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A molecular method for S-allele identification in apple based on allele-specific PCR

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Abstract

cDNA sequences corresponding to two self-incompatibility alleles (S-alleles) of the apple cv ‘Golden Delicious’ have previously been described, and now we report the identification of three additional S-allele cDNAs of apple, one of which was isolated from a pistil cDNA library of cv ‘Idared’ and two of which were obtained by reverse transcription-PCR (RT-PCR) on pistil RNA of cv ‘Queen's Cox’. A comparison of the deduced amino acid sequences of these five S-allele cDNAs revealed an average homology of 69%. Based on the nucleotide sequences of these S-allele cDNAs, we developed a molecular technique for the diagnostic identification of the five different S-alleles in apple cultivars. The method used consists of allele-specific PCR amplification of genomic DNA followed by digestion of the amplification product with an allele-specific restriction endonuclease. Analysis of a number of apple cultivars with known S-phenotype consistently showed coincidence of phenotypic and direct molecular data of the S-allele constitution of the cultivars. It is concluded that the S-allele identification approach reported here provides a rapid and useful method to determine the S-genotype of apple cultivars.

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References

  • Aldrich J, Cullis CA (1993) RAPD analysis in flax: optimisation of yield and reproduction using Klen TaqI DNA polymerase, Chelex 100, and gel purification of genomic DNA. Plant Mol Biol Rep 11:128–141

    Google Scholar 

  • Brace J, King GJ, Ockendon DJ (1994) A molecular approach to the identification of S-alleles in Brassica oleracea. Sex Plant Reprod 7:203–208

    Google Scholar 

  • Broothaerts W, Vanvinckenroye P, Decock B, Van Damme J, Vendrig JC (1991) Petunia hybrida S-proteins:ribonuclease acitivity and the role of their glycan side chains in self-incompatibility. Sex Plant Reprod 4:258–266

    Google Scholar 

  • Broothaerts W, Janssens GA, Proost P, Broekaert WF (1995) cDNA cloning and molecular analysis of two self-incompatibility alleles from apple. Plant Mol Biol 27, 499–511

    Google Scholar 

  • Clarke AE, Newbegin ED (1993) Molecular aspects of self-incompatibility in flowering plants. Annu Rev Genet 27:257–279

    Google Scholar 

  • Dayhoff MO, Schwarts RM, Orcutt BC (1979) A model of evolutionary change in proteins. In: Dayhoff MO (ed) Atlas of protein sequence and structure, vol 5 (suppl 3). National Biomedical Research Foundation, Washington D.C., pp. 345–352

    Google Scholar 

  • de Nettancourt D (1977) Incompatibility in angiosperms. In: Frankel R, Gall GAE, Linskens HF (eds) Monographs in theoretical and applied genetics, vol 3. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Green PJ (1994) The ribonucleases of higher plants. Annu Rev Plant Physiol Plant Mol Biol 45:421–445

    Google Scholar 

  • Ioerger TR, Clark AG, Kao T-h (1990) Polymorphism at the self-incompatibility locus in Solanaceae predates speciation. Proc Natl Acad Sci USA 87:9732–9735

    Google Scholar 

  • Ioerger TR, Gohlke JR, Xu B, Kao T-h (1991) Primary structural features of the self-incompatibility protein in Solanaceae. Sex Plant Reprod 4:81–87

    Google Scholar 

  • 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

    Google Scholar 

  • Kawata Y, Sakiyama F, Tamaoki H (1988) Amino acid sequence of ribonuclease T2 from Aspergillus oryzae. Eur J Biochem 176:683–697

    Google Scholar 

  • Kawata Y, Sakiyama F, Hayashi F, Kyogoku Y (1990) Identification of two essential histidine residues of ribonuclease T2 from Aspergillus oryzae. Eur J Biochem 187:255–262

    Google Scholar 

  • Kobel F, Steinegger P, Anliker J (1939) Weitere Untersuchungen über die Befruchtungsverhältnisse der Apfel-und Birnensorten. Landw Jb Schweiz pp 160–191

  • Lee H-s, Singh A, Kao T-h (1992) RNase X2, a pistil-specific ribonuclease from Petunia inflata, shares sequence similarity with solanaceous S-proteins. Plant Mol Biol 20:1131–1141

    Google Scholar 

  • Lee H-s, Huang S, Kao T-h (1994) S-protein rejection of incompatible pollen in Petunia inflata. Nature 367:560–563

    Google Scholar 

  • Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissues. Anal Biochem 163:16–20

    CAS  PubMed  Google Scholar 

  • Manganaris AG, Alston FH (1987) Inheritance and linkage relationships of glutamate oxaloacetate transaminae isoenzymes in apple. I. The GOT-1, a marker for the S-incompatibility locus. Theor Appl Genet 74:154–161

    Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    CAS  PubMed  Google Scholar 

  • Sassa H, Hirano H, Ikehashi H (1992) Self-incompatibility-related RNases in styles of Japanese pear (Pyrus serotina Redh.) Plant Cell Physiol 33:811–814

    Google Scholar 

  • Sassa H, Hirano H, Ikehashi H (1993) Identification and characterization of stylar glycoproteins associated with self-incompatibility genes of Japanese pear, Pyrus serotina Redh. Mol Gen Genet 241:17–25

    Google Scholar 

  • Sassa H, Mase N, Hirano H, Ikehashi H (1994) Identification of selfincompatibility-related glycoproteins in styles of apple (Malus x domestica). Theor Appl Genet 89:201–205

    Google Scholar 

  • Singh A, Ai Y, Kao T-h (1991) Characterization of ribonuclease activity of three S-allele-associated proteins of Petunia inflata. Plant Physiol 96:61–68

    Google Scholar 

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Communicated by H. F. Linskens

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Janssens, G.A., Goderis, I.J., Broekaert, W.F. et al. A molecular method for S-allele identification in apple based on allele-specific PCR. Theoret. Appl. Genetics 91, 691–698 (1995). https://doi.org/10.1007/BF00223298

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  • DOI: https://doi.org/10.1007/BF00223298

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