Abstract
We report isolation and N-terminal amino acid sequencing of three style glycoproteins, which segregate with three S (self-incompatibility) alleles of Petunia hybrida. The S-glycoproteins were expressed mainly in the upper part of the pistil and showed an increasing concentration during flower development. The glycoproteins were purified by a combination of ConA-Sepharose and cation exchange fast protein liquid chromatography. The amount of S-glycoproteins recovered from style extracts varied from 0.5 to 1.6 μg per style, which was 40–60% of the amount recovered by a simplified analytical method. N-terminal amino acid sequences of S1-, S2- and S3-glycoprotein showed homology within the fifteen amino terminal residues. These amino acid sequences were compared with the previously published sequences of S-glycoproteins from Nicotiana alata and Lycopersicon peruvianum.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Anderson MA, Cornish EC, Mau S-L, Williams EG, Hoggart R, Atkinson A, Bonig I, Grego B, Simpson R, Roche PJ, Haley JD, Niall HD, Tregear GW, Coghlan JP, Crawford RJ, Clarke AE: Cloning of cDNA for a stylar glycoprotein associated with expression of selfincompatibility in Nicotiana alata. Nature 321: 38–44 (1986).
Anderson MA, McFadden GI, Bernatzky R, Atkinson A, Orpin T, Dedman H, Tregear G, Fernley R, Clarke AE: Sequence variability of three alleles of the selfincompatibility gene of Nicotiana alata. Plant Cell 1: 483–491 (1989).
Bradford MM: A rapid sensitive method for the quantification of microgram quantities of protein utilising the principle of protein dye binding. Anal Biochem 72: 248–254 (1976).
Cornish EC, Anderson MA, Clarke AE: Molecular aspects of fertilization in flowering plants. Ann Rev Cell Biol 4: 209–228 (1988).
Cornish EC, Pettitt JM, Bonig I, Clarke AE: Developmentally controlled expression of a gene associated with self-incompatibility in Nicotiana alata. Nature 326: 99–102 (1987).
Herrero M, Dickinson HG: Ultrastructural and physiological differences between buds and mature flowers of Petunia hybrida prior to and following pollination. Planta 148: 138–145 (1980).
Isogai A, Takayama S, Tsukamoto C, Ueda Y, Shiozawa H, Hinata K, Okazaki K, Suzuki A: S-locusspecific glycoproteins associated with self-incompatibility in Brassica campestris. Plant Cell Physiol 28: 1279–1291 (1987).
Jahnen W, Batterham MP, Clarke AE, Moritz RL, Simpson RJ: Identification, isolation and N-terminal sequencing of style glycoproteins associated with selfincompatibility in Nicotiana alata. Plant Cell 1: 493–499 (1989).
Kamboj RK, Jackson JF: Self-incompatibility alleles control a low molecular weight, basic protein in pistils of Petunia hybrida. Theor Appl Genet 71: 815–819 (1986).
Laemmli UK: Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).
Linskens HF: Zur Frage der Abwehrkörper der Inkompatibilitätsreaktion von Petunia. III. Z Bot 48: 16–135 (1960).
Linskens HF: Incompatibility in Petunia. Proc R Soc Lond 188: 299–311 (1975).
Linskens HF, Straub J: A mutant collection of Petunia hybrida. Incomp Newsl 10: 123–131 (1978).
Mau S-L, Raff J, Clarke AE: Isolation and partial characterization of components of Prunus avium styles, including an S-allele-associated antigenic glycoprotein. Planta 156: 505–516 (1982).
Mau S-L, Williams EG, Atkinson A, Anderson MA, Cornish MC, Grego B, Simpson RJ, Kheyr-Pour A, Clarke AE: Style proteins of a wild tomato (Lycopersicon peruvianum) associated with expression of self-incompatibility. Planta 169: 184–191 (1986).
Nasrallah JB, Kao T-H, Goldberg ML, Nasrallah ME: A cDNA clone encoding an S-locus-specific glycoprotein from Brassica oleracea. Nature 318: 263–267 (1985).
Nasrallah JB, Kao T-H, Chen C-H, Goldberg ML, Nasrallah ME: Amino acid sequence of glycoproteins encoded by three alleles of the S-locus of Brassica oleracea. Nature 326: 617–619 (1987).
Nettancourt Dde: Incompatibility in Angiosperms. Springer, Berlin (1977).
Nishio T, Hinata K: Comparative studies on S-glycoproteins purified from different S-genotypes in selfincompatible Brassica species. I. Purification and chemical properties. Genetics 100: 641–647 (1982).
Simpson RJ, Neuberger MR, Liu TY: Complete amino acid analysis of proteins from a single hydrolysate. J Biol Chem 251: 1936 (1976).
Spackman DH, Stein WH, Moore S: Automatic recording apparatus for use in the chromatography of amino acids. Anal Chem 30: 1190–1206 (1958).
Spencer RL, Wold F: A new convenient method for estimation of total cystine-cysteine in proteins. Anal Biochem 32: 185–190 (1969).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Broothaerts, W.J., van Laere, A., Witters, R. et al. Purification and N-terminal sequencing of style glycoproteins associated with self-incompatibility in Petunia hybrida . Plant Mol Biol 14, 93–102 (1990). https://doi.org/10.1007/BF00015658
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00015658