Diverse regulatory factors associate with flowering time and yield responses in winter-type Brassica napus

Background: Flowering time, plant height and seed yield are strongly influenced by climatic and day-length adaptation in crop plants. To investigate these traits under highly diverse field conditions in the important oilseed crop Brassica napus, we performed a genome-wide association study using data from diverse agroecological environments spanning three continents. Methods: A total of 158 European winter-type B.napus inbred lines were genotyped with 21,623 unique, single-locus single-nucleotide polymorphism (SNP) markers using the Brassica 60 K-SNP Illumina® Infinium consortium array. Phenotypic associations were calculated in the panel over the years 2010–2012 for flowering time, plant height and seed yield in 5 highly diverse locations in Germany, China and Chile, adding up to 11 diverse environments in total. Results: We identified 101 genome regions associating with the onset of flowering, 69 with plant height, 36 with seed yield and 68 cross-trait regions with potential adaptive value. Within these regions, B.napus orthologs for a number of candidate adaptation genes were detected, including central circadian clock components like CIRCADIAN CLOCK- ASSOCIATED 1 (Bna.CCA1) and the important flowering-time regulators FLOWERING LOCUS T (Bna.FT) and FRUITFUL (Bna.FUL). Discussion: Gene ontology (GO) enrichment analysis of candidate regions suggested that selection of genes involved in post-transcriptional and epigenetic regulation of flowering time may play a potential role in adaptation of B. napus to highly divergent environments. The classical flowering time regulators Bna.FLC and Bna.CO were not found among the candidate regions, although both show functional variation. Allelic effects were additive for plant height and yield, but not for flowering time. The scarcity of positive minor alleles for yield in this breeding pool points to a lack of diversity for adaptation that could restrict yield gain in the face of environmental change. Conclusions: Our study provides a valuable framework to further improve the adaptability and yield stability of this recent allopolyploid crop under changing environments. The results suggest that flowering time regulation within an adapted B. napus breeding pool is driven by a high number of small modulating processes rather than major transcription factors like Bna.CO. In contrast, yield regulation appears highly parallel, therefore yield could be increased by pyramiding positively associated haplotypes

Linkage disequilibrium in the genome of synthetic Brassica napus populations

New technologies as high-density single-nucleotide polymorphism (SNP) genotyping arrays are a powerful tool that can give valuable insight into patterns of linkage disequilibrium (LD) in the recent domesticated Brassica napus genome. This study used the Brassica 60k SNP Illumina consortium genotyping array to assess the distribution of LD and haplotype structure in a diverse panel of 200 synthetic lines of winter oilseed rape (Brassica napus). Pairwise LD analysis was conducted within the A- and C-subgenomes of oilseed rape. Results revealed that LD decayed, on average, more rapidly in the A-subgenome (0.15 Mb) than in the C-subgenome (2.00 Mb). Our findings suggest the presence of a strong selection of large genomic regions associated with important agronomical traits, especially on the Csubgenome. These results imply that during oilseed rape artificial and natural selection, the C-subgenome was of particular interest for breeders. Increasing the genetic diversity and recombination rate on the whole genome level is of crucial importance for future breeding efforts

Genetic structure of synthetic Brassica napus L. Populations

The crop species Brassica napus L. has significant economic importance around the world. However, the complex evolutionary history and vast geographical distribution of oilseed rape has contributed vastly to genetic population structure investigations. Constant breeding efforts, for use for oilseed rape as food for human consumption, and fodder for livestock, have generated new phenotypic diversity. In this study, we used crosses among very diverse morphotypes as Brassica oleracea (turnip rape), conv. capitata var. medullosa (Cavalier rouge), conv. capitata var. sabauda (Savoy 'Wirsing'), conv. botrytis var. alboglabra (broccoli); Brassica rapa (turnip), var. trilocularis (yellow sarson), var. chinensis (bok choy); Brassica cretica; Brassica montana. Until now, genetic studies had insufficient genotypes to determine the relationship of oilseed morphotypes and their genetic population structure. We used 18,272 single nucleotide polymorphism markers in a synthetic nested association mapping diverse panel of 200 B. napus accessions that included crosses of five very diverse parental lines and a common elite accession. Results on population genetic structure and phylogenetic analyses revealed, as expected, five subpopulations that were largely reflective of phenotypes. The results of this study have provided improved resolution to the genetic and phylogenetic relationships of a synthetic panel within the Brassicas species. Understanding genetic diversity available is key to the future genetic study and constant improvement of this important agronomical crop species

Capturing sequence variation among flowering-time regulatory gene homologs in the allopolyploid crop species Brassica napus

Flowering, the transition from the vegetative to the generative phase, is a decisive time point in the lifecycle of a plant. Flowering is controlled by a complex network of transcription factors, photoreceptors, enzymes and miRNAs. In recent years, several studies gave rise to the hypothesis that this network is also strongly involved in the regulation of other important lifecycle processes ranging from germination and seed development through to fundamental developmental and yield-related traits. In the allopolyploid crop species Brassica napus, (genome AACC), homoeologous copies of flowering time regulatory genes are implicated in major phenological variation within the species, however the extent and control of intraspecific and intergenomic variation among flowering-time regulators is still unclear. To investigate differences among B. napus morphotypes in relation to flowering-time gene variation, we performed targeted deep sequencing of 29 regulatory flowering-time genes in four genetically and phenologically diverse B. napus accessions. The genotype panel included a winter-type oilseed rape, a winter fodder rape, a spring-type oilseed rape (all B. napus ssp. napus) and a swede (B. napus ssp. napobrassica), which show extreme differences in winter-hardiness, vernalization requirement and flowering behaviour. A broad range of genetic variation was detected in the targeted genes for the different morphotypes, including non-synonymous SNPs, copy number variation and presence-absence variation. The results suggest that this broad variation in vernalisation, clock and signaling genes could be a key driver of morphological differentiation for flowering-related traits in this recent allopolyploid crop species

Genome-wide association studies in a barley (Hordeum vulgare) diversity set reveal a limited number of loci for resistance to spot blotch (Bipolaris sorokiniana)

Abstract Spot blotch caused by Bipolaris sorokiniana is an important disease in barley worldwide, causing considerable yield losses and reduced grain quality. In order to identify QTL conferring resistance to spot blotch, a highly diverse worldwide barley set comprising 449 accessions was phenotyped for seedling resistance with three isolates (No 31, SH 15 and SB 61) and for adult plant resistance at two locations (Russia and Australia) in two years. Genotyping with the 50 k iSelect barley SNP genotyping chip yielded 33,818 informative markers. Genome-wide association studies (GWAS) using a compressed mixed linear model, including population structure and kinship, revealed 38 significant marker-trait associations (MTA) for spot blotch resistance. The MTA corresponded to two major QTL on chromosomes 1H and 7H and a putative new minor QTL on chromosome 7H explaining between 2.79% and 13.67% of the phenotypic variance. A total of 10 and 14 high-confidence genes were identified in the respective major QTL regions, seven of which have a predicted involvement in pathogen recognition or defence

Quantitative genetic analysis of phenolic acids in oilseed rape meal

Rapeseed meal, a by-product of oilseed extraction related to the agri-food and biofuel industries due to its favourable composition of essential amino acids, is currently utilised for animal feed. In this study, 166 doubled haploid (DH) rapeseed lines from the segregating Brassica napus doubled haploid population YE2-DH were genetically and chemically analysed for phenolic acids. The relationship between seed colour and phenolic acid fractions in B. napuswas investigated using these analyses to improve the quality of rapeseed meal. High-performance liquid chromatography assays were used to estimate phenolic acid content, and the outcomes were used to identify quantitative trait loci (QTL). Nine quantitative feature loci for three distinct phenolic acid compounds were mapped to seven linkage groups. A minor QTL for sinapine was located on linkage group C05 in the same interval as the QTL for seed colour. On chromosome A09, two loci for phenolic acids colocalised with the main QTL for seed colour. Closely linked molecular markers for the target traits (seed colour, phenolic acids) identified during this study could be useful tools for breeding new oilseed rape cultivars with lower levels of antinutritive compounds

Haplotype blocks for genomic prediction: a comparative evaluation in multiple crop datasets

In modern plant breeding, genomic selection is becoming the gold standard for selection of superior genotypes. The basis for genomic prediction models is a set of phenotyped lines along with their genotypic profile. With high marker density and linkage disequilibrium (LD) between markers, genotype data in breeding populations tends to exhibit considerable redundancy. Therefore, interest is growing in the use of haplotype blocks to overcome redundancy by summarizing co-inherited features. Moreover, haplotype blocks can help to capture local epistasis caused by interacting loci. Here, we compared genomic prediction methods that either used single SNPs or haplotype blocks with regards to their prediction accuracy for important traits in crop datasets. We used four published datasets from canola, maize, wheat and soybean. Different approaches to construct haplotype blocks were compared, including blocks based on LD, physical distance, number of adjacent markers and the algorithms implemented in the software “Haploview” and “HaploBlocker”. The tested prediction methods included Genomic Best Linear Unbiased Prediction (GBLUP), Extended GBLUP to account for additive by additive epistasis (EGBLUP), Bayesian LASSO and Reproducing Kernel Hilbert Space (RKHS) regression. We found improved prediction accuracy in some traits when using haplotype blocks compared to SNP-based predictions, however the magnitude of improvement was very trait- and model-specific. Especially in settings with low marker density, haplotype blocks can improve genomic prediction accuracy. In most cases, physically large haplotype blocks yielded a strong decrease in prediction accuracy. Especially when prediction accuracy varies greatly across different prediction models, prediction based on haplotype blocks can improve prediction accuracy of underperforming models. However, there is no “best” method to build haplotype blocks, since prediction accuracy varied considerably across methods and traits. Hence, criteria used to define haplotype blocks should not be viewed as fixed biological parameters, but rather as hyperparameters that need to be adjusted for every dataset