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      Fine mapping and discovery of candidate genes for seed size in watermelon by genome survey sequencing

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      Scientific Reports
      Nature Publishing Group UK

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          Abstract

          Fine mapping and discovery of candidate genes underlying seed size are important for modern watermelon breeding. Here, by using a high-resolution genetic map and whole-genome genetic variation detection aided by genome survey sequencing, we fine mapped and discovered candidate genes for seed size in watermelon. QTL (quantitative trait locus) mapping identified two pleiotropic QTLs for seed size, namely, qSS4 and qSS6, using a high-density genetic map constructed by specific length amplified fragment sequencing. qSS6 explained 93.00%, 94.11% and 95.26% of the phenotypic variation in thousand-seed weight, seed length and seed width, respectively, and was defined as a major QTL. Then, high-coverage re-sequencing of two parental lines detected a total of 193,395 SNPs (single nucleotide polymorphisms) and 45,065 indels (insertions/deletions), which corresponded to a frequency of 534 SNPs/Mb and 124 indels/Mb. Based on the genetic variation in the two parental lines, newly developed PCR-based markers allowed the region of qSS6 to be narrowed to 55.5 kb. Three potential candidates were identified, including a known seed size regulator in rice, SRS3. Taken together, our results reveal successful rapid fine mapping and discovery of candidate genes for seed size in watermelon, which could be applied to many traits of interest in plants.

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          Precision mapping of quantitative trait loci.

          Adequate separation of effects of possible multiple linked quantitative trait loci (QTLs) on mapping QTLs is the key to increasing the precision of QTL mapping. A new method of QTL mapping is proposed and analyzed in this paper by combining interval mapping with multiple regression. The basis of the proposed method is an interval test in which the test statistic on a marker interval is made to be unaffected by QTLs located outside a defined interval. This is achieved by fitting other genetic markers in the statistical model as a control when performing interval mapping. Compared with the current QTL mapping method (i.e., the interval mapping method which uses a pair or two pairs of markers for mapping QTLs), this method has several advantages. (1) By confining the test to one region at a time, it reduces a multiple dimensional search problem (for multiple QTLs) to a one dimensional search problem. (2) By conditioning linked markers in the test, the sensitivity of the test statistic to the position of individual QTLs is increased, and the precision of QTL mapping can be improved. (3) By selectively and simultaneously using other markers in the analysis, the efficiency of QTL mapping can be also improved. The behavior of the test statistic under the null hypothesis and appropriate critical value of the test statistic for an overall test in a genome are discussed and analyzed. A simulation study of QTL mapping is also presented which illustrates the utility, properties, advantages and disadvantages of the method.
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            A rapid DNA isolation procedure for small quantities of freshleaf tissue

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              The TRANSPARENT TESTA12 gene of Arabidopsis encodes a multidrug secondary transporter-like protein required for flavonoid sequestration in vacuoles of the seed coat endothelium.

              Phenolic compounds that are present in the testa interfere with the physiology of seed dormancy and germination. We isolated a recessive Arabidopsis mutant with pale brown seeds, transparent testa12 (tt12), from a reduced seed dormancy screen. Microscopic analysis of tt12 developing and mature testas revealed a strong reduction of proanthocyanidin deposition in vacuoles of endothelial cells. Double mutants with tt12 and other testa pigmentation mutants were constructed, and their phenotypes confirmed that tt12 was affected at the level of the flavonoid biosynthetic pathway. The TT12 gene was cloned and found to encode a protein with similarity to prokaryotic and eukaryotic secondary transporters with 12 transmembrane segments, belonging to the MATE (multidrug and toxic compound extrusion) family. TT12 is expressed specifically in ovules and developing seeds. In situ hybridization localized its transcript in the endothelium layer, as expected from the effect of the tt12 mutation on testa flavonoid pigmentation. The phenotype of the mutant and the nature of the gene suggest that TT12 may control the vacuolar sequestration of flavonoids in the seed coat endothelium.
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                Author and article information

                Contributors
                mashuangwu@caas.cn
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                14 December 2018
                14 December 2018
                2018
                : 8
                : 17843
                Affiliations
                GRID grid.464499.2, Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, The Laboratory of Melon Crops, ; Zhengzhou, Henan Province 450009 China
                Author information
                http://orcid.org/0000-0001-5311-2418
                Article
                36104
                10.1038/s41598-018-36104-w
                6294751
                30552379
                a4df84c7-8430-44a3-9414-bced597c8940
                © The Author(s) 2018

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 7 June 2018
                : 14 November 2018
                Funding
                Funded by: Technology Innovation Program of China (CAAS-ASTIP-2017-ZFRI)
                Funded by: the National Key R&D Program of China (2016YFD0100204-26), the Special Protection and Utilization of the Crop Germplasm Resources of China (2014NWB038), the National R&D Infrastructure and Facility Development Program of China and the Agricultural Science and Technology Innovation Program of China (CAAS-ASTIP-2017-ZFRI)
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