This study was carried out to examine a molecular marker system for parentage test in Jeju Black cattle (JBC). Based on the preliminarily studies, we finally selected for construction of a novel genetic marker system for molecular traceability, identity test, breed certification, and parentage test in JBC and its related industrial populations. The genetic marker system had eight MS markers, five indel markers, and two single nucleotide polymorphisms (SNPs; g.G299T and g.del310G) within MC1R gene which is critical to verify the breed specific genotypes for coat color of JBC differing from those of exotic black cattle breeds such as Holstein and Angus. The results showed lower level of a combined non-exclusion probability for second parent (NE-P2) of 4.1202×10-4 than those previously recommended by International Society of Animal Genetics (ISAG) of 5.000×10-4 for parentage, and a combined non-exclusion probability for sib identity (NE-SI) of 2.679×10-5. Parentage analysis has been successfully identified the JBC offspring in the indigenous population and cattle farms used the certified AI semens for production using the JBC-derived offspring for commercial beef. This combined molecular marker system will be helpful to supply genetic information for parentage test and traceability and to develop the molecular breeding system for improvement of animal productivity in JBC population.

This study was conducted to find a useful marker for gene polymorphism analysis using Microsatellite marker (MS marker) in Gyeongju Donggyeong dog. Twenty three MS marker analyzed the genetic features of DNA using 100 Gyeongju Donggyeong dogs in Gyeongju area. It was performed multiplex PCR with 3 set primer divided 9, 10 and 4 by analysis of conditions among MS markers. The results were calculated heterozygosity, polymorphic information content (PIC), allele frequency and number of allele at each locus using Microsatellite Toolkit software and Cervus 3.0 program. Total 148 alleles were genotyped to determine and average 6.43 alleles was detected. FH3381 had the highest of 15 alleles and FH2834 had the lowest of 2 alleles. Expected heterozygosity had a wide range from 0.282 to 0.876 and had average value of 0.6496. Also, Observed heterozygosity had a more wide range from 0.200 to 0.950 and had average value of 0.6404. PIC had range from 0.262 to 0.859 and average PIC was calculated 0.606. Especially, FH2998 represented the highest rate of observed heterozygosity of 0.950 and FH3381 represented the highest rate of expected heterozygosity of 0.876 and PIC of 0.859. The use of these markers was considered to be useful to study genetic traits of Gyeongju Donggyeong dog.

3종의 초위성체 마커를 이용해 분석한 대립유전자형을 F0, F1 그리고 F2로 세대 간 구분 하여 동일한 개체 출현확률 값을 추정한 결과 F2의 무작위교배집단으로 가정한 경우 13종의 초위성체 마커는 3.84 × 10-23의 추정치를 나타내 37개의 SNP 마커를 이용할 경우와 유사한 동일개체 출현확률 추정치를 유추할 수 있었다. 본 연구에 사용한 실험축군은 2품종 상호교배로 만들어졌다. 친자감정확률 추정치를 전체집단을 대상으로 13종의 초위성체 마커와 37개의 SNP 마커를 이용하여 분석한 결과 부모를 동시에 찾을 수 있는 추정치인 PEpu의 경우 초위성체 마커는 0.97897이고 SNP 마커는 0.99149였으며, 한쪽 부모를 찾을 수 있는 추정치인 PE의 경우 초위성체 마커는 0.99916이고 SNP 마커는 0.99949로 나타났다. 또한 가능한 후보 부모들로부터 가장 확률이 높은 부모를 찾을 추정치인 PNEpp의 경우는 초위성체 마커와 SNP 마커 둘 다 1.00000으로 추정 되었다. 한정된 부모집단 내 한정된 대립유전자형을 통해 대량의 비육돈이 생산되는 국내의 양돈산업의 경우 DNA 마커의 특성, 분석집단의 크기, 유전자형 분석의 정확도와 비용, 분석된 자료 관리의 용이성 및 기존 분석 시스템과의 호환성 등을 고려하여 효율성과 경제성이 높은 마커를 선정해 마커 조합을 만드는 것이 필요할 것으로 사료된다.

Honeybees (Apis mellifera) adapted themselves to different geographical and climatical conditions since they had been introduced in Korea. Beekeepers have tried to breed valuable lineages with artificial insemination or conventional mating techniques. However, evaluation of breeding resultants still relies on timeconsuming observation data. Genetical characterization of breeds has proven its usefulness to preserve genetic resources of livestock. In recent years, microsatellites are most commonly used to evaluate population structures and diversities of living organisms in that the characteristics of locus specificity, rich polymorphism, abundant and random distribution over the genome, and their co-dominant inheritance. Determining classic genetic distances using neutral, highly polymorphic microsatellite markers is a reliable method to investigate genetic relationships and breed differentiation. This methodology can be used to establish preservation priorities for livestock breeds. The final aim of this study was to develop potent markers for assessing genetic structure of lineages after artificial insemination. In this study, the genetic structure of ten microsatellite markers were sequenced or analysed with polymerase chain reaction for eleven European honeybee populations. The results may help to develop reliable microsatellite markers for more efficient preservation strategies of valuable honeybee breeds.

We developed and characterized five polymorphic microsatellites of Nilaparvata lugens from hybridization method using biotin enrichment strategy and two polymorphic microsatellites from Next Generation Sequencing. Also 11 microsatellites that developed by Sun et al. (2011) are employed to carry out genetic analysis of N. lugens in Southeast Asia. The number of alleles per locus ranged from 2 to 12 with an average of 4.63 alleles per locus. The mean observed heterozygosity of the eleven populations ranged from 0.031 to 0.938 and the expected hetetrozygosity ranged from 0.031 to 0.881. Signifiant genetic differentiation was detected among the three N. lugens populations as the FST ranged from 0.028 (Cheong Do and Ha Long) to 0.161 (CH and BN). The results of microsatellite marker suggested that found N. lugens migrated to Korea at least two times in different period or once. Genetic distance of N. lugens between Korea and Hi Pong were mostly closed and genetic distance of Ha Long and HCM were relatively closed. In this study, development of microsatellites should facilitate the study of future population genetics of N. lugens, and eventually elucidate the route of N. lugens migration to Korea. Thus, combining satisfactory microsatellite markers and intensive surveillance methods in paddy field could be easy to understand to the N. lugens migration mechanism.

The bovine fatty acid binding protein 4 and 5 (FABP4 and 5) is a major positional and physiological candidate gene for the bovine marbling and carcass weight. The aim of this study was to evaluate the association between economic traits of Korean cattle (Hanwoo) and genetic variation in fatty acid binding protein 4 and 5 (FABP4 and 5) genes within carcass/meat quality traits and the before/after of fatting in breed Hanwoo. Here, we characterized the nucleotide polymorphism of FABP4 and 5 in 86 cattle. We were detected the variability of three types (GG, AG, and AA) by PCR, and economic traits were analyzed by the mixed regression model implemented in the ASReml program. As the result of statistical and supersonic analysis, FABP4 gene was highly showed significant effect (p<0.006) on marbling score (MS), in contrast FABP5 gene was lowed (p<0.084) on MS before fatting. But, FABP4 gene was highly showed significant effect (p<0.0054) on MS, in contrast FABP5 gene lowest (p<0.0899) on MS in the after of fatting. Compare to supersonic result before fatting in FABP4 gene, it was detected type GG: (p<7.18), AG: (p<8.50), and AA: (p<10.50) (n=50), showed type GG: (p<4.88), AG: (p<2.33), and AA: (p<0.00) after weed out (n=20). Futhermore, it was detected type GG: (p<9.30), AG: (p<7.95), and AA: (p<7.40) (n=50) before fatting in the FABP5 gene. It was shown type GG: (p<2.67), AG: (p<3.50), and AA: (p<5.00) after weed out (n=50). Our results indicate that FABP4 and 5 gene transcription is regulated by the environment of feeding and management, and suggest that feeding and management could be potential key in determining FABP4 and 5 genes transcription for carcass/meat quality traits in breed Hanwoo.

The number of abandoned dogs is increasing with the worsening of the economy and the rising of feed value. It was becoming a serious social problem because of the disease transmission and destruction of natural ecosystems by abandoned dogs been wild animal. In order to solve these problems, companion dogs necessary to secure its own genetic information and to establish the systematic tracking system. Using multiplex-PCR method with 27 microsatellite marker (MS marker) divided 3 set, various alleles occurring to 6 dog breed (Labrador Retriever, German Shepherd, English Springer Spaniel, Belgian Malinois, Jindo Dog, PoongSan Dog) make use of markers to determine allele frequency and heterozygosity. MS marker FH2834 and FH2790 have only two allele and most were found in 13 alleles at FH3381 and FH3399. Average heterozygosity of MS marker is 0.534 and especially, heterozygosity represented the highest value of 0.765 at FH3381. So, it was recognized appropriate allele frequency for individual identification and paternity diagnosis in companion dogs. Using multiplex-PCR method with MS marker, various alleles occurring to dog breed make use of markers to deter mine individual identification and paternity diagnosis, traits associated biomarkers and breed-specific marker for faster, more accurate and ways to reduce the analysis cost. Based on this result, a scientific basis was established to the existing pedigree data by applying genetics additionally. Animal registration system is expected to be conducted nationwide in future. The method expects to very useful this system.

The seven-spotted lady beetle, Coccinella septempunctata (Coleoptera: Coccinellidae) has a broad ecological range, living almost anywhere there are aphids for feeding. In this study, we isolated and characterized a total of 10 microsatellite loci from the species. The loci were validated and characterized using 25 samples collected from five Korean localities. The number of alleles and heterozygosity observed at each locus ranged from 4 to 16 and from 0.37 to 0.89, respectively. None of the loci deviated significantly from Hardy-Weinberg equilibrium and there was no indication of significant linkage disequilibrium among pairs of loci. These microsatellite markers should be very valuable markers for population genetic studies of Coccinella septempunctata.

Lycorma delicatula (White 1845), which has been recently introduced into Korea, is a notorious pest on grapes. This invasive insect has rapidly spread throughout central and southern Korea. To date, we have no behavioral or population genetics information, such as invasion routes and subsequent dispersal rates in Korea, to help understand and control populations of L. delicatula. Here, we have developed 15 novel microsatellite loci for L. delicatula. The isolated loci were polymorphic, with 2 to 19 alleles in 42 individuals from a single population in Cheonan. The analyses revealed that all 42 individuals had different multilocus genotypes with heterozygosity ranging from 0.214 to 0.866. Eleven of the 15 loci did not deviate significantly from Hardy-Weinberg equilibrium. The isolated markers will facilitate population genetic studies of L. delicatula.

Ten novel microsatellite loci were isolated and characterized from the beet armyworm, Spodoptera exiqua. The isolated loci are polymorphic, with 2~12 alleles in 18 individuals from several populations in Korea. All 18 individuals had different multilocus genotypes with heterozygosity ranging from 0.089 to 0.843. Nine of 10 loci kept the Hardy-Weinberg equilibrium in adjusted significance thresholds. We report the development of microsatellite markers for S. exiqua potentially suitable for further studies of population structure, dispersal, and host relationship.

Microsatellite loci are increasingly used as markers in the human, animal and plant genomes. Being highly mutable, microsatellite regions are able to differentiate between related taxa, even at the level of individual isolates in a single species. Studies on mushroom population structure, gene flow and dispersal between natural and cultivated species have become central in breeding programmes and the knowledge of new polymorphic, codominant markers will be a promising avenue to exploit wild genetic resources. The molecular phylogeny in 50 different commercial cultivated strains of Pleurotus eryngii using PCR amplification with URP primers and mitochondrial microsatellite primer was studed. The sizes of the polymorphic fragments obtained were in the range of 200 to 2000 bp. RAPD analysis techniques were able to detect genetic variation among the tested strains. With these isolated PCR amplification with URP primers we intend to analyse the population structure of the P. eryngii species complex and investigate the structure of the basidiomycete genome which deserves. A few single-locus microsatellite markers have been isolated in Pleurotus eryngii and Pleurotus ferulae. This technique is useful in those species where microsatellite loci are rare in the mitochondria.

한우 이력추적제에 적용되는 11개의 MS marker (TGLA227, BM2113, TGLA53, ETH10, SPS115, TGLA126, TGLA122, ETH3, ETH225, BM1824 and INRA23)와 성감별을 위한 2개의 sexing primer로 조합된 하나의 Multiplex PCR set를 이용하여 모근에서 추출한 genomic DNA를 이용해 3510두의 대량 시료를 분석한 결과 3.93%의 genotyping 실패율로 성공적인 분석결과를 얻었다. 무작위교배집단으로 가 정 시 동일개체출현확률 (PI)은 1.31×10-23, 반형매교배집단으로 가정 시 동일개체출현확률 (PIhalf-sibs)은 2.52×10-16 그리고 전형매교배집단으로 가정 시 동일개체출현확률 (PIsibs)은 1.09×10-6으로 나타나 현재 사용 중인 11종의 MS marker는 범용적으로 사용하여도 무방할 것으로 재확인 되었다. 또한 생산 및 사 육단계의 생우의 경우 모근을 이용하여 DNA를 추출하는 것은 시료 채취 시 소에게 주어지는 스트레스를 최소화 시킬 수 있을 뿐만 아니라 대립유전자형 분석에 있어서 시간적, 경제적인 효율성을 높일 수 있었 다. 또한 모근 채취 부위 중 등, 배, 꼬리상부와 꼬리하부를 이용하여 검정한 결과 꼬리하부의 모근을 이 용하여 5~13가닥을 사용했을 때 최적의 분석결과를 보였다. 최종적으로 한우의 사육단계 대량 유전자형 분석에 적용 가능한 96 well 단위를 기본으로 하는 모근 DNA분리 체계를 확립하였다.

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.

The Laodelphax striatellus(SBPH) (Homoptera:Delphacidae) is a major rice insect pest in Asia. It brings the indirect damage by transmitting Rice Stripe Virus (RSV) and Rice Black-Streaked Dwarf Virus (RBSDV). Molecular markers for genetics studies will be very helpful for dynamics of SBPH and viruses. For surveying genetic variation among populations of SBPH by microsatellite analyses, we used an enrichment strategy to develop microsatellite markers from SBPH. Eight polymorphic microsatellite loci were selected to characterize genetic structuring of SBPH populations.

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.

The Laodelphax striatellus(SBPH: Small Brown Plant Hopper) is known to be not overwintering in South Korea and migrate from China to Korea every year by now, but these days many evidence support the possibility of SBPH overwintering. These insects do considerable damage to the rice production in Korea. If they will come from China, identification of origin of migration is very important to make countermeasure against SBHP outbreak. So we want to know exact the route of long-distance migration of them. In this study, we have developed some microsatellite markers for basic population genetic analysis of SBPH. We will show the genetic characteristics of them

The silkworm (Bombyx mori), as an industrial insect, possesses a high economic value. Casual discrimination and accumulated genetic information of silkworm varieties are essential ground for the practical utilization and long-term conservation. In this study, nine available microsatellite loci were successfully genotyped from ~50 silkworm strains preserved in Korea. According to genotyping analysis, we obtained 3 ~ 16 alleles per locus, with an average of 7.4, the observed heterozygosity ranging from 0.04 to 0.98, and the polymorphic information content (PIC) ranging from 0.06 to 0.88, revealing that some loci are highly variable. Among 54 strains 13 strains were casually identified by the presence of 17 strain-specific apomorphic alleles. Furthermore, 30 among remaining strains contained strain-specific allele combinations that are also apomorphic to each strain, allowing us to discriminate each of these from other strains by genotyping of multiple loci. These results collectively suggest that the silkworm microsatellite DNA is actually and potentially important molecular marker for the discrimination of the silkworm strains that are preserved as hundreds in Korea, as more loci are genotyped.

Ten novel microsatellite loci were isolated and characterized from the soybean aphid, Aphis glycines. The soybean aphid was recently introduced into North America where it has become a serious pest of soybeans. This invasive pest has rapidly spread throughout the midwestern United States and southern Canada since 2000. The isolated loci were polymorphic, with two to 18 alleles in 20 individuals from a single population in Korea. The analyses revealed that 19 individuals had different multilocus genotypes, showing expected heterozygosity values ranging from 0.180 to 0.891. We report the development of microsatellite markers for A. glycines potentially suitable for further studies of population structure, dispersal, and host alternation.