No abstract

The molecular bases of the gametophytic self-incompatibility (GSI) system of species of the subtribe Pyrinae (Rosaceae), such as apple and pear, have been widely studied in the last two decades. The characterization of S-locus genes and of the mechanisms underlying pollen acceptance or rejection have been topics of major interest. Besides the single pistil-side S determinant, the S-RNase, multiple related S-locus F-box genes seem to be involved in the determination of pollen S specificity. Here, we collect and review the state of the art of GSI in the Pyrinae. We emphasize recent genomic data that have contributed to unveiling the S-locus structure of the Pyrinae, and discuss their consistency with the models of self-recognition that have been proposed for Prunus and the Solanaceae. Experimental data suggest that the mechanism controlling pollen-pistil recognition specificity of the Pyrinae might fit well with the collaborative 'non-self' recognition system proposed for Petunia (Solanaceae), whereas it presents relevant differences with the mechanism exhibited by the species of the closely related genus Prunus, which uses a single evolutionarily divergent F-box gene as the pollen S determinant. The possible involvement of multiple pollen S genes in the GSI system of Pyrinae, still awaiting experimental confirmation, opens up new perspectives to our understanding of the evolution of S haplotypes, and of the evolution of S-RNase-based GSI within the Rosaceae family. Whereas S-locus genes encode the players determining self-recognition, pollen rejection in the Pyrinae seems to involve a complex cascade of downstream cellular events with significant similarities to programmed cell death.

Pollen–pistil incompatibility in european pear (Pyrus communis L.) compromises adequate orchard pollination and fruit set and restricts cross-fertility between cultivars suitable as parents in breeding programs. Genetic control is simple, with a single locus expressed gametophytically in pollen controlling the rejection of the pollen tube in the style. Semicompatible pollination arises when only one allele of a pollen parent matches the pistil. Semicompatible test-crosses using partially S-genotyped european pear cultivars allowed the discrimination of 14 S-alleles (S1 to S14) at the phenotypic level and the assignment of 33 cultivars to 13 incompatibility groups. Partial genomic sequences of the S-RNase gene, spanning between the C1 and C5 conserved regions, were obtained for each new S-allele identified (S6 to S14). These sequences and those reported previously for the S1 to S5 RNases allowed a set of consensus primers amplifying all 14 S-RNase alleles to be designed. Allele-specific PCR allowed discrimination between those S-RNases giving amplification products of similar size with consensus primers. These two approaches provided a method for the molecular identification of all 14 S-alleles in european pear. With this methodology, we demonstrate that the S-RNase genotypes inferred from PCR exactly matches the S-phenotypes deduced from test-crosses. Comparison of the sequences obtained with those of S-RNases already published allowed us to relate S-alleles between studies. This will allow the prediction of cross-incompatibility among an even larger number of european pear cultivars.

Sexual self-incompatibility in European pear (Pyrus communis L.) is controlled by a single locus (Slocus) 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 Salleles 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.

The stylar products of the S-locus for the gametophytic self-incompatibility (GSI) system in the Rosaceae are ribonucleases (S-RNases). Recently, sequences for 13 pear S-RNase alleles have been published and named following a letter-symbol nomenclature (S a to S d and S h to S p ). To establish the correspondence between these sequences and the self-incompatibility alleles we have described previously (S 1 to S 5 ), we have amplified genomic DNA with consensus primers from the cultivars, ÔWilliamsÕ (S 1 S 2 ), ÔCosciaÕ (S 3 S 4 ), ÔButirra Precoce MorettiniÕ (S 1 S 3 ), ÔSanta Maria MorettiniÕ (S 2 S 3 ) and ÔDoyenne du ComiceÕ (S 4 S 5 ) and identified PCR products specifically associated with each S allele. Cloning and sequencing of the amplification products has revealed that they correspond to European pear sequences already deposited in the database. This allowed us to link S-RNase sequences with S allele phenotypes and to determine a correspondence between the symbol-letter nomenclature used to name S-RNase sequences and the number-based nomenclature used to name S alleles. Based on this result the prediction of new cross-incompatibilities among pear cultivars is discussed. Finally, we propose a unified number-based nomenclature to avoid future confusion denominating S alleles in pear.