Neofunctionalisation of the Sli gene leads to self-compatibility and facilitates precision breeding in potato

Genetic gain in potato is hampered by the heterozygous tetraploid genome of cultivated potato. Converting potato into a diploid inbred-line based F1-hybrid crop provides a promising route towards increased genetic gain. The introduction of a dominant S-locus inhibitor (Sli) gene into diploid potato germplasm allows efficient generation of self-fertilized seeds and thus the development of potato inbred lines. Little is known about the structure and function of the Sli locus. Here we describe the mapping of Sli to a 12.6 kb interval on chromosome 12 using a recombinant screen approach. One of two candidate genes present in this interval shows a unique sequence that is exclusively present in self-compatible lines. We describe an expression vector that converts self-incompatible genotypes into self-compatible and a CRISPR-Cas9 vector that converts SC genotypes into SI. The Sli gene encodes an F-box protein that is specifically expressed in pollen from self-compatible plants. A 533 bp insertion in the promotor of that gene leads to a gain of function mutation, which overcomes self-pollen rejection.

Production of inbred progenies of diploid potatoes using an S-locus inhibitor (Sli) gene, and their characterization

To develop inbred lines from self-incompatible, cultivated diploid potatoes, an S-locus inhibitor (Sli) gene derived from a self-compatible variant of a wild potato species, Solanum chacoense, was incorporated into various cultivated diploid potatoes. The progeny was selfed twice by the action of the Sli gene to obtain 74 S2 inbred clones belonging to 8 families. More than 40% of them were either non-flowering or pollen sterile. Among the pollen fertile clones, self-compatible clones occurred with a much lower frequency (20.9%) than expected (83.3%). The result demonstrated that self-compatibility was introduced and expressed in the gene pool of cultivated diploid potatoes by an action of the Sli gene, although serious inbreeding depression associated with selfing occurred. The genotypes of S2 inbreds were surveyed using 46 S. chacoense - specific RFLP (restriction fragment length polymorphism) markers covering the whole potato genome. More than half of the markers (67.4%) showed distorted segregation. Particularly, all markers on chromosome 12 were overrepresented in the S2 inbreds. This confirms our earlier finding that the Sli gene locates on chromosome 12 and the alleles linked with this gene are preferentially transmitted because of its essential requirement for selfing.Key words: RFLP markers, inbreds, diploid potato, distorted segregation.