Size QTL Research Articles

Genetic analysis and fine mapping of a grain size QTL in the small-grain sterile rice line Zhuo201S

The development and application of the small-grain rice sterile line Zhuo201S (Z201S) has demonstrated its potential for mechanized hybrid rice seed production, leading to significant cost reductions. However, the molecular mechanism responsible for the small-grain size characteristic of Z201S remains unclear. In this study, we conducted a genetic analysis using near-isogenic lines constructed from Z210S, a small-grain rice sterile line, and R2115, a normal-grain variety. The results revealed that the small-grain trait in Z201S is governed by a single partially dominant gene which also enhances grain number. Through mapping, we localized the causal gene to the short arm of chromosome 2, within a 113 kb physical region delimited by the molecular markers S2-4-1 and LB63. Transgenic analysis and gene expression assays indicated LOC_Os02g14760 as the most likely candidate gene, suggesting that the small-grain size trait of Z201S is controlled by a novel locus that has not been previously identified.

41 A preliminary investigation of candidate genes from a GWAS identified QTL for relative body size in the American Quarter Horse

Artificial selection during domestication has influenced an array of morphological traits in the horse, many that are now important economic traits for the horse industry. Growth and skeletal phenotypes impact the desirability and function of horses in diverse disciplines and are often key characteristics in the development of breeds. Skeletal traits also impact functionality: benefitting horses for specialized work or predisposing them to musculoskeletal injury. To investigate variations in body size and shape, we utilized measurements taken at 2 years of age (+/− one week) from 91 American Quarter Horses bred and owned by the University of Florida. We summarized 32 body measures from each horse using a principal component analysis followed by a varimax rotation. Factor 1 summarizes 19.5% of the variation and describes overall body size, except for head measurements which are better captured in Factor 2. Factor 1 traits encompass body proportions, heavily loading on the measure's height at dock and croup. DNA was extracted from blood and hair through previously published methods and genotyped at GeneSeek Inc. (Lansing, MI) using the Affymetrix Axiom Equine HD 670k array. We recently reported results from a preliminary genome-wide association study with 311,085 high-quality SNPs (poly-high resolution, MAF >0.05)suggesting a quantitative trait locus for the Body Factor 1 trait on chromosome 6 (raw P = 1.356e-09, Bonferroni P = 4.51e-07) and defines a candidate region spanning 1.5Mb. The QTL is in a region unrelated to a previously characterized HMGA2 body height locus. The candidate region contains 5 protein-coding genes, influencing human height and skeletal traits including one associated with Noonan Syndrome- a disease impacting height and facial morphology, an interesting parallel to the phenotype observed in our population. None of the 16,860 variants found in the European Nucleotide Archive for our candidate region had a functional impacted on a protein coding region. Therefore, we sequenced 2 whole genomes from American Quarter Horses to discover variants specific to the haplotype bearing this new body size QTL. Using the 10x Chromium library prep with 150bp paired end reads on an Illumina HiSeq2500, the 2 genomes totaled 1.5 billion reads. Following alignment and variant calling using the LongRanger package (v2.2, 10x Genomics Inc.), we observed 4,703 SNPs in the candidate region. Future investigation aims to investigate additional loci relevant to body morphological variations and develop predictive phenotype models in the horse.

Contribution of the grain size QTL GS3 to yield properties and physiological nitrogen-use efficiency in the large-grain rice cultivar 'Akita 63'.

The development of crop varieties with high nitrogen-use efficiency (NUE) is thought to be important in achieving sustainable cereal crop production. The high yield large-grain rice cultivar Oryza sativa L. ‘Akita 63’ (temperate japonica) has high physiological NUE (PNUE) for grain yield (GY). Our previous study revealed that a large-grain allele of GS3 is present in ‘Akita 63’. Here, we verified the influence of GS3 on the yield properties and PNUE for GY in ‘Akita 63’. The frequency distribution of brown rice length in F2 crosses of ‘Iwate 75’ and ‘Akita 63’ showed a continuous distribution that could be explained by GS3. A near-isogenic line was developed to substitute the GS3 segment of ‘Koshihikari’, which harbours a normal-sized grain allele, in the genetic background of ‘Akita 63’ and the line was designated as Akita63NILGS3-Koshihikari. Compared with Akita63NILGS3-Koshihikari, ‘Akita 63’ exhibited a significantly increased grain length, single brown grain weight and GY, although no significant differences were observed in the nitrogen content and above-ground biomass per unit of cultivated area. These results indicate that the GS3 large-grain allele is a contributing factor to high PNUE for GY in ‘Akita 63’. These findings will facilitate the development of nitrogen-efficient rice varieties.

34 Dimensionality of Genomic Information and Its Impact on GWA and Variant Selection: A Simulation Study

Abstract The ability to identify true-positive variants increases as more genotyped animals are available. Although thousands of animals can be genotyped, the dimensionality of the genomic information is limited. Therefore, there is a certain number of animals that represent all chromosome segments (Me) segregating in the population. The number of Me can be approximated from the eigenvalue decomposition of the genomic relationship matrix (G). Thus, the limited dimensionality may help to identify the number of animals to be used in genome-wide association (GWA). The first objective of this study was to examine different discovery set sizes for GWA, with set sizes based on the number of largest eigenvalues explaining a certain proportion of variance in G. Additionally, we investigated the impact of incorporating variants selected from different set sizes to regular SNP chip used for genomic prediction. Sequence data were simulated that contained 500k SNP and 2k QTL, where the genetic variance was fully explained by QTL. The GWA was conducted using the number of genotyped animals equal to the number of largest eigenvalues of G (EIG) explaining 50, 60, 70, 80, 90, 95, 98, and 99 percent of the variance in G. Significant SNP had a p-value lower than 0.05 with Bonferroni correction. Further, SNP with the largest effect size (top10, 100, 500, 1k, 2k, and 4k) were also selected to be incorporated into the 50k regular chip. Genomic predictions using the 50k combined with selected SNP were conducted using single-step GBLUP (ssGBLUP). Using the number of animals corresponding to at least EIG98 enabled the identification of the largest effect size QTL. The greatest accuracy of prediction was obtained when the top 2k SNP was combined to the 50k chip. The dimensionality of genomic information should be taken into account for variant selection in GWAS.

Fine-Mapping of a Wild Genomic Region Involved in Pod and Seed Size Reduction on Chromosome A07 in Peanut (Arachis hypogaea L.).

Fruit and seed size are important yield component traits that have been selected during crop domestication. In previous studies, Advanced Backcross Quantitative Trait Loci (AB-QTL) and Chromosome Segment Substitution Line (CSSL) populations were developed in peanut by crossing the cultivated variety Fleur11 and a synthetic wild allotetraploid (Arachis ipaensis × Arachis duranensis)4x. In the AB-QTL population, a major QTL for pod and seed size was detected in a ~5 Mb interval in the proximal region of chromosome A07. In the CSSL population, the line 12CS_091, which carries the QTL region and that produces smaller pods and seeds than Fleur11, was identified. In this study, we used a two-step strategy to fine-map the seed size QTL region on chromosome A07. We developed new SSR and SNP markers, as well as near-isogenic lines (NILs) in the target QTL region. We first located the QTL in ~1 Mb region between two SSR markers, thanks to the genotyping of a large F2 population of 2172 individuals and a single marker analysis approach. We then used nine new SNP markers evenly distributed in the refined QTL region to genotype 490 F3 plants derived from 88 F2, and we selected 10 NILs. The phenotyping of the NILs and marker/trait association allowed us to narrowing down the QTL region to a 168.37 kb chromosome segment, between the SNPs Aradu_A07_1148327 and Aradu_A07_1316694. This region contains 22 predicted genes. Among these genes, Aradu.DN3DB and Aradu.RLZ61, which encode a transcriptional regulator STERILE APETALA-like (SAP) and an F-box SNEEZY (SNE), respectively, were of particular interest. The function of these genes in regulating the variation of fruit and seed size is discussed. This study will contribute to a better knowledge of genes that have been targeted during peanut domestication.

Major seed size QTL on chromosome A05 of peanut (Arachis hypogaea) is conserved in the US mini core germplasm collection

Pod and seed size are important characteristics for the peanut industry and have been under strong selection pressure since peanut domestication. In order to dissect the genetic control of peanut pod and seed size, a recombinant inbred mapping population from a cross of Florida-07 by GP-NC WS 16 was used to determine the genomic regions associated with traits including 100 pod weight, 100 seed weight, pod weight of double-seeded pods, seed weight of double-seeded pods, and area of double-seeded pods. Nine QTL on linkage groups (LGs) A05, A06, A09, B10, B04, A03, B05, and B08 were associated with pod and seed size. A majority of the QTL have small effects except the locus on LG A05 (93 to 102 Mbp) which explained up to 66% phenotypic variation for all measured pod and seed traits. A comparison of QTL previously reported for yield component traits showed a common QTL on LG A05 was detected in two genetic populations whose parentage is distinct from those used in this study. The markers tightly linked to this major QTL were informative in distinguishing large versus small-seeded germplasm lines in the mini core collection originating from thirty-one countries, suggesting selection for this seed size QTL in large-seeded ecotypes. However, the large seed size allele appeared to co-segregate with a late leaf spot disease susceptibility allele inherited from the Florida-07 parent. Therefore, peanut breeders need to weigh the pros and cons before integrating the large seed size QTL from Florida-07 in their breeding program.

Large-scale GWAS in sorghum reveals common genetic control of grain size among cereals.

SummaryGrain size is a key yield component of cereal crops and a major quality attribute. It is determined by a genotype’s genetic potential and its capacity to fill the grains. This study aims to dissect the genetic architecture of grain size in sorghum. An integrated genome‐wide association study (GWAS) was conducted using a diversity panel (n = 837) and a BC‐NAM population (n = 1421). To isolate genetic effects associated with genetic potential of grain size, rather than the genotype’s capacity to fill the grains, a treatment of removing half of the panicle was imposed during flowering. Extensive and highly heritable variation in grain size was observed in both populations in 5 field trials, and 81 grain size QTL were identified in subsequent GWAS. These QTL were enriched for orthologues of known grain size genes in rice and maize, and had significant overlap with SNPs associated with grain size in rice and maize, supporting common genetic control of this trait among cereals. Grain size genes with opposite effect on grain number were less likely to overlap with the grain size QTL from this study, indicating the treatment facilitated identification of genetic regions related to the genetic potential of grain size. These results enhance understanding of the genetic architecture of grain size in cereal, and pave the way for exploration of underlying molecular mechanisms and manipulation of this trait in breeding practices.

Analysis of porcine body size variation using re-sequencing data of miniature and large pigs

BackgroundDomestication has led to substantial phenotypic and genetic variation in domestic animals. In pigs, the size of so called minipigs differs by one order of magnitude compared to breeds of large body size. We used biallelic SNPs identified from re-sequencing data to compare various publicly available wild and domestic populations against two minipig breeds to gain better understanding of the genetic background of the extensive body size variation. We combined two complementary measures, expected heterozygosity and the composite likelihood ratio test implemented in “SweepFinder”, to identify signatures of selection in Minipigs. We intersected these sweep regions with a measure of differentiation, namely FST, to remove regions of low variation across pigs. An extraordinary large sweep between 52 and 61 Mb on chromosome X was separately analyzed based on SNP-array data of F2 individuals from a cross of Goettingen Minipigs and large pigs.ResultsSelective sweep analysis identified putative sweep regions for growth and subsequent gene annotation provided a comprehensive set of putative candidate genes. A long swept haplotype on chromosome X, descending from the Goettingen Minipig founders was associated with a reduction of adult body length by 3% in F2 cross-breds.ConclusionThe resulting set of genes in putative sweep regions implies that the genetic background of body size variation in pigs is polygenic rather than mono- or oligogenic. Identified genes suggest alterations in metabolic functions and a possible insulin resistance to contribute to miniaturization. A size QTL located within the sweep on chromosome X, with an estimated effect of 3% on body length, is comparable to the largest known in pigs or other species. The androgen receptor AR, previously known to influence pig performance and carcass traits, is the most obvious potential candidate gene within this region.

「Rialto」由来の多収・大粒QTLを導入した秋まき小麦系統の農業特性

「Rialto」由来の多収・大粒QTLを導入した秋まき小麦系統の農業特性

QTL mapping of domestication and diversifying selection related traits in round-fruited semi-wild Xishuangbanna cucumber (Cucumis sativus L. var. xishuangbannanesis)

QTL analysis revealed 11 QTL underlying flowering time and fruit size variation in the semi-wild Xishuangbanna cucumber, of which, FT6.2 and FS5.2 played the most important roles in determining photoperiod-dependent flowering time and round-fruit shape, respectively. Flowering time and fruit size are two important traits in domestication and diversifying selection in cucumber, but their genetic basis is not well understood. Here we reported QTL mapping results on flowering time and fruit size with F2 and F2:3 segregating populations derived from the cross between WI7200, a small fruited, early flowering primitive cultivated cucumber and WI7167, a round-fruited, later flowering semi-wild Xishuangbanna (XIS) cucumber. A linkage map with 267 microsatellite marker loci was developed with 138 F2 plants. Phenotypic data of male and female flowering time, fruit length and diameter and three other traits (mature fruit weight and number, and seedling hypocotyl length) were collected in multiple environments. Three flowering time QTL, FT1.1, FT5.1 and FT6.2 were identified, in which FT6.2 played the most important role in conferring less photoperiod sensitive early flowering during domestication whereas FT1.1 seemed more influential in regulating flowering time within the cultivated cucumber. Eight consensus fruit size QTL distributed in 7 chromosomes were detected, each of which contributed to both longitudinal and radial growth in cucumber fruit development. Among them, FS5.2 on chromosome 5 exhibited the largest effect on the determination of round fruit shape that was characteristic of the WI7167 XIS cucumber. Possible roles of these flowering time and fruit size QTL in domestication of cucumber and crop evolution of the semi-wild XIS cucumber, as well as the genetic basis of round fruit shape in cucumber are discussed.

Reciprocal translocation identified in Vigna angularis dominates the wild population in East Japan.

Using an F2 population derived from cultivated and wild azuki bean, we previously detected a reciprocal translocation and a seed size QTL near the translocation site. To test the hypothesis that the translocation in the cultivated variety contributed to the larger seed size, we performed further linkage analyses with several cross combinations between cultivated and wild azuki beans. In addition, we visually confirmed the translocation by cytogenetic approach using 25 wild and cultivated accessions. As a result, we found the translocation-type chromosomes in none of the cultivated accessions, but in a number of the wild accessions. Interestingly, all the wild accessions with the translocation were originally collected from East Japan, while all the accessions with normal chromosomes were from West Japan or the Sea of Japan-side region. Such biased geographical distribution could be explained by the glacial refugium hypothesis, and supported narrowing down the domestication origin of cultivated azuki bean.

Validation of linked QTL for bacterial cold water disease resistance and spleen size on rainbow trout chromosome Omy19

Abstract Bacterial cold water disease (BCWD) is caused by infection with Flavobacterium psychrophilum , and outbreaks cause significant economic losses in salmonid aquaculture. Previously, we identified a major QTL for BCWD resistance on rainbow trout chromosome Omy19 as well as a QTL for spleen size (SPLW = spleen weight and SPLI = spleen index) which in naive fish is a surrogate indicator of BCWD resistance. The objective of this study was to validate the BCWD resistance and spleen size QTL in a subsequent generation and investigate whether the correlation between BCWD resistance and spleen size loci on Omy19 was due to close linkage or pleiotropy. We designed four full-sib matings whose offspring were evaluated for BCWD resistance, body weight, SPLW and SPLI. A total of 800 offspring were genotyped with seven microsatellite markers from Omy19. Our analyses confirmed the presence of QTL for BCWD and spleen size on Omy19. Our findings also suggest that BCWD resistance and spleen size are not influenced by a single pleiotropic QTL on Omy19 as the QTL were separately inherited in the families studied. These data increase our understanding of the genetic basis of rainbow trout BCWD resistance.

QTL modulating ear size and erectness in pigs

Ear size and erectness are important conformation measurements in pigs. An F(2) population established by crossing European Large White (small, erect ears) with Chinese Meishan (large, flop ears) was used to study the genetic influence of the two ear traits for the first time. A linkage map incorporating 152 markers on 18 autosomal chromosomes was utilised in a genome scan for QTL. Significant QTL were found on SSC1, 5, 7, 9 and 12 for the two traits. The QTL on SSC5 and SSC7 had major effects and were significant at the genome-wide level (P < 0.01). The QTL on SSC1 for ear erectness also had a major effect and was genome-wide significant (P < 0.01). The 95% confidence interval (CI) of the ear size QTL on SSC5 spanned only 4 cM. The QTL on SSC7 for the two ear traits each had a CI of <20 cM, and their positions overlapped with those of the major QTL affecting subcutaneous fat depths on the same chromosome. This study provides insights on the complex genetic influences underlying pig ear traits and will facilitate positional candidate gene analysis to identify causative DNA variants.

Chromosomal anchoring of linkage groups and identification of wing size QTL using markers and FISH probes derived from microdissected chromosomes in Nasonia (Pteromalidae: Hymenoptera)

Nasonia vitripennis is a small parasitic hymenopteran with a 50-year history of genetic work including linkage mapping with mutant and molecular markers. For the first time we are now able to anchor linkage groups to specific chromosomes. Two linkage maps based on a hybrid cross (N. vitripennis × N. longicornis) were constructed using STS, RAPD and microsatellite markers, where 17 of the linked STS markers were developed from single microdissected banded chromosomes. Based on these microdissections we anchored all linkage groups to the five chromosomes of N. vitripennis. We also verified the chromosomal specificity of the microdissection through in situ hybridization and linkage analyses. This information and technique will allow us in the future to locate genes or QTL detected in different mapping populations efficiently and fast on homologous chromosomes or even chromosomal regions. To test this approach we asked whether QTL responsible for the wing size in two different hybrid crosses (N. vitripennis × N. longicornis and N. vitripennis × N.giraulti) map to the same location. One QTL with a major effect was found to map to the centromere region of chromosome 3 in both crosses. This could indicate that indeed the same gene/s is involved in the reduction of wing in N. vitripennis and N. longicornis.

Identification of quantitative trait loci for prolificacy and growth in mice.

Marker-quantitative trait locus (QTL) linkage was evaluated in F2 intercross and backcross mouse populations derived from stocks differing dramatically in prolificacy and mature weight. A highly prolific outbred Quackenbush-Swiss mouse line, or an inbred line derived from it (16.62 +/- 0.22 and 14.64 +/- 0.27 pups per litter, respectively) were used as one of the grandparents in these populations. The less prolific C57BL/6J inbred mouse line (6.67 +/- 0.37 pups per litter) was used as the other grandparent. Linkage was evaluated in a three-step process that involved selective genotyping of F2 intercross progeny representing extremes for prolificacy, genotyping of the full F2 for chromosomal regions potentially associated with prolificacy, and genotyping of the backcross for genomic regions significantly associated with prolificacy in the F2. Segments of Chromosomes (Chrs) 2 and 4 were significantly (P < 0.05, experiment-wise error rate) associated with prolificacy, and LOD scores suggestive of linkage were observed for litter size on Chr 9 and growth on Chrs 4 and 11. Existence of growth QTL was also supported by marker effects that were significant (P < 0.05) or approaching significance (P < 0.10) in the backcross. Additive litter size QTL effects ranged from 0.56 to 0.79 pups per litter, and dominance deviations ranged from -0.56 to 1.19 pups per litter, suggesting overdominance as a possible mode of gene action in some cases. The observation of pleiotropic or linked QTL for growth and prolificacy corresponds well with results from many selection experiments identifying positively correlated responses to selection for these two traits.