Mutating via CRISPR-Cas9 increases the seed size and weight in .

Unlabelled: Seed weight, which is highly correlated to seed size, is a critical agronomic trait that determines the yield of However, there have been limited researches on the genes involved in regulating seed size. In , (), an E3 ubiquitin ligase gene, has been identified as a significant negative regulator in controlling organ size, but the function of its homologs in rapeseed remains unknown. Only two homologous of , , have been found in and were mutated using the CRISPR-Cas9 system. Three T-DNA-free lines, T-157-1-C8, T-390-2-B8, and T-397-2-E2, were identified from the homozygous T mutant lines. The BnaEOD1.A04 showed a similar type of editing in these mutants, whereas the BnaEOD1.C04 in T-397-2-E2 was only missing 26 amino acids, and the translation was not prematurely terminated, which was different from the other two mutants. In parallel, mutation of resulted in a noteworthy increase in both seed size and seed weight in the three editing lines. Additionally, there was a significant decline in the number of seeds per silique (SPS) and silique length (SL) in T-157-1-C8 and T-390-2-B8, but T-397-2-E2 did not show any significant changes in the SPS and SL, possibly due to distinct types of editing in the three lines. The above results indicate the conserved function of homologs and provides promising germplasm for breeding novel high-yield rapeseed varieties by improving seed size and thousand-seed weight.

Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-023-01430-z.