• Title/Summary/Keyword: Marker selection

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Development of a Sequence Characteristic Amplified Region Marker linked to the L4 Locus Conferring Broad Spectrum Resistance to Tobamoviruses in Pepper Plants

  • Kim, Hyun Jung;Han, Jung-Heon;Yoo, Jae Hyoung;Cho, Hwa Jin;Kim, Byung-Dong
    • Molecules and Cells
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    • v.25 no.2
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    • pp.205-210
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    • 2008
  • To develop molecular markers linked to the $L^4$ locus conferring resistance to tobamovirus pathotypes in pepper plants, we performed AFLP with 512 primer combinations for susceptible (S pool) and resistant (R pool) DNA bulks against pathotype 1.2 of pepper mild mottle virus. Each bulk was made by pooling the DNA of five homozygous individuals from a T10 population, which was a near-isogenic $BC_4F_2$ generation for the $L^4$ locus. A total of 19 primer pairs produced scorable bands in the R pool. Further screening with these primer pairs was done on DNA bulks from T102, a $BC_{10}F_2$ derived from T10 by back crossing. Three AFLP markers were finally selected and designated L4-a, L4-b and L4-c. L4-a and L4-c each underwent one recombination event, whereas no recombination for L4-b was seen in 20 individuals of each DNA bulk. Linkage analysis of these markers in 112 $F_2$ T102 individuals showed that they were each within 2.5 cM of the $L^4$ locus. L4-b was successfully converted into a simple 340-bp SCAR marker, designated L4SC340, which mapped 1.8 cM from the $L^4$ locus in T102 and 0.9 cM in another $BC_{10}F_2$ population, T101. We believe that this newly characterized marker will improve selection of tobamovirus resistance in pepper plants by reducing breeding cost and time.

Predicting the Accuracy of Breeding Values Using High Density Genome Scans

  • Lee, Deuk-Hwan;Vasco, Daniel A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.2
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    • pp.162-172
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    • 2011
  • In this paper, simulation was used to determine accuracies of genomic breeding values for polygenic traits associated with many thousands of markers obtained from high density genome scans. The statistical approach was based upon stochastically simulating a pedigree with a specified base population and a specified set of population parameters including the effective and noneffective marker distances and generation time. For this population, marker and quantitative trait locus (QTL) genotypes were generated using either a single linkage group or multiple linkage group model. Single nucleotide polymorphism (SNP) was simulated for an entire bovine genome (except for the sex chromosome, n = 29) including linkage and recombination. Individuals drawn from the simulated population with specified marker and QTL genotypes were randomly mated to establish appropriate levels of linkage disequilibrium for ten generations. Phenotype and genomic SNP data sets were obtained from individuals starting after two generations. Genetic prediction was accomplished by statistically modeling the genomic relationship matrix and standard BLUP methods. The effect of the number of linkage groups was also investigated to determine its influence on the accuracy of breeding values for genomic selection. When using high density scan data (0.08 cM marker distance), accuracies of breeding values on juveniles were obtained of 0.60 and 0.82, for a low heritable trait (0.10) and high heritable trait (0.50), respectively, in the single linkage group model. Estimates of 0.38 and 0.60 were obtained for the same cases in the multiple linkage group models. Unexpectedly, use of BLUP regression methods across many chromosomes was found to give rise to reduced accuracy in breeding value determination. The reasons for this remain a target for further research, but the role of Mendelian sampling may play a fundamental role in producing this effect.

Prediction of genomic breeding values of carcass traits using whole genome SNP data in Hanwoo (Korean cattle) (한우에 있어서 유전체 육종가 추정)

  • Lee, Seung Hwan;Kim, Heong Cheul;Lim, Dajeong;Dang, Chang Gwan;Cho, Yong Min;Kim, Si Dong;Lee, Hak Kyo;Lee, Jun Heon;Yang, Boh Suk;Oh, Sung Jong;Hong, Seong Koo;Chang, Won Kyung
    • Korean Journal of Agricultural Science
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    • v.39 no.3
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    • pp.357-364
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    • 2012
  • Genomic breeding value (GEBV) has recently become available in the beef cattle industry. Genomic selection methods are exceptionally valuable for selecting traits, such as marbling, that are difficult to measure until later in life. One method to utilize information from sparse marker panels is the Bayesian model selection method with RJMCMC. The accuracy of prediction varies between a multiple SNP model with RJMCMC (0.47 to 0.73) and a least squares method (0.11 to 0.41) when using SNP information, while the accuracy of prediction increases in the multiple SNP (0.56 to 0.90) and least square methods (0.21 to 0.63) when including a polygenic effect. In the multiple SNP model with RJMCMC model selection method, the accuracy ($r^2$) of GEBV for marbling predicted based only on SNP effects was 0.47, while the $r^2$ of GEBV predicted by SNP plus polygenic effect was 0.56. The accuracies of GEBV predicted using only SNP information were 0.62, 0.68 and 0.73 for CWT, EMA and BF, respectively. However, when polygenic effects were included, the accuracies of GEBV were increased to 0.89, 0.90 and 0.89 for CWT, EMA and BF, respectively. Our data demonstrate that SNP information alone is missing genetic variation information that contributes to phenotypes for carcass traits, and that polygenic effects compensate genetic variation that whole genome SNP data do not explain. Overall, the multiple SNP model with the RJMCMC model selection method provides a better prediction of GEBV than does the least squares method (single marker regression).

Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains with higher β-glucan (베타글루칸 함량이 높은 큰느타리버섯 선발을 위한 SCAR marker 개발)

  • Kim, Su Cheol;Kim, Hye Soo;Cho, Yong Un;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.79-83
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    • 2015
  • In this study, SCAR marker that differentiates Pleurotus eryngii strains with higher ${\beta}$-glucan from control strain was developed. Genomic DNAs of 9 control strains of Pleurotus eryngii and 9 Pleurotus eryngii strains with higher ${\beta}$-glucan were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). One-hundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 91 bp was yielded by OP-R03 primer from the Pleurotus eryngii strains with higher ${\beta}$-glucan. A sequence characterized amplified region (SCAR) marker, designated as OP-R03-1-F and OP-R03-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-R03-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains with higher ${\beta}$-glucan from the control strains.

Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains adaptable to high-temperature (큰느타리버섯의 고온적응성 형질에 관련된 SCAR Marker 개발)

  • Kim, Su Cheol;Kim, Hye Soo;Park, So Yeon;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.226-231
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    • 2014
  • In this study, SCAR marker that differentiates Pleurotus eryngii strains adaptable to high-temperature from control strain was developed. Genomic DNAs of 7 control strains of Pleurotus eryngii and 7 Pleurotus eryngii strains adaptable to high-temperature were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). Onehundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 385 bp was yielded by OP-A06 primer from the Pleurotus eryngii strains adaptable to high-temperature. A sequence characterized amplified region (SCAR) marker, designated as OP-A06-1-F and OP-A06-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-A06-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains adaptable to high-temperature from the control strains.

System for Repeated Integration of Various Gene Expression Cassettes in the Yeast Chromosome (효모염색체내에 다양한 유전자발현 cassette의 반복적 integration을 위한 system 구축)

  • Kim, Yeon-Hee
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1277-1284
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    • 2018
  • In this study, a repeated yeast integrative plasmid (R-YIp) harboring Cre/loxP system was constructed to integrate various gene expression cassettes into the yeast chromosome. The R-YIp system contains a reusable selective marker (CgTRP1), loxP sequence, and target sequence for integration. Therefore, many gene expression cassettes can be integrated into the same position of the same yeast chromosome. In the present study, several model enzymes involving xylan/xylose metabolism were examined, including endoxylanase (XYLP), ${\beta}$-xylosidase (XYLB), xylose reductase (GRE3) and xylitol dehydrogenase (XYL2). Efficient expression of these genes was obtained using two promoters (GAL10p and ADH1p) and various plasmids (pGMF-GENE and pAMF-GENE plasmids) were constructed. The XYLP, XYLB, GRE3, and XYL2 genes were efficiently expressed under the control of the GAL10 promoter. Subsequently, R-YIps containing the GAL10p-GENE-GAL7t cassette were constructed, resulting in pRS-XylP, pRS-XylB, pRS-Gre3, and pRS-Xyl2 plasmids. These plasmids were sequentially integrated into chromosome VII of a Saccharomyces cerevisiae strain by repeated gene integration and selective marker rescue. These genes were integrated by the R-YIp system and were stably expressed in the yeast transformants to produce active recombinant enzymes. Therefore, we expect that the R-YIp system will be able to overcome current limitations of the host cells and allow selective marker selection for the integration of various genes into the yeast chromosome.