• Title/Summary/Keyword: Marker selection

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Development of AFLP and STS Markers Related to Stay Green Trait in Multi-Tillered Maize

  • Jang Cheol Seong;Lee Hee Bong;Seo Yong Weon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.358-362
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    • 2004
  • In order to develop molecular markers related to stay green phenotype, AFLP analysis was conducted using near-isogenic lines for either stay green or non stay green trait. Both lines have characteristics of multi-ear and tillers (MET). Two out of 64 primer combinations of selective amplification identified three reproducible polymorphic fragments in MET corn with stay green. Both of E+AGC/M+CAC and E+AAG/M+CAA primer combinations produced two and one specific polymorphic fragments linked to stay green trait, respectively. For the conversion of AFLPs to sequence tag sites (STSs), primers were designed form both end sequences of each two polymorphic fragments. One fragment, which was amplified with E+AAG/M+CAA primer combinations, possessed 298 bp long and showed a $91\%$ homology with maize retrotransposon Cinful-l. One out of two polymorphic fragments produced with E+AGC/M+CAC primer combination had 236 bp long and matched a $96\%$ homology with an intron region of 22kDa alpha zein gene cluster in Zea mays. One out of two PCR fragments amplified with MET2 primer set in the stay green MET was not produced in the non-stay green MET. The developed AFLP and STS marker could be used as an efficient tool for selection of the stay green trait in the MET inbred.

High-density genetic mapping using GBS in Chrysanthemum

  • Chung, Yong Suk;Cho, Jin Woong;Kim, Changsoo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.57-57
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    • 2017
  • Chrysanthemum is one of the most important floral crop in Korea produced about 7 billion dollars (1 billion for pot and 6 billion for cutting) in 2013. However, it is difficult to breed and to do genetic study because 1) it is highly self-incompatible, 2) it is outcrossing crop having heterozygotes, and 3) commercial cultvars are hexaploid (2n = 6x = 54). Although low-density genetic map and QTL study were reported, it is not enough to apply for the marker assisted selection and other genetic studies. Therefore, we are trying to make high-density genetic mapping using GBS with about 100 $F_1s$ of C. boreale that is oHohhfd diploid (2n = 2x = 18, about 2.8Gb) instead of commercial culitvars. Since Chrysanthemum is outcrossing, two-way pseudo-testcross model would be used to construct genetic map. Also, genotype-by-sequencing (GBS) would be utilized to generate sufficient number of markers and to maximize genomic representation in a cost effective manner. Those completed sequences would be analyzed with TASSEL-GBS pipeline. In order to reduce sequence error, only first 64 sequences, which have almost zero percent error, would be incorporated in the pipeline for the analysis. In addition, to reduce errors that is common in heterozygotes crops caused by low coverage, two rare cutters (NsiI and MseI) were used to increase sequence depth. Maskov algorithm would also used to deal with missing data. Further, sparsely placed markers on the physical map would be used as anchors to overcome problems caused by low coverage. For this purpose, were generated from transcriptome of Chrysanthemum using MISA program. Among those, 10 simple sequence repeat (SSR) markers, which are evenly distributed along each chromosome and polymorphic between two parents, would be selected.

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Identification and molecular characterization of downy mildew resistant gene candidates in maize (Zea mays subsp. Mays)

  • Kim, Jae Yoon;Kim, Chang-Ho;Kim, Kyung Hee;Lee, Byung-Moo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.113-113
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    • 2017
  • Downy mildew (DM), caused by several species in the Peronosclerospora and Scleropthora genera, is a major maize (Zea mays L.) disease in tropical or subtropical regions. DM is an obligate parasite species in the higher plants and spreads by oospores, wind, and mycelium in seed surface, soil, and living hosts. Owing to its geographical distribution and destructive yield reduction, DM is one of the most severe maize diseases among the maize pathogens. Positional cloning in combination with phenotyping is a general approach to identify disease resistant gene candidates in plants; however, it requires several time-consuming steps including population or fine mapping. Therefore, in the present study, we suggest a new combination strategy to improve the identification of disease resistant gene candidates. Downy mildew (DM) resistant maize was selected from five cultivars using the spreader row technique. Positional cloning and bioinformatics tools identified the DM resistant QTL marker (bnlg1702) and 47 protein coding genes annotations. Eventually, 5 DM resistant gene candidates, including bZIP34, Bak1, and Ppr, were identified by quantitative RT-PCR without fine mapping of the bnlg1702 locus. Specifically, we provided DM resistant gene candidates with our new strategy, including field selection by the spreader row technique without population preparation, the DM resistance region identification by positional cloning using bioinformatics tools, and expression level profiling by quantitative RT-PCR without fine mapping. As whole genome information is available for other crops, we propose applying our novel protocol to other crops or for other diseases with suitable adjustment.

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Bayesian Model Selection for Linkage Analyses: Considering Collinear Predictors (연관분석을 위한 베이지안 모형 선택: 상호상관성 변수를 중심으로)

  • Suh, Young-Ju
    • The Korean Journal of Applied Statistics
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    • v.18 no.3
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    • pp.533-541
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    • 2005
  • We identify the correct chromosome and locate the corresponding markers close to the QTL in the linkage analysis of a quantitative trait by using the SSVS method. We consider several markers linked to the QTL, as well as to each oyher and thus the i.b.d. values at these loci generate collinear predictors to be evaluated when using the SSVS approach. The results on considering only closely linked markers to two QTL simultaneously showed clear evidence in favor of the closest marker to the QTL considered over other markers. The results of the analysis of collinear markers with SSVS showeed high concordance to those obtained using traditional multiple regression. We conclude based on this simulation study that the SSVS is quite useful to identify linkage with multiple linked markers simultaneously for a complex quantitative trait.

Association between the Polymorphism in FUT1 Gene and the Resistance to PWD and ED in Three Pig Breeds

  • Luo, Yanru;Qiu, Xiaotian;Li, Hejun;Zhang, Qin
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.10
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    • pp.1268-1275
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    • 2010
  • Post-weaning diarrhoea (PWD) and oedema disease (ED) caused by E. coli F18 always result in economic losses to pig producers, and no effective methods of controlling PWD and ED are presently available. FUT1 has been identified as a candidate gene controlling the expression of E. coli F18 receptor. This study examined the correlation between F18ab and F18ac adhesion phenotypes and the polymorphism at position M307 of the FUT1 gene in three pig breeds (231 Large White, 107 Landrace and 109 Songliao Black). The results showed: i) Both the susceptible genotypes (GG and GA) and the adhesion phenotypes (adhesive or weekly adhesive) were dominant in all three breeds with frequencies over 95%. ii) Three adhesion patterns of the two F18 variants F18ab and F18ac, i.e., ($ab^+$, $ac^+$), ($ab^+$, $ac^-$) and ($ab^-$, $ac^-$), were found in all three breeds, and there was no significant difference in the distribution of adhesion phenotypes of the two variants (separately or jointly) among the three breeds (p>0.05). iii) The FUT1 M307 genotypes were completely associated with the F18ab adhesion phenotypes and very strongly associated with the F18ac adhesion phenotypes. All individuals of genotype AA were non-adhesive to both F18ab and F18ac. All individuals of genotype GG or GA were adhesive to F18ab, whereas 11% of them were non-adhesive to F18ac. These results suggest that the polymorphism at FUT1 M307 can be used for marker-assisted selection of PWD and ED resistant pigs.

Evaluation of BTA1 and BTA5 QTL Regions for Growth and Carcass Traits in American and Korean Cattle

  • Kim, K.S.;Kim, S.W.;Raney, N.E.;Ernst, C.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.11
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    • pp.1521-1528
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    • 2012
  • Previously identified QTL regions on BTA1 and BTA5 were investigated to validate the QTL regions and to identify candidate genes for growth and carcass traits in commercial cattle populations from the USA and Korea. Initially, a total of 8 polymorphic microsatellite (MS) markers in the BTA1 and 5 QTL regions were used for Chi-square tests to compare the frequencies of individual alleles between high and low phenotypic groups for the US (Michigan Cattleman's Association/Michigan State University; MCA/MSU) cattle. For a subsequent study, 24 candidate genes containing missense mutations and located within the QTL regions based on bovine genome sequence data were analyzed for genotyping in the two commercial cattle populations. Re-sequencing analyses confirmed 18 public missense SNPs and identified 9 new SNPs. Seventeen of these SNPs were used for genotyping of the MCA/MSU cattle (n = 98) and Korean native cattle (n = 323). On BTA1, UPK1B, HRG, and MAGEF1 polymorphisms residing between BM1312 and BMS4048 were significantly associated with growth and carcass traits in one or both of the MCA/MSU and Korean populations. On BTA5, ABCD2, IL22 and SNRPF polymorphisms residing between BL4 and BR2936 were associated with marbling and backfat traits in one or both of the MCA/MSU and Korean cattle populations. These results suggested that BTA 1 and 5 QTL regions may be segregating in both Korean Hanwoo and USA commercial cattle populations and DNA markers tested in this study may contribute to the identification of positional candidate genes for marker-assisted selection programs.

A SNP Harvester Analysis to Better Detect SNPs of CCDC158 Gene That Are Associated with Carcass Quality Traits in Hanwoo

  • Lee, Jea-Young;Lee, Jong-Hyeong;Yeo, Jung-Sou;Kim, Jong-Joo
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.6
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    • pp.766-771
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    • 2013
  • The purpose of this study was to investigate interaction effects of genes using a Harvester method. A sample of Korean cattle, Hanwoo (n = 476) was chosen from the National Livestock Research Institute of Korea that were sired by 50 Korean proven bulls. The steers were born between the spring of 1998 and the autumn of 2002 and reared under a progeny-testing program at the Daekwanryeong and Namwon branches of NLRI. The steers were slaughtered at approximately 24 months of age and carcass quality traits were measured. A SNP Harvester method was applied with a support vector machine (SVM) to detect significant SNPs in the CCDC158 gene and interaction effects between the SNPs that were associated with average daily gains, cold carcass weight, longissimus dorsi muscle area, and marbling scores. The statistical significance of the major SNP combinations was evaluated with $x^2$-statistics. The genotype combinations of three SNPs, g.34425+102 A>T(AA), g.4102636T>G(GT), and g.11614-19G>T(GG) had a greater effect than the rest of SNP combinations, e.g. 0.82 vs. 0.75 kg, 343 vs. 314 kg, 80.4 vs $74.7cm^2$, and 7.35 vs. 5.01, for the four respective traits (p<0.001). Also, the estimates were greater compared with single SNPs analyzed (the greatest estimates were 0.76 kg, 320 kg, $75.5cm^2$, and 5.31, respectively). This result suggests that the SNP Harvester method is a good option when multiple SNPs and interaction effects are tested. The significant SNPs could be applied to improve meat quality of Hanwoo via marker-assisted selection.

Validation of Single Nucleotide Polymorphisms Associated with Carcass Traits in a Commercial Hanwoo Population

  • Sudrajad, Pita;Sharma, Aditi;Dang, Chang Gwon;Kim, Jong Joo;Kim, Kwan Suk;Lee, Jun Heon;Kim, Sidong;Lee, Seung Hwan
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.11
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    • pp.1541-1546
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    • 2016
  • Four carcass traits, namely carcass weight (CW), eye muscle area (EMA), back fat thickness (BF), and marbling score (MS), are the main price decision parameters used for purchasing Hanwoo beef. The development of DNA markers for these carcass traits for use in a beef management system could result in substantial profit for beef producers in Korea. The objective of this study was to validate the association of highly significant single nucleotide polymorphisms (SNPs) identified in a previous genome-wide association study (GWAS) with the four carcass traits in a commercial Hanwoo population. We genotyped 83 SNPs distributed across all 29 autosomes in 867 steers from a Korean Hanwoo feedlot. Six SNPs, namely ARS-BFGL-NGS-22774 (Chr4, Pos:4889229), ARS-BFGL-NGS-100046 (Chr6, Pos:61917424), ARS-BFGL-NGS-39006 (Chr27, Pos:38059196), ARS-BFGL-NGS-18790 (Chr10, Pos:26489109), ARS-BFGL-NGS-43879 (Chr9, Pos:39964297), and BTB-00775794 (Chr20, Pos:20476265), were found to be associated with CW, EMA, BF, and MS. The ARS-BFGL-NGS-22774, BTB-00775794, and ARS-BFGL-NGS-39006 markers accounted for 1.80%, 1.72%, and 1.35% (p<0.01), respectively, of the phenotypic variance in the commercial Hanwoo population. Many genes located in close proximity to the significant SNPs identified in this study were previously reported to have roles in carcass traits. The results of this study could be useful for marker-assisted selection programs.

Genetic Variation of Growth Hormone Gene and Its Relationship with Milk Production Traits in China Holstein Cows

  • Zhou, Guo-li;Zhu, Qi;Jin, Hai-guo;Guo, Shan-li
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.315-318
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    • 2006
  • Associations were analysed between polymorphisms localized in intron 3 of the growth hormone gene (GH-MspI) and milk production traits of 543 China Holstein cows. A PCR-RFLP method was used for identification of genotypes. The following frequencies of genotypes and alleles were found: 0.77, 0.21 and 0.02 for +/+, +/- and -/-, respectively, and 0.87 and 0.13 for $GH^+$ and $GH^-$, respectively. Significant differences between herds were observed in the frequencies of both genotypes and alleles. The results of least squares analysis showed that in all three lactation phases the GH +/+ cows yielded most milk (p<0.01 for lactation I and p<0.05 for lactations II and III), whereas +/- cows showed higher milk fat content than +/+ individuals (p<0.05 for lactation I and II, and p<0.01 for lactation III). The +/+ cows yielded more fat than +/- individuals (p<0.01 only in lactation I). The +/+ cows yielded more milk protein than +/- individuals (p<0.01 for lactation I, II, and III). The +/+ cows produced milk of higher protein content than that of +/- individuals (p<0.05 only in lactation II). Based on these results, we conclude that the +/+ of GH locus should be the favored genotype in China Holstein cow breeds for use in marker-assisted selection programmes.

Identification of Single Nucleotide Polymorphisms (SNPs) of the Bovine Growth Hormone (bGH) Gene Associated with Growth and Carcass Traits in Hanwoo

  • Lee, Ji-Hong;Lee, Yun-Mi;Lee, Jea-Young;Oh, Dong-Yep;Jeong, Dae-Jin;Kim, Jong-Joo
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.10
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    • pp.1359-1364
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    • 2013
  • The purpose of this study was to find any association of the bovine growth hormone (bGH) gene with growth and carcass quality traits in Korean native cattle, Hanwoo. Genomic DNA was extracted from 21 Hanwoo individuals, and the 47 to 2,528 bp region of the bGH 2,856 bp (GenBank accession number M57764) including the promoter and the five exons was sequenced. A total of ten bGH SNPs were confirmed, including four (253 C>T, 303 C>T, 502 C>T, and 559 G>A) in the promoter, one (679 C>T) in exon 1, one (1,692 T>C) in intron 3, and four (2141 C>G, 2258 C>T, 2277 C>T, and 2291 A>C) in exon 5. The ten bGH SNPs were genotyped for a sample of 242 Hanwoo steers and association tests were performed to find any significant SNP that was correlated with growth and carcass quality. Of the SNPs, the 303 C>T SNP in the promoter region was significantly associated with 6-month-old weight, the 559 G>A SNP with longissimus dorsi muscle area, the 2141 C>G SNP in exon 5 with daily weight gain, and the 2258 C>T SNP with daily weight gain and carcass weight (p<0.05). The significant SNPs need to be verified in other Hanwoo populations before considering implementation of marker-assisted selection for genetic improvement of growth and carcass quality in Hanwoo.