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Isolation of S-locus F-box alleles in Prunus avium and their application in a novel method to determine self-incompatibility genotype

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Abstract

This study characterises a series of 12 S-locus haplotype-specific F-box protein genes (SFB) in cherry (Prunus avium) that are likely candidates for the pollen component of gametophytic self-incompatibility in this species. Primers were designed to amplify 12 SFB alleles, including the introns present in the 5′ untranslated region; sequences representing the S-alleles S 1 , S 2 , S 3 , S 4 , S 4 ′, S 5 , S 6 , S 7 , S 10 , S 12 , S 13 and S 16 were cloned and characterized. [The nucleotide sequences reported in this paper have been submitted to the EMBL/GenBank database under the following accession numbers: PaSFB 1 (AY805048), PaSFB 2 (AY805049), PaSFB 3 (AY805057), PaSFB 4 (AY649872), PaSFB 4 ′ (AY649873), PaSFB 5 (AY805050), PaSFB 6 (AY805051), PaSFB 7 (AY805052), PaSFB 10 (AY805053), PaSFB 12 (AY805054), PaSFB 13 (AY805055), PaSFB 16 (AY805056).] Though the coding regions of six of these alleles have been reported previously, the intron sequence has previously been reported only for S 6 . Analysis of the introns revealed sequence and length polymorphisms. A novel, PCR-based method to genotype cultivars and wild accessions was developed which combines fluorescently labelled primers amplifying the intron of SFB with similar primers for the first intron of S-RNase alleles. Intron length polymorphisms were then ascertained using a semi-automated sequencer. The convenience and reliability of this method for the determination of the self-incompatibility (SI) genotype was demonstrated both in sweet cherry cultivars representing alleles S 1 to S 16 and in individuals from a wild population encompassing S-alleles S 17 to S 22 . This method will greatly expedite SI characterisation in sweet cherry and also facilitate large-scale studies of self-incompatibility in wild cherry and other Prunus populations.

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Abbreviations

GSI:

Gametophytic self-incompatibility

SI:

Self-incompatibilty

SFB :

S-locus F-box

SLFL :

Non-haplotype-specific S-locus F-box

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Acknowledgements

This research was supported by funding from Defra and the Forestry Commission (Grant no.WD0502). The authors would also like to thank Bart De Cuyper, Dr. Tineke Sonneveld and Dr. Radovan Bošković for provision of primers and P. avium material and for many helpful discussions.

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Correspondence to S. P. Vaughan.

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Communicated by H. Nybom

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Vaughan, S.P., Russell, K., Sargent, D.J. et al. Isolation of S-locus F-box alleles in Prunus avium and their application in a novel method to determine self-incompatibility genotype. Theor Appl Genet 112, 856–866 (2006). https://doi.org/10.1007/s00122-005-0187-9

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  • DOI: https://doi.org/10.1007/s00122-005-0187-9

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