Genome-wide Association Study Mapping Research Articles

Genetic associations of transgenerational stress memory in wheat under drought stress

Transgenerational stress memory (TSM) in plants is a fascinating area of research, particularly when it comes to understanding how plants respond to drought stress. Therefore, our study explored the genetic architecture/causative alleles controlling transgenerational drought stress memoryin a diverse collection of 111 wheat accessions that enhanced seed germination parameters and antioxidant components in response to drought stress tolerance using a Genome-Wide Association Study (GWAS). This experiment was performed in two distinct stages. In the first stage, all wheat accessions were exposed to control and drought conditions following the primed acclimation technique. Two different groups of genotypes were recovered at this stage: the seeds of stressed plants (SP) and those of non-stressed plants (NP) and evaluated under drought treatment. Our study revealed a highly significant increase in root and shoot lengths by 42 % and 56 % for the seeds of stressed plants Similarly, a highly significant increase in superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase was detected for stressed plants by 55 %, 43 %, 44 %, and 63 % when compared to those of non-stressed wheatplants. Using GWAS mapping, a significant marker (Kukri_c53629_239) associated with APX_SP, DW_SP, and SOD_SP on chromosome 2 H was located inside the gene TraesCS2B02G192700 candidate is annotated as protein kinase activity that triggering various protective mechanisms, such as antioxidative enzymes under drought stress. Altogether, TSM is a cornerstone in the genetic research of drought stress tolerance, offering invaluable insights that can drive the development of drought-resilient crop varieties.

Fast integration and accumulation of beneficial breeding alleles through an AB–NAMIC strategy in wheat

Wheat (Triticum aestivum) is among the most important staple crops for safeguarding the food security of the growing world population. To bridge the gap between genebank diversity and breeding programs, we developed an advanced backcross–nested association mapping plus inter-crossed population (AB–NAMIC) by crossing three popular wheat cultivars as recurrent founders to 20 germplasm lines from a mini core collection. Selective backcrossing combined with selection against undesirable traits and extensive crossing within and between sub-populations created new opportunities to detect unknown genes and increase the frequency of beneficial alleles in the AB–NAMIC population. We performed phenotyping of 590 AB–NAMIC lines and a natural panel of 476 cultivars for six consecutive growing seasons and genotyped these 1066 lines with a 660K SNP array. Genome-wide association studies of both panels for plant development and yield traits demonstrated improved power to detect rare alleles and loci with medium genetic effects in AB–NAMIC. Notably, genome-wide association studies in AB–NAMIC detected the candidate gene TaSWEET6-7B (TraesCS7B03G1216700), which has high homology to the rice SWEET6b gene and exerts strong effects on adaptation and yield traits. The commercial release of two derived AB–NAMIC lines attests to its direct applicability in wheat improvement. Valuable information on genome-wide association study mapping, candidate genes, and their haplotypes for breeding traits are available through WheatGAB. Our research provides an excellent framework for fast-tracking exploration and accumulation of beneficial alleles stored in genebanks.

Genome-wide association mapping for component traits of drought and heat tolerance in wheat.

Identification of marker trait association is a prerequisite for marker-assisted breeding. To find markers linked with traits under heat and drought stress in bread wheat (Triticum aestivum L.), we performed a genome-wide association study (GWAS). GWAS mapping panel used in this study consists of advanced breeding lines from the IARI stress breeding programme produced by pairwise and complex crosses. Phenotyping was done at multi locations namely New Delhi, Karnal, Indore, Jharkhand and Pune with augmented-RCBD design under different moisture and heat stress regimes, namely timely sown irrigated (IR), timely sown restricted irrigated (RI) and late sown (LS) conditions. Yield and its component traits, viz., Days to Heading (DH), Days to Maturity (DM), Normalized Difference Vegetation Index (NDVI), Chlorophyll Content (SPAD), Canopy temperature (CT), Plant Height (PH), Thousand grain weight (TGW), Grain weight per spike (GWPS), Plot Yield (PLTY) and Biomass (BMS) were phenotyped. Analysis of variance and descriptive statistics revealed significant differences among the studied traits. Genotyping was done using the 35k SNP Wheat Breeder's Genotyping Array. Population structure and diversity analysis using filtered 10,546 markers revealed two subpopulations with sufficient diversity. A large whole genome LD block size of 7.15 MB was obtained at half LD decay value. Genome-wide association search identified 57 unique markers associated with various traits across the locations. Twenty-three markers were identified to be stable, among them nine pleiotropic markers were also identified. In silico search of the identified markers against the IWGSC ref genome revealed the presence of a majority of the SNPs at or near the gene coding region. These SNPs can be used for marker-assisted transfer of genes/QTLs after validation to develop climate-resilient cultivars.

Genome-wide association study identifies loci, beneficial alleles, and candidate genes for cadmium tolerance in castor (Ricinus communis L.)

Cadmium (Cd) is a phytotoxic element that may impede oilseed production. Breeding of Cd-tolerant cultivars is an imperative that can subsequently be used for phytoremediation of Cd-contaminated soils. Hence, to identify marker-trait associations and mine for candidate genes that confer Cd tolerance in castor, genome-wide association study (GWAS) for seven Cdtolerance-related traits was performed using a naturalized population of 175 castor accessions genotyped by 143 polymorphic simple sequence repeat markers. The results showed that there was a wide variation of Cd among the seven traits in the 175 castor accessions. A total of 27 significant (-log10P > 1.5) marker loci were detected to be associated with all the seven traits for Cd tolerance, and the phenotypic variance explained by these loci varied from 5.63∼20.47 %. Mining of beneficial alleles revealed 29 and 181 alleles that contributed prominently to Cd tolerance and sensitivity, respectively, which would be useful for breeding Cd-tolerant cultivars. Eight novel putative candidate genes involved in stress responses and developmental processes were detected within the major loci. Gene expression analyses using the quantitative real-time polymerase chain reaction method on six selected candidate genes showed that all the genes were highly expressed in the tolerant and sensitive accessions under Cd stress, signifying their potential role in enhancing stress tolerance or responses. This study represented the first attempt to GWAS mapping and identification of novel genes in castor under Cd stress. Our results may facilitate the study of the molecular and genetic basis of Cd tolerance and the cloning of candidate Cd-tolerance genes and cultivars, which could be useful for remediation of Cd-contaminated environments.

Meta-analysis for milk fat and protein percentage using imputed sequence variant genotypes in 94,321 cattle from eight cattle breeds

BackgroundSequence-based genome-wide association studies (GWAS) provide high statistical power to identify candidate causal mutations when a large number of individuals with both sequence variant genotypes and phenotypes is available. A meta-analysis combines summary statistics from multiple GWAS and increases the power to detect trait-associated variants without requiring access to data at the individual level of the GWAS mapping cohorts. Because linkage disequilibrium between adjacent markers is conserved only over short distances across breeds, a multi-breed meta-analysis can improve mapping precision.ResultsTo maximise the power to identify quantitative trait loci (QTL), we combined the results of nine within-population GWAS that used imputed sequence variant genotypes of 94,321 cattle from eight breeds, to perform a large-scale meta-analysis for fat and protein percentage in cattle. The meta-analysis detected (p ≤ 10−8) 138 QTL for fat percentage and 176 QTL for protein percentage. This was more than the number of QTL detected in all within-population GWAS together (124 QTL for fat percentage and 104 QTL for protein percentage). Among all the lead variants, 100 QTL for fat percentage and 114 QTL for protein percentage had the same direction of effect in all within-population GWAS. This indicates either persistence of the linkage phase between the causal variant and the lead variant across breeds or that some of the lead variants might indeed be causal or tightly linked with causal variants. The percentage of intergenic variants was substantially lower for significant variants than for non-significant variants, and significant variants had mostly moderate to high minor allele frequencies. Significant variants were also clustered in genes that are known to be relevant for fat and protein percentages in milk.ConclusionsOur study identified a large number of QTL associated with fat and protein percentage in dairy cattle. We demonstrated that large-scale multi-breed meta-analysis reveals more QTL at the nucleotide resolution than within-population GWAS. Significant variants were more often located in genic regions than non-significant variants and a large part of them was located in potentially regulatory regions.

Genome-wide association studies of molybdenum and selenium concentrations in C. arietinum and C. reticulatum seeds

Chickpea is the second most important and ancient pulse crop with its use in human diet for approximately 7500 years as one of the Neolithic founder crops. Previous studies on chickpea have predominantly focused on agronomic traits, with only limited research being dedicated to developing micronutrient-rich crops over the last half century. The objectives of this study were to identify genetic variation and population structure of Cicer reticulatum (C. reticulatum) and Cicer arietinum (C. arietinum) species and reveal marker-trait associations of seed molybdenum (Mo) and selenium (Se) concentrations in seeds by genome-wide association studies (GWAS). In this study, a population of 180 individuals including 107 wild (C. reticulatum) and 73 cultivated (C. arietinum) Cicer species was used in two different locations for 2 years, and 121,840 high-quality single nucleotide polymorphism (SNP) were identified across eight chromosomes through genotyping by sequencing (GBS) analysis. GWAS was performed for 180 individuals and alternatively two subpopulations separately, and 16 SNP markers were found significantly associated with seed Mo and Se concentrations, consistently among the four environments. The results demonstrated the high potential of GWAS mapping in revealing markers associated with Mo and Se micronutrients for wild (C. reticulatum) and cultivated (C. arietinum) species.

Assessment of Five Chilling Tolerance Traits and GWAS Mapping in Rice Using the USDA Mini-Core Collection.

Rice (Oryza sativa L.) is often exposed to cool temperatures during spring planting in temperate climates. A better understanding of genetic pathways regulating chilling tolerance will enable breeders to develop varieties with improved tolerance during germination and young seedling stages. To dissect chilling tolerance, five assays were developed; one assay for the germination stage, one assay for the germination and seedling stage, and three for the seedling stage. Based on these assays, five chilling tolerance indices were calculated and assessed using 202 O. sativa accessions from the Rice Mini-Core (RMC) collection. Significant differences between RMC accessions made the five indices suitable for genome-wide association study (GWAS) based quantitative trait loci (QTL) mapping. For young seedling stage indices, japonica and indica subspecies clustered into chilling tolerant and chilling sensitive accessions, respectively, while both subspecies had similar low temperature germinability distributions. Indica subspecies were shown to have chilling acclimation potential. GWAS mapping uncovered 48 QTL at 39 chromosome regions distributed across all 12 rice chromosomes. Interestingly, there was no overlap between the germination and seedling stage QTL. Also, 18 QTL and 32 QTL were in regions discovered in previously reported bi-parental and GWAS based QTL mapping studies, respectively. Two novel low temperature seedling survivability (LTSS)–QTL, qLTSS3-4 and qLTSS4-1, were not in a previously reported QTL region. QTL with strong effect alleles identified in this study will be useful for marker assisted breeding efforts to improve chilling tolerance in rice cultivars and enhance gene discovery for chilling tolerance.

Genome-wide association study on stem rust resistance in Kazakh spring barley lines.

BackgroundStem rust (SR) is one of the most economically devastating barley diseases in Kazakhstan, and in some years it causes up to 50 % of yield losses. Routine conventional breeding for resistance to stem rust is almost always in progress in all Kazakhstan breeding stations. However, molecular marker based approach towards new SR resistance genes identification and relevant marker-assisted selection had never been employed in Kazakhstan yet. In this study, as a preliminary step the GWAS (genome-wide association study) mapping was applied in attempt to identify reliable SNP markers and quantitative trait loci (QTL) associated with resistance to SR.ResultsBarley collection of 92 commercial cultivars and promising lines was genotyped using a high-throughput single nucleotide polymorphism (9,000 SNP) Illumina iSelect array. 6,970 SNPs out of 9,000 total were polymorphic and scorable. 5,050 SNPs out of 6,970 passed filtering threshold and were used for association mapping (AM). All 92 accessions were phenotyped for resistance to SR by observing adult plants in artificially infected plots at the Research Institute for Biological Safety Problems in Dzhambul region of Kazakhstan. GLM analysis allowed the identification of 15 SNPs associated with the resistance at the heading time (HA) phase, and 2 SNPs at the seed’s milky-waxy maturity (SM) phase. However, after application of 5 % Bonferroni multiple test correction, only 2 SNPs at the HA stage on the same position of chromosome 6H can be claimed as reliable markers for SR resistance. The MLM analysis after the Bonferroni correction did not reveal any associations in this study, although distribution lines in the quantile-quantile (QQ) plot indicates that overcorrection in the test due to both Q and K matrices usage.ConclusionsObtained data suggest that genome wide genotyping of 92 spring barley accessions from Kazakhstan with 9 K Illumina SNP array was highly efficient. Linkage disequilibrium based mapping approach allowed the identification of highly significant QTL for the SR resistance at the HA phase of growth on chromosome 6H. On the other hand, no significant QTL was detected at the SM phase, assuming that for a successful GWASmapping experiment a larger size population with more diverse genetic background should be tested. Obtained results provide additional information towards better understanding of SR resistance in barley.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0686-z) contains supplementary material, which is available to authorized users.

An imputation-based genome-wide association study on traits related to male reproduction in a White Duroc × Erhualian F2 population.

Boar reproductive traits are economically important for the pig industry. Here we conducted a genome-wide association study (GWAS) for 13 reproductive traits measured on 205 F2 boars at day 300 using 60 K single nucleotide polymorphism (SNP) data imputed from a reference panel of 1200 pigs in a White Duroc × Erhualian F2 intercross population. We identified 10 significant loci for seven traits on eight pig chromosomes (SSC). Two loci surpassed the genome-wide significance level, including one for epididymal weight around 60.25 Mb on SSC7 and one for semen temperature around 43.69 Mb on SSC4. Four of the 10 significant loci that we identified were consistent with previously reported quantitative trait loci for boar reproduction traits. We highlighted several interesting candidate genes at these loci, including APN, TEP1, PARP2, SPINK1 and PDE1C. To evaluate the imputation accuracy, we further genotyped nine GWAS top SNPs using PCR restriction fragment length polymorphism or Sanger sequencing. We found an average of 91.44% of genotype concordance, 95.36% of allelic concordance and 0.85 of r(2) correlation between imputed and real genotype data. This indicates that our GWAS mapping results based on imputed SNP data are reliable, providing insights into the genetic basis of boar reproductive traits.

Genetic dissection of blood lipid traits by integrating genome-wide association study and gene expression profiling in a porcine model.

BackgroundSerum concentrations of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and triglycerides (TG) are highly heritable traits that are used clinically to evaluate risk for cardiovascular disease in humans. In this study, we applied a genome-wide association study (GWAS) in 1,075 pigs from two populations and gene expression studies on 497 liver samples to dissect the genetic basis of serum lipids in a pig model.ResultsWe totally identified 8, 5, 2 and 3 genomic loci harboring 109 SNPs that were significantly associated with LDL-C, TC, TG and the ratio of HDL-C/LDL-C in two experimental populations, respectively. In the F2 population, the most prominent SNP was identified at the SSC3: 124,769,847 bp where APOB is the well-known candidate gene. However, in the Sutai population, the most number of significant SNPs was identified at SSC2: 64.97-82.22 Mb where LDLR was identified as the candidate gene. Furthermore, we firstly reported 4 novel genomic loci in pigs harboring the LDL-C-associated SNPs. We also observed obvious population heterogeneity in the two tested populations. Through whole-genome gene expression analysis, we detected 718 trait-correlated expressions. Many of these transcripts correspond to candidate genes for blood lipids in humans. The GWAS mapped 120 cis-eQTLs and 523 trans-eQTLs for these transcripts. One gene encoding the transcript gnl|UG|Ssc#S35330332 stands out to be an important candidate gene for LDL-C by an integrative analysis of GWAS, eQTL and trait-associated expression.ConclusionsWe identified the genomic regions or candidate genes associated with blood lipids by an integrative analysis of GWAS, QTT and eQTL mapping in pigs. The findings would benefit the further identification of the causative genes for blood lipid traits in both pigs and humans.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-14-848) contains supplementary material, which is available to authorized users.

Distinct Variants at LIN28B Influence Growth in Height from Birth to Adulthood

We have studied the largely unknown genetic underpinnings of height growth by using a unique resource of longitudinal childhood height data available in Finnish population cohorts. After applying GWAS mapping of potential genes influencing pubertal height growth followed by further characterization of the genetic effects on complete postnatal growth trajectories, we have identified strong association between variants near LIN28B and pubertal growth (rs7759938; female p = 4.0 x 10(-9), male p = 1.5 x 10(-4), combined p = 5.0 x 10(-11), n = 5038). Analysis of growth during early puberty confirmed an effect on the timing of the growth spurt. Correlated SNPs have previously been implicated as influencing both adult stature and age at menarche, the same alleles associating with taller height and later age of menarche in other studies as with later pubertal growth here. Additionally, a partially correlated LIN28B SNP, rs314277, has been associated previously with final height. Testing both rs7759938 and rs314277 (pairwise r(2) = 0.29) for independent effects on postnatal growth in 8903 subjects indicated that the pubertal timing-associated marker rs7759938 affects prepubertal growth in females (p = 7 x 10(-5)) and final height in males (p = 5 x 10(-4)), whereas rs314277 has sex-specific effects on growth (p for interaction = 0.005) that were distinct from those observed at rs7759938. In conclusion, partially correlated variants at LIN28B tag distinctive, complex, and sex-specific height-growth-regulating effects, influencing the entire period of postnatal growth. These findings imply a critical role for LIN28B in the regulation of human growth.