Rapeseed (Brassica napus, B. napus) is an important oil seed crop in the world. Previous studies showed that seed germination vigor might be correlated with seed oil content in B. napus, but the regulation mechanism for seed germination has not yet been explained clearly. Dissecting the regulation mechanism of seed germination and germination vigor is necessary. Here, proteomic and genomic approaches were used to analyze the germination process in B. napus seeds with different oil content. The identification of 165 differentially expressed proteins (DEPs) in the germinating seeds of B. napus with high and low oil content was accomplished by two-dimensional fluorescence difference in gel electrophoresis (2D-DIGE). The comparative proteomic results revealed that seeds with high oil content had higher metabolic activity, especially for sulfur amino acid metabolism. Thirty-one unique genes were shown to be significantly changed during germination between the seeds with high and low oil content, and thirteen of these genes were located within the confidence interval of germination-related quantitative trait locus (QTLs), which might play an important role in regulating seed germination vigor. The present results are of importance for the understanding of the regulation mechanism for seed germination vigor in B. napus.

Rapeseed (Brassica napus) is an important oil seed crop in the Brassicaceae family. Chemical induced male sterility (CIMS) is one of the widely used method to produce the hybrids in B. napus. Identification of the key genes and pathways that involved in CIMS were important to understand the underlying molecular mechanism. In the present report, a multi-omics integrative analysis, including of the proteomic, transcriptomic and miRNAs, combined with morphological and physiological analysis were conducted. Earlier degeneration of the tapetosomes and elaioplasts, aberrantly stacking in tapetal cells and incompletely deposition in tryphine of pollen wall were observed in chemical hybridization agent (CHA) of SX-1 treated B. napus through SEM and TEM analysis. It was revealed that the deficiencies in protein processing in endoplasmic reticulum (ER) and flavonoids biosynthesis were occurred at early stage in the SX-1 treated materials. Subsequently, plant hormone signal transduction, biosynthesis of amino acids, fatty acids and steroid in anther at later stages were identified down-regulated after SX-1 treatment. 144 transcript factors (TFs) were also indentified to down-regulated at early stage, which suggested the early regulation in anther and pollen wall development were disordered in CHA treated B. napus. In addition, 7 important miRNAs were identified and 2 of the predicted target genes of miRNAs were Rf-like genes. Taken together, an interaction network of candidate genes and the putative metabolism pathways were constructed based on the multi-omics integrative analysis, it provided a new insight into the male sterility induced by CHA of SX-1 in B. napus.

Varieties with a semi-dwarf compact plant architecture may increase yield per unit area in rapeseed (Brassica napus) by allowing high-density cultivation and mechanical harvesting while conferring lodging resistance. Mutation of ERECTA (ER), which encodes a receptor-like protein kinase, generates a compact and upright plant architecture in Arabidopsis thaliana; however, there have been no reports on the roles of the ER family (ERf) in B. napus. In this study, we used the CRISPR/Cas9 system to generate mutants in each of the two homoeologs of B. napus ERf members BnaER and ER-Like 1 (BnaERL1), and in the single BnaERL2 gene, resulting in the homozygous mutants BnaA09.er/BnaC08.er, BnaA06.erl1/BnaC03.erl1, and BnaA10.erl2. Under greenhouse conditions, BnaA09.er/BnaC08.er plants were shorter than the wild type, with a compact inflorescence and shorter siliques. In addition, BnaA09.er/BnaC08.er plants produced significantly more branches and total siliques than the wild type, with no significant changes in the number of ovules per silique or thousand-seed weight. Under field conditions, the BnaA09.er/BnaC08.er mutant plant showed a phenotype comparable to that under greenhouse conditions, but with a notable drop in thousand-seed weight. These results indicate that the BnaA09.er/BnaC08.er mutant offers a valuable germplasm resource for breeding rapeseed with ideal plant architecture. The online version contains supplementary material available at 10.1007/s42994-025-00217-4.

Enhancing rapeseed (Brassica napus, B. napus) yield is critical for ensuring global vegetable oil security. However, yield is heavily influenced by silique development and seed size, the enhancement of which is limited by scarce genetic resources. The CRISPR/Cas9 system has emerged as a powerful tool for constructing genome-wide mutant libraries, even in polyploid crops with complex genomes. The transcriptome-wide association study (TWAS) data, tissue-specific expression profiles data and reported genes were integrated to identify candidate genes regulating silique development and seed size. We constructed a sgRNA library targeting these genes and generated a CRISPR/Cas9 editing mutant library through genetic transformation. Specifically, 6124 sgRNAs were designed for 1739 candidate genes with ≦ 4 orthologues. 681 T0 plants were obtained through genetic transformation, which harbor 453 sgRNAs. Of 408 T0 plants analyzed, 151 (37.00%) exhibited successful gene editing events, targeting 84 candidate genes. Ten homozygous mutant plants were isolated and preliminary phenotypic analysis was performed in mutants targeting the BnaHRDs. The results suggest that mutations in BnaHRD.A03 and BnaHRD.C03 may modulate plant height (PH), main inflorescence length (MIL), silique length (SL), effective silique number per plant (ENS), seed number per silique (SNPS), and thousand-seed weight (TSW). This study harnessed the CRISPR/Cas9 technology to establish a preliminary library of gene-edited mutants in B. napus, thereby laying a robust foundation for the future screening of candidate genes pertaining to silique development and seed size. Furthermore, this study provides a methodological framework for rapid functional gene discovery in B. napus through CRISPR-based approaches.

The microtubule-associated protein 65 (MAP65) family comprises key plant cytoskeletal regulators essential for microtubule array organization, cell division, cell elongation, and organ morphogenesis. Recent advances in molecular and cellular biology have facilitated functional characterization of these proteins. This review systematically summarizes the structural features, subcellular localization, action mechanisms, regulatory networks, and functional divergence across species. Emerging evidence indicates that MAP65s not only participate in conventional microtubule functions, but also precisely regulate cell division orientation and fruit morphogenesis through coordinated interactions with other protein families. Moreover, MAP65s serve as important targets of pathogen effectors. This review further discusses the frontier areas in MAP65 research, including structural biology, dynamic regulatory mechanisms, and potential applications in crop improvement.