Earticle

The aim of this study was to identify allele variability and frequencies of six microsatellite markers (BM861, INRA124, INRA189, UMN0103, UMN0307 and UMN0504) specific to the Y-chromosome. DNA samples from 147 males of three Korean native and seven exotic cattle breeds were tested. Three (Chikso, Heugu, and Jeju black cattle) Korean native cattle (KNC) breeds showed Bos taurus genotype. In the neighbor-joining tree, based on Nei’s DA genetic distance, ten breeds represented main group. In addition, Bayesian clustering result showed that 3 main clusters are for individual allele variability and frequencies. Moreover, KNC breeds showed differences in genotype with B. taurus type when compared to previous studies. The molecular information of paternal lineage in this study would be useful for the conservation and utilization of three KNC breeds as genetic resources.

Simple sequence repeats (SSRs) were isolated from major Pleurotus pulmonarius cultivars in Korea, namely ‘HS47’ (monokaryon, gamete of ‘Santari’), ‘GB19’ (monokaryon, gamete of ‘Santari’), ‘Hosan,’ ‘Yeoleumneutali1,’ ‘Sambok,’ ‘Gangsan,’ ‘Yaksan,’ ‘Jasan,’ ‘Hyangsan,’ and ‘Yeoleumneutali2,’ and characterized via HiSeq genome sequencing and bioinformatic analysis. The genome sizes of the monokaryons ‘HS47’ and ‘GB19’ were estimated to be 37.3 and 37.2 Mb, respectively, and those of the other dikaryotic cultivars ranged from 47.1 to 61.1 Mb. A total of 711 (smallest) and 1,106 (1.5 times the smallest) SSRs were found in the ‘HS47’ and ‘Gangsan’ genomes, respectively. Hexanucleotide and octanucleotide motifs accounted for the top two fractions of all SSRs. CGA/TCG, A/T, and CTC/GAG were the most frequently detected nucleotides in the SSRs. Most of the SSRs were 21~30 nucleotides long (hypervariable for application), accounting for 70% of all lengths of SSRs.

To develop a method for the differentiation of Pleurotus ostratus cultivars, the multiplex-simple sequence repeat (SSR) primer set based on the SSRs obtained from whole genomic DNA sequence analysis was designed with two polymerase chain reaction (PCR) primer sets. These SSR primer sets were employed to distinguish 10 cultivars and strains. Twenty polymorphic markers were selected based on the genotyping results. PCR with each primer produced 1–4 distinct bands ranging in size from 150 to 350 bp, which was within the expected range. However, since a sole SSR marker was unable to detect polymorphisms in every cultivar, multiplex PCRs with composite PCR primer sets were employed. The multiplex primer, “166+115,” completely discriminated 12 cultivars and strains with 40 loci, which were 12 more than the simple arithmetic addition of each locus of the primers 115 and 166. These results might be useful to provide an efficient method for the differentiation of P. ostreatus cultivars with separate PCRs for the quality control of spawn and protection of breeders’ rights.

한 개체군의 초기 점유 및 후기 분화 과정들은 개체군의 유전적 다양성과 유전적 분화구조에 기록되며, 외래종들의 사례는 이러한 개체군유전학 연구들을 수행할 좋은 기회를 제공한다. 본 연구에서는 한·중·일 내 12개체군에서 채집된 도마뱀부치를 대상으로 mtDNA와 microsatellite 마커를 이용하여, 개체군들의 유전적 다양성을 평가하고, 이를 바탕으로 개체군 간 유전적 구조 및 확산의 역사적 과정을 파악해보고자 하였다. mtDNA 자료의 분석결과 한·중·일 간에는 유전적으로 분화된 상태에서 다른 도마뱀들에서 알려진 것보다 30%이상 낮은 유전적 다양성을 가지고 개체군들이 정학한 것으로 확인되었으 나, 개체군 간 유전적 분화정도와 지리적 거리와는 유의미한 관련성이 없었다. 반면, microsatellite 결과의 경우, 한·중·일 간 유전적 분화수준에서 차이가 없고, 예측되는 이형접합 체의 비율(HE)은 기존의 알려진 유사종들에 비하여 크게 낮았으며, 대부분의 개체군 간에는 유 의한 유전적 분화상태를 보여주었다. 그러나 역시 개체군 간 유전적 분화정도와 지리적 거리와 는 유의미한 관련성이 없었다. 이러한 연구결과들을 종합할 때, 도마뱀부치 개체군들은 자연분산보다는 인위적으로 중국, 일 본, 한국 순으로 유입되었을 가능성을 보여준다. 정착 이후 개체군들 간 교류로 인하여 유전적 구조가 유사해 졌지만, 각 개체군의 유전다양성은 여전히 낮아 잠재적인 서식지 훼손이나 개체 군의 교란에 취약할 것으로 판단되었다. 본 연구는 교육부의 재원으로 한국연구재단의 지원을 받아 수행된 기초연구사업 (2016R1D1A1B03931085)연구입니다.

The aim of this study was to provide basic guidelines for conservation and management of endangered plants in the national parks of Korea. Pedicularis hallaisanensis Hurus (Scrophulariaceae) is considered a second-class endangered species by Korean government and it is listed as a EN (Endangered) species in Red Data Book of Korea. We analyzed ecological conditions of P.hallaisanensis habitats based on vegetation properties and soil characteristics. This species which is known to inhabit in sub-alpine grassland of Korea and its population was located at an elevation of 1,374m to 1,916m. In the study sites, soil organic matter, total nitrogen, soil moisture, available phosphate, exchangeable potassium, exchangeable calcium, exchangeable magnesium, exchangeable sodium, cation exchange capacity and soil pH were 2.55-22.48%, 0.10-0.71%, 17.20-57.57%, 1.98-7.77ppm, 0.05– 0.22cmol+/kg, 0.78-18.21cmol+/kg, 0.25-0.76cmol+/kg, 0.08-0.13cmol+/kg, 5.35-26.48cmol+/kg and 5.69-7.55 respectively. Additionally, the genetic variation and structure of three populations were assessed using microsatellite markers.The genetic diversity of P.hallaisanensis among populations was the highest in Mt. Gayasan. Analysis of molecular variance (AMOVA) apportioned among population and within population variations, showing more genetic variance (52%) occurred in within population, and 48% variation among populations. Lastly, we developed predicted distribution model based on climate and topographic factors by applying SDMs (Species Distribution Models). Consequently, current status of P. hallaisanensis habitats were very fragile and vulnerable with the pressure of the vegetation and the climate change constantly threatening the species’ survival. Therefore, it is essential to establish in situ conservation strategies for protecting natural habitats and to require exploring potential and alternative habitats for reintroduction.

Molecular genetic markers were genotyped used to detect chromosomal regions which contain economically important traits such as growth traits in pigs. Three generation resource population was constructed from a cross between the Korean native boars and Landrace sows. A total of 193 F2 animals from intercross of F1 were produced. Phenotypic data on 7 traits, birth weight, body weight at 3, 5, 12, 30 weeks of age, live empty weight were collected for F2 animals. Animals including grandparents (F0), parents (F1), offspring (F2) were genotyped for 194 microsatellite markers covering from chromosome 1 to 18. Quantitative trait locus analyses were performed using interval mapping by regression under line-cross model. To characterize presence of imprinting, genetic full model in which dominance, additive and imprinting effect were included was fitted in this analysis. Significance thresholds were determined by permutation test. Using imprinting full model, four QTL with expression of imprinted effect were detected at 5% chromosome-wide significance level for growth traits on chromosome 1, 5, 7, 13, 14, and 16.

본 연구는 버크셔종의 육질에 대한 특성을 분석하여 국내 돈육시장에서의 적합성을 확인하고, MS marker를 이용한 육질 우수 개체 선발의 효율성에 대해서 분석하였다. 버크셔 323두를 동일 사양조건에서 사육하고 도축하여 육질형질을 분석하고, 혈액으로부터 genomic DNA를 분리하여 50개의 MS marker에 대한 유전자형을 분석하였다. 버크셔종은 국내에서 가장 많이 이용되고 있는 듀록, 요크셔, 랜드레이스 종보다도 육질에 있어서 더욱 우수한 특성을 나타내었다. 특히 도축 24시간 후 pH 값은 평균 5.88±0.01로 대단히 높게 나타났고, 지방함량의 경우에도 2.878±0.06로 확인되었다. MS marker와의 연관성을 확인한 결과, 14개 MS marker가 육질형질과 연관성을 있는 것으로 나타났다(p<0.05). 특히 pH24hr와 지방함량에서는 가장 높은 값을 가지는 대립유전자집단이 가장 낮은 집단에 비해 최고 0.55, 2.04%가 높은 것을 확인할 수 있었다. 이러한 특성을 버크셔종의 육질 우수 개체 선발에 이용한다면 육색, 지방함량, pH24hr, 가열감량, 드립감량, 그리고 보수성에서 육질 형질값의 높은 개량효과를 기대할 수 있을 것으로 판단된다.

In this study, the efficiency of microsatellite(MS) markers for pork quality was examined and further, their suitability to domestic pork industry also was verified, by measuring meat quality parameters of Berkshire breeds. A total of 323 pigs of Berkshire breeds were slaughtered and subjected to meat quality evaluation. In addition, the genomic DNAs from blood samples of slaughtered pigs were used for genotyping analysis of 50 MS markers. The results revealed that Berkshire breeds have excellent meat quality, compared with the popular domestic breeds such as Duroc, Yorkshire, and Landrace. Noticeably, the Berkshire breeds exhibited a significant post-mortem pH24hr(5.88±0.01) and fat content(2.878±0.06). Through the linkage analysis between MS markers, 14 MS markers showed significant association with meat quality traits(p<0.05). Maximum significant differences of 0.55 pH24hr value and 2.04% fat content were observed between the highest and lowest allele populations. If these 14 MS markers are applied to the pork quality diagnosis kit, the synergistic effect can be expected in meat quality parameters such as meat color, fat content, pH 24hr, cooking loss, drip loss and water-holding capacity.

Genotype data of eleven microsatellites typed in 713 Hanwoo breed(Korean cattle) animals were used to estimate the genetic diversities and relationship of nine brand populations distributed in six regional areas. Size of microsatellite markers decided using GeneMapper Software(v.4.0) after analyze in kinds of ABI machine of name of 3130 using ABI Stock MarkersTM kit. Frequencies of microsatellites markers were used to estimate heterozygosities, gene diversities and genetic distances. Microsatellite markers showed high polymorphism and 163 alleles were detected from the genotyping of eleven microsatellite markers. Allele frequency results were resemblant to each other in case of most markers and this between each brand where several alleles shared the majority mean. Genetic distances between populations were obtained using Nei’s DA distance method. Expected heterozygosity between each brand population was estimated very analogously. Average expected heterozygosity(0.765) of whole Hanwoo populations had higher diversities than other species of Europe, North American continent and spanish breeds. Genetic distances(0.0413) between ChoneBuk(JS) and ChoneNam(SJ) was nearest than distances between other populations. Genetic distances between KyongGi(AS, YP) and KyongNam(NH) showed far distance than other populations by 0.0493 and 0.0489 each respectively. ChoneNam(SJ) showed near genetic distance relatively with other areas' populations. In the UPGMA tree that is made based on DA distance matrix, some brand populations ramified to area. Because this became evenly genetic because gene flow of Hanwoo was frequent comparatively between each area by stock bull selection of country unit and their sperms(KPN) supply system that is done till today. It can be considered that this effect is by transfer of calf between near geographical areas. Each individuals were not ramified to different group and were spread evenly in phylogenetic dendrogram about all Hanwoo of each brand.

한우를 포함한 동북아시아 및 중국 내륙지역의 7개 품종, 유럽 Bos taurus 5개 품종, 아프리카 Bos taurus 2개 품종, 아시아 Bos indicus 2개 품종, 아프리카 Bos indicus 2개 품종 및 참고집단으로 인도네시아 반텡종인 Bali cattle의 19개 소품종을 11개 microsatellite markers를 이용하여 유전적 다양성 및 특성, 유전적 거리 추정 및 분지도 작성 등을 통하여 한우를 비롯한 동북아시아 소 품종들에 대한 보다 더 명확한 유전적 유연관계에 대하여 고찰하고자 하였다. 분석된 11개의 microsatellite loci에서 대립유전자의 수는 9개(ILSTS005 및 TGLA227)에서 26개(TGLA122)까지 검출되었고 전체 대립유전자수는 162개로 충분한 유전적 정보를 제공해 주었다. 또한 대립유전자별 빈도에 있어서 품종별 현저한 차이를 보이는 microsatellite loci들이 다수 검출되었다. 품종별 이형질성에 있어 아시아 품종들이 유럽계 및 아프리카 계통의 taurines 보다 높게 나타났으며, 아시아 및 아프리카 indicines 품종보다는 낮게 분석되어졌다. 유전적 거리 추정결과로 한우를 포함한 동북아시아 품종들은 같은 조상에서 유래된 품종 집단으로 추정되고 일부 품종들은 유럽계 품종들의 유전적 영향을 받은 것으로 나타났으며, 그 중 일본 흑모화우가 가장 많은 영향을 받은 것으로 분석되어졌다. 그리고 동북아시아 품종들과 Bos indicus 품종들 간의 유전적 거리가 유럽계통 및 아프리카 계통의 Bos taurus 품종들과 Bos indicus 품종들과의 거리만큼이나 유전적으로 상이하였으며, 유럽계 소 품종 및 아프리카 taurines 품종들과도 어느 정도의 유전적 거리가 인정되어 한우가 Bos taurus 및 Bos indicus 품종들의 교잡에 의해 형성되지 않았음을 확인할 수 있었으며, 또한 동북아시아 소 품종들은 지역적 별개의 가축화 기원의 가능성이 있음을 제기한다.

For the genetic assessment of the cattle breeds including Hanwoo, eleven microsatellite markers on ten bovine autosomes were genetically characterized for 618 individuals of nineteen cattle breeds; North Eastern Asian breeds (Korean cattle, Korean Black cattle, Japanese Black cattle, Japanese Brown cattle, Yanbian cattle), Chinese yellow cattle (Luxi cattle, Nanyang cattle), European Bos taurus (Angus, Hereford, Charolais, Holstein, Limousin), African Bos taurus (N'Dama, Baoule), African Bos indicus (Kavirondo Zebu, White Fulani), Asian Bos indicus (Sahiwal, Nelore) and one Bali cattle, Bos banteng as an outbreed-reference population. Allele frequencies derived from the genotyping data were used in estimating heterozygosities, gene diversities and genetic distances. The microsatellite loci were highly polymorphic, with a total of 162 different alleles observed across all loci. Variability in allele numbers and frequencies was observed among the breeds. The average expected heterozygosity of North Eastern Asian breeds was higher than those of European and African taurines, but lower than those of Asian and African indicines. Genetic distances were estimated using Nei's DA genetic distance and the resultant DA matrix was used in the construction of the phylogenetic trees. The genetic distances between North Eastern Asian cattle breeds and Bos indicus were similar with those between European Bos taurus and Bos indicus, and African Bos taurus and Bos indicus, respectively. The clusters were clearly classified into North Eastern Asian, European and African taurines groups as well as different cluster with Chinese mainland breeds, firstly out-grouping with Bos indicus. These results suggest that Korean cattle, Hanwoo, had not been originated from a crossbred between Bos primigenius in Europe and Bos indicus in India and North Eastern Asian Bos taurus may be have separate domestication from European and African Bos taurus.

The objective of present study was to ascertain parentage of Thoroughbred(TB) horses in Korea. A total of 2,029 TB horse samples including 993 foal samples for parentage testing were genotyped for nine international minimum standard markers(AHT4, 5, ASB2, HMS3, 6, 7, HTG4, 10, and VHL20). This method consisted of multiplexing PCR procedure, and showed reasonable amplification of all PCR products. Genotyping was performed with an ABI 310 genetic analyzer. The number of alleles per locus varied from 5 to 11 with a mean value of 7.33 in TB. Expected heterozygosity was ranged from 0.544 to 0.837(mean 0.709) and the total exclusion probability of 9 microsatellites loci was 0.9978. Of the 9 markers, ASB2, HMS7 and HTG10 loci have relatively high PIC value( > 0.7). All of the 993 foals were qualified by compatibility according to Mendelian fashion in the present DNA typing for parentage testing. These results suggest that the present DNA typing has high potential for parentage verification of TB horses.

돼지 6번 염색체에서 근내지방 함량과 등지방 두께와 관련된 QTL이 탐색되어진 영역(6q28- 6q32)에서 미세지도 작성을 위한 유용한 marker를 개발하기 위하여 실시하였다. 대량 염기서열 분석 자료를 근거로, 반복염기서열 분석을 수행한 결과 KP0290F2(TTCC), KP0248C11(AAAT), KP1231C91 (TAG), KP1231C92(TTG) 그리고 KP1231C93(GA)의 5 부위에서 다형성을 나타내었다. 이 부위들에 대한 랜드레이스, 재래돼지, 듀록, 요크셔, 버크셔, 오지산돈, 향돈 그리고 민돈의 8품종에 대한 유전자형 분석 결과 평균 대립유전자의 수는 2.13, 4.63, 7.38, 2.75 그리고 6.25로 나타났다. 그리고 8 품종에 대한 KP0290F2, KP0248C11, KP1231C91, KP1231C92 그리고 KP1231C93에 대한 평균 heterozygosity 값을 산출한 결과, 0.2110, 0.6865, 0.8304, 0.4057 그리고 0.7051로 나타났으며, 5 markers에 대한 8 품종의 평균 heterozygosity 값은 0.6313, 0.5662, 0.5814, 0.6957, 0.4517, 0.4847, 0.5758 그리고 0.5559로 나타났다. KP0248C11, KP1231C91 그리고 KP1231C93은 적절한 대립유전자 수를 나타내었고, 또한 hetetozygosity 값이 높게 나타났을 뿐만 아니라, 표준편차도 적게 나타난 점으로 미루어 보아 앞으로 유용한 marker로서 이용이 가능할 것이라 사료된다. 본 연구의 결과 개발된 marker는 SW71(98.6 cM)과 SW1881(121.1 cM) 영역내에 존재하는 유용한 유전자를 발굴하기 위한 미세지도 작성에 유용하게 활용될 수 있는 marker로 판단되며, positional cloning에도 이용 할 수 있을 것이라 사료된다. 또한 돼지 게놈 연구가 완성되어 염기배열이 밝혀지기 전에 이용 가능한 표지인자들을 다량으로 확보 할 수 있을 것이라 사료된다.

This study was conducted to develop new markers at the region that was related to QTL affecting intramuscular fat and backfat thickness on chromosome 6q28-6q32 in pigs. Dozens of repeated sequences were founded using shotgun sequencing of several BAC clones corresponding to that region, of which five new microstellite markers that identified polymorphism were discovered. The mean number of alleles at each locus observed 2.13(KP0290F2), 4.63(KP0248C11), 7.38(KP1231C91), 2.75(KP1231C92) and 6.25(KP1231C93) in 8 breeds(Landrace, Korean native pig, Duroc, Yorkshire, Berkshire, Wuzhishan pig, Xiang pig, Min pig). The average estimated heterozygosity values at each locus varied from 0.2100(KP0290F2) to 0.8304(KP1231C91) in all populations. In other hand, the average allele of all loci was within range of 0.4517(Berkshire) and 0.6957 (Yorkshire). Of these markers, KP0248C11, KP1231C91 and KP1231C93 were identified to have optimal number of alleles, high heterozygosity values and low standard deviation values. Especially, KP1231C91 and KP1231C93 might be considered as a useful marker for genetic mapping and diversity study.

국내에서 사육중인 치와와 31두, 풍산개 20두, 래브라도 리트리버 8두를 대상으로 micro- satellite DNA형의 유전자 빈도에 기초하여 heterozygosity, PIC, 그리고 PE를 분석한 결과를 요약하면 다음과 같다. 치와와의 대립유전자 수는 4∼14개로서 expected heterozygosity와 PIC는 각각 0.432∼0.883 (평균 0.711), 0.397∼0.856(평균 0.659)으로 나타났으며 PEZ1, PEZ3, PEZ6, PEZ10, PEZ12의 marker는 PIC 0.7이상으로 나타났고 14개 marker를 조합시 부권부정율은 0.9999로 관찰되었다. 풍산개의 대립유전자 수는 2∼9개로서 expected heterozygosity와 PIC는 각각 0.262∼0.817 (평균 0.559), 0.222∼0.772 (평균 0.503)으로 나타났고 PEZ1, PEZ6, PEZ13의 marker는 PIC 0.7이상으로서 16개 marker를 조합시 부권부정율은 0.9991로 관찰되었다. 래버라도 리트리버의 대립유전자 수는 3∼5개로서 expected heterozygosity와 PIC는 각각 0.425∼0.808 (평균 0.660), 0.354∼0.717 (평균 0.563)으로 나타났고 PEZ8, PEZ12의 marker는 PIC 0.7이상으로 관찰되었으며 12개 marker를 조합시 부권부정율은 0.9968로 관찰되었다. 이상의 결과는 microsatellite DNA형에 의한 개의 친자감정 및 개체식별에 유용한 자료로 활용할 수 있을 것으로 사료된다.

This study was carried out to investigate a usefulness of the microsatellite DNA markers for individual identification and parentage verification in three dog breeds. A total of 59 random dog (31 Chiwawa, 20 Poongsan, 8 Labrador Retriever) samples were genotyped by using 14 markers (Chiwawa dog), 16 markers (Poongsan dog), and 12 markers (Labrador Retriever dog) among the 17 international standard markers (PEZ1, 3, 5, 6, 8, 10, 11, 12, 13, 15, 16, 17, 20, 21, FHC2010, FHC2054 and FHC2079), respectively. The number of alleles per locus varied from 4 to 14 with a mean value of 6.07 in Chiwawa dog, 2 to 9 with a mean of 4.75 in Poongsan dog, and 3 to 5 with a mean of 4.00 in Labrador Retriever dog. Observed heterozygosity was ranged 0.419∼0.968 (mean 0.755), 0.300∼0.950 (mean 0.597) and 0.125∼0.750 (mean 0.604), and expected heterozygosity was ranged 0.432∼0.883 (mean 0.711), 0.262∼0.817 (mean 0.559) and 0.425∼0.808 (mean 0.660) in these three dog breeds. PIC value was ranged 0.397∼0.856 (mean 0.659), 0.222∼0.772 (mean 0.503) and 0.354∼0.717 (mean 0.563) in these three dog breeds. Of the 17 markers, PEZ1, PEZ3, PEZ6, PEZ10, PEZ12 loci, PEZ1, PEZ6, PEZ13 loci, and PEZ8, PEZ12 loci have relatively high PIC value (＞0.7) in Chiwawa dog, Poongsan dog and Labrador Retriever dog, respectively. The exclusion probability was ranged 0.240∼0.741, 0.111∼0.616, and 0.198∼0.529, and the combination of microsatellite loci was 0.9999, 0.9991, and 0.9968 in Chiwawa dog, Poongsan dog and Labrador Retriever dog, respectively. These results can give basic information for developing parentage verification and individual identification system in these three dog breeds.

돼지 원산지 추적 및 브랜드육 식별을 위한 MS marker set를 확립하기 위해 EU의 EID+DNA DNA Tracing 연구 project, 국제동물유전학회 및 Roslin 연구소 등에서 제안한 돼지의 다형성 분석에 사용되는 MS marker 중 17종을 선발하여 다형성지수, F-통계량, 동일개체 출현확률 및 친자감별확률 등을 MSA, CERVUS, FSTAT, GENEPOP 및 API-CALC 프로그램 등을 연계적으로 활용하여 추정하였다. 다형성지수 추정치를 1차 선발요인으로 하고, 각 MS marker의 PCR 결과물의 크기 및 분석 비용의 경제성을 추가로 감안하여 최종 13종의 MS marker(SW936, SW951, SW787, S00090, S0026, SW122, SW857, S0005, SW72, S0155, S0225, SW24 and SW632)와 성감별을 위한 2개의 성감별 primer로 조합된 하나의 Multiplexing PCR 시스템을 확립하였다. 이를 이용해 돼지 사육환경을 무작위 교배집단(PI), 반형매 교배집단 그리고 전형매 교배집단으로 가정하여 동일개체 출현확률을 분석한 결과 2.47×10－18, 6.39×10－13 Table2.Expected probability of identity(PI), among genotypes of random individuals, probability of identity among genotypes from random half sibs(PIhalf-sibs), among genotypes of random individuals, probability of identity among genotypes from random sibs(PIsibs), exclusionary power of the second parent(PE), and the probability of parent exclusion when both parents are unconfirmed(PEpu) No. of MS PI PIhalf-sibs PIsibs PEpu PE 17a 1.09×10-17 1.09×10-13 1.09×10-7 0.99974 1.00000 13b 2.47×10-18 6.39×10-13 1.08×10-8 0.999998 0.999669 a, the 17 MS markers in Table 1 and b, the 13 MS markers (SW936, SW951, SW787, S00090, S0026, SW122, SW857, S0005, SW72, S0155, S0225, SW24 and SW632) selected from the 17 MS markers by the rank of heterozygosity and PCR product size 그리고 1.08×10－8로 나타나 브랜드 식별 및 원산지 추적에 적합할 것으로 사료된다.

Bovine microsatellite 마커인 INRA124는 소에 있어 Y 염색체 특이 마커로서 130 및 132 bp의 두 개 대립유전자를 가지고 있는데, Bos taurus 특이 대립유전자는 132 bp이고, Bos indicus 특이대립유전자는 130 bp이다. 이번 연구는 이 유전좌위를 이용하여 한우집단에 대한 Bos taurus 및 Bos indicus 유래의 대립유전자 분석을 통하여 한우의 유전적 특성을 이해하고자 하였다. 동북아시아, 중국내륙, 유럽, 인도 및 아프리카 유래의 20개 소 품종 822두의 수컷을 공시하여 분석해 본 결과, 유럽계통, 일본 화우 및 한우에서는 Bos indicus 유래의 대립유전자는 전혀 검출되지 않았으며, 인도 및 아프리카 유래의 Bos indicus 품종에서는 132 bp의 대립유전자가 전혀 검출되지 않았다. 한우, 브라만 및 샤롤레의 교잡우 집단(CBK)에서는 Bos indicus 특이 대립유전자인 130 bp가 0.19의 빈도로 검출되었고, 중국내륙 품종인 노서우 및 난양우는 각각 0.46 및 0.29의 빈도로 검출되었다. 이로서 한우의 기원은 기존 혼합설과 달리 유럽계통의 Bos taurus만으로 기원하였을 가능성을 제기한다.

The INRA124 is a bovine Y-chromosomal specific microsatellite locus that has been revealed a polymorphism. This locus has two alleles. The 132 bp allele is specific to cattle (humpless) of taurine origin and the 130 bp allele is specific to cattle (humped) of indicine origin. A total 822 males of 20 breeds or populations; North Eastern Asian breeds (Hanwoo, Korean Black cattle, Chik-so, CBK, Japanese Black cattle, Japanese Brown cattle, Yanbian cattle), Chinese yellow cattle (Luxi cattle, Nanyang cattle), European origin (Angus, Hereford, Charolais, Simmental, Brown swiss, Holstein, Limousin), African origin (Kavirondo zebu, White Fulani, crossbreed of N'Dama and Boran), Indian origin (Sahiwal) were characterized the distribution of alleles using INRA124 locus. Any individuals of European, Japanese origins and Hanwoo were not detected 130 bp allele, Bos indicus specific allele. Bos indicus breeds of Indian and African origins were not detected 132 bp allele, Bos taurus specific allele. CBK population that the crossbreed of Hanwoo, Brahman and Charolais showed the frequency of 0.19 in indicine specific allele. The breeds of Chinese mainland, Luxi and Nanyang cattle were detected 0.46 and 0.29 frequencies in indicine specific allele, respectively. These results suggest that Korean cattle, Hanwoo, had not been originated from a crossbred between Bos primigenius in Europe and Bos indicus in India.

14개의 microsatellite(MS) marker를 사용 할 경우 무작위 교배 집단(PI) 가정 하에 3.43×10－27의 판별율을 보여 60개의 single nucleotide polymorphism(SNP) marker에 비해 약 1,000배의 높은 판별 효과를 나타내는 것으로 파악되었다. 그러나, 60개의 SNP marker의 경우 반형매 교배 집단(PIhalf-sibs)으로 가정할 경우 4.69×10－20과 전형매 교배 집단(PIsibs)으로 가정 할 경우 8.02×10－12으로 14개의 MS marker에 비해 약 10배와 10,000배의 높은 판별 효과를 나타내는 것으로 추정되었다. 이러한 결과는 무작위 교배집단에서는 사용된 marker의 전체 대립유전자수(MS:SNP = 146:120)에 의하여 판별효율이 결정되는 반면, 혈연관계가 높은 반형매와 전형매 집단에서는 비슷한 총 대립유전자수일 경우 marker의 수(MS:SNP=14:60)가 많은 경우가 더 높은 판별율을 보이는 것으로 나타났다. 한육우의 경우 소수의 보증 종모우를 이용해 인공수정을 통해 형성된 거대한 반형매 집단으로 가정하였을 경우 MS와 SNP marker의 판별율은 10배 정도의 차이로 큰 차이를 보이지 않을 것으로 예견되나, likelihood rato를 이용 하는 inclusion 방법에 의하여 부모를 동시에 찾을 확률은 MS marker가 1,000 배 정도 더 효율적인 것으로 나타났다. SNP marker의 장점인 변이의 안정성, 유전자형 분석의 자동화 및 대용량화 등을 한육우의 동일성 검사에 활용하기 위해서는 분석비용 절감 방안과 분석방법 및 장비의 국산화 등 실용 및 상용화적 측면에서의 연구개발이 필요하다고 사료된다.

When 14 microsatellite(MS) markers were applied in the identifying test for 480 Hanwoo, the discriminating power was estimated as 3.43×10－27 based on the assumption of a random mating group(PI). This rate is 1,000 times higher than that of 60 single nucleotide polymorphism(SNP) markers. On the other hand, the power of the 60 SNP markers was estimated as 4.69×10－20 and 8.02×10－12 on the assumption of a half-sib mating group(PIhalf-sibs) and a full-sib mating group(PIsibs), respectively. These powers were 10 times and 10,000 times higher than those of the 14 MS markers. The results indicated that the total number of alleles(MS vs SNP = 146 vs 120) acted as a key factor for the discriminating power in a random mating population, and the total number of markers(MS vs SNP = 14 vs 60) was a dominant influence on the power in half-sib and full-sib populations. In the Hanwoo population, in which it was assumed that the entire population is the enormous half-sib group formed by the absolute genetic contribution of a few nuclear bulls, there will be only a 10 times difference in the discriminating power between the 14 MS markers and the 60 SNP makers. However, the probability of not excluding a candidate parent pair from the parentage of an arbitrary offspring, given that only the genotype of the offspring(PNEpp) was 1,000 times higher as shown by the 14 MS markers than that by the 60 SNP markers. The strong points of SNP makers are the stability of the variation(low mutation rate) and automation of high-throughput genotyping. In order to apply these merits for the practical and constant Hanwoo identity test, research and development are required to set a cost-effective platform and produce a homemade apparatus for SNP genotyping.

본 연구에서는 Landrace, Large White와 Duroc 3품종의 3원 교잡 시스템으로 비육돈을 생산하는 9개 농가를 대상으로 Landrace와 Large White 교배를 통해 생산된 F1 모돈과 Duroc 웅돈 그리고 비육돈을 이용하여 기 보고한 바 있는 돼지 이력추적 및 브랜드육 식별을 위한 13종의 MS marker의 개체판별능과 혈연관계 추정 효율을 실증하였다. 우선 F_1 모돈과 웅돈 즉 부모 집단을 대상으로 API-CALC version 1.0 프로그램을 이용하여 무작위 교배집단, 반형매 교배집단 그리고 전형매 교배집단으로 가정 시 개체 판별능이 각 각 4.94×10^(－34), 8.16×10^(－23) 그리고 2.01× 10^(－08)으로 추정되었으며, 비육돈을 포함한 추정치는 3.74×10^(－35), 5.48×10^(－25) 그리고 2.96×10^(－11)으로 추정되었다. 또한 Cervus version 2.0을 이용하여 친자감별률을 추정한 결과 100% 인 것으로 추정되었다. 이론적으로 산출된 상기의 수치들을 검증하기 위해 PAPA version 2.0 프로그램을 이용하여 비육돈 전체 452두에 대한 친자감별을 실시한 결과 100% 친부모를 확인 할 수 있었으며, Cervus 프로그램을 이용하여 전체 축군에서 동일한 대립유전자형을 가진 개체의 출현을 조사한 결과 동일개체는 존재하지 않는 것을 확인하였다. 비록 개체 판별능에 대한 실증은 제시한 이론적 판별능에 상응하는 두수를 검증하지는 못하였으나, 친자확인의 경우는 제시한 이론적 효율성 100%는 실증적으로 검증되었다. 따라서 현재 국내에서 행해지는 3 품종 교잡에 의한 비육돈 생산 시스템의 경우 본 연구의 결과로 비추어 볼 때 13 MS marker를 이용하여 체계화된 전산정보와 병행하여 계열화된 생산체계하의 브랜드 단위 또는 지역별 권역으로 구분하는 이력추적이 충분히 가능하다고 사료된다.

There are four (yellow, brindle, black, black in Jeju) kinds of native cattle in Korea. But only twelve hundred Korean brindle cattle (KBC) are remaining in limited areas of Korea and the genetic lineage, diversity, polymorphisms of KBC has not been identified. To analysis genetic polymorphism of KBC, 33 KBC were characterized using 11 microsatellite markers. Size of microsatellite marker was decided using Gene Mapper software after analysis ABI 3130XL. The average of allele numbers of KBC was 6.7 in this study, but that of Hanwoo was 10.0 in our previous report. The observed and expected heterozygosities of KBC were 0.719 and 0.738 but those of Hanwoo were 0.751 and 0.760 in our previous report. Also polymorphism information content (PIC) values were average 0.690 in KBC but 0.725 in Hanwoo. These results demonstrate that genetic polymorphism of KBC have decreased because the population was limited.

In this study, genotyping was executed by using 11 microsatellite markers (BM1824, BM2113, ETH10, ETH225, ETH3, INRA23, SPS115, TGLA122, TGLA227, TGLA53, and TGLA126) for diversity of 214 Hanwoo cows in Hoengseong area. Each marker's size and number of allele, observed heterozygosity, expected total heterozygosity, and polymorphism information content were analyzed by 11 Microsatellite marker. The average of size range was detected from 150.9 to 174.9 in Hanwoo cows of Hoengseong. The number of average allele was 10.0 that is similar to the average of Kangwon Hanwoo, which is known as 10.5 in the previous report. The average were 0.751 for observed heterozygosity, 0.760 for expected total heterozygosity, 0.725 for polymorphism information content, respectively. These results were similar to previous studies in Kangwon Hanwoo, National Hanwoo and Korean Proven Bulls. This study is expected to contribute for genetic improvement of Hanwoo cows in Hoengseong as a popular brand.