Clubroot, a devastating soil borne disease affecting 30%∼50% of Brassicaceae crops worldwide, lacks effective control measures. In the present study, we explored the potential of melatonin (MT) and copper oxide nanoparticle (CuO-NPs) in mitigating clubroot severity in the Brassica rapa ssp. pekinensis. Following 18 h priming with MT, CuO-NPs, or both seeds were grown in controlled environment using synthetic potting mix. Inoculated with Plasmodiophora brassicae spores on 5th day, followed by a soil drench phyto-nano treatment with a week interval. Plants were assessed for various health and growth indices including disease, biometrics, photosynthesis, reactive oxygen species (ROS), antioxidant enzyme activity, hormones and genes expression at onset of secondary clubroot infection using established protocols. Statistical analysis employed ANOVA with Fisher's LSD for significance assessment (P < 0.05). Our results revealed that seed priming with both MT (50 μMol/L) and CuO-NPs (200 mg/L), followed by soil drenching significantly reduced clubroot incidence (38%) and disease index (57%), compared to control treatments. This synergistic effect was associated with enhanced plant growth (shoots: 48% and roots: 59%). Plants treated with both MT and CuO-NPs showed robust antioxidant defenses, significantly increased superoxide dismutase (SOD (25/29%)), catalase (CAT (83/55%)), and ascorbate peroxidase (APX (83/46%)) activity in both shoots/roots, respectively, compared to infected control. Notably, salicylic acid and jasmonic acid levels doubled in treated plants, while stress hormone abscisic acid (ABA) decreased by 80% in roots and 21% in shoots. Gene expression analysis corroborated these findings, showing that the combined treatment activated antioxidant defense genes (SOD, APX and CAT) by 1.9-7.2-fold and upregulated hormone signaling genes JAZ1 (7.8-fold), MYC2 (3.9-fold) and SABP2 (36-fold). Conversely, ABA biosynthesis genes (ABA1 and NCED1) were downregulated up to 7.2-fold, while plant resistance genes NPR1, PRB1 and PDF1.2 were dramatically increased by up to 6.3-fold compared to infected plants. Overall, our combined treatment approach significantly reduces clubroot severity in B. rapa via enhanced antioxidant defenses, improved ROS scavenging, coordinated hormonal regulation and increased pathogen response genes. This study offers promising strategy for developing effective control measures against clubroot in susceptible cruciferous crops.

Clubroot, caused by Plasmodiophora brassicae, is a destructive soil-borne disease significantly harming global Brassica crop production. This study employed the Williams and European Clubroot Differential (ECD) and Williams systems to identify the pathotypes of P. brassicae collected from Hangzhou City, Yuhang District and Quzhou City, Kaihua County in Zhejiang Province. Greenhouse and field trials were conducted to evaluate the effects of plastic film covering and four chemical agents on the growth parameters and clubroot severity of the Chinese cabbage cultivar ‘Granaat’. Potential treatment mechanisms on clubroot were explored through a qPCR analysis of the resting spore density and pH measurement of the soil. Furthermore, treatment with 1-napthaleneacetic acid (NAA), a synthetic auxin, was also evaluated for its potential role in suppressing clubroot. The results indicate that the pathotypes of P. brassicae in the two districts were P1, ECD20/31/12, and P3, ECD20/15/4. While an individual application of plastic film covering could not effectively control clubroot, calcium cyanamid, dazomet and ammonium bicarbonate demonstrated significant efficacy in its management. These three agents significantly reduced the resting spore density in the soil, with calcium cyanamid and ammonium bicarbonate also increasing soil alkalinity. Additionally, ammonium bicarbonate promotes lateral root development in ‘Granaat,’ helping infected plants access adequate water and nutrients. However, NAA exhibited no efficacy in clubroot control. Therefore, sustained lateral root development is crucial for effectively resisting P. brassicae invasion. Considering application costs and environmental friendliness, we propose the field application of ammonium bicarbonate as the optimal method for clubroot disease management in Zhejiang Province.

Flowers are one of the most important organs in plants. Their development serves as a key indicator of the transition from vegetative to reproductive growth and is regulated by various internal signals and environmental factors. NAC (NAM, ATAF, CUC) transcription factors (TFs) play a crucial regulatory role in floral organ development; however, research on the analysis and identification of the NAC TF family in Chinese cabbage (Brassica rapa L.) remains limited. In this study, we performed a comprehensive genome-wide analysis of NACs in B. rapa and identified 279 members of the BrNAC gene family. Their physicochemical properties, domain structure, collinearity relation, and cis-regulatory elements were evaluated. Phylogenetic analysis indicates that NAC proteins from Arabidopsis, B. rapa, B. oleracea, and B. nigra can be classified into seven distinct clades. BrNACs exhibit a tissue-specific expression, and nine BrNACs being specifically expressed in the inflorescence. Furthermore, nine flower-related BrNACs were selected for RT-qPCR analysis to validate their expression profiles. BrNAC2s has been cloned to investigate their subcellular localization, and examine the expression patterns of their promoters in Arabidopsis inflorescences. BrNAC2a and BrNAC2c are highly expressed in stamens while BrNAC2b exhibits elevated expression in pistils and pedicel. Collectively, our findings enhance the understanding of the BrNAC family and provide a foundation for future studies on the molecular mechanisms of BrNACs in floral development.

Simple sequence repeats (SSRs), also known as microsatellites, stand out as the most crucial molecular markers in both animals and plants owing to their high polymorphism, extensive information content, ease of detection through polymerase chain reaction (PCR) assays, and widespread distribution across the genome. In this study, a total of 125,443 SSR loci were identified from the whole-genome sequence of B. oleracea, and 82,948 primer pairs for SSR have been designed. Furthermore, each primer pair is designated with a unique identifier (ranging from BolSSR00001 to BolSSR82984). Our findings indicated that certain markers within them could be transferred to other cruciferous crops. In addition, a total of 336 pairs of SSR primers have been used to screen the polymorphism between the bolting-resistant and bolting-easy gene pools. After the test of verification with F₂ generation individual plants, we obtained an SSR dominant marker, BolSSR040196, linked with bolting-resistant locus in cabbage, and the genetic distance between this SSR marker and the bolting-resistant locus was 10.69 cM. Moreover, BolSSR040196 is located on the C05 chromosome with a CT motif, characterized by a repeat of 9 in bolting-easy plants and 11 in bolting-resistant plants. Haplotype analysis showed that the correct prediction rate reached 82.35%. The BolSSR040196 marker can be used in marker-assisted selection (MAS) breeding, offering a straightforward and efficient approach for bolting-resistant cabbage breeding in the future.

MADS-box is a vital transcription factor family that functions in plant growth and development. Apart from APETALA2, all genes in the ABCDE model that explain the molecular mechanism of floral organ development belong to the MADS-box family. Carpel and ovule numbers in plants are essential agronomic traits that determine seed yield, and multilocular siliques have great potential for the development of high-yield varieties of Brassica. In this study, ABCDE genes in the MADS-box family from Brassica rapa were identified and characterized. Their tissue-specific expression patterns in floral organs and their differential expression in different pistil types of B. rapa were revealed by qRT-PCR. A total of 26 ABCDE genes were found to belong to the MADS-box family. Our proposed ABCDE model of B. rapa is consistent with that of Arabidopsis thaliana, indicating that ABCDE genes are functionally conserved. These results of qRT-PCR showed that the expression levels of class C and D genes were significantly different between the wild-type (wt) and tetracarpel (tetrac) mutant of B. rapa. Interestingly, the expression of the homologs of class E genes was imbalanced. Therefore, it is speculated that class C, D, and E genes are involved in developing the carpel and ovule of B. rapa. Our findings reveal the potential for the selection of candidate genes to improve yield traits in Brassica crops.

Brassica napus L. is a major oilseed crop whose productivity is significantly affected by abiotic stresses such as drought. PYR/PYL/RCAR (PYL) proteins act as key abscisic acid (ABA) receptors and play central roles in stress responses. However, a comprehensive genome-wide analysis of the PYL gene family in B. napus is still lacking, limiting our understanding of their functions in plant and stress adaptation. This study reports the first comprehensive genome-wide analysis of the PYL gene family in B. napus (rapeseed), cultivar ZS11, identifying 25 BnPYL genes grouped into four subfamilies, I (four genes), I-II (five genes), II (five genes), III (11 genes), and their encoded proteins were predicted to be mainly localized in the chloroplast. Structural analysis revealed diverse exon–intron organization and 10 conserved motifs. All identified BnPYLs contained Polyketide_cyc2 domains (PF10604), supporting their annotation as members of the PYL family. Promoter analysis identified cis-regulatory elements related to light response, stress regulation, and hormonal signaling. Computational analysis of post-translational modifications suggested that phosphorylation sites are mainly localized at serine and threonine residues. Tertiary structure modelling revealed conserved three-dimensional architectures among BnPYL proteins, suggesting potential functional conservation. Expression profiling and RT-qPCR analyses revealed that several BnPYL genes respond to ABA-mediated drought stress, with BnPYL15 and BnPYL22 exhibiting the highest induction (4–5-fold) and BnPYL2, BnPYL5, BnPYL6, BnPYL17, BnPYL18, and BnPYL25 showing significant upregulation (2.0–4.5-fold), suggesting potential roles in enhancing drought tolerance in B. napus.