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Bolting is an important agronomic character of the Chinese cabbage, but premature bolting can greatly reduce its commercial value, yield, and quality. Here, early-bolting mutant 1 ( ebm1 ) was obtained from a Chinese cabbage doubled haploid (DH) line “FT,” by using an isolated microspore culture and ethyl methanesulfonate (EMS) mutagenesis. The  ebm1  was found to bolt extremely earlier than the wild type “FT.” Genetic analysis indicated that the phenotype of the  ebm1  was controlled by a single recessive nuclear gene. Using a mapping population of 1,502 recessive homozygous F 2  individuals with the  ebm1  phenotype, the  ebm1  gene was mapped to between the markers SSRhl-53 and SSRhl-61 on chromosome A04 by using SSR markers, and its physical distance was 73.4 kb. Seven genes were predicted in the target region and then cloned and sequenced; the only difference in the sequences of the  ebm1  and “FT” genes was with  Bra032169 . Unlike that in “FT,” the  Bra032169  in  ebm1  had a novel 53 bp insertion that caused the termination of amino acid coding. The mutation was not consistent with EMS mutagenesis, and thus, may have been caused by spontaneous mutations during the microspore culture. Based on the gene annotation information,  Bra032169  was found to encode the histone methyltransferase  CURLY LEAF  ( CLF ) in  Arabidopsis thaliana .  CLF  regulates the expression of flowering-related genes. Further genotyping revealed that the early-bolting phenotype was fully co-segregated with the insertion mutation, suggesting that  Bra032169  was the most likely candidate gene for  ebm1 . No significant differences were noted in the  Bra032169  expression levels between the  ebm1  and “FT.” However, the expression levels of the flowering-related genes  FLC ,  FT ,  AG , and  SEP3  were significantly higher in the  ebm1  than in the “FT.” Thus, the mutation of  Bra032169  is responsible for the early-bolting trait in Chinese cabbage. These results provide foundation information to help understand the molecular mechanisms of bolting in the Chinese cabbage.

frontiers in plant science

An Insertion Mutation in Bra032169 Encoding a Histone Methyltransferase Is Responsible for Early Bolting in Chinese Cabbage (Brassica rapa L. ssp. pekinensis)