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Genomic characterization of self-incompatibility ribonucleases (S-RNases) in European pear cultivars and development of PCR detection for 20 alleles

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Abstract

Sexual self-incompatibility in European pear (Pyrus communis L.) is controlled by a single locus (S-locus) encoding a polymorphic stylar ribonuclease (S-RNase) that is responsible for the female function in pollen–pistil recognition. In this study, genomic DNA sequences corresponding to five new S-RNase alleles (named S 20 , S 21 , S 22 , S 23 , and S 24 ) and to S m were characterized in European pear cultivars. Re-sequencing S q from ‘General Le Clerc’ showed this S-RNase to encode the same protein as S 12 . Based on these findings, a polymerase chain reaction (PCR)-based method was developed for the molecular typing of cultivars bearing 20 S-RNases (S 1 –S 14 , S m , and S 20 –S 24 ) using consensus and allele-specific primers. Genomic PCR with consensus primers amplified product sizes characteristic of the S-RNases S 1 , S 2 , S 4 , S 10 , S 13 , and S 20 . However, the allele groups S 3 /S 12 , S 6 /S 8 /S 11 /S 22 and S 5 /S 7 /S 9 /S 14 /S m /S 21 /S 23 /S 24 amplified PCR products of similar size. To discriminate between alleles within these groups, primers to specifically amplify each S-RNase were developed. Application of this approach in 19 cultivars with published S-alleles allowed re-evaluation of one of the alleles of ‘Passe Crassane,’ ‘Conference,’ and ‘Condo.’ Finally, this method was used to assign S-genotypes to 37 cultivars. Test crosses confirmed molecular results.

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Abbreviations

HVR:

hyper-variable region

S-RNase:

self-incompatibility ribonuclease

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Acknowledgments

This study was supported by project grants “Consejo Interinstitucional de Ciencia y Tegnología”-AGL2006-13529-C02-02 and “Grupo de Investigación Consolidado” A02-A43. The author is supported by a research contract INIA-CCAA of the “Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria”.

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  1. Unidad de Fruticultura, Centro de Investigación y Tecnología Agroalimentaria de Aragón (CITA), Avenida de Montañana, 930, 50059, Zaragoza, Spain

    Javier Sanzol

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  1. Javier Sanzol
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Correspondence to Javier Sanzol.

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Communicated by E. Dirlewanger

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ESM S1

(PDF 42.24 KB)

ESM S2

Evaluation of allele-specific primers in cultivars harboring Sn (a) and So (b). PCR amplifications using consensus primers (Con) and allele-specific primers for S 3, S 5, S 6, S 7, S 8, S 9, S 11, S 12, S 14, S m, S 21, S 22, S 23 or S 24. Lanes on the gel are: M: Molecular size marker (Invitrogen), 1: a Seedling offspring of the cultivar ‘Forelle’ (S 1 S n) bearing S n (Sanzol, Lozano and Batlle, unpublished result) and b ‘Leonardeta’ (S 1 S o), 2: ‘Precoce Morettini’ (S 1 S 3), 3: ‘Dr. Jules Gujot’ (S 1 S 5), 4: ‘Beurre Giffard’ (S 1 S 6), 5: ‘El dorado’ (S 1 S 7), 6: ‘Starkrimson’ (S 1 S 8), 7: ‘Delbard Premiere’ (S 1 S 9), 8: ‘Wilder’ (S 1 S 11), 9: ‘Pierre Corneille’ (S 1 S 12), 10: ‘Beurré D’Anjou’ (S 1 S 14), 11: ‘Mondsallard’ (S 4 S m), 12: ‘Passe Crassane’ (S 10 S 21), 13: ‘Cure’ (S 3 S 22), 14: ‘Conselleur de la Court’ (S 3 S 23), 15: ‘Green Summer’ (S 22 S 24). (PDF 119.30 KB)

ESM S3

Pollen tube compatibility following cross and self-pollinations in European pear cultivars. (PDF 23.04 KB)

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Sanzol, J. Genomic characterization of self-incompatibility ribonucleases (S-RNases) in European pear cultivars and development of PCR detection for 20 alleles. Tree Genetics & Genomes 5, 393–405 (2009). https://doi.org/10.1007/s11295-008-0194-5

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  • DOI: https://doi.org/10.1007/s11295-008-0194-5

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