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.

Genotyping-by-sequencing (GBS) is a cost-efficient method which can be useful for SNP marker discovery in a population of interest. GBS is genome reduction sequencing method using restriction enzyme. The quality of DNA is a key factor which could have an influence in downstream analysis. However, there have not been many studies which investigated the impact of DNA degradation and the quality of the data on marker discovery. In this study, GBS data of 6 Hanwoo samples (H1~6) showing differing level of DNA degradation were compared. Re-sequencing pipeline was followed to investigate the impact of DNA degradation on marker discovery. As a result, we found that the quantity and quality of SNPs were not affected in the sample H5 and H6 with moderately degraded DNA. On the other hand, marker discovery was greatly affected in samples with severe DNA degradation (H3 and H4). The findings in this study support that GBS is a robust genotyping method towards moderate DNA degradation.

Microsatellite SSR markers were developed and utilized to reveal the genetic diversity of 32 strains of Flammulina velutipes collected in Korea, China, and Japan. From SSR-enriched library, 490 white colonies were randomly selected and sequenced. In the 490 sequenced clones, 85 clones (17.35%) were redundant. Among the remaining 405 unique clones, 201 clones (49.6%) contained microsatellite sequences. As a result, 12 primer pairs produced reproducible polymorphic bands within diverse 4 strains and these selected markers were further characterized in 32 Flammulina velutipes strains. A total of 34 alleles were detected using the 12 markers, with an average of 3.42 alleles and the number of alleles ranged from two to seven per locus. The major allele frequency ranged from 0.42 (GB-FV-127) to 0.98 (GB-FV-166), and values for observed (HO) and expected (HE) heterozygosity ranged from 0.00 to 0.94 (mean = 0.18) and from 0.03 to 0.67 (mean = 0.32), respectively. SSR loci amplified with GB-FV-127 markers gave the highest polymorphism information content (PIC) of 0.61 and mean allele number of five, while for loci amplified with GB-FV-166 markers these values were the lowest, namely 0.03 and two. The mean PIC value (0.29) observed in the present study with average number of alleles (3.42). The genetic relationships among 32 Flammulina velutipes strains based on SSR data were generated by UPGMA cluster analysis. In conclusion, we succeeded in developing 12 polymorphic SSRs markers from SSR-enriched library of Flammulina velutipes. These SSRs are presently being used for phylogenic analysis and evaluation of genetic variations. In future, these SSR markers will be used in clarifying taxonomic relationships among the Flammulina velutipes.

This study was conducted to evaluate the genetic distances and specific DNA makers by the randomly amplified polymorphic DNA (RAPD)-PCR method for the Korean native chickens(red plumage, red-line plumage, Ogol) and white leghorn. Genomic DNA was extracted from plumage from chickens after they were slaughtered. The extracted DNA was observed by nano-spectrometer. RAPD analysis was performed using 13 different primers. Statistical analysis was made for the estimation of the genetic distance among the chicken’s and the cluster tree was drawn by using MEGA 5.05 software. Genetic relations among them were determined by RAPD analysis. The polymorphic bands were observed 72% and the rest of 28% was monomorpic. The largest genetic distance (2.266) was found between the native chickens(red, red-line) and the ogol chickens by UPGMAP method and the closest distance was observed between the ogol in korean chickens as expected. The highest genetic distance between them was estimated 2.266 and in the dendrogram analysis, among I and II within cluster II, most of the ogol chickens were in IIB, indicating the expression of the ogol color could be due to original and the ancestral genetic crossing. Thus, this genetic distance can be useful as the differential genomic information in the normal(red plumage, red-line plumage) and ogol of korean native chickens. This work was supported by a grant from the “Livestock Preservation of Genetic Resources", Rural Development Administration, Republic of Korea.

Next Generation Sequencing을 이용한 분석 서열을 기반으로 매미나방의 Microsatellite loci 탐색 및 marker 개발을 수행하였다. 매미나방의 Genomic DNA 서열 분석은 MiSeq Sequencer (Illumina)의 1/8 plate를 이용하여 실시하였다. 판독된 유전자 서열의 길이는 총 3,974,358,483 bp로 평균 248.58 bp로 구성된 총 15,988,036 개의 분석 단편이 확보되었으며, 이를 CLC workbench를 이용하여 총 367,397,618 bp로 조합하였다. 조합된 Genomic DNA 서열을 대상으로 반복서열길이 2~4 bp, 반복횟수 4회 이상의 조건으로 총 1,864 개의 Microsatellite loci를 탐색하였다. 이 중 반복횟수 6회 이상의 430 loci에 대한 marker 제작 가능성을 TM 55.5~56.5℃, GC contents 30% 이상, primer length 18~22 bp의 조건으로 Primer3을 이용하여 분석하였으며, 총 207 개의 marker를 제작하였다. 선별된 207개 marker 중 150개 마커에 대해 일반 올리고 primer set를 제작하여 PCR을 통한 유용성 평가를 실하였으며, 그 결과 총 29개의 마커에 대한 유효성이 확인되어 Genotyping 용 형광 dye인 FAM을 부착한 분석용 마커로 제작하였다. FAM을 부착한 마커에 대한 PCR 효율 검사를 통해 최종적으로 10개 마커를 선별하여 한국 4개 지역(Korea 1, Korea 22, Korea 26, Korea 31) 및 러시아(Vladivostok), 몽고(Shagaarnur) 각 1개 지역의 개체군을 대상으로 유전적 구조 분석을 수행하였다. 유전적 유사도를 평가하기 위하여 Fst Pairwise UPGMA tree를 분석한 결과, Korea 1과 러시아 개체군, Korea 22와 Korea 26 개체군의 유전적으로 유사도가 높은 것으로 나타났으며, Korea 31과 몽고 개체군은 유사도의 기부에 위치하는 것을 확인할 수 있었다. 또한, Baysian Algorithm을 기반으로 한 유전적 구조 분석에서도 각 개체 및 개체군의 구조는 UPGMA tree 동일한 양상을 나타내는 것으로 확인되었다. 따라서, 현 연구를 통해 개발된 매미나방의 Microsatellite 마커는 한국을 비롯한 인근 지역의 지역적 개체군 분석을 가능하게 할 수 있을 것으로 판단되며, 결국 식물검역에서 매미나방의 유출 국가 및 지역에 대한 판별 분석에 유용할 수 있을 것이다.

In the present study, the genetic diversity of 69 Ganoderma species from various regions was determined by different molecular markers, including the internal transcribed spacer (ITS) rRNA, a partial β-tubulin gene, and mitochondrial small-subunit ribosomal (SSU) rDNA gene as well as randomly amplified polymorphic DNA (RAPD) analysis. The size of the ITS rDNA regions and mitochondrial SSU rDNA gene from different Ganoderma species varied from 625 to 673 bp and 656 to 2,040bp, respectively, and those of the partial β-tubulin gene sequence were 419 bp. Phylogenetic analysis based on the ITS region, the partial β-tubulin gene, and the mitochondrial SSU rDNA gene reveal that Ganoderma species are largely divided into two groups. Interestingly, most of the Ganoderma lucidum strains could be classified into 1 group, while other Ganoderma species divided into several groups (4 or 5 groups) in phylogenetic tree. One fragment unique to G. lucidum was selected from the RAPD profile and then sequenced. One primer pair (designated as KGS-F and KGS-R) based on this specific fragment was designed to amplify a 559 bp DNA fragment within the sequenced region. A single band with the expected size of 559 bp was observed from G. lucidum, except for G. lucidum strains from China, Canada, and Taiwan. This specific marker for G. lucidum from RAPD analysis, also supported by the phylogenetic analysis of the ITS, partial β-tubulin gene, and mitochondrial SSU rDNA sequences, will be useful for the PCR-based identification of G. lucidum in research applications as well as in the market.

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 soybean Kunitz trypsin inhibitor (KTI) protein is responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. Ti locus controls presence or absence of Kunitz trypsin inhibitor protein. Genetic recombination or tight linkage between Ti locus and Satt228 marker that has been identified to be tightly linked to the Ti locus was detected for marker assisted selection (MAS) using two F2 populations of titi genotype in this study. Two F2 populations were developed from the cross of A29 (KTI protein present, TiTi genotype, AA genotype in Satt228 marker) x Gaechuck#1 and Gaechuck#2 (KTI protein absent, titi genotype, BB genotype in Satt228 marker). Among 31 F2 plants derived from A29 x Gaechuck#1, twenty nine F2 plants show BB genotype that indicates no recombination between Satt228 marker and Ti locus. Only 2 F2 plants show AA genotype that indicates recombination between Satt228 marker and Ti locus. Twenty eight F2 plants derived from A29 x Gaechuck#2 show BB genotype that indicates no recombination between Satt228 marker and Ti locus. Expected genetic ratio between Satt228 marker and Ti locus was 3.6 cM in F2 population.

한국꿩 (Korean ring-necked Pheasant, Phasianus colchicus karpowi)과 외국 아종의 유전적 유연관계를 파악하기 위해 야생 한국꿩, 사육 한국꿩, 사육 한국꿩과 외국꿩간의 잡종꿩, 외국꿩 4아종(중국 링넥, 흑 뮤탄트, 백 뮤탄트, 녹치)을 대상으로 ISSR 표지자 분석과 AMOVA 분석을 수행하였다. 야생 한국꿩의 전체 유전 다양성중 94.08%가 서식지 내 개체간 유전적 차이에 기인하고, 5.9% (Φ

Hedera helix 11계통, Hedera rhombea 3계통, Fatshedera lizei 1계통, 그리고 Fatsia japonica 1계 통을 수집하였다. RAPD primer 10개를 이용하여 수 집된 3속의 재료들의 유전적 다양성을 측정하였다. 3 속을 재료로 사용하여 밴드간 96.9%의 높은 다형성을 보였다. 총 97개의 RAPD 밴드를 이진화하여 UPGMA 방법을 이용하여 계통도를 작성하였다. Hedera helix 계통들은 모두 1개의 그룹에 속하였으며 8계통의 유전 적 거리는 극도로 적었으며 나머지 3계통 역시 유전적 으로 가까웠다. 하지만 형태적으로는 높은 다형성을 보 였다. 따라서 수집된 유전자원을 이용한 돌연변이체 개 발이 가능성이 있는 방법으로 제시되었다. Fatsia japonica는 유전적으로 관계성이 적어 다른 종들과 평 균 0.63의 유전적 거리를 보였다. Hedera helix와 Fatsia japonica 속간 교배를 통하여 개발된 Fatshedera lizei는 Hedera rhombea 계통들과 함께 계통도에서 위치하였다.

과피와 과육의 다양한 색은 복숭아 분류에 가장 널리 사용되는 상업적 기준 중 하나이다. 새로운 적색 과육 품종을 육성하기 위해서는 많은 교배 조합과 세대가 진전되어야 한다. 따라서 육종 효율을 높이기 위해서는 목적 형질을 가진 개체에 적용할 조기 선발 분자표지를 개발할 필요가 있다. 과육색이 다르게 발현되는 복숭아 품종의 유전자 발현을 비교하기 위해 2개의 cDNA library를 제작하였다. 적색 과육 품종인 ‘조생혈도’와 백색 과육 품종인 ‘미백도’의 유전자 발현 차이를 보기 위해 차세대 염기서열 분석(NGS) 기술을 사용하였고, 두 품종으로부터 얻은 EST의 염기서열을 결정하고 기존에 보고된 유전자와의 상동성을 분석하였다. ‘조생혈도’와 ‘미백도’의 EST database로부터 72쌍의 SNP 분자표지를 선발하였고, 적색 과육 품종 8개와 백색 과육 품종 24개를 구분할 수 있는 SNP 분자표지를 HRM 방법으로 분석하였다. 본 연구에서는 복숭아 EST database를 기반으로 HRM 분석 방법을 이용하여 복숭아 품종의 적육계와 백육계를 구분할 수 있는 효율적인 SNP 분자표지를 개발하였다. 이러한 SNP 분자표지는 복숭아 육종에 유용하게 사용할 수 있으며, 복숭아 품종의 다양한 색 변화에 관한 분자 기작 연구에 좋은 참고자료가 될 수 있을 것이다.

Bulb onion (Allium cepa), which belongs to the family Amaryllidaceae, is one of the oldest vegetative crops known to humans. Despite its high economic value, only a few reports are available on the use of molecular markers in genetic diversity analysis of Allium cepa for its improvement. Molecular genetic markers have been widely used as powerful tools for analyzing the plant genome. In particular, Microsatellites or simple sequence repeats (SSRs) markers are tandem repeats of one to six bp in length and have been proven to be the most powerful polymerase chain reaction (PCR)-based DNA markers in plant diversity analysis. In this study, the genomic DNA was isolated from different Allium cepa lines. The ESTs and gDNA sequences of onion were collected from National Center for Biotechnology information. The SSRs with two to five motifs over a length of 12 bp, were identified using SSRIT (Gramene) software. The PCR products of 100 to 350 bp in length containing SSRs, primers was designed using Primer3 with lengths of 20 to 24 bp and a melting temperature of 60℃. The SSR markers with high polymorphism-information content (PIC) levels was useful for collecting progeny with high genetic homogeneity for onion breeding, and to obtain representative marker sets for genetic tests. The SSR Finder program and the developed SSR markers could be a useful resource for genetic diversity and purity testing in onion.

Maize is one of the most important food and feed crops in the world including Southeast Asia. In spite of numberous efforts with conventional breeding, the maize productions remain low and the loss of yields by drought and downy mildew are still severe in Asia. Genetic improvement of maize has been performed with molecular marker and genetic engineering. Because maize is one of the most widely studied crop for its own genome and has tremendous diversity and variant, maize is considered as a forefront crop in development and estimation of molecular markers for agricultural useful trait in genetics and breeding. Using QTL (Quantitative Trait Loci) and MAS (Marker Assisted Breeding), molecular breeders are able to accelerate the development of drought tolerance or downy mildew resistance maize genotype. The present paper overviews QTL/MAS approaches towards improvement of maize production against drought and downy mildew. We also discuss here the trends and importance of molecular marker and mapping population in maize breeding.

무핵 포도 품종 육성 시, 주요 교배친으로 사용되는 무핵 품종 20점의 유연관계를 분석 하고, 무핵 형질과 연관된 P3_VvAGL11 마커의 이용 가능성을 검토하였다. SSR 마커 30종을 이용한 결과 218개의 대립인자가 확인되었고, 마커 당 대립인자 수는 평균 7.3개였다. PIC (Polymorphism information contents) 값은 SSR 마커에 따라 0.052에서 0.883으로 나타났고 평균 값은 0.442이었다. 또한 P3_VvAGL11 마커를 이용하여 포도 무핵 유전자원 20 점을 분석한 결과 ‘Sultanina’와 ‘Kishmish Chernyi’ 품종으로부터 유래한 18개 무핵 품종에서 무핵 특성 특이적인 밴드가 증폭되었다. 선발된 218개의 다형성 밴드를 이용하여 UPGMA 군집분석한 결과 유사도 지수 0.722를 기준으로 4개의 그룹으로 구분할 수 있었다. 각 그룹은 교배친으로 사용된 품종들의 유전적 background에 유럽종(V. vinifera), 미국종(V. labrusca), 그리고 이들의 종간교잡종인 V. labruscana의 비율에 따른 분류와 일치하였다. 품종 간 유전적 유사도는 0.592에서 0.844의 범위로 나타났으며 평균 유사도 지수는 0.715였다. 가장 높은 유사도 지수를 나타낸 것은 ‘Emerald Seedless’와 ‘Ruby Seedless’ 품종 간이었고, 가장 낮은 유사도 지수를 나타낸 것은 ‘Beauty Seedless’와 ‘Honey Seedless’ 품종 간이었다. 이러한 결과는 무핵 포도 품종 육성 시, 유전적 background가 다양한 양친 선정 및 무핵 교배 실생 조기 선발에 활용 될 수 있을 것이다.

Rice genetic resources are composed of various species and ecotypes, and each accession reveals different genetic and phenotypic characters. For the managenment of diverse rice genetic resources, seed integrity is important factor in that the individuals of one accession in self-pollinating crop might be homogeneous. To elevate the management efficiency of rice germplasm contrary to the phenotypic distinction, we focused on applicable microsatellite markers because this markers are widly used for genetic evaluation in diverse genetic resources with a character of high reproducibility and polymorphism. In this regard, we selected microsatellite markers based on genotypes; diversity set including 150 accessions using 249 SSR markers. As SSR loci with high PIC(polymorphism information content) values usually revealed multi bands in one accession, proper genotyping were difficult in these loci. Therefor, we checked the band clarity in addition to PIC values and chose 12 and 6 SSR markers finally. All accessions of rice diversity set were distinguished with the first marker set comprising 12 SSR markers, and only 3 combinations of tested accessions(0.03%, 3/11,175) showed same genotype with second marker set comprising 6 SSR markers. The tested 142 Korean bred varieties revealed 0.19%(19/10,011 combinations) and 0.69%(69/10,011 combinations) genotypic identity using first and second marker set, respectively. These newly selected markers might be useful for the analysis of genetic homogeneity and relationship in rice genetic resources.

Molecular genetic markers have been widely used as powerful tools for analyzing the genome. In particular, SSR markers have practical applications in breeding systems because they can be used in high-throughput analyses for genetic mapping, heritable diversity testing, purity analysis, and marker-assisted selection. Currently, due to technical advances in DNA sequencing, large sequence databases are available for large-scale SSR mining and marker development. Here, we describe an automated method, the SSR Finder program, for SSR discovery in the onion sequence database, and primer design for amplifying the detected SSRs. A total of 1,049 SSR primers were obtained for genetic purity testing, and 100 SSR sets were analyzed in 14 bulb onion breeding lines using clustering analysis. A total of 20 selected markers from screening of all 1,049 SSR primers, were finally applied for genetic purity testing in three breeding lines, NW1, NW9, and NW10. The initial tests showed that 15%, 71%, and 97% of individuals within NW1, NW9, and NW10, respectively, were genetically homogeneous. These markers produced using the SSR Finder will be useful for investigating the genetic purity of onion breeding lines.

In this study, Expressed Sequence Tag-Simple Sequence Repeat (EST-SSR) analyses were used to clarify the genetic polymorphisms among Korean ginseng cultivars and breeding lines and to classify them into distinct genetic groups. Polymorphic and reproducible bands were produced by 14 primers out of total 30 primers used in this study. Fourteen EST-SSR loci generated a total of 123 bands. Amplified PCR products showed the highly reproducible banding patterns at 110~920 bp. The number of amplified bands for each EST-SSR primers ranged from 2 to 19 with a mean of 8.8 bands. P26 and P35 primers showed 13 and 12 banding patterns, respectively. The number of alleles for each EST-SSR locus ranged from 1.67 to 2.00 with a mean of 1.878 alleles. P34 and P60 primers showed the highest and the lowest genetic polymorphism, respectively. Cluster analysis based on genetic similarity estimated by EST-SSR markers classified Korean cultivars and breeding lines into 4 groups. Group included Gopoong and Chunpoong and 9 breeding lines (55%), group included 2 breeding lines (10%), group included 3 breeding lines (15%), group included Gumpoong and 3 breeding lines (20%). Consequently, the EST-SSR marker developed in this study may prove useful for the evaluation of genetic diversity and differentiation of Korean ginseng cultivars and breeding lines.