Details

Autor(en) / Beteiligte

• Enciso-Rodriguez, Felix
• Manrique-Carpintero, Norma C
• Nadakuduti, Satya Swathi
• Buell, C Robin
• Zarka, Daniel
• Douches, David

Titel

Overcoming Self-Incompatibility in Diploid Potato Using CRISPR-Cas9

Ist Teil von

• Frontiers in plant science, 2019-04, Vol.10, p.376-376

Ort / Verlag

Switzerland: Frontiers Media S.A

Erscheinungsjahr

2019

Quelle

EZB Electronic Journals Library

Beschreibungen/Notizen

• Potato breeding can be redirected to a diploid inbred/F1 hybrid variety breeding strategy if self-compatibility can be introduced into diploid germplasm. However, the majority of diploid potato clones ( spp.) possess gametophytic self-incompatibility that is primarily controlled by a single multiallelic locus called the -locus which is composed of tightly linked genes, ( -locus RNase) and multiple ( -locus F-box proteins), which are expressed in the style and pollen, respectively. Using genes known to function in the Solanaceae gametophytic SI mechanism, we identified alleles with flower-specific expression in two diploid self-incompatible potato lines using genome resequencing data. Consistent with the location of the -locus in potato, we genetically mapped the gene using a segregating population to a region of low recombination within the pericentromere of chromosome 1. To generate self-compatible diploid potato lines, a dual single-guide RNA (sgRNA) strategy was used to target conserved exonic regions of the gene and generate targeted knockouts (KOs) using a Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9 (Cas9) approach. Self-compatibility was achieved in nine KO T lines which contained bi-allelic and homozygous deletions/insertions in both genotypes, transmitting self compatibility to T progeny. This study demonstrates an efficient approach to achieve stable, consistent self-compatibility through KO for use in diploid potato breeding approaches.