• Title/Summary/Keyword: Polymorphic markers

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In Vitro Selection and Characterizations of Gamma Radiation-Induced Salt Tolerant Lines in Rice (방사선을 이용한 내염성 계통의 기내선발 및 특징)

  • Lee, In-Sok;Kim, Dong-Sub;Hyun, Do-Yoon;Lim, Yong-Pyo;Lee, Young-Il
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.247-252
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    • 2002
  • The combination of radiation technique with an in vitro culture system was initiated to develop salt tolerant rice. We established an in vitro culture system to select tolerant lines against salt stress. NaCl tolerant cell lines were selected from the callus irradiated with gamma ray on N$_{6}$ medium with 1.5% NaCl and 2 mg/L 2,4-D. Regenerants (M$_1$) were obtained from the tolerant callus which was cultured for 30 days auxin-free medium. The M$_2$seeds were harvested from M$_1$plants on an individual plant basis. Thirty seedlings from each 450 M$_2$lines were transplanted in a field and total 5,000 M$_3$lines were harvested with an average 90 percent of fertile grain. M$_3$lines were utilized for selection of salt tolerance. Salinity-tolerant lines (225) were selected among 5,000 M$_3$lines. Of the 225 lines tested, the morphological traits of two lines (120-10 and -11) were far superior to control (Donagjinbyeo) in agromomic traits such as plant height, root length and no. of roots. Control and tolerant lines were analyzed by RAPD markers. Three polymorphic bands were presented in only tolerant lines, demonstrating a genetic difference between control and the tolerant lines. Such tolerant lines could be used as genetic resources to improve salt tolerance.e.

Assessment of genetic diversity and population structure of commercial button mushroom (Agaricus bisporus) strains in Korea (한국의 상업적 양송이 균주의 유전적 다양성 및 집단 구조)

  • Lee, Hwa-Yong;An, Hye-jin;Oh, Youn-Lee;Jang, Kab-Yeul;Kong, Won-Sik;Ryu, Ho-jin;Chung, Jong-Wook
    • Journal of Mushroom
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    • v.17 no.4
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    • pp.171-178
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    • 2019
  • Agaricus bisporus is a functional food and among the most widely cultivated mushrooms in the world. In this study, we analyzed the genetic diversity and population structure of 23 Korean and 42 foreign A. bisporus cultivars using SSR (Simple sequence repeat) markers. Genetic diversity of A. bisporus cultivars was as follows: number of alleles was approximately 13; observed and expected heterozygosity were approximately 0.59 and 0.74, respectively; and polymorphic information content value was about 0.71. A. bisporus cultivars were divided into three groups using distance-based analysis. Genetic diversity of Group 2, which consisted of cultivars from various countries, was high. In addition, model-based subpopulations were divided into two, and the genetic diversity of Pop2 (Population 2), which had many cultivars, was high. The results of this study could be used in a breeding program for A. bisporus, such as the development of new genetic resources and securing diversity.

Construction of a Microsatellite DNA Profile Database for Pear Cultivars and Germplasm (배 품종 및 유전자원에 대한 Microsatellite DNA 프로파일 데이터베이스 구축)

  • Hong, Jee-Hwa;Shim, Eun-Jo;Kwon, Yong-Sham
    • Horticultural Science & Technology
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    • v.35 no.1
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    • pp.98-107
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    • 2017
  • A DNA profile database was constructed to investigate the genetic relatedness of 72 germplasm samples of Pyrus and related cultivars using microsatellite markers. Three P. pyrifolia, four P. commus, and one P. betulifolia cultivars with different morphological traits were screened using 387 pairs of microsatellite primers. A core set of 11 primer pairs was selected to obtain 133 polymorphic amplified fragments meeting three criteria: high polymorphism information contents (PIC), high repeatability, and distinct allele patterns. The number of alleles per locus ranged between 4 and 22. Average PIC was 0.743 (range: 0.557 - 0.879). Cluster analysis using the unweighted pair - group method with arithmetical average (UPGMA) separated the 72 pear cultivars and germplasm samples into four major groups: Chinese, European pears, and a cluster of 55 Asian pears that could be reclassify into two subcluster, I - $1^{st}$ and II - $2^{nd}$, according to pedigree information. Almost all of the cultivars were discriminated by 11 microsatellite marker genotypes. The microsatellite DNA profile database may be utilized as tool to verify distinctness, uniformity, and stability between candidate cultivar, and to verify in the distinctness of existing cultivars.

Development of Sequence Characterized Amplified Regions (SCAR) Showing for Cheju Native Horse (품종 특이성을 이용한 제주마 판별 표지인자 재발)

  • Cho Byung Wook
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.474-478
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    • 2005
  • This study was conducted to analyze genetic characteristics and to develop the specific marker for Cheju native horse (Coo) at the level of sequence characterized amplified regions (SCARs). We collected blood samples from Cheju native horse and Thoroughbred horse (Th) and obtained genomic DNA from the blood of 50 individuals randomly selected within the breeds. Seven hundred primers were chosen randomly and were used to examin the polymorphism and 40 kinds of primers showed polymorphic RAPD band patterns between two breeds. Thirty primers of them showed horse specific bands. With the primer MG 30, amplified band of 2.0 kb showed the specificity to Cheju native horse (Cnh). Additionally MG 53 detected the thoroughbred horse (Th) specific markers at size of 2.3 kb. As the next, 2.3 kb band from MG 53 was checked with the all individuals from all the breeds of this study, and it maintained the reproducible breed specificity to thoroughbred horse (Th). With this results, 2.3 kb band was cloned into plasmid vector and sequenced bidirectionally from both ends of the cloned fragment. With the obtained sequences 10 nucleotide extended primers including the original arbitray primer were designed as a SCARs primer. Finally, the primer with extended sequence showed the reproducible breed differentiation pattern and it was possible to identify Cheju native horse (Cnh) from other breeds. The SCARs marker 2.3 kb from MG 53 could be used to identify Cheju native horse (Cnh) for not only registration but also horse breeding programe.

Luteinizing hormone beta gene polymorphism and its effect on semen quality traits and luteinizing hormone concentrations in Murrah buffalo bulls

  • Reen, Jagish Kour;Kerekoppa, Ramesha;Deginal, Revanasiddu;Ahirwar, Maneesh Kumar;Kannegundla, Uday;Chandra, Satish;Palat, Divya;Das, Dayal Nitai;Kataktalware, Mukund Amritrao;Jeyakumar, Sakthivel;Isloor, Shri krishna
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1119-1126
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    • 2018
  • Objective: Present investigation was aimed to study the Single Nucleotide Variants of the luteinizing hormone beta ($LH{\beta}$) gene and to analyze their association with the semen quality (fresh and post-thawed frozen semen) and luteinizing hormone (LH) concentrations in Murrah buffalo bulls. Methods: Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) and Sanger sequencing method is used to study genetic variability in $LH{\beta}$ gene. LH assay was carried out using enzyme-linked immunosorbent assay method. A fixed general linear model was used to analyze association of single nucleotide polymorphism (SNP) of $LH{\beta}$ gene with semen quality in 109 and LH concentrations in 80 Murrah bulls. Results: $LH{\beta}$ gene was found to be polymorphic. Total six SNPs were identified in $LH{\beta}$ gene g C356090A, g C356113T, g A356701G, g G355869A, g G356330C, and g G356606T. Single Stranded Conformational Polymorphism variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on sperm concentration (million/mL), percent mass motility, acrosome integrity and membrane integrity in fresh and frozen semen whereas significant (p<0.05) effect was observed on percent live spermatozoa. SSCP variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on luteinizing hormone concentrations too. Conclusion: The observed association between SSCP variants of $LH{\beta}$ gene with semen quality parameters and LH concentrations indicated the possibilities of using $LH{\beta}$ as a candidate gene for identification of markers for semen quality traits and LH concentrations in Murrah buffaloes.

Development of a CAPS Marker Derived from the Pg-Actin Gene Sequences and RAPD Markers in Platycodon grandiflorum (도라지에서의 RAPD 마커 분석과 Actin 유전자 염기서열에서 유래한 CAPS 분자표지 개발)

  • Kim, Munhwi;Jeong, Eunah;Jeong, Jeongsu;Kwon, Soontae;Jeon, Ikjo;Jeong, Jeong Hag;Lee, Je Min;Yeam, Inhwa
    • Korean Journal of Plant Resources
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    • v.28 no.5
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    • pp.648-655
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    • 2015
  • Balloon flower (Platycodon grandiflorum A. DC.) is a perennial plant of mainly Campanulaceae family, which have been widely used as a food ingredient and herbal medicine in East Asia. Although demands on related products and yearly cultivation area for balloon flower are increasing, diverse fundamental technologies and molecular breeding studies are not very well supported in Platycodons. In this study, 30 random amplification of polymorphic DNA (RAPD) primers were test in an attempt to explore genetic diversities. In addition, sequences information of the actin gene, a well conserved gene encoding a globular protein that forms microfilaments, was retrieved and analyzed. Two actin homologs were recovered; 3.4 kb fragment is a Pg-actin and 1.4 kb fragment is a Pg-actin homolog with 28.6% similarity. We have confirmed that the Pg-actin gene is configured into 4 exons and 3 introns. A single nucleotide polymorphism (SNP), G↔A, was detected on the intron 3, which served as a target for the CAPS marker development. The marker Pg-Actin-Int3 was applied to 32 balloon flower accessions. Balloon flower DNA sequence information generated in this study is expected to contribute to the analysis and molecular breeding and genetic diversity analysis of balloon flowers.

Development and Characterization of Chloroplast Simple Sequence Repeat markers in Pinus koraiensis (잣나무 엽록체 Simple Sequence Repeat 표지자 개발 및 특성 분석)

  • Lee, Jei-Wan;Baek, Seung-Hoon;Hong, Kyung-Nak;Hong, Yong-Pyo;Lee, Seok-Woo;Ahn, Ji-Young
    • Journal of Korean Society of Forest Science
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    • v.104 no.4
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    • pp.549-557
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    • 2015
  • Novel cpSSR primers were developed based on the sequence information of the Pinus koraiensis chloroplast genome. A total of 30 cpSSR loci were detected in the chloroplast genome, and a total of 30 primer sets flanking those loci were designed. All primer sets were successfully amplified for chloroplast DNA in P. koraiensis. The cross-species transferability of the 30 primer sets was considerably high in P. pumila (100%) and P. paviflora (97%) belonging to the same Subgenus (Strobus) of P. koraiensis. Meanwhile, the transferability was relatively low (73%) in P. densiflora and P. sylvestris belonging to Subgenus Pinus. A total of 13 cpSSR loci out of the 30 loci were polymorphic in the Mt. Jumbong population of P. koraiensis. The mean of haploid diversity(H) was 0.512. The number of haplotypes(N) and the haplotype diversity($H_e$) were 25 and 0.992, respectively. Of the 25 haplotypes, 22 were unique in the analyzed population. The unique haplotypes differentiated 22 individuals (79%) from the total of 28 individuals. In conclusion, the novel cpSSR primers developed in this study would be applicable to other Pinus species, especially the subgenus Strobus, and provide a high level of polymorphism for the study of genetic variation of P. koraiensis.

Classification of Microsatellite Alterations Detected in Endoscopic Biopsy Specimens of Gastric Cancers (단순반복염기서열의 변이 형태에 따른 위암 내시경 조직의 유전자형 분류)

  • Choi Young Deok;Choi Sang Wook;Jeon Eun Jeong;Jeong Jeong Jo;Min Ki Ouk;Lee Kang Hoon;Lee Sung;Rhyu Mun Gan
    • Journal of Gastric Cancer
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    • v.4 no.2
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    • pp.109-120
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    • 2004
  • Purpose: Individual gastric cancers demonstrate complicated genetic alterations. The PCR-based analysis of polymorphic microsatellite sequences on cancer-related chromosomes has been used to detect chromosomal loss and microsatellite instability. For the purpose of preoperative usage, we analyzed the correspondance rate of the microsatellite genotype between endoscopic biopsy and surgical specimens. Materials and Methods: Seventy-three pairs of biopsy and surgical specimens were examined for loss of heterozygosity and microsatellite instability by using 40 microsatellite markers on eight chromosomes. Microsatellite alterations in tumor DNAs were classified into a high-risk group (baselinelevel loss of heterozygosity: 1 chromosomal loss in diffuse type and high-level loss of heterozygosity: 4 or more chromosomal losses) and a low-risk group (microsatellite instability and low-level loss of heterozygosity: 2 or 3 chromosomal losses in diffuse type or $1\∼3$ chromosomal losses in intestinal type) based on the extent of chromosomal loss and microsatellite instability. Results: The chromosomal losses of the biopsy and the surgical specimens were found to be different in 21 of the 73 cases, 19 cases of which were categorized into a genotype group of similar extent. In 100 surgical specimens, the high-risk genotype group showed a high incidence of nodal involvement (19 of 23 cases: $\leq$5 cm; 23 of 24 cases: >5 cm) irrespective of tumor size while the incidence of nodal involvement for the low-risk genotype group depended on tumor size (5 of 26 cases: $\leq$5 cm; 18 of 27 cases: >5 cm). Extraserosal invasion was more frequent in large-sized tumor in both the high-risk genotype group ($\leq$5 cm: 12 of 23 cases; >5 cm: 23 of 24 cases) and the low-risk genotype group ($\leq$5 cm: 7 of 26 cases; >5 cm: 16 of 27 cases). The preoperative prediction of tumor invasion and nodal involvement based on tumor size and genotype corresponded closely to the pathologic tumor stage (ROC area >0.7). Conclusion: An endoscopic biopsy specimen of gastric cancer can be used to make a preoperative genetic diagnosis that accurately reflect the genotype of the corresponding surgical specimen.

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Relationship Between Heterosis and Genetic Divergence in Tongil-type Rice(Oryza sativa L.) (벼에서 RAPDs 분석에 의한 양친의 유전적 유사성과 잡종강세의 상관)

  • Ahn, Sang-Nag;Kim, Yun-Sun
    • Korean Journal of Agricultural Science
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    • v.28 no.1
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    • pp.1-7
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    • 2001
  • In this study we evaluated genetic divergence among six Tongil-type rice varieties and assessed the relationship between genetic distance and hybrid performance in all possible non-reciprocal crosses between them. The 15 F1 hybrids along with the six parents were evaluated for eight traits of agronomic importance including yield in a replicated field trial. The six parents were examined for DNA polymorphism using 120 random decamer oligonucleotide primers. A total of 168 polymorphic variants were generated and based on the polymorphism data, genetic distances (GDs) ranged from 0.157 to 0.383. Heterosis was observed in hybrids for most of the traits, and yield exhibited the highest heterosis among the eight traits examined. The correlation values of GDs with F1 performance were not significant. Also the correlations of GDs with midparent and better-parent heterosis were not significant except for spikelets per panicle. Our results have indicated that GDs based on the RAPDs markers may not be useful for predicting heterotic combinations in Tongil-type rice and are supportive of the idea that the level of correlations between hybrid performance and genetic divergence is dependent on the germlplasm employed.

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Needle Characteristics and Genetic Variation of Pinus pumila Natural Population in Mt. Seorak (설악산 눈잣나무 천연집단의 침엽특성과 유전변이)

  • Song, Jeong-Ho;Lim, Hyo-In;Jang, Kyung-Hwan
    • Korean Journal of Plant Resources
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    • v.25 no.5
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    • pp.517-522
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    • 2012
  • This study was conducted to investigate the morphological and anatomical characteristics of needle and the genetic diversity of Pinus pumila Regel which is a unique and the southern peripheral population in South Korea. ANOVA test showed that there were significant differences among individuals within population in all 8 needle characteristics. Average characteristics of 66 individuals were 53.59 mm in needle length, 0.78 mm in needle width, 68.98 in needle index, 0.65 mm in needle thickness, 4.56 ea. in maximum stomata row, 3.80 ea. in minimum stomata row, 8.36 ea. in total stomata row and 1.71 ea. in resin canals, respectively. Resin canal per needle of this species ranged from one to three, depending on external type. Especially, arrangement types were 69.47% in two resin canals and 30.45% in a single resin canal. A total of 78 bands was generated from 9 selected I-SSR primers. The estimates of genetic variation were 61.5% in proportion of polymorphic bands (P), 1.698 in effective number of alleles ($A_e$), 0.388 in expected heterozygosity ($H_e$) and 0.567 in Shannon's information index (S.I.), respectively.