Identification of QTL for oil content,seed yield,and flowering time in oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis>)

A genetic map was constructed with 353 sequence-related amplified polymorphism and 34 simple sequence repeat markers in oilseed rape (Brassica napus L.). The map consists of 19 linkage groups and covers 1,868 cM of the rapeseed genome. A recombinant doubled haploid (DH) population consisting of 150 lines segregating for oil content and other agronomic traits was produced using standard microspore culture techniques. The DH lines were phenotyped for days to flowering, oil content in the seed, and seed yield at three locations for 3 years, generating nine environments. Data from each of the environments were analyzed separately to detect quantitative trait loci (QTL) for these three phenotypic traits. For oil content, 27 QTL were identified on 14 linkage groups; individual QTL for oil content explained 4.20–30.20% of the total phenotypic variance. For seed yield, 18 QTL on 11 linkage groups were identified, and the phenotypic variance for seed yield, as explained by a single locus, ranged from 4.61 to 24.44%. Twenty-two QTL were also detected for days to flowering, and individual loci explained 4.41–48.28% of the total phenotypic variance.     

Analysis of QTL for seed oil content in <Emphasis Type="Italic">Brassica napus</Emphasis> by association mapping and QTL mapping

Increasing seed oil content is one of the most important breeding targets for rapeseed (Brassica napus). In this study, we combined quantitative trait loci (QTL) mapping and marker-trait association analysis to dissect the genetic basis of seed oil content in rapeseed. A doubled haploid (DH) population with 261 lines was grown in two highly contrasting macro-environments, Germany with winter ecotype environment and China with semi-winter ecotype environment, to explore the effect of environment effect of on seed oil content. Notable macro-environment effect was found for seed oil content. 19 QTL for seed oil content were identified across the two macro-environments. For association analysis, a total of 142 rapeseed breeding lines with diverse oil contents were grow in China macro-environment. We identified 23 simple sequence repeat (SSR) markers that were significantly associated with the seed oil content. Comparative analysis revealed that five QTL identified in the DH population, located on chromosomes A03, A09, A10 and C09, were co-localized with 11 significantly associated SSR markers that were identified from the association mapping population. Of which, the QTL on chromosome A10 was found to be homeologous with the QTL on chromosome C09 by aligning QTL confidence intervals with the reference genomes B. napus. Those QTL associated with specific macro-environments provides valuable insight into the genetic regulation of seed oil content and will facilitate marker-assisted breeding of B. napus.     

A SNP genetic linkage map based on the ‘Hamilton’ by ‘Spencer’ recombinant inbred line population identified QTL for seed isoflavone contents in soybean

Soybean is one of the most important crops worldwide for its protein and oil as well as the health beneficial phytoestrogens or isoflavone. This study reports a relatively dense single nucleotide polymorphism (SNP)‐based genetic map based on ‘Hamilton’ by ‘Spencer’ recombinant inbred line population and quantitative trait loci (QTL) for seed isoflavone contents. The genetic map is composed of 1502 SNP markers and covers about 1423.72 cM of the soybean genome. Two QTL for seed isoflavone contents have been identified in this population. One major QTL that controlled both daidzein (qDZ1) and total isoflavone contents (qTI1) was found on LG C2 (Chr 6). And a second QTL for glycitein content (qGT1) was identified on the LG G (Chr 18). These two QTL in addition to others identified in soybean could be used in soybean breeding to optimize isoflavone content. This newly assembled soybean linkage map is a useful tool to identify and map QTL for important agronomic traits and enhance the identification of the genes involved in these traits.     

Quantitative trait loci mapping of seed colour,hairy leaf,seedling anthocyanin,leaf chlorosis and days to flowering in F2 population of Brassica rapa L.

A diversity arrays technology (DArT) map was constructed to identify quantitative trait loci (QTL) affecting seed colour, hairy leaf, seedling anthocyanin, leaf chlorosis and days to flowering in Brassica rapa using a F₂ population from a cross between two parents with contrasting traits. Two genes with dominant epistatic interaction were responsible for seed colour. One major dominant gene controls the hairy leaf trait. Seedling anthocyanin was controlled by a major single dominant gene. The parents did not exhibit leaf chlorosis; however, 32% F₂ plants showed leaf chlorosis in the population. A distorted segregation was observed for days to flowering in the F₂ population. A linkage map was constructed with 376 DArT markers distributed over 12 linkage groups covering 579.7 cM. The DArT markers were assigned on different chromosomes of B. rapa using B. rapa genome sequences and DArT consensus map of B. napus. Two QTL (RSC1‐2 and RSC12‐56) located on chromosome A8 and chromosome A9 were identified for seed colour, which explained 19.4% and 18.2% of the phenotypic variation, respectively. The seed colour marker located in the ortholog to Arabidopsis thaliana Transparent Testa2 (AtTT2). Two QTL RLH6‐0 and RLH9‐16 were identified for hairy leaf, which explained 31.6% and 20.7% phenotypic variation, respectively. A single QTL (RSAn‐12‐157) on chromosome A7, which explained 12.8% of phenotypic variation was detected for seedling anthocyanin. The seedling anthocyanin marker is found within the A. thaliana Transparent Testa12 (AtTT12) ortholog. A QTL (RLC6‐04) for leaf chlorosis was identified, which explained 55.3% of phenotypic variation. QTL for hairy leaf and leaf chlorosis were located 0–4 cM apart on the same chromosome A1. A single QTL (RDF‐10‐0) for days to flowering was identified, which explained 21.4% phenotypic variation.     

Mapping QTL for seed yield and canning quality following processing of black bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.)

Quantitative trait loci (QTL) analysis was conducted to identify QTL for seed yield and color retention following processing of a recombinant inbred line (RIL) black bean population. A population of 96 RILs were derived from the cross of black bean cultivars ‘Jaguar’ and 115M and evaluated in replicated trials at one location over 4 years (2004–2007) in Michigan. A 119-point genetic map constructed using simple sequence repeat (SSR), sequence related amplified polymorphism (SRAP), target region amplified polymorphism (TRAP) and phenotypic markers spanned fifteen linkage groups (LG) or 460 cM of the bean genome. Fourteen QTL for yield and color retention in four environments were identified by composite interval mapping on six linkage groups. A major QTL SY10.2^J115 for seed yield was identified on LG B10 with additional QTL on B3, B5, and B11. Color retention following processing was associated with loci on B1, B3, B5, B8, and B11. 115M possessed positive alleles for yield, but negative alleles for color retention. Some QTL for yield and color retention co-localized with regions identified in previous studies while others, particularly for color retention, were unique. Additional QTL for agronomic and canning quality traits were detected and individual contributions to future black bean breeding are discussed.     

Identification of QTL underlying the resistance of soybean to pod borer, <Emphasis Type="Italic">Leguminivora glycinivorella</Emphasis> (Mats.) obraztsov,and correlations with plant,pod and seed traits

Soybean (Glycine max L. Merr.) pod borer (Leguminivora glycinivorella (Mats.) Obraztsov) (SPB) results in severe loss in soybean yield and quality in certain regions of the world, especially in Northeastern China, Japan and Russia. The aim here was to evaluate the inheritance of pod borer resistance and to identify quantitative trait loci (QTL) underlying SPB resistance for the acceleration of the control of this pest. Used were the 129 recombinant inbred lines (RILs) of the F_5:6 derived population from ‘Dong Nong 1068’ × ‘Dong Nong 8004’ and 131 SSR markers. Correlations between the percentage of damaged seeds (PDS) by pod borer and plant, pod and seed traits that were potentially related to SPB resistance were analyzed. The results showed highly significant correlations between PDS by pod borer and plant height (PH), maturity date (MA), pod color (PC), pubescence density (PB), 100-seed weight (SW) and protein content existed. Soybeans with dwarf stem, light color of pod coat, small seeds, lower density of pubescence, early maturity and low content of protein seemed to have higher resistance to SPB. The correlated traits had potential to inhibit egg deposition and thereby to decrease the damage by SPB. Three QTL directly associated with the resistance to SPB judged by PDS at harvest were identified. qRspb-1 (Satt541–Satt253) and qRspb-2 (Satt253–Satt314) were both on linkage group (LG) H and qRspb-3 (Satt288–Satt199) on LG G. The three QTL explained 10.96, 9.73 and 11.59% of the phenotypic variation for PDS, respectively. In addition, 12 QTL that underlay 10 of 13 traits potentially related with SPB resistance were found. These QTL detected jointly provide potential for marker assisted selection to improve cultivar resistance to SPB. Guiyun Zhao, Jian Wang, and Yingpeng Han have equal contribution to the paper.     

Dissection of a major QTL for seed colour and fibre content in Brassica napus reveals colocalization with candidate genes for phenylpropanoid biosynthesis and flavonoid deposition

A major quantitative trait locus (QTL) influencing seed fibre and colour in Brassica napus was dissected by marker saturation in a doubled haploid (DH) population from the black‐seeded oilseed rape line ‘Express 617’ crossed with a yellow‐seeded B. napus line, ‘1012–98’. The marker at the peak of a sub‐QTL with a strong effect on both seed colour and acid detergent lignin content lay only 4 kb away from a Brassica (H+)‐ATPase gene orthologous to the transparent testa gene AHA10. Near the peak of a second sub‐QTL, we mapped a copy of the key phenylpropanoid biosynthesis gene cinnamyl alcohol dehydrogenase, while another key phenylpropanoid biosynthesis gene, cinnamoyl co‐a reductase 1, was found nearby. In a cross between ‘Express 617’ and another dark‐seeded parent, ‘V8’, Bna.CCR1 was localized in silico near the peak of a corresponding seed fibre QTL, whereas in this case Bna.CAD2/CAD3 lay nearby. Re‐sequencing of the two phenylpropanoid genes via next‐generation amplicon sequencing revealed intragenic rearrangements and functionally relevant allelic variation in the three parents.     

Association of isozyme markers with quantitative trait loci in random single seed descent derived lines of lentil (Lens culinaris Medik.)

Summary Polymorphism at isozyme loci was used to locate factors responsible for variation in quantitative traits of lentil. Eight sets of random single seed descent (RSSD) derived lines were developed by advancing individual F₃ plants of interspecific (L. culinaris Medik. × L. orientalis Boiss.) hybrids to the F₆. The RSSD lines in each of the eight sets differed for alleles at 2–8 isozyme loci. In each set, association of isozyme loci with variation in seven quantitative traits (days to flower, days to mature, plant height, biomass, seed yield, harvest index, seed weight) was determined for each pairwise combination of a quantitative trait with a marker locus. Loci affecting variation in all seven quantitative traits were detected by their association with 14 isozyme markers (Aat-c, Aat-m, Aat-p, Adh-1, Fk, Gal-1, Gal-2, Lap-1, Lap-2, Pgd-p, Pgi, Pgm-c, Pgm-p, Skdh). The known position of 10 the 14 isozyme loci on the lentil genetic map was used to mark the genomic regions for possible location of associated quantitative trait loci (QTL). Detected QTL were found to be located in six of the seven linkage groups on lentil genetic map. Regions of the genome represented by linkage groups, 1, 5 and 7 appeared to affect a greater number of traits than other genomic regions represented by linkage groups 2, 3 and 4. Results indicated that the mean expression of quantitative traits at segregating marker locus classes can be used to locate the genetic factors in lentil which influence the behavior of economically important traits.     

Phenotypic and seed structural comparison in hybrids of different Brassica rapa and B. oleracea

Brassica napus is an important oil species with short history and narrow genetic background. Interspecific hybrids from crosses between B. oleracea and different B. rapa were obtained. We found the hybrids with white petal resembling B. oleracea, the flavonoid and phenolic content decreased in hybrids, agreeing with the expressional changes of flavonoid biosynthesis genes. Seed coat of hybrids resembled diploid parents, or partly resembled to each parent with a clear outline. The palisade layer in hybrids was thicker than parents, with similar pigment accumulation as B. oleracea but more than B. rapa. Differentially sized protein bodies (PBs) were found in hybrids. The radical and inner cotyledon of all hybrids were identified with larger but less PBs than parents. The average size of PBs in outer cotyledon of resynthesized B. napus was also larger than parents, but the number of PBs was not significantly reduced. The phenotypic and seed structural variations after polyploidization of B. napus would be interesting for genetic broadening and breeding of rapeseed.     

Independent selection for seed free tryptophan content and vernalization response in chickpea domestication

Chickpea shows a distinct domestication trajectory vis‐a‐vis pod dehiscence and growth cycle mediated by vernalization insensitivity compared with its companion Near Eastern legumes. Our objectives were: (i) to map the quantitative trait loci (QTLs) associated with vernalization response and seed free tryptophan in domesticated × wild chickpea progeny and (ii) estimate the genetic correlation between vernalization response and free tryptophan content. A domesticated × wild chickpea cross was used to document phenotypic segregation in both traits and to construct a skeletal genetic map for QTL detection. A number of vernalization response and seed free tryptophan content QTLs were documented in both F₂ and F₃ generations. No significant genetic correlation between these two traits was observed. Epistatic relationship between two free tryptophan loci was documented. It is evident that selection for high seed tryptophan is easier to accomplish relative to selection for vernalization insensitivity. This suggests that the two traits were selected independently in antiquity, thereby corroborating earlier claims for conscious selection processes associated with chickpea domestication.     

Association analysis using SSR markers to find QTL for seed protein content in soybean

Association analysis studies can be used to test for associations between molecular markers and quantitative trait loci (QTL). In this study, a genome-wide scan was performed using 150 simple sequence repeat (SSR) markers to identify QTL associated with seed protein content in soybean. The initial mapping population consisted of two subpopulations of 48 germplasm accessions each, with high or low protein levels based on data from the USDA’s Germplasm Resources Information Network website. Intrachromosomal LD extended up to 50 cM with r ² > 0.1 and 10 cM with r ² > 0.2 across the accessions. An association map consisting of 150 markers was constructed on the basis of differences in allele frequency distributions between the two subpopulations. Eleven putative QTL were identified on the basis of highly significant markers. Nine of these are in regions where protein QTL have been mapped, but the genomic regions containing Satt431 on LG J and Satt551 on LG M have not been reported in previous linkage mapping studies. Furthermore, these new putative protein QTL do not map near any QTL known to affect maturity. Since biased population structure was known to exist in the original association analysis population, association analyses were also conducted on two similar but independent confirmation populations. Satt431 and Satt551 were also significant in those analyses. These results suggest that our association analysis approach could be a useful alternative to linkage mapping for the identification of unreported regions of the soybean genome containing putative QTL.     

Molecular markers for yield components in Brassica juncea – do these assist in breeding for high seed yield?

A population of 112 F₁-derived doubled haploid lines was produced from a reciprocal cross of Brassica juncea. The parents differed for seed quality, seed color and many agronomic traits. A detailed RFLP linkage map of this population, comprising 316 loci, had been constructed, and was used to map quantitative trait loci (QTL) for seed yield and yield components, viz. siliqua length, number of seeds per siliqua, number of siliques per main raceme and 1000-seed weight. Stable and significant QTLs were identified for all these yield components except seed yield. For yield components, a selection index based on combined phenotypic and molecular data (QTL effects) could double up the efficiency of selection compared to the expected genetic advance by phenotypic selection. Selection indices for high seed yield, based on the phenotypic data of yield and yield components, could only improve the efficiency of selection by 4% of the genetic advance that can be expected from direct phenotypic selection for yield alone. Inclusion of molecular data together with the phenotypic data of yield components in the selection indices did not improve the efficiency of selection for higher seed yield. This is probably due to often negative relationships among the yield components. Most of the QTLs for yield components were compensating each other, probably due to linkage, pleiotropy or developmentally induced relationships among them. The breeding strategy for B. juncea and challenges to marker assisted selection are discussed.     

QTL identification for molecular breeding of fibre yield and fibre quality traits in jute

In jute (Corchorus olitorius), quantitative trait loci (QTL) analysis was conducted to study the genetics of eight fibre yield traits and two fibre quality traits. For this purpose, we used a mapping population consisting of 120 recombinant inbred lines (RILs) and also used a linkage map consisting of 36 SSR markers that was developed by us earlier (Das et al. 2011). The RIL population was derived from the cross JRO 524 (coarse fibre) × PPO4 (fine fibre) following single seed descent. Using single-locus analysis involving composite interval mapping, a total of 21 QTLs were identified for eight fibre yield traits whereas for fibre quality (fibre fineness), only one QTL was detected. The QTL for fibre fineness explained 8.31–10.56% of the phenotypic variation and was detected in two out of three environments. Using two-locus analysis involving QTLNetwork, as many as 11 M-QTLs were identified for seven fibre yield traits (excluding top diameter) and one M-QTL was identified for fibre fineness which accounted for 4.57% of the phenotypic variation. For six fibre yield traits, we detected 16 E-QTLs involved in nine QQ epistatic interactions. For fibre fineness, four E-QTLs involved in two QQ epistatic interactions and for fibre strength, six E-QTLs involved in three QQ epistatic interactions were identified. Eight out of the 11 M-QTLs observed for the fibre yield traits were also involved in QE interactions; for fibre fineness and fibre strength, no QE interactions were observed.     

Quantitative trait locus analysis of seed germination,seedling vigour and seedling‐regulated hormones in Brassica napus

Good germination and seedling vigour are major breeding targets in winter oilseed rape (Brassica napus), because seedling vigour and prewinter crop establishment are closely associated with postwinter growth and yield. Here, we identified quantitative trait loci (QTL) related to germination, seedling vigour and seedling‐regulated hormones in a doubled haploid (DH) mapping population from a cross between winter oilseed rape parents with high vigour (Express 617) and low vigour (1012‐98). By phenotyping in a climate‐controlled glasshouse, we identified a total of 13 QTL on nine chromosomes for germination and seedling‐related traits at 7 and 14 days after sowing (DAS), explaining up to 11.2% of the phenotypic variation for seedling vigour. Forty‐seven metabolic QTL on 15 chromosomes were identified for auxin, abscisic acid (ABA) and dihydrophaseic acid (DPA) at 5 and 12 DAS, explaining up to 49.4% of phenotypic variation in seedling hormone composition. Multitrait QTL hot spots contribute to our understanding of the genetics and metabolomics of germination and seeding vigour in B. napus, and represent potential targets to breed high‐vigour cultivars.     

Development of linkage map and mapping of QTLs for oil content and yield attributes in linseed (<Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.)

Linseed (Linum usitatissimum L.) is an important oilseed as well as stem fiber crop and rich source of omega-3 fatty acid. The present study aims to develop linkage map based on Indian genotypes and utilize it for mapping QTLs for important agronomic traits. Two diverse parental genotypes (KL-213 and RKY-14) of linseed showed wide range of variability for oil content and yield attributes. These parental genotypes also showed reasonable level of SSR polymorphism (~ 9.0%). The mapping population showed normal distribution of phenotypic traits. One hundred forty-six SSR markers were mapped on 15 linkage groups with marker density ranging from 3 to 18 markers per linkage group at average distance of 14.2 cM. A total of 11 QTLs were identified for six quantitative traits. Three QTLs for capsules/plant, 2 QTLs each for plant height, seeds/capsule and oil content and 1 QTL each for branches/plant and seed weight/plant were detected. Phenotypic variability explained by these QTLs varied from 1 to 15.23%. This study provides framework linkage map of linseed using Indian genotypes, which needs to be enriched further for future application in marker assisted breeding of linseed.     

Mapping quantitative trait loci controlling soybean seed starch content in an interspecific cross of ‘Williams 82’ (Glycine max) and ‘PI 366121’ (Glycine soja)

Seed starch content (SSC) greatly affects the taste, flavour and processing properties of soy foods. The objective in this study was to identify quantitative trait loci (QTL) for SSC in soybean. A total in 169 recombinant inbred lines (RILs) derived from a cross in ‘Williams 82’ and ‘PI 366121’ were grown for three consecutive years. The SSC of the RILs displayed continuous variation with transgressive segregation and hence amenable for QTL mapping. Nine significant QTL exhibiting 5.6–11.3% of the total phenotypic variation (PVE) were identified. The QTL qSTR06_2 showed highest PVE (9.1–11.3%) at LOD values of 4.25–5.39. No stable QTL over 3 years were identified, indicating strong environmental influence on SSC. The QTL qSTR11_1 and qSTR20_1 were found to colocalize with some of the previously reported QTL for sucrose content in soybean, implying the interrelationship between starch and sucrose biosynthesis. As the carbohydrate components may affect key constituents such as oil and protein in soybean seed, findings of the study may be useful in breeding soybeans with improved seed composition.     

甘蓝型油菜胚色素成分的QTL定位

以甘蓝型黄籽油菜GH06和甘蓝型黑籽油菜中油821为亲本杂交,后代通过“一粒传法”连续自交7代构建重组自交系, 2007年分别在重庆市北碚区和万州区两个试验基地种植重组自交系群体, 利用本实验室已构建的遗传连锁图谱和复合区间作图法(CIM), 分析种胚色素的4种主要成分的QTL。结果共检测到31个QTL, 分别位于14个不同的连锁群, 其中5个花色素含量有QTL, 分别位于第1、5、10、16和20连锁群,单个QTL解释表型变异的6.08%~11.67%;10个类黄酮含量有QTL, 分别位于第1、3、6、7、12、20和25连锁群,单个QTL解释表型变异的4.48%~11.10%;8个总酚含量有QTL, 分别位于第1、2、12、16和20连锁群,单个QTL解释表型变异的5.24%~10.37%;8个黑色素含量检测到QTL, 分别位于第5、8、10、12、14和22连锁群,单个QTL解释表型变异的5.44%~11.32%。解释表型变异大于10%的5个QTL, 包括2个类黄酮含量QTL, 花色素含量、总酚含量和黑色素含量QTL各1个,它们分别解释11.10%、10.20%、11.67%、10.37%和11.32%的表型变异。研究结果表明胚色素表现为多基因控制的数量性状, 基因表达受环境影响较大, 胚与种皮色素的QTL吻合度不高, 推测种皮和胚色素合成可能受不同遗传体系控制, 与这些QTL紧密相关的分子标记可以用于胚主要色素的分子标记辅助选择。     

Dissection of genetic architecture for oil content in soybean seed using two backcross populations

Soybean seed oil was valued in foods, animal feed and some industrial applications. Molecular marker‐assisted selection (MAS) for high‐oil‐content cultivars was an important method for soybean breeders. The objective of this study was to identify quantitative trait loci (QTL) and epistatic QTL underlying the seed oil content of soybeans across two backcross (BC) populations (with one common male parent ‘Dongnong47’) and two different environments. Two molecular genetic maps were constructed. They encompassed 1046.8 cM [with an average distance of 6.75 cM in the ‘Dongnong47’  ×  ‘Jiyu89’ (DJ) population] and 846.10 cM [with an average distance of 5.76 cM in the ‘Dongnong47’  ×  ‘Zaoshu18’ (DZ) population]. Nine and seven QTL were identified to be associated with oil content in the DJ and DZ populations, respectively. The phenotypic variation explained by most of the QTL was usually less than 10%. Among the identified QTL, those stable ones across multiple environments and populations often had stronger additive effects. In addition, three stable QTL in the DZ populations were identified in the similar genomic region of the three QTL in the DJ population [qDJE and qDZE‐1 were located near Satt151 of Chromosome 15 (Chr15), qDJA1 and qDZA1 were located near Satt200 of Chr15 (LG A1), and qDJD2‐1 and qDZD2‐1 were located near Sat365 of Chr17]. In conclusion, MAS will be able more effectively to combine beneficial alleles of the different donors to design new genotypes with higher soybean seed oil content using the BC populations.     

Detection of loci controlling seed glucosinolate content and their association with Sclerotinia resistance in Brassica napus

A genetic linkage map of Brassica napus constructed from a cross between a low glucosinolate cultivar ‘H5200’ and a high glucosinolate line ‘NingRS‐1’ was used to identify loci associated with seed glucosinolate content and to understand the association between specific glucosinolate components and Sclerotinia resistance. Seed glucosinolate content was assessed by standard High pressure Liquid Chromatogram (HPLC) protocol. Seven components of seed glucosinolate, including four types of aliphatic glucosinolate, two types of indolyl glucosinolates and one aromatic glucosinolate were detected in the seeds. Three quantitative trait loci (QTLs) were identified for seed total glucosinolate content. From three to 15 loci were found to be responsible for different types of glucosinolates, and by comparing the overlapped intervals, eight genomic regions were defined. One of the nine loci associated with aliphatic glucosinolate content was found to be associated with Sclerotinia resistance on the leaf at the seedling stage, and one locus, responsible for 3‐indolyl‐methyl glucosinolate content, was probably linked with Sclerotinia resistance on the stem of the maturing plant. The association between seed glucosinolate content and Sclerotinia resistance is discussed.