동해안 개다시마 Kjellmaniella crassifolia Miyabe (Phaeophyta) 종묘생산을 위한 배우체 분리 및 보존
Gametophyte Separation and Preservation for the Artificial Seed Production of Kjellmaniella crassifolia Miyabe (Phaeophyta) in East Coast of Korea
- 발행기관 강릉대학교 산업대학원
- 지도교수 진형주
- 발행년도 2007
- 학위수여년월 2007. 2
- 학위명 석사
- 학과 및 전공 해양생명공학과
- 원문페이지 vii, 59 p.
- 본문언어 한국어
초록/요약
In order to examine the separation and preservation of gametophyte for the seed production of Kjellmaniella crassifolia in large quantity, a native species in East Sea that has been in danger of extinction, the seedlings matured from the sea areas of Bongpo-ri, Goseong-gun, Gangwon-do were collected and the growth, maturity of gametophyte in liquid medium, solid medium and germination of zoospore and the growth of sporophyte were investigated. In addition, after dividing the gametophytes by medium in the formation phase of female and male gametophytes and homogenizing them, the propagating conditions of female and male gametophytes were examined. Then, by fertilizing each of female & male gametophytes in the liquid medium that had the best growth and maturation and their fertilization rate was measured and the experiments of preserving gametophytes and species by temperature, intensity of radiation and medium were conducted. The germination of zoospores was processed in 5 stages of water temperature (5, 10, 15, 20 and 25℃) and four stages of intensity of radiation (20, 40, 60 and 80 μ㏖·m^(-2)·s^(-1)). After cultivating them by liquid and solid medium, the highest cultivating rate was obtained at 20℃ of water temperature and 20 μ㏖·m-2·s-1 of intensity of radiation in liquid medium, but most of them did not germinate up to a month after cultivation and were in a state of zoospore in solid medium; therefore, it has been found that the germination was constrained. Like the germination of zoospores, the growth and maturation of gametophytes was processed in 5 stages of water temperature (5, 10, 15, 20 and 25℃) and four stages of intensity of radiation (20, 40, 60 and 80 μ㏖·m^(-2)·s^(-1)). After cultivating them by liquid and solid medium, the highest growth and maturation were obtained at 20℃ of water temperature and 40 μ㏖·m^(-2)·s^(-1) of radiation intensity. In addition, the better growth was found in low radiation intensity than in high radiation intensity, and it has been thought that they were benthic sea algae mainly inhabited in deep water and more affected by low radiation intensity. In terms of the separation of gametophytes, in case of agar, purified agar and high gel among solid media, there were difficulties in lifting the medium part that had zoospores at the time of separating gametophytes; therefore, it was conducted only in transfer gel among liquid and solid medium and especially, the separation was conducted a lot better in the transfer gel that had the characteristics of both solid and liquid medium rather than had the characteristics of liquid medium with high fluidity. The separated female & male gametophytes were homogenized and cultivated in the liquid medium where the best growth and maturation were obtain in 5 stages of water temperature (5, 10, 15, 20 and 25℃) and 4 stages of intensity of radiation (20, 40, 60 and 80 μ㏖·m^(-2)·s^(-1)); consequently, the best results were obtained at 20℃ of water temperature and 20 μ㏖·m^(-2)·s^(-1) of radiationintensity in female gametophyte and at 10℃ of water temperature and 60 μ㏖·m^(-2)·s^(-1) of radiation intensity in male gametophyte. After separating the propagated female & male gametophytes into 10 populations, the fertilization was made repeatedly for 3 times in 5 stages of water temperature (5, 10, 15, 20 and 25℃) and 4 stages of intensity of radiation (20, 40, 60 and 80 μ㏖·m^(-2)·s^(-1)) on 96 well plate; as a result, the highest fertilization rate was shown at 15℃ of water temperature and 20 μ㏖·m^(-2)·s^(-1) of radiation intensity and it demonstrated the feasibility of mass production. After preserving the separated female & male gametophytes while exchanging the medium at intervals of a week in liquid medium, it has been found that the gametophytes were preserved while maintaining female & male gametophytes with average size of 300 ㎛ up to 10-month cultivation; however, most of the gametophytes were not germinated in solid medium and were in a state of zoospore so that it was possible to preserve the zoospores themselves as well as female & male gametophytes that were germinated and grown and to preserve the gametophytes and species. It would suggest that Kjellmaniella crassifolia in danger of extinction could be possible to be mass-produced by fertilizing cultivated female & male gametophytes if necessary any time during the year, and it is believed that it is possible to build a gene bank by preserving the species of Kjellmaniella crassifolia in East Sea.
more목차
Abstract = i
List of tables = v
List of figures = vi
Ⅰ. 서론 = 1
Ⅱ. 재료 및 방법 = 4
1. 시료의 채집 및 배지 = 4
가. 채집 및 전처리 = 4
나. 배지 = 6
2. 유주자의 발아율 및 배우체와 아포체의 생장 = 8
가. 유주자의 발아율 = 8
나. 배우체의 생장과 성숙 = 9
다. 아포체의 생장 = 10
3. 배우체 분리 및 번식 = 11
가. 배우체 분리 = 11
나. 배우체 번식 = 11
4. 배우체 수정 = 12
5. 배우체 보존 = 12
Ⅲ. 결 과 = 13
1. 유주자의 발아율 = 13
가. 액체배지 = 13
나. 고체배지 = 15
1) Transfer gel = 15
2) Purified agar = 17
3) Agar = 19
4) High gel = 21
2. 배우체의 생장과 성숙 = 23
가. 액체배지 = 23
나. 고체배지 = 25
1) Transfer gel = 25
2) Purified agar = 27
3) Agar = 29
4) High gel = 31
3. 아포체의 생장 = 33
가. 액체배지 = 33
나. 고체배지 = 33
4. 배우체 분리 = 35
가. 액체배지 = 35
나. 고체배지 = 35
1) Transfer gel = 35
2) Purified agar = 36
3) Agar = 36
4) High gel = 36
5. 배우체 번식 = 38
6. 배우체 수정 = 42
가. 액체배지 = 42
나. 고체배지 = 43
7. 배우체 보존 = 45
Ⅳ. 고찰 = 46
Ⅴ. 요약 = 49
Ⅵ. 감사의 글 = 53
Ⅶ. 참고문헌 = 54
