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Meiotic recombination is crucial for assuring proper segregation of parental chromosomes and generation of novel allelic combinations. As this process is tightly regulated, identifying factors influencing rate, and distribution of meiotic crossovers (COs) is of major importance, notably for plant breeding programs. However, high-resolution recombination maps are sparse in most crops including the Brassica genus and knowledge about intraspecific variation and sex differences is lacking. Here, we report fine-scale resolution recombination landscapes for 10 female and 10 male crosses in Brassica oleracea, by analyzing progenies of five large four-way-cross populations from two reciprocally crossed F1s per population. Parents are highly diverse inbred lines representing major crops, including broccoli, cauliflower, cabbage, kohlrabi, and kale. We produced approximately 4.56T Illumina data from 1248 progenies and identified 15 353 CO across the 10 reciprocal crosses, 51.13% of which being mapped to

Background: Arabidopsis thaliana leaf adaxial-abaxial (ad-abaxial) polarity affects leaf morphology and function. Several genes are known to govern ad-abaxial patterning, but the genetic machinery underlying this process is unclear. Results: To uncover critical genes involved in leaf ad-abaxial patterning, we applied a combination of in silico prediction using machine learning (ML) and experimental analysis. Genes known to influence ad-abaxial polarity were utilized as ground truth to train a Random Forest model. This model used gene expression data from various tissues and conditions and promoter regulation data (based on ChIP-seq data) as input. This approach enabled us to identify novel ad-abaxial polarity-related genes. Parallel to this, available and newly obtained transcriptome data enabled us to identify genes differentially expressed across leaf ad-abaxial sides. The integration of the set of differentially expressed genes with the ML model predictions validated the model and led to our final set of predicted leaf polarity influencing genes. Conclusions: Using a combination of in silico prediction using ML and experimental analysis, we obtained a strategy to identify which genes are involved in leaf ad-abaxial specialization. Our final set of 111 genes expands the knowledge of genetic components underlying leaf ad-abaxial polarity specification.

Key message Correlations between morphological traits of cabbage rosette leaves and heads were found. Genome-wide association studies of these traits identified 50 robust quantitative trait loci in multiple years. Half of these loci affect both organs. Abstract Cabbage (Brassica oleracea var. capitata) is an economically important vegetable crop cultivated worldwide. Cabbage plants go through four vegetative stages: seedling, rosette, folding and heading. Rosette leaves are the largest leaves of cabbage plants and provide most of the energy needed to produce the leafy head. To understand the relationship and the genetic basis of leaf development and leafy head formation, 308 cabbage accessions were scored for rosette leaf and head traits in three-year field trials. Significant correlations were found between morphological traits of rosette leaves and heads, namely leaf area with the head area, height and width, and leaf width with the head area and head height, when heads were harvested at a fixed number of days after sowing. Fifty robust quantitative trait loci (QTLs) for rosette leaf and head traits distributed over all nine chromosomes were identified with genome-wide association studies. All these 50 loci were identified in multiple years and generally affect multiple traits. Twenty-five of the QTL were associated with both rosette leaf and leafy head traits. We discuss thirteen candidate genes identified in these QTL that are expressed in heading leaves, with an annotation related to auxin and other phytohormones, leaf development, and leaf polarity that likely play a role in leafy head development or rosette leaf expansion.

Brassica oleraceais an economically important vegetable and fodder crop species that includes many morphotypes exhibiting enormous phenotypic variations. Previously, a pan-genome study based on short reads mapping approach has shown extensive structural variations betweenB. oleraceamorphotypes. Here, to capture more complete genome sequences ofB. oleracea, we report new chromosome-scale genome assemblies for five different morphotypes, namely broccoli, cauliflower, kale, kohlrabi and white cabbage, which were created by combining long-read sequencing data and Bionano DLS optical maps. The five assemblies are the most continuous and completeB. oleraceagenomes to date (contig N50 > 10 Mb). Comparative analysis revealed both highly syntenic relationships and extensive structural variants among the five genomes. Dispensable and specific gene clusters accounted for ~38.19% of total gene clusters based on a pan-genome analysis including our five newly assembled genomes and four previously reported genomes. Using the pan-genome ofB. oleraceaandB. rapa, we revealed their different evolutionary dynamics of LTR-RTs. Furthermore, we inferred the ancestral genome ofB. oleraceaand the common ancestral genome ofB. oleraceaandB. rapavia a pan-genome approach. We observed faster WGT-derived gene loss inB. rapathan inB. oleraceabefore intraspecific diversification. We also revealed continuing gene loss bias during intraspecific diversification of the two species and a strong bias towards losing only one copy among the three paralogous genes. This study provides valuable genomic resources forB. oleraceaimprovement and insights towards understanding genome evolution during the intraspecific diversification ofB. oleraceaandB. rapa.

Background Cabbage white butterflies (Pieris spp.) can be severe pests of Brassica crops such as Chinese cabbage, Pak choi (Brassica rapa) or cabbages (B. oleracea). Eggs of Pieris spp. can induce a hypersensitive response-like (HR-like) cell death which reduces egg survival in the wild black mustard (B. nigra). Unravelling the genetic basis of this egg-killing trait in Brassica crops could improve crop resistance to herbivory, reducing major crop losses and pesticides use. Here we investigated the genetic architecture of a HR-like cell death induced by P. brassicae eggs in B. rapa. Results A germplasm screening of 56 B. rapa accessions, representing the genetic and geographical diversity of a B. rapa core collection, showed phenotypic variation for cell death. An image-based phenotyping protocol was developed to accurately measure size of HR-like cell death and was then used to identify two accessions that consistently showed weak (R-o-18) or strong cell death response (L58). Screening of 160 RILs derived from these two accessions resulted in three novel QTLs for Pieris brassicae-induced cell death on chromosomes A02 (Pbc1), A03 (Pbc2), and A06 (Pbc3). The three QTLs Pbc1–3 contain cell surface receptors, intracellular receptors and other genes involved in plant immunity processes, such as ROS accumulation and cell death formation. Synteny analysis with A. thaliana suggested that Pbc1 and Pbc2 are novel QTLs associated with this trait, while Pbc3 also contains an ortholog of LecRK-I.1, a gene of A. thaliana previously associated with cell death induced by a P. brassicae egg extract. Conclusions This study provides the first genomic regions associated with the Pieris egg-induced HR-like cell death in a Brassica crop species. It is a step closer towards unravelling the genetic basis of an egg-killing crop resistance trait, paving the way for breeders to further fine-map and validate candidate genes.

Brassica oleracea displays enormous phenotypic variation, including vegetables like cabbage, broccoli, cauliflower, kohlrabi, kales etc. Its domestication has not been clarified, despite several genetic studies and investigations of ancient literature. We used 14 152 high-quality SNP markers for population genetic studies and species-tree estimation (treating morphotypes as “species”) using SVD-quartets coalescent-modelling of a collection of 912 globally distributed accessions representing ten morphotypes of B. oleracea, wild B. oleracea accessions and nine related C9 Brassica species. Our genealogical tree provided evidence for two domestication lineages, the “leafy head” lineage (LHL) and the “arrested inflorescence” lineage (AIL). It also showed that kales are polyphyletic with regards to B. oleracea morphotypes, which fits ancient literature describing highly diverse kale types at around 400 BC. The SVD-quartets species tree topology showed that different kale clades are sister to either the LHL or the AIL. Cabbages from the middle-east formed the first-branching cabbage-clade, supporting the hypothesis that cabbage domestication started in the middle-east, which is confirmed by archeological evidence and historic writings. We hypothesize that cabbages and cauliflowers stem from kales introduced from Western Europe to the middle-east, possibly transported with the tin-trade routes in the Bronze age, to be re-introduced later into Europe. Cauliflower is the least diverse morphotype showing strong genetic differentiation with other morphotypes except broccoli, suggesting a strong genetic bottleneck. Genetic diversity reduced from landraces to modern hybrids for almost all morphotypes. This comprehensive Brassica C-group germplasm collection provides valuable genetic resources and a sound basis for B. oleracea breeding.

The Brassica rapa (B. rapa) species displays enormous phenotypic diversity, with leafy vegetables, storage root vegetables and oil crops. These different crops all have different flowering time, which determine their growing season and cultivation area. Little is known about the effects of diverse temperature and day-lengths on flowering time QTL associated with FLC paralogues. We phenotyped the flowering time of a doubled haploid population, established from a cross between Yellow sarson and Pak choi under diverse environmental conditions. We identified flowering-time QTL (fQTL) in different photoperiod and temperature regimes in the greenhouse, and studied their colocation with known flowering time genes. As several fQTL colocalized with FLC paralogues, we studied the expression patterns of four FLC paralogues during the course of vernalization in parental lines. Under all environmental conditions tested the major fQTL that mapped to the BrFLC2_A02 locus was detected, however its effect decreased when plants were grown at low temperatures. Another fQTL that mapped to the FLC paralogue, BrFLC5_A03 was also identified under all tested environments, while no fQTL colocated with BrFLC1_A10 or BrFLC3_A03. Furthermore, the vernalization treatment decreased expression of all BrFLC paralogues in the parental lines, and showed the lowest transcript level after 28 days of vernalization. Transcript abundance stayed low after returning the plants for seven days to normal growth temperature. Interestingly, transcript abundance of BrFLC3_A03 and BrFLC5_A03 was repressed much stronger and already reached lowest levels after 14d in the early-flowering type YS-143. This study improves understanding of the effects of daylength and vernalization on flowering time in B. rapa and the role of the different BrFLC paralogues therein.

A high-quality reference genome is critical for understanding genome structure, genetic variation, and evolution of a species. Of the three available Brassica oleracea genomes, cabbage ‘Cap02-12’ and kale-like rapid-cycling ‘TO1000’ were sequenced and assembled by short reads, hindering assembly through complex repeat regions, while broccoli ‘HDEM’ was assembled using Nanopore long reads combined with optical mapping. In this chapter we present the de novo genome assembly of cauliflower inbred line ‘Korso’ using a strategy combining PacBio long reads, optical mapping, and chromosome-contact maps (Hi-C). The assembly and annotation results of cauliflower ‘Korso’ are compared with the different genome assemblies published before. B. oleracea genome assemblies of cauliflower and broccoli, generated by new long-range technologies (PacBio and Nanopore) were longer, completer, and more continuous, with fewer errors, with improved annotation protein-coding genes, transposon distributions, and centromere positions and sequences. Compared to all other three assemblies, more repeat elements were predicted in ‘Korso’, and the arrangement of the genomic blocks representing the three subgenomes was improved. The availability of new morphotype specific high-quality reference genomes offers an unprecedented opportunity for extensive comparison of intraspecific genome diversity in B. oleracea.

Background Brassica oleracea includes several morphologically diverse, economically important vegetable crops, such as the cauliflower and cabbage. However, genetic variants, especially large structural variants (SVs), that underlie the extreme morphological diversity of B. oleracea remain largely unexplored. Results Here we present high-quality chromosome-scale genome assemblies for two B. oleracea morphotypes, cauliflower and cabbage. Direct comparison of these two assemblies identifies ~ 120 K high-confidence SVs. Population analysis of 271 B. oleracea accessions using these SVs clearly separates different morphotypes, suggesting the association of SVs with B. oleracea intraspecific divergence. Genes affected by SVs selected between cauliflower and cabbage are enriched with functions related to response to stress and stimulus and meristem and flower development. Furthermore, genes affected by selected SVs and involved in the switch from vegetative to generative growth that defines curd initiation, inflorescence meristem proliferation for curd formation, maintenance and enlargement, are identified, providing insights into the regulatory network of curd development. Conclusions This study reveals the important roles of SVs in diversification of different morphotypes of B. oleracea, and the newly assembled genomes and the SVs provide rich resources for future research and breeding.

Additional file 1: Figure S1. Morphology of cauliflower Koroso_1401 and pointed cabbage OX-heart_923. Figure S2. Genome size estimation using flow cytometry analysis for cauliflower Korso and cabbage OX-heart. Figure S3. Chromosome conformation capture of cauliflower Korso and pointed cabbage OX-heart. Figure S4. Pairwise comparisons of the pseudo-chromosomes among cauliflower Korso, cabbage OX-heart and broccoli HDEM. Figure S5. Distribution of estimated insertion times of intact LTR retrotransposons in Korso, OX-heart and B. rapa (v3.0) genomes. Figure S6. Distribution of centromeric satellite repeats CentBr 1 and CentBr 2 on the nine chromosomes of Korso and OX-heart. Figure S7. Segmental collinearity between the A. thaliana genome and genomes of Korso, OX-heart and B. rapa. Figure S8. Subgenomes in Korso and OX-heart. Figure S9. Detected SVs between the genomes of Korso and the OX-heart. Figure S10. Significantly enriched GO terms in genes overlapping with detected SVs in their gene body and/or promoter regions. Figure S11. Distribution of SV genotyping rates in 271 B. oleracea accessions. Figure S12. Population analysis of the 271 B. oleracea accessions using SNPs. Figure S13. Manhattan plot displaying the -log(q-value) of allele variations between cabbage and cauliflower for SVs across the Korso genome. Figure S14. Expression profiles of genes potentially involved in curd maintenance and floral arrest.

The agricultural and consumer quality of Chinese cabbage is determined by its shape. The shape is defined by the folding of the heading leaves, which defines the head top shape (HTS). The overlapping HTS, in which the heading leaves curve inward and overlap at the top, is the shape preferred by consumers. To understand the genetic regulation of HTS, we generated a large segregating F2 population from a cross between pak choi and Chinese cabbage, with phenotypes ranging from nonheading to heading with either outward curving or inward curving overlapping heading leaves. HTS was correlated with plant height, outer/rosette leaf length, and petiole length. A high-density genetic map was constructed. Quantitative trait locus (QTL) analysis resulted in the identification of 22 QTLs for leafy head-related traits, which included five HTS QTLs. Bulked segregant analysis (BSA) was used to confirm HTS QTLs and identify candidate genes based on informative single-nucleotide polymorphisms. Interestingly, the HTS QTLs colocalized with QTLs for plant height, outer/rosette leaf, and petiole length, consistent with the observed phenotypic correlations. Combined QTL analysis and BSA laid a foundation for molecular marker-assisted breeding of Chinese cabbage HTS and directions for further research on the genetic regulation of this trait.

Turnip (Brassica rapa spp. rapa) is an important vegetable species, with a unique physiology. Several plant parts, including both the turnip tubers and leaves, are important for human consumption. During the development of turnip plants, the leaves function as metabolic source tissues, while the tuber first functions as a sink, while later the tuber turns into a source for development of flowers and seeds. In the present study, chemical changes were determined for two genotypes with different genetic background, and included seedling, young leaves, mature leaves, tuber surface, tuber core, stalk, flower and seed tissues, at seven different time points during plant development. As a basis for understanding changes in glucosinolates during plant development, the profile of glucosinolates was analysed using liquid chromatography (LC) coupled to mass spectrometry (MS). This analysis was complemented by a gene expression analysis, focussed on GLS biosynthesis, which could explain part of the observed variation, pointing to important roles of specific gene orthologues for defining the chemical differences. Substantial differences in glucosinolate profiles were observed between above-ground tissues and turnip tuber, reflecting the differences in physiological role. In addition, differences between the two genotypes and between tissues that were harvested early or late during the plant lifecycle. The importance of the observed differences in glucosinolate profile for the ecophysiology of the turnip and for breeding turnips with optimal chemical profiles is discussed.

Chinese cabbage plants go through seedling and rosette stages before forming their leafy head. Chinese cabbage plants resemble pak-choi plants at their seedling stage, but in their rosette stage the leaves of Chinese cabbage differentiate, as they increase in size with shorter petioles. In order to understand the molecular pathways that play a role in leafy head formation, transcript abundance of young emerging leaves was profiled during development of two Chinese cabbage genotypes and a single pak-choi genotype. The two Chinese cabbages differed in many aspects, among others earliness, leaf size and shape, leaf numbers, and leafy head shape. Genome-wide transcriptome analysis clearly separated the seedling stages of all three genotypes together with the later stages from pak-choi, from the later developmental stages of both Chinese cabbages (rosette, folding, and heading). Weighted correlation network analysis and hierarchical clustering using Euclidean distances resulted in gene clusters with transcript abundance patterns distinguishing the two Chinese cabbages from pak-choi. Three clusters included genes with transcript abundance affected by both genotype and developmental stage, whereas two clusters showed only genotype effects. This included a genotype by developmental stage cluster highly enriched with the MapMan category photosynthesis, with high expression during rosette and folding in Chinese cabbages and low expression in the heading inner leaves that are not exposed to light. The other clusters contained many genes in the MapMan categories Cell, showing again differences between pak-choi and both Chinese cabbages. We discuss how this relates to the differences in leaf blade growth between Chinese cabbage and pak-choi, especially at the rosette stage. Overall, comparison of the transcriptome between leaves of two very different Chinese cabbages with pak-choi during plant development allowed the identification of specific gene categories associated with leafy head formation.

Brassica species are characterized by their tremendous intraspecific diversity, exemplified by leafy vegetables, oilseeds, and crops with enlarged inflorescences or above ground storage organs. In contrast to potato tubers that are edible storage organs storing energy as starch and are the vegetative propagation modules, the storage organs of turnips, grown from true seed, are swollen hypocotyls with varying degrees of root and stem that mainly store glucose and fructose. To highlight their anatomical origin, we use the term “hypocotyl-tuber” for these turnip vegetative storage organs. We combined cytological, physiological, genetic and transcriptomic approaches, aiming to identify the initial stages, molecular pathways and regulatory genes for hypocotyl-tuber induction in turnips (B. rapa subsp. rapa). We first studied the development of the hypocotyl zone of turnip and Pak choi and found that 16 days after sowing (DAS) morphological changes occurred in the xylem which indicated the early tuberization stage. Tissue culture experiments showed a clear effect of auxin on hypocotyl-tuber growth. Differentially expressed genes between 1 and 6 weeks after sowing in turnip hypocotyls, located in genomic regions involved in tuber initiation and/or tuber growth defined by QTL and selective sweeps for tuber formation, were identified as candidate genes that were studied in more detail for their role in hypocotyl-tuber formation. This included a Bra-FLOR1 paralogue with increased expression 16 DAS, when the hypocotyl starts swelling, suggesting dual roles for duplicated flowering time genes in flowering and hypocotyl-tuber induction. Bra-CYP735A2 was identified for its possible role in tuber growth via trans-zeatin. Weigthed Co-expression Network Analysis (WGCNA) identified 59 modules of co-expressed genes. Bra-FLOR1 and Bra-CYP735A2 were grouped in a module that included several genes involved in carbohydrate transport and metabolism, cell-wall growth, auxin regulation and secondary metabolism that serve as starting points to illuminate the transcriptional regulation of hypocotyl-tuber formation and development., Turnips: “Neeps” show early differences from non-tuber–forming relative Turnips show physiological and molecular signs of hypocotyl-tuber development as early as 16 days after sowing, a finding that could help farmers improve the performance of this important food and feed crop. A team led by Guusje Bonnema from Wageningen University and Research in the Netherlands compared the early anatomy of turnips and another closely related subspecies of Brassica rapa, the non-tuber–forming pak choi. They documented differences in the cellular organization of the xylem by day-16 after seed planting. Gene expression profiling between 1–6 weeks after sowing revealed many genes involved in hypocotyl-tuber initiation and growth. These genes affect a range of biological processes, from carbohydrate transport and metabolism to cell-wall growth to hormone regulation. Tissue culture experiments also showed that auxin, a plant growth hormone, promoted early hypocotyl-tuber development.

Fatty acids in seeds affect seed germination and seedling vigor, and fatty acid composition determines the quality of seed oil. In this study, quantitative trait locus (QTL) mapping of fatty acid and transcript abundance was integrated with gene network analysis to unravel the genetic regulation of seed fatty acid composition in a Brassica rapa doubled haploid population from a cross between a yellow sarson oil type and a black-seeded pak choi. The distribution of major QTLs for fatty acids showed a relationship with the fatty acid types: linkage group A03 for monounsaturated fatty acids, A04 for saturated fatty acids, and A05 for polyunsaturated fatty acids. Using a genetical genomics approach, expression quantitative trait locus (eQTL) hotspots were found at major fatty acid QTLs on linkage groups A03, A04, A05, and A09. An eQTL-guided gene coexpression network of lipid metabolism-related genes showed major hubs at the genes BrPLA2-ALPHA, BrWD-40, a number of seed storage protein genes, and the transcription factor BrMD-2, suggesting essential roles for these genes in lipid metabolism. Three subnetworks were extracted for the economically important and most abundant fatty acids erucic, oleic, linoleic, and linolenic acids. Network analysis, combined with comparison of the genome positions of cis- or trans-eQTLs with fatty acid QTLs, allowed the identification of candidate genes for genetic regulation of these fatty acids. The generated insights in the genetic architecture of fatty acid composition and the underlying complex gene regulatory networks in B. rapa seeds are discussed.

Cauliflower (Brassica oleracea ssp. botrytis) is an important vegetable that is grown worldwide from the tropics to temperate zones. The harvested product is the curd, which consists of arrested inflorescence meristems. The switch from vegetative development to curd formation in cauliflower, referred to as the generative switch, is strongly temperature responsive in the majority of varieties. We aimed at measuring the delay in timing of the generative switch by high ambient temperature, and how temperature affects the expression of genes with a potential role in timing of this switch. A seven day increase of six degrees in day and night temperature during vegetative development, results in a substantial delay of the generative switch and increased variation in timing of this switch in sensitive cultivars only. The expression level of the Cauliflower FRUITFULL-like gene BoFULc increased significantly at the generative switch and therefore can be used as marker for this developmental phase change. The expression profiles of the majority of the other investigated cauliflower flowering time genes resembled the expression behaviour of their homologous genes in the model plant Arabidopsis thaliana during the vegetative stage and flowering induction. An exception was the expression of two FLC paralogues BoFLC-1 and BoFLC-3, which showed opposite expression profiles of which the pattern of BoFLC-1 resembles the pattern expected based on Arabidopsis FLC. This interesting observation suggests different roles for these two FLC paralogs in regulation of the timing of the generative switch in cauliflower. Unexpectedly, high temperatures did not delay timing of expression of the majority of investigated genes in meristems and leaves of sensitive cultivars that were delayed in the switch to the generative stage. However, expression of a few potential flowering-time genes was affected by the high temperature treatment in a sensitive cultivar, making them potential candidates to be causal for the observed delay in generative switch.

Non-heading Chinese cabbage encompasses a wide diversity of morphotypes, like the well-known Pak-choi, Wu ta cai, Cai xin, Tai cai, and Fen nei cai. Despite recent developments in re-sequencing which results in the detection of SNPs, insertions, deletions and copy number variants, there has been no systematical cytogenetic characterization of different non-heading Chinese cabbage morphotypes, aiming to detect large structural variations. To tackle this issue, the karyotypes of five genotypes belonging to five non-heading Chinese cabbage morphotypes were analyzed using fluorescence in situ hybridization (FISH) with ribosomal DNA (rDNA) and blend DAPI band (BDB) on mitotic chromosomes. The ideograms of these five non-heading Chinese cabbage genotypes were produced by exploring and comparing the distributional patterns in terms of rDNA (45S and 5S) and constitutive heterochromatin regions between morphotypes. This study revealed variation in the distributional patterns of ribosomal genes and heterochromatin regions, which point to chromosomal rearrangements in non-heading Chinese cabbage. The 45S rDNA site on chromosome A5 and 5S rDNA signal on chromosome A10 were conserved between all five morphotypes, suggesting that these sites were presented in the ancestor karyotype of non-heading Chinese cabbage. Karyotypic variation and conservation facilitates accurate estimation of the genetic diversity, to better understanding of the evolution, and insight in this variation may facilitate breeding of non-heading Chinese cabbage.

From prokaryotes to eukaryotes, phenotypic variation, adaptation and speciation has been associated with structural variation between genomes of individuals within the same species. Many computer algorithms detecting such variations (callers) have recently been developed, spurred by the advent of the next-generation sequencing technology. Such callers mainly exploit split-read mapping or paired-end read mapping. However, as different callers are geared towards different types of structural variation, there is still no single caller that can be considered a community standard; instead, increasingly the various callers are combined in integrated pipelines. In this article, we review a wide range of callers, discuss challenges in the integration step and present a survey of pipelines used in population genomics studies. Based on our findings, we provide general recommendations on how to set-up such pipelines. Finally, we present an outlook on future challenges in structural variation detection.

The Brassica genus comprises many economically important worldwide cultivated crops. The well-established model of the Brassica genus, U’s triangle, consists of three basic diploid plant species (Brassica rapa, Brassica oleracea and Brassica nigra) and three amphidiploid species (Brassica napus, Brassica juncea and Brassica carinata) that arose through interspecific hybridizations. Despite being extensively studied because of its commercial relevance, several aspects of the origin of the Brassica species and the relationships within and among these six species still remain open questions. Here, we successfully de novo assembled 60 complete chloroplast genomes of Brassica genotypes of all six species. A complete map of the Single Nucleotide Variants (SNVs) and Insertions and Deletions (InDels) in the chloroplast genomes of different Brassica species was produced. The chloroplast genome consists of a Large and a Small Single Copy (LSC and SSC) region between two Inverted Repeats (IRs), and while these regions of chloroplast genomes have very different molecular evolutionary rates, phylogenetic analyses of different regions yielded no contradicting topologies and separated the Brassica genus into four clades. B. carinata and B. juncea share their chloroplast genome with one of their hybridization donors B. nigra and B. rapa, respectively, which fits the U model. B. rapa, surprisingly, shows evidence of two types of chloroplast genomes, with one type specific to some Italian broccoletto accessions. B. napus clearly has evidence for two independent hybridization events, as it contains either B. rapa chloroplast genomes. The divergence estimation suggests that B. nigra and B. carinata diverged from the main Brassica clade 13.7 million years ago (Mya), while B. rapa and B. oleracea diverged at 2.18 Mya. The use of the complete chloroplast DNA sequence not only provides insights into comparative genome analysis but also paves the way for a better understanding of the phylogenetic relationships within the Brassica genus.

The paleohexaploid crop Brassica rapa harbors an enormous reservoir of morphological variation, encompassing leafy vegetables, vegetable and fodder turnips (Brassica rapa, ssp. campestris), and oil crops, with different crops having very different leaf morphologies. In the triplicated B. rapa genome, many genes have multiple paralogs that may be regulated differentially and contribute to phenotypic variation. Using a genetical genomics approach, phenotypic data from a segregating doubled haploid population derived from a cross between cultivar Yellow sarson (oil type) and cultivar Pak choi (vegetable type) were used to identify loci controlling leaf development. Twenty-five colocalized phenotypic quantitative trait loci (QTLs) contributing to natural variation for leaf morphological traits, leaf number, plant architecture, and flowering time were identified. Genetic analysis showed that four colocalized phenotypic QTLs colocalized with flowering time and leaf trait candidate genes, with their cis-expression QTLs and cis- or trans-expression QTLs for homologs of genes playing a role in leaf development in Arabidopsis (Arabidopsis thaliana). The leaf gene Brassica rapa KIP-related protein2_A03 colocalized with QTLs for leaf shape and plant height; Brassica rapa Erecta_A09 colocalized with QTLs for leaf color and leaf shape; Brassica rapa Longifolia1_A10 colocalized with QTLs for leaf size, leaf color, plant branching, and flowering time; while the major flowering time gene, Brassica rapa flowering locus C_A02, colocalized with QTLs explaining variation in flowering time, plant architectural traits, and leaf size. Colocalization of these QTLs points to pleiotropic regulation of leaf development and plant architectural traits in B. rapa.

Thermal processing of Brassica vegetables can lead to substantial loss of potential health-promoting glucosinolates (GLs). The extent of thermal degradation of a specific GL varies in different vegetables, possibly due to differences in the composition of other metabolites within the plant matrices. An untargeted metabolomics approach followed by random forest regression was applied to identify metabolites associated to thermal GL degradation in a segregating Brassica oleracea population. Out of 413 metabolites, 15 were associated with the degradation of glucobrassicin, six with that of glucoraphanin and two with both GLs. Among these 23 metabolites three were identified as flavonols (one kaempferol- and two quercetin-derivatives) and two as other GLs (4-methoxyglucobrassicin, gluconasturtiin). Twenty quantitative trait loci (QTLs) for these metabolites, which were associated with glucoraphanin and glucobrassicin degradation, were identified on linkage groups C01, C07 and C09. Two flavonols mapped on linkage groups C07 and C09 and co-localise with the QTL for GL degradation determined previously.

Nutritional quality of vegetables is affected by several steps in the food chain. Up to now the effects of these different steps are mostly studied separately. We propose the cooperation between plant breeding and food technology by using food technological parameters as breeding traits to identify genetic loci associated with food processing. An example of mechanistic studies of glucosinolate losses during cooking is used to discuss requirements, possibilities, challenges and benefits of such an integrated approach with the final aim to breed for vegetables with higher retention of glucosinolates, as example for other phytochemicals, during food processing.

The closely related species Brassica rapa and B. oleracea encompass a wide range of vegetable, fodder and oil crops. The release of their reference genomes has facilitated resequencing collections of B. rapa and B. oleracea aiming to build their variome datasets. These data can be used to investigate the evolutionary relationships between and within the different species and the domestication of the crops, hereafter named morphotypes. These data can also be used in genetic studies aiming at the identification of genes that influence agronomic traits. We selected and resequenced 199 B. rapa and 119 B. oleracea accessions representing 12 and nine morphotypes, respectively. Based on these resequencing data, we obtained 2,249,473 and 3,852,169 high quality SNPs (single-nucleotide polymorphisms), as well as 303,617 and 417,004 InDels for the B. rapa and B. oleracea populations, respectively. The variome datasets of B. rapa and B. oleracea represent valuable resources to researchers working on evolution, domestication or breeding of Brassica vegetable crops.

Chinese cabbage buds were soaked with Ethyl methanesulfonate (EMS) was used to soak to induce mutagenesis. The influence of different EMS concentrations and treatment durations on microspore development, embryo production rate and seedling rate were evaluated in five genotypes of Chinese cabbage genotypes. Mutations in four color-related genes were identified using hHigh resolution melting (HRM) curves of their PCR products. was used to identify mutations in four color-related genes. The results demonstrated that an EMS concentration greater than 0.1% was lethal to microspores. The greatest embryo production and seedling rates were observed when buds were treated with 0.03 to 0.1% EMS for 5 to 10 min, while EMS concentrations greater than 0.1% were lethal to the microspores.. These conditions also facilitated culturing of microspores. In total, 142 mutants with distinct variations in leaf shape, leaf color, corolla size, flower color, bolting time and downy mildew resistance were identified from 475 microspore culture derived Doubled Haploids plantlets. Our results suggest demonstrate that microspore derived Doubled Haploids from soaking of buds with EMS soaked buds is anrepresents an efficient approach to the rapidly generation generate of Chinese cabbage mutants.