Description of a new trans-generic Skb-RNase allele in apple
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Polish apple cvs: ‘Ligol’, ‘Odra’ and ‘Primula’ served for studies of self-incompatibility. Basing on available sequence data, a new set of primers upstream and downstream of the hypervariable (HV) region of apple S-RNases were designed. Using the RT-PCR method, cDNA was amplified on RNA isolated from styles. PCR products were cloned and sequenced. A new trans-generic S-RNase allele, designated as Skb (GenBank accession no. EU443101), was discovered in cvs ‘Odra’ and ‘Primula’. Nucleotide sequence alignment revealed that Skb-RNase shows 98% identity to SaucS19-RNase from Sorbus aucuparia and 97% identity to CmonS17-RNase from Crataegus monogyna. The occurrence of extensive intergeneric hybridization among extant Pyrinae is considered since the deduced amino acid sequence of Skb-RNase from M. × domestica showed higher similarity to CmonS17 from C. monogyna, SaucS19-RNase from S. aucuparia, St from Malus transitoria, S5-RNase and S3-RNase from Pyrus pyrifolia, and S40-RNase from P. ussuriensis than to S-alleles from Malus × domestica and all of them are grouped in the same cluster of phylogenetic tree. In respect to extremely high similarities between aforementioned S-RNases it could be possible that these alleles existed before the separation of Malus, Pyrus, Sorbus and Crataegus genera. Within Malus, the Skb-RNase from M. × domestica and St-RNase from M. transitoria show 100% identity of the HV region at the deduced amino acid level, suggesting that these S-RNases diverged more recently than the other Malus S-RNases. In ‘Ligol’, the agronomically most important cultivar in Poland, the S2 and S9 were identified.