Long-read genome sequencing reveals the sequence characteristics of pear self-incompatibility locus

The S-RNase-based self-incompatibility locus (S-locus) in Petunia species contains 16-20 F-box genes, which collaboratively function in the recognition and subsequent degradation of non-self S-RNases, while distinguishing them from self S-RNase. However, the number of S-locus F-box genes (SFBBs) physically interacted with non-self S-RNases remains uncertain in Pyrus species. Utilizing Pacbio long-read sequencing, we successfully assembled the genome of pear cultivar 'Yali' (Pyrus bretschneideri), and identified 19 SFBBs from the Pyrus S₁₇-locus spanning approximately 1.78 Mb. Additionally, we identified 17-21 SFBBs from other Pyrus and Malus S-loci spanning a range of 1.35 to 2.64 Mb. Based on the phylogenetic analysis, it was determined that Pyrus and Malus SFBBs could be classified into 22 groups, denoted as I to XXII. At amino acid level, SFBBs within a given group exhibited average identities ranged from 88.9% to 97.9%. Notably, all 19 SFBBs from the S₁₇-locus co-segregated with S₁₇-RNase, with 18 of them being specifically expressed in pollen. Consequently, these 18 pollen-specifically expressed SFBBs are considered potential candidates for the pollen-S determinant. Intriguingly, out of the 18 pollen-specifically expressed SFBBs, eight demonstrated interactions with at least one non-self S-RNase, while the remaining SFBBs failed to recognize any S-RNase. These findings provide compelling evidence supporting the existence of a collaborative non-self-recognition system governing self-incompatibility in pear species.