• Title/Summary/Keyword: marker-assisted breeding

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A New Variety "Singang" with resistance to Soybean Mosaic Virus by Molecular Marker Assisted Selection (분자표지선발(MAS)을 이용한 콩모자이크병 저항성 신품종 "신강")

  • Lee, Seuk-ki;Moon, Jung-Kyung;Jeong, Kwang-Ho;Seo, Min-Jung;Kim, Yul-Ho;Yun, Hong-Tae;Kim, Jeong-Kon
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.568-573
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    • 2009
  • A new soybean variety, "Singang", was released by NICS, RDA in 2008. It is resistant to soybean mosic virus (SMV), which is one of the most serious diseases in soybean production. "Singang" had been developed for seven years by molecular marker-assisted selection (MAS) breeding method using recurrent parent, "Sowon", and donor parent, "L29" which has the SMV-resistance gene, Rsv3. Selected promising breeding line, Suwon 242, was evaluated in yield trials for 2 years from 2005 to 2006 at NICS. Regional yield trials were performed for 3 years from 2006 to 2008 in five locations including Jeju. Agronomic characteristics of "Singang" is a determinate growth habit, purple flower color, grey pubescence, yellow testa, light brown hilum and lanceolate leaflet shape. "Singang" showed the similar performance to "Sowon" cultivar. This variety showed 3 days shorter in days to maturity than the check variety (Pungsannamul). The average yield under regional yield trials in five location was 2.78t/ha. It is adaptable to the whole country except for Gangwon province.

Mapping of grain alkali digestion trait using a Cheongcheong/Nagdong doubled haploid population in rice

  • Kim, Hak Yoon;Kim, Kyung-Min
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.76-81
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    • 2016
  • We performed a molecular marker-based analysis of quantitative trait loci for traits that determine the quality of appearance of grains using 120 doubled haploid lines developed by anther culture from the F1 cross between 'Cheongcheong' (Oryza sativa L. ssp. Indica) and 'Nagdong' (Oryza sativa L. ssp. Japonica). We therefore calculated the alkali digestion value (ADV), used to indirectly measure gelatinization temperature, to evaluate the quality of cooked rice in 2013 and 2014. The ADV score of frequency distribution was higher milled rice than brown rice. In total, nine different quantitative trait loci (QTLs) were found on 5 chromosomes in 2013 and 2014. Also, chromosome 5, 8 were detected over two years. We conclude that selected molecular markers from this QTL analysis could be exploited in future rice quality. In conclusion, we investigated ADV of brown and milled rice in CNDH population. This study found nine QTLs related to the ADV of brown and milled rice. The detected one marker can be used to select lines with desirable eating-quality traits because ADV is closely associated with the eating quality of cooked rice. Therefore, it will be useful to collect resources and distinguishable in many varieties for rice breeding program.

Molecular discrimination of Panax ginseng cultivar K-1 using pathogenesis-related protein 5 gene

  • Wang, Hongtao;Xu, Fengjiao;Wang, Xinqi;Kwon, Woo-Saeng;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.43 no.3
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    • pp.482-487
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    • 2019
  • Background: The mixed-cultivation of different Panax ginseng cultivars can cause adverse effects on stability of yield and quality. K-1 is a superior cultivar with good root shape and stronger disease resistance. DNA markers mined from functional genes are clearly desirable for K-1, as they may associate with major traits and can be used for marker-assisted selection to maintain the high quality of Korean ginseng. Methods: Five genes encoding pathogenesis-related (PR) proteins of P. ginseng were amplified and compared for polymorphism mining. Primary, secondary, and tertiary structures of PR5 protein were analyzed by ExPASy-ProtParam, PSSpred, and I-TASSER methods, respectively. A coding single nucleotide polymorphism (SNP)-based specific primer was designed for K-1 by introducing a destabilizing mismatch within the 3' end. Allele-specific polymerase chain reaction (PCR) and real-time allele-specific PCR assays were conducted for molecular discrimination of K-1 from other cultivars and landraces. Results: A coding SNP leading to the modification of amino acid residue from aspartic acid to asparagine was exploited in PR5 gene of K-1 cultivar. Bioinformatics analysis showed that the modification of amino acid residue changed the secondary and tertiary structures of the PR5 protein. Primer KSR was designed for specific discrimination of K-1 from other ginseng cultivars and landraces. The developed real-time allele-specific PCR assay enabled easier automation and accurate genotyping of K-1 from a large number of ginseng samples. Conclusion: The SNP marker and the developed real-time allele-specific PCR assay will be useful not only for marker-assisted selection of K-1 cultivar but also for quality control in breeding and seed programs of P. ginseng.

Molecular Screening of Blast Resistance Genes in Rice using SSR Markers

  • Singh, A.K.;Singh, P.K.;Arya, Madhuri;Singh, N.K.;Singh, U.S.
    • The Plant Pathology Journal
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    • v.31 no.1
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    • pp.12-24
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    • 2015
  • Rice Blast is the most devastating disease causing major yield losses in every year worldwide. It had been proved that using resistant rice varieties would be the most effective way to control this disease. Molecular screening and genetic diversities of major rice blast resistance genes were determined in 192 rice germplasm accessions using simple sequence repeat (SSR) markers. The genetic frequencies of the 10 major rice blast resistance genes varied from 19.79% to 54.69%. Seven accessions IC337593, IC346002, IC346004, IC346813, IC356117, IC356422 and IC383441 had maximum eight blast resistance gene, while FR13B, Hourakani, Kala Rata 1-24, Lemont, Brown Gora, IR87756-20-2-2-3, IC282418, IC356419, PKSLGR-1 and PKSLGR-39 had seven blast resistance genes. Twenty accessions possessed six genes, 36 accessions had five genes, 41 accessions had four genes, 38 accessions had three genes, 26 accessions had two genes, 13 accessions had single R gene and only one accession IC438644 does not possess any one blast resistant gene. Out of 192 accessions only 17 accessions harboured 7 to 8 blast resistance genes.

Development of Fluidigm SNP Type Genotyping Assays for Marker-assisted Breeding of Chili Pepper (Capsicum annuum L.)

  • Kim, Haein;Yoon, Jae Bok;Lee, Jundae
    • Horticultural Science & Technology
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    • v.35 no.4
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    • pp.465-479
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    • 2017
  • Chili pepper (Capsicum annuum L.) is an economically important horticultural crop in Korea; however, various diseases, including Phytophthora root rot, anthracnose, powdery mildew, Cucumber mosaic virus (CMV), Pepper mild mottle virus (PMMoV), and Pepper mottle virus (PepMoV), severely affect their productivity and quality. Therefore, pepper varieties with resistance to multiple diseases are highly desired. In this study, we developed 20 SNP type assays for three pepper populations using Fluidigm nanofluidic dynamic arrays. A total of 4,608 data points can be produced with a 192.24 dynamic array consisting of 192 samples and 24 SNP markers. The assays were converted from previously developed sequence-tagged-site (STS) markers and included markers for resistance to Phytophthora root rot (M3-2 and M3-3), anthracnose (CcR9, CA09g12180, CA09g19170, CA12g17210, and CA12g19240), powdery mildew (Ltr4.1-40344, Ltr4.2-56301, and Ltr4.2-585119), bacterial spot (Bs2), CMV (Cmr1-2), PMMoV (L4), and PepMoV (pvr1 and pvr2-123457), as well as for capsaicinoids content (qcap3.1-40134, qcap6.1-299931, qcap6.1-589160, qdhc2.1-1335057, and qdhc2.2-43829). In addition, 11 assays were validated through a comparison with the corresponding data of the STS markers. Furthermore, we successfully applied the assays to commercial $F_1$ cultivars and to our breeding lines. These 20 SNP type assays will be very useful for developing new superior pepper varieties with resistance to multiple diseases and a higher content of capsaicinoids for increased pungency.

The Prediction Ability of Genomic Selection in the Wheat Core Collection

  • Yuna Kang;Changsoo Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.235-235
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    • 2022
  • Genome selection is a promising tool for plant and animal breeding, which uses genome-wide molecular marker data to capture large and small effect quantitative trait loci and predict the genetic value of selection candidates. Genomic selection has been shown previously to have higher prediction accuracies than conventional marker-assisted selection (MAS) for quantitative traits. In this study, the prediction accuracy of 10 agricultural traits in the wheat core group with 567 points was compared. We used a cross-validation approach to train and validate prediction accuracy to evaluate the effects of training population size and training model.As for the prediction accuracy according to the model, the prediction accuracy of 0.4 or more was evaluated except for the SVN model among the 6 models (GBLUP, LASSO, BayseA, RKHS, SVN, RF) used in most all traits. For traits such as days to heading and days to maturity, the prediction accuracy was very high, over 0.8. As for the prediction accuracy according to the training group, the prediction accuracy increased as the number of training groups increased in all traits. It was confirmed that the prediction accuracy was different in the training population according to the genetic composition regardless of the number. All training models were verified through 5-fold cross-validation. To verify the prediction ability of the training population of the wheat core collection, we compared the actual phenotype and genomic estimated breeding value using 35 breeding population. In fact, out of 10 individuals with the fastest days to heading, 5 individuals were selected through genomic selection, and 6 individuals were selected through genomic selection out of the 10 individuals with the slowest days to heading. Therefore, we confirmed the possibility of selecting individuals according to traits with only the genotype for a shorter period of time through genomic selection.

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Developmental Changes of Recessive Genes-mediated Cucumber mosaic virus (CMV) Resistance in Peppers (Capsicum annuum L.)

  • Min, Woong-Ki;Ryu, Jae-Hwang;Ahn, Su-Hyeon
    • Horticultural Science & Technology
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    • v.32 no.2
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    • pp.235-240
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    • 2014
  • Cucumber mosaic virus (CMV) is one of the most important viral diseases in pepper (Capsicum annuum L.), and several genes for resistance were reported in Capsicum spp. In Korea, a single dominant gene that is resistant to $CMV_{Fny}$ and $CMV_{P0}$ has been used for breeding. Recently, a new strain ($CMV_{P1}$) was reported that could infect cultivars resistant to both $CMV_{Fny}$ and $CMV_{P0}$. Therefore, breeding of more robust CMV-resistant cultivars is required. In this study, we surveyed the inheritance of $CMV_{P1}$ resistance and analyzed the location of the resistance loci. After $CMV_{P1}$ inoculation of various germplasms and breeding lines, one accession (ICPN18-8) showed no visual symptoms at 15 dpi (days post inoculation) but was susceptible after 45 dpi, and one resistant line (I7339) showed resistance until at 45 dpi. The latter line was used for tests of resistance inheritance. A total of 189 $F_2$ plants were examined, with 42 individuals showing resistance at 15 dpi and a phenotype segregation ratio close to 1:3 (resistant:susceptible plants). In a lateral ELISA test at 45 dpi, 11 plants showed resistance, and the segregation ratio was changed to 1:15. These results indicate that resistance in C. annuum 'I7339' is controlled by two different recessive genes; we named these resistance genes 'cmr3E' and 'cmr3L,' respectively. To locate these two resistant loci in the pepper linkage map, various RAPD, SSR, and STS markers were screened; only nine markers were grouped into one linkage group (LG). Only one RAPD primer (OPAT16) was distantly linked with cmr3E (22.3 cM) and cmr3L (20.7 cM). To develop more accurate markers for marker-assisted breeding, enriching for molecular markers spanning two loci will be required.

Effect of RFLP Marker of the Mitochondrial DNA D-Loop Region on Milk Production in Korean Cattle (한우 Mitochondrial DNA D-Loop 영역의 RFLP Marker가 산유량에 미치는 영향)

  • Chung Eui-Ryong;Chung Ku-Young
    • Food Science of Animal Resources
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    • v.25 no.2
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    • pp.218-225
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    • 2005
  • This study was performed to determine sequence variation and RFLP of the mt DNA D-loop region using Southern blot hybridization analysis and to develop mt DNA marker affecting milk production traits in Hanwoo cows. The PCR was used to amplify an 1142 bp fragment within the D-loop region of mt DNA using specific primers. Mt DNA were digested with seven restriction enzymes and hybridized using DIG-labeled D-loop probe. The mt DNA RFLP polymorphisms were observed in the four enzymes, BamHI, RsaI, XbaI and HpaII. Nucleotide substitutions were detected at positions 441 (G/C), 469 (T/C), 503 (C/T), 569 (G/A), 614 (C/A) and 644 (C/T) of the mt DNA D-loop region between two selected lines. Significant relationship between the XbaI RFLP type and breeding value was found(p<0.05). Cows with A type had higher estimated breeding values than those with B type (P<0.05) between high and low milk production lines. Therefore, the RFLP marker of mt DNA could be used as a selection assisted tool for individuals with high milk producing ability in Hanwoo.

Development of Selectable Marker of High Oleate Trait in Peanut (Arachis hypogaea L.) (땅콩에서 고 올레인산 형질관련 분자마커의 선발)

  • Yang, Kiwoung;Pae, Suk-Bok;Park, Chang-Hwan;Lee, Myoung Hee;Jung, Chan-Sik;Son, Jeong-Hee;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.507-514
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    • 2010
  • Peanut(Arachis hypogaea L.) is one of the major oilseed crops. The peanut oil consists of palmitic, oleic and linoleic acids, which are present at levels of 10%, 36-67% and 15-43%, respectively. High oleate mutant of peanut F435 contains 80% oleate and as little as 2% linoleate in seed oil. Previous study indicated that delta 12 fatty acid desaturase is a major enzyme controlling the oleate content in seeds of oilseed crops. F435 sequence alignment of their coding regions disclosed that an extra A(adenine) was inserted at the position +2,823 bp of delta 12 fatty acid desaturase gene. This study was to develop molecular marker (SNP marker) co-segregating with the high oleate trait. Chopyeong ${\times}$ F435 $F_2$ 41 population were investigated using molecular marker and fatty acid assay (NIR and gas chromatography). Finally, this marker segregates Chopyeong type 26 lines, heterotype 9 lines and F435 type 6 lines. These results in our study suggested that SNP marker conform fatty acid assay.

Marker Assisted Selection of Brown Planthopper Resistance and Development of Multi-Resistance to Insect and Diseases in Rice (Oryza sativa L.) (DNA 마커를 이용한 벼멸구 저항성 선발 및 복합내병충성 벼 계통 육성)

  • Lee, Jong-Hee;Yeo, Un-Sang;Cho, Jun-Hyun;Lee, Ji-Yoon;Song, You-Chun;Shin, Mun-Sik;Kang, Hang-Won;Sohn, Jae-Keun
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.413-421
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    • 2011
  • The main objective of this study was to develop the multi-resistance lines to insects(brown planthopper; BPH, rice green leafhopper; GRH) and disease(blast; BL, bacterial blight; BB and rice stripe virus disease;RSV) with good grain quality and plant type by combining conventional breeding and marker assisted selection(MAS) and to eliminate the linkage drag effects between Bph1 gene and culm length, we conducted MAS of Bph1 gene in advanced backcross and double cross progenies. 'Nampyeong', 'Junam' and 'Milyang220' were used as the parent in this study. 'Milyang220' was used as the donor of brown planthopper resistance gene Bph1 with tall culm length. Two backcross progenies were developed using two recipients 'Nampyeong' carrying GRH resistance gene Grh3(t) with good grain appearance and 'Junam' harboring bacterial blight resistance gene Xa3 with short culm length. Two $BC_1$ generations were resulted from the backcrossing of the $F_1$ plants with recurrent parents 'Nampyeong' and 'Junam'. The second rounds of backcrossing($BC_2$) were derived from the cross of selected resistant $BC_1F_1$ plants based on heterozygous genotype of RM28493 linked to Bph1 gene. The double crossed population was constructed from the cross of between each heterozygous $BC_2F_1$ plants at RM28493 locus of '$Nampyeong^*3$ / Milyang220' and '$Junam^*3$ / Milyang220'., The homozygous alleles in Bph1 gene were selected using co-dominant DNA marker RM28493 in double crossed population. Eighty-five lines with multi-resistance to BL, BB, RSV, GRH and BPH were selected by bio-assay and MAS in generation of double crossing. The culm length, head rice ratio and yield of the selected multi resistance lines was ranged from 71 to 88 cm, from 51 to 93%, from 449 to 629 kg/10a. respectively. We can select a promising multi resistance line similar with 'Nampyeong' of major agronomic traits such as culm legnth, head rice ratio and yield. It was designated as Milyang265. Finally this study was developed the multi resistant varieties against to insects and diseases with the good grain quality 'Milyang265' by the advanced backcross and double cross combining MAS and it can be used as genetic resources of multi-resistance to insect and diseases in rice breeding programs.