Empirical threshold values for quantitative trait mapping.

Genome-wide association studies (GWAS) based on linkage disequilibrium (LD) provide a promising tool for the detection and fine mapping of quantitative trait loci (QTL) underlying complex agronomic traits. In this study we explored the genetic basis of variation for the traits heading date, plant height, thousand grain weight, starch content and crude protein content in a diverse collection of 224 spring barleys of worldwide origin. The whole panel was genotyped with a customized oligonucleotide pool assay containing 1536 SNPs using Illumina's GoldenGate technology resulting in 957 successful SNPs covering all chromosomes. The morphological trait "row type" (two-rowed spike vs. six-rowed spike) was used to confirm the high level of selectivity and sensitivity of the approach. This study describes the detection of QTL for the above mentioned agronomic traits by GWAS. Population structure in the panel was investigated by various methods and six subgroups that are mainly based on their spike morphology and region of origin. We explored the patterns of linkage disequilibrium (LD) among the whole panel for all seven barley chromosomes. Average LD was observed to decay below a critical level (r2-value 0.2) within a map distance of 5-10 cM. Phenotypic variation within the panel was reasonably large for all the traits. The heritabilities calculated for each trait over multi-environment experiments ranged between 0.90-0.95. Different statistical models were tested to control spurious LD caused by population structure and to calculate the P-value of marker-trait associations. Using a mixed linear model with kinship for controlling spurious LD effects, we found a total of 171 significant marker trait associations, which delineate into 107 QTL regions. Across all traits these can be grouped into 57 novel QTL and 50 QTL that are congruent with previously mapped QTL positions. Our results demonstrate that the described diverse barley panel can be efficiently used for GWAS of various quantitative traits, provided that population structure is appropriately taken into account. The observed significant marker trait associations provide a refined insight into the genetic architecture of important agronomic traits in barley. However, individual QTL account only for a small portion of phenotypic variation, which may be due to insufficient marker coverage and/or the elimination of rare alleles prior to analysis. The fact that the combined SNP effects fall short of explaining the complete phenotypic variance may support the hypothesis that the expression of a quantitative trait is caused by a large number of very small effects that escape detection. Notwithstanding these limitations, the integration of GWAS with biparental linkage mapping and an ever increasing body of genomic sequence information will facilitate the systematic isolation of agronomically important genes and subsequent analysis of their allelic diversity.

/pdf/genome-wide-association-studies-for-agronomical-traits-in-a-3vwfogjrxx.pdf

Genome-wide association studies for agronomical traits in a world wide spring barley collection.

The first genetic transcript map of the soybean genome was created by mapping one SNP in each of 1141 genes in one or more of three recombinant inbred line mapping populations, thus providing a picture of the distribution of genic sequences across the mapped portion of the genome Single-nucleotide polymorphisms (SNPs) were discovered via the resequencing of sequence-tagged sites (STSs) developed from expressed sequence tag (EST) sequence From an initial set of 9459 polymerase chain reaction primer sets designed to a diverse set of genes, 4240 STSs were amplified and sequenced in each of six diverse soybean genotypes In the resulting 244 Mbp of aligned sequence, a total of 5551 SNPs were discovered, including 4712 single-base changes and 839 indels for an average nucleotide diversity of θ = 0000997 The analysis of the observed genetic distances between adjacent genes vs the theoretical distribution based upon the assumption of a random distribution of genes across the 20 soybean linkage groups clearly indicated that genes were clustered Of the 1141 genes, 291 mapped to 72 of the 112 gaps of 5–10 cM in the preexisting simple sequence repeat (SSR)-based map, while 111 genes mapped in 19 of the 26 gaps >10 cM The addition of 1141 sequence-based genic markers to the soybean genome map will provide an important resource to soybean geneticists for quantitative trait locus discovery and map-based cloning, as well as to soybean breeders who increasingly depend upon marker-assisted selection in cultivar improvement

/pdf/a-soybean-transcript-map-gene-distribution-haplotype-and-5fchvmharh.pdf

A soybean transcript map: gene distribution, haplotype and single-nucleotide polymorphism analysis.

Summary
We have studied the nucleotide diversity of common bean, Phaseolus vulgaris, which is characterized by two independent domestications in two geographically distinct areas: Mesoamerica and the Andes. This provides an important model, as domestication can be studied as a replicate experiment.
We used nucleotide data from five gene fragments characterized by large introns to analyse 214 accessions (102 wild and 112 domesticated). The wild accessions represent a cross-section of the entire geographical distribution of P. vulgaris.
A reduction in genetic diversity in both of these gene pools was found, which was three-fold greater in Mesoamerica compared with the Andes. This appears to be a result of a bottleneck that occurred before domestication in the Andes, which strongly impoverished this wild germplasm, leading to the minor effect of the subsequent domestication bottleneck (i.e. sequential bottleneck).
These findings show the importance of considering the evolutionary history of crop species as a major factor that influences their current level and structure of genetic diversity. Furthermore, these data highlight a single domestication event within each gene pool. Although the findings should be interpreted with caution, this evidence indicates the Oaxaca valley in Mesoamerica, and southern Bolivia and northern Argentina in South America, as the origins of common bean domestication.

https://nph.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1469-8137.2012.04377.x

Molecular analysis of the parallel domestication of the common bean (Phaseolus vulgaris) in Mesoamerica and the Andes.

Sucrose synthase catalyzes the reaction sucrose + UDP → UDP-glucose + fructose, the first step in the conversion of sucrose to starch in endosperm. Previous studies identified two tissue-specific, yet functionally redundant, sucrose synthase (SUS) genes, Sus1 and Sus2. In the present study, the wheat Sus2 orthologous gene (TaSus2) series was isolated and mapped on chromosomes 2A, 2B, and 2D. Based on sequencing in 61 wheat accessions, three single-nucleotide polymorphisms (SNPs) were detected in TaSus2-2B. These formed two haplotypes (Hap-H and Hap-L), but no diversity was found in either TaSus2-2A or TaSus2-2D. Based on the sequences of the two haplotypes, we developed a co-dominant marker, TaSus2-2B tgw , which amplified 423 or 381-bp fragments in different wheat accessions. TaSus2-2B tgw was located between markers Xbarc102.2 and Xbarc91 on chromosome 2BS in a RIL population from Xiaoyan 54 × Jing 411. Association analysis suggested that the two haplotypes were significantly associated with 1,000 grain weight (TGW) in 89 modern wheat varieties in the Chinese mini-core collection. Mean TGW difference between the two haplotypes over three cropping seasons was 4.26 g (varying from 3.71 to 4.94 g). Comparative genomics analysis detected major kernel weight QTLs not only in the chromosome region containing TaSus2-2B tgw, but also in the collinear regions of TaSus2 on rice chromosome 7 and maize chromosome 9. The preferred Hap-H haplotype for high TGW underwent very strong positive selection in Chinese wheat breeding, but not in Europe. The geographic distribution of Hap-H was perhaps determined by both latitude and the intensity of selection in wheat breeding.

The wheat (T. aestivum) sucrose synthase 2 gene (TaSus2) active in endosperm development is associated with yield traits.

The least ambiguous genetic markers are those based on completely characterized DNA sequence polymorphisms. Unfortunately, assaying allele states by allele sequencing is slow and cumbersome. The most desirable type of genetic marker would be unambiguous, inexpensive to assay and would be assayable singly or in parallel with hundreds of other markers (multiplexable). In this report we sequenced alleles at 54 barley (Hordeum vulgare ssp. vulgare) loci, 38 of which contained single-nucleotide polymorphisms (SNPs). Many of these 38 loci contained multiple polymorphisms, and a total of 112 polymorphisms were scored in five barley genotypes. The polymorphism data set was analyzed both by using the individual mutations as cladistic characters and by reducing data for each locus to haplotypes. We compared the informativeness of these two approaches by consensus tree construction and bootstrap analysis. Both approaches provided similar results. Since some of the loci sequenced contained insertion/deletion events and multiple point mutations, we thought that these multiple-mutated loci might represent old alleles that predated the divergence of barley from H. spontaneum. We evaluated sequences from a sample of H. spontaneum accessions from the Eastern Mediterranean, and observed similar alleles present in both cultivated barley and H. spontaneum, suggesting either multiple domestication events or multiple transfers of genes between barley and its wild ancestor.

Discovery and assay of single-nucleotide polymorphisms in barley (Hordeum vulgare).

Single-nucleotide polymorphisms (SNPs) represent the most prevalent class of genetic markers available for linkage disequilibrium or cladistic analyses. PCR primers may be labeled with fluorescent dyes and used to rapidly and accurately differentiate among alleles that are defined by a single-nucleotide differences. Here, we describe the primer-mediated detection of SNPs based on primer mismatch during allele-specific amplification of preamplified target sequences. Primers are labeled with different fluors at their 5' nucleotides, with their 3' termini at the transition mutation that defines allelic variation at the target locus. Each primer perfectly matches one of the two available alleles for each locus. Electrophoretic detection permits characterization of the product both by size and fluor. This report demonstrates some of the capabilities of this assay, including heterozygote determination and multiplexed analysis.

https://www.future-science.com/doi/pdf/10.2144/00284st07

Electrophoretic detection of single-nucleotide polymorphisms

Cultivated barley is the major livestock feed grain in the Northern Plains and Northwestern United States due to the fact that its short growing season and limited rainfall limit the planting and production of corn. Starch and fiber content play a significant role in feedlot performance of animals raised on barley feed. To study the underlying genetic locations and mechanisms for these traits, a recombinant inbred line population was derived from a cross between the hulled barley cultivar Valier and a hull-less Swiss landrace line, PI370970. Valier has a high acid detergent fiber content (ADF) and low starch and protein while PI370970 contains low ADF and high starch and protein content. To detect associated QTLs, data were collected and analyzed from irrigated and rain-fed environments. A total of 30 main effect QTLs and four epistatic QTLs were identified which conditioned ADF, starch and protein content under rain-fed, irrigated and combined analyses. These QTLs were located on chromosomes 2H, 3H, 5H, 6H and 7H. Major ADF and starch QTL were identified on chromosome 7H near the nud locus (the locus controlling hulled vs. hull-less caryopsis). High heritability estimates for both ADF and starch content suggest that early selection for these traits during breeding would be productive. Low ADF-QTL were independently verified in a second population in a different genetic background.

Quantitative trait loci of acid detergent fiber and grain chemical composition in hulled × hull-less barley population

More than half of the barley grown in the USA is used for livestock feed, with the remaining stocks diverted for human food and malting purposes. The use of barley grain as a major source of cattle feed has been criticized because of its rapid digestion in the rumen, which can result in digestive disorders in cattle. In sacco dry matter digestibility (ISDMD) and particle size (PS) after dry rolling have been found to play a role in the feedlot performance of barley as a feed grain. Reducing the rate of ISDMD is predicted to result in significantly improved animal health and average daily gain. A recombinant inbred line population derived from a cross between a high ISDMD, two-rowed barley cultivar (Valier) and a six-rowed Swiss landrace line (PI370970) exhibiting far slower ISDMD has been developed for studying the underlying genetic locations and mechanisms of these traits. To detect associated quantitative trait loci (QTLs), we collected and analyzed data from irrigated and rain-fed environments. A significant negative correlation was observed between ISDMD and PS. High heritability estimates for ISDMD and PS suggest that early selection for these traits during breeding would be achievable. Four QTLs were identified on chromosomes 2H, 6H, and 7H, explaining 73–85% of ISDMD phenotypic variation, while three QTLs on 2H and 7H were associated with variation in PS and explained 58–77% of its variation. A major QTL on chromosome 2H tightly linked to the morphology-modifying gene vrs1 was found to dramatically control 35–62% of the phenotypic variation of ISDMD and 26–53% of that of PS. The impact of the vrs1 locus on ISDMD was validated in two populations representing different genetic backgrounds. Our results suggest that it may also be advantageous to simultaneously overlap these QTLs around the vrs1 locus.

Quantitative trait loci for dry matter digestibility and particle size traits in two-rowed × six-rowed barley population

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Papers

Discovery and assay of single-nucleotide polymorphisms in barley (Hordeum vulgare).

Electrophoretic detection of single-nucleotide polymorphisms

Quantitative trait loci of acid detergent fiber and grain chemical composition in hulled × hull-less barley population

Quantitative trait loci for dry matter digestibility and particle size traits in two-rowed × six-rowed barley population