The formation and evolution of flower coloration in Brassica crops.

The flower coloration of Brassica crops possesses significant application and economic value, making it a research hotspot in the field of genetics and breeding. In recent years, great progress has been made in the research on color variation and creation of Brassica crops. However, the underlying molecular mechanisms and evolutional processes of flower colors are poorly understood. In this paper, we present a comprehensive overview of the mechanism of flower color formation in plants, emphasizing the molecular basis and regulation mechanism of flavonoids and carotenoids. By summarizing the recent advances on the genetic mechanism of flower color formation and regulation in Brassica crops, it is clearly found that carotenoids and anthocyanins are major pigments for flower color diversity of Brassica crops. Meantime, we also explore the relationship between the emergence of white flowers and the genetic evolution of Brassica chromosomes, and analyze the innovation and multiple utilization of Brassica crops with colorful flowers. This review aims to provide theoretical support for genetic improvements in flower color, enhancing the economic value and aesthetic appeal of Brassica crops.

The story of a decade: Genomics, functional genomics, and molecular breeding in Brassica napus.

Rapeseed (Brassica napus L.) is one of the major global sources of edible vegetable oil and is also used as a feed and pioneer crop and for sightseeing and industrial purposes. Improvements in genome sequencing and molecular marker technology have fueled a boom in functional genomic studies of major agronomic characters such as yield, quality, flowering time, and stress resistance. Moreover, introgression and pyramiding of key functional genes have greatly accelerated the genetic improvement of important traits. Here we summarize recent progress in rapeseed genomics and genetics, and we discuss effective molecular breeding strategies by exploring these findings in rapeseed. These insights will extend our understanding of the molecular mechanisms and regulatory networks underlying agronomic traits and facilitate the breeding process, ultimately contributing to more sustainable agriculture throughout the world.

The mechanism of white flower formation in Brassica rapa is distinct from that in other Brassica species.

KEY MESSAGE: A single nucleotide (G) deletion in the third exon of BraA02.PES2-2 (Bra032957) leads to the conversion of flower color from yellow to white in B. rapa, and knockout mutants of its orthologous genes in B. napus showed white or pale yellow flowers. Brassica rapa (2n = 20, AA) is grown worldwide as an important crop for edible oil and vegetables. The bright yellow flower color and long-lasting flowering period give it aesthetic qualities appealing to countryside tourists. However, the mechanism controlling the accumulation of yellow pigments in B. rapa has not yet been completely revealed. In this study, we characterized the mechanism of white flower formation using a white-flowered natural B. rapa mutant W01. Compared to the petals of yellow-flowered P3246, the petals of W01 have significantly reduced content of yellowish carotenoids. Furthermore, the chromoplasts in white petals of W01 are abnormal with irregularly structured plastoglobules. Genetic analysis indicated that the white flower was controlled by a single recessive gene. By combining BSA-seq with fine mapping, we identified the target gene BraA02.PES2-2 (Bra032957) homologous to AtPES2, which has a single nucleotide (G) deletion in the third exon. Seven homologous PES2 genes including BnaA02.PES2-2 (BnaA02g28340D) and BnaC02.PES2-2 (BnaC02g36410D) were identified in B. napus (2n = 38, AACC), an allotetraploid derived from B. rapa and B. oleracea (2n = 18, CC). Knockout mutants of either one or two of BnaA02.PES2-2 and BnaC02.PES2-2 in the yellow-flowered B. napus cv. Westar by the CRISPR/Cas9 system showed pale-yellow or white flowers. The knock-out mutants of BnaA02.PES2-2 and BnaC02.PES2-2 had fewer esterified carotenoids. These results demonstrated that BraA02.PES2-2 in B. rapa, and BnaA02.PES2-2 and BnaC02.PES2-2 in B. napus play important roles in carotenoids esterification in chromoplasts that contributes to the accumulation of carotenoids in flower petals.

Genome-wide detection of genotype environment interactions for flowering time in Brassica napus.

Flowering time is strongly related to the environment, while the genotype-by-environment interaction study for flowering time is lacking in Brassica napus. Here, a total of 11,700,689 single nucleotide polymorphisms in 490 B. napus accessions were used to associate with the flowering time and related climatic index in eight environments using a compressed variance-component mixed model, 3VmrMLM. As a result, 19 stable main-effect quantitative trait nucleotides (QTNs) and 32 QTN-by-environment interactions (QEIs) for flowering time were detected. Four windows of daily average temperature and precipitation were found to be climatic factors highly correlated with flowering time. Ten main-effect QTNs were found to be associated with these flowering-time-related climatic indexes. Using differentially expressed gene (DEG) analysis in semi-winter and spring oilseed rapes, 5,850 and 5,511 DEGs were found to be significantly expressed before and after vernalization. Twelve and 14 DEGs, including 7 and 9 known homologs in Arabidopsis, were found to be candidate genes for stable QTNs and QEIs for flowering time, respectively. Five DEGs were found to be candidate genes for main-effect QTNs for flowering-time-related climatic index. These candidate genes, such as BnaFLCs, BnaFTs, BnaA02.VIN3, and BnaC09.PRR7, were further validated by the haplotype, selective sweep, and co-expression networks analysis. The candidate genes identified in this study will be helpful to breed B. napus varieties adapted to particular environments with optimized flowering time.

Eight high-quality genomes reveal pan-genome architecture and ecotype differentiation of Brassica napus.

Rapeseed (Brassica napus) is the second most important oilseed crop in the world but the genetic diversity underlying its massive phenotypic variations remains largely unexplored. Here, we report the sequencing, de novo assembly and annotation of eight B. napus accessions. Using pan-genome comparative analysis, millions of small variations and 77.2-149.6 megabase presence and absence variations (PAVs) were identified. More than 9.4% of the genes contained large-effect mutations or structural variations. PAV-based genome-wide association study (PAV-GWAS) directly identified causal structural variations for silique length, seed weight and flowering time in a nested association mapping population with ZS11 (reference line) as the donor, which were not detected by single-nucleotide polymorphisms-based GWAS (SNP-GWAS), demonstrating that PAV-GWAS was complementary to SNP-GWAS in identifying associations to traits. Further analysis showed that PAVs in three FLOWERING LOCUS C genes were closely related to flowering time and ecotype differentiation. This study provides resources to support a better understanding of the genome architecture and acceleration of the genetic improvement of B. napus.

Identification and characterization of a new dwarf locus DS-4 encoding an Aux/IAA7 protein in Brassica napus.

KEY MESSAGE: A dominant dwarfing gene, ds - 4 , encodes an Aux/IAA protein that negatively regulates plant stature through an auxin signaling pathway. Dwarfism is an important agronomic trait affecting yield in many crop species. The molecular mechanisms underlying dwarfism in oilseed rape (Brassica napus) are poorly understood, restricting the progress of breeding dwarf varieties in this species. Here, we identified and characterized a new dwarf locus, DS-4, in B. napus. Next-generation sequencing-assisted genetic mapping and candidate gene analysis found that DS-4 encodes a nucleus-targeted auxin/indole-3-acetic acid (Aux/IAA) protein. A substitution (P87L) was found in the highly conserved degron motif of the Aux/IAA7 protein in the ds-4 mutant. This mutation co-segregated with the phenotype of individuals in the BC1F2 population. The P87L substitution was confirmed as the cause of the extreme dwarf phenotype by ectopic expression of the mutant allele BnaC05.iaa7 (equivalent to ds-4) in Arabidopsis. The P87L substitution blocked the interaction of BnaC05.iaa7 with TRANSPORT INHIBITOR RESPONSE 1 in the presence of auxin. The BnaC05.IAA7 gene is highly expressed in the cotyledons, hypocotyls, stems and leaves, but weakly in the roots and seeds of B. napus. Our findings provide new insights into the molecular mechanisms underlying dominant (gain-of-function) dwarfism in B. napus. Our identification of a distinct gene locus controlling plant height may help to improve lodging resistance in oilseed rape.

A CACTA-like transposable element in the upstream region of BnaA9.CYP78A9 acts as an enhancer to increase silique length and seed weight in rapeseed.

Rapeseed (Brassica napus L.) is a model plant for polyploid crop research and the second-leading source of vegetable oil worldwide. Silique length (SL) and seed weight are two important yield-influencing traits in rapeseed. Using map-based cloning, we isolated qSLWA9, which encodes a P450 monooxygenase (BnaA9.CYP78A9) and functions as a positive regulator of SL. The expression level of BnaA9.CYP78A9 in silique valves of the long-silique variety is much higher than that in the regular-silique variety, which results in elongated cells and a prolonged phase of silique elongation. Plants of the long-silique variety and transgenic plants with high expression of BnaA9.CYP78A9 had a higher concentration of auxin in the developing silique; this induced a number of auxin-related genes but no genes in well-known auxin biosynthesis pathways, suggesting that BnaA9.CYP78A9 may influence auxin concentration by affecting auxin metabolism or an unknown auxin biosynthesis pathway. A 3.7-kb CACTA-like transposable element (TE) inserted in the 3.9-kb upstream regulatory sequence of BnaA9.CYP78A9 elevates the expression level, suggesting that the CACTA-like TE acts as an enhancer to stimulate high gene expression and silique elongation. Marker and sequence analysis revealed that the TE in B. napus had recently been introgressed from Brassica rapa by interspecific hybridization. The insertion of the TE is consistently associated with long siliques and large seeds in both B. napus and B. rapa collections. However, the frequency of the CACTA-like TE in rapeseed varieties is still very low, suggesting that this allele has not been widely used in rapeseed breeding programs and would be invaluable for yield improvement in rapeseed breeding.