf disc transformation. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual, vol A5. Kluwer Academic Publishers, Dordrecht Boston London, pp 1–9
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907
Knox RB (1984) Pollen-pistil interactions. In: Linskens HF, Heslop-Harrison J (eds) Cellular interactions. Encycl Plant Physiol 17: 508–608
Koornneef M, Hanhart CJ, Martinelli L (1987) A genetic analysis of cell culture traits in tomato. Theor Appl Genet 74: 633–641
Kroh M, Helsper JPFG (1974) Transmitting tissue and pollen tube growth. In: Linskens HF (ed) Fertilization in higher plants. North-Holland Publishing Company, The Netherlands, pp 167–175
Lojda Z (1971) Indigogenic methods for glycosidases. An improved method for β-glucuronidase. Histochemie 27: 182–192
Lojda Z, Gossrau R, Schiebler TH (1979) Enzyme histochemistry. Springer, Berlin Heidelberg New York, pp 166–172
Maliga P, Breznovitz A, Marton L (1973) Streptomycin resistant plants from callus of resistant tobacco. Nature 244: 29–30
Nagata T, Okada K, Kawazu T, Takebe I (1987) Cauliflower mosaic virus 35S promoter directs S phase specific expression in plant cells. Mol Gen Genet 207: 242–244
Novel G, Novel M (1973) Mutants d'Escherichia coli K 12 affectés pour leur croissance sur méthyl-β-d-glucuronide: Localisation du gène de structure de la β-d-glucuronidase (uidA). Mol Gen Genet 120: 319–335
Ooms G, Hooykaas PJJ, Van Veen RJM, Van Beelen P, Regensburg-Tuiunk AJG, Schilperoort RA (1982) Octopine Ti plasmid deletion mutants of Agrobacterium tumefaciens with emphasis on the right side of the T-region. Plasmid 7: 15–25
Pedersen S, Simonsen V, Loeschke V (1987) Overlap of gametophytic and sporophytic gene expression in barley. Theor Appl Genet 75: 200–206
Sari Gorla M, Frova C, Binelli G, Ottaviano E (1986) The extent of gametophytic-sporophytic gene expression in maize. Theor Appl Genet 72: 42–47
Searcy KB, Mulcahy DL (1985) The parallel expression of metal tolerance in pollen and sporophytes in Silene dioica, S. alba, and Mimulus guttatus. Theor Appl Genet 69: 597–602
Shillito R, Saul M, Paszkowski J, Potrykus I (1985) High efficiency direct gene transfer to plants. Biotechnology 3: 1099–1103
Singh MB, Knox RB (1984) Quantitative cytochemistry of β-galactosidase in normal and enzyme deficient (gal) pollen of Brassica campestris: application of the indigogenic method. Histochem J 16: 1273–1296
Singh MB, O'Neill PM, Knox RB (1985) Initiation of postmeiotic β-galactosidase synthesis during microsporogenesis in oilseed rape. Plant Physiol 77: 225–228
Sood PP (1980) Histoenzymological compartmentation of β-glucuronidase in the germinating pollen grains of Portulaca grandiflora. Biol Plant (Praha) 22: 124–127
Stiekema WJ, Heidekamp F, Louwerse JD, Verhoeven HA, Dijkhuis P (1988) Introduction of foreign genes into potato Bintje and Désirée using an Agrobacterium tumefaciens binary vector. Plant Cell Rep 7: 47–50
Stinson JR, Mascarenhas JP (1985) Onset of alcohol dehydrogenase synthesis during microsporogenesis in maize. Plant Physiol 77: 222–224
Tanksley SD, Zamir D, Rick CM (1981) Evidence for extensive overlap of sporophytic and gametophytic gene expression in Lycopersicon esculentum. Science 213: 453–455
Töpfer R, Pröls M, Schell J, Steinbiß H (1988) Transient gene expression in tobacco protoplasts: Comparison of the reporter gene systems for CAT, NPTII, and GUS. Plant Cell Rep 7: 225–228
Vasil IK (1967) Physiology and cytology of anther development. Biol Rev 42: 327–373
Vithanage HIMV, Knox RB (1976) Pollen wall proteins: quantitative cytochemistry of the origins of intine and exine enzymes in Brassica oleracea. J Cell Sci 21: 423–435
Vithanage HIMV, Knox RB (1979) Pollen development and quantitative cytochemistry of exine and intine enzymes in sunflower, Helianthus annuus. Ann Bot 44: 95–106
Willing RP, Bashe D, Mascarenhas JP (1988) An analysis of the quantity and diversity of messenger RNAs from pollen and shoots of Zea mays. Theor Appl Genet 75: 751–775
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Plegt, L., Bino, R.J. β-Glucuronidase activity during development of the male gametophyte from transgenic and non-transgenic plants. Mol Gen Genet 216, 321–327 (1989). https://doi.org/10.1007/BF00334371
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00334371