Single Nucleotide Polymorphism Map Research Articles

Abstract P42: Genetic Variants Associated With Vasospasm Following Aneurysmal Subarachnoid Hemorrhage

Background: Cerebral vasospasm is a major contributor to poor clinical outcomes in intracranial aneurysm (IA) patients and is involved in delayed cerebral ischemia (DCI) after aneurysmal subarachnoid hemorrhage (aSAH). Little is known about the genetic factors associated with vasospasm pathogenesis. We performed a genome-wide association study (GWAS) to investigate association of common genetic variants with risk of vasospasm in IA patients with SAH. Methods: Our Caucasian cohort of 190 aSAH patients (121 with vasospasm diagnosed by angiography) were genotyped using UK Biobank Axiom Arrays. Genotypes were called using the Best Practices Workflow in the Axiom Analysis Suite v4.0.3.3. All samples had >97% call rate, one sample was removed due to sex mismatch, and a total of 634,781 single nucleotide polymorphisms (SNPs) with >0.01 minor allele frequency were analyzed for association with vasospasm in the final cohort of 189 cases (66.7% female, mean age 54.8 years). Logistic regression analysis was performed using an additive model adjusting for age at onset of symptoms and sex. Genome-wide significance was based on Bonferroni correction for multiple testing (P< 7.9x10 -8 ). Results: No SNPs were significantly associated with vasospasm at the genome-wide level. However, eleven SNPs were nominally associated with vasospasm (P<10E-5) including the most significant SNP mapping to an intron of GLIS3 (P=9.74E-06, OR=3.94, 95% CI: 2.15-7.22), three intronic SNPs mapping to ACVR1 , and additional SNPs mapping to introns in FSTL3 , TMEM242 , DCLK1 , one SNP upstream of RAC2 and two SNPs downstream of TEX29 and TGFBR3 , and one missense SNP mapping to MYO18B. Interestingly, two of these genes ( ACVR1 and TGFBR3 ) are in the transforming growth factor-beta (TGF-beta) signaling pathway, previously implicated in SAH pathogenesis. Conclusions: We performed a GWAS of vasospasm following aSAH. Our study suggests that common SNPs may contribute to vasospasm susceptibility following aSAH and suggests a potential role for TGF-beta signaling pathway genes in vasospasm. These findings warrant further replication in additional independent studies including other DCI outcomes.

Characterization of wheat curl mite resistance gene Cmc4 in OK05312

Cmc4, a wheat curl mite resistance gene, was delimited to a 523 kb region and a diagnostic marker haplotype was identified for selecting Cmc4 in breeding programs. Wheat curl mite (WCM, Aceria tosichella Keifer) is a disastrous wheat pest in many wheat-growing regions worldwide. WCM not only directly affects wheat yield, but also transmits wheat streak mosaic virus. Growing WCM resistant cultivars is the most economical and sustainable method to reduce its damage. A hard winter wheat breeding line OK05312 (PI 670019) carries Cmc4 gene resistance to A. tosichella and has many desirable agronomic traits. To finely map Cmc4 in OK05312, two recombinant inbred line populations were developed from crosses between OK05312 and two susceptible cultivars, SD06165 and Jerry, genotyped using single nucleotide polymorphism (SNP) markers generated from genotyping-by-sequencing (GBS), and phenotyped for WCM resistance. Gene mapping using the two SNP maps confirmed Cmc4 in OK05312 that explained up to 68% of the phenotypic variation. Further analysis delimited Cmc4 to a ~ 523kb region between SNPs SDOKSNP6314 and SDOKSNP2805 based on the Ae. tauschii reference genome. We developed 18 polymorphic Kompetitive Allele Specific PCR (KASP) markers using the sequences of GBS-SNPs in this region and 23 additional KASP markers based on the SNPs between the parents derived from 90K SNP chips. The KASP markers SDOKSNP6314 and SDOKSNP9699 are closest to Cmc4 and can be used to diagnose the presence of Cmc4 in wheat breeding programs. Haplotype analysis suggested that CmcTAM112 in TAM112 might be the same gene as Cmc4.

Identification of Promising RILs for High Grain Zinc Through Genotype × Environment Analysis and Stable Grain Zinc QTL Using SSRs and SNPs in Rice (Oryza sativa L.).

Polished rice is one of the commonly consumed staple foods across the world. However, it contains limited nutrients especially iron (Fe) and zinc (Zn). To identify promising recombinant inbred lines (RILs) for grain Zn and single plant yield, 190 RILs developed from PR116 and Ranbir Basmati were evaluated in two environments (E1 and E2). A subset of 44 contrasting RILs for grain Zn was screened in another two environments (E3 and E4). Phenotypic data was collected for 10 traits, viz., days to 50% flowering, plant height, panicle length, number of tillers, single plant yield (SPY), test weight, Fe and Zn in brown (IBR, ZBR), and polished rice (IPR, ZPR). Stepwise regression analysis of trait data in 190 RILs and a subset of 44 RILs revealed the interdependence of ZPR, ZBR, IPR, and IBR and the negative association of grain Zn with single plant yield. Based on the additive main effect and multiplicative interaction (AMMI) and genotype and genotype × environment interaction (GGE) analyses of the subset of 44 RILs across four environments (E1–E4), six promising RILs were identified for ZPR with >28 ppm. Mapping of 190 RILs with 102 simple sequence repeats (SSRs) resulted in 13 QTLs for best linear unbiased estimates (BLUEs) of traits including advantage over check (AOC). Using genotype-based sequencing (GBS), the subset of 44 RILs was mapped with 1035 single-nucleotide polymorphisms (SNPs) and 21 QTLs were identified. More than 100 epistatic interactions were observed. A major QTL qZPR.1.1 (PV 37.84%) and another QTL qZPR.11.1 (PV 15.47%) were identified for grain Zn in polished rice. A common major QTL (qZBR.2.1 and qZPR.2.1) was also identified on chromosome 2 for grain Zn content across SSR and SNP maps. Two potential candidate genes related to transporters were identified based on network analyses in the genomic regions of QTL < 3 Mb. The RILs identified for grain Zn and SPY were nominated for national evaluation as under rice biofortification, and two QTLs identified based on BLUEs could be used in the rice biofortification breeding programs.

A Major QTL for Resistance to Vibrio anguillarum in Rainbow Trout.

Genetic selection of disease resistant fish is a major strategy to improve health, welfare and sustainability in aquaculture. Mapping of single nucleotide polymorphisms (SNP) in the fish genome may be a fruitful tool to define relevant quantitative trait loci (QTL) and we here show its use for characterization of Vibrio anguillarum resistant rainbow trout (Oncorhynchus mykiss). Fingerlings were exposed to the pathogen V. anguillarum serotype O1 in a solution of 1.5 × 107 cfu/ml and observed for 14 days. Disease signs appeared 3 days post exposure (dpe) whereafter mortality progressed exponentially until 6 dpe reaching a total mortality of 55% within 11 days. DNA was sampled from all fish – including survivors – and analyzed on a 57 k Affymetrix SNP platform whereby it was shown that disease resistance was associated with a major QTL on chromosome 21 (Omy 21). Gene expression analyses showed that diseased fish activated genes associated with innate and adaptive immune responses. The possible genes associated with resistance are discussed.

TSETA: A Third-Generation Sequencing-Based Computational Tool for Mapping and Visualization of SNPs, Meiotic Recombination Products, and RIP Mutations.

TSETA (Third-generation Sequencing to Enable Tetrad Analysis) is a fungus-centric software pipeline that utilizes chromosome-level sequence assembly for genome-wide and single-nucleotide-resolution mapping of single-nucleotide polymorphisms (SNPs), meiotic recombination products, illegitimatemutations (IMs)and repeat-induced point (RIP) mutations. It utilizes a newly invented algorithm (i.e., BLASTN-guided sectional MAFFT) to perform fast, accurate, and low-cost multiple genome sequence alignments. This new algorithm outcompetes next-generation sequencing (NGS)-based variant-calling approaches for accurate and comprehensive identification of single-nucleotide variants (SNVs) and insertion/deletion mutations (Indels) among the near-complete genome sequences of any two or more intraspecific strains, as well as sequences before and after meiosis, with single-nucleotide precision. TSETA also has a powerful tool for the visualization of the results from the scale of the chromosomal landscape to individual nucleotides. The data output files are user-friendly for researchers and students lacking computational expertise to analyze and reason about data and evidence.

Genome-Wide SNP Discovery and Mapping QTLs for Seed Iron and Zinc Concentrations in Chickpea (Cicer arietinum L.).

Biofortification through plant breeding is a cost-effective and sustainable approach towards addressing micronutrient malnutrition prevailing across the globe. Screening cultivars for micronutrient content and identification of quantitative trait loci (QTLs)/genes and markers help in the development of biofortified varieties in chickpea (Cicer arietinum L.). With the aim of identifying the genomic regions controlling seed Fe and Zn concentrations, the F2:3 population derived from a cross between MNK-1 and Annigeri 1 was genotyped using genotyping by sequencing approach and evaluated for Fe and Zn concentration. An intraspecific genetic linkage map comprising 839 single nucleotide polymorphisms (SNPs) spanning a total distance of 1,088.04 cM with an average marker density of 1.30 cM was constructed. By integrating the linkage map data with the phenotypic data of the F2:3 population, a total of 11 QTLs were detected for seed Fe concentration on CaLG03, CaLG04, and CaLG05, with phenotypic variation explained ranging from 7.2% (CaqFe3.4) to 13.4% (CaqFe4.2). For seed Zn concentration, eight QTLs were identified on CaLG04, CaLG05, and CaLG08. The QTLs individually explained phenotypic variations ranging between 5.7% (CaqZn8.1) and 13.7% (CaqZn4.3). Three QTLs for seed Fe and Zn concentrations (CaqFe4.4, CaqFe4.5, and CaqZn4.1) were colocated in the “QTL-hotspot” region on CaLG04 that harbors several drought tolerance-related QTLs. We identified genes in the QTL regions that encode iron–sulfur metabolism and zinc-dependent alcohol dehydrogenase activity on CaLG03, iron ion binding oxidoreductase on CaLG04, and zinc-induced facilitator-like protein and ZIP zinc/iron transport family protein on CaLG05. These genomic regions and the associated markers can be used in marker-assisted selection to increase seed Fe and Zn concentrations in agronomically superior chickpea varieties.

Genomic Dissection of a Wild Region in a Superior Solanum pennellii Introgression Sub-Line with High Ascorbic Acid Accumulation in Tomato Fruit.

The Solanum pennellii introgression lines (ILs) have been exploited to map quantitative trait loci (QTLs) and identify favorable alleles that could improve fruit quality traits in tomato varieties. Over the past few years, ILs exhibiting increased content of ascorbic acid in the fruit have been selected, among which the sub-line R182. The aims of this work were to identify the genes of the wild donor S. pennellii harbored by the sub-line and to detect genes controlling ascorbic acid accumulation by using genomics tools. A Genotyping-By-Sequencing (GBS) approach confirmed that no wild introgressions were present in the sub-line besides one region on chromosome 7. By using a dense single nucleotide polymorphism (SNP) map obtained by RNA sequencing (RNA-Seq), the wild region of the sub-line was finely identified; thus, defining 39 wild genes that replaced 33 genes of the ILs genetic background (cv. M82). The differentially expressed genes mapping in the region and the variants detected among the cultivated and the wild alleles evidenced the potential role of the novel genes present in the wild region. Interestingly, one upregulated gene, annotated as a major facilitator superfamily protein, showed a novel structure in R182, with respect to the parental lines. These genes will be further investigated using gene editing strategies.

Genome-wide association studies reveal QTL hotspots for grain brightness and black point traits in barley

Grain kernel discoloration (KD) in cereal crops leads to down-grading grain quality and substantial economic losses worldwide. Breeding KD tolerant varieties requires a clear understanding of the genetic basis underlying this trait. Here, we generated a high-density single nucleotide polymorphisms (SNPs) map for a diverse barley germplasm and collected trait data from two independent field trials for five KD related traits: grain brightness (TL), redness (Ta), yellowness (Tb), black point impact (Tbpi), and total black point in percentage (Tbpt). Although grain brightness and black point is genetically correlated, the grain brightness traits (TL, Ta, and Tb) have significantly higher heritability than that of the black point traits (Tbpt and Tbpi), suggesting black point traits may be more susceptible to environmental influence. Using genome-wide association studies (GWAS), we identified a total of 37 quantitative trait loci (QTL), including two major QTL hotspots on chromosomes 4H and 7H, respectively. The two QTL hotspots are associated with all five KD traits. Further genetic linkage and gene transcription analyses identified candidate genes for the grain KD, including several genes in the flavonoid pathway and plant peroxidase. Our study provides valuable insights into the genetic basis for the grain KD in barley and would greatly facilitate future breeding programs for improving grain KD resistance.

Low Additive Genetic Variation in a Trait Under Selection in Domesticated Rice.

Quantitative traits are important targets of both natural and artificial selection. The genetic architecture of these traits and its change during the adaptive process is thus of fundamental interest. The fate of the additive effects of variants underlying a trait receives particular attention because they constitute the genetic variation component that is transferred from parents to offspring and thus governs the response to selection. While estimation of this component of phenotypic variation is challenging, the increasing availability of dense molecular markers puts it within reach. Inbred plant species offer an additional advantage because phenotypes of genetically identical individuals can be measured in replicate. This makes it possible to estimate marker effects separately from the contribution of the genetic background not captured by genotyped loci. We focused on root growth in domesticated rice, Oryza sativa, under normal and aluminum (Al) stress conditions, a trait under recent selection because it correlates with survival under drought. A dense single nucleotide polymorphism (SNP) map is available for all accessions studied. Taking advantage of this map and a set of Bayesian models, we assessed additive marker effects. While total genetic variation accounted for a large proportion of phenotypic variance, marker effects contributed little information, particularly in the Al-tolerant tropical japonica population of rice. We were unable to identify any loci associated with root growth in this population. Models estimating the aggregate effects of all measured genotypes likewise produced low estimates of marker heritability and were unable to predict total genetic values accurately. Our results support the long-standing conjecture that additive genetic variation is depleted in traits under selection. We further provide evidence that this depletion is due to the prevalence of low-frequency alleles that underlie the trait.

Identification and fine mapping of spot blotch (Bipolaris sorokiniana) resistance gene Sb4 in wheat.

A new spot blotch (Bipolaris sorokiniana) resistance gene Sb4 was mapped in a genomic interval of 1.34Mb on wheat chromosome 4BL. Spot blotch, caused by Bipolaris sorokiniana, has emerged as a serious concern for cultivation of wheat in warmer and humid regions of the world, which results in substantial yield losses and descends with quality. In this study, we identified and mapped a spot blotch resistance gene, designated as Sb4, against B. sorokiniana in wheat. Bulked segregant RNA-Seq (BSR-Seq) analysis and single-nucleotide polymorphism mapping showed that Sb4 is located on the long arm of chromosome 4B. A genetic linkage map of Sb4 was constructed using an F4 mapping population developed from the cross between 'GY17' and 'Zhongyu1211,' and Sb4 was delimited in a 7.14-cM genetic region on 4BL between markers B6811 and B6901. Using the Chinese Spring reference sequences of chromosome arm 4BL, 13 new polymorphic markers were developed. Finally, Sb4 was mapped in a 1.19-cM genetic interval corresponding to a 1.34-Mb physical genomic region of Chinese Spring chromosome 4BL containing 21 predicted genes. This study provides a foundational step for further cloning of Sb4 using a map-based approach.

Reassigning Hessian fly resistance genes H7 and H8 to chromosomes 6A and 2B of the wheat cultivar ‘Seneca’ using genotyping‐by‐sequencing

AbstractHessian fly (HF, Mayetiola destructor Say), is a destructive insect pest in most wheat (Triticum aestivum L.) growing areas worldwide. Growing resistant cultivars can effectively reduce HF damage. The wheat cultivar ‘Seneca’ was reported to carry resistance genes H7 on chromosome 5D and H8 with unknown location. To further validate the resistance gene locations in Seneca, a recombinant inbred line (RIL) population from Bobwhite × Seneca was genotyped using 3,330 single nucleotide polymorphism (SNP) markers generated from genotyping‐by‐sequencing (GBS) and phenotyped for HF resistance in the greenhouse experiments. Phenotypic analyses showed at least two genes conditioning HF resistance in Seneca, but none of them was mapped on chromosome 5D, as previously reported. Instead, one major gene, designated as H7, was mapped to chromosome 6A and explained 60.7–78.3% of the phenotypic variation. The other gene with a minor effect, designated as H8, was found on chromosome 2B and explained 3.2–4.7% of the phenotypic variation. Based on the physical locations of flanking markers for H7 in the Chinese Spring reference genome, H7 was physically located in a 6‐Mb interval on 6AL. This study reassigned the major gene H7 to chromosome 6A and the minor gene H8 to 2B in Seneca using a high‐density SNP map. Seventeen GBS SNPs mapped in the H7 region were converted into Kompetitive allele specific polymerase chain reaction (KASP) markers for selecting this gene in breeding.

Association of IL13 genetic polymorphisms with atopic dermatitis: Fine mapping and haplotype analysis

BackgroundAlthough previous studies had reported an important role of interleukin 13 (IL13) and its genetic polymorphisms in atopic dermatitis (AD), many of these previous reports focused on the missense variant rs20541 (Gln144Arg) without fine mapping of the gene region. ObjectiveTo analyze the potential associations of other IL13 variants and their haplotypes with AD and assess total serum immunoglobulin E (IgE) levels. MethodsWe performed fine mapping of single-nucleotide polymorphisms (SNPs) within the IL13 gene in a pilot study of 495 children with AD and 444 healthy controls. Then, we conducted a replication study of 757 children with AD and 1620 healthy controls to evaluate the association between the rs20541 variant of IL13 and AD. ResultsIn the pilot study, the rs20541 and rs1295685 SNPs in the 3ʹ-untranslated region of IL13 had significant associations with AD (P < .001 and .01, respectively). In addition, 2 haplotypes (BL2_ht1 and BL2_ht2), which harbored the significant rs20541 and rs1295685 SNPs, had an association with AD (minimum P = .006). BL2_ht1 and BL2_ht2 had nominal signals associated with the total serum IgE levels (P < .05) but not with the severity of AD (P > .05). In the replication study, rs20541 was associated with the total serum IgE levels but not with the severity of AD. ConclusionAn additional IL13 gene SNP, rs1295685, has a strong linkage disequilibrium with rs20541, and its haplotypes are associated with AD and the total serum IgE levels.

Optimization of Genotype by Sequencing data for phylogenetic purposes

• Herein we propose a framework for assembling and analyzing Genotype by Sequencing (GBS) data to better understand evolutionary relationships within a group of closely related species using the mastiff bats (Molossus) as our model system. Many species within this genus have low-levels of genetic variation within and between morphologically distinct species, and the relationships among them remain unresolved using traditional Sanger sequencing methods. Given that both de novo and reference genome pipelines can be used to assemble next generation sequences, and that several tree inference methodologies have been proposed for single nucleotide polymorphism (SNP) data, we test whether different alignments and phylogenetic approaches produce similar results. We also examined how the process of SNP identification and mapping can affect the consistency of the analyses. Different alignments and phylogenetic inferences produced consistent results, supporting the GBS approach for answering evolutionary questions on a macroevolutionary scale when the genetic distance among phenotypically identifiable clades is low. We highlight the importance of exploring the relationships among groups using different assembly assumptions and also distinct phylogenetic inference methods, particularly when addressing phylogenetic questions in genetic and morphologically conservative taxa.• The method uses the comparison of several filter settings, alignments, and tree inference approaches on Genotype by Sequencing data.• Consistent results were found among several approaches.• The methodology successfully recovered well supported species boundaries and phylogenetic relationships among species of mastiff bats not hypothesized by previous methods.

Single nucleotide polymorphisms (SNPs) provide unprecedented resolution of species boundaries, phylogenetic relationships, and genetic diversity in the mastiff bats (Molossus)

Mammals are one of the better known groups of animals, and in the Neotropics bats typically comprise about half of the mammalian species diversity. But, well resolved species-level phylogenies are still lacking for most taxa of bats. One broadly distributed genus is the mastiff bats, Molossus. Species within this genus are morphologically very similar, which results in a confusing and unstable taxonomy. In addition, low levels of genetic divergence among some clades make resolution of phylogenetic relationships difficult. Most authors recognize Molossus as being monophyletic, however, phylogenetic relationships within the genus remain poorly understood based on traditional Sanger sequencing of individual genes. We propose a more comprehensive framework based on large-scale genomic data derived from Next Generation Sequencing techniques to better understand evolutionary relationships within a group of closely related species with a rich taxonomic history. In this study, we utilized the NGS method of Genotype by Sequencing (GBS) to test the monophyly of the genus, understand evolutionary relationships within Molossus and investigate the genetic integrity of currently recognized species. Given that both de novo and reference genome pipelines are often used in the assembly of Single Nucleotide Polymorphism data from GBS, and that several tree inference methodologies have been proposed for SNP data, we test whether different alignments and phylogenetic approaches produce similar results. We also examined how the process of SNP identification and mapping can affect the consistency of the analyses. Our data provide the first high resolution phylogeny for the genus Molossus, bringing new insights into recognition of species boundaries and relationships among taxa. This study clarifies the taxonomy of Molossus and elevates the number of species in the genus from 11 to 14. We suggest the revalidation of the names M. nigricans, and M. fluminensis, which were synonymized under the name M. rufus; and M. bondae, previously synonymized under the name M. currentium. Different alignments and phylogenetic inferences produce consistent results, supporting use of SNP approach in addressing evolutionary questions on a macroevolutionary scale where the genetic distance among clades is low.

Cellulose defects in the Arabidopsis secondary cell wall promote early chloroplast development.

Lincomycin (LIN)-mediated inhibition of protein synthesis in chloroplasts prevents the greening of seedlings, represses the activity of photosynthesis-related genes in the nucleus, including LHCB1.2, and induces the phenylpropanoid pathway, resulting in the production of anthocyanins. In genomes uncoupled (gun) mutants, LHCB1.2 expression is maintained in the presence of LIN or other inhibitors of early chloroplast development. In a screen using concentrations of LIN lower than those employed to isolate gun mutants, we have identified happy on lincomycin (holi) mutants. Several holi mutants show an increased tolerance to LIN, exhibiting de-repressed LHCB1.2 expression and chlorophyll synthesis in seedlings. The mutations responsible were identified by whole-genome single-nucleotide polymorphism (SNP) mapping, and most were found to affect the phenylpropanoid pathway; however, LHCB1.2 expression does not appear to be directly regulated by phenylpropanoids, as indicated by the metabolic profiling of mutants. The most potent holi mutant is defective in a subunit of cellulose synthase encoded by IRREGULAR XYLEM3, and comparative analysis of this and other cell-wall mutants establishes a link between secondary cell-wall integrity and early chloroplast development, possibly involving altered ABA metabolism or sensing.

28. KARYOMAPPING USING WHOLE GENOME SEQUENCING DATA: PRECLINICAL VALIDATION OF A PERSONALISED APPROACH TO PGT-M

Introduction Karyomapping utilises microarray technology to track informative single nucleotide polymorphisms (SNPs) and assign haplotypes to embryos without the need for direct mutation detection (Handyside et al. 2010). Although suitable for most couples undergoing PGT-M, the use of a fixed set of SNPs may provide inadequate information in certain genetic regions or in consanguineous couples (Konstantinidis et al. 2015). Whole genome sequencing (WGS) has the potential to individualise informative SNP detection and perform concurrent direct mutation detection. However, no studies have evaluated the potential for application of the karyomapping algorithm to WGS data. Material and Methods The Platinum Genomes dataset ( https://sapac.illumina.com/platinumgenomes.html ) was analysed as a model for the application of karyomapping to WGS data. Variant call format (VCF) files were analysed according to the principles of karyomapping to create a map of informative biallelic SNPs in each grandparental haplotype. A model of embryo biopsy was created by diluting DNA from one of the family members (NA12878) to 30pg and whole genome amplification was performed with the Repli-G single cell kit (Qiagen, Hilden, Germany) as a model of trophectoderm biopsy. Whole genome sequencing was performed in triplicate on a high output NextSeq 2500 run (Illumina, California, USA) at average 15x coverage and the number of informative SNPs detected compared to the karyomapping SNP microarray. Comparison of variant calling methods was performed using the Genome Analysis Toolkit v3.4 (Broad Institute, Massachusetts, USA) and DeepVariant (Google, California, USA). Results Analysis of WGS data from the Platinum Genomes family revealed a total of 2,338,158 informative SNPs distributed evenly between grandparental haplotypes, of which 85,053 (3.6%) would have been detectable using the karyomapping SNP array. Analysis of the karyomapping window around common genes requiring PGT-M demonstrated an average 32x increase in the number of targets available for analysis in a PGT-M cycle. Analysis of 15x coverage WGS data in a model of embryo biopsy demonstrated an average 279,851 informative SNPs detected using standard variant calling and an average 285,107 using AI-based variant calling. The median distance between informative SNPs was reduced from 26,782 nucleotides to 1,212 nucleotides when comparing WGS-based karyomapping to SNP array-based karyomapping. Conclusions This study provides preclinical validation for the application of the karyomapping algorithm to WGS data. The markedly increased number of informative SNPs will allow successful feasibility studies for all couples presenting for PGT-M. The decreased distance between informative SNPs detected in a model embryo biopsy will improve diagnostic accuracy, particularly in detecting small double crossover events and reducing the number of cycles in which inadequate informative markers have been detected in the embryo sample. Finally, the use of WGS for diagnostic purposes, especially for rare and undiagnosed genetic disease, will allow the creation of a high resolution digital karyomap for couples with an affected child and improve the transition to IVF and PGT-M.

Meta-analysis of QTL for Fusarium head blight resistance in Chinese wheat landraces

Epidemics of Fusarium head blight (FHB), incited by Fusarium graminearum Schwabe, in wheat cause significant reductions in grain yield and quality. Numerous quantitative trait loci (QTL) for FHB resistance have been reported from Chinese sources. However, the relationships among QTL from different landraces have not been characterized. We earlier mapped QTL for FHB resistance using low-density maps developed from five recombinant inbred line (RIL) populations involving Chinese landraces ‘Haiyanzhong’ (HYZ), ‘Wangshuibai’ (WSB), ‘Baishanyuehuang’ (BSYH), ‘Huangfangzhu’ (HFZ), and ‘Huangcandou’ (HCD) as FHB resistant parents. In this study, we used maps of single nucleotide polymorphisms (SNP) developed from the five populations and identified 31 QTL on 16 chromosomes; 10 QTL were new. We constructed a consensus map and identified six meta-QTL (MQTL) and SNP within the MQTL regions using meta-analysis. Two of the MQTL were on chromosome 3BS (3BSd and 3BSc), and one on each of chromosomes 3A, 2D, 3D, and 4D. Twenty-two SNP closely linked to MQTL were converted into breeder friendly Kompetitive Allele Specific Polymerase Chain Reaction (KASP) assays, which should be useful for marker-assisted selection in breeding programs.

A study of mechanistic mapping of novel SNPs to male breast cancer.

Alterations in BRCA2, PALB2, CHEK2, and p53 genes have been identified for their association with male breast cancer in various studies. The incidence of male breast cancer in India is consistent with its global rate. The present study was carried out with an aim to evaluate the genetic alterations in male breast cancer patients from Malwa region of Punjab, India. Four male breast cancer patients belonging to different families were recruited from Guru Gobind Singh Medical College and Hospital, Faridkot, India. A total of 51 genes reported with implications in the pathogenesis of breast cancer were screened using next generation sequencing. Germline variations were found in BRCA1, BRCA2, PMS2, p53, and PALB2 genes, previously reported to be associated with MBC as well as FBC. In addition to these, 13 novel missense alterations were detected in eight genes including STK11, FZR1, PALB2, BRCA2, NF2, BAP1, BARD1, and CHEK2. Impact of these missense alterations on structure and function of protein was also analyzed through molecular dynamics simulation. Structural analysis of these single nucleotide polymorphisms (SNPs) revealed significant impact on the encoded protein functioning.

Quantitative trait locus mapping for plant height and branch number in an upland cotton recombinant inbred line with an SNP-based high-density genetic map

Cotton is one of the most important cash crops around the world, providing natural fiber for the textile industry. Agronomic traits play an important role in the mechanized harvesting of cotton. Plant height (PH) and branch number (BN) are two important traits that could affect plant architecture and ultimately, economic yield of cotton. The quantitative trait locus (QTL) for PH and BN across seven environments was identified with a high-density single nucleotide polymorphism map constructed using recombinant inbred lines from upland cotton 0–153 and sGK9708. A total of 68 QTLs for PH (nine stable) and 64 QTLs for BN (eight stable) were identified. Among these stable QTLs, three (two for PH and one for BN) have been identified in previous studies. Four hundred genes for PH and 624 genes for BN were located on the confidence intervals of these stable QTLs. Among them, 134 for PH and 224 for BN were expressed in at least one tissue of root, stem and leaf. Based on the annotation information, expression pattern and the function validated on the other species, ten genes (six for PH and four for BN) could be considered as potential candidate genes. These results could contribute to understanding the mechanism of PH and branch formation, and to improving the method of cotton breeding for molecular marker-assisted selection.

The first genetic linkage map for Fraxinus pennsylvanica and syntenic relationships with four related species.

The genetic linkage map for green ash (Fraxinus pennsylvanica) contains 1201 DNA markers in 23 linkage groups spanning 2008.87cM. The green ash map shows stronger synteny with coffee than tomato. Green ash (Fraxinus pennsylvanica) is an outcrossing, diploid (2n = 46) hardwood tree species, native to North America. Native ash species in North America are being threatened by the rapid spread of the emerald ash borer (EAB, Agrilus planipennis), an invasive pest from Asia. Green ash, the most widely distributed ash species, is severely affected by EAB infestation, yet few genomic resources for genetic studies and improvement of green ash are available. In this study, a total of 5712 high quality single nucleotide polymorphisms (SNPs) were discovered using a minimum allele frequency of 1% across the entire genome through genotyping-by-sequencing. We also screened hundreds of genomic- and EST-based microsatellite markers (SSRs) from previous de novo assemblies (Staton et al., PLoS ONE 10:e0145031, 2015; Lane et al., BMC Genom 17:702, 2016). A first genetic linkage map of green ash was constructed from 90 individuals in a full-sib family, combining 2719 SNP and 84 SSR segregating markers among the parental maps. The consensus SNP and SSR map contains a total of 1201 markers in 23 linkage groups spanning 2008.87cM, at an average inter-marker distance of 1.67cM with a minimum logarithm of odds of 6 and maximum recombination fraction of 0.40. Comparisons of the organization the green ash map with the genomes of asterid species coffee and tomato, and genomes of the rosid species poplar and peach, showed areas of conserved gene order, with overall synteny strongest with coffee.