Identification of S-RNase Genotypes of 65 Almond [Prunus dulcis (Mill.) D.A. Webb] Germplasm Resources and Close Relatives

Self-incompatibility (SI) systems in plants prevent self-pollination and mating among relatives, enhancing genetic diversity in nature but posing challenges in almond production and breeding. S-allele composition alongside the flowering periods of these cultivars enables the anticipation of cross-compatibility and optimal cultivar combinations for the allocation of pollinating trees in production. In the current study, 65 materials containing 61 almond (Prunus dulcis) germplasm resources, of which two were hybrids and the remaining four were peach (Prunus persica) germplasms, were used for the S-RNase genotype. The results showed that 55 genomic samples were amplified by PCR to obtain double-banded types, which identified their complete S-RNase genotypes, while the rest of the samples amplified only a single band, identifying one S-RNase gene in the S gene. A total of 30 S-RNase genes were identified in Prunus dulcis, Prunus webbii, Prunus persica, Prunus armeniaca, Prunus salicina, and Prunus cerasifera. Sequence analysis revealed polymorphisms spanning from 313 to 2031 bp within the amplified fragment sequence. The S57-RNase gene exhibited the highest frequency at 31.75% among the identified materials, with S1S57, S10S57, and S7S57 being the predominant S genotypes. A new S-RNase gene, named S65, was identified with a sequencing length of 1483 bp. Its deduced amino acid sequence shared 98.24% similarity with the amino acid sequence of the S-RNase gene on GenBank, with the highest homology. Furthermore, according to the findings, 65 materials belong to eight S genotype cross-incompatibility groups (CIG) and one semi-compatibility or compatibility group (0). Among them, most of the seven main almond germplasm resources and 35 cultivars can be cross-pollinated. The results of the study can lay the foundation for pollinator tree allocation and breeding hybrid parent selection in almond production.