Overcoming Self-Incompatibility in Diploid Potato Using CRISPR-Cas9

Area of Science:

• Plant genetics
• Molecular biology
• Agricultural science

Background:

• Diploid potato breeding is limited by gametophytic self-incompatibility.
• Self-incompatibility is controlled by the S-locus, comprising S-RNase and SLF genes.

Purpose of the Study:

• To develop self-compatible diploid potato lines for advanced breeding.
• To investigate the role of S-RNase in potato self-incompatibility.

Main Methods:

• Identified S-RNase alleles using genome resequencing.
• Mapped the S-RNase gene to chromosome 1 using genetic segregation.
• Utilized CRISPR-Cas9 with dual sgRNAs to knockout S-RNase in diploid potatoes.

Main Results:

• Successfully generated nine S-RNase knockout (KO) T0 lines exhibiting self-compatibility.
• KO lines showed bi-allelic and homozygous deletions/insertions.
• Self-compatibility was stably transmitted to T1 progeny.

Conclusions:

• Achieved stable and consistent self-compatibility in diploid potatoes via S-RNase KO.
• This method provides an efficient tool for diploid potato breeding programs.
• Facilitates the development of inbred/F1 hybrid varieties.