Genetic analysis of global faba bean diversity, agronomic traits and selection signatures.

KEY MESSAGE: We identified marker-trait associations for key faba bean agronomic traits and genomic signatures of selection within a global germplasm collection. Faba bean (Vicia faba L.) is a high-protein grain legume crop with great potential for sustainable protein production. However, little is known about the genetics underlying trait diversity. In this study, we used 21,345 high-quality SNP markers to genetically characterize 2678 faba bean genotypes. We performed genome-wide association studies of key agronomic traits using a seven-parent-MAGIC population and detected 238 significant marker-trait associations linked to 12 traits of agronomic importance. Sixty-five of these were stable across multiple environments. Using a non-redundant diversity panel of 685 accessions from 52 countries, we identified three subpopulations differentiated by geographical origin and 33 genomic regions subjected to strong diversifying selection between subpopulations. We found that SNP markers associated with the differentiation of northern and southern accessions explained a significant proportion of agronomic trait variance in the seven-parent-MAGIC population, suggesting that some of these traits were targets of selection during breeding. Our findings point to genomic regions associated with important agronomic traits and selection, facilitating faba bean genomics-based breeding.

Identification and genetic characterization by high-throughput SNP analysis of intervarietal substitution lines of rapeseed (Brassica napus L.) with enhanced embryogenic potential.

KEY MESSAGE: Seven intervarietal substitution lines were identified with embryogenic potentials up to 40.4 times that of the recurrent parent, providing an ideal material for further in depth studies of this trait. To identify genomic regions that carry genetic factors controlling embryogenic potential of isolated microspores of rapeseed, marker segregations were analysed in a segregating population of haploid microspore-derived embryos and a BC1 population from a cross between 'Express 617' and 'RS239'. After map construction 15 intervarietal substitution lines from the same cross with 'Express 617' as recurrent parent were selected with donor segments covering five genomic regions that had shown skewed segregations in the population of microspore-derived embryos but not in the BC1 population. By comparing the embryogenic potential of microspores of the 15 substitution lines and 'Express 617', seven lines were identified with significantly enhanced embryogenic potential ranging from 4.1 to 40.4 times that of 'Express 617'. To improve the genetic characterization of the selected lines, they were subjected to a high-throughput SNP analysis using the Illumina Infinium 60K chip for rapeseed. Based on 7,960 mapped SNP markers, one to eight donor segments per line, which cover 0.64-6.79% of the 2,126.1 cM of the SNP map, were found. The SNP analysis also gave evidence that homoeologous exchanges had occurred during the development of the substitution line population, increasing the genetic diversity within this population. By comparing donor segments between lines with significantly enhanced embryogenic potential and non-significant lines, 12 genomic regions were identified that may contain genetic factors controlling embryogenic potential in rapeseed. These regions range in size from 0 (represented by just one marker) to 26.8 cM and cover together just 5.42% of the SNP map.

Mapping of QTL for the seed storage proteins cruciferin and napin in a winter oilseed rape doubled haploid population and their inheritance in relation to other seed traits.

KEY MESSAGE: Cruciferin (cru) and napin (nap) were negatively correlated and the cru/nap ratio was closely negative correlated with glucosinolate content indicating a link between the two biosynthetic pathways. Canola-type oilseed rape (Brassica napus L.) is an economically important oilseed crop in temperate zones. Apart from the oil, the canola protein shows potential as a value-added food and nutraceutical ingredient. The two major storage protein groups occurring in oilseed rape are the 2 S napins and 12 S cruciferins. The aim of the present study was to analyse the genetic variation and the inheritance of napin and cruciferin content of the seed protein in the winter oilseed rape doubled haploid population Express 617 × R53 and to determine correlations to other seed traits. Seed samples were obtained from field experiments performed in 2 years at two locations with two replicates in Germany. A previously developed molecular marker map of the DH population was used to map quantitative trait loci (QTL) of the relevant traits. The results indicated highly significant effects of the year and the genotype on napin and cruciferin content as well as on the ratio of cruciferin to napin. Heritabilities were comparatively high with 0.79 for napin and 0.77 for cruciferin. Napin and cruciferin showed a significant negative correlation (-0.36**) and a close negative correlation of the cru/nap ratio to glucosinolate content was observed (-0.81**). Three QTL for napin and two QTL for cruciferin were detected, together explaining 47 and 35 % of the phenotypic variance. A major QTL for glucosinolate content was detected on linkage group N19 whose confidence interval overlapped with QTL for napin and cruciferin content. Results indicate a relationship between seed protein composition and glucosinolate content.

Mapping of QTL for seed dormancy in a winter oilseed rape doubled haploid population.

Following winter oilseed rape cultivation, considerable numbers of volunteer oilseed rape plants may occur in subsequent years in following crops. The appearance of volunteer oilseed rape plants is based on the capability of the seeds to become secondary dormant and to survive in this stage for many years in the soil. Genetic reduction of secondary seed dormancy in oilseed rape could provide a means to reduce the frequency of volunteer plants and especially the dispersal of transgenic oilseed rape. The objective of the present study was to analyse the inheritance of primary and secondary seed dormancy in a winter oilseed rape doubled haploid population derived from the cross Express 617 × R53 and to study correlations to other seed traits. Field experiments were performed in Germany for 2 years at two locations with two replicates. Seeds harvested from open pollinated plants were used for all analyses, including a laboratory test for seed dormancy. A previously developed molecular marker map of the doubled haploid population was used to map QTL of the relevant traits. For primary, secondary and total seed dormancy, the results showed significant effects of the genotypes and their interactions, with years and locations. Two, four and five QTL were detected for primary, secondary and total seed dormancy which explained 19, 35 and 42 % of the phenotypic variance, respectively. Results show that secondary seed dormancy is a heritable trait and that selection for low secondary seed dormancy is possible.

Association mapping for phenological, morphological, and quality traits in canola quality winter rapeseed (Brassica napus L.).

QTL mapping by association analysis has recently gained interest in plant breeding research as an alternative to QTL mapping in segregating populations from biparental crosses. In a first experiment on whole-genome association analysis in rapeseed, 684 mapped AFLP markers were tested for association with 14 traits in a set of 84 canola quality winter rapeseed cultivars. For association analysis a general linear model was used. By testing significance of marker-trait associations against a false discovery rate of 0.2, between 1 and 34 associated markers were found for 10 of the 14 traits. Taking into account linkage disequilibrium between the significant markers, these markers represent between 1 and 22 putative QTL for the respective traits. The minimum phenotypic variance explained by the QTL for the different traits ranged from 15% to 53%. A subset of 27 markers were significantly associated with two or more traits. These markers were predominantly shared between traits that were significantly correlated at the phenotypic level. The results show clearly that in rapeseed, QTL mapping by association analysis is a viable alternative to QTL mapping in segregating populations.

Extent and structure of linkage disequilibrium in canola quality winter rapeseed (Brassica napus L.).

Linkage disequilibrium was investigated in canola quality winter rapeseed to analyze (1) the prospects for whole-genome association analyses and (2) the impact of the recent breeding history of rapeseed on linkage disequilibrium. A total of 845 mapped AFLP markers with allele frequencies >or=0.1 were used for the analysis of linkage disequilibrium in a population of 85 canola quality winter rapeseed genotypes. A low overall level of linkage disequilibrium was found with a mean r (2) of only 0.027 over all 356,590 possible marker pairs. At a significance threshold of P = 2.8 x 10(-7), which was derived by a Bonferroni correction from a global alpha-level of 0.1, only 0.78% of the marker pairs were in significant linkage disequilibrium. Among physically linked marker pairs, the level of linkage disequilibrium was about five times higher with more than 10% of marker pairs in significant linkage disequilibrium. Linkage disequilibrium decayed rapidly with distance between linked markers with high levels of linkage disequilibrium extending only for about 2 cM. Owing to the rapid decay of linkage disequilibrium with distance association analyses in canola quality rapeseed will have a significantly higher resolution than QTL analyses in segregating populations by interval mapping, but much larger number of markers will be necessary to cover the whole genome. A major impact of the recent breeding history of rapeseed on linkage disequilibrium could not be observed.

Optical switch based on a fluid-filled photonic crystal fiber Bragg grating.

We report the implementation of an in-fiber optical switch by means of filling a fluid into the air holes of a photonic crystal fiber with a fiber Bragg grating. Such a switch can turn on/off light transmission with an extinction ratio of up to 33 dB within a narrow wavelength range (Bragg wavelength) via a small temperature adjustment of +/-5 degrees C. The switching function is based on the temperature-dependent coupling between the fundamental core mode and the rod modes in the fluid-filled holes resulting from the thermo-optic effect of the filled fluid.

Splicing Ge-doped photonic crystal fibers using commercial fusion splicer with default discharge parameters.

A novel technique for splicing a small core Ge-doped photonic crystal fiber (PCF) was demonstrated using a commercial fusion splicer with default discharge parameters for the splicing of two standard single mode fibers (SMFs). Additional discharge parameter adjustments are not required to splice the PCF to several different SMFs. A low splice loss of 1.0 approximately 1.4 dB is achieved. Low or no light reflection is expected at the splice joint due to the complete fusion of the two fiber ends. The splice joint has a high bending strength and does not break when the bending radius is decreased to 4 mm.

Genetic analysis of heterosis for yield and yield components in rapeseed (Brassica napus L.) by quantitative trait locus mapping.

The main objective in this research was the genetic analysis of heterosis in rapeseed at the QTL level. A linkage map comprising 235 SSR and 144 AFLP markers covering 2045 cM was constructed in a doubled-haploid population from a cross between the cultivar "Express" and the resynthesized line "R53." In field experiments at four locations in Germany 250 doubled-haploid (DH) lines and their corresponding testcrosses with Express were evaluated for grain yield and three yield components. The heterosis ranged from 30% for grain yield to 0.7% for kernel weight. QTL were mapped using three different data sets, allowing the estimation of additive and dominance effects as well as digenic epistatic interactions. In total, 33 QTL were detected, of which 10 showed significant dominance effects. For grain yield, mainly complete dominance or overdominance was observed, whereas the other traits showed mainly partial dominance. A large number of epistatic interactions were detected. It was concluded that epistasis together with all levels of dominance from partial to overdominance is responsible for the expression of heterosis in rapeseed.

QTL for phytosterol and sinapate ester content in Brassica napus L. collocate with the two erucic acid genes.

Improving oil and protein quality for food and feed purposes is an important goal in rapeseed (Brassica napus L.) breeding programs. Rapeseed contains phytosterols, used to enrich food products, and sinapate esters, which are limiting the utilization of rapeseed proteins in the feed industry. Increasing the phytosterol content of oil and lowering sinapate ester content of meal could increase the value of the oilseed rape crop. The objective of the present study was to identify quantitative trait loci (QTL) for phytosterol and sinapate ester content in a winter rapeseed population of 148 doubled haploid lines, previously found to have a large variation for these two traits. This population also segregated for the two erucic acid genes. A close negative correlation was found between erucic acid and phytosterol content (Spearman's rank correlation, r(s) = -0.80**). For total phytosterol content, three QTL were detected, explaining 60% of the genetic variance. The two QTL with the strongest additive effects were mapped on linkage groups N8 and N13 within the confidence intervals of the two erucic acid genes. For sinapate ester content four QTL were detected, explaining 53% of the genetic variance. Again, a close negative correlation was found between erucic acid and sinapate ester content (r(s) = -0.66**) and the QTL with the strongest additive effects mapped on linkage groups N8 and N13 within the confidence intervals of the two erucic acid genes. The results suggests, that there is a pleiotropic effect of the two erucic acid genes on phytosterol and sinapate ester content; the effect of the alleles for low erucic acid content is to increase phytosterol and sinapate ester content. Possible reasons for this are discussed based on known biosynthetic pathways.

Conditional QTL mapping of oil content in rapeseed with respect to protein content and traits related to plant development and grain yield.

Oil content in rapeseed (Brassica napus L.) is generally regarded as a character with high heritability that is negatively correlated with protein content and influenced by plant developmental and yield related traits. To evaluate possible genetic interrelationships between these traits and oil content, QTL for oil content were mapped using data on oil content and on oil content conditioned on the putatively interrelated traits. Phenotypic data were evaluated in a segregating doubled haploid population of 282 lines derived from the F(1) of a cross between the old German cultivar Sollux and the Chinese cultivar Gaoyou. The material was tested at four locations, two each in Germany and in China. QTLMapper version 1.0 was used for mapping unconditional and conditional QTL with additive (a) and locus pairs with additive x additive epistatic (aa) effects. Clear evidence was found for a strong genetic relationship between oil and protein content. Six QTL and nine epistatic locus pairs were found, which had pleiotropic effects on both traits. Nevertheless, two QTL were also identified, which control oil content independent from protein content and which could be used in practical breeding programs to increase oil content without affecting seed protein content. In addition, six additional QTL with small effects were only identified in the conditional mapping. Some evidence was apparent for a genetic interrelationship between oil content and the number of seeds per silique but no evidence was found for a genetic relationship between oil content and flowering time, grain filling period or single seed weight. The results indicate that for closely correlated traits conditional QTL mapping can be used to dissect the genetic interrelationship between two traits at the level of individual QTL. Furthermore, conditional QTL mapping can reveal additional QTL with small effects that are undetectable in unconditional mapping.

QTL of three agronomically important traits and their interactions with environment in a European x Chinese rapeseed population.

A rapeseed population consisted of 282 doubled haploid (DH) lines derived from a cross between a European vality "Sallux" and a Chinese inbred line "Gaoyou" was planted in 4 locations, 2 in Xi'an and Hangzhou, China, and 2 in Goettingen, Germany. Field experiments were carried out to obtain agronomically phenotypic data from above four environments. A linkage map including 125 SSR-markers was constructed and QTL analyses was performed using mixed model approach to detect QTLs showing additive (a), epistasis (aa) as well as their interactions with environments (QE) for three important agronomic traits: plant height, flowering time and maturity. The results demonstrated that each trait was controlled by several QTLs with additive effect and a number of QTLs with epistatic and QE interaction effects. Plant height was controlled by many QTLs (12 loci with a or combined ae, 5 loci with ae). Additive effects were predominant,totally explained 75% of the phenotypic variation and often combined with digenic epistasis. Of 12 main QTLs, 9 showed Gaoyou alleles decreasing plant height. Most of QTLs with QE effects showed ecologically favourable alleles in diverse regions. Five of 7 ae loci showed Gaoyou alleles in Hangzhou and all the ae loci but one had Sollux alleles in two locations of Germany increasing plant height. The digenic epistatic main effect accounted for one third of total additive main effects. In this study,we discovered 7 and 8 loci having significant additive main effects upon flowering time and maturity, respectively. Of them, early flowering and maturity alleles were respectively 6 and 5 derived from Chinese parent Gaoyou. All these QTLs together accounted for around 60% of the phenotypic variation for each trait. Significant ae interactions were detected for flowering time and maturity and parental alleles showed almost evenly dispersal at all environments. Three of 8 main QTLs for maturity were located at similar or identical positions as QTLs for flowering time, which confirmed the close correlation between these two traits. Two QTLs for plant height on linkage groups N14-1 and 19 were located at similar positions as QTL for flowering time and as already known QTLs for oil content. Selection for reduced plant height and early flowering might reduce oil content. Digenic epistatic QTLs both for flowering time and maturity were detected but much less important than QTLs with additive effects.