초록 ▼

1. 개다시마 군락지 연구
개다시마의 서식지 연구를 위하여 강원도 고성군 일대와 강릉시 사근진 해역 주변에서 청취조사 및 잠수조사를 실시하였으며, 조사 방법으로는 어촌계원, 잠수부, 해녀 등 각 해역의 토착민을 대상으로 수산생물 특성과 생산량조사, 개다시마의 서식밀도 등을 조사하였다. 또한 전문 잠수부를 이용하여 수중카메라, 수중비디오로 수중생태계를 촬영하였으며, line transect 방법으로 수직별 해조류 출현종과 $10{\times}10cm$로 세분된 $50{\times}50cm$

1. 개다시마 군락지 연구
개다시마의 서식지 연구를 위하여 강원도 고성군 일대와 강릉시 사근진 해역 주변에서 청취조사 및 잠수조사를 실시하였으며, 조사 방법으로는 어촌계원, 잠수부, 해녀 등 각 해역의 토착민을 대상으로 수산생물 특성과 생산량조사, 개다시마의 서식밀도 등을 조사하였다. 또한 전문 잠수부를 이용하여 수중카메라, 수중비디오로 수중생태계를 촬영하였으며, line transect 방법으로 수직별 해조류 출현종과 $10{\times}10cm$로 세분된 $50{\times}50cm$의 크기의 방형구를 이용하여 해조 출현종을 조사하였다. 현존량은 방형구내의 해조류 전량을 채취하여 종별 생중량을 측정하였다. 조사의 정확성을 기하기 위하여 수중 카메라와 수중비디오로 수중생태계를 촬영하여 실내에서 비교 분석하였다.
2. 생활사 및 생태 연구
개다시마 최대군락지인 강릉시 사근진 해역 주변에 서식하고 있는 개다시마의 생장과 성숙 등을 월별 정기적으로 생태 조사하였다. 이를 위하여 매월 엽체의 엽장, 엽폭 등 생장을 조사하였고, 조직절단을 통한 영양세포의 변화를 비롯하여 현미경으로 성숙 자낭반 부위를 절단하여 검경하였다.

Abstract ▼

Ⅱ. Objective
Many seaweeds exhibit distinctive seasonal growth phases; winter growth phase and summer degeneration phase. In the marine ecosystem, seaweeds provide valuable fishery resources such as fishes, abalones and other invertebrates with habitats and spawning grounds. They can be valued pr

Ⅱ. Objective
Many seaweeds exhibit distinctive seasonal growth phases; winter growth phase and summer degeneration phase. In the marine ecosystem, seaweeds provide valuable fishery resources such as fishes, abalones and other invertebrates with habitats and spawning grounds. They can be valued products for human as well as for marine invertebrates.
Aquaculture is one of the promising activities for seaweed production. Activities of seaweed aquaculture have pretty much short history, but they have been intensified for last decades.
The submarine forest is very important in the structure and function of the subtidal ecosystem as is the terrestrial forest. The marine plants that form submarine forest are of several benthic marine algae belonging to Laminariales. Recently, the submarine forest and its composing species has increasingly drawn a public attention because of its ecological function in the marine ecosystem. Moreover, the recovery of coastal fisheries resource is achieved by the construction of seaweed forest, that, in turn, could promote the quality and quantity of coastal fisheries resource and improve the structure of coastal ecosystem.
With an aid of phycological researches including taxonomy, systematics, morphology, ecology, physiology, genetics, and breeding, the aquaculture business have rapidly developed. Kjellmaniella crassifolia Miyabe has been one of the valuable seaweed crops cultured in the Korean costal waters of the East Sea. Recent changes in environmental parameters, however, have adversely influenced the crop production, thereby stimulating a technology development for the species restoration.
We studied the ecology and gamete preservation of Kjellmaniella crassifolia inhabiting Gangneung coasts for laboratory production. The production technology will be applied to aquaculture and wild stock restoration of the species. Mass production of the species will be used for human consumption and extracted for some functional matters.
Ⅲ. Contents and scopes of the study
1. Survey on the life cycle and ecology
Kjellmaniella crassifolia Miyabe is perennial species and endemic to East Sea of Korean Peninsula. The species inhabits deep water of the sea. For a biological information, growth of the species have been monitored for a year. Materials of this study were monthly collected from deep water (depth, 20~30m) of Gangneung coast by scuba diving. We monthly investigated growing pattern, maturing time, standing crop and distribution. Maximum blade length, 110 ± 45㎝, was gained in July, thereafter releasing zoospore in November. Sporangial sori started to form from September, reaching 85% maturity in November and 89% in December.
Kjellmaniella crassifolia was abound in Gangneung coasts by 1990s covering about 50ha at density of 500 ind/10m². In our 2004 survey the habitats reduced to about 2ha with a density of 1-2 ind/10m². The habitats and density increased to 7ha and 5-10 ind/10m², respectively, after our effort to restore wild population of the species. Kjellmaniella crassifolia was dominant species by 1990s at water depth of 20-30m. However, the habitats were dominated by Agarum clathratum with a coverage of 80-100%. Ratios of blade length and width were 2.8~2.9 for maturing Agarum clathratum and 4.1 for growth phase of the species.
2. Development of seed production
For the determination of optimum cultivation conditions, the sporophyte was grown under 5 temperatures (5, 10, 15, 20, 25 and 30℃), 5 illumination quantities (10, 20, 40, 60, and 80 μ㏖․m^-2․s^-1), and 5 photoperiods (5:19, 8:16, 12:12, 16:8, and 19:5) in a medium PESI for 40 days. Zoospore germination was strongly influenced by higher temperatures: all the zoospores failed to germinate in the temperatures higher than 20℃. Best zoospore germination rate (80%) was achieved in the conditions of temperature 15℃ and illumination 40 μ㏖․m^-2․s^-1, while worst (10%) was in the conditions of temperature 20℃ and illumination 80 μ㏖․m^-2․s^-1. Optimum condition for gametophyte growth was 10℃ and illumination 80 μ㏖․m^-2․s^-1. In the condition the gametophytes grew to 286.0±35.20 ㎛ for the experiment. The favored photoperiod (growth) for sporophyte was 8L:16D (530±120㎛) followed by 12L:12D (410±112 ㎛), 16L:8D (290±35㎛), 5L:19D (310±52 ㎛), and 19L:5D (180±31 ㎛). The seeds produced were maintained in the hatchery for a month using free culture and transplantation in the rope and then transplanted into the open coasts.
3. Sequence analysis of Nuclear 18S rDNA from the seaweed Kjellmaniella crassifolia (Phaeophyta) in East coast of Korea
The polymerase chain reaction (PCR) catalyzed by Taq DNA polymerase is a very powerful and sensitive analytical technique, with applications in many diverse fields, including molecular biology and population genetics (Innis et al., 1990). Since Hong et al. (1992; 1995) established a simple method for the rapid extraction of nucleic acids from seaweed using lithium chloride, PCR has also been applied to the field of seaweed molecular biology. Molecular study of evolutionary divergences requires sequencing of very slowly evolving genes, such as nuclear-encoded genes for small and large subunit ribosomal RNA (SSU and LSU rDNA). SSU (18S rDNA) is more highly conserved than LSU.
Kjellmaniella crassifolia Miyabe, an edible brown alga inhabits in limited regions from Gangneung to Goseong of East Sea and is very a useful seaweed compared to other Laminaria sp. However, K. crassifolia was gradually declined by several effects, such as over-utilization and typhoon etc.
In this study, we investigated the phylogenetic diversity using the nuclear 18S ribosome RNA gene (18S rDNA) from the seaweed K. crassifolia collected from east coast of Korea. Also we aimed to obtain a new genetic marker for identification and to compare it to other Laminaria sp.
The results was Nuclear 18S ribosome RNA gene (18S rDNA) from the seaweed K. crassifolia collected from east coast of Korea was amplified using the polymerase chain reaction and its sequence was analysed. The 18S rDNA sequence was an 1838 bp and showed 97% homology to L. japonica (the GeneBank accession number AB022817.1). The Genetic markers to classification were obtained three candidated gene sequences; one gap sequence and two insert sequences.
4. Gametophyte Separation and Preservation for the Artificial Seed Production of K. crassifolia
Kjellmaniella crassifolia Miyabe, an edible brown alga inhabits in limited regions from Gangneung to Goseong of Eastern Coast of Korea. We conducted to reveal the conditions of separation and preservation of gametophyte for artificial seed production of K. crassifolia. The released zygotes were cultured on four different gelling agents with added Provasoli's enrichement (PESI) under different light conditions (20, 40, 60 and 80 μ㏖․m^-2․s^-1) and temperatures (5, 10, 15, 20 and 25℃) at 12L:12D. The highest percent of zygotes` germination (85%) was produced on semi-solid medium composed of 1.0% transfer gel agar under 15℃ and 20 μ㏖․m^-2․s^-1 after 4 weeks culture.
Zygotes germination was formed in PESI liquid medium at the percent of 93.5% under 20℃ and 20 μ㏖․m^-2․s^-1. Maximum growth of zygote was 252±19.7㎛ on 1.0% transfer gel agar under 15℃ and 40 μ㏖․m^-2․s^-1 after 5 weeks culture and was 76.7±2.8㎛ in PESI liquid medium under 20℃ and 40 μ㏖․m^-2․s^-1. The number of separated male and female gametophyte from germinated zygotes was 157, 93 on 1.0% transfer gel agar, respectively and 14, 28 in PESI liquid medium, respectively. Maximum growth of separated male and female gametophyte were 575±28.3 ㎛ under 5℃ and 60 μ㏖․m^-2․s^-1 and 686±35.4㎛ under 20℃ and 20 μ㏖․m^-2․s^-1 in PESI liquid medium for 3 weeks, respectively. The highest percentage of fertilized ratio was 93.3±5.8% under 15℃ and 20 μ㏖․m^-2․s^-1 in PESI liquid medium. These results show that the best condition for separation and preservation of gametophyte (male and female) was culture on 1.0% transfer gel agar under 15℃ and 20 μ㏖․m-2․s^-1. It would suggest that K. crassifolia could be possible to be mass-produced by cultivated female and male gametophytes if necessary any time during the year, and it is believed to be build a gene bank by preserving the male and female gametophytes of K. crassifolia.
5. Studes on optimal water depth for the aquaculture K. crassifolia Cultivation technology of K. crassifolia was first developed in Korea. In order to examine cultivation availability for K. crassifolia from northern coastal area (Gangneung) of the East Sea. The cultivation was done at 5, 10, 15, 20, and 30m. Unlikely our general knowledge that the species inhabits deeper coasts, shallow coasts were preferred for the species. The highest growth depth is 5m reached blade length 75.9±18.9cm. It is reverse result consider restoration area.
6. Studes on Kjellmaniella crassifolia for wildstock restoration K. crassifolia distributes in the coastal waters 20~30m depth and usually grows at 25~30m. We had K. crassifolia transplanted in the coastal waters where wild populations are decreasing due to enforced human activities. The techniques used for the restoration were based on the techniques developed by East Sea Fisheries Research Institute. To developing the techniques for the restoration, East Sea Fisheries Research Institute (ESFRI) in National Fisheries Research and Development Institute (NFRDI) set up the study field in Sagunjin, Gangnung. The transplanted seaweed, 320±32 ind/m², survived 15±5 ind/m², representing 4.7% survival rate.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 23
• CONTENTS ... 28
• 목차 ... 29
• 제 1 장 연구개발과제의 개요 ... 31
• 제 1 절 기술적 측면 ... 32
• 제 2 절 경제.산업적 측면 ... 32
• 제 3 절 사회.문화적 측면 ... 33
• 제 2 장 국내외 기술개발 현황 ... 34
• 제 3 장 연구개발수행 내용 및 결과 ... 35
• 제 1 절 개다시마 군락지 연구 ... 35
• 제 2 절 생활사 및 생태 연구 ... 54
• 1. 서식지 해양환경 특성 연구 ... 54
• 2. 생활사 연구 ... 61
• 제 3 절 유주자 방출 유도 연구 ... 66
• 제 4 절 환경조절에 의한 종묘생산연구 ... 67
• 1. 환경조절에 의한 실내 종묘생산 연구 ... 67
• 2. 자연 유수식 종묘생산 연구 ... 76
• 제 5 절 유전자 위치 및 분석 연구 ... 82
• 1. 시료채집 및 전처리 ... 82
• 2. DNA 추출 ... 82
• 3. DNA 정량 ... 83
• 4. PCR 증폭 ... 83
• 5. Agarose gel electrophoresis 및 DNA 회수 ... 83
• 6. DNA ligation 및 transformation ... 84
• 7. Plasmid 추출 및 제한효소처리 ... 84
• 8. 동해안 개다시마의 18S rDNA 염기서열분석 ... 84
• 9. 18S rDNA의 증폭 ... 84
• 10. DNA 염기서열의 비교 ... 85
• 제 6 절 암.수배우체 분리 및 보존 연구 ... 88
• 1. 채집 및 전처리 ... 88
• 2. 배지 ... 89
• 3. 유주자의 발아 및 배우체와 아포체의 생장 연구 ... 91
• 4. 배우체 분리 및 번식 연구 ... 103
• 5. 배우체 수정 연구 ... 107
• 6. 배우체 보존 연구 ... 108
• 제 7 절 적정수심 구명에 의한 양식기술개발 연구 ... 111
• 1. 월별 생장 연구 ... 111
• 제 8 절 서식지복원 연구 ... 114
• 1. 그물망에 의한 서식지복원 연구 ... 114
• 2. 해중림초 이식에 의한 서식지복원 연구 ... 118
• 제 9 절 개다시마 산업화에 따른 경영수지 분석 ... 123
• 1. 건강식품 개발에 따른 경영수지 분석 ... 123
• 제 4 장 참고문헌 ... 128
• 끝페이지 ... 133