• 식물분류학회지
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• 제53권1호
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• pp.32-37
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• 2023

Spiraea prunifolia f. simpliciflora Nakai is a perennial shrub widely used for horticultural and medicinal purposes. We simultaneously obtained the complete plastid genome (plastome) and nuclear ribosomal gene transcription units, 45S nuclear ribosomal DNA (nrDNA) and 5S nrDNA of S. prunifolia f. simpliciflora, using Illumina short-read data. The plastome is 155,984 bp in length with a canonical quadripartite structure consisting of 84,417 bp of a large single-copy region, 18,887 bp of a short single-copy region, and 26,340 bp of two inverted repeat regions. Overall, a total of 113 genes (79 protein-coding genes, 30 tRNAs, and four rRNAs) were annotated in the plastome. The 45S nrDNA transcription unit is 5,848 bp in length: 1,809 bp, 161 bp, and 3,397 bp for 18S, 5.8S, and 26S, respectively, and 261 bp and 220 bp for internal transcribed spacer (ITS) 1 and ITS 2 regions, respectively. The 5S nrDNA unit is 512 bp, including 121 bp of 5S rRNA and 391 bp of intergenic spacer regions. Phylogenetic analyses showed that the genus Spiraea was monophyletic and sister to the clade of Sibiraea angustata, Petrophytum caespitosum and Kelseya uniflora. Within the genus Spiraea, the sections Calospira and Spiraea were monophyletic, but the sect. Glomerati was nested within the sect. Chamaedryon. In the sect. Glomerati, S. prunifolia f. simpliciflora formed a subclade with S. media, and the subclade was sister to S. thunbergii and S. mongolica. The close relationship between S. prunifolia f. simpliciflora and S. media was also supported by the nrDNA phylogeny, indicating that the plastome and nrDNA sequences assembled in this study belong to the genus Spiraea. The newly reported complete plastome and nrDNA transcription unit sequences of S. prunifolia f. simpliciflora provide useful information for further phylogenetic and evolutionary studies of the genus Spiraea, as well as the family Rosaceae.

• 한국환경농학회지
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• 제41권4호
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• pp.335-343
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• 2022

BACKGROUND: Antibiotics used in animal husbandry for disease prevention and treatment have resulted in the rapid progression of antibiotic resistant bacteria which can be introduced into the environment through livestock feces/manure, disseminating antibiotic resistant genes (ARGs). In this study, fecal samples were collected from the livestock farms located in Jeju Island to investigate the relationship between microbial communities and ARGs. METHODS AND RESULTS: Illumina MiSeq sequencing was applied to characterize microbial communities within each fecal sample. Using quantitative PCR (qPCR), ten ARGs encoding tetracycline resistance (tetB, tetM), sulfonamide resistance (sul1, sul2), fluoroquinolone resistance (qnrD, qnrS), fluoroquinolone and aminoglycoside resistance (aac(6')-Ib), beta-lactam resistance (bla_TEM, bla_CTX-M), macrolide resistance (ermC), a class 1 integronsintegrase gene (intI1), and a class 2 integrons-integrase gene (intI2) were quantified. The results showed that Firmicutes and Bacteroidetes were dominant in human, cow, horse, and pig groups, while Firmicutes and Actinobacteria were dominant in chicken group. Among ARGs, tetM was detected with the highest number of copies, followed by sul1 and sul2. Most of the genera belonging to Firmicutes showed positive correlations with ARGs and integron genes. There were 97, 34, 31, 25, and 22 genera in chicken, cow, pig, human, and horse respectively which showed positive correlations with ARGs and integron genes. In network analysis, we identified diversity of microbial communities which correlated with ARGs and integron genes. CONCLUSION(S): In this study, antibiotic resistance patterns in human and livestock fecal samples were identified. The abundance of ARGs and integron genes detected in the samples were associated with the amount of antibiotics commonly used for human and livestocks. We found diverse microbial communities associated with antibiotics resistance genes in different hosts, suggesting that antibiotics resistance can disseminate across environments through various routes. Identifying the routes of ARG dissemination in the environment would be the first step to overcome the challenge of antibiotic resistance in the future.

• Animal Bioscience
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• 제35권9호
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• pp.1303-1313
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• 2022

Objective: The current study aimed to perform whole-genome resequencing of Chinese indigenous Mongolian sheep breeds including Ujimqin, Sunit, and Wu Ranke sheep breeds (UJMQ, SNT, WRK) and deeply analyze genetic variation, population structure, domestication, and selection for domestication traits among these Mongolian sheep breeds. Methods: Blood samples were collected from a total of 60 individuals comprising 20 WRK, 20 UJMQ, and 20 SNT. For genome sequencing, about 1.5 ㎍ of genomic DNA was used for library construction with an insert size of about 350 bp. Pair-end sequencing were performed on Illumina NovaSeq platform, with the read length of 150 bp at each end. We then investigated the domestication and signatures of selection in these sheep breeds. Results: According to the population and demographic analyses, WRK and SNT populations were very similar, which were different from UJMQ populations. Genome wide association study identified 468 and 779 significant loci from SNT vs UJMQ, and UJMQ vs WRK, respectively. However, only 3 loci were identified from SNT vs WRK. Genomic comparison and selective sweep analysis among these sheep breeds suggested that genes associated with regulation of secretion, metabolic pathways including estrogen metabolism and amino acid metabolism, and neuron development have undergone strong selection during domestication. Conclusion: Our findings will facilitate the understanding of Chinese indigenous Mongolian sheep breeds domestication and selection for complex traits and provide a valuable genomic resource for future studies of sheep and other domestic animal breeding.

• Animal Bioscience
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• 제35권9호
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• pp.1327-1339
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• 2022

Objective: Fat deposition in poultry is an important factor in production performance and meat quality research. miRNAs also play important roles in regulating adipocyte differentiation process. This study was to investigate the expression patterns of miRNAs in duck adipocytes after differentiation and explore the role of miR-214 in regulating carnitine palmitoyltransferases 2 (CPT2) gene expression during duck adipocyte differentiation. Methods: Successful systems for the isolation, culture, and induction of duck primary fat cells was developed in the experiment. Using Illumina next-generation sequencing, the miRNAs libraries of duck adipocytes were established. miRanda was used to predict differentially expressed (DE) miRNAs and their target genes. The expression patterns of miR-214 and CPT2 during the differentiation were verified by quantitative real-time polymerase chain reaction and western blot. Luciferase reporter assays were used to explore the specific regions of CPT2 targeted by miR-214. We used a miR-214 over-expression strategy in vitro to further investigate its effect on differentiation process and CPT2 gene transcription. Results: There were 481 miRNAs identified in duck adipocytes, included 57 DE miRNA candidates. And the 1,046 targets genes of DE miRNAs were mainly involved in p53 signaling, FoxO signaling, and fatty acid metabolism pathways. miR-214 and CPT2 showed contrasting expression patterns before and after differentiation, and they were selected for further research. The expression of miR-214 was decreased during the first 3 days of duck adipocytes differentiation, and then increased, while the expression of CPT2 increased both in the transcriptional and protein level. The luciferase assay suggested that miR-214 targets the 3'untranslated region of CPT2. Overexpression of miR-214 not only promoted the formation of lipid droplets but also decreased the protein abundance of CPT2. Conclusion: Current study reports the expression profile of miRNAs in duck adipocytes differentiated for 4 days. And miR-214 has been proved to have the regulator potential for fat deposition in duck.

• Journal of Animal Science and Technology
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• 제65권4호
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• pp.735-747
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• 2023

The composition of fatty acids determines the flavor and quality of meat. Flavor compounds are generated during the cooking process by the decomposition of volatile fatty acids via lipid oxidation. A number of research on candidate genes related to fatty acid content in livestock species have been published. The majority of these studies focused on pigs and cattle; the association between fatty acid composition and meat quality in chickens has rarely been reported. Therefore, this study investigated candidate genes associated with fatty acid composition in chickens. A genome-wide association study (GWAS) was performed on 767 individuals from an F2 crossbred population of Yeonsan Ogye and White Leghorn chickens. The Illumina chicken 60K significant single-nucleotide polymorphism (SNP) genotype data and 30 fatty acids (%) in the breast meat of animals slaughtered at 10 weeks of age were analyzed. SNPs were shown to be significant in 15 traits: C10:0, C14:0, C18:0, C18:1n-7, C18:1n-9, C18:2n-6, C20:0, C20:2, C20:3n-6, C20:4n-6, C20:5n-3, C24:0, C24:1n-9, monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA). These SNPs were mostly located on chromosome 10 and around the following genes: ACSS3, BTG1, MCEE, PPARGC1A, ACSL4, ELOVL4, CYB5R4, ME1, and TRPM1. Both oleic acid and arachidonic acid contained the candidate genes: MCEE and TRPM1. These two fatty acids are antagonistic to each other and have been identified as traits that contribute to the production of volatile fatty acids. The results of this study improve our understanding of the genetic mechanisms through which fatty acids in chicken affect the meat flavor.

• Animal Bioscience
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• 제37권5호
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• pp.795-806
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• 2024

Objective: Karan Fries (KF), a high-producing composite cattle was developed through crossing indicine Tharparkar cows with taurine bulls (Holstein Friesian, Brown Swiss, and Jersey), to increase the milk yield across India. This composite cattle population must maintain sufficient genetic diversity for long-term development and breed improvement in the coming years. The level of linkage disequilibrium (LD) measures the influence of population genetic forces on the genomic structure and provides insights into the evolutionary history of populations, while the decay of LD is important in understanding the limits of genome-wide association studies for a population. Effective population size (N_e) which is genomically based on LD accumulated over the course of previous generations, is a valuable tool for e valuation of the genetic diversity and level of inbreeding. The present study was undertaken to understand KF population dynamics through the estimation of N_e and LD for the long-term sustainability of these breeds. Methods: The present study included 96 KF samples genotyped using Illumina HDBovine array to estimate the effective population and examine the LD pattern. The genotype data were also obtained for other crossbreds (Santa Gertrudis, Brangus, and Beefmaster) and Holstein Friesian cattle for comparison purposes. Results: The average LD between single nucleotide polymorphisms (SNPs) was r² = 0.13 in the present study. LD decay (r² = 0.2) was observed at 40 kb inter-marker distance, indicating a panel with 62,765 SNPs was sufficient for genomic breeding value estimation in KF cattle. The pedigree-based N_e of KF was determined to be 78, while the N_e estimates obtained using LD-based methods were 52 (SNeP) and 219 (genetic optimization for N_e estimation), respectively. Conclusion: KF cattle have an N_e exceeding the FAO's minimum recommended level of 50, which was desirable. The study also revealed significant population dynamics of KF cattle and increased our understanding of devising suitable breeding strategies for long-term sustainable development.

• BMB Reports
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• 제57권3호
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• pp.161-166
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• 2024

Aberrant DNA methylation plays a critical role in the development and progression of colorectal cancer (CRC), which has high incidence and mortality rates in Korea. Various CRC-associated methylation markers for cancer diagnosis and prognosis have been developed; however, they have not been validated for Korean patients owing to the lack of comprehensive clinical and methylome data. Here, we obtained reliable methylation profiles for 228 tumor, 103 adjacent normal, and two unmatched normal colon tissues from Korean patients with CRC using an Illumina Infinium EPIC array; the data were corrected for biological and experiment biases. A comparative methylome analysis confirmed the previous findings that hypermethylated positions in the tumor were highly enriched in CpG island and promoter, 5' untranslated, and first exon regions. However, hypomethylated positions were enriched in the open-sea regions considerably distant from CpG islands. After applying a CpG island methylator phenotype (CIMP) to the methylome data of tumor samples to stratify the CRC patients, we consolidated the previously established clinicopathological findings that the tumors with high CIMP signatures were significantly enriched in the right colon. The results showed a higher prevalence of microsatellite instability status and MLH1 methylation in tumors with high CMP signatures than in those with low or non-CIMP signatures. Therefore, our methylome analysis and dataset provide insights into applying CRC-associated methylation markers for Korean patients regarding cancer diagnosis and prognosis.

• 한국초지조사료학회지
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• 제38권2호
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• pp.106-111
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• 2018

본 연구는 고창증 발병으로 폐사된 저지종 젖소의 반추위액과 캐놀라가 장착한 저지종 젖소의 반추위액을 샘플링하여 반추위 내 미생물 균총의 급작스런 변화에 따른 반추위 발효와 고창증 발병간에 연관성을 알아보고자 진행되었다. 채취한 반추위액 샘플은 PowerMax Soil DNA Isolation Kit (MO BIO Inc., CA, USA)를 이용하여 DNA를 추출한 후 PicoGreen (Turner BioSystems, Inc., CA, USA)과 Nanodrop spectrophotometer (ND-1000, NanoDrop Technologies Inc., DE, USA)를 이용하여 260/280 nm와 260/230 nm 흡광도 값을 측정하였다. DNA를 정량분석하고 $MiSeq^{TM}$ platform (Illumina, CA, USA)을 이용한 장내 미생물 균총의 다양성 분석을 마크로젠(Macrogen Inc., Seoul)에 의뢰하여 실시하였다. 반추위액 내 미생물 균총 분석 결과, 전분 분해균의 경우 고창증 발병으로 인한 폐사우의 반추위 내에서 R. bromii가 우점 하였으며, R. bromii, B. pseudolongum, B. merycicum 및 B. fibrisolvens 등과 같은 전분 분해균들이 정상우 대비 36배 높았다. 반면 반추위 발효와 밀접한 관련이 있는 F. succinogenes, R. albus 및 R. flavefaciens 등과 같은 대표적인 섬유소 분해균 비율은 정상우에서 고창증 발병으로 인한 폐사우 대비 12배 높았다. 이와 같은 결과를 볼 때, 반추위 내 균총 가운데 R. bromii, B. pseudolongum, B. merycicum 및 B. fibrisolvens 등과 같은 전분 분해균들의 급격한 증가가 반추위 내 pH 저하 및 가스 생성 증가를 초래했고 이는 저지종 젖소의 고창증 발병으로 인한 폐사와 밀접한 연관이 있었을 것으로 판단된다.

• Journal of Animal Science and Technology
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• 제55권1호
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• pp.13-18
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• 2013

본 연구는 농협 한우개량사업소 후대검정우 552두의 도체중, 배최장근단면적, 등지방두께 및 근내지방도를 측정한 후 고밀도 SNP 패널(777K)을 사용하여 유전체 혈연 행렬(Genetic Relationship Matrix, GRM)을 추정하고 GBLUP (Genomic Best Linear Unbiased Prediction) 방법으로 GEBV (Genomic Estimated Breeding Value)를 구하여 교차 검증(Cross-validation) 방법으로 그 정확도를 추정함으로써 유전체 선발 기법을 한우 유전평가 체계에 적용하기 위한 기초자료로 이용하고자 수행하였다. 교차 검증 방법으로 각 형질별로 추정된 유전체 육종가의 정확도는 0.915~0.957로 상당히 높게 추정되었다. 대립유전자의 빈도로 계산된 유전체 혈연 행렬을 이용하여 GBLUP 방법으로 추정된 육종가 정확도의 최대 차이는 후대검정우 534두에 대하여 도체중, 배최장근단면적, 등지방 두께 및 근내지방도 순으로 각각 9.56%, 5.78%, 5.78% 및 4.18% 정도의 수준으로 상승했고, 혈통 기록상의 모든 개체 3,674두에 대해서는 형질 별로 최대 13.54%, 6.50%, 6.50% 및 4.31% 정도의 수준으로 증가한 결과가 추정되었다. 이는 한우 보증씨수소의 선발 시스템에서 아직 표현형 자료를 생산할 수 없는 당대검정 후보축 대한 집단을 조성할 때 유전체 정보를 이용한 사전 선발을 활용하면 기존의 상대적으로 낮았던 육종가의 정확도의 상승 효과와 세대 간격의 단축으로 인하여 유전적 개량량을 증대시킬 수 있을 것으로 기대된다. 본 연구에서 genomic breeding value 추정을 위하여 조성된 집단의 경우는 후대 검정우 집단으로서 개체들 간의 혈연관계가 높으며, 이미 전통적인 BLUP 방법으로도 상당히 높은 정확도를 가진 집단을 이용하였다. 그러나, 현재 한우 집단에 대한 유전체 자료 구축 시 이용할 수 있는 정확한 자료는 후대검정우 집단 외에는 참조 집단을 조성할 수 있는 대안이 없으므로, 지속적인 유전체 검정을 위해서는 다양한 유전적 조성이 구축된 참조 집단을 구축해야 할 것으로 사료된다. 또한 유전체 검정을 통한 정확도 상승효과를 기대하기 위해서 지속적으로 참조 집단의 크기를 늘릴 필요성이 있다.

• 한국산학기술학회논문지
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• 제19권1호
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• pp.537-545
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• 2018

본 연구에서는 메밀의 발아초기에 발현되는 전사체의 다양한 정보 수집을 위해 양절메밀과 대관 3-3호의 RNA를 추출하여 전사체 분석을 수행하였다. 제주산 양절메밀과 대관3-3호의 종자 및 발아 후 12, 24, 36시간별로 total RNA를 추출하고, llumina Hiseq 2000 플랫폼을 사용하여 시퀀싱 하였다. SolexaQA package의 DynamicTrim과 LengthsORT 프로그램으로 이용하여 raw 데이터 분석을 실시한 후, 어셈블리(assembly)와 annotation을 수행하였다. RNA-seq raw 데이터로부터 약 84.2%, 81.5%에 해당하는 16.5Gb, 16.2Gb의 transcriptome 데이터를 확보하였다. 47Mb에 해당하는 43,494개의 대표적인 전사체(representative transcripts)를 확보하였고, 그 중에서 annotation DB와 서열 유사도를 갖는 서열은 23,165개로 확인되었다. 메밀의 representative transcripts 유전자의 유전자 온톨로지(gene ontology) 분석결과, biological process는 metabolic process (49.49%)에서, cellular components는 cell (46.12%)에서, molecular function은 catalyltic activity (80.43%)에서 유전자가 많이 분포되어 있는 것을 확인하였다. 종자의 발아에 관련된 gibberellin receptor GID1C의 경우에는 양절메밀, 대관 3-3호의 발현양이 모두 시간이 지남에 따라 증가되는 것을 확인할 수 있었으며, gibberellin 20-oxidase1의 경우에는 양절메밀에서는 발아 후 12 시간이내에 증가되었으나, 대관 3-3호에서는 36시간까지 유전자 발현양 증가하는 것을 확인할 수 있었다. 이러한 제주산 메밀의 발아초기 단계별 전사체 분석 데이터는 종간의 기능적, 형태학적 차이를 일으키는 메커니즘 규명에 도움을 줄 것으로 사료된다.