초록 ▼

○ 총 5종의 해양 난균강 원생생물 균주와 숙주를 채집하고 분리 및 실내배양을 수행하였으며, 최적 배양 조건 구축 및 주요 숙주, 보조 숙주를 규명하였음. 균주 2종에 대한 감염특성을 규명하고 융합오믹스 테스트와 균주를 선별함
○ 해양 와편모조류 Prorocentrum minimum 공생미생물 특성 규명. 부착 및 자유유영 미생물중 Alteromonadales의 Marinobacter와 Rhodobacteraceae에 속하는 Seohaeicola 속이 매우 중요한 작용을 파악함. 원생생물-공생미생물간의 수평적유전자전달(HGT)

○ 총 5종의 해양 난균강 원생생물 균주와 숙주를 채집하고 분리 및 실내배양을 수행하였으며, 최적 배양 조건 구축 및 주요 숙주, 보조 숙주를 규명하였음. 균주 2종에 대한 감염특성을 규명하고 융합오믹스 테스트와 균주를 선별함
○ 해양 와편모조류 Prorocentrum minimum 공생미생물 특성 규명. 부착 및 자유유영 미생물중 Alteromonadales의 Marinobacter와 Rhodobacteraceae에 속하는 Seohaeicola 속이 매우 중요한 작용을 파악함. 원생생물-공생미생물간의 수평적유전자전달(HGT)이 일어난 cold shock protein, SAM-dependent methyltransferas, Metallophosphoesterase, Peroxiredoxin (PRDX) 유전자를 규명함
○ 우리나라의 연안환경에서 해양 원생생물 분리, 배양하여 총 15종의 신계통주 단종배양체를 확립함. 총 12종의 단종배양체에 대해 염기서열을 분석 및 계통수 제작을 통해 유전적 특성을 규명함. 외부의 형태, 세포 내 미세구조 등의 특성을 분석 및 신계통주 11종에 대한 영양방식 및 먹이종과 포식종을 규명함. 분자생물학적, 형태학적 분류 key에 따른해양원생생물 차세대 분류시스템 확립을 위한 분류 key 2개를 발굴함
○ 은편모류 기반 해양 원생생물 89종의 단종배양체 확립 (은편모류 15종, 와편모류 50종, 규조류 10종, 유글레나류 2종, 침편모류 1종, 녹조류 6종, 섬모류 4종)과 형태 및 분자생물학적 방법을 사용하여 분류함. 유용한 배양체의 대량배양 (와편모류 4종, 은편모류 1종)을 통한 독성물질, 소염진통 효능 등이 우수한 이차대사 물질 분리 및 구조 규명하였으며 이 중 활성이 우수한 신규 화합물 6개를 분리함

(출처 : 보고서 요약서 3p)

Abstract ▼

IV. Results
○ Regular collection activity was conducted and oomycete strains were collected from 6 sites of the East sea (two sites in Gangneung) and West sea (two sites in Seocheon) South sea (Jindo and Wando)
○ Investigated the host distribution and ecosystem from each region
○ The optima

IV. Results
○ Regular collection activity was conducted and oomycete strains were collected from 6 sites of the East sea (two sites in Gangneung) and West sea (two sites in Seocheon) South sea (Jindo and Wando)
○ Investigated the host distribution and ecosystem from each region
○ The optimal culture conditions of oomycetes were carried out using 10-20 ℃ incubator, 12:12 hr photoperiod and the alga culture medium. Pythium porphyrae was isolated and cultured alone. The infection experiment was conducted in Arasaki-B medium
○ Oomycete strains were maintained by storing at -78ºC with 1M sucrose, 5% proline, 5% glycerol and KNU-Cryo1 (medium: bovine serum: glycerol = 6:3:1) for cryopreservation
○ Oomycete strain was identified by molecular phylogeny analysis
○ The infection characteristics and life cycle of Pythium porphyrae was investigated. The possibility that Pythium porphyrae was originated from the land was confirmed
○ The infection characteristics of Olpidiopsis porphyrae var. koreanae was investigated by the form and physiological analysis through TEM
○ Collected oomycete strains with hosts were selected for Integrated-omics analysis and DNA, Total RNA and protein were isolated and analyzed
○ By considering national needs for scientific, industrial, economical, and ecological aspects, research speices were selected, and these include the diatoms Ditylum brightwellii and Cyclotella meneghiniana, green algae Tetraselmis suecica, and dinoflagellate Prorocentrum minimum and Amphidinium carterae.
○ Analyze culture characteristics of diatom, green algae, and dinoflagellagte according to 10~30°C, and 10~40‰, respectively.
○ Determination of toxicity EC50 to research species exposed to environmental toxicants (Cu, Ni, Cd, Pb, Endosulfan, Metolachlor, PCB (Aroclor 1016), Bispenol A, etc.)
○ RNA isolation, and 302 cDNA construction
○ Determine transcriptomes of diatom Ditylum brightwellii using GS-FLX, and identify gene information (334,592 reads, 149 Mb). GO analysis and determination of 6,162 unique genes.
○ Determine 173,600,654 reads of transcriptomes (17.5 Gb) from diatom Cyclotella meneghiniana. GO analysis and determination of 44,060 unique genes.
○ Determine 740,557 reads of transcriptomes (290 Mb) from green algae Tetraselmis suecica.
○ Determine 610,301 reads of transcriptomes (245 Mb) and 11,933 unique genes from Prorocentrum minimum.
○ Determine 120,137,900 reads of transcriptomes (12.1 Gb) and 39,780 unique genes from Amphidinium carterae.
○ Gene cloning of functional genes (HSPs, P450, SOD, GST) from diatoms, green algae, and dinoflagellates, gene expression analysis, and molecular phylogenetic analysis.
○ Determine free-living and particle-associated bacteria to marine diatom Cyclotella meneghiniana. Reveal Persicirhabdus and Oceanicaulis for free-living, and Marinobacter for particle-associated bacteria, respectively.
○ Determine free-living and particle-associated bacteria to marine green algae Tetraselmis suecica. Reveal Alteromonadales belonging Marinobacter for free-living, and Marinobacter for particle-associated bacteria.
○ Determine free-living and particle-associated bacteria to marine dinoflagellate Prorocentrum minimum. Reveal Alteromonadales belonging Marinobacter and Rhodobacteraceae belonging Seohaeicolafor.
○ Isolation of bacteria associated to P. minimum culture stages. Determine Alteromonas sp., Marinobacter sp. using 16S rDNA analysis.
○ Identify horizontal gene transfer (HGT) genes, including cold shock protein, SAM-dependent methyltransferas, Metallophosphoesterase, and Peroxiredoxin (PRDX).
○ For the conduction of clonal cultures, 15 strains were settled. For genetic analysis, 12 strains were investigated.
○ Ecological characterizations were investigated from 11 strains, and physiological characterizations from 12 strains.
○ Two taxonomical keys were developed as planned.
○ Established 80 uni-protist cultures of the cryptophyte-mediated key marine protist species (10 benthic or planktonic cryptophytes, 50 benthic or planktonic dinoflagellates, 2 euglenoids, 10 diatoms, 6 chlorophytes, 1 raphidophyte, 4 ciliates).
○ Each of them was set up as laboratory strain, and also used for taxonomic works by morphological and molecular genetic studies. In addition, small scale cultivation of each of the strains (35 dinoflagellates, 3 euglenoids, 3 cryptophytes, 2 cyanobacteria, 2 chlorophytes) was performed for the screening of useful strains with bioactive applicabilities.
○ Among the selected strains with useful bioactivities 7 dinoflagellates and 1 cryptophyte strains were mass cultured to isolate the secondary metabolite molecules showing cytotoxicity, antioxidant, and anti-inflammatory effect, to be followed by the exploration of the molecular structures.

(출처 : SUMMARY 10p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 3
• 요 약 문 ... 4
• S U M M A R Y ... 9
• C O N T E N T S ... 14
• 목차 ... 15
• 제 1 장 연구개발과제의 개요 ... 16
• 제 1 절 연구개발 개요 및 목적 ... 16
• 제 2 절 연구개발의 필요성 ... 21
• 제 3 절 연구 목표 및 연구개발의 범위 ... 24
• 제 2 장 국내외 기술개발 현황 ... 27
• 제 3 장 연구개발수행 내용 및 결과 ... 31
• 제 1 절 세부과제별 주요 연구내용 ... 31
• 제 4 장 목표달성도 및 관련분야 기여도 ... 64
• 제 1 절 연구개발 최종목표, 연차별 연구개발 목표 및 내용 ... 64
• 제 2 절 연구개발 목표 및 달성도 ... 71
• 제 3 절 관련분야 기술 발전 기여도 ... 76
• 제 5 장 연구개발성과의 활용계획 ... 79
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 80
• 제 7 장 참고문헌 ... 81
• 끝페이지 ... 86