• Title/Summary/Keyword: STS marker

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Analysis of Korean japonica rice cultivars using molecular markers associated with blast resistance genes

  • Suh, Jung-Pil;Roh, Jae-Hwan;Cho, Young-Chan;Han, Seong-Sook;Jeon, Yong-Hee;Kang, Kyung-Ho;Kim, Yeon-Gyu
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.215-222
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    • 2008
  • Fifty-two Korean japonica rice cultivars were analyzed for leaf blast resistance and genotyped with 4 STS and 26 SSR markers flanking the specific chromosome sites linked with blast resistance genes. In our analysis of resistance genes in 52 japonica cultivars using STS markers tightly linked to Pib, Pita, Pi5(t) and Pi9(t), the blast nursery reaction of the cultivars possessing the each four major genes were not identical to that of the differential lines. Eight of the 26 SSR markers were associated with resistant phenotypes against the isolates of blast nursery as well as the specific Korean blast isolates, 90-008 (KI-1113), 03-177 (KJ-105). These markers were linked to Pit, Pish, Pib, Pi5(t), Piz, Pia, Pik, Pi18, Pita and Pi25(t) resistance gene loci. Three of the eight SSR markers, MRG5836, RM224 and RM7102 only showed significantly associated with the phenotypes of blast nursery test for two consecutive years. These three SSR markers also could distinguish between resistant and susceptible japonica cultivars. These results demonstrate the usefulness of marker-assisted selection and genotypic monitoring for blast resistance of rice in blast breeding programs.

High-Throughput DNA Extraction Method for Marker Analysis in Rice Grain (대량의 쌀 시료 분석을 위한 DNA 추출법)

  • Choi, Young-Deok;Lee, Hae-Kwang;Lee, Yun-Suk;Yun, Jeong-Hee;Kim, Su-Jeong;Park, Seong-Whan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.269-273
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    • 2006
  • The study of molecular markers to improve crops largely depends on the availability of rapid and of efficient DNA extraction methods. Here we developed a cheap and convenient method to isolate genomic DNA from rice grains suitable for large-scale microsatellite analysis. We confirmed that the isolated rice DNA is suitable for PCR analysis with STS marker and SNP marker, as well as microsatellite marker. Further, we established high-throughput DNA extraction system in a 96-well plate format which make it possible high-throughput analysis of microsatellite markers with rice grains. This implies that the new method could be a useful tool for other types of marker analysis in large scale.

Development and Validation of a Perfect KASP Marker for Fusarium Head Blight Resistance Gene Fhb1 in Wheat

  • Singh, Lovepreet;Anderson, James A;Chen, Jianli;Gill, Bikram S;Tiwari, Vijay K;Rawat, Nidhi
    • The Plant Pathology Journal
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    • v.35 no.3
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    • pp.200-207
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    • 2019
  • Fusarium head blight (FHB) is a devastating wheat disease with a significant economic impact. Fhb1 is the most important large effect and stable QTL for FHB resistance. A pore-forming toxin-like (PFT) gene was recently identified as an underlying gene for Fhb1 resistance. In this study, we developed and validated a PFT-based Kompetitive allele specific PCR (KASP) marker for Fhb1. The KASP marker, PFT_KASP, was used to screen 298 diverse wheat breeding lines and cultivars. The KASP clustering results were compared with gelbased gene specific markers and the widely used linked STS marker, UMN10. Eight disagreements were found between PFT_KASP and UMN10 assays among the tested lines. Based on the genotyping and sequencing of genes in the Fhb1 region, these genotypes were found to be common with a previously characterized susceptible haplotype. Therefore, our results indicate that PFT_KASP is a perfect diagnostic marker for Fhb1 and would be a valuable tool for introgression and pyramiding of FHB resistance in wheat cultivars.

Construction of a Genetic Linkage Map in Radish(Raphanus sativus L.) Using RAPD Markers (RAPD 마커를 이용한 무의 유전자지도 작성)

  • Ahn, Choon-Hee;Choi, Su-Ryun;Lim, Yong-Pyo;Chung, Hae-Joon;Yae, Byeong-Woo;Yoon, Wha-Mo
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.151-159
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    • 2002
  • Genetic map and molecular marker have a great importance in improving and facilitating crop breeding program as well as in genome analysis and map-based cloning of genes representing desirable characters. This study aimed at developing RAPD markers and constructing a genetic linkage map using 82 BC$_1$F$_1$individuals originated from the cross between '835' and B$_2$in radish (Raphanus sativus L.). One of the parents for genetic linkage map construction, '835'(P$_1$) of egg type is susceptible to Fusarium wilt and have medium resistance to virus infection and the other parent, B$_2$(P$_2$) of round type, is susceptible to Fusarium wilt and virus, Screening of 394 RAPD primers in BC$_1$F$_1$) population resulted in selecting 128 polymorphic markers which displayed 1:1 segregation pattern. Two markers failed to display 1:1 segregation and showed the segregation ratio skewed to maternal genotype. Selected markers were categorized into 14 linkage group based on LOD score represented by MAPMAKER/EXP program. Five groups composed of single marker among them were excluded from the linkage map, and consequently, the remaining groups are well matched with the number of radish chromosome (n=9). The linkage map constructed with 128 markers covers 1,688.3 cM and the average distance between markers was 13.8 cM. For developing STS marker, we determined the partial nucleotide sequence of OPE10 marker at both ends and designed a oligonucleotide primer pair based on this sequence. STS PCR using the primer pair displayed a single, clear band of which segregation is perfectly matched with that of OPE10 marker. This implies that RAPD markers could readily convert into clear and reliable STS markers.

Identification of Subspecies-specific STS Markers and Their Association with Segregation Distortion in Rice(Oryza sativa L.)

  • Chin, Joong-Hyoun;Kim, Jung-Hee;Jiang, Wenzhu;Chu, Sang-Ho;Woo, Mi-Ok;Han, Longzhi;Brar, Darshan;Koh, Hee-Jong
    • Journal of Crop Science and Biotechnology
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    • v.10 no.3
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    • pp.175-184
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    • 2007
  • Two subspecies, japonica and indica, have been reported in rice, which differ in several ecotypic traits. However, reproductive barriers in hybrid progenies between subspecies have been major obstacles in breeding programs using inter-subspecific hybridization. As the first step to elucidate the reproductive barriers, we developed subspecies-specific(SS) STS markers in this study. A total of 765 STS primers were designed through comparing DNA sequences at every $2{\sim}3$cM interval between japonica and indica rices, which are available at Web DBs such as IRGSP, NCBI, TIGR, and GRAMENE, and tested for subspecies-specificity using 15 indica and 15 japonica varieties of diverse origin. Of them, 67 STS markers were identified as SS STS markers and their subspecies-specificity scores were estimated. The SS markers were dispersed throughout the genome along chromosomes. Of them, 64 SS markers were mapped on an RIL population derived from a Dasanbyeo(indica)/TR22183(japonica) cross. Genomic inclination of RILs was evaluated based on the genotyping with different types of markers. Association test between markers and segregation distortion revealed that segregation distortion might not be the cause of generating SS markers. The SS markers will be applicable to estimate the genomic inclination of varieties or lines and to study the differentiation of indica and japonica, and ultimately to breed true hybrid rice varieties in which desirable characters from both subspecies are recombined.

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Screening of Resistance Genes Linked to Brown Planthopper Using STS Marker in Aromatic Rice Germplasm

  • Kim, Jeong-Soon;Ahn, Sang-Nag;Hong, Sung-Jun;Park, Jong-Ho;Lee, Min-Ho;Han, Eun-Jung;Damodaran, P.N.;Kim, Yong-Ki;Jee, Hyeong-Jin;Shim, Chang-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.2
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    • pp.167-176
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    • 2011
  • Brown planthopper (BPH) is a serious insect pest of rice crop throughout rice growing countries, and yield loss due to its infection can be up to 60%. This study aimed to evaluate efficiency of molecular markers for screening BPH resistance accessions among 86 aromatic rice germplasm Eighty-six accessions of aromatic rice germplasm included two accessions of Tongil type (bred in Korea), 28 accessions of japonica type and 56 accessions of indica type. We applied eight STS markers (pBPH9, pBPH19, pBPH20, pBPH21, AJ09-b, RG457L, RG457B, and 7312.T4A) which were linked to four of BPH resistance genes, Bph1, Bph13(t), Bph10, and Bph18(t) respectively. One japonica type accession, 415XIr352, and six indica type accessions possessed one or four positive bands when tested with four STS markers linked to Bph1 gene. One indica type aromatic rice, Basmati9-93, showed the target bands linked to the Bph10 gene. The other accessions did not show same fragments as the respective resistant lines. Bph13(t) is the most widely introduced resistance gene and only one accession showed positive bands implying that this accession might harbor Bph10 and Bph18(t) genes. Three aromatic accessions, Domsiah, Khao Dawk Mali 105 and 415XIr352 showed gene pyramiding of Bph1 and Bph13(t). Two indica aromatic rice, Ds 20 and Basmati 9-93, possessed at least two BPH resistance genes, Bph1, Bph18(t) and Bph13(t), Bph18(t), respectively. These results indicates that aromatic rice germplasm have narrow diversities of BPR 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.

Screening of the Dominant Rice Blast Resistance Genes with PCR-based SNP and CAPS Marker in Aromatic Rice Germplasm

  • Kim, Jeong-Soon;Ahn, Sang-Nag;Hong, Sung-Jun;Kwon, Jin-Hyeuk;Kim, Yeong-Ki;Jee, Hyeong-Jin;Shim, Chang-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.4
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    • pp.329-341
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    • 2011
  • The objective of this study was to determine the genetic diversities of major rice blast resistance genes among 84 accessions of aromatic rice germplasm. Eighty four accessions were characterized by a dominant 11 set of PCR-based SNP and CAPS marker, which showed the broad spectrum resistance and closest linkage to seven major rice blast resistance (R) genes, Pia, Pib, Pii, Pi5 (Pi3), Pita (Pita-2), and Pi9 (t). The allele specific PCR markers assay genotype of SCAR and STS markers was applied to estimate the presence or absence of PCR amplicons detected with a pair of PCR markers. One indica accession, Basmati (IT211194), showed the positive amplicons of five major rice blast resistance genes, Pia, Pi5 (Pi3), Pib, Pi-ta (Pi-ta2), and Pik-5 (Pish). Among 48 accessions of the PCR amplicons detected with yca72 marker, only five accessions were identified to Pia gene on chromosome 11. The Pib gene was estimated with the NSb marker and was detected in 65 of 84 accessions. This study showed that nine of 84 accessions contained the Pii gene and owned Pi5 (Pi3) in 42 of 84 accessions by JJ817 and JJ113-T markers, which is coclosest with Pii on chromosome 9. Only six accessions were detected two alleles of the Pita or Pita-2 genes. Three of accessions were identified as the Pi9 (t) gene locus.