초록 ▼

■ 주요 해양생물 DNA바코드 분석과 분자마커 활용 종판별 기술 개발 및 현장 적용과 평가
- 한국해역에 서식하는 주요 해양생물 1050종의 DNA바코드를 확보(어류 317종, 연체동물 210종, 절지동물 147종, 극피동물 373).
- 적조발생의 원인이 되는 미세조류 및 해파리 종목록을 작성 후 적조의 원인이 되는 식물플랑크톤 27종을 배양 유지하고, COI 염기서열을 분석.
- 주요 간극생물 중 선충류 40종, 저서성 요각류 5종의 18S rDNA 바코드 확보.
- 분자마커를 활용한 760여종의 종판별용

■ 주요 해양생물 DNA바코드 분석과 분자마커 활용 종판별 기술 개발 및 현장 적용과 평가
- 한국해역에 서식하는 주요 해양생물 1050종의 DNA바코드를 확보(어류 317종, 연체동물 210종, 절지동물 147종, 극피동물 373).
- 적조발생의 원인이 되는 미세조류 및 해파리 종목록을 작성 후 적조의 원인이 되는 식물플랑크톤 27종을 배양 유지하고, COI 염기서열을 분석.
- 주요 간극생물 중 선충류 40종, 저서성 요각류 5종의 18S rDNA 바코드 확보.
- 분자마커를 활용한 760여종의 종판별용 종 특이적 프로브/프라이머를 설계 및 21종은 Realtime PCR을 통해 종판별 프로브/프라이머의 정확성 검증.
- 해양주요 생물 종판별을 위한 DNA칩(어류 45종, 무척추생물 41종, 해양 미세조류 36종, 해파리 8종)의 종 특이적 프로브 제작 및 성능 평가.
- 차세대 유전자분석 기술을 적용하여 각 해역의 어란, 무척추동물 유생, 동물플랑크톤의 종조성 분석.
- 어류의 산란기와 자치어의 총 출현량 자료를 확보 및 어란을 이용한 산란장 탐지에 이용될 DNA칩 제작을 위해 미토콘드리아 DNA 16S와 CO1 유전자 염기서열 분석.
- 생물다양성 연구에 매우 중요한 해양 간극생물의 e-boucher 확보 및 슬라이드 표본 제작.
- 현장분석용 DNA칩 분석시스템 장비 제작에 필요한 각 컴포넌트 기기들의 장단점을 분석하고, 가상의 장비 모형을 제시.

■ 유해생물 음향탐지 분야
- 초음파의 후방 산란특성 이론을 기반으로 적조 피해 유발 종인 Cochlodinium polykrikoides의 탐지를 위한 통합 음향시스템 개발. 시스템은 실시간 적조생물 탐지를 위한 음향 탐지부와 제어 및 운용을 위한 원격 제어부로 구성. 유선/무선네트워크를 통해 육상 기지국으로 음향자료를 실시간 전송. 또한 원격 운용과 현장 운용이 가능한 형태로 개발하여 부이를 이용한 장기 관측 및 선박을 이용한 공간 분포의 파악이 가능한 형태로 구성.
- 제작된 적조 음향탐지시스템은 실험실 및 실해역 조건에서 반복적인 성능 평가를 통해 안정성 및 실효성을 검증. 본 연구를 통해서 남해안 적조 발생을 실시간으로 탐지할 수 있는 성능 및 현장 운용 가능성 제시.

■ 유용/유해 생물 유비쿼터스 정보서비스시스템 구축 및 국제 네트워크화
- 총 208개 참고문헌을 조사 하여 총 69,319건의 출현정보를 발굴, 수집하였으며 수집 자료를 분석하여 DB를 설계. 이를 근거로 Oracle 11g 기반의 DB를 구축하고 웹상으로 정보를 제공할 수 있는 시스템을 구축.
- 기 수집·정리된 자료를 기반으로 종별, 공간별, 시․공간적 분포패턴을 정량화하고 분석패턴을 통계 처리하여 GIS 기반의 출현정보 비교/분석 기능을 구현하였으며 과거 수온/염분자료를 통계 처리하여 해양물리 환경정보 DB를 구축.
- 무선통신환경에서 스마트폰을 통해 해양생물지리정보 및 통계처리정보를 검색, 참조할 수 있는 모바일 웹사이트와 모바일 앱 개발. 현장정보입력기능 구현을 통해 음향탐지분석정보를 실시간으로 수집하여 제공하는 유비쿼터스 정보서비스 구현.
- 국제 OBIS에 연계하기 위해, 기 구축한 DB에서 활용 가능한 자료를 추출하였으며, OBIS용 국내 DB 설계하고 자료를 이용하여 MS Jet Engine 기반의 DB를 구축하여 약 3,100 건의 KOBIS DB의 자료를 국제 OBIS 포털을 통해 검색 가능하도록 함. 국제 OBIS의 지역노드(Regional OBIS Node)인 KOBIS의 웹사이트(http://kobis.kiost.ac)를 구축.

(출처 : 초록 5p)

Abstract ▼

Ⅳ. Result
A. DNA barcode analysis of major marine organism and the development of species identification technique and in-situ analysis and assessment.
(1) Invertebrate
- Total 730 molecular markers, as the reference data for the design of species specific primer set were obtained and about

Ⅳ. Result
A. DNA barcode analysis of major marine organism and the development of species identification technique and in-situ analysis and assessment.
(1) Invertebrate
- Total 730 molecular markers, as the reference data for the design of species specific primer set were obtained and about 500 species specific probe/primer sets were designed.
- As a result of NGS(next generation sequencing) of mixed netted sample, there are comprised total above 42 species including arthriopoda 32 sp., mollusca 4 sp., echinoderm 2sp., etc. in tongyeong. In Ieodo, including Annelida 1sp, arthropoda 4sp, mollusca 1sp, cnidaria 1sp, chaetognatha 1sp, etc. total above 26 species are inhabited. In Gageocho, polychaete 6sp., arthropoda 2sp., chaetognatha 1sp, etc. total 9 species are inhabited.
- Species specific probe/primer sets were synthesized and total 11 probe/primer sets were validated for the species identification in the field.
- The major useful invertebrates (86 species) inhabiting soft-bottom of the southern and western coast of Korea were selected and collected during research period. Marine nematodes and benthic harpacticoids were analysed based on the 18S rRNA gene sequences to distinguish closely related species among them. Also, DNA sequences and digital images were obtained for constructing the database on the meiobenthic animals.
- Cross validation was performed between results from the NGS and the Real-time PCR(species specific primer) using the mixed sample in Tongyeong, Ieodo, Gageocho and Jejudo sites.
- Forty-one species specific DNAchips of invertebrates were developed and validated.

(2) Fishes (eggs, larvae, and adults)
- Total 317 molecular markers, as the reference data for the design of species specific primer set were obtained and about 230 species specific probe/primer sets were designed.
- Species specific probe/primer sets were synthesized and total 10 probe/primer sets were validated for the species identification in the field.
- As a result of NGS(next generation sequencing) of mixed netted sample, there are comprised total above 6 species including a wrasse and an anchovy. In Ieodo, 3 species were inhabited and In Gageocho, there's no report.
- Using morphological identification, fish eggs were identified to one species, Engraulis japonicus . and fish larvae were classified more than 30 species. The main spawning time were on July. The fish eggs and larvae of E. japonicus were dominated. The pattern of spawning time of major species such as E. japonicus and Leiognathus nuchalis was one peak in 2008 and two peaks in 2009. This was due to the difference of collection period, each week(2009) and two weeks(2008).
- In order to analyze the species composition of massive mixed fish eggs, the condition of NGS method was required to analyze 300 specimens and to obtain more than 300 sequences each sample(station) reads. Detection of small amount of DNA in NGS method was testified by comparing the species composition from NGS to those from Sanger library consisted of known species composition. By NGS, fish eggs were identified to 18 taxa. From the information of species distribution of fish eggs, it was elucidated that fishes spawned having the time difference within same spawning area and used spawning areas widely or narrowly depending on species, although their spawning time was similar.
- Portable DNA analysis tool kit was developed and applied in the field.
- Forty-five species specific DNA chips of invertebrates were developed and validated.

(3) Microalgae and jellyfish
- Development of DNA chip for jellyfish species isolation
· DNA barcoding analysis of 6 species of jellyfish in korea
· Selection of jellyfish probe and species isolation thus DNA chip
- Development of DNA chip for microalgae species isolation
· DNA barcoding analysis of 25 species of microalgae in korea
· Selection of microalgae probe and species isolation thus DNA chip
- Field application and development of microalgae species isolation technology using PCR method
· Obtained samples of 17 microalgaes in Tongyeong
· The barcoding analysis and microalgae species isolation by SEM
· Manufacture of species specific primer and field application
- Automatic species isolation study by FISH method
· Detection of Heterocapsa circularisquama by FISH method
· Establishment of antomatic species isolation technology

B. Studies on acoustical characteristics of harmful organism
- Acoustic target-strength (TS) measurements are presented for Aurelia aurita, Nemopilema nomurai, Cyanea nozakii, Dactylometra quinquecirrha and Dactylometra quinquecirrha in the coast of the Southern sea, Korea, 2008-2009. using active acoustic system in cage. As a result of studies, Acoustical target-strength (TS) of the jellyfish is distinct from the fishes and zooplanktons.
- The volume backscattering strength from the cultured media with Cochlodinium polykrikoides was calculated from the acoustic signals, and the interrelation between the number of cells and the acoustic signals was studied in the laboratory.
- Real-time acoustic system was developed to detect HABs, based on the backscattering properties of the biological target from the ultrasonic sound. For the purpose, integrated acoustic system was developed through technical evaluation of the each component, and performance evaluations of the system were carried out from laboratory and field test.
- The developed acoustic system was well worked with aspect of the function and performance under the independent and integrated condition. The integrated acoustic system was developed with two modes; remotely operation with buoy system and field operation with monitoring ship-type system.
- Confirmed to detect HABs. Additionally, comparing with the research equipment, the developed system has a considerably good performance, henceforth, can be modified or improved for specific purpose at any time.
- Although the developed acoustic system from this study must be technical modified, totally the system was proposed with to real-time detect HABs in the field condition. In the future, the system could be applied to make system implementation for monitoring HABs at coastal waters around Southern Sea, Korea.

C. Establishment of an biogeographic information system for useful/harmful marine organisms and participation in the international OBIS network
- Total 69,319 informations were collected from the journal of the Korea Society of Oceanography, the journal of the Korean Fisheries Society, Ocean and Polar Research and the reporters of the ocean projects.
- New Database system which can manage species list, taxonomy information, geographic information of marine organisms was designed and established using Oracle 11g.
- An ubiquitous information providing system which can search and retrieve biogeographic information from the database system was established.
- An acoustic detection data service system that collected in real time was established.
- A Field data insert module for tablet PC was developed.
- Data for KOBIS DB were retrieved from the Korea Marine Biodiversity Information System and quality controlled.
- New KOBIS DB system was developed and informations were inputted to it.
- The DiGIR software package was installed to link the KOBIS DB with the OBIS network and KOBIS data can be retrieved from the international OBIS portal.
- New KOBIS web-site was set up to provide KOBIS status and the data of the KOBIS DB.

(출처 : SUMMARY 19p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 3
• 보고서 초록 ... 5
• 요 약 문 ... 7
• SUMMARY ... 15
• 목차 ... 24
• CONTENTS ... 25
• List of Tables ... 26
• List of Figures ... 29
• 제 1 장 서 론 ... 39
• 제 1 절 연구개발의 필요성 ... 41
• 제 2 절 연구개발 목표 및 내용 ... 44
• 제 2 장 국내외 기술개발 현황 ... 53
• 제 1 절 국내 연구 동향 ... 55
• 제 2 절 국외 연구 동향 ... 56
• 제 3 절 현재까지의 연구개발 현황 ... 57
• 제 4 절 연구 수행 내용 및 방법 ... 59
• 제 3 장 연구개발 수행 내용 및 결과 ... 65
• 제 1 절 주요 해양생물 DNA바코드 분석과 분자마커 활용 종판별 기술 개발 및 현장 적용과 평가 ... 67
• 제 2 절 유해 생물 탐지 음향시스템 구축과 운용 기술 ... 224
• 제 3 절 주요 유용/유해생물 지리정보 DB구축, 국제 네트워크화 ... 361
• 제 4 장 목표달성도 및 대외기여도 ... 381
• 제 1 절 목표 달성도 ... 382
• 제 2 절 대외 기여도 ... 384
• 제 5 장 연구개발 결과의 활용계획 ... 386
• 제 6 장 참고문헌 ... 390
• 제 7 장 부 록 ... 400
• 끝페이지 ... 419