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60Co 감마선 조사에 따른 속성장 및 환경내성에 강한 둥근전복과 오분자기의 신품종 개발에 관한 연구

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Abstract
Climate change by global warming has raised the surface temperature of
South Korea up by 1.03oC for the last 40 years, which is the fastest
rate in the world. Furthermore, the rise of temperature in the sea is
also found more obviously than ever. The rise causes the lead to
subtropical climate. As evidence, it has been observed that butter fish
(finny tribe in the Pacific) was caught in the net on Korean Peninsula.
More fishes that have origin in subtropical climate regions and
turbulence kind are captured. Furthermore, mitra squid Loligo chinesis,
which used to be caught during July and August, is now caught on
September. Judging from these, considerable changes are taking place in
marine ecology. The rise of water temperature works as a physical and
chemical stress factor to marine life. Particularly on abalone which is
one kind of Gastropoda, the rise has a great impact because the species
is limited to mobility and thus has very weak resistance to
environmental change.
Therefore, National Fisheries Research and Development Institute
(NFRDI) crossbred male Haliotis discus hannai with female Haliotis
sieboldii in 2004, both of which grow fast and are firmly resistant to
environmental change, in an attempt to develop an ew species that is
resistant to sudden change in environment. The developed species was sea
Haliotis discus hannai. Sea Haliotis discus hannai grows 20% faster thanHaliotis discus hannai and as fast as Haliotis sieboldii. In addition,
it was found that sea Haliotis discus hannai has higher level of
resistance to low water temperature(7℃) and high temperature (28℃)
than Haliotis discus hannai in water temperature resistance test.
For breeding methods, there are hybrid breeding, selection breeding and
mutation breeding. NFRDI used hybrid breeding to develop an abalone that
grows fast and has high resistance.
Mutation breeding processes organism with radiation and chemicals
because mutation takes place very rarely in organism. And mutants are
selected from their posterior filial generation. When radiation is
applied to it, the method is called radiation breeding.
When exposed to radiation, organism is mutated by the damage to part of
its chromosome and DNA, so that its information signaling system is
confused. Mutants are generated from the cells in which such change
brings about. Radiation breeding method is a very excellent technique in
developing new breeds and creating useful genetic resources. The method
has steadily produced good outputs for about 70 years since 1930s.
Currently about 2,600 mutated species are registered in FAO-LAEA
database (Kim, 2008). After the Convention on Biological Diversity,
securing biological genetic resources was considered to be the base of
national competitiveness. In this respect, we can say that it is urgent
to secure source technologies related to new agriculture biotechnologythat uses radiation and biotechnology.
The merits of radiation breeding include advantage of developing one or
two genetic traits, excellence in improving inferior traits of a
superior breed, more yields, better quality and higher resistance to
harmful insects than those of existing breeds. Unlike GMO that
artificially incorporates foreign genes into existing breeds, radiation
breeding has been verified for its safety. It has been widely used in
developing crops such as rice and bean, flowers like rose and
chrysanthemum and fruit trees. In addition, radiation breeding requires
less developing cost than other breeding technologies.
Furthermore, radiation breeding can be applied to plant breeds, to
where hybrid breeding can't be applied or for which GMO breeding method
hasn't been established. Therefore radiation breeding can be expected to
develop native plants and foreign plant breeds that South Korea lacks,
contributing to expanding the resource of breeds (genetic resources).
(Kang, 2007)
To determine the impact of low-dose irradiation with gamma ray on
Haliotis discus discus, the growth was observed through the
physiological change and breeding. The death of parent abalones by
irradiation with gamma ray has not occurred in all the experimental
plots from the domestication period to the egg collection period.
However, it was observed that the activity decreased in some of theparent shellfishes of the 20Gy and 25Gy experimental plots. It seems
that this phenomenon was due to the damage of DNA caused by irradiation
with gamma ray. Also, it is estimated that this phenomenon was caused
because it would consume energy to restore the damage of DNA.
As for those fertilized eggs that had been obtained by the artificial
egg collection, the presence of mutation of fertilized eggs were
observed under the microscope every hour after transferring the seawater
of 1L that contained the fertilized eggs for each experimental plot. The
mutation of fertilized eggs has not occurred at the control plot, 10, 15
and 20Gy; however, it has occurred at the 25Gy experimental plot. In the
developmental process of fertilized eggs, the form of mutated eggs was
observed at the 15, 20 and 25Gy with the exception of the control plot
and 10Gy, whereas the occurrence has not progressed further.
The extent of occurrence delay could be related to the spawning of nonfertilized
eggs in accordance with gamma ray amount and the sex maturity
ratings of abalone for irradiation with gamma ray. Also, this experiment
could verify the increase in the probability of mutation for the
fertilized eggs by the irradiation; however, it could not verify that
the morphological mutation of fertilized eggs was related to the
mutation of Juvenile abalone.
To measure the hatching rate and attachment rate, the fertilized eggs
were measured as they were housed and bred in the water tank that had been set for each experimental plot. In the case of hatching rate, it
was found to be approximately 50 percent at the control plot and the
experimental plots of 10 and 15Gy; however, it was found to be
approximately 30 percent at the experimental plots of 20 and 25Gy dose.
This study produced F1 Juvenile abalone by irradiating abalone with
gamma ray for the first time in South Korea and even applied for the
relevant patent.
Also, the analysis on the remaining amount in Juvenile abalone that had
been produced after being irradiated with gamma ray was performed. As
for the production of genes, the oligonucleotide primer as to Myostatin,
Caspase-8, HSP-70 mRNA that had been registered at NCBI in relation with
abalone were produced and synthesized, and then the expression level and
so on were investigated. As a result, no remaining amount of gamma ray
was detected in all the experimental plots of 10Gy, 15Gy and 20Gy, which
had been irradiated with gamma ray. It is believed that there would be
no residual substance because gamma ray was a light rather than a
substance residing in the body.
The expression of Caspase-8 mRNA that was immune gene expression in
accordance with irradiation with gamma ray was found in all the
experimental plots. Also, it was possible to confirm that the expression
level was higher in the 20Gy experimental plot than the other
experimental plots. It is believed from these results that theexperimental plot that had been irradiated with gamma ray 20Gy would
have a better environmental tolerance.
The expression of Myostatin mRNA that was growth gene was not found in
the control experimental plot; however, it could be confirmed that it
was expressed at a high level in the gamma ray 20Gy experimental plot.
It is believed from these results that a rapid growth of abalone
development would be possible by irradiation with gamma ray of 20Gy.
Moreover, it could be confirmed that the expression of heat shock
protein (HSP 70) mRNA, which was a stress protein, was found in the 20Gy
experimental plot for water temperature and the 20Gy experimental plot
for salinity. It is believed that the experimental plot that had been
irradiated with gamma ray of 20Gy had a better environmental tolerance
at 30℃ as for the stress of water temperature, whereas the experimental
plot that had been irradiated with gamma ray of 20Gy at 25psu as for the
stress of salinity had a better self-immune mechanism ability.
At last, it was confirmed that marine organisms had a good result by
using gamma ray as terrestrial organisms. Thus, it would be required to
conduct a study in breeding and immune function for other more breeds.
It should be necessary to conduct more diverse experimental analysis and
long-term studies for comprehensive review on various changes.
Author(s)
좌민석
Issued Date
2014
Awarded Date
2015. 2
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000007050
Alternative Author(s)
Min-Seok Jwa
Department
대학원 해양생명과학과
Table Of Contents
Ⅰ. 종합서론 1
Ⅱ. 60Co 감마선 조사가 둥근전복과 오분자기에 미치는 영향 6
1. 서 론 6
2. 재료 및 방법 9
1) 모패관리 9
1-1) 둥근전복 9
1-2) 오분자기 9
2) 감마선 조사용 케이스 제작 10
3) 감마선 조사 11
4) 혈림프(Hemolymph) 수 측정 12
5) 항산화 효소 측정 13
6) 통계처리 16
3. 결과 17
1) 감마선 선량에 따른 둥근전복 Hormesis 변화 17
1-1) 생존율 변화 17
1-2) 혈림프수 변화 18
1-3) 항산화효소 변화 19
2) 감마선 조사에 따른 오분자기 Hormesis 변화 21
2-1) 생존율 변화 21
2-2) 혈림프수 변화 22
2-3) 항산화효소 변화 23
4. 고 찰 25
Ⅲ. 60Co 감마선 조사 후 생산된 전복류 치패의 형질 변화 29
1. 서론 29
2. 재료 및 방법 31
1) 실험재료 및 사육관리 31
1-1) 둥근전복 31
1-2) 오분자기 31
2) 부착규조류 배양 32
3) 감마선 조사 후 종자생산 33
4) 난발생 과정 조사 34
5) 수정률 및 부화율 측정 34
6) 유생관리 34
7) 부착률 측정 35
8) 성장률 측정 35
9) 성장 유전자 mRNA 발현 분석 36
10) RT-PCR 37
11) Thermoluminescence(TL) 측정 38
12) 통계처리 39
3. 결과 40
1) 사육관리 40
1-1) 둥근전복 40
1-2) 오분자기 40
2) 먹이생물의 공급 41
3) 난발생 과정 변화 42
4) 수정란 수 및 수정률 변화 45
4-1) 둥근전복 45
4-2) 오분자기 47
5) 비정상 발생난의 계수 49
5-1) 둥근전복 49
5-2) 오분자기 50
6) 부화율 및 부착률 변화 51
6-1) 둥근전복 51
6-2) 오분자기 53
7) 성장률 변화 55
7-1) 둥근전복 55
7-2) 오분자기 56
8) 성장유전자 MYOSTATIN mRNA 변화 57
9) Thermoluminescence(TL) 변화 58
4. 고찰 59
Ⅳ. 감마선 조사 후 생산된 전복류 F1 치패의 환경내성 변화 연구 63
1. 서론 63
2. 재료 및 방법 67
1) 수온, 염분 내성측정 67
2) 생존율 측정 67
3) 항산화효소 측정 68
4) Respiratory burst activity 측정 69
5) Phenoloxidase activity 측정 69
6) Lysozyme activity 측정 70
7) 면역유전자 mRNA 70
8) RT-PCR 분석 71
3. 결과 73
1) 둥근전복 고수온 내성 변화 73
1-1) 생존율 73
1-2) 항산화효소 변화 74
1-3) Respiratory burst activity 변화 76
1-4) Phenoloxidase activity 변화 77
1-5) Lysozyme activity 변화 78
2) 오분자기 고수온 내성 변화 79
2-1) 생존율 79
2-2) 항산화효소 변화 80
2-3) Respiratory burst activity 변화 82
2-4) Phenoloxidase activity 변화 83
2-5) Lysozyme activity 변화 84
3) 둥근전복 저염분 내성 변화 85
3-1) 생존율 85
3-2) 항산화효소 변화 86
3-3) Respiratory burst activity 변화 88
3-4) Phenoloxidase activity 변화 89
3-5) Lysozyme activity 변화 90
4) 오분자기 저염분 내성 변화 91
4-1) 생존율 91
4-2) 항산화효소 변화 92
4-3) Respiratory burst activity 변화 94
4-4) Phenoloxidase activity 변화 95
4-5) Lysozyme activity 변화 96
5) 면역유전자 CASPASE-8 mRNA 변화 97
6) 열충격 단백질(HSP 70) mRNA 변화 98
4. 고찰 100
Ⅴ. 종합고찰 104
Ⅵ. 요약 109
Ⅶ. 참고 문헌 111
Degree
Doctor
Publisher
제주대학교 대학원
Citation
좌민석. (2014). 60Co 감마선 조사에 따른 속성장 및 환경내성에 강한 둥근전복과 오분자기의 신품종 개발에 관한 연구
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