Candidate Quantitative Trait Loci Research Articles

Association mapping of seed quality traits in Brassica napus L. using GWAS and candidate QTL approaches

Single nucleotide polymorphisms (SNPs) have rapidly become the molecular marker of choice in plant and animal association mapping (AM) studies. In this work, a genome-wide association study (GWAS) and candidate quantitative trait loci (cQTL) approaches were used to identify SNP markers associated with seed quality traits, in a Brassica napus L. association panel composed of 89 adapted winter oilseed rape accessions. Six seed quality traits (oil and protein content, linolenic acid, total glucosinolates, hemicellulose and cellulose content) were evaluated in two different locations for two seasons. For GWAS, 4025 SNP markers evenly distributed along the B. napus genome were genotyped using a 6K Illumina array platform. For cQTL, 100 SNP markers previously discovered in genomic regions underlying seed quality QTL were genotyped using a competitive allele-specific PCR (KASPar). Analysis of the population structure revealed the presence of two weakly differentiated subpopulations (F ST = 0.037), with 82 % of the pairwise kinship comparisons ranging from 0 to 0.1. The GWAS approach resulted in the identification of 17 and 5 significant associations for seed glucosinolate content and seed hemicellulose content, respectively. The cQTL approach identified 4 significant associations for seed glucosinolate content and 6 significant associations for seed hemicellulose content. The associated SNPs were consistently identified across environments and were mapped to previously reported QTL. These results illustrate the suitability of AM to identify SNP markers associated with seed quality traits in B. napus.

The first linkage map for a recombinant inbred line population in cotton (Gossypium barbadense) and its use in studies of PEG-induced dehydration tolerance

Egyptian or Pima cotton (Gossypium barbadense L.) is grown for its superior fiber quality in arid and semiarid region where drought is prevalent. However, its genetic and genomic resources are lacking. The objectives of this study were to develop the first recombinant inbred line (RIL) population for linkage mapping and to identify quantitative trait loci (QTLs) for drought tolerance using candidate gene-based single strand conformation polymorphic (SSCP) markers. Out of 152 primer pairs designed from 92 drought-responsive genes, 80 primer pairs yielded 165 polymorphic markers in the Egyptian RIL population of drought tolerant ‘Dandara’ × drought sensitive ‘Giza-70’. Seedlings of the RILs were evaluated for plant height, fresh shoot weight, fresh root weight, chlorophyll content, evapotranspiration, and leaf temperature in two replicated tests in the greenhouse under 10 % polyethylene glycol (PEG) treatment and control (water) conditions. Significant genotypic differences were detected within the RILs, and all the traits were significantly and positively correlated with one another except between fresh root weight and leaf temperature in one test. Based on a linkage map comprised of 247 loci assembled onto 32 linkage groups, 14 QTLs on 11 chromosomes were detected under the control or PEG conditions, each explaining 14–23.5 % of the phenotypic variation. Three chromosomes each carried two QTLs in the same regions, while 7 QTLs were consistent with previous studies. Out of 25 SSCP-SNP gene markers that were significantly correlated with the traits studied, 17 were commonly correlated with more than one trait. This linkage map represents the first for G. barbadense based on a permanent mapping population, and the candidate gene markers and QTLs reported provide useful information for further studies leading to marker-assisted selection and a better understanding of molecular basis in drought tolerance in cotton.

Genetic Mapping in Mice Reveals the Involvement of Pcdh9 in Long-Term Social and Object Recognition and Sensorimotor Development

BackgroundQuantitative genetic analysis of basic mouse behaviors is a powerful tool to identify novel genetic phenotypes contributing to neurobehavioral disorders. Here, we analyzed genetic contributions to single-trial, long-term social and nonsocial recognition and subsequently studied the functional impact of an identified candidate gene on behavioral development. MethodsGenetic mapping of single-trial social recognition was performed in chromosome substitution strains, a sophisticated tool for detecting quantitative trait loci (QTL) of complex traits. Follow-up occurred by generating and testing knockout (KO) mice of a selected QTL candidate gene. Functional characterization of these mice was performed through behavioral and neurological assessments across developmental stages and analyses of gene expression and brain morphology. ResultsChromosome substitution strain 14 mapping studies revealed an overlapping QTL related to long-term social and object recognition harboring Pcdh9, a cell-adhesion gene previously associated with autism spectrum disorder. Specific long-term social and object recognition deficits were confirmed in homozygous (KO) Pcdh9-deficient mice, while heterozygous mice only showed long-term social recognition impairment. The recognition deficits in KO mice were not associated with alterations in perception, multi-trial discrimination learning, sociability, behavioral flexibility, or fear memory. Rather, KO mice showed additional impairments in sensorimotor development reflected by early touch-evoked biting, rotarod performance, and sensory gating deficits. This profile emerged with structural changes in deep layers of sensory cortices, where Pcdh9 is selectively expressed. ConclusionsThis behavior-to-gene study implicates Pcdh9 in cognitive functions required for long-term social and nonsocial recognition. This role is supported by the involvement of Pcdh9 in sensory cortex development and sensorimotor phenotypes.

Large effect QTL explain natural phenotypic variation for the developmental timing of vegetative phase change in maize (Zea mays L.).

Natural variation for the timing of vegetative phase change in maize is controlled by several large effect loci, one corresponding to Glossy15 , a gene known for regulating juvenile tissue traits. Vegetative phase change is an intrinsic component of developmental programs in plants. Juvenile and adult vegetative tissues in grasses differ dramatically in their anatomical and biochemical composition affecting the utility of specific genotypes as animal feed and biofuel feedstock. The molecular network controlling the process of developmental transition is incompletely characterized. In this study, we used scoring for juvenile and adult epicuticular wax as an entry point to discover quantitative trait loci (QTL) controlling phenotypic variation for the developmental timing of juvenile to adult transition in maize. We scored the last leaf with juvenile wax on 25 recombinant inbred line families of the B73 reference Nested Association Mapping (NAM) population and the intermated B73×Mo17 (IBM) population across multiple seasons. A total of 13 unique QTL were identified through genome-wide association analysis across the NAM populations, three of which have large effects. A QTL located on chromosome nine had the most significant SNPs within Glossy15, a gene controlling expression of juvenile leaf traits. The second large effect QTL is located on chromosome two. The most significant SNP in this QTL is located adjacent to a homolog of the Arabidopsis transcription factor, enhanced downy mildew-2, which has been shown to promote the transition from juvenile to adult vegetative phase. Overall, these results show that several major QTL and potential candidate genes underlie the extensive natural variation for this developmental trait.

Mapping quantitative trait loci for nitrogen uptake and utilization efficiency in rice (Oryza sativa L.) at different nitrogen fertilizer levels.

Genetic improvement is the fundamental basis for improving nitrogen-use efficiency. A better understanding of genetic factors controlling nitrogen uptake and utilization is required for crop genetic improvement. In this study, we identified the quantitative trait loci (QTLs) associated with traits of nitrogen uptake and utilization by using the single-sequence repeat marker method and a recombinant inbred line (RIL) population derived from a super hybrid Xieyou9308. All the traits investigated were inherited quantitatively by continuous variation and showed normal distribution in phenotype with transgressive segregation in the RIL population. Most of the traits were significantly correlated with each other except for nitrogen absorption ability (NAA) with nitrogen harvest index (NHI) and NHI with agricultural nitrogen-absorption efficiency (ANAE). At logarithmic odds value of 2.3, total 13 candidate QTLs, including 4 for NAA, 2 for NHI, 2 for physiological nitrogen-use efficiency, 1 for agricultural nitrogen-use efficiency (ANUE), and 4 for ANAE, were detected and mapped on chromosomes 2, 3, 4, 5, 8, 9, 10, and 12. Significant pleiotropic effect or neighboring expression of QTLs was observed among traits. At position 64.8 cM on chromosome 4 near the marker RM5757, there was a QTL cluster of NAA, ANUE, and ANAE, and at chromosome 5 near the marker RM5968, there was a QTL cluster of NAA and ANUE. The QTL clusters might provide partial explanation and genetic mechanism for the observed correlations between nitrogen uptake and utilization efficiency traits and might form a basis for future breeding programs.

Linkage disequilibrium with linkage analysis of multiline crosses reveals different multiallelic QTL for hybrid performance in the flint and dent heterotic groups of maize.

Multiparental designs combined with dense genotyping of parents have been proposed as a way to increase the diversity and resolution of quantitative trait loci (QTL) mapping studies, using methods combining linkage disequilibrium information with linkage analysis (LDLA). Two new nested association mapping designs adapted to European conditions were derived from the complementary dent and flint heterotic groups of maize (Zea mays L.). Ten biparental dent families (N = 841) and 11 biparental flint families (N = 811) were genotyped with 56,110 single nucleotide polymorphism markers and evaluated as test crosses with the central line of the reciprocal design for biomass yield, plant height, and precocity. Alleles at candidate QTL were defined as (i) parental alleles, (ii) haplotypic identity by descent, and (iii) single-marker groupings. Between five and 16 QTL were detected depending on the model, trait, and genetic group considered. In the flint design, a major QTL (R(2) = 27%) with pleiotropic effects was detected on chromosome 10, whereas other QTL displayed milder effects (R(2) < 10%). On average, the LDLA models detected more QTL but generally explained lower percentages of variance, consistent with the fact that most QTL display complex allelic series. Only 15% of the QTL were common to the two designs. A joint analysis of the two designs detected between 15 and 21 QTL for the five traits. Of these, between 27 for silking date and 41% for tasseling date were significant in both groups. Favorable allelic effects detected in both groups open perspectives for improving biomass production.

Identification of a QTL in Mus musculus for alcohol preference, withdrawal, and Ap3m2 expression using integrative functional genomics and precision genetics.

Extensive genetic and genomic studies of the relationship between alcohol drinking preference and withdrawal severity have been performed using animal models. Data from multiple such publications and public data resources have been incorporated in the GeneWeaver database with >60,000 gene sets including 285 alcohol withdrawal and preference-related gene sets. Among these are evidence for positional candidates regulating these behaviors in overlapping quantitative trait loci (QTL) mapped in distinct mouse populations. Combinatorial integration of functional genomics experimental results revealed a single QTL positional candidate gene in one of the loci common to both preference and withdrawal. Functional validation studies in Ap3m2 knockout mice confirmed these relationships. Genetic validation involves confirming the existence of segregating polymorphisms that could account for the phenotypic effect. By exploiting recent advances in mouse genotyping, sequence, epigenetics, and phylogeny resources, we confirmed that Ap3m2 resides in an appropriately segregating genomic region. We have demonstrated genetic and alcohol-induced regulation of Ap3m2 expression. Although sequence analysis revealed no polymorphisms in the Ap3m2-coding region that could account for all phenotypic differences, there are several upstream SNPs that could. We have identified one of these to be an H3K4me3 site that exhibits strain differences in methylation. Thus, by making cross-species functional genomics readily computable we identified a common QTL candidate for two related bio-behavioral processes via functional evidence and demonstrate sufficiency of the genetic locus as a source of variation underlying two traits.

SIRPB1 copy-number polymorphism as candidate quantitative trait locus for impulsive-disinhibited personality.

Impulsive-disinhibited personality (IDP) is a behavioral trait mainly characterized by seeking immediate gratification at the expense of more enduring or long-term gains. This trait has a major role in the development of several disinhibitory behaviors and syndromes, including psychopathy, attention-deficit and hyperactivity disorder, cluster-B personality disorders, criminality and alcoholism. Available data consistently support a strong heritable component, accounting for 30-60% of the observed variance in personality traits. A genome-wide analysis of copy-number variants was designed to identify novel genetic pathways associated with the IDP trait, using a series of 261 male participants with maximized opposite IDP scores. Quantitative trait locus analysis of candidate copy-number variants (CNVs) was conducted across the entire IDP continuum. Functional effects of associated variants were evaluated in zebrafish embryos. A common CNV mapping to the immune-related gene SIRPB1 was significantly associated with IDP scores in a dose-dependent manner (β=-0.172, P<0.017). Expression quantitative trait locus analysis of the critical region revealed higher SIRPB1 mRNA levels associated with the haplotype containing the deleted allele (P<0.0007). Epigenetic marks highlighted the presence of two potential insulators within the deleted region, confirmed by functional assays in zebrafish embryos, which suggests that SIRPB1 expression rates are affected by the presence/absence of the insulator regions. Upregulation of SIRPB1 has been described in prefrontal cortex of patients with schizophrenia, providing a link between SIRPB1 and diseases involving disinhibition and failure to control impulsivity. We propose SIRPB1 as a novel candidate gene to account for phenotypic differences observed in the IDP trait.

Association of the Fusarium Wilt Race 3 Resistance Gene, I-3, on Chromosome 7 with Increased Susceptibility to Bacterial Spot Race T4 in Tomato

Bacterial spot of tomato (Solanum lycopersicum), caused by several Xanthomonas species, is one of the most important diseases of the crop in humid production regions of the world. Conventional breeding approaches for resistance to bacterial spot previously identified race-specific resistances, but current efforts also seek to use quantitative trait loci (QTLs) effecting broad-spectrum resistance. Resistance QTLs and candidate QTLs have been reported on several chromosomes, including a major QTL on chromosome 11. Fusarium wilt (Fusarium oxysporum f. sp. lycopersici) race 3 resistance gene, I-3, is associated with smaller fruit size and has been implicated in other associations with negative characteristics. We evaluated four F2 populations involving the bacterial spot-tolerant breeding lines Fla. 8517, Fla. 8233, and Fla. 8326 across two field seasons to validate and quantify previously identified loci and to test for an effect of I-3 on bacterial spot sensitivity. The chromosome 11 QTL and the I-3 locus were each consistently positively and negatively associated with resistance, respectively, and together explained from 44% to 47% of the variation in each population. The chromosome 11 QTL displayed a dominant to incompletely dominant effect, reducing infection by 14% to 25%. This QTL is distinct from the X. perforans race T3 hypersensitivity loci, Rx-4 and Xv3. The I-3 locus contributed to as much as a 20% increase in infection in I-3/I-3 plants vs. i-3/i-3 plants, and heterozygosity for I-3 generally resulted in an intermediate susceptible response. Significant effects for QTLs on chromosomes 3, 5, and 12 were also observed, but these effects were not consistent in all populations or seasons in which they were segregating. Implications of these findings toward breeding strategies are discussed.

Genomic structure of nucleotide diversity among Lyon rat models of metabolic syndrome

BackgroundThe metabolic syndrome (MetS), a complex disorder involving hypertension, obesity, dyslipidemia and insulin resistance, is a major risk factor for heart disease, stroke, and diabetes. The Lyon Hypertensive (LH), Lyon Normotensive (LN) and Lyon Low-pressure (LL) rats are inbred strains simultaneously derived from a common outbred Sprague Dawley colony by selection for high, normal, and low blood pressure, respectively. Further studies found that LH is a MetS susceptible strain, while LN is resistant and LL has an intermediate phenotype. Whole genome sequencing determined that, while the strains are phenotypically divergent, they are nearly 98% similar at the nucleotide level. Using the sequence of the three strains, we applied an approach that harnesses the distribution of Observed Strain Differences (OSD), or nucleotide diversity, to distinguish genomic regions of identity-by-descent (IBD) from those with divergent ancestry between the three strains. This information was then used to fine-map QTL identified in a cross between LH and LN rats in order to identify candidate genes causing the phenotypes.ResultsWe identified haplotypes that, in total, contain at least 95% of the identifiable polymorphisms between the Lyon strains that are likely of differing ancestral origin. By intersecting the identified haplotype blocks with Quantitative Trait Loci (QTL) previously identified in a cross between LH and LN strains, the candidate QTL regions have been narrowed by 78%. Because the genome sequence has been determined, we were further able to identify putative functional variants in genes that are candidates for causing the QTL.ConclusionsWhole genome sequence analysis between the LH, LN, and LL strains identified the haplotype structure of these three strains and identified candidate genes with sequence variants predicted to affect gene function. This approach, merged with additional integrative genetics approaches, will likely lead to novel mechanisms underlying complex disease and provide new drug targets and therapies.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-197) contains supplementary material, which is available to authorized users.

Association mapping of seed quality traits using the Canadian flax (Linum usitatissimum L.) core collection.

Key message The identification of stable QTL for seed quality traits by association mapping of a diverse panel of linseed accessions establishes the foundation for assisted breeding and future fine mapping in linseed. Linseed oil is valued for its food and non-food applications. Modifying its oil content and fatty acid (FA) profiles to meet market needs in a timely manner requires clear understanding of their quantitative trait loci (QTL) architectures, which have received little attention to date. Association mapping is an efficient approach to identify QTL in germplasm collections. In this study, we explored the quantitative nature of seed quality traits including oil content (OIL), palmitic acid, stearic acid, oleic acid, linoleic acid (LIO) linolenic acid (LIN) and iodine value in a flax core collection of 390 accessions assayed with 460 microsatellite markers. The core collection was grown in a modified augmented design at two locations over 3 years and phenotypic data for all seven traits were obtained from all six environments. Significant phenotypic diversity and moderate to high heritability for each trait (0.73–0.99) were observed. Most of the candidate QTL were stable as revealed by multivariate analyses. Nine candidate QTL were identified, varying from one for OIL to three for LIO and LIN. Candidate QTL for LIO and LIN co-localized with QTL previously identified in bi-parental populations and some mapped nearby genes known to be involved in the FA biosynthesis pathway. Fifty-eight percent of the QTL alleles were absent (private) in the Canadian cultivars suggesting that the core collection possesses QTL alleles potentially useful to improve seed quality traits. The candidate QTL identified herein will establish the foundation for future marker-assisted breeding in linseed.Electronic supplementary materialThe online version of this article (doi:10.1007/s00122-014-2264-4) contains supplementary material, which is available to authorized users.

Dynamic QTL analysis and candidate gene mapping for waterlogging tolerance at maize seedling stage.

Soil waterlogging is one of the major abiotic stresses adversely affecting maize growth and yield. To identify dynamic expression of genes or quantitative trait loci (QTL), QTL associated with plant height, root length, root dry weight, shoot dry weight and total dry weight were identified via conditional analysis in a mixed linear model and inclusive composite interval mapping method at three respective periods under waterlogging and control conditions. A total of 13, 19 and 23 QTL were detected at stages 3D|0D (the period during 0–3 d of waterlogging), 6D|3D and 9D|6D, respectively. The effects of each QTL were moderate and distributed over nine chromosomes, singly explaining 4.14–18.88% of the phenotypic variation. Six QTL (ph6-1, rl1-2, sdw4-1, sdw7-1, tdw4-1 and tdw7-1) were identified at two consistent stages of seedling development, which could reflect a continuous expression of genes; the remaining QTL were detected at only one stage. Thus, expression of most QTL was influenced by the developmental status. In order to provide additional evidence regarding the role of corresponding genes in waterlogging tolerance, mapping of Expressed Sequence Tags markers and microRNAs were conducted. Seven candidate genes were observed to co-localize with the identified QTL on chromosomes 1, 4, 6, 7 and 9, and may be important candidate genes for waterlogging tolerance. These results are a good starting point for understanding the genetic basis for selectively expressing of QTL in different stress periods and the common genetic control mechanism of the co-localized traits.

High-Resolution Linkage and Quantitative Trait Locus Mapping Aided by Genome Survey Sequencing: Building Up An Integrative Genomic Framework for a Bivalve Mollusc

Genetic linkage maps are indispensable tools in genetic and genomic studies. Recent development of genotyping-by-sequencing (GBS) methods holds great promise for constructing high-resolution linkage maps in organisms lacking extensive genomic resources. In the present study, linkage mapping was conducted for a bivalve mollusc (Chlamys farreri) using a newly developed GBS method—2b-restriction site-associated DNA (2b-RAD). Genome survey sequencing was performed to generate a preliminary reference genome that was utilized to facilitate linkage and quantitative trait locus (QTL) mapping in C. farreri. A high-resolution linkage map was constructed with a marker density (3806) that has, to our knowledge, never been achieved in any other molluscs. The linkage map covered nearly the whole genome (99.5%) with a resolution of 0.41 cM. QTL mapping and association analysis congruously revealed two growth-related QTLs and one potential sex-determination region. An important candidate QTL gene named PROP1, which functions in the regulation of growth hormone production in vertebrates, was identified from the growth-related QTL region detected on the linkage group LG3. We demonstrate that this linkage map can serve as an important platform for improving genome assembly and unifying multiple genomic resources. Our study, therefore, exemplifies how to build up an integrative genomic framework in a non-model organism.

Soybean Seed Amino Acid Content QTL Detected Using the Uni - versal Soy Linkage Panel 1.0 with 1,536 SNPs

Soybean [Glycine max (L.) Merr.] is the primary source of meal used in animal feed in the U.S. However, few studies have been conducted to evaluate genomic regions controlling amino acid composition is soybean. Designing soybean seed compositions that will benefit animal production is essential. The objective of this study was to identify genomic regions controlling essential and non-essential amino acid composition in soybean seed proteins. To achieve this objective, 282 F5:9 recombinant inbred lines (RILs) developed from a cross of Essex × Williams 82 were used. Ground soybean seed samples were analyzed for amino acids and statistically significant differences (p &lt; 0.05) were found among genotypes in the population for all amino acid concentrations. The Universal Soy Linkage Panel (USLP) 1.0 of 1,536 single nucleotide polymorphism (SNP) DNA markers were used to genotype the 282 RILs and identify 480 useful genetic markers. The software R/qtl was used to identify candidate quantitative trait loci (QTL), which were validated using R/MQM. A total of ten QTL were detected on chromosomes 5, 7, 9, 10, 13 and 20 that explained 5 to 14% of the total phenotypic variation for a particular amino acid. Using SNPs from the USLP 1.0 to detect QTL for amino acids in soybean provides additional information to select genotypes with enhanced amino acid profiles that will benefit animal production.

Characterization of growth hormone secretagogue receptor 1 (GHS-R1) genes and weight gain associated SNP loci in Cyprinus carpio var. jian

Growth hormone secretagogue receptors(GHS-Rs) are endogenous receptors for growth hormone se-cretion(ghrelin) that belong to the G-protein-coupled receptor family.GHS-Rs play a role in regulating animal growth and energy homeostasis.GHS-R is a candidate quantitative trait loci related to obesity and growth in mammals.We used RT-PCR and PCR to isolate two jlGHS-R1s,jlGHS-R1a and 1b.The open reading frames of jlGHS-R1s encode 360 amino acids that share 96% identity.In addition,there are two jlGHS-R1s transcription variants,an alternatively spliced 191 bp fragment from 490 nt to 680 nt in ORF with GT-AG at both ends.These transcripts led to a premature termination of translation,encoding 184 aa,and only contained three and a half transmembrane regions,which differs from the reported intron retaining variants among tilapia.The ORF of jlGHS-R1s was separated by one intron,locating between the first and second base of A260 coden.The introns of 1a and 1b were 676 bp and 885 bp in length,respectively.We found 32 SNPs on two jlGHS-R1s in the Cyprinus carpio var.jian population using alignment sequences from different individuals.We then genotyped 9 SNPs using PCR-RFLP.We encountered each genotype in 322 individuals,but there was an obvious bias in the distribution.The 1a-I_C386T and 1b-E1_G159T sites were significantly associated with juvenile and adult fish weight gain(P0.01 and P0.05,respectively).Individuals with CC and GG genotypes grew faster than other individuals.In addition,a further five sites were correlated with weight gain at certain stages or sex.To determine the applicabil-ity of the markers obtained in this experiment,we tested 610 individuals from seven additional families using four markers.The C386T and G159T sites remained significantly correlated with weight gain.Therefore,these two sites can be used as references for the molecular breeding of Cyprinus carpio var.jian.

Efficacy of pyramiding elite alleles for dynamic development of plant height in common wheat

Plant height is an important botanical feature closely related to yield. Two populations consisting of 118 and 262 accessions respectively were used to identify elite alleles for plant height and to validate their allelic effects. Plant height was measured from the early booting to the flowering stages. Simple sequence repeat markers for candidate quantitative trait locus (QTL) regions with large effects identified in a doubled haploid (DH) population (Hanxuan 10 × Lumai 14) were selected for further verification by association analysis. Nine loci significantly (P < 0.001) associated with plant height were detected 13 times in the population with 118 accessions. Three loci (Xgwm11-1B, Xwmc349-4B and Xcfd23-4D) were identified in three, two and two periods of plant height growth, respectively. Markers Xbarc168-2D, Xgwm249-2D, Xwmc349-4B, Xcfd23-4D and Xgwm410-5A located at or near additive QTL regions in the DH population proved to coincide with known Rht loci. The results showed a consistency between linkage analysis and association mapping, and also confirmed the value of fine mapping of QTL through combined linkage and association analyses. For final plant height, the alleles Xgwm11-1B 208, Xwmc349-4B 103 and Xcfd23-4D 202 exhibited negative effects, i.e. reducing plant height; Xwmc349-4B 101 and Xcfd23-4D 205 showed significant positive effects. A second larger population (262 accessions) was used to validate the effects of these large-effect alleles and the efficacy of pyramiding in eight environments (year × site × water regime combinations). Strong correlations between final plant height and numbers of large-effect alleles indicated that the alleles contributed additively to plant height. The additive effects showed that pyramiding elite alleles for target traits has significant potential for wheat breeding.Electronic supplementary materialThe online version of this article (doi:10.1007/s11032-013-9873-5) contains supplementary material, which is available to authorized users.

Quantitative trait locus linkage analysis in a large Amish pedigree identifies novel candidate loci for erythrocyte traits

We characterized a large Amish pedigree and, in 384 pedigree members, analyzed the genetic variance components with covariate screen as well as genome-wide quantitative trait locus (QTL) linkage analysis of red blood cell count (RBC), hemoglobin (HB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), platelet count (PLT), and white blood cell count (WBC) using SOLAR. Age and gender were found to be significant covariates in many CBC traits. We obtained significant heritability estimates for RBC, MCV, MCH, MCHC, RDW, PLT, and WBC. We report four candidate loci with Logarithm of the odds (LOD) scores above 2.0: 6q25 (MCH), 9q33 (WBC), 10p12 (RDW), and 20q13 (MCV). We also report eleven candidate loci with LOD scores between 1.5 and <2.0. Bivariate linkage analysis of MCV and MCH on chromosome 20 resulted in a higher maximum LOD score of 3.14. Linkage signals on chromosomes 4q28, 6p22, 6q25, and 20q13 are concomitant with previously reported QTL. All other linkage signals reported herein represent novel evidence of candidate QTL. Interestingly rs1800562, the most common causal variant of hereditary hemochromatosis in HFE (6p22) was associated with MCH and MCHC in this family. Linkage studies like the one presented here will allow investigators to focus the search for rare variants amidst the noise encountered in the large amounts of data generated by whole-genome sequencing.

Recalculation of 23 mouse HDL QTL datasets improves accuracy and allows for better candidate gene analysis

In the past 15 years, the quantitative trait locus (QTL) mapping approach has been applied to crosses between different inbred mouse strains to identify genetic loci associated with plasma HDL cholesterol levels. Although successful, a disadvantage of this method is low mapping resolution, as often several hundred candidate genes fall within the confidence interval for each locus. Methods have been developed to narrow these loci by combining the data from the different crosses, but they rely on the accurate mapping of the QTL and the treatment of the data in a consistent manner. We collected 23 raw datasets used for the mapping of previously published HDL QTL and reanalyzed the data from each cross using a consistent method and the latest mouse genetic map. By utilizing this approach, we identified novel QTL and QTL that were mapped to the wrong part of chromosomes. Our new HDL QTL map allows for reliable combining of QTL data and candidate gene analysis, which we demonstrate by identifying Grin3a and Etv6, as candidate genes for QTL on chromosomes 4 and 6, respectively. In addition, we were able to narrow a QTL on Chr 19 to five candidates.

Detection of QTLs to reduce cadmium content in rice grains using LAC23/Koshihikari chromosome segment substitution lines

To advance the identification of quantitative trait loci (QTLs) to reduce Cd content in rice (Oryza sativa L.) grains and breed low-Cd cultivars, we developed a novel population consisting of 46 chromosome segment substitution lines (CSSLs) in which donor segments of LAC23, a cultivar reported to have a low grain Cd content, were substituted into the Koshihikari genetic background. The parental cultivars and 32 CSSLs (the minimum set required for whole-genome coverage) were grown in two fields with different natural levels of soil Cd. QTL mapping by single-marker analysis using ANOVA indicated that eight chromosomal regions were associated with grain Cd content and detected a major QTL (qlGCd3) with a high F-test value in both fields (F = 9.19 and 5.60) on the long arm of chromosome 3. The LAC23 allele at qlGCd3 was associated with reduced grain Cd levels and appeared to reduce Cd transport from the shoots to the grains. Fine substitution mapping delimited qlGCd3 to a 3.5-Mbp region. Our results suggest that the low-Cd trait of LAC23 is controlled by multiple QTLs, and qlGCd3 is a promising candidate QTL to reduce the Cd level of rice grain.

Genotype – environment correlations in corals from the Great Barrier Reef

BackgroundKnowledge of genetic markers that are correlated to stress tolerance may improve spatial mapping of reef vulnerability and can inform restoration efforts, including the choice of genotypes for breeding and reseeding. In this manuscript we present two methods for screening transcriptome data for candidate genetic markers in two reef building corals, Acropora millepora and Pocillopora damicornis (types α and β). In A. millepora, Single Nucleotide Polymorphisms (SNPs) were pre-selected by targeting genes believed to be involved in the coral thermal stress responses. In P. damicornis (type α and β), SNPs showing varying allele frequencies between two populations from distinct environments were pre-selected. Allele frequencies at nine, five and eight of the pre-selected SNP loci were correlated against gradients of water clarity and temperature in a large number of populations along the Great Barrier Reef.ResultsA significant correlation between environmental category and SNP allele frequency was detected in up to 55% of the tested loci, which is an exceptional success rate for these types of tests. In P. damicornis, SNP allele frequencies of β-hexosaminidase and Elongation factor 1-α were significantly correlated to temperature in type α and to temperature and/or water clarity respectively in type β. Type α also showed a correlation between water clarity and SNP allele frequency in a gene of unknown function. In A. millepora, allele frequencies at five (β-gamma crystallin, Galaxin, Ubiquitin, Ligand of Numb X2 and Thioredoxin) SNP loci showed significant correlations.ConclusionsAfter validation of these candidate loci through laboratory or field assessment of relative stress tolerance of colonies harbouring different alleles, it is anticipated that a proportion of these markers may represent the first coral candidate Quantitative Trait Loci for environmental stress tolerance and provide an important genetic tool that can be incorporated into spatial management decisions and restoration efforts of coral reefs. One pertinent example would be to combine spatial data of tolerant populations with genetic connectivity and thus identify high priority conservation reefs and implement targeted coral husbandry and active restoration efforts that use locally- and stress-adapted genotypes.