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      Mapping Quantitative Trait Loci onto Chromosome-Scale Pseudomolecules in Flax

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          Abstract

          Quantitative trait loci (QTL) are genomic regions associated with phenotype variation of quantitative traits. To date, a total of 313 QTL for 31 quantitative traits have been reported in 14 studies on flax. Of these, 200 QTL from 12 studies were identified based on genetic maps, the scaffold sequences, or the pre-released chromosome-scale pseudomolecules. Molecular markers for QTL identification differed across studies but the most used ones were simple sequence repeats (SSRs) or single nucleotide polymorphisms (SNPs). To uniquely map the SSR and SNP markers from different references onto the recently released chromosome-scale pseudomolecules, methods with several scripts and database files were developed to locate PCR- and SNP-based markers onto the same reference, co-locate QTL, and scan genome-wide candidate genes. Using these methods, 195 out of 200 QTL were successfully sorted onto the 15 flax chromosomes and grouped into 133 co-located QTL clusters; the candidate genes that co-located with these QTL clusters were also predicted. The methods and tools presented in this article facilitate marker re-mapping to a new reference, genome-wide QTL analysis, candidate gene scanning, and breeding applications in flax and other crops.

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          Methods for linkage disequilibrium mapping in crops.

          Linkage disequilibrium (LD) mapping in plants detects and locates quantitative trait loci (QTL) by the strength of the correlation between a trait and a marker. It offers greater precision in QTL location than family-based linkage analysis and should therefore lead to more efficient marker-assisted selection, facilitate gene discovery and help to meet the challenge of connecting sequence diversity with heritable phenotypic differences. Unlike family-based linkage analysis, LD mapping does not require family or pedigree information and can be applied to a range of experimental and non-experimental populations. However, care must be taken during analysis to control for the increased rate of false positive results arising from population structure and variety interrelationships. In this review, we discuss how suitable the recently developed alternative methods of LD mapping are for crops.
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            Sequence mapping by electronic PCR.

            The highly specific and sensitive PCR provides the basis for sequence-tagged sites (STSs), unique landmarks that have been used widely in the construction of genetic and physical maps of the human genome. Electronic PCR (e-PCR) refers to the process of recovering these unique sites in DNA sequences by searching for subsequences that closely match the PCR primers and have the correct order, orientation, and spacing that they could plausibly prime the amplification of a PCR product of the correct molecular weight. A software tool was developed to provide an efficient implementation of this search strategy and allow the sort of en masse searching that is required for modern genome analysis. Some sample searches were performed to demonstrate a number of factors that can affect the likelihood of obtaining a match. Analysis of one large sequence database record revealed the presence of several microsatellite and gene-based markers and allowed the exact base-pair distances among them to be calculated. This example provides a demonstration of how e-PCR can be used to integrate the growing body of genomic sequence data with existing maps, reveal relationships among markers that existed previously on different maps, and correlate genetic distances with physical distances.
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              Believe it or not, QTLs are accurate!

              It is generally believed that mapping quantitative trait loci (QTLs) does not accurately position genes underlying polygenic traits on the genome, which limits the application of QTL analysis in marker-assisted selection and gene discovery. However, now that a few plant QTLs have been cloned or accurately tagged, it appears that they might be accurate to within 2cM or less. This means that there will be circumstances when map-based cloning using only original mapping data would be a realistic option that avoids time-consuming and expensive fine mapping. Acceptance of this view would enhance the value of past and future mapping experiments, particularly those revealing small and environmentally sensitive QTLs that are often considered intractable at the molecular level.
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                Author and article information

                Journal
                Methods Protoc
                Methods Protoc
                mps
                Methods and Protocols
                MDPI
                2409-9279
                04 April 2020
                June 2020
                : 3
                : 2
                : 28
                Affiliations
                Ottawa Research and Development Centre, Agriculture and Agri-Food Canada, Ottawa, ON K1A 0C6, Canada; sylviej.cloutier@ 123456canada.ca
                Author notes
                [* ]Correspondence: frank.you@ 123456canada.ca ; Tel.: +1-613-759-1539
                Author information
                https://orcid.org/0000-0003-4424-6179
                Article
                mps-03-00028
                10.3390/mps3020028
                7359702
                32260372
                e196dbf8-a8cd-45ef-9443-4e345ba62cea
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 10 March 2020
                : 02 April 2020
                Categories
                Technical Note

                flax,association mapping,genome-wide association study (gwas),simple sequence repeat (ssr),single nucleotide polymorphism (snp),quantitative trait loci (qtl),chromosome-scale pseudomolecules

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