Abstract
The expression of the starch-synthase gene was studied by in situ RNA hybridization at different stages of pollen development in tobacco (Nicotiana tabacum L.). By means of Northern blot analysis we demonstrated that a potato (Solarium tuberosum L.) gene was homologous to its tobacco counterpart, and then we synthesized 35S-labeled RNA probes from this gene. The probe was seen to bind at the tetrad stage. Unicellular pollen grains showed no starch-synthase mRNA but these transcripts reappeared in the binucleate stages, being abundant in mature pollen grains. Our results show that starch-synthase is expressed in the gametophytic cells. Variations in mRNA accumulation during the different stages indicate a regulation of this gene throughout pollen development.
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Abbreviations
- bp:
-
basepairs
- GBSS:
-
granule-bound starch synthase
References
Angerer, G.L., Angerer, R.C. (1981) Detection of poly A+RNA in sea urchin eggs and embryos by quantitative in situ hybridization. Nucleic Acids Res. 9, 2819–2840
Badenhuizen, N.P. (1969) The biogenesis of starch in higher plants. Appletton-Century-Crofts, New York
Cox, K.H., DeLeon, D.V., Angerer, L.M., Angerer, R.C. (1984) Detection of mRNAs in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev. Biol. 101, 485–502
Frydman, R.B. (1963) Starch synthase of potatoes and waxy maize. Arch. Biochem. Biophys. 102, 242–248
Golberg, R.B. (1988) Plants: novel developmental processes. Science 240, 1460–1467
Hanson, D.D., Hamilton, D.A., Travis, J.L., Bashe, D.M., Mascarenhas, J.P. (1989) Characterization of a pollen-specific cDNA clone from Zea mays and its expression. Plant Cell 1, 173–179
Heberle-Bors, E. (1984) Genotypic control of pollen embryogenesis in Nicotiana tabacum L. Theor. Appl. Genet. 68, 475–479
Heberle-Bors, E., Reinert, J. (1980) Isolated pollen cultures and pollen dimorphism. Naturwissenschaften 67, 311
Jamrich, M.K., Mahon, K.A., Gavis, E.R., Gall, J.G. (1984) Histone RNA in amphibian oocytes visualized by in situ hybridization to methacrylate-embedded tissue sections. EMBO J. 3, 1939–1943
Mascarenhas, J.P. (1975) The biochemistry of Angiosperm pollen development. Bot. Rev. 41, 259–314
McFadden, G.I. (1989) In situ hybridization in plants: from macroscopic to ultrastructural resolution. Cell Biol. Int. Rep. 13, 3–21
Miki-Hirosige, H., Nakamura, S. (1983) Growth and differentiation of amyloplast during male gamete development in Lillium longiflorum. In: Pollen: biology and implications for plant breeding, pp. 141–147, Mulcahy, D.L., Ottaviano, E., eds. Elsevier New York Amsterdam Oxford
Nitsch, J.P., Nitsch, C. (1969) Haploid plants from pollen grains. Science 163, 85–87
Norreel, B. (1970) Etude cytologique de la androgènese chez le Datura inoxia et le Nicotiana tabacum. Bull. Soc. Bot. Fr. 117, 461–478
Raghavan, V. (1986) Embryogenesis in Angiosperms. A developmental and experimental study. Cambridge University Press, Cambridge London New York
Raghavan, V. (1987) Developmental strategies of the angiosperm pollen: a biochemical perspective. Cell Diff. 21, 213–226
Raghavan, V. (1989) mRNAs and a cloned histone gene are differentially expressed during anther and pollen development in rice (Oryza sativa L.) J. Cell Sci. 92, 217–229
Rentrop, M., Knapp, B., Winter, H., Schweizer, J. (1986) Aminoalkylsilane-treated glass slides as support for in situ hybridization of keratin cDNAs to frozen tissue sections under varying fixation and pretreatment conditions. Histochem. J. 18, 271–276
Rodríguez-García, M.I., Garcia, A. (1978) Differentiation of the plastid population during microsporogenesis and the development of the pollen grain in the Liliaceae. Biol. Cell. 33, 63–70
Sangwan, R.S., Sangwan-Norreel, B.S. (1987) Ultrastructural cytology of plastids in pollen grains of certain androgenic and non-androgenic plants. Protoplasma 138, 11–22
Schrauwen, J.A.M., de Groot, P.FM., van Herpen M.M.A., van der Lee, T., Reynen, W.H., Weterings, K.A.P., Wullems, G.J. (1990) Stage-related expression of mRNAs during pollen development in lily and tobacco. Planta 182, 298–304
Shannon, J.C., Garwood, D.L. (1984) Genetics and physiology of starch development. In: Starch: chemistry and technology, pp. 26–86, Whistler R.L., Bemiller J.N., Paschall, E.F., eds. Academic Press, New York
Tsai, J.S., Kuo, C.G. (1980) Enzymatic activities of starch synthesis in potato tubers of different sizes. Physiol. Plant 48, 460–462
Tupy, J., Süss, J., Hrabetova, E., Rihova, L. (1983) Developmental changes in gene expression during pollen differentiation and maturation in Nicotiana tabacum L. Biol. Plant. 25, 231–237
Ursin, V.M., Yamaguchi, J., McCormick, S. (1989) Gametophytic and sporophytic expression of anther-specific genes in developing tomato anthers. Plant Cell 1, 727–736
Visser, R. (1989) Manipulation of the starch composition of Solanum tuberosum L., using Agrobacterium rhizogenes mediated transformation. Ph. D. thesis, Groningen University, The Netherlands
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We are particularly grateful to Dr. Visser (Groningen University, The Netherlands) for generously providing the OBSS clone. We thank Dr. Sánchez-Pina and Dr. Risueño (Centro de Investigaciones Biológicas, Madrid, Spain) for their useful and critical reading of the manuscript. Dr. Olmedilla was the recipient of a C.S.I.C. postdoctoral fellowship during her stay at the Nijmegen University.
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Olmedilla, A., Schrauwen, J.A.M. & Wullems, G.J. Visualization of starch-synthase expression by in situ hybridization during pollen development. Planta 184, 182–186 (1991). https://doi.org/10.1007/BF00197946
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DOI: https://doi.org/10.1007/BF00197946