The demand for novel strains has been rising in the domestic market to increase the production of sclerotia from Wolfiporia hoelen. To improve strain breeding efficiency, we investigated whether single-nucleotide polymorphisms (SNPs) in the RNA polymerase II subunit (RPB2) gene, which may be linked to the mating type locus, are useful for distinguishing monokaryons from dikaryons in Korean W. hoelen strains. We designed a specific primer set to efficiently amplify a region of RPB2 using PCR with the genomic DNA of 12 cultivated strains and 31 wild strains of W. hoelen collected from Korea. Nucleotide sequences of the PCR-amplified RPB2 genes were determined and analyzed for the presence of SNPs among the 43 W. hoelen strains. Previously reported SNP loci were detected in the RPB2 gene of all W. hoelen strains tested. However, these previously reported SNP loci could not be applied to differentiate monokaryons from dikaryons in approximately one-third of Korean wild strains with homozygous genotypes. Three additional SNPs in the RPB2 gene, which may improve the ability to distinguish monokaryons from dikaryons, were identified by searching through the multiple sequence alignments of the 43 W. hoelen strains. The applicability of these three novel SNPs, together with the previously known SNPs, in the RPB2 gene to W. hoelen strain breeding was verified by examining the hybrid strains and their parental strains.

미토콘드리아 시토크롬 c 산화효소 1 (COX1) 유전자 염기서열(658 bp)을 사용하여, 콩 포장에서 채집된 어리팥나방(Matsumuraeses falcana)과 팥나방(Matsumuraeses phaseoli)의 종을 실험실 집단의 종들과 비교하여 동정하였다. COX1 염기서열 분석에서, 어리팥나방 47개체 로부터 10개의 하플로타입이 발견되었고, 종내 유전적 거리는 0.15~0.46%이었다. 이중 하프로타입 A형이 약 70%로 우점형이었다. 팥나방의 30개체로부터는 모두 동일한 하나의 서열만이 확인되었고, 어리팥나방과의 종간 유전적 거리는 4.11~4.61%이었다. 두 종의 COX1 염기서열을 번역한 아미노산 서열은 모두 동일하여 동의적 염기서열 변이(동의치환, 同義置換, synonymous substitution)를 확인할 수 있었다. 포장 조사에 서 두 종의 유충이 콩의 잎과 꼬투리를 가해하였고, 한 포장에서 동시에 발생하였다. 전체 포장에서 어리팥나방의 평균 밀도는 팥나방보다 약 1.5 배 높았다. 이 결과는 콩이 두 종의 동일 기주임을 명백하게 제시하였다. 별도로 이 속의 유충 기생파리로서 Elodia flavipalpis (파리목: 기생파리 과)가 발견되었고, COX1 서열로 동정되었다.

밀은 세계 3대 주요 작물이지만 우리나라는 대부분 수입에 의존하고 있으며, 자급률이 1%도 도달하지 못한 실정이다. 국내 밀 품종은 40여 종 이상으로 지속적으로 개발되고 있으며, 품종마다 용도와 농업적 특성이 달라 목표 형질에 맞는 적절한 품종을 선택하여 재배하는 것이 중요하다. 품종을 정확하고 빠르게 판별하기 위해선 분자 마커 기술이 필요하다. 분자 마커는 생육 환경과 시기에 영향을 받지 않고 다양한 품종을 신속하고 정확하게 구분할 수 있어 유전적 변이성, 농업 형질 등을 분석하기에 유용하다. 본 연구는 기존에 보고된 국내 밀 32품종에 대한 SCAR (Sequence Characterized Amplified Region) 판별 마커를 재검정하여 재현성이 높은 마커를 선별하였으며, 기존에 검정되지 않은 국내 밀 9품종에 추가 적용하여 비교분석하였다. 15개의 마커 세트 중 6개의 마커가 재현성과 정확도가 높은 것으로 확인되었고, 최종적으로 국내 41품종 중 4, 7, 8, 10, 11, 12번 마커를 이용하여 다홍, 금강, 밀성, 조은, 수강, 한백, 조광, 영광을 판별할 수 있었다. 또한, 4번 마커를 통한 증폭산물의 염기서열 분석을 통해 SNP (Single Nucleotide Polymorphism)를 발굴하여 ‘올밀’에 대한 새로운 품종 판별 마커인 SdHRM1과 SdHRM2를 개발하여 HRM (High Resolution Melts) 분석을 시행하여 육안으로 판단하기 어려운 SNP를 정확하게 판별할 수 있었다. 품종간의 SNP를 활용한 품종 마커 기술은 다양한 농업형질에 적용할 수 있으며, 분자 육종 프로그램에 실질적인 도움이 되는데 큰 기여를 할 것으로 사료된다.

This study identified single nucleotide polymorphisms (SNPs) that affect the body weight of chickens. Analysis of body weight showed that the Cornish breed had the highest body weight, and the Korean native chicken (Gray Brown) had the lowest body weight. TSH is composed of an α-subunit and a β-subunit, and the TSH-β gene encoding the β-subunit has been reported to be associated with obesity. In chickens, it is located on chromosome 26 and is reported to be associated with growth. The calcium-sensing receptor gene (CaSR) plays a role in the regulation of extracellular calcium homeostasis and is responsible for calcium absorption in the urinary tract, which affects the eggshell quality in poultry. It was shown that TSH-β was strongly correlated with weight in Cornish and Korean native (Gray Brown) chickens, particularly in those with the CC trait. However, CaSR showed no association with body weight in poultry; it was associated with calcium and the eggshell. Thus, selection for TSH-β can be used to produce individuals with more favorable traits in terms of body weight.

본 연구는 한우와 함께 우리의 고유유전자원임에도 불구하고 한우에 비해 산업적 활용이 미흡한 칡소집단의 유전적 다양성과 특성을 파악하고자 실시하였다. 또한 멸종위기까지 처했던 칡소가 복원사업을 통해 일정 수준의 축군 확보에 성공하였지만 이 과정에서 유전적 다양성의 증감에 따른 유전적 병목현상 등의 현황을 파악하기 위해 11개의 MS 마커 유전자형을 이용해 분석을 수행했다. 관측(HObs) 및 기대 이형접합율(HExp)의 평균은 0.753, 0.765였으며, 다형정보지수(PIC)값은 0.732로 유전적 다양성은 비교적 높게 유지되고 있었다. 하디-바인베르크 평형(HWE) 검정결과 11개 좌위 중 5개 좌위는 유전적 평형 상태에서 유의적으로(p<0.05) 이탈해 있음을 확인했다. 칡소집단의 병목현상 여부를 검정하고자 11개 좌위의 대립유전자형을 3가지 변이 모델 IAM (Infinite Allele Model), SMM (Stepwise Mutation Model) 및 TPM (Two-Phase Mutation Model) 방법으로 추정했다. 본 연구에서 세 가지 검정 모두 칡소집단이 변이부동평형(Mutation Drift Equilibrium)에서 유의적으로 높게 이탈해 있음을 보였으며 이는 최근 유전적 병목현상이 발생했음을 시사한다. 희소 대립유전자 발생비율 검정결과 칡소 집단은 유전적 병목현상이 발생했음을 시사하는 모드변화(mode shifted) 분포를 보였다. 본 연구는 칡소를 대상으로 유전적 병목현상 여부를 분석한 최초의 보고이며 제시된 자료들은 새로운 육종소재로서 칡소의 활용과 이를 기반으로 한우산업의 다양한 육종 및 브랜드화 전략을 수립하는데 기초자료로 유용하게 활용 될 것으로 기대된다.

본 연구에 이용되는 국내 재래 흑염소는 유전적 자원을 보존하고 품종을 유지하기 위하여 소규모 집단에서 고도의 근친교배를 수행하고 있어 유전적 자질의 다양성이 저해되고 있을 것으로 우려되었다. 따라서 본 연구는 국내 재래흑염소 경상대계통 집단의 유전체 정보를 활용하여 재래흑염소 집단의 유전적 특성과 유효집단의 크기를 추정하고자 실시하였다. 국내 재래 흑염소의 유전체 정보는 Illumina Goat SNP 50k chip (illumina, inc., San Diego, CA)의 정보를 분석하여 연구에 이용하였다. 각 염색체의 인접 표지인자 와의 연관불평형 (Linkage Disequilibrium)은 0.225로 추정이 되었다. 또한, 표지인자 사이의 거리가 증가함에 따라 연관불평형의 값은 감소되는 것으로 나타났다. 국내 재래 흑염소의 유효집단크기는 최근의 세대로 오는 경우 감소하는 추세를 보였으며, 13세대에서 유효집단의 크기는 29두로 추정되었다. 재래 흑염소 경상대계통은 낮은 연관불평형 값과 유효집단 크기를 보여 유전적 자원의 다양성이 낮을 것으로 보이며, 이를 해결하기 위한 계획적인 교배와 품종 집단의 크기를 키우기 위한 대책이 필요할 것으로 사료된다.

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Recently, polymorphisms reported to be significant association with sperm MOT. This study was conducted to evaluate the SNP in the coding region of ESR1 (g.672C>T inexon 1) as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results,The g.672C>T was significantly associated with frozen semen motility and kinematic characteristics. g.158 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP Also, the SNP was low significantly associated with ALH.Therefore, we suggest that theSNP in the coding region of ESR1 (g.672C>T in exon 1) may be used as a molecular marker for Duroc boar Post-thawed semen quality.

To control an external parasitic mite, a honey bee line possessing high hygienic behavior (HHB) against an external parasitic mite, Varroa destructor, has been bred in South Korea and an assessment method has been necessitated to diagnose HHB line from the low hygienic behavior (LHB) line. Thus, in this study, we developed single nucleotide polymorphism (SNP) markers from whole genome sequencing of each 20 worker bees from HHB and LHB lines of A. mellifera ligustica (Hymenoptera: Apidae). An average of 319,445,977 sequence reads was mapped to the known A. mellifera reference genome (an average of 87.46%). In 2,316,128 and 3,266,756 SNPs from each HHB and LHB line, an average of 93.6% and was located in the intergenic spacers and introns, whereas, the remaining 6.4% was located in the genic region, respectively. Among them 20 SNPs that were fixed at each line possessing within-individual homozygosity were selected and each four SNPs were used to diagnose the two honey bee lines either by typical PCR-restriction fragment length polymorphism method or allele-specific PCR. The remaining six SNPs had the size difference, enabling relatively easy differentiation between the two honey bee lines on typical agarose gel and another remaining six SNPs only has sequence difference including SNP sites. Thus, these SNP markers can be used to diagnose the honey bee line with HHB from LHB line against V. destructor.

본 연구는 PCR 기반 RFLP (Restriction Fragment Length Polymorphism; 제한절편 길이 다형성) 분자기법을 활용하여 난 및 치어 대상 납자루아과 어류 3종의 동정을 좀 더 빠르고 정확하게 파악하고 납자루아과 어류의 종별 산란양상 및 번식생태 이해에 대한 기여가 목적이다. 본 연구를 위해 기존 선행된 문헌자료를 확인하고 납자루아과 어류가 2종 이상 동서하고 있는 지역을 확인하여 현지조사를 수행하였다. 현지조사 결과 확인된 납자루아과 어류는 묵납자루 (Acheilognathus signifer), 줄납자루 (A. yamatsutae) 및 각시붕어 (Rhodeus uyekii)로 총 3종이 확인되었으며, 확인된 납자루아과 어류와 동서하고 있는 숙주조개 (작은말조개; Unio douglasiae sinuolatus)를 채집하여 숙주조개 속 납자루아과 어류의 난 및 치어를 확보하였다. 현지조사 결과 확인된 납자루아과 어류 3종을 대상으로 미토콘드리아 DNA COI과 cyt b 유전자 염기서열을 비교하여 각각 종별로 특이성을 지닌 부위 (단일염기변이; Single Nucleotide Variation: SNV)에 맞는 제한효소를 선정하였고, 숙주조개 속 난 및 치어를 대상으로 genomic DNA를 추출하여 PCR-RFLP 실험을 수행한 결과 현지조사 시 확인된 납자루아과 어류 3종의 독특한 제한절편 길이 양상을 전기영동을 통하여 확인하였다. 본 연구를 통해 묵납자루, 줄납자루 및 각시붕어의 종을 판별할 수 있는 RFLP 마커를 개발하였으며, 숙주조개 난 및 치어를 대상으로 정확한 종의 동정을 보다 빠르고 효과적으로 수행하여 각각 납자루아과 종별 산란양상을 보다 정확히 규명하고 향후 이들 자연개체군의 효과적인 유지, 관리 및 보전 방법 개발에 유용하게 활용될 수 있을 것으로 판단된다.

Hypsizygus marmoreus is a mushroom with abundant flavor and medicinal properties. However, its application is limited by problems such as long cultivation period, low biological efficiency, and microbiological contamination; therefore, there is a substantial need for development of new cultivars of this species. In this study, 55 strains of H. marmoreus were subjected to inter simple sequence repeat (ISSR) analysis to identify markers for the selection of mother strains for breeding from the collected germplasm. ISSR 13 and 15 were confirmed as polymorphic markers. The three strains (KMCC03106, KMCC03107, and KMCC03108) with white cap color were found to be genetically closely related upon UPGMA analysis of both ISSR 13 and 15. Based on the PCR analysis results for ISSR 15, the collected germplasm were differentiated into three groups according to the strain collection year. Thus, ISSR 15 could be a marker for determining the phylogeny of cap color and genetic variations according to the strain collection year. These results suggest that ISSR markers can be effective tools for the selection of mother strains for breeding of H. marmoreus.

Volvariella volvacea is mainly cultivated in subtropics area like South-East Asia. Because it is cultivated in rice straws, it is called a straw mushroom. That mushroom grows well in high temperature about 30~38°C and high humidity. Straw mushroom is a homothallic mushroom, so it is difficult to identify whether the offspring is different from the parents. This study was carried out to investigate RAPD primers that can be used for identification the DNA polymorphism of Volvariella volvacea genetic resources. 9 strains were collected from various countries like China, Vietnam etc. When ITS regions of their DNA were analyzed, they proved to be Volvariella volvacea. A cultivation test was conducted to measure the morphological characteristics of them 2 strains, KMCC04380 and KMCC04382 were selected for breeding resources because the mycelium of them grew well on medium and fruiting bodies were formed quickly. The Universal PCR Fingerprinting kits(UPF primers) were used to confirm the genetic polymorphisms of the 2 strains. As a result of confirming the DNA bands, 2 of 12 primers could be used to genetically distinguish 2 strains. About 50 spores were isolated from their fruiting bodies respectively and they also will be confirmed DNA polymorphisms by using UPF primers.

H. mamoreus is a mushroom with abundant favor and medicinal use. However, its cultivation has problems such as the long cultivation period, low biological efficiency and microbiological contamination, so new cultivars should be developed. In this study, 55 strains of H.marmoreus were analyzed with ISSR markers to identify precise genetic information in collected germplasm as mother strains in breeding. ISSR 13 and 15 markers were confirmed polymorphism. The three strains (KMC03106, KMC03107, and KMC03108) with white cap color were close genetic relationship in the UPGMA analysis of both the ISSR 13 and 15 markers. Especially in the PCR result of the ISSR 15, the collected germplasm were differentiated to three groups following collecting year. As these results, the ISSR 15 marker would be able to classify the phylogeny of cap color and genetic variation along the collecting year. Therefore ISSR markers will confer effective selection of mother strains for breeding of H. mamoreus.

Abnormal HLA-G expression occurs in various diseases such as melanoma, renal cell carcinoma, asthma, and classic Hodgkin’s lymphoma. The purpose of this study was to determine whether HLA-G gene is linked with oral squamous cell carcinoma (OSCC). To investigate the possible link with susceptibility to OSCC, 54 OSCC patients and 120 healthy controls were enrolled in this study. HLA-G 14bp insertion/deletion polymorphism is in 3'-untranslated region of HLA-G gene. HLA-G 14bp insertion/deletion polymorphism was analyzed using the polymerase chain reaction (PCR) method. For the analysis of genetic data, SPSS18.0 program was used. Logistic regression models were performed for odds ratio (OR), 95 percent confidence interval (CI), and p value. There was a significant difference in distribution allele between OSCC patients and control subjects (OR=0.018, 95% CI=0.002- 0.131, p<0.001). Our results suggest that HLA-G 14bp insertion/deletion polymorphism may be linked with susceptibility to OSCC in the Korean population.

We developed single nucleotide polymorphism (SNP) markers and are establishing diagnostic systems to distinguish disease resistance- and susceptible-strains of honey bees using the SNPs. For development of SNP markers, whole genome was sequenced each from 20 individuals of “disease resistance-strain” and “susceptible-strain” of Apis mellifera ligustica using the Illumina HiSeq 2000 sequencer. Approximately, 344 and 294 million sequence reads were mapped to the honeybee reference assembly (Amel_4.5) for each strain, respectively. Among the total 2,246,428 SNPs yielded, 33 were found to be fixed between the two strains with all homozygosity. Sixteen of them were casually amplified and sequenced from randomly selected each 10 individual of honey bees from each strain and presented strain specific SNPs. These ten SNPs were used to diagnose the two strains either by original size difference, caused by indel-accompanying SNP, typical PCR-RFLP, or AS PCR.

Oriental fruit fly, Bactrocera dorsalis, is one of the serious pests of fruit throughout the world. Two regional populations (Bangladeshi and Malaysia) were collected using methyl eugenol lure and one laboratory population was obtained from Thailand. Three populations were compared in cytochrome b (CYB) cytochrome oxidase I (COI) sequences. COI sequences were highly homologous. However, CYB sequences were polymorphic and used to discriminate these regional populations by a phylogenetic analysis. In CYB phylogeny analysis, Malaysia and Thailand populations were clustered and Bangladeshi population was separated

This study aimed to indentify single nucleotide polymorphisms (SNPs) in exon region of the glycerol-3-phosphate dehydrogenase 1 (GPD1) gene and to evaluate their associations with meat yield and quality traits in Hanwoo (Korean cattle). Two SNP markers, g.2766C>T and g.5105A>G were identified in the exons 5 and 8, respectively. Genotyping of the two SNPs was carried out using PCR-RFLP analysis in 309 Hanwoo steers to evaluate their associations with meat yield and quality traits. As a result, g.2766C>T in exon 5 was significantly associated with carcass weight (CW) and marbling score (MS). Animals with the CC genotype of g.2766C>T had heavier CW than those with the CA or AA genotype. Animals with the CC genotype of g.2766C>T also had higher MS than those with the CA or AA genotype. Additive effect was also observed with CW and MS traits. We constructed haplotypes by linkage disequilibrium analysis and analyzed association between haplotypes and meat yield and quality traits. Haplotype of GPD1 gene was associated with CW. As a result, animals with CA haplotype had heavier CW than TG haplotype. These findings suggest that the SNPs of bovine GPD1 gene may be a useful molecular marker for selection of meat yield and quality traits in Hanwoo.