• Title/Summary/Keyword: Polymorphic markers

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Development of Functional Markers for Detection of Inactive DFR-A Alleles Responsible for Failure of Anthocyanin Production in Onions (Allium cepa L.)

  • Park, Jaehyuk;Cho, Dong Youn;Moon, Jin Seong;Yoon, Moo-Kyoung;Kim, Sunggil
    • Horticultural Science & Technology
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    • v.31 no.1
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    • pp.72-79
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    • 2013
  • Inactivation of the gene coding for dihydroflavonol 4-reductase (DFR) is responsible for the color difference between red and yellow onions (Allium cepa L.). Two inactive DFR-A alleles, DFR-$A^{PS}$ and DFR-$A^{DEL}$, were identified in our previous study. A functional marker was developed on the basis of the premature stop codon that inactivated the DFR-$A^{PS}$ allele. A derived cleaved amplified polymorphic sequences (dCAPS) primer was designed to detect the single nucleotide polymorphism, an A/T transition, which produced the premature stop codon. Digested PCR products clearly distinguished the homozygous and heterozygous red $F_2$ individuals. Meanwhile, to develop a molecular marker for detection of the DFR-$A^{DEL}$ allele in which entire DFR-A gene was deleted, genome walking was performed and approximately 3 kb 5' and 3' flanking sequences of the DFR-$A^R$ coding region were obtained. PCR amplification using multiple primers binding to the extended flanking regions showed that more of the extended region of the DFR-A gene was deleted in the DFR-$A^{DEL}$ allele. A dominant simple PCR marker was developed to identify the DFR-$A^{DEL}$ allele using the dissimilar 3' flanking sequences of the DFR-A gene and homologous DFR-B pseudogene. Distribution of the DFR-$A^{PS}$ and DFR-$A^{DEL}$ alleles in yellow onion cultivars bred in Korea and Japan was surveyed using molecular makers developed in this study. Results showed predominant existence of the DFR-$A^{PS}$ allele in yellow onion cultivars.

Genetic Relationship Analysis of genus Nelumbo Accessions Based on Inter-Simple Sequence Repeats (ISSR) (ISSR 표지에 의한 연속 (Nelumbo)의 유연관계 분석)

  • Ryu, Jai-Hyunk;Choi, Gab-Lim;Lyu, Jae-Il;Lee, Sheong-Chun;Chun, Jong-Un;Shin, Dong-Young;Bae, Chang-Hyu
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.2
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    • pp.86-92
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    • 2010
  • The polymorphism and the genetic relationships among 32 genetic resources of genus Nelumbo from Korea, Japan, China, USA, India, Thailand and Gabong were thoroughly investigated and extensively examined using ISSR markers. Out of 103 loci detected overall, 94 were identified to be polymorphic with a rate of 91.2%. The genetic similarity matrix revealed a wide range of variability among the 32 accessions, spanning from 0.227 to 0.833. The study findings indicate that the Nelumbo accessions have a high genetic diversity, and accordingly carry a germplasm qualifying as good genetic resources for cross breeding. According to the clustering analysis, different subspecies, N. nucifera and N. lutea, were divided into independent groups and all of the N. nucifera accessions could be classified into five categories. Compared to RAPD analysis, ISSR method showed a clearer picture of polymorphism among the accessions and exhibited a definite distinction even among the subspecies. In this respect, ISSR analysis is considered to be more effective in differentiating the accessions and subspecies of the genus Nelumbo than RAPD test.

Ten new microsatellite markers in cutlassfish Trichiurus lepturus derived from an enriched genomic library

  • An, Hye-Suck;Lee, Jeong-Ho;Noh, Jae-Koo;Kim, Hyun-Chul;Park, Chul-Ji;Min, Byung-Hwa;Myeong, Jeong-In
    • Animal cells and systems
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    • v.14 no.3
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    • pp.169-174
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    • 2010
  • Cutlassfish (Trichiurus lepturus Linnaeus 1758) is a commercially important fish in Korea. In recent years, the catch of cutlassfish in the coastal waters of Korea has significantly declined. Its genetic characterization has been little studied. To assist conservation and management efforts, we isolated and characterized 10 microsatellite loci using an enrichment method based on magnetic/biotin capture of microsatellite sequences from a size-selected genomic library. To characterize each locus, 30 individuals from a natural T. lepturus population in the coastal waters of Jeju Island, Korea, were genotyped. All loci except two, KTh9B and KTh22A, were polymorphic, with an average of 14.3 alleles per locus (range, 10 22). The mean observed and expected heterozygosities were 0.80 (range, 0.50 0.97) a 0.82 (range, 0.68 0.95), respectively. A significant deviation from Hardy-Weinberg equilibrium was observed at three loci (KTh6B, KTh10, and KTh16). This high variability indicates that these microsatellites may be useful for high-resolution studies of population genetics.

Intraspecific Relationship Analysis of Eleutherococcus senticosus Max. by RAPD Markers (RAPD 분석에 의한 가시오갈피의 유연관계 분석)

  • 임정대;성은수;최강준;김승경;김명조;유창연
    • Korean Journal of Plant Resources
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    • v.13 no.2
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    • pp.104-110
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    • 2000
  • To analyse the genetic relationship and intraspecific variations among the Eleutherococcus senticosus population, the polymerase chain reaction(PCR) was performed total genomic DNAs of 10 E. senticosus collections by random 10 primers. The genetic diversity and genetic distance among 10 collections of Eleutherococcus spp. were used to describe the dendrogram showing phylogenic relationship. Ten collections were classfied into two group(group I, II) at the similarity coefficient value of 0.50. Group I included E. senticosus of Bukhado(Japanese), youngwal(Korea), E. seoulense, and E. chiisanesis while group II included several internal and Russia collection. The range of polymorphism was from 66.7 to 90.9% in 87 amplified DNA fragments. The similarity value of all collections ranged from 0.41 to 0.92. The average of genetic distance was 0.61.

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Genetic analysis of endangered species Crested Ibis (Nipponia nippon) microsatellite markers (Microsatellite Markers를 이용한 따오기의 유전적 특성 분석)

  • Kim, Da Hye;Kim, Yi Seul;Seo, Joo Hee;Kim, Sung Jin;Kong, Hong Sik
    • Korean Journal of Ornithology
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    • v.25 no.2
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    • pp.77-81
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    • 2018
  • The Korean Crested ibis Nipponia Nippon is an endangered species. A pair of Crested ibis was introduced from China in October 2008, and a successful program of artificial incubation of the species, and over 200 animals have been successfully bred through the restoration project up to 2017 at Upo ibis restoration center. We assessed genetic diversity and sex determination in the Korean Crested ibis. In total, 228 Crested ibis (115 females and 113 males) were identified. And genetic diversity measures, observed heterozygosity, expected heterozygosity, and polymorphic information content values were lower in 2017 than those in 2016. The inbreeding coefficient showed that the degree of ancestry increased in 2017. The decrease in polymorphism and increase in the degree of ancestry is thought to be due to inbreeding in such a small group. In this study provided important insight into protocols for genetic management of the breeding population of Korean Crested ibis in Korea and will help in extending the restoration program.

Development of a Molecular Marker Linked to the A4 Locus and the Structure of HD Genes in Pleurotus eryngii

  • Lee, Song Hee;Ali, Asjad;Ha, Byeongsuk;Kim, Min-Keun;Kong, Won-Sik;Ryu, Jae-San
    • Mycobiology
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    • v.47 no.2
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    • pp.200-206
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    • 2019
  • Allelic differences in A and B mating-type loci are a prerequisite for the progression of mating in the genus Pleurotus eryngii; thus, the crossing is hampered by this biological barrier in inbreeding. Molecular markers linked to mating types of P. eryngii KNR2312 were investigated with randomly amplified polymorphic DNA to enhance crossing efficiency. An A4-linked sequence was identified and used to find the adjacent genomic region with the entire motif of the A locus from a contig sequenced by PacBio. The sequence-characterized amplified region marker $7-2_{299}$ distinguished A4 mating-type monokaryons from KNR2312 and other strains. A BLAST search of flanked sequences revealed that the A4 locus had a general feature consisting of the putative HD1 and HD2 genes. Both putative HD transcription factors contain a homeodomain sequence and a nuclear localization sequence; however, valid dimerization motifs were found only in the HD1 protein. The ACAAT motif, which was reported to have relevance to sex determination, was found in the intergenic region. The SCAR marker could be applicable in the classification of mating types in the P. eryngii breeding program, and the A4 locus could be the basis for a multi-allele detection marker.

Genetic diversity of grapevine (Vitis vinifera L.) as revealed by ISSR markers

  • Basheer-Salimia, Rezq;Mujahed, Arwa
    • Journal of Plant Biotechnology
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    • v.46 no.1
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    • pp.1-8
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    • 2019
  • The main goal of this study was to determine the genetic diversity among 36 grape cultivars grown in Palestine by using ISSR-polymerase chain reaction (PCR) fingerprints. Among the tested primers, 17 produced reasonable amplification products with high intensity and pattern stability. A total of 57 DNA fragments (loci) separated by electrophoresis on agarose gels were detected and they ranged in size, from 150 to 900 bp. Out of these fragments, 55 (88%) were polymorphic and 2 (3.5%) monomorphic. Our results also revealed an average of 3.1 loci per primer. A minimum of 1 and maximum of 10 DNA fragments were obtained (S-17, #820 and #841) and (S-31) primers, respectively. Therefore, the later primer (S-31) is considered to be the most powerful primer among the tested ones. The genetic distance matrix showed an average distance range of between 0.05 and 0.76. The maximum genetic distance value of 0.76 (24% similarity) was exhibited between the (Shami and Marawi.Hamadani.Adi) as well as (Bairuti and Marawi.Hamadani.Adi) genotypes. On the other hand, the lowest genetic distance of 0.05 (95% similarity) was exhibited between (Jandali.Tawel.Mofarad and Jandali. Kurawi.Mlzlz) along with (Shami.Aswad and Shami.mtartash. mlwn) genotypes. Furthermore, the UPGMA dendrogram generally clusters the grape cultivars into eight major clusters in addition to an isolated genotype. Based on these figures, the cultivars tested in this study could be characterized by large divergence at the DNA level. This is taking the assumption that our region has a very rich and varied clonal grape genetic structure.

Composite genotypes of progestogen-associated endometrial protein gene and their association with composition and quality of dairy cattle milk

  • Kolenda, Magdalena;Sitkowska, Beata;Kamola, Dariusz;Lambert, Barry D.
    • Animal Bioscience
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    • v.34 no.8
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    • pp.1283-1289
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    • 2021
  • Objective: The progestogen-associated endometrial protein (PAEP) gene encodes the main whey protein in milk, β-lactoglobulin. The aim of the study was to investigate polymorphism in the PAEP gene and its association with milk yield, composition, and quality. Methods: Test-day records for 782 dairy cows were analysed. A total of 10 single nucleotide polymorphisms (SNP) within the PAEP gene were investigated. The following parameters were recorded: milk yield (MY, kg/d), percent milk fat (%), protein (PP, %), dry matter (DMP, %) and lactose (LP, %), urea content (UC, mg/L) as well as natural logarithm for somatic cell count (LnSCC, ln). Effect on genomic estimated breeding values accuracy was evaluated with pedigree and single step model. Results: Results show that only three SNPs were polymorphic, creating 5 composite genotypes: P1 to P5. Differences in MY between composite genotypes were noted in the two tested herds. Cows with P5 composite genotypes were characterised by the highest PP and LnSCC and the lowest LP and UC (p<0.05). P4 was linked to an increased DMP and UC, while P3 to an increase in LP and decrease in PP and LnSCC. Both factors are important markers in herd management and have high influences on the herds economics. For 5 out of 7 traits the accuracy of prediction was improved by including the haplotype as a fixed effect. Conclusion: Presented results may suggest a new way to optimise breeding programmes and demonstrate the impact of using genomic data during that process.

Microsatellite Markers for Non-Invasive Examination of Individual Identity, Genetic Variation, and Population Differentiation in Two Populations of Korean Long-Tailed Goral (Naemorhedus caudatus)

  • Kim, Baek-Jun
    • Proceedings of the National Institute of Ecology of the Republic of Korea
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    • v.3 no.4
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    • pp.191-198
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    • 2022
  • Natural habitats of the Korean long-tailed goral (Naemorhedus caudatus) have been fragmented by anthropogenic activities in South Korea in the last decades. Here, the individual identity, genetic variation, and population differentiation of the endangered species were examined via the multiple-tube approach using a non-invasive genotyping method. The average number of alleles was 3.16 alleles/locus for the total population. The Yanggu population (1.66) showed relatively lower average number of alleles than the Inje population (3.67). Of the total 19 alleles, only seven (36.8%) alleles were shared by the two populations. Using five polymorphic out of six loci, four and six different goral individuals from the captive Yanggu (n=24) and the wild Inje (n=28) population were identified, respectively. The allele distribution was not identical between the two populations (Fisher's exact test: P<0.01). A considerably low migration rate was detected between the two populations (no. of migrants after correction for size=0.294). Additionally, the F statistics results indicated significant population differentiation between them, however, quite low (FST=0.327, P<0.01). The posterior probabilities indicated that the two populations originated from a single panmictic population (P=0.959) and the assignment test results designated all individuals to both populations with nearly equal likelihood. These could be resulted from moderate population differentiation between the populations. No significant evidence supported recent population bottleneck in the total Korean goral population. This study could provide us with useful population genetic information for conservation and management of the endangered species.

Molecular Identification and Effects of Temperature on Survival and Growth of Hybrids between Haliotis gigantea Gmelin (♀) and Haliotis discus hannai Reeve (♂)

  • An, Hye Suck;Han, Jong Won;Hwang, Hyun-Ju;Jeon, Hancheol;Jung, Seung-Hyun;Jo, Seonmi;Choi, Tae-Young;Hyun, Young Se;Song, Ha Yeun;Whang, Ilson
    • Journal of Marine Life Science
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    • v.2 no.2
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    • pp.83-89
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    • 2017
  • In abalones, interspecific hybridization has been suggested as a possible means to increase production and desired traits for the industry. In Korea, Haliotis gigantea is considered a species with a larger size and higher temperature tolerance than H. discus hannai. However, H. discus hannai is considered the most valuable and popular fishery resource due to its better acceptance and higher market prices. Thus, viable interspecific hybrids have been produced by artificial inseminating H. gigantea eggs with H. discus hannai sperm. However, the reciprocal hybrid cross was not successful. In this study, the hybridity and the growth and thermal tolerance performance of the interspecific hybrids were examined. A combination of various assays revealed maximum growth occurrence at 21℃ and the higher growth rate in the hybrids than that of H. discus hannai parent. In addition, the growth and survival at high-temperature (28℃) of the hybrids was equivalent to that of the highly tolerant H. gigantea parent, suggesting new possibilities to overcome the mass mortality in H. discus hannai during high temperature periods of summer season in Korea. Furthermore, the induced interspecific hybrid status was confirmed by the presence of species-specific bands for each parental species of the random amplified polymorphic DNA (RAPD) profiles using universal rice primer (URP), which could be used as speciesspecific markers to distinguish the hybrids and their parental species.