The plant circadian clock temporally drives gene expression throughout the day and coordinates various physiological processes with diurnal environmental changes. It is essential for conferring plant fitness and competitive advantages to survive and thrive under natural conditions through the circadian control of gene transcription. Chinese cabbage (Brassica rapa ssp. pekinensis) is an economically important vegetable crop worldwide, although there is little information concerning its circadian clock system. Here we found that gene expression patterns are affected by circadian oscillators at both the transcriptional and post-transcriptional levels in Chinese cabbage. Time-course RNA-seq analyses were conducted on two short-period lines (SPcc-1 and SPcc-2) and two long-period lines (LPcc-1 and LPcc-2) under constant light. The results showed that 32.7–50.5% of the genes were regulated by the circadian oscillator and the expression peaks of cycling genes appeared earlier in short-period lines than long-period lines. In addition, approximately 250 splicing events exhibited circadian regulation, with intron retention (IR) accounting for a large proportion. Rhythmically spliced genes included the clock genes LATE ELONGATED HYPOCOTYL (BrLHY), REVEILLE 2 (BrRVE2) and EARLY FLOWERING 3 (BrELF3). We also found that the circadian oscillator could notably influence the diurnal expression patterns of genes that are associated with glucose metabolism via photosynthesis, the Calvin cycle and the tricarboxylic acid (TCA) cycle at both the transcriptional and post-transcriptional levels. The collective results of this study demonstrate that circadian-regulated physiological processes contribute to Chinese cabbage growth and development.

Protoplast-based transient gene expression system has been widely used in plant genome editing because of its simple operation and less time-consuming. In order to establish a universal protoplast-based transient transfection system for verifying activities of genome editing vectors containing targets in Brassica, we systematically optimized factors affecting protoplast isolation and transient gene expression. We established an efficient protoplast-based transient gene expression system (PTGE) in Chinese cabbage, achieving high protoplast yield of 4.9 × 10⁵ · g⁻¹ FW, viability over 95%, and transfection efficiency of 76%. We showed for the first time that pretreatment of protoplasts with a hypotonic MMG could significantly enhance the transfection efficiency. Furthermore, protoplasts incubated at 37 ℃ for 6 min improved the transfection efficiency to 86%. We also demonstrated that PTGE worked well (more than 50% transfection efficiency) in multiple Brassica species including cabbage, Pak Choi, Chinese kale, and turnip. Finally, PTGE was used for validating the activities of CRISPR/Cas9 vectors containing targets in Chinese cabbage, cabbage, and pak choi, demonstrating the broad applicability of the established PTGE for genome editing in Brassica crops.

Properly regulated flowering time is pivotal for successful plant reproduction. The floral transition from vegetative growth to reproductive growth is regulated by a complex gene regulatory network that integrates environmental signals and internal conditions to ensure that flowering takes place under favorable conditions. Brassica rapa is a diploid Cruciferae species that includes several varieties that are cultivated as vegetable or oil crops. Flowering time is one of the most important agricultural traits of B. rapa crops because of its influence on yield and quality. The transition to flowering in B. rapa is regulated by several environmental and developmental cues, which are perceived by several signaling pathways, including the vernalization pathway, the autonomous pathway, the circadian clock, the thermosensory pathway, and gibberellin (GA) signaling. These signals are integrated to control the expression of floral integrators BrFTs and BrSOC1s to regulate flowering. In this review, we summarized current research advances on the molecular mechanisms that govern flowering time regulation in B. rapa and compare this to what is known in Arabidopsis.

Chinese cabbage (Brassica rapa ssp. pekinensis) has a long cultivation history and is one of the vegetable crops with the largest cultivation area in China. However, salt stress severely damages photosynthesis and hormone metabolism, nutritional balances, and results in ion toxicity in plants. To better understand the mechanisms of salt-induced growth inhibition in Chinese cabbage, RNA-seq and physiological index determination were conducted to explore the impacts of salt stress on carbon cycle metabolism and photosynthesis in Chinese cabbage. Here, we found that the number of thylakoids and grana lamellae and the content of starch granules and chlorophyll in the leaves of Chinese cabbage under salt stress showed a time-dependent response, first increasing and then decreasing. Chinese cabbage increased the transcript levels of genes related to the photosynthetic apparatus and carbon metabolism under salt stress, probably in an attempt to alleviate damage to the photosynthetic system and enhance CO₂ fixation and energy metabolism. The transcription of genes related to starch and sucrose synthesis and degradation were also enhanced; this might have been an attempt to maintain intracellular osmotic pressure by increasing soluble sugar concentrations. Soluble sugars could also be used as potential reactive oxygen species (ROS) scavengers, in concert with peroxidase (POD) enzymes, to eliminate ROS that accumulate during metabolic processes. Our study characterizes the synergistic response network of carbon metabolism and photosynthesis under salt stress.

Flowering time is an important agronomic trait of Chinese cabbage with late flowering being a primary breeding objective. In our previous work, we obtained Chinese cabbage-cabbage translocation lines that contained several beneficial cabbage genes. Cabbage-specific molecular markers show that these genes were coming from chromosome C01 of cabbage. In this study, we investigated the inheritance of flowering time in a couple of translocation lines and analyzed the transmission rate of molecular markers in the offspring. Consequently, we obtained the late flowering Chinese cabbage-cabbage translocation line 'AT7–4' in which the flowering time was later than that of '85–1' by about 7 days under 4-week vernalization. Based on previous studies of the genomes of Chinese cabbage and cabbage, we located the cabbage-specific molecular markers that were closely linked at the top of the chromosome A01 in the F₂ mapping population generated by self-crossing F₁s derived from a cross between the translocation line 'AT7–4' and Chinese cabbage '14–36'. Five flowering-related genes in the alien fragment were found by functional annotation and their molecular markers were developed. This study lays the foundation for the future improvement of Chinese cabbage varieties using A-C translocation lines.