Abstract
Self-incompatibility in the Solanaceae is controlled by a single multiallelic genetic locus, the S locus. The stylar gene products of the S locus are abundant glycoproteins with ribonuclease activity, secreted in the transmitting tract tissue of the pistil. To investigate the structural and functional integrity and possible phenotypic effects of expression of the S-gene product in the male gametophyte, N. tabacum plants were transformed with a construct containing the genomic S 2 -RNase coding sequence from S. tuberosum under the control of the promoter of the pollen-specific LAT52 gene from tomato. The expression pattern of the S 2 RNase in the male gametophyte at both the protein and RNA level was found to be identical to that already reported for expression of the β-glucuronidase (GUS) gene directed by the LAT52 promoter in transgenic tomato and tobacco. The S 2 -RNase gene fusion led to a tissue-specific and developmentally regulated accumulation of the S 2 polypeptide in pollen of transgenic tobacco plants. The transgenic protein product was of the same size and charge as the potato stylar product, had ribonuclease activity, and was glycosylated. The transgenic plants, however, did not show any morphological variations in their flower organs, and their fertility was not influenced by the accumulation of the S 2 -RNase protein in pollen.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721
De Nettancourt D (1977) Incompatibility in angiosperms. In: Frankel R, Gall GAE, Linskens HF (eds). Monographs on theoretical and applied genetics, vol 3. Springer, Berlin Heidelberg New York
Ditta G, Stanfield S, Corbin D, Helinski DR (1980) Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77:7347–7351
Dodds PN, Bönig I, Du H, Rödin J, Anderson MA, Newbigin E, Clarke AE (1993) S-RNase gene of Nicotiana alata is expressed in developing pollen. Plant Cell 5:1771–1782
Doyle JJ, Doyle JL (1989) Isolation of plant DNA from fresh tissue. BRL Focus 12:13–14
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specifity. Anal Biochem 132:6–13
Gray JE, McClure BA, Bönig I, Anderson MA, Clarke AE (1991) Action of the style product of the self-incompatibility gene of Nicotiana alata (S-RNase) on in vitro-grown pollen tubes. Plant Cell 3:271–283
Haring V, Gray JE, McClure BA, Anderson MA, Clarke AE (1990) Self-incompatibility: a self-recognition system in plants. Science 250:937–941
Hawkes R (1982) Identification of Con A-binding proteins after SDS-gel electrophoresis and protein blotting. Anal Biochem 123:143–146
Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA (1983) A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303:179–180
Horsch RB, Fry JE, Hoffman NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 277:1229–1231
Junghans H, Metzlaff M (1990) A simple and rapid method for the preparation of total plant DNA. BioTechniques 8:176
Kaufmann H (1991) Sequenzvariabilität und Genstruktur des Selbstinkompatibilitäts-Locus in Solanum tuberosum L. PhD thesis, University of Cologne
Kaufmann H, Salamini F, Thompson RD (1991) Sequence variability and gene structure at the self-incompatibility locus of Solanum tuberosum. Mol Gen Genet 226:457–466
Kirch HH, Uhrig H, Lottspeich F, Salamini F, Thompson RD (1989) Characterization of proteins associated with self-incompatibility in Solanum tuberosum. Theor Appl Genet 78:581–588
Koltunow AM, Truettner J, Cox KH, Wallroth M, Goldberg RB (1990) Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2:1201–1224
Kyhse-Andersen J (1984) Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods 10:203–209
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:681–685
Lee H-S, Huang S, Kao T-H (1994) S-proteins control rejection of incompatible pollen in Petunia inflata. Nature 367:560–563
Lewis D (1960) Genetic control of specificity and activity of S-antigen in plants. Proc R Soc Lond B 151:468–477
Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissues. Anal Biochem 163:16–20
McClure BA, Haring V, Ebert PR, Anderson MA, Simpson RJ, Sakiyama F, Clarke AE (1989) Style self-incompatibility gene products of Nicotiana alata are ribonucleases. Nature 342:955–957
McClure BA, Gray JE, Anderson MA, Clarke AE (1990) Self-incompatibility in Nicotiana alata involves degradation of pollen rRNA. Nature 347:757–760
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Murfett J, Cornish EC, Ebert PR, Bönig I, McClure BA, Clarke AE (1992) Expression of a self-incompatibility glycoprotein (S 2 -ribonuclease) from Nicotiana alata in transgenic Nicotiana tabacum. Plant Cell 4:1063–1074
Murfett J, Atherton TL, Mou B, Gasser CS, McClure BA (1994) S-RNase expressed in transgenic Nicotiana causes S-allele-specific pollen rejection. Nature 367:563–566
Sassa H, Hirano H, Ikehashi H (1992) Self-incompatibility-related RNases in styles of Japanese Pear (Pyrus serotina Rehd.). Plant Cell Physiol 33:811–814
Schmidt-Stohn G (1979) Eine Methode zur isoelektrischen Fokussierung von Proteinen in Flachgelen für Serienuntersuchungen in der Genetik. Z Pflanzenzüch 83:81–90
Thompson RD, Kaufmann H, Kirch HH, Salamini F (1991a) Self-incompatibility in flowering plants. In: Harding J, Singh F, Mol JNM (eds) Genetics and breeding of ornamental species. Curr Plant Sci Biotechnol Agri 11: pp 367–385
Thompson RD, Uhrig H, Hermsen JGT, Salamini F, Kaufmann H (1991b) Investigation of a self-incompatible mutation in Solanum tuberosum clones inhibiting S-allele activity in pollen differentially. Mol Gen Genet 226:283–288
Thompson RD, Kirch HH (1992) The S locus of flowering plants: when self-rejection is self-interest. Trends Genet 8:381–387
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Twell D, Wing R, Yamaguchi J, McCormick S (1989) Isolation and expression of an anther-specific gene from tomato. Mol Gen Genet 217:240–245
Twell D, Yamaguchi J, Wing RA, Ushiba J, McCormick S (1990) Pollen-specific gene expression in transgenic plants: coordinate regulation of two different tomato gene promoters during microsporogenesis. Development 109:705–713
Ursin VM, Yamaguchi J, McCormick S (1989) Gametophytic and sporophytic expression of anther-specific genes in developing tomato anthers. Plant Cell 1:727–736
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kirch, HH., Li, YQ., Seul, U. et al. The expression of a potato (Solanum tuberosum) S-RNase gene in Nicotiana tabacum pollen. Sexual Plant Reprod 8, 77–84 (1995). https://doi.org/10.1007/BF00230892
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00230892