Marker-trait Associations Research Articles

Insights into maydis leaf blight resistance in maize: a comprehensive genome-wide association study in sub-tropics of India

BackgroundIn the face of contemporary climatic vulnerabilities and escalating global temperatures, the prevalence of maydis leaf blight (MLB) poses a potential threat to maize production. This study endeavours to discern marker-trait associations and elucidate the candidate genes that underlie resistance to MLB in maize by employing a diverse panel comprising 336 lines. The panel was screening for MLB across four environments, employing standard artificial inoculation techniques. Genome-wide association studies (GWAS) and haplotype analysis were conducted utilizing a total of 128,490 SNPs obtained from genotyping-by-sequencing (GBS).ResultsGWAS identified 26 highly significant SNPs associated with MLB resistance, among the markers examined. Seven of these SNPs, reported in novel chromosomal bins (9.06, 5.01, 9.01, 7.04, 4.06, 1.04, and 6.05) were associated with genes: bzip23, NAGS1, CDPK7, aspartic proteinase NEP-2, VQ4, and Wun1, which were characterized for their roles in diminishing fungal activity, fortifying defence mechanisms against necrotrophic pathogens, modulating phyto-hormone signalling, and orchestrating oxidative burst responses. Gene mining approach identified 22 potential candidate genes associated with SNPs due to their functional relevance to resistance against necrotrophic pathogens. Notably, bin 8.06, which hosts five SNPs, showed a connection to defense-regulating genes against MLB, indicating the potential formation of a functional gene cluster that triggers a cascade of reactions against MLB. In silico studies revealed gene expression levels exceeding ten fragments per kilobase million (FPKM) for most genes and demonstrated coexpression among all candidate genes in the coexpression network. Haplotype regression analysis revealed the association of 13 common significant haplotypes at Bonferroni ≤ 0.05. The phenotypic variance explained by these significant haplotypes ranged from low to moderate, suggesting a breeding strategy that combines multiple resistance alleles to enhance resistance to MLB. Additionally, one particular haplotype block (Hap_8.3) was found to consist of two SNPs (S8_152715134, S8_152460815) identified in GWAS with 9.45% variation explained (PVE).ConclusionThe identified SNPs/ haplotypes associated with the trait of interest contribute to the enrichment of allelic diversity and hold direct applicability in Genomics Assisted Breeding for enhancing MLB resistance in maize.

Association mapping with a diverse population of Puccinia graminis f. sp. tritici identified avirulence loci interacting with the barley Rpg1 stem rust resistance gene

BackgroundWheat stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is an important disease of barley and wheat. A diverse sexual Pgt population from the Pacific Northwest (PNW) region of the US contains a high proportion of individuals with virulence on the barley stem rust resistance (R) gene, Rpg1. However, the evolutionary mechanisms of this virulence on Rpg1 are mysterious considering that Rpg1 had not been deployed in the region and the gene had remained remarkably durable in the Midwestern US and prairie provinces of Canada.Methods and resultsTo identify AvrRpg1 effectors, genome wide association studies (GWAS) were performed using 113 Pgt isolates collected from the PNW (n = 89 isolates) and Midwest (n = 24 isolates) regions of the US. Disease phenotype data were generated on two barley lines Morex and the Golden Promise transgenic (H228.2c) that carry the Rpg1 gene. Genotype data was generated by whole genome sequencing (WGS) of 96 isolates (PNW = 89 isolates and Midwest = 7 isolates) and RNA sequencing (RNAseq) data from 17 Midwestern isolates. Utilizing ~1.2 million SNPs generated from WGS and phenotype data (n = 96 isolates) on the transgenic line H228.2c, 53 marker trait associations (MTAs) were identified. Utilizing ~140 K common SNPs generated from combined analysis of WGS and RNAseq data, two significant MTAs were identified using the cv Morex phenotyping data. The 55 MTAs defined two distinct avirulence loci, on supercontig 2.30 and supercontig 2.11 of the Pgt reference genome of Pgt isolate CRL 75-36-700-3. The major avirulence locus designated AvrRpg1A was identified with the GWAS using both barley lines and was delimited to a 35 kb interval on supercontig 2.30 containing four candidate genes (PGTG_10878, PGTG_10884, PGTG_10885, and PGTG_10886). The minor avirulence locus designated AvrRpg1B identified with cv Morex contained a single candidate gene (PGTG_05433). AvrRpg1A haplotype analysis provided strong evidence that a dominant avirulence gene underlies the locus.ConclusionsThe association analysis identified strong candidate AvrRpg1 genes. Further analysis to validate the AvrRpg1 genes will fill knowledge gaps in our understanding of rust effector biology and the evolution and mechanism/s of Pgt virulence on Rpg1.

Insight into a region of chickpea (Cicer arietinum L.) Chromosome 2 revealed potential candidate genes linked to Foc4 Fusarium wilt resistance.

Two markers on Chromosome 2 of chickpea (Cicer arietinum ) are reportedly associated with resistance to race 4 Fusarium wilt, and are frequently used in breeding. However, the genes in this region that actually confer wilt resistance are unknown. We aimed to characterise them using both in silico approaches and marker trait association (MTA) analysis. Of the 225 protein-encoding genes in this region, 51 showed significant differential expression in two contrasting chickpea genotypes under wilt, with potential involvement in stress response. From a diverse set of 244 chickpea genotypes, two sets of 40 resistant and 40 susceptible genotypes were selected based on disease incidence and amplification pattern of the TA59 marker. All cultivars were further genotyped with 1238 single nucleotide polymorphisms (SNPs) specific to the 51 genes; only seven SNPs were significantly correlated with disease. SNP Ca2_24099002, specific to the LOC101498008 (Transmembrane protein 87A) gene, accounted for the highest phenotypic variance for disease incidence at 16.30%, whereas SNPs Ca2_25166118 and Ca2_27029215, specific to the LOC101494644 (β-glucosidase BoGH3B-like) and LOC101505289 (Putative tRNA pseudouridine synthase) genes, explained 10.51% and 10.50% of the variation, respectively, in the sets with contrasting disease susceptibility. Together with the TA59 and TR19 markers, these SNPs can be used in a chickpea breeding scheme to develop wilt resistance.

Chromosome-scale reference genome of an ancient landrace: unveiling the genetic basis of seed weight in the food legume crop pigeonpea (Cajanus cajan).

Pigeonpea (Cajanus cajan) is a nutrient-rich and versatile food legume crop of tropical and subtropical regions. In this study, we describe the de novo assembly of a high-quality genome for the ancient pigeonpea landrace 'D30', achieved through a combination of Pacific Biosciences high-fidelity (PacBio HiFi) and high-throughput chromatin conformation capture (Hi-C) sequencing technologies. The assembled 'D30' genome has a size of 813.54Mb, with a contig N50 of 10.74Mb, a scaffold N50 of 73.07Mb, and a GC content of 35.67%. Genomic evaluation revealed that the 'D30' genome contains 99.2% of Benchmarking Universal Single-Copy Orthologs (BUSCO) and achieves a 29.06 long terminal repeat (LTR) assembly index (LAI). Genome annotation indicated that 'D30' encompasses 431.37Mb of repeat elements (53.02% of the genome) and 37 977 protein-coding genes. Identification of single-nucleotide polymorphisms (SNPs), insertions/deletions (indels), and structural variations between 'D30' and the published genome of pigeonpea cultivar 'Asha' suggests that genes affected by these variations may play important roles in biotic and abiotic stress responses. Further investigation of genomic regions under selection highlights genes enriched in starch and sucrose metabolism, with 42.11% of these genes highly expressed in seeds. Finally, we conducted genome-wide association studies (GWAS) to facilitate the identification of 28 marker-trait associations for six agronomic traits of pigeonpea. Notably, we discovered a calmodulin-like protein (CcCML) that harbors a dominant haplotype associated with the 100-seed weight of pigeonpea. Our study provides a foundational resource for developing genomics-assisted breeding programs in pigeonpea.

Analyzing Medicago spp. seed morphology using GWAS and machine learning

Alfalfa is widely recognized as an important forage crop. To understand the morphological characteristics and genetic basis of seed morphology in alfalfa, we screened 318 Medicago spp., including 244 Medicago sativa subsp. sativa (alfalfa) and 23 other Medicago spp., for seed area size, length, width, length-to-width ratio, perimeter, circularity, the distance between the intersection of length & width (IS) and center of gravity (CG), and seed darkness & red–green–blue (RGB) intensities. The results revealed phenotypic diversity and correlations among the tested accessions. Based on the phenotypic data of M. sativa subsp. sativa, a genome-wide association study (GWAS) was conducted using single nucleotide polymorphisms (SNPs) called against the Medicago truncatula genome. Genes in proximity to associated markers were detected, including CPR1, MON1, a PPR protein, and Wun1(threshold of 1E−04). Machine learning models were utilized to validate GWAS, and identify additional marker-trait associations for potentially complex traits. Marker S7_33375673, upstream of Wun1, was the most important predictor variable for red color intensity and highly important for brightness. Fifty-two markers were identified in coding regions. Along with strong correlations observed between seed morphology traits, these genes will facilitate the process of understanding the genetic basis of seed morphology in Medicago spp.

Genome-wide SNPs and candidate genes underlying the genetic variations for protein and amino acids in pearl millet (Pennisetum glaucum) germplasm

Main conclusionA total of 544 significant marker-trait associations and 286 candidate genes associated with total protein and 18 amino acids were identified. Thirty-three candidate genes were found near the strong marker trait associations (– log10P ≥ 5.5).Pearl millet (Pennisetum glaucum) is largely grown as a subsistence crop in South Asia and sub-Saharan Africa. It serves as a major source of daily protein intake in these regions. Despite its importance, no systematic effort has been made to study the genetic variations of protein and amino acid content in pearl millet germplasm. The present study was undertaken to dissect the global genetic variations of total protein and 18 essential and non-essential amino acids in pearl millet, using a set of 435 K Single Nucleotide Polymorphisms (SNPs) and 161 genotypes of the Pearl Millet Inbred Germplasm Association Panel (PMiGAP). A total of 544 significant marker-trait associations (at P < 0.0001; – log10P ≥ 4) were detected and 23 strong marker-trait associations were identified using Bonferroni’s correction method. Forty-eight pleiotropic loci were found in the genome for the studied traits. In total, 286 candidate genes associated with total protein and 18 amino acids were identified. Thirty-three candidate genes were found near strongly associated SNPs. The associated markers and the candidate genes provide an insight into the genetic architecture of the traits studied and are going to be useful in breeding improved pearl millet varieties in the future. Availabilities of improved pearl millet varieties possessing higher protein and amino acid compositions will help combat the rising malnutrition problem via diet.

Association analysis of leaf aromatic substances in cultivated and weedy types of Perilla crop using SSR markers

In East Asia, particularly South Korea, the two cultivated varieties of Perilla are commonly grown. They are clearly distinguished by their aromatic substances and have different uses as leafy vegetables or oil crop. This study was performed for the development of simple sequence repeat (SSR) markers linked to volatile compounds in Perilla leaves that show differences between cultivated var. frutescens (CF), weedy var. frutescens (WF), and weedy var. crispa (WC) of Perilla. Fifty Perilla SSR primer sets were used to analyze genetic diversity for the 80 Perilla accessions of the three types. A total of 276 alleles were detected, with an average of 5.5 alleles per locus. The average genetic diversity values for CF, WF, and WC accessions were 0.402, 0.583, and 0.437, respectively. WF accessions exhibited the highest genetic diversity among the three types of the Perilla crop. Phylogenetic tree analysis classified 80 Perilla accessions of the three types into four groups, showing 37.2 % genetic similarity. Three types of the Perilla crop were clearly distinguished except for outstanding accessions. Through the application of an association analysis involving 50 Perilla SSR primer sets and five volatile compounds (perilla aldehyde, perilla ketone, myristicin, dill apiol, (Z,E)-α-farnesene) in the three types of the Perilla accessions, we detected 11 significant marker-trait associations duplicated in both Q GLM and Q + K MLM methods. These findings serve as valuable insights for identifying the aromatic substances in Perilla plants originating from various regions of South Korea.

Genome-wide association study reveals genetic loci for ten trace elements in foxtail millet (Setaria italica).

One hundred and fifty-five QTL for trace element concentrations in foxtail millet were identified using a genome-wide association study, and a candidate gene associated with Ni-Co-Cr concentrations was detected. Foxtail millet (Setaria italica) is an important regional crop known for its rich mineral nutrient content, which has beneficial effects on human health. We assessed the concentrations of ten trace elements (Ba, Co, Cr, Cu, Fe, Mn, Ni, Pb, Sr, and Zn) in the grain of 408 foxtail millet accessions. Significant differences in the concentrations of five elements (Ba, Co, Ni, Sr, and Zn) were observed between two subpopulations of spring- and summer-sown foxtail millet varieties. Moreover, 84.4% of the element pairs exhibited significant correlations. To identify the genetic factors influencing trace element accumulation, a comprehensive genome-wide association study was conducted, identifying 155 quantitative trait locus (QTL) for the ten trace elements across three different environments. Among them, ten QTL were consistently detected in multiple environments, including qZn2.1, qZn4.4, qCr4.1, qFe6.3, qFe6.5, qCo6.1, qPb7.3, qPb7.5, qBa9.1, and qNi9.1. Thirteen QTL clusters were detected for multiple elements, which partially explained the correlations between elements. Additionally, the different concentrations of five elements between foxtail millet subpopulations were caused by the different frequencies of high-concentration alleles associated with important marker-trait associations. Haplotype analysis identified a candidate gene SETIT_036676mg associated with Ni accumulation, with the GG haplotype significantly increasing Ni-Co-Cr concentrations in foxtail millet. A cleaved amplified polymorphic sequence marker (cNi6676) based on the two haplotypes of SETIT_036676mg was developed and validated. Results of this study provide valuable reference information for the genetic research and improvement of trace element content in foxtail millet.

Haplotypes of ATP-Binding Cassette CaABCC6 in Chickpea from Kazakhstan Are Associated with Salinity Tolerance and Leaf Necrosis via Oxidative Stress.

Salinity tolerance was studied in chickpea accessions from a germplasm collection and in cultivars from Kazakhstan. After NaCl treatment, significant differences were found between genotypes, which could be arranged into three groups. Those that performed poorest were found in group 1, comprising five ICC accessions with the lowest chlorophyll content, the highest leaf necrosis (LN), Na+ accumulation, malondialdehyde (MDA) content, and a low glutathione ratio GSH/GSSG. Two cultivars, Privo-1 and Tassay, representing group 2, were moderate in these traits, while the best performance was for group 3, containing two other cultivars, Krasnokutsky-123 and Looch, which were found to have mostly green plants and an exact opposite pattern of traits. Marker-trait association (MTA) between 6K DArT markers and four traits (LN, Na+, MDA, and GSH/GSSG) revealed the presence of four possible candidate genes in the chickpea genome that may be associated with the three groups. One gene, ATP-binding cassette, CaABCC6, was selected, and three haplotypes, A, D1, and D2, were identified in plants from the three groups. Two of the most salt-tolerant cultivars from group 3 were found to have haplotype D2 with a novel identified SNP. RT-qPCR analysis confirmed that this gene was strongly expressed after NaCl treatment in the parental- and breeding-line plants of haplotype D2. Mass spectrometry of seed proteins showed a higher accumulation of glutathione reductase and S-transferase, but not peroxidase, in the D2 haplotype. In conclusion, the CaABCC6 gene was hypothesized to be associated with a better response to oxidative stress via glutathione metabolism, while other candidate genes are likely involved in the control of chlorophyll content and Na+ accumulation.

Genotyping-by-sequencing targets genic regions and improves resolution of genome-wide association studies in autotetraploid potato

Key messageDe novo genotyping in potato using methylation-sensitive GBS discovers SNPs largely confined to genic or gene-associated regions and displays enhanced effectiveness in estimating LD decay rates, population structure and detecting GWAS associations over ‘fixed’ SNP genotyping platform. Study also reports the genetic architectures including robust sequence-tagged marker–trait associations for sixteen important potato traits potentially carrying higher transferability across a wider range of germplasm.This study deploys recent advancements in polyploid analytical approaches to perform complex trait analyses in cultivated tetraploid potato. The study employs a ‘fixed’ SNP Infinium array platform and a ‘flexible and open’ genome complexity reduction-based sequencing method (GBS, genotyping-by-sequencing) to perform genome-wide association studies (GWAS) for several key potato traits including the assessment of population structure and linkage disequilibrium (LD) in the studied population. GBS SNPs discovered here were largely confined (~ 90%) to genic or gene-associated regions of the genome demonstrating the utility of using a methylation-sensitive restriction enzyme (PstI) for library construction. As compared to Infinium array SNPs, GBS SNPs displayed enhanced effectiveness in estimating LD decay rates and discriminating population subgroups. GWAS using a combined set of 30,363 SNPs identified 189 unique QTL marker–trait associations (QTL-MTAs) covering all studied traits. The majority of the QTL-MTAs were from GBS SNPs potentially illustrating the effectiveness of marker-dense de novo genotyping platforms in overcoming ascertainment bias and providing a more accurate correction for different levels of relatedness in GWAS models. GWAS also detected QTL ‘hotspots’ for several traits at previously known as well as newly identified genomic locations. Due to the current study exploiting genome-wide genotyping and de novo SNP discovery simultaneously on a large tetraploid panel representing a greater diversity of the cultivated potato gene pool, the reported sequence-tagged MTAs are likely to have higher transferability across a wider range of potato germplasm and increased utility for expediting genomics-assisted breeding for the several complex traits studied.

Screening of CIMMYT and South Asian Bread Wheat Germplasm Reveals Marker-Trait Associations for Seedling Resistance to Septoria Nodorum Blotch.

Wheat (Triticum aestivum L.) production is adversely impacted by Septoria nodorum blotch (SNB), a fungal disease caused by Parastagonospora nodorum. Wheat breeders are constantly up against this biotic challenge as they try to create resistant cultivars. The genome-wide association study (GWAS) has become an efficient tool for identifying molecular markers linked with SNB resistance. This technique is used to acquire an understanding of the genetic basis of resistance and to facilitate marker-assisted selection. In the current study, a total of 174 bread wheat accessions from South Asia and CIMMYT were assessed for SNB reactions at the seedling stage in three greenhouse experiments at CIMMYT, Mexico. The results indicated that 129 genotypes were resistant to SNB, 39 were moderately resistant, and only 6 were moderately susceptible. The Genotyping Illumina Infinium 15K Bead Chip was used, and 11,184 SNP markers were utilized to identify marker-trait associations (MTAs) after filtering. Multiple tests confirmed the existence of significant MTAs on chromosomes 5B, 5A, and 3D, and the ones at Tsn1 on 5B were the most stable and conferred the highest phenotypic variation. The resistant genotypes identified in this study could be cultivated in South Asian countries as a preventative measure against the spread of SNB. This work also identified molecular markers of SNB resistance that could be used in future wheat breeding projects.

Genetic resources and genes/QTLs for gram pod borer (Helicoverpa armigera Hübner) resistance in chickpea from the Western Himalayas.

Helicoverpa armigera (also known as gram pod borer) is a serious threat to chickpea production in the world. A set of 173 chickpea genotypes were evaluated for H. armigera resistance, including mean larval population (MLP), percentage pod damage (PPD), and pest resistance (PR) for 2 consecutive years (year 2020 and 2021). The same core set was also genotyped with 50K AxiomCicerSNP Array. The trait data and 50,000 single nucleotide polymorphism genotypic data were used together to work out marker-trait associations (MTAs) using different genome-wide association studies models. For MLP, a total of 53 MTAs were identified, including 25 MTAs in year 2020 and 28 MTAs in year 2021. A set of three MTAs was found common in both environments. For PPD, two MTAs in year 2020 and five MTAs in year 2021 were identified. A set of two MTAs were common in both environments. Similarly, for PR, only two MTAs common in both environments were identified. Interestingly, a common MTA (Affx_123255526) on chromosome 2 (Ca2) was found to be associated with all the three component traits (MLP, PPD, and PR) of pod borer resistance in chickpea. Further, we report key genes that encode SCAMPs (that facilitates the secretion of defense-related molecules), quinone oxidoreductase (enables the production of reactive oxygen species that promotes diapause of gram pod borer), and NB-LRR proteins that have been implicated in plant defense against H. armigera. The resistant chickpea genotypes, MTAs, and key genes reported in the present study may prove useful in the future for developing pod borer-resistant chickpea varieties.

Genome-wide association study reveals marker-trait associations for major agronomic traits in proso millet (Panicum miliaceum L.).

The pilot-scale genome-wide association study in the US proso millet identified twenty marker-trait associations for five morpho-agronomic traits identifying genomic regions for future studies (e.g. molecular breeding and map-based cloning). Proso millet (Panicum miliaceum L.) is an ancient grain recognized for its excellent water-use efficiency and short growing season. It is an indispensable part of the winterwheat-based dryland cropping system in the High Plains of the USA. Its grains are endowed with high nutritional and health-promoting properties, making it increasingly popular in the global market for healthy grains. There is a dearth of genomic resources in proso millet for developing molecular tools to complement conventional breeding for developing high-yielding varieties. Genome-wide association study (GWAS) is a widely used method to dissect the genetics of complex traits. In this pilot study of the first-ever GWAS in the US proso millet, 71 globally diverse genotypes of 109 the US proso millet core collection were evaluated for five major morpho-agronomic traits at two locations in western Nebraska, and GWAS was conducted to identify single nucleotide polymorphisms (SNPs) associated with these traits. Analysis of variance showed that there was a significant difference among the genotypes, and all five traits were also found to be highly correlated with each other. Sequence reads from genotyping-by-sequencing (GBS) were used to identify 11,147 high-quality bi-allelic SNPs. Population structure analysis with those SNPs showed stratification within the core collection. The GWAS identified twenty marker-trait associations (MTAs) for the five traits. Twenty-nine putative candidate genes associated with the five traits were also identified. These genomic regions can be used to develop genetic markers for marker-assisted selection in proso millet breeding.

Genome-wide association and genomic prediction study of elite spring bread wheat (Triticum aestivum L.) genotypes under drought conditions across different locations

Abiotic stress, notably drought, impacts wheat production globally, but more so in central and South Asia, North Africa (CWANA), and sub-Saharan Africa (SSA). The current study attempts to identify significant markers linked to drought and heat tolerance and assess genomic prediction. A genome-wide association study was conducted using the 10 K wheat SNP markers for grain yield and related traits of 246 spring bread wheat genotypes from ICARDA. Traits including grain yield (GY), days to heading (DHE), days to maturity (DMA), plant height (PLH), and thousand kernel weight (TKW), were evaluated across six different locations, spanning two years 2015–2016 and 2016–2017, as per variance analysis. Grain yield and related-traits showed a considerable variation among genotypes. Moreover, GWAS using a mixed linear model (MLM), revealed 65 marker-trait associations (MTAs) across the six environments on 16 chromosomes. With an average r2 value of 0.26, Genome D has the highest linkage, followed by Genomes B and A with r2 values of 0.22 and 0.21, respectively. GY had the highest MTA rating (35), followed by TKW (9) and 3 for each of the other agronomic traits (DHE, DMA, PLH) at Merchouch station. The marker “CAP8_c1393_327” was the most significant associated marker correlated with grain yield located on chromosome 3 A across Sid El Aidi station. Additionally, the SNP markers “wsnp_Ra_c26091_35652620” displayed extremely significant and stable MTA for TKW on chromosome 5B at Merchouch station. The markers and candidate genes reported throughout this study have the potential to be used in marker-assisted selection to enhance wheat genotypes in terms of yield and resistance to drought limitations.

Genetic dissection of minerals and phytate content in pearl millet germplasm panel using genome‐wide association study

AbstractBillions of people around the world suffer from malnutrition, leading to severe adverse health effects. Pearl millet (Pennisetum glaucum) is a multifaceted versatile crop with excellent nutritional profile which can help to combat nutritional disorders and climate change. In this study, we evaluated the global pearl millet germplasm panel known as PMiGAP for natural variation and genetic maker trait associations for important minerals, that is, iron, zinc, calcium, magnesium, potassium and sodium along with phytate. The genotypes IP‐15947, IP‐5121, IP‐4020, IP‐12768, IP‐5695, IP‐8786 and IP‐11310 were found to be superior for majority of minerals examined but had lower phytate‐to‐zinc ratio. Phytate/mineral molar ratios are typically used to predict the bioavailability of iron and calcium contents in grains, and surprisingly none of the PMiGAP genotypes showed such ratios below threshold indicating PMiGAP entries studied in this study seriously suffer from bioavailability issues of these minerals. On the other hand, 73 genotypes had lower zinc/phytate ratio than the threshold in the germplasm panel. Iron and zinc content had significant positive association among them but phytate content in general was not significantly correlated with minerals except for magnesium and potassium. A genome‐wide association study using 456 K SNPs identified 74 significant marker–trait associations and 59 candidate genes around 50 Kb distance near the significant SNPs. Ten significant SNPs were found within the candidate genes. The associated markers and the candidate genes provide new insights into the genetic architecture of the mineral traits studied and will facilitate marker‐assisted selection to accelerate breeding of such minerals in future varieties to combat rising malnutrition problem via diet.

Capitalizing on genebank core collections for rare and novel disease resistance loci to enhance barley resilience.

In the realm of agricultural sustainability, the utilization of plant genetic resources for enhanced disease resistance is paramount. Preservation efforts in genebanks are justified by their potential contributions to future crop improvement. To capitalize on the potential of plant genetic resources, we focused on a barley core collection from the German ex situ genebank and contrasted it with a European elite collection. The phenotypic assessment included 812 plant genetic resources and 298 elites, with a particular emphasis on four disease traits (Puccinia hordei, Blumeria graminis hordei, Ramularia collo-cygni, and Rhynchosporium commune). An integrated genome-wide association study, employing both Bayesian-information and linkage-disequilibrium iteratively nested keyway (BLINK) and a linear mixed model, was performed to unravel the genetic underpinnings of disease resistance. A total of 932 marker-trait associations were identified and assigned to 49 quantitative trait loci. The accumulation of novel and rare resistance alleles significantly bolstered the overall resistance level in plant genetic resources. Three plant genetic resources donors with high counts of novel/rare alleles and exhibiting exceptional resistance to leaf rust and powdery mildew were identified, offering promise for targeted pre-breeding goals and enhanced resilience in future varieties. Our findings underscore the critical contribution of plant genetic resources to strengthening crop resilience and advancing sustainable agricultural practices.

Multi-model genome-wide association study on key organic naked barley agronomic, phenological, diseases, and grain quality traits

The study objective was to assess the potential benefits of using genomic tools in organic plant breeding programs to enhance selection efficiency. A diversity panel of 247 spring naked barley accessions was characterized under Irish organic conditions over 3 years. Genome-wide association studies (GWAS) were performed on 19 traits related to agronomy, phenology, diseases, and grain quality, using the information on 50 K Single Nucleotide Polymorphisms (SNP). Four models (EMMA, G model, BLINK, 3VMrMLM) were applied to 5 types of Best Linear Unbiased Predictors (BLUP): within-year, mean, aggregated within-year). 1653 Marker-Trait-Associations (MTA) were identified, with 259 discovered in at least two analyses. 3VMrMLM was the best-performing model with significant MTA together explaining the largest proportion of the additive variance for most traits and BLUP types (from 1.4 to 50%). This study proposed a methodology to prioritize main effect MTA from different models’ outputs, using multi-marker regression analyses with markers fitted as fixed or random factors. 36 QTL, considered major, explained more than 5% of the trait variance on each BLUP type. A candidate gene or known QTL was found for 18 of them, with 13 discovered with 3VMrMLM. Multi-model GWAS was useful for validating additional QTL, including 8 only discovered with BLINK or G model, thus allowing a broader understanding of the traits’ genetic architecture. In addition, results highlighted a correlation between the trait value and the number of favorable major QTL exhibited by accessions. We suggest inputting this number in a multi-trait index for a more efficient Marker-Assisted Selection (MAS) of accessions best balancing multiple quantitative traits.

Identification of novel marker-trait associations and candidate genes for combined low phosphorus and nitrogen-deficient conditions in rice at seedling stage

Identification of novel marker-trait associations and candidate genes for combined low phosphorus and nitrogen-deficient conditions in rice at seedling stage

Quantitative Trait Loci Mapping and Association Analysis of Solanesol Content in Tobacco (Nicotiana tabacum L.)

Solanesol, which accumulates predominantly in the leaves of tobacco plants, has medically important bioactive properties. To investigate the genetic basis of solanesol in tobacco (Nicotiana tabacum), the solanesol contents of 222 accessions, 206 individuals from an N. tabacum Maryland609 (low-solanesol) × K326 (high-solanesol) F2 population and their corresponding F1 self-pollinations, were determined using ultra-performance liquid chromatography. Genome-wide quantitative trait locus (QTL) and association analysis were performed to identify QTLs and markers associated with solanesol content based on simple sequence repeat molecular markers. A total of 12 QTLs underlying solanesol content were mapped to seven linkage groups (LGs), with three of the QTLs (QTL3-1, QTL21-6, and QTL23-3) explaining 5.19–10.05% of the phenotypic variation. Association analysis revealed 38 significant marker-trait associations in at least one environment. The associations confirmed the QTLs located on LG3, LG10, LG14, LG21, and LG23, while new elite makers were located on 11 additional LGs, each explaining, respectively, 5.16–20.07% of the phenotypic variation. The markers LG14-PT54448, LG10-PT60114-2, LG10-PT60510, LG10-PT61061, and LG-21PT20388 may be useful for molecular-assisted selection of solanesol content in tobacco leaves. These results increase our understanding of the inheritance of solanesol-associated genes and will contribute to molecular-assisted breeding and further isolation of regulatory genes involved in solanesol biosynthesis in tobacco leaves.

Linkage Mapping and Genome-Wide Association Study Identified Two Peanut Late Leaf Spot Resistance Loci, PLLSR-1 and PLLSR-2, Using Nested Association Mapping.

Identification of candidate genes and molecular markers for late leaf spot (LLS) disease resistance in peanut (Arachis hypogaea) has been a focus of molecular breeding for the U.S. industry-funded peanut genome project. Efforts have been hindered by limited mapping resolution due to low levels of genetic recombination and marker density available in traditional biparental mapping populations. To address this, a multi-parental nested association mapping population has been genotyped with the peanut 58K single-nucleotide polymorphism (SNP) array and phenotyped for LLS severity in the field for 3 years. Joint linkage-based quantitative trait locus (QTL) mapping identified nine QTLs for LLS resistance with significant phenotypic variance explained up to 47.7%. A genome-wide association study identified 13 SNPs consistently associated with LLS resistance. Two genomic regions harboring the consistent QTLs and SNPs were identified from 1,336 to 1,520 kb (184 kb) on chromosome B02 and from 1,026.9 to 1,793.2 kb (767 kb) on chromosome B03, designated as peanut LLS resistance loci, PLLSR-1 and PLLSR-2, respectively. PLLSR-1 contains 10 nucleotide-binding site leucine-rich repeat disease resistance genes. A nucleotide-binding site leucine-rich repeat disease resistance gene, Arahy.VKVT6A, was also identified on homoeologous chromosome A02. PLLSR-2 contains five significant SNPs associated with five different genes encoding callose synthase, pollen defective in guidance protein, pentatricopeptide repeat, acyl-activating enzyme, and C2 GRAM domains-containing protein. This study highlights the power of multi-parent populations such as nested association mapping for genetic mapping and marker-trait association studies in peanuts. Validation of these two LLS resistance loci will be needed for marker-assisted breeding.