초록 ▼

농업유전자원의 보존 • 관리 및 이용에 관한 법률 제16조에 의거 가축유전자원에 대한 책임기관(국립축산과학원 가축유전자원시험장)과 관리기관(11개소)이 지정·운영으로 각 관리기관에 보유하고 있는 유전자원에 대한 보존, 관리, 정보수집 및 유전자원 중복보존 등이 효율적으로 진행될 수 있도록 국가관리 기법을 개발한다. 이를 위해서 가축유전자원 관리지침을 개발하고 천재지변, 구제역, AI 등 다양한 환경변화로부터 유전자원의 안전보존을 위해 생축, 생식세포(정액, 수정란 등) 동결보존 등 중복보존을 실시한다. 이와 연계하여 우리나라의 대표

농업유전자원의 보존 • 관리 및 이용에 관한 법률 제16조에 의거 가축유전자원에 대한 책임기관(국립축산과학원 가축유전자원시험장)과 관리기관(11개소)이 지정·운영으로 각 관리기관에 보유하고 있는 유전자원에 대한 보존, 관리, 정보수집 및 유전자원 중복보존 등이 효율적으로 진행될 수 있도록 국가관리 기법을 개발한다. 이를 위해서 가축유전자원 관리지침을 개발하고 천재지변, 구제역, AI 등 다양한 환경변화로부터 유전자원의 안전보존을 위해 생축, 생식세포(정액, 수정란 등) 동결보존 등 중복보존을 실시한다. 이와 연계하여 우리나라의 대표적인 재래종인 재래소(칡소, 흑우), 재래흑염소, 재래돼지 및 재래닭에 대한 시료를 수집하고, mtDNA , MS 표지 등 FA O에서 권고하는 특성평가 마커시스템을 이용한 분자생물학적 특성평가를 실시하여 각 축종별, 품종별 기원 및 유전적 특성을 파악하여 가축유전자원의 보존·관리에 적용하고 나아가 국제적 대응을 위한 과학적 근거를 확보한다.
장기건조표본에 대한 DNA 분석 성공률을 개선하기 위해서 딱정벌레목 및 나비목의 장기건조표본을 대상으로 1∼15년, 15∼30년, 30∼60년 등 보존 기간에 따른 연차별 표본에서 효과적인 DNA 추출법과 손상 DNA로부터 DNA 바코드 분석을 원활히 하고자, 개체별 보존 상태 및 기간에 따른 DNA 단편화를 측정하여 이에 맞는 프라이머 세트를 재구성해 최종적으로 총 120종의 장기건조표본을 대상으로 DNA 바코드 분석 성공률을 개선한다.
농업곤충의 신속하고 정확한 종 동정을 위하여 우리나라 농업곤충 및 외국산표본 확보 및 G-DNA를 추출한다. 추출된 G-DNA는 초저온냉동고에 보관하고 샘플의 정보는 MS-Excel을 이용하여 DB화 한다. 기본 분자 마커인 LCO1490+HCO2198 프라이머 세트 혹은 특이 분자마커를 이용하여 증폭 후 염기서열을 판독, 확보한다. 확보된 서열들에 대해서 염기서열 분석 프로그램인 MEGA 5를 이용하여 각 종별 유전자 특성을 분석한다.
난분류 농업 해충 총 80종(3년간)에 대해 유(약)충 및 성충 채집 및 이에 대한 유, 성충의 DNA 바코드 영역 분석, 유, 성충 염기서열 비교분석, GenBank 정보와 비교분석, 해당 유, 성충의 사진 정보 축적 등을 통한 조기 종 동정 정보 축적을 통한 조기 종분류 기반 구축과 지역변이 가능 곤충 10종에 대한 최소 2개 지역 시료를 채집하여 DNA 분석을 통해 유전적 다양성, 지역적 유전변이 및 지역적 유전적 관련을 비교·분석한다. 이를 통해 선발종에 대해서 COI영역 및 ITS2 영역의 분석을 통해 집단유전학적 구조 및 계통분석을 수행한다.
누에 유전자원의 안전 계대보존을 위해 국내 종자은행에 등록되어 있는 누에 340 품종에 대한 분자생물학적 특성분석을 통한 품종특성 파악, 형질 정보 축적, 판별마커 개발 및 외부 유전자원 도입 등을 추진한다. 또한 뽕나무 및 오디의 특성조사를 통한 보존·관리기술을 확립한다.

Abstract ▼

National Institute of Animal Science(NIAS) appointed a total of eleven institutes to our partners responsible for livestock resource management. They are composed of nine livestock research institutes of nine provincial governments and two universities. These partners hold a total of 18,097 animals

National Institute of Animal Science(NIAS) appointed a total of eleven institutes to our partners responsible for livestock resource management. They are composed of nine livestock research institutes of nine provincial governments and two universities. These partners hold a total of 18,097 animals representing 27 lines of 11 breeds of 5 livestock species. We also have partners of private chicken farms raising around 400 chicks of four indigenous native lines(aliases of these are Hyonin, Poongdong, and Hwengsong Herbal chick). In our experiment station, we constructed a chicken house to make duplicate preservation of native chicks of NIAS (' 09.7.22.). Animal Genetic Resource Information Management System(AGRIMS) is developed to collect and manages information of our livestock genetic resources generated by us and our partners through web interface.(http://www.angr.go.kr/agrims/) Data from six livestock species collected through AGRIMS system were amounted to 8,000 in late 2011. In 2010, we published a brochu re explaining management of genetic materials, data collection,characterization of animals, and related laws or regulations. We made various efforts-workshops, symposiums and discussed the importance of genetic resource management and propagation of indigenous livestock species and breeds.
The bloods from 10 cattle populations, 9 goat populations, 4 pig populations, and 15 chicken populations, were obtained from NIAS, our 11 parteners, and several farms. Two types of genetic markers, mitochondrial(mtDNA) sequences and Mcrosatellite(MS) markers recommended from food and Agriculture Organization(FAO), were used to verified to the genetic diversity and phylogenetic status. In results of mtDNA analysis, the population-unique haplotypes were found from each species. Especially, in case of cattle, 46 of 65 haplotypes were the population-unique haplotypes. The native black pig were classified two subgroups, Asian and European types. The distribution pattern of these types was different among populations. In results of MS analysis, observed heterozygosity(Ho) was lower than expected heterozygosity(He) in several populations of each livestock. These results suggest that breeding program were performed by using limited male animals. In results of polymorphic information content(PIC), only 59(45%) MS mark ers of 130 are higly informative(PIC> 0.6) which make them useful in genetic diversity - 10 - studies. These results in this study might be used as basic information for planning of conservation and management of AnGRs. In addition, more informative MS marker will be select and apply for valuation of genetic diversity of AnGRs.
The DNA degradation of insects old specimens has been limited full-length (658 bp) sequencing. The challenges associated with the retrieval and authentication of degraded DNA extracts from old specimes were principally limited to analyze the relatively short sequences (< 300 bp). Furthermore, almost protocols in order to analyze the degraded DNA contained the cloning process after PCR causing the time-consuming and the rising costs.To overcome these problematic circumstances, we tried a modified method to analyze full-length of DNA barcoding region applied in old insects specimens(< 60 year-old) passing through three steps for the every 15 year-preserved periods consecutively. As the results, first, in 1∼15 year-old specimens, the universal primers were proved to be useless by low rates of PCR success (0∼10.52%). But when annealing temperature selectively modified between 45 to 52℃, it could be improved the success rate of PCR up to 25∼ 42.8%. However, the most effective improvement was to apply the newly des ired specific primers, which targeted identical region of universal primers. This method had increased the PCR success from 24.5% to 84% in Elateridae. Second, in 15∼30 year-old specimens, the success rates of PCR and sequencing were lower than 1∼15 year-old specimens, it is indicated that DNA degradation might be intensified over time. The most effective success rates showed in the application of the specific primers designed under 480 bp of target region in family and genus levels. This method was useful to raise the success rates of PCR and sequencing up to 84%. Third, in 30∼60 year-old specimens, DNA degradation of these samples was extremely progressed under 300bp length. To resolve this circumstances, a direct sequencing after PCR with species-specific overlapping primer sets per each species was employed. All of 28 species have been successfully analyzed although 178 samples (79%) are completely generated barcoding sequences ranged from 640 to 658 bp and 47 samples (21%) are partially sequenced ranged from 100 to 500 bp. Thus, the result showed that the direct PCR sequencing using the overlapping primer sets per species appears to have great potential efficiency for analysis of degraded DNA without incorrect sequences.
In the results of DNA barcoding on the Agricultural insects, the DNA stocks on total 5,185 individuals, 1,468 species, 68 families, five orders were producted, and the DNA barcodes on 3,101 individuals, 1,197 species, 63 families, five orders were obtained. The main results of DNA barcoding on the Agricultural insects are as followings: ' Construction of the molecular taxonomic system on 202 South Korean native butterfly species' , - 11 - ' Construction of DNA barcode library ― Natural enemy resources: 27 species of the South Korean Cantharidae, 38 species of the Far East Asian Coccinellinae; Agricultural pest insects: 56 species of the South Korean Chrysomelidae; 367 species, six families of the South Korean moth' . The obtained DNA barcode will be databased for the construction on the molecular taxonomic system of the agricultural insects.
For DNA sequence-based matching techniques for associating immatures and adults, a total of 80 species of Noctuidae and Pyralidae belonging mainly to agricultural pests were collected and their Barcode sequence were analysed. All species analyzed revealed identical 658 bp, composed of 219 amino acids. Sequence divergence between adult and immature stages ranged from 0 bp ～ 16 bp, indicating some species are very high in their sequence divergence. Along with the sequence information picture of all species were taken for both immature and adult stages for future reference. Screening of geographic variation of ten insect species provided the potential to have sequence divergence in some species, requiring further detailed investigation. Thus, two pest species, spodoptera litura and Cnaphalocrocis medinalis, were chosen and analyzed for COI and ITS2 from each a total of 159 and 187 individuals, respectively, collected from six Korean and five Chinese localities and eight Korean and five Chinese localities, respe ctively. The analysed sequences were subjected to several population genetic analysis and found a high genetic relationships of Korean populations to Chinese populations, along with several interesting features.
Genetic resources are current and future ground for evolution of new properties and,thus, they should be well organized and kept with systematic manner, because they may eventually provide ground for the production of new properties kept in the genetic resources. Once lost, it would be not possible to recover permanently. Fortunately, more than 3000 silkworm (Bombyx mori) strains are still maintained in Europe and Asia and these carry genetic differences. In the Republic of Korea, more than 340 silkworm strains remain under conservation in a government institute. These strains are annually reared, and scores from indoor rearing are analyzed for consistent character maintenance. Thus, individuals with unstable heritable characters are discarded for better keep pure lines. Nevertheless, still much confusion on the genetic stock exists.
Such abundance reversely may indicate the difficulty and complexity to discriminate one strain from the others. Furthermore, recent advance in molecular techniques again requires reexamination of once well established strains, because new insight into previous strains often results in obscurity.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 4
• SUMMARY ... 9
• 제1장 서 론 ... 12
• 제1절 연구의 필요성 및 범위 ... 12
• 제2절 연구의 목적 ... 14
• 제2장 국내외 기술개발 현황 ... 15
• 제1절 국내 관련기술 개발 현황 ... 15
• 제2절 국외 관련기술 개발 현황 ... 15
• 제3절 국내ㆍ외 기술개발현황에서 차지하는 위치 ... 16
• 제3장 연구개발수행 내용 및 결과 ... 18
• 제1절 연구개발 범위 ... 18
• 제2절 연구개발 방법 ... 19
• 제3절 연구 결과 ... 35
• 제4장 연구개발목표 달성도 및 대외기여도 ... 236
• 제1절 목표대비 달성도 ... 236
• 제2절 정량적 성과 ... 237
• 제5장 연구개발결과의 활용계획 ... 241
• 제1절 추가연구의 필요성 ... 241
• 제2절 활용계획 ... 241
• 제6장 중요 변동사항 ... 243