Re-examination of the self-incompatibility genotype of apple cultivars containing putative 'new' S-alleles

Recently, the self-incompatibility (S-) genotypes of 56 apple cultivars were examined by protein analysis, which led to the identification by Boskovic and Tobutt of 14 putative 'new' S-alleles, S12 to S25. This paper reports a re-examination of the S-genotypes of some of these cultivars through S-allele 'specific' PCR and sequence analysis. The results obtained by this analysis indicated that the number of S-alleles that are present in apple is probably smaller than the number proposed by Boskovic and Tobutt. The existence of three 'new' S-alleles (S20, S22 and S24) was confirmed. The existence of two other putative 'new' S-alleles (S23 and S25) was, however, contradicted. The coding sequences of the S-alleles that correspond to the SIO and the S25 ribonuclease bands as well as those corresponding to the S22 and the S23 ribonuclease bands were shown to be identical in sequence. Interestingly, the S-allele corresponding to the S22 and the S23 ribonuclease bands shared a high sequence identity (99% identity) with S27, which was previously cloned and sequenced from Baskatong, but which was not included in the analysis conducted by Boskovic and Tobutt. Both S-alleles only differ in point mutations, which are not translated into differences in amino-acid sequence. To our knowledge, this is the first report of two S-alleles that differ at the nucleotide level but still encode for identical S-RNases. The implications of these observations for determining the S-genotypes of plants by PCR analysis or protein analysis are discussed.