• Title/Summary/Keyword: Genome Analysis

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Cytogenetic Analysis of Reciprocal Hybrids Reveals a Robertsonian Translocation between Mud Loach ($Misgurnus$ $mizolepis$) and Cyprinid Loach ($M.$ $anguillicaudatus$) (미꾸라지($Misgurnus$ $mizolepis$)와 미꾸리($M.$ $anguillicaudatus$) 및 유도된 종간 잡종의 세포유전학적 연구)

  • Lee, Seung-Ki;Kim, Dong-Soo
    • Korean Journal of Ichthyology
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    • v.24 no.1
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    • pp.1-10
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    • 2012
  • Reciprocal hybrids between the mud loach ($Misgurnus$ $mizolepis$) and cyprinid loach ($M.$ $anguillicaudatus$) were produced by artificial fertilization. The chromosome number of mud loach was 2n=48, consisting of 12M+4SM+32A chromosomes. The cyprinid loach has 2n=50, consisting of 10M+4SM+36A chromosomes. The chromosome numbers of the diploid reciprocal hybrids were 2n=49, consisting of 11M+4SM+34A chromosomes. All the karyotypes documented in this study had the same arm number of 64. There was no evidence of chromosomal polymorphisms or sex-related heteromorphism. The cytogenetic traits of the hybrid genotypes were intermediate between those of the parent species. In all genotypes, the chromosomal NORs localized to the terminal short arms of the same metacentric chromosome pair. These results suggest that Robertsonian translocation occurred between metacentric chromosome 1 of mud loach and acrocentric chromosome of cyprinid loach.

Purification and Characterization of HCV RNA-dependent RNA Polymerase from Korean Genotype 1b Isolate: Implications for Discovery of HCV Polymerase Inhibitors

  • Kim, Jeong-Min;Lee, Mi-Kyoung;Kim, Yong-Zu
    • Bulletin of the Korean Chemical Society
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    • v.26 no.2
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    • pp.285-291
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    • 2005
  • The nonstructural protein 5B (NS5B) of hepatitis C virus (HCV) is the viral RNA-dependent RNA polymerase (RdRp), which is the essential catalytic enzyme for the viral replication and is an appealing target for the development of new therapeutic agents against HCV infection. A small amount of serum from a single patient with hepatitis C was used to get the genome of a Korean HCV isolate. Sequence analysis of NS5B 1701 nucleotides showed the genotype of a Korean isolate to be subtype 1b. The soluble recombinant HCV NS5B polymerase lacking the C-terminal 24 amino acids was expressed and purified to homogeneity. With the highly purified NS5B protein, we established in vitro systems for RdRp activity to identify potential polymerase inhibitors. The rhodanine family compounds were found to be potent and specific inhibitors of NS5B from high throughput screening (HTS) assay utilizing the scintillation proximity assay (SPA) system. The binding mode of an inhibitor was analyzed by measuring various kinetic parameters. Lineweaver-Burk plots of the inhibitor suggested it binds not to the active site of NS5B polymerase, but to an allosteric site of the enzyme. The activity of NS5B in in vitro polymerase reactions with homopolymeric RNA requires interaction with multiple substrates that include a template/primer and ribonucleotide triphosphate. Steady-state kinetic parameter, such as Km, was determined for the ribonucleotide triphosphate. One of compounds found interacts directly with the viral polymerase and inhibits RNA synthesis in a manner noncompetitively with respect to UTP. Furthermore, we also investigated the ability of the compound to inhibit NS5B-directed viral RNA replication using the Huh7 cell-based HCV replicon system. The investigation is potentially very useful for the utility of such compounds as anti-hepatitic agents.

Identification of the quantitative trait loci (QTL) for seed protein and oil content in soybean.

  • Jeong, Namhee;Park, Soo-Kwon;Ok, Hyun-Choong;Kim, Dool-Yi;Kim, Jae-Hyun;Choi, Man-Soo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.148-148
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    • 2017
  • Soybean is an important economical resource of protein and oil for human and animals. The genetic basis of seed protein and oil content has been separately characterized in soybean. However, the genetic relationship between seed protein and oil content remains to be elucidated. In this study, we used a combined analysis of phenotypic correlation and linkage mapping to dissect the relationship between seed protein and oil content. A $F_{10:11}$ RIL population containing 222 lines, derived from the cross between two Korean soybean cultivars Seadanbaek as female and Neulchan as male parent, were used in this experiment. Soybean seed analyzed were harvested in three different experimental environments. A genetic linkage map was constructed with 180K SoyaSNP Chip and QTLs of both traits were analyzed using the software QTL IciMapping. QTL analyses for seed protein and oil content were conducted by composite interval mapping across a genome wide genetic map. This study detected four major QTL for oil content located in chromosome 10, 13, 15 and 16 that explained 13.2-19.8% of the phenotypic variation. In addition, 3 major QTL for protein content were detected in chromosome 10, 11 and 16 that explained 40.8~53.2% of the phenotypic variation. A major QTLs was found to be associated with both seed protein and oil content. A major QTL were mapped to soybean chromosomes 16, which were designated qHPO16. These loci have not been previously reported. Our results reveal a signi cant genetic relationship between seed protein and oil fi content traits. The markers linked closely to these major QTLs may be used for selection of soybean varieties with improved seed protein and oil content.

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Current status of wheat allergy research and prospect

  • Son, Jae-Han;Yoon, Young-mi;Cheong, Young-Keun;Park, Jong-Chul;Kim, Kyong-Ho;Kim, Bo-Kyeong;Kang, Chon-Sik
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.259-259
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    • 2017
  • Wheat is one of important food crop in the world. However, wheat has some negative things that are allergen. Wheat includes four different allergen kwon as alpha-, beta- and gamma gliadin and amylase inhibitor. Recently, people are interesting to gluten free food or low gliadin food. To study about wheat allergies and research trend is important for future research like as wheat breeding and allergy reduction technology. In this study, we analyzed that the reported on the research field of wheat allergy have been reported until now, and was to provide direction for future research. The analysis of 235 episodes of major papers published from 2007 to 2016. The last 5 years from 2012, wheat allergy appears increasing number of related papers. The ratio of national papers 13% and 60% were published by the United States and other European countries, respectively. The fields of wheat allergen-related preclinical technology and wheat allergen related to genome research for discovery technology were represented high rate by 58% and 26%, respectively. In the case of Korea, significantly account genetic and breeding areas. Recently, however, the research of glutenin protein which is closely linked to wheat allergy is in progress. So, we expected to increase wheat allergy of the research is in the future in Korea.

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Positional mapping for foxglove aphid resistance with 180k SNP array in soybean [Glycine max (L.) Merr.]

  • Park, Sumin;Kim, Kyung Hye;Go, Hong Min;Lee, Ju Seok;Jung, Jin Kyo;Bilyeu, Kristin D.;Lee, Jeong-Dong;Kan, Sungtaeg
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.145-145
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    • 2017
  • Foxglove aphid, Aulacorthum solani (Kaltenbach), is a Hemipteran insect that infected a wide variety of plants worldwide and caused serious yield losses in crops. The objective of this study was to identify the putative genes to foxglove aphid resistance in wild soybean, PI 366121 (Glycine soja Sieb. and Zucc.). One hundred and forty-one F4:8 recombinant inbred lines developed from a cross between susceptible variety, Williams 82 and foxglove aphid resistance wild soybean, PI 366121 were used. The two type of resistance response, antibiosis and antixenosis resistance were evaluated through choice and no-choice test, graded by the degree of total plant damage and primary infestation leaf damage; a genome-wide molecular linkage map was constructed with 29,898 single-nucleotide polymorphism markers utilizing a Axiom(R) 180K soyaSNP array. Using inclusive composite interval mapping analysis for foxglove aphid resistance, one major candidate QTL on chromosome 7 was identified. The major QTL on chromosome 7 showed both antixenosis and antibiosis resistance responses. The newly identified major QTL was consistent with previously reported QTL, Raso2, which showed around 5 times narrow down interval range with 8 candidate genes. Furthermore, total 1,115 soybean varieties including Glycine soja and Glycine max were exposed to germplasm screening, and 31 varieties, which showed significant antibiosis type foxglove aphid resistance were identified. This result could be useful in breeding for new foxglove aphid resistant soybean cultivars and developing novel insecticides.

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Sorghum TCP transcription factor MULTISEED1 affects grain yield regulating at pedicellate spikelet fertility

  • Lee, Young Koung;Jiao, Yinping;Gladman, Nicholas;Chopra, Ratan;Burow, Gloria;Burke, John;Xin, Zhanguo;Ware, Doreen
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.25-25
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    • 2017
  • Inflorescence architecture mainly contributes to final grain yield in crops. Sorghum inflorescence is basically composed of one fertile sessile spikelet (SS) and two infertile pedicellate spikelets (PS). To identify regulatory factors involved in the inflorescence architecture, we screened an EMS mutagenesis population from the pedigreed sorghum mutant library. We found inflorescent architecture mutants, named as multi-seed mutants, msd, with gained fertile ability in PS and also an increased number of floral branches. In natural sorghum populations, it is not common that are fertile. A detailed dissection of developmental stages of wild type and msd1 mutant described that the PS in wild type do not have floral organs, including ovary, stigma, filament and anther, while the msd1 mutants generate intact floral organ in the sessile spikelet. We found MSD1 encoded a TCP transcription factor using bulk segregant analysis (BSA) of F2 population, and was a strongly enriched expression during inflorescence developmental stages. We proposed that MSD1 functions to suppress floral organ maintenance at PS during inflorescence development in Sorghum. To explore the regulatory network associated with PS fertility, whole genome expression profiling was performed at 4 different developmental stages in 6 various tissue types between wild type and msd1. Taken together, we demonstrated that MSD1 was involved in the plant hormone and maybe influenced program cell death in PS via the activation of plant hormonal pathway.

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Molecular cloning, expression and characterization of a squalene synthase gene from grain amaranth (Amaranthus cruentus L.)

  • Park, Young-Jun;Nemoto, Kazuhiro;Matsushima, Kenichi;Um, Han-Yong;Choi, Jung-Hoon;Oh, Chan-sung;Nishikawa, Tomotaro
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.89-89
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    • 2017
  • A gene encoding squalene synthase from grain amaranth was cloned and characterized. The full-length cDNA was 1805-bp long and contained a 1248-bp open reading frame encoding a protein of 416 amino acids with a molecular mass of 47.6 kDa. Southern blot analysis revealed that the A. cruentus genome contained a single copy of the gene. Comparison of the cDNA and genomic sequences indicated that the amaranth SQS gene had 12 introns and 13 exons. All of the exons contributed to the coding sequence. The predicted amino acid sequence of the SQS cDNA shared high homology with those of SQSs from several other plants. It contained conserved six domains that are believed to represent crucial regions of the active site. We conducted qRT-PCR analyses to examine the expression pattern of the SQS gene in seeds at different developmental stages and in several tissues. The amaranth SQS gene was low levels of SQS transcripts at the initial stage of seed development, but the levels increased rapidly at the mid-late developmental stages before declining at the late developmental stage. These findings showed that the amaranth SQS is a late-expressed gene that is rapidly expressed at the mid-late stage of seed development. In addition, we observed that the SQS mRNA levels in stems and roots increased rapidly during the four- to six-leaf stage of development. Therefore, our results showed that the expression levels of SQS in stem and root tissues are significantly higher than those in leaf tissues. In present study provides useful information about the molecular characterization of the SQS clone isolated from grain amaranth. Finally, a basic understanding of these characteristics will contribute to further studies on the amaranth SQS.

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Application of the Molecular Marker in Linkage Disequilibrium with Ms, a Restorer-of-fertility Locus, for Improvement of Onion Breeding Efficiency

  • Kim, Sujeong;Kim, Sunggil
    • Horticultural Science & Technology
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    • v.33 no.4
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    • pp.550-558
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    • 2015
  • To analyze the linkage relationships among molecular markers recently reported to be linked to onion (Allium cepa L.) Ms, a restorer-of-fertility locus, in onion (Allium cepa L.), three single nucleotide polymorphism markers were converted into cleaved amplified polymorphic sequence (CAPS) markers based on onion transcriptome sequences and the rice genome database. Analysis of the recombinants selected from 4,273 segregating plants using CAPS and other linked markers demonstrated the jnurf13 and jnurf610 markers to perfectly co-segregate with the Ms locus. In contrast to jnurf13, the jnurf610 marker was not in perfect linkage disequilibrium with the Ms locus in diverse breeding lines. Thus, the jnurf13 marker and the marker for identification of cytoplasm types were utilized to enhance the efficiency of onion breeding through four applications. First, 89 maintainer lines containing the normal cytoplasm and homozygous recessive Ms genotypes were successfully identified from 100 breeding lines. Second, these two molecular markers were used to analyze the main sources of male-fertile contaminants frequently found in the male-sterile parental lines during F1 hybrid seed production. The majority of the contaminants contained heterozygous Ms genotypes, indicating that pollen grains harboring the dominant Ms genotype may have been introduced during propagation of the maintainer lines. Therefore, the genetic purity of the two maintainer lines was analyzed in the third application, and the results showed that both maintainer lines contained 13-21% off-types. Finally, the two markers were used to increase the seed yield potentials of two open-pollinated varieties containing sterile cytoplasms by removing the plants harboring homozygous recessive and heterozygous Ms genotypes.

Acute Toxicity Evaluation to Daphnia magna of Disease Resistant(OsCK1) Rice (병저항성 GM(OsCK1)벼의 물벼룩(Daphnia magna)에 대한 급성독성 평가)

  • Oh, Sung-Dug;Lee, Kijong;Park, Soo-Yun;Ryu, Tae-Hun;Suh, Sang Jae
    • Korean Journal of Environmental Agriculture
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    • v.33 no.2
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    • pp.121-128
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    • 2014
  • BACKGROUND: The disease resistant (OsCK1) rice was generated by inserting choline kinase (CK1) and phosphinothricin acetyltransferase (PAT) genes isolated from Oryza sativa and Streptomyces hygroscopicus into the genome of the rice, Nakdongbyeo. With the potential problems of safeties, the evaluations on non-target organisms are essentially required for the environmental risk assessment of genetically modified (GM) crops. In the present study, we conducted the evaluation of acute toxicity on Daphnia magna that commonly used as a model organism in ecotoxicological studies for non-target organism evaluation. METHODS AND RESULTS: Effect of acute toxicity to Daphnia magna by each concentration were investigated in the disease resistant (OsCK1) rice and non-genetically modified (non-GM) rice, Nakdongbyeo, as concentration (0, 1,000, 1,800, 3,240, 5,830, 10,500 and 20,000 mg/L). The OsCK1 rice used for the test was confirmed to express the OsCK1/PAT gene by the PCR(Polymerase chain reaction) and western blot analysis. Feeding test showed that no significant differences in cumulative immobility and abnormal response of Daphnia magna fed on OsCK1 rice or non-GM rice. The 48hr-$EC_{50}$ values showed no difference between OsCK1 rice (3,147.18 mg/L) and non-GM rice (3,596.27 mg/L). CONCLUSION: This result suggested that there was no significant difference in toxicity to Daphnia magna between OsCK1 rice and non-GM counterpart.

Development of the Large-Scale Oligonucleotide Chip for the Diagnosis of Plant Viruses and its Practical Use

  • Nam, Moon;Kim, Jeong-Seon;Lim, Seungmo;Park, Chung Youl;Kim, Jeong-Gyu;Choi, Hong-Soo;Lim, Hyoun-Sub;Moon, Jae Sun;Lee, Su-Heon
    • The Plant Pathology Journal
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    • v.30 no.1
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    • pp.51-57
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    • 2014
  • A large-scale oligonucleotide (LSON) chip was developed for the detection of the plant viruses with known genetic information. The LSON chip contains two sets of 3,978 probes for 538 species of targets including plant viruses, satellite RNAs and viroids. A hundred forty thousand probes, consisting of isolate-, species- and genus-specific probes respectively, are designed from 20,000 of independent nucleotide sequence of plant viruses. Based on the economic importance, the amount of genome information, and the number of strains and/or isolates, one to fifty-one probes for each target virus are selected and spotted on the chip. The standard and field samples for the analysis of the LSON chip have been prepared and tested by RT-PCR. The probe's specific and/or nonspecific reaction patterns by LSON chip allow us to diagnose the unidentified viruses. Thus, the LSON chip in this study could be highly useful for the detection of unexpected plant viruses, the monitoring of emerging viruses and the fluctuation of the population of major viruses in each plant.