한우의 mt DNA cytochrome oxidase subunit I, II, 및 III complex지역의 유전적 다형현상을 제한효소를 이용하여 검출하였다. PCR primer 6종에 대하여 20가지 제한효소를 처리하였으며, Pst I, Pvu II, Rsa I, Eco RI, Bgl II, and Msp I 제한효소를 사용하여 유전적 변이를 검출하였다. 검출된 변이체와 한우의 성장과의 관련성을 조사한 결과 cytochrome oxidase subu

DNA 다형성을 이용한 누에 유전자 해석기술을 개발하기 위하여 광식성 누에 계통 J111과 비광식성계통 의 DNA를 분리하여 유전자 은행을 제작하였다. 누에 유전자 은행은 genomic DNA를 EcoRI로 절단한후 pUC18에 ligation 시켜 DH5 E. coli에 형질전환 시켰다. 형질전환 후 얻어진 colony는 15개 누에 품종의 genomic DNA에 hybridization하였을 때 누에의 품종에 관계없이 highly repetitive, moderately repetitive 및 single 혹은 low copy number 로 구분되었다. RFLP마커에 적합한 single 및 low-copy number band만을 형성하는 colony probe을 신속하게 선발하고자 colony또는 genomic DNA로 hybridization하였다. Single 및 low-copy number의 특성을 가진 219개의 clone을 선발하여 Hind III등 8종의 제한효소별로 처리한 genomic DNA를 이용하여 다형성을 검정하여 J111과 계통간 다형성을 보인 46개의 clone을 선발하였다. 선발된 clone의 일부를 J111과 를 교배하여 얻은 의 blot에 hybridization 결과 RFLP clone들이 양친검정에 이용가능하여 누에 RFLP 연관 지도 작성의 기반을 조성하게 되었다.

복숭아혹진딧물(Myzus persicae Sulzer)은 두가지 서로 다른 기주선호성을 가지는데 이 기주선호성과 형태적 특징에 기초하여 담배진닷물(Myzus nicotinae Blackman)과 담배 이외의 다른 채소류에 서식하는 복숭아 혹진딧물(M. persicae)로 분류하였지만(Blachmean, 1987) 이 분류 방법에 동의하지 않는 학자들도 많다. 이런 이유로 RAPD-PCR 기법을 이용하여 한국에 서식하는 복숭아혹진딧물에 대하여 그들의 2차 숙주선호성에 따른 DNA의 변이 정도를 살펴보았다. 실험곤충으로는 담배와 배추에서 채집하여 사육한 진딧물 각 4 clones 씩을 사용하였다. 각 clone은 한 개체를 사육하여 얻은 자손들과 그들의 후손으로 이루어졌으며, 사육한 진딧물에서 핵 DNA를 추출하고, 10개 nucleotide 길이의 random primer 100가지를 사용하여 PCR한 후 1 % agarose gel 전기영동법으로 분석하였다. 사용한 100종류의 random primer 중 83가지에서 DNA 단편이 합성되었다. 증폭된 1개의 primer당 단편의 수는 1개에서 22개였고 평균 단편 수는 약 13개였으며, 각 각 단편의 길이는 500에서 20,000 base pair사이에 분포하였다. 82가지 primer의 경우에 일부 단편의 짙기에는 차이가 있었으나 단편종류의 분포는 동일하게 나타났다. 한가지 primer경우에만 담배섭식형 1개 c clone에서 다른 7가지 clones에 없는 band가 1개 나타났다. 이때 나머지 7 clones의 단편 분포 형태는 모두 동일하였다. 따라서 이 band는 숙주 선호성과는 무관한 것으로 보인다. 결국 이 실험에 사용한 100종류의 primer에 기초하여 RAPD-PCR기법으로 DNA를 증폭한 결과 복숭아혹진딧물의 숙주선호성이 개체군간의 유전적인 차이점에 기인한다는 가설을 뒷받침할 만한 증거를 찾지 못하였다.

Deosophila melanogadter 자연집단재 alcohol dehydrogenase(ADH) allele 의 polymorphism 및 두 ADH allele 유전자형간의 적응도와 ethanol 의 상관 관계를 조사하였다. D. meanogaster의 자연집단내 ADH는 polymorphic 하였으며, FF,FS그리고 SS형의 유전자 빈도는 47.66,42.18 및 10.16%로 나타나 F 유전자의빈도가 S 유전자에 비하여 높게 분포하였다. 산란력과 우화율에서는 FF 유전자형이 SS 유전자형에 비하여 모두 약간 높게 나타났다. 자연집단에서 유래된 인공 소집단에서는 세대의 흐름에 따라 {{{{ { Adh}^{F } }}}} 유 전자형의 빈도증가와 상대적 {{{{ { Adh}^{S }}}}} 유전자형의 감소를 보였고, etha-nol은 ADH locu 상의 selective factor로서 작용함을 시사하여 주었다.

Anopheles quadrimaculatus( Say) 자매종 간의 미토콘드리아 DNA의 제한효소 절단부위변이를 Aedes albopictus의 마토콘드리아 cDNA를 probe로 이용하여 조사하였다. DNA hybridization에 의해 두 속간에는 mtDNA 영기서열의 상당한 상동성이 있음을 알 수 있었다. 개체 모기로부터 분리한 DNA를 제한효소를 사용하여 절단한 결과 자매종간에 다른 양상을 볼 수 있었으며 Hind III에 의한 mtDNA 절편만으로도 자매종들을 동정할 수 있었다.

한국산 불개미의 다형현상에 있어서 일개미의 몸의 크기와 임무수행과의 관계, 유시웅의의 시맥상의 변이, 성비 및 소의 크기에 따르는 일개미번데기의 형별발생과 일개미집단의 형별구성등에 관하여 조사한 결과를 요약하면 다음과 같다. 1. 일개미는 3형으로 분화되어 있으며, 소형은 주로 소의 내부에 동료집단의 보호와 외부에서 진딧물에 방문하는 임무를, 그리고 대형과 중간형은 주로 외부에서 식이운반, 조소 및 동료집단의 보호임무를 수행한다. 그러나 소형이 진딧물에 방문하는 임무는 반드시 고정되어 있는 것이 아니고 계절에 따라 다소 변동이 있는 것 같다.

Background : The several studies on the characteristics of Korean ginseng cultivars and breeding lines have already been carried out the level of molecular classification analysis in Korea. In spite of where Geumsan is a representative place of Korean ginseng, Geumsan native species (breeding lines) have not yet been carry out analysis of morphological, genetic characteristics and relationship. We have plan to carry out morphological, genetic characteristics and relationship for Geumsan native species, breeding lines. Furthermore, We could be used diverse genetic resources for Ginseng breeding. Methods and Results : In this study, a total of 71 breeding lines and variety (GS97-1 - Geumwon) consisting of native ginseng collections from Geumsan was analyzed to identify for Korean ginseng variety respectively, and clustered for the selection of Geumsan native ginseng in Korea using DNA markers. We collected 71 Ginseng breeding lines from Geumsan. Analyses of the genetic characteristics of the collection were conducted for extraction gDNA using sprout. We were measured DNA concentration using QIAxpert (QIAGEN). Each DNA sample was quantified at the final DNA concentration of 5 ng/㎖ using sterilized distilled water. Korean ginseng 14 variety and 57 Ginseng breeding lines from Geumsan could be identified polymorphism using the selected 6 primer (MFGp183, MFGp130, MFGp110_E, UFGp163, MFGp108 and UFGp156). Conclusion : These finding could be used for morphological and genetic characteristics for produced native ginseng in Geumsan area. Futhermore, we could be used diverse genetic resources for Ginseng breeding.

Leaf mold disease in tomato (Solanum lycopersicum) is caused by Cladosporium fulvum, a fungal leaf pathogen. One of effective ways to control leaf mold is to breed disease-resistant tomato cultivars. Cf-4 and Cf-9 resistance (R) genes encode proteins that carry a leucine rich repeat domain and are located in plasma membrane. They trigger hypersensitive response following recognition of corresponding Avr4 and Avr9 proteins of C. fulvum, respectively. Cf-4 and Cf-9 genes are originated from wild tomato species S. habrochaites and S. pimpinellifolium and have been introgressed into commercial tomato cultivars. These two highly homologous orthologs exist as a cluster with four highly homologous paralogs. Due to this reason, development of genetic markers to distinguish these two functional R genes from their orthologs and paralogs is difficult. In this study, we tried to develop single-nucleotide polymorphism (SNP) markers to select tomato cultivars carrying resistant Cf-9 genotype. The genomic sequences of resistant Cf-4 and Cf-9 alleles, susceptible cf-9 alleles, and their paralogs were obtained from the GenBank database, and two functional SNPs causing non-synonymous substitution were found among them. Based on two SNPs, the Cf-9_2-SNP-F/R primer set for high resolution melting (HRM) analysis was developed. HRM analysis with this primer set could successfully distinguish tomato cultivars carrying resistant Cf-9 allele among 30 commercial tomato cultivars, which were characterized with the gene-based marker. These indicate that the SNP marker developed in this study is useful to trace Cf-9 genotype efficiently in marker-assisted selection in tomato.

Blackleg disease caused by Leptosphaeria maculans, is the most devastating disease of Brassica germplam worldwide that causes million tonnes of crop losses per year throughout the world. To date, a total of 12 race-specific resistance genes of Brassica napus to L. maculans have been reported but linkage mapping analysis reveals that all of those loci are located in A genome i.e., in B. rapa chromosomes. B. oleracea has high ancestral synteny with B. rapa through their evolution. We believe that presence of qualitative resistance is possible in B. oleracea germplasm. The present study was therefore planned to find out any race-specific qualitative resistance gene present in C genome of B. oleracea. A total of 16 microsatellite markers were used which are linked to seven different Rlm and Lep genes of B. napus to screen 32 inbred lines of cabbage. Primers were designed based on homology assessment in corresponding nucleotide sequence available in Bolbase (a B. oleracea genome database, http://www.ocri-genomics.org/bolbase/index.html), located in B. oleracea scaffolds/chromosomes. Out of 16 SSR markers, 13 were found polymorphic which indicates possible existence of resistant genes in cabbage lines. The inbred lines are then assessed against two L. maculans stains with known avirulent genes. Some inbred lines were hypersensitive against gene-specific virulent strains of L. maculans that confirmed existence of Rlm1, Rlm2, Rlm4, LepR3 and LepR4 in the cabbage lines. In this way we were able to select out resistant and susceptible lines against each resistant gene. The gene-specific polymorphic SSR marker regions were cloned and sequenced and candidate SNPs were identified for confirmation of their functionality.

To successful molecular breeding, identification and functional characterization of breeding related genes and development of molecular breeding techniques using DNA markers are essential. Although the development of a useful marker is difficult in the aspect of time, cost and effort, many markers are being developed to be used in molecular breeding and developed markers have been used in many fields. Single nucleotide polymorphisms (SNPs) markers were widely used for genomic research and breeding, but has hardly been validated for screening functional genes in olive flounder. We identified single nucleotide polymorphisms (SNPs) from expressed sequence tag (EST) database in olive flounder; out of a total 4,327 ESTs, 693 contigs and 514 SNPs were detected in total EST, and these substitutions include 297 transitions and 217 transversions. As a result, 144 SNP markers were developed on the basis of 514 SNP to selection of useful gene region, and then applied to each of eight wild and culture olive flounder (total 16 samples). In our experimental result, only 32 markers had detected polymorphism in sample, also identified 21 transitions and 11 transversions, whereas indel was not detected in polymorphic SNPs. Heterozygosity of wild and cultured olive flounder using the 32 SNP markers is 0.34 and 0.29, respectively. In conclusion, we identified SNP and polymorphism in olive flounder using newly designed marker, it supports that developed markers are suitable for SNP detection and diversity analysis in olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

Molecular markers are useful for selecting to include superior character genetic like as strong immune system and rapid growth in fish. The marker is also very important part of breeding technology in Olive flounder (Paralichthys olivaceus). Single nucleotide polymorphisms (SNPs) marker is already in use widely for genomic research and breeding. But this SNPs marker hardly has been validated for screening functional genes in Olive flounder. We study identify single nucleotide polymorphisms (SNPs) on Expressed sequence tag (EST) database, develop usable SNP marker and apply to wild sample and cultured of olive flounder. As a result, Out of total 4.327 ESTs, 693contigs and 514 SNP from total contigs were detected while these substitutions include 297 transitions and 217 transversions. 144 developed markers were applied in 16 samples (wild 8, culture 8), Out of total marker, only 32 markers had detected polymorphic in sample. Polymorphism of 32 markers was observed in the variety genes region involved in immunity and protein synthesis. And the 32 marker were identified 21 transitions, 11 transversions, and indel was not detected in polymorphic SNPs. The analysis on heterozygosity by sample showed 0.34 in wild sample and 0.29 in cultured sample. In conclusion, we was identified SNP and Polymorphism by designed new marker, it supports that development marker is suitable for SNP detection and diversity analysis in Olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

With the development of next generation sequencing (NGS) technology, the variation of sequences represented as SNP between cultivars becomes available at genome level. The major domestic cultivars with high yield have been developed by breeding of indica and japonica, it is important to localize the region of origin according to the genotype for further characterization of unique features of cultivars. For the localization of SNP at genome level, the paired end sequences of 6 major domestic rice cultivars, Ilmi, Ilpoom, Sulgaeng, Bakjinju, Hwayoung and Woonkwang were compared against Japonica and Indica Rice Genomes as reference genomes. The genomic DNAs were prepared from callus tissues and paired-end of the fragments were sequenced with NGS Sequencer, Illumina HISeq. About 50x coverage of paired-end sequences were trimmed according to the quality of the sequences, and errors were corrected with statistical analysis of kmers of 15. The trim-corrected sequences were mapped and variants were analyzed against reference genomes. The overall change rate of Ilmi against Nipponbare IRGSP 1.0 and Indica BGI 93-11 reference genomes were 0.92 base/1kb (1/1,079 base) and 8.09 base/1kb (1 base/123 bases), respectively. Among 6 cultivars, overall rate of Bakjinju showed the lowest overall change rate of 0,53 base/1kb, and Hwayoung showed highest frequency of 0.92 base/1kb. Compared to high level in the range of change rate of 7.0-9.3 base/1kb against indica, domestic cultivars showed lower range of change rate 0.2-3.3 base/1kb with unique local high peak against japonica genome depend on the chromosomes. Compared to assembly of genome sequences, the variation of nucleotides compared to reference sequences is much faster and simple to characterize the genotype. The types of variation and the effect on functional categories will be presented.

With the development of next generation sequencing (NGS) technology, the variation of sequences represented as SNP between cultivars becomes available at genome level. The major domestic cultivars with high yield have been developed by breeding of indica and japonica, it is important to localize the region of origin according to the genotype for further characterization of unique features of cultivars. For the localization of SNP at genome level, the paired end sequences of 6 major domestic rice cultivars, Ilmi, Ilpoom, Sulgaeng, Baekjinju1ho, Hwayoung and Woongwang were compared against Japonica and Indica Rice Genomes as reference genomes. The genomic DNAs were prepared from callus tissues and paired-end of the fragments were sequenced with NGS Sequencer, Illumina HISeq2000. About 50x coverage of paired-end sequences were trimmed according to the quality of the sequences, and errors were corrected with statistical analysis of kmers of 15. The trim-corrected sequences were mapped and variants were analyzed against reference genomes. The overall change rate of Ilmi against Nipponbare IRGSP 1.0 and Indica BGI 93-11 reference genomes were 0.92 base/1kb (1/1,079 base) and 8.09 base/1kb (1 base/123 bases), respectively. Among 6 cultivars, overall rate of Baekjinju1ho showed the lowest overall change rate of 0,53 base/1kb, and Hwayoung showed highest frequency of 0.92 base/1kb. Compared to high level in the range of change rate of 7.0-9.3 base/1kb against indica, domestic cultivars showed lower range of change rate 0.2-3.3 base/1kb with unique local high peak against japonica genome depend on the chromosomes. Compared to assembly of genome sequences, the variation of nucleotides compared to reference sequences is much faster and simple to characterize the genotype. The types of variation and the effect on functional categories will be presented.

In order to select a rice population with useful trait such as arsenic tolerance for crop improvement, we have developed 3000 M7 Targeting Induced Local Lesions IN Genomes (TILLING) lines by gamma ray (GR) irradiation treatment to a rice variety (cv. Donganbyeo). A total of 2 M7 lines exhibited the arsenic (AsV) tolerant phenotype (hereafter, named Arsenic Tolerant TILLING line 1 and 2, and designed as ATT1 and 2), in which the shoots and roots length of ATT lines were significantly longer than those of wild type (WT) during As(V) treatment. To survey the DNA polymorphism of these plants, we conducted the Whole genome resequencing with 10x coverage in ATT lines. By comparative analysis among ATT lines, we have identified the common DNA polymorphism such as 11,817 SNPs (49.83% in ATT1 and 48.35% in ATT2) and 30,618 InDels (86.72% in ATT1 and 86.23% in ATT2). Also, these mutants were showed the close relationships more than WT. To further study the changed amino acids of genes, we commonly identified the 758 genes for non-synonymous SNPs and 249 genes for changed codon InDels. These genes were mainly exhibited the enriched GO functions such as catalytic activity, nucleic acid binding and transferring phosphorus-containing groups. To determine the genes associated with arsenic-related mechanism in DNA polymorphism of ATT lines, we have retrieved the two structurally altered genes (Os11g47870 and Os03g19900) for metalloid As(V) detoxification toward induced genes in response to arsenic treatments by public microarray datasets. We suggest that As(V) tolerant phenotypes of ATT lines are certainly affected by structurally altered genes associated with phosphorus transferring and As(V) detoxification during GR treatment