Lobed leaves are advantageous for gas exchange, canopy architecture, and high-density planting; however, the genetic mechanisms of leaf lobe formation in Brassica crops remains poorly understood. Here, lob10.1, our previously identified major QTL controlling the presence/absence of leaf lobes in B. rapa (AA), was fine mapped to a confidence interval of 69.8 kb. REDUCED COMPLEXITY ORGAN (BrRCO, BraA10g032440.3c), a homeodomain leucine zipper class I (HD ZIP I) transcription factor, was predicted to be the most likely candidate gene underlying lob10.1. Null mutations of BrRCO by CRISPR/Cas9 in the lobed-leaf parent RcBr and over-expression in the counter-part near isogenic lines (NILRcBr) lead to entire and lobed leaves, respectively. Analysis of the gene evolution revealed that A10. RCO functions as a core gene and was generally negatively selected in B. rapa. Moreover, BrRCO function as a negative regulator by directly binding to promoters of BrACP5 and repressing its expression. The function of ACID PHOSPHATASE TYPE 5 (BrACP5) was subsequently confirmed as VIGS-BrACP5 produced entire leaves in RcBr. This study identified the core gene BrRCO to be involved in the development of leaf lobes in B. rapa and elucidated a new pathway for leaf lobe formation by the BrRCO-BrACP5 module. These findings provide a theoretical basis for the formation of leaf lobes in Brassica crops.

Leaf shape is an important determinant of photosynthesis, yield and quality in plants. In this study, we obtained a curled leaf mutant, cl1, from an ethyl methanesulfonate (EMS)-induced mutagenesis population. It was designated the Brcl1^YS locus. Bulk segregant RNA sequencing combined with recombinant screening identified the candidate interval responsible for Brcl1^YS in a 97.5 kb region on chromosome A02. Twelve genes were identified within the candidate region. Sequence differences and co-separation verification confirmed that BraA02g017030.3C was the most promising candidate gene underlying the Brcl1^YS locus. It is homologous to Arabidopsis AT1G66350 (RGL1), which has been shown to act as a negative regulator of the gibberellin pathway. Combined with cell morphology observation, it is speculated that the loss of function of Brcl1^YS results in differences in cell development, ultimately leading to changes in leaf morphology. The results will contribute to the understanding of the molecular mechanisms underlying leaf curling in B. rapa.

In crops such as Brassica rapa, the agronomic trait plant height affects the leaf light absorption rate, benefits substance accumulation, and increases production by increasing the biological yield of the population. The mechanism of plant height was adequately studied in Brassica napus and Arabidopsis, while Brassica rapa had been rarely reported. Herein, we studied a B. rapa dwarf mutant Brcd1^YS. Compared with its wild-type, Yellow sarson, Brcd1^YS showed shorter hypocotyls and slow growth, with inhibited root elongation and reduced numbers of lateral roots. Chlorophyll content determination and pollen viability detection indicated that the mutant’s chlorophyll content was higher than that of the wild-type; however, its pollen was inactive. Cytological identification showed that the number of cells in Brcd1^YS leaves was significantly reduced and were arranged irregularly compared with those of the wild-type. Bulked Segregant RNA sequencing combined with conventional linkage mapped the dwarf mutation Brcd1^YS to Chromosome A06, at position 21,621,766 to 24,683,923 bp. Application of exogenous gibberellic acid (GA) partially rescued the dwarf phenotype of Brcd1^YS. GA-related genes BraA06g034110.3C and BraA06g033010.3C were identified as the most promising candidate genes. These results make a valuable contribution to our understanding of the mechanism of plant height determination in B. rapa, paving the way for further breeding of dwarf B. rapa.

In Brassica rapa, flowering time (FT) is an important agronomic trait that affects the yield, quality, and adaption. FT a complicated trait that is regulated by many genes and is affected greatly by the environment. In this study, a chromosome segment substitution line (CSSL), CSSL16, was selected that showed later flowering than the recurrent parent, rapid-cycling inbred line of B. rapa (RcBr). Using Bulked Segregant RNA sequencing, we identified a late flowering quantitative trait locus (QTL), designated as qFT7.1, on chromosome A07 based on a secondary-F₂ population derived from the cross between CSSL16 and RcBr. qFT7.1 was further validated by conventional QTL mapping. This QTL explained 39.9% (logarithm of odds = 32.2) of the phenotypic variations and was fine mapped to a 56.4-kb interval using recombinant analysis. Expression analysis suggests that BraA07g018240.3C, which is homologous with ATC (encoding Arabidopsis thaliana CENTRORADIALIS homologue), a gene for delayed flowering in Arabidopsis as the most promising candidate gene. Sequence analysis demonstrated that two synonymous mutations existed in the coding region and numerous bases replacements existed in promoter region between BraA07g018240.3C from CSSL16 and RcBr. The results will increase our knowledge related to the molecular mechanism of late flowering in B. rapa, and lay a solid foundation for the breeding of late bolting in B. rapa.