Flowering time is important for Brassica rapa vegetables because premature bolting before harvest can lower yield and quality. FLOWERING LOCUS C (FLC) acts as a key repressor of flowering. In this study, we identified a nonsynonymous mutation at the 58th nucleotide of exon1 in BrFLC1 (named as Pe1+58 (A/C)) by screening resequencing data of 199 B. rapa accessions and verified this mutation as being related to flowering time variation. Strong linkage inheritance was detected between this locus and a previously reported splicing site mutation at intron 6 of BrFLC1 (Pi6+1 (G/A)), showing as co-occurrence of BrFLC1Pe1+58(A) and BrFLC1Pi6+1(G), named as haplotype H1: AG, or co-occurrence of BrFLC1Pe1+58(C) and BrFLC1Pi6+1(A), named as haplotype H2: CA. The frequency distribution of BrFLC1 haplotypes skewed to the haplotype H1 in turnip, broccoletto, mizuna, komatsuna, and taicai, while it was skewed to the haplotype H2 in caixin, pak choi, zicaitai, and wutacai. The frequencies of the two haplotypes were comparable in Chinese cabbage. This indicated that BrFLC1 haplotypes were related to B. rapa intraspecific diversification. Further analysis of a Chinese cabbage collection revealed that accessions from the spring ecotype preferred to keep H1: AG and almost all accessions from the summer ecotype were H2: CA. The early flowering haplotype of BrFLC1 was purified in summer Chinese cabbage, indicating that BrFLC1 had been strongly selected during genetic improvement of summer Chinese cabbages. A significant difference in flowering time of F2 individuals with the homologous BrFLC1Pi6+1(G) allele but different BrFLC1Pe1+58 (A/C) alleles, indicated that this locus had independent genetic effects on flowering time. The newly identified allelic diversity of BrFLC1 can be used for breeding of resistance to premature bolting in B. rapa vegetables.
Multiplex CRISPR/Cas9 Mutagenesis of BrVRN1 Delays Flowering Time in Chinese Cabbage (Brassica rapa L. ssp. pekinensis)
