Non-heading Chinese cabbage (NHCC) is an important leafy vegetable cultivated worldwide. Here, we report the first high-quality, chromosome-level genome of NHCC001 based on PacBio, Hi-C, and Illumina sequencing data. The assembled NHCC001 genome is 405.33 Mb in size with a contig N50 of 2.83 Mb and a scaffold N50 of 38.13 Mb. Approximately 53% of the assembled genome is composed of repetitive sequences, among which long terminal repeats (LTRs, 20.42% of the genome) are the most abundant. Using Hi-C data, 97.9% (396.83 Mb) of the sequences were assigned to 10 pseudochromosomes. Genome assessment showed that thisB. rapaNHCC001 genome assembly is of better quality than other currently availableB. rapaassemblies and that it contains 48,158 protein-coding genes, 99.56% of which are annotated in at least one functional database. Comparative genomic analysis confirmed thatB. rapaNHCC001 underwent a whole-genome triplication (WGT) event shared with otherBrassicaspecies that occurred after the WGD events shared withArabidopsis. Genes related to ascorbic acid metabolism showed little variation among the threeB. rapasubspecies. The numbers of genes involved in glucosinolate biosynthesis and catabolism were higher in NHCC001 than in Chiifu and Z1, due primarily to tandem duplication. The newly assembled genome will provide an important resource for research onB. rapa, especiallyB. rapassp.chinensis.

Salinity is a universal environmental stress that causes yield reduction in plants. WRKY33, which has been extensively studied in plant defense against necrotrophic pathogens, has recently been found to be important in salt-responsive pathways. However, the underlying molecular mechanisms controlling the involvement of WRKY33 in salt tolerance have not been fully characterized. Here, we explored the function of BcWRKY33A in non-heading Chinese cabbage (NHCC). Under salt stress, BcWRKY33A expression is significantly induced in roots. As a nuclear protein, BcWRKY33A has strong transcriptional activation activity. Overexpression of BcWRKY33A confers salt tolerance in Arabidopsis, whereas silencing of BcWRKY33A causes salt sensitivity in NHCC. Furthermore, BcHSFA4A, a protein that interacts with BcWRKY33A, could directly bind to the HSE motif within the promoters of BcZAT12 and BcHSP17.6A, which are involved in the plant response to salt stress. Finally, we found that BcWRKY33A could enhance the transcriptional activity of BcHSFA4A and affect its downstream genes (e.g. BcZAT12 and BcHSP17.6A), and co-overexpression of BcWRKY33A and BcHSFA4A could promote the expression of salt-related genes, suggesting that the regulatory interaction between BcWRKY33A and BcHSFA4A improves salt tolerance in plants. Overall, our results provide insight into the molecular framework of the BcWRKY33A-BcHSFA4A signaling pathway, which also aids in our understanding of the molecular mechanism of salt tolerance in plants.

A method using Agrobacterium rhizogenes-mediated callus production and plant regeneration enables efficient genetic transformation and gene editing in Chinese cabbage.

As a reducing substance, ascorbic acid functioned well in abiotic and biotic stress. However, the regulatory mechanism of drought resistance is rarely known in pak choi. Here we found a gene BcSRC2 containing a C2 domain that responds to ABA signal and drought regulation in pak choi. Silencing of BcSRC2 reduces ascorbic acid content and drought resistance of pak choi. In Arabidopsis, BcSRC2 overexpression promotes ascorbic acid accumulation and increases drought tolerance. Meanwhile, transcriptome analysis between WT and BcSRC2-overexpressing pak choi suggests that ascorbic acid-related genes are regulated. BcSRC2 interacts with BcAPX4 and inhibit APX activity in vitro and in vivo, increasing the ascorbic acid content. We also found that drought stress increases ABA content, which reduces the expression of BcMYB30. BcMYB30 bound to the promoter of BcSRC2 and reduced its expression. Overall, our results suggest that a regulatory module, BcMYB30-BcSRC2-BcAPX4, plays a central role in increasing ascorbic acid content for responding ABA-mediated drought regulation in pak choi.