• Title/Summary/Keyword: SCAR markers

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Interseeding Substitution Ratio Assessment by Using of Molecular Tool (분자생물학적 기법을 이용한 Interseeding률 평가)

  • Jung, Seung-Ho;Lim, Deuk-Jong;Jang, Duk-Hwan
    • Asian Journal of Turfgrass Science
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    • v.24 no.2
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    • pp.131-137
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    • 2010
  • Recently, many domestic golf course have showed interest in introducing new cultivars into existing putting greens without reconstruction. But, any report and survey method for interseeding substitute ratio haven't been published. Therefore, this study was conducted to evaluate the effectiveness for interseeding by investigating how many interseeding plants were introduced into existing putting green by using molecular biologic tool. The Woo Jung Hills Country Club was one time introduced penn A4 into penncross putting green. The Lake Side Country Club was three times introduced CY-2 into penncross putting green. The interseeding substitute ratio using SCAR markers was 20% in the Woo Jung Hills Country Club and 35.5% in Lake Side Country Club. The interseeding putting green in Lake Side Country Club was investigated to three site, strong pressure, middle pressure and weak pressure. These substitute ratios were strong pressure 27.1%, middle pressure 37.3%, and weak pressure 47.2%. The data indicated that an interseeding substitute ratio was more increased three times than one time to introducing new cultivar into existing putting green. A long-term program should be made in order to exchange existing cultivar in putting green without reconstruction.

Development of an ISSR-Derived SCAR Marker in Korean Ginseng Cultivars (Panax ginseng C. A. Meyer)

  • Lee, Jei-Wan;Kim, Young-Chang;Jo, Ick-Hyun;Seo, A-Yeon;Lee, Jeong-Hoon;Kim, Ok-Tae;Hyun, Dong-Yun;Cha, Seon-Woo;Bang, Kyong-Hwan;Cho, Joon-Hyeong
    • Journal of Ginseng Research
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    • v.35 no.1
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    • pp.52-59
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    • 2011
  • Recently, new ginseng cultivars having superior agricultural traits have been developed in Korea. For newly developed plant cultivars, the identification of distinctiveness is very important factors not only in plant cultivar management but also in breeding programs. Thus, eighty-five inter simple sequence repeat (ISSR) primers were applied to detect polymorphisms among six major Korean ginseng cultivars and two foreign ginsengs. A total of 197 polymorphic bands with an average 5.8 polymorphic bands and 2.9 banding patterns per assay unit across six Korean ginseng cultivars and foreign ginsengs from 236 amplified ISSR loci with an average 6.9 loci per assay unit were generated by 34 out of 85 ISSR primers. Three species of Panax ginseng including the Korean ginseng cultivars, P. quinquefolius, and P. notoginseng, could be readily discriminated using most tested primers. UBC-821, UBC-868, and UBC-878 generated polymorphic bands among the six Korean ginseng cultivars, and could distinguish them from foreign ginsengs. Sequence characterized amplified region (SCAR) marker system was introduced in order to increase the reproducibility of the polymorphism. One SCAR marker, PgI821C650, was successfully converted from the randomly amplified polymorphism by UBC-821. It showed the expected dominant polymorphism among ginseng samples. In addition, the specific polymorphism for Sunwon was generated by treating Taq I restriction enzyme to polymerase chain reaction products of PgI821C650. These results will serve as useful DNA markers for identification of Korean ginseng, especially Sunwon cultivar, seed management, and molecular breeding program supplemented with marker-assisted selection.

The Construction of a Chinese Cabbage Marker-assisted Backcrossing System Using High-throughput Genotyping Technology

  • Kim, Jinhee;Kim, Do-Sun;Lee, Eun Su;Ahn, Yul-Kyun;Chae, Won Byoung;Lee, Soo-Seong
    • Horticultural Science & Technology
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    • v.35 no.2
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    • pp.232-242
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    • 2017
  • The goal of marker-assisted backcrossing (MAB) is to significantly reduce the number of breeding generations required by using genome-based molecular markers to select for a particular trait; however, MAB systems have only been developed for a few vegetable crops to date. Among the types of molecular markers, SNPs (single-nucleotide polymorphisms) are primarily used in the analysis of genetic diversity due to their abundance throughout most genomes. To develop a MAB system in Chinese cabbage, a high-throughput (HT) marker system was used, based on a previously developed set of 468 SNP probes (BraMAB1, Brassica Marker Assisted Backcrossing SNP 1). We selected a broad-spectrum TuMV (Turnip mosaic virus) resistance (trs) Chinese cabbage line (SB22) as a donor plant, constructing a $BC_1F_1$ population by crossing it with the TuMV-susceptible 12mo-682-1 elite line. Foreground selection was performed using the previously developed trsSCAR marker. Background selection was performed using 119 SNP markers that showed clear polymorphism between donor and recipient plants. The background genome recovery rate (% recurrent parent genome recovery; RPG) was good, with three of 75 $BC_1F_1$ plants showing a high RPG rate of over 80%. The background genotyping result and the phenotypic similarity between the recurrent parent and $BC_1F_1$ showed a correlation. The plant with the highest RPG recovery rate was backcrossed to construct the $BC_2F_1$ population. Foreground selection and background selection were performed using 169 $BC_2F_1$ plants. This study shows that, using MAB, we can recover over 90% of the background genome in only two generations, highlighting the MAB system using HT markers as a highly efficient Brassica rapa backcross breeding system. This is the first report of the application of a SNP marker set to the background selection of Chinese cabbage using HT SNP genotyping technology.

Development of PCR-Based Sequence Characterized DNA Markers for the Identification and Detection, Genetic Diversity of Didymella bryoniae with Random Amplified polymorphic DNA(RAPD)

  • Kyo, Seo-Il;Shim, Chang-Ki;Kim, Dong-Kil;Baep, Dong-Won;Lee, Seon-Chul;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.130-130
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    • 2003
  • Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

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PCR-based markers to select plastid genotypes of Solanum acaule (Solanum acaule 색소체 유전자형 선발을 위한 특이적 분자마커 개발)

  • Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.178-186
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    • 2022
  • The tetraploid Solanum acaule is a wild potato species from Bolivia widely used for potato breeding because of its diverse attractive traits, including resistance to frost, late blight, potato virus X, potato virus Y, potato leafroll virus, potato spindle tuber viroid, and cyst nematode. However, the introgression of useful traits into cultivated potatoes via crossing has been limited by differences in endosperm balance number between species. Somatic fusion could be used to overcome sexual reproduction barriers and the development of molecular markers is essential to select proper fusion products. The chloroplast genome of S. acaule was sequenced using next-generation sequencing technology and specific markers for S. acaule were developed by comparing the obtained sequence with those of seven other Solanum species. The total length of the chloroplast genome is 155,570 bp, and 158 genes were annotated. Structure and gene content were very similar to other Solanum species and maximum likelihood phylogenetic analysis with 12 other species belonging to the Solanaceae family revealed that S. acaule is very closely related to other Solanum species. Sequence alignment with the chloroplast genome of seven other Solanum species revealed four InDels and 79 SNPs specific to S. acaule. Based on these InDel and SNP regions, one SCAR marker and one CAPS marker were developed to discriminate S. acaule from other Solanum species. These results will aid in exploring evolutionary aspects of Solanum species and accelerating potato breeding using S. acaule.

S-haplotypes and Genetic Diversity in 'Danji' Radish (Raphanus sativus L. var. hortensis)

  • Ahn, Yulkyun;Kim, Hyukjun;Han, Dongyeop;Park, Younghoon
    • Horticultural Science & Technology
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    • v.32 no.2
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    • pp.210-216
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    • 2014
  • The distribution of S-haplotypes and genetic relationships were evaluated for 47 accessions of 'Danji' radish (Raphanus sativus L. var. hortensis Baker f. gigantissimus Makino) originating from Jeju Island in South Korea. A total of 22 S-haplotype-specific SCAR markers for the S locus glycoprotein (SLG) and S receptor kinase (SRK) loci were tested, and six primer sets amplified locus-specific PCR fragments from at least one 'Danji' radish accession. S5 and S21 alleles atthe SLG locus were the most frequently distributed, and detected from 87.5% and 64.6% of the accessions, respectively. The frequency of the class-II haplotype at the SLG locus was 75%, more frequent than the class-I haplotype. The S23 allele at the SRK locus was detected from 7 accessions. Grouping of the accessions based on S-allele composition revealed three major groups, while 8 accessions showed a unique allelic composition. The genetic diversity of 47 'Danji' radishes and 1 'Gwandong' radish were also evaluated with 38 RAPD primers. A total of 312 bands were scored, and showed that 138 bands (44.2%) were monomorphic among the accessions, whereas 174 (55.8%) bands were polymorphic. Polymorphism rates ranged from 0.2 to 1.0, indicating significant variations in detecting polymorphism across RAPD primers. The genetic similarity coefficients among all pairs of the 48accessions varied from 0.62 to 0.93, and 42% of the comparisons exhibited values higher than 0.85. All the cultivars could be distinguished based on the DNA fingerprints revealed by RAPD. The comparisons between the dendrograms based on S-haplotypes and RAPDs indicate an unrelated and sporadic distribution for several accessions; however, there was a tendency for accessions with the same S-allelic composition to group into the same cluster.

Resistance Screening to Pepper mild mottle virus Pathotypes in Paprika Cultivars (고추약한모틀바이러스 병원형에 대한 파프리카 품종의 저항성 스크리닝)

  • Choi, Gug-Seoun;Choi, Seung-Kook;Cho, In-Sook;Kwon, Sun-Jung
    • Research in Plant Disease
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    • v.20 no.4
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    • pp.299-302
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    • 2014
  • The Paprika plant infected with Pepper mild mottle virus (PMMoV) do not produce commercial fruit as causing necrotic spots symptom on the fruit. Ten cultivars of paprika were analyzed to select the resistance cultivars against PMMoV pathotypes, $P_{1.2}$ and $P_{1.2.3}$, using bioassay and genetic markers. $L^1$, $L^3$, and $L^4$ genotypes expressing resistance to the pathotypes existed in those cultivars but $L^2$ genotype did not. $L^4L^4$ in cvs. Easy and Magnifico, $L^4L^3$ in cvs. Scirocco and Orange glory F1, $L^4L^1$ in cv. Special F1, $L^3L^3$ in cvs. Fiesta, Piero and Derby, and $L^3L^1$ in Cupra and Mazzona F1 were identified with SCAR and CAPS markers. The resistant cvs. to the 2 pathotypes were Magnipico, Easy, Scirocco F1, Orange glory and Special F1 and the susceptible cvs. were Fiesta, Piero, Derby, Cupra and Mazzona F1. The susceptible cvs. of the absence of $L^4$ genotype showed systemic infection when inoculated with PMMoV-$P_{1.2.3}$. However, those cvs. despite the presence of $L^3$ genotype showed vein necrosis on the inoculated leaf and hypersensitive necrosis symptom on the upper parts when inoculated with PMMoV-$P_{1.2}$.

The Clinical Comparison between Monomicrobial and Polymicrobial Urinary Infection in Febrile Pediatric Acute Pyelonephritis (발열성 소아 신우 신염에서 단일 세균 감염과 혼합 세균 감염의 임상적 비교)

  • Lee, In Hak;Nam, Seong Woo;Seo, Hyeon Seok;Yim, Hyung Eun;Yoo, Kee Hwan;Hong, Young Sook;Lee, Joo Won
    • Childhood Kidney Diseases
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    • v.16 no.2
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    • pp.102-108
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    • 2012
  • Purpose: We investigated the clinical presentation of febrile pediatric patients with acute pyelonephritis (APN) with a mixed urine culture from an aseptic urine sample, and compared with that of those with a single culture. Methods: We retrospectively reviewed the medical charts of 95 patients diagnosed as APN with fever between January 2008 and October 2010 at Korea University Medical Center. We classified the patients with APN into two groups with a positive single culture (S group) and a positive mixed culture (M group) from an aseptic urine sample of suprapubic bladder aspiration or urethral catheterization and compared the fever duration, laboratory markers such as serum white blood cell (WBC) counts and C-reactive protein (CRP) values in peripheral blood, and the presence of hydronephrosis, renal scar and vesicoureteral reflux (VUR) between the two groups (If presence of hydronephrosis, scar and VUR=1 and no=0). Results: Total pediatric patients with febrile APN were 95 patients, a positive S group was 89 patients and a positive M group was 6 patients. Fever duration (S vs. M, $4.7{\pm}3.1$ vs. $6{\pm}5.7$ days), serum WBC (S vs. M, $18,630{\pm}6,483$ vs. $20,153{\pm}7,660/{\mu}L$) and CRP (S vs. M, $100.6{\pm}2.46$ vs. $81.1{\pm}0.09\;mg/L$) values, and the presence of hydronephrosis, renal scar and VUR were not different between the two groups. Conclusion: Our data shows that there were no specific differences of clinical manifestation between a positive single urine culture and a positive mixed urine culture in pediatric APN. A mixed urine culture from an aseptic urine sample should be interpreted cautiously.

Recurrent parent genome (RPG) recovery analysis in a marker-assisted backcross breeding based on the genotyping-by-sequencing in tomato (Solanum lycopersicum L.) (토마토 MABC 육종에서 GBS(genotyping-by-sequencing)에 의한 RPG(recurrent parent genome) 회복률 분석)

  • Kim, Jong Hee;Jung, Yu Jin;Seo, Hoon Kyo;Kim, Myong-Kwon;Nou, Ill-Sup;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.165-171
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    • 2019
  • Marker-assisted backcrossing (MABC) is useful for selecting an offspring with a highly recovered genetic background for a recurrent parent at early generation to various crops. Moreover, marker-assisted backcrossing (MABC) along with marker-assisted selection (MAS) contributes immensely to overcome the main limitation of the conventional breeding and it accelerates recurrent parent genome (RPG) recovery. In this study, we were employed to incorporate rin gene(s) from the donor parent T13-1084, into the genetic background of HK13-1151, a popular high-yielding tomato elite inbred line that is a pink color fruit, in order to develop a rin HK13-1084 improved line. The recurrent parent genome recovery was analyzed in early generations of backcrossing using SNP markers obtained from genotyping-by-sequencing analysis. From the $BC_1F_1$ and $BC_2F_1$ plants, 3,086 and 4868 polymorphic SNP markers were obtained via GBS analysis, respectively. These markers were present in all twelve chromosomes. The background analysis revealed that the extent of RPG recovery ranged from 56.7% to 84.5% and from 87.8% to 97.8% in $BC_1F_1$ and $BC_2F_1$ generations, respectively. In this study, No 5-1 with 97.8% RPG recovery rate among $BC_2F_1$ plants was similar to HK13-1151 strain in the fruit shape. Therefore, the selected plants were fixed in $BC_2F_2$ generation through selfing. MAS allowed identification of the plants that are more similar to the recurrent parent for the loci evaluated in the backcross generations. MABC can greatly reduce breeding time as compared to the conventional backcross breeding. For instance, MABC approach greatly shortened breeding time in tomato.

The Study of the Availability of Acute Reactive Markers in Children with Upper Urinary Tract Infection (소아의 상부요로감염에서 급성기 반응지표의 유용성에 관한 연구)

  • Lee, Hye Yeong;Lee, Baeck Hee
    • Pediatric Infection and Vaccine
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    • v.5 no.2
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    • pp.221-229
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    • 1998
  • Purpose : The symptom of urinary tract infection in children is non-specific and systemic compared to that of adults. So the diagnosis of UTI in children can be delayed. If UTI in children is not appropriately managed, it may he resulted in renal failure, hypertension, growth failure in adolescence because there are already documented renal scar or urinary tract abnormality in infection period. Especially upper UTI that involve renal parenchyme may be result in fatal complication. The purpose of this study is analyzing the relationship between acute reactive marker and $^{99m}TC$-DMSA renal scan in upper urinary tract infection. Methods : This study included 56 children admitted at Dankook University Hospital Pediatric Department in Jan. 1995~May. 1998. We analyzed quantatively the results of acute reactive marker(CRP, ESR, WBC), pyuria, fever and compared to those of sonographically find ing and $^{99m}TC$-DMSA renal scan. Comparison between groups were performed by the chi-square (x2) test and a p value of less than 0.05 was considered statistically significant. Results : 1) The number of boys less than 1 year of age was larger than that of girls. But the number of boys more than 1 year of age was reversed. 2) The higher me level of reactive marker (CRP, WBC), the more the probability of upper UTI. 3) The higher fever, the more the probability of upper UTI. 4) The more pyuria, the more probability of upper UTI. 5) The more higher the grade of vesicoureteral reflux, the more probability of upper UTI. 6) $^{99m}TC$-DMSA renal scan is more sensitive and more specific diagnostic tool than renal sonogram. Conclusion : The appearance of an abnormal $^{99m}TC$-DMSA renal scan is correlated with acute reactive marker (CRP, ESR, WBC), fever, pyuria. $^{99m}TC$-DMSA renal scan can be a good valuable predictor tool in upper UTI. So we can start early treatment and decrease the incidence of complication of upper urinary tract by above indicators before knowing the result of urine culture. And we can follow up the patients in more good relationships with their parents by telling them the duration of treatment and follow-up plan.

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