[보고서]친환경 LED 광원을 이용한 고품질 어류의 생산기술 개발

친환경 LED 광원을 이용한 고품질 어류의 생산기술 개발
Development of the high valued product of marine fish using an environmental-friendly LED light 원문보기

보고서 정보
주관연구기관	한국해양대학교 Korea Maritime University
보고서유형	최종보고서
발행국가	대한민국
언어	한국어
발행년월	2013-06
과제시작연도	2011
주관부처	해양수산부 Ministry of Oceans and Fisheries
등록번호	TRKO201400000109
과제고유번호	1545002870
사업명	수산실용화기술개발
DB 구축일자	2014-05-07

초록 ▼

○ 연구 결과
1.LED 광원의 제작 및 해수 관상 어종의 선정/광반응 특성 파악
- 빛의 파장과 강도를 조절할 수 있는 사육 시스템 설치용 LED 광원 모듈의 제작
- 고부가가치 해수 관상어인 2종의 흰동가리류와 파랑돔 및 금붕어를 대상 어종으로 선정하여 연구를 개시 (부가적으로 넙치를 대상으로 한 실험도 실시)
- 일주기에 따른 혈중 melatonin 농도 변화 및 타입별 melatonin 수용체의 발현 변화 조사
- 광반응 관련 유전자(흰동가리: 8종류, 파랑돔: 4종류, 넙치: 11종류)의 클로닝
2.LED 광원별 해수 관상어종의 생체리듬 분석 및 파장별 광반응 특성 조사
- 흰동가리/파랑돔 망막에서 특이적으로 발현하는 Rhodopsin(RH)를 확인
- 대조구(형광등)및 LED 광원(Green, Blue 및 Red)실험구별,흰동가리의 광반응 유전자인 melatonin수용체 1(MT1)mRNA의 발현량 변화를 통한 생체리듬 파악
- LED Blue/Green파장이 흰동가리의 절식에 따른 스트레스 감소에 효과적임을 입증
- LED 광원별 흰동가리 및 파랑돔의 광반응 및 생체리듬 분석을 위해 광수용체인 rhodopsin과 coneopsin,생체리듬 유전자인 Period2와 Cryptochrome1의 분석을 통해 빛에 의하여 시계유전자가 생체리듬을 조절하고 있음을 확인
3. LED 광원별 해수 관상어종 및 강도다리의 성장,성숙,스트레스 및 체색 변화 조사
- LED 광원별로 6개월과 4개월간 흰동가리와 파랑돔을 사육한 결과,0.3w/m2의 세기(강도)의 Blue및 Green파장에서 빠른 성장과 성 성숙 촉진 효과를 확인
- 실험실 수준이 아닌 참여기업인 화남수산의 양식 현장에 실질적으로 적용하여, Blue LED 광원으로 6개월간 강도다리를 사육한 결과, 형광등을 설치한 일반 강도다리에 비하여 전장과 체중 모두 약 30% 정도의 빠른 성장이 관찰.또한,총 6개월간의 사육 실험임에도 불구하고, Blue LED 광원으로 사육한 강도다리가 약 2개월 정도 빠른 성장을 보인 점으로 보아, Blue 파장에서의 빠른 성장 효과를 실제의 양식 현장에서도 확인
- Blue 및 Green LED 파장이 12일간의 절식에 의해 흰동가리 체내에서 생성된 산화 스트레스 감소 효과:항산화 효소(SOD,CAT 및 GPX)의 발현/활성,활성산소(H2O2)농도 및 근육 내의 지질 과산화 (LPO)농도의 변화를 통하여 산화 스트레스 감소 효과를 확인
- LED Red파장에서 흰동가리의 체내에서 활성산소(H2O2)와 지질과산화(LPO)수치가 높아지는 것으로 보아,Red파장은 흰동가리의 산화 스트레스를 증가시키는 것으로 확인. 즉, Red 파장은 어류 양식 환경으로 적합하지 않는 파장임을 확인
- 비록 해수 관상어종은 아니지만, LED 광원별로 금붕어의 뇌와 피부세포 등을 대상으로 체색변화를 조사한 결과, Blue/Green파장의 어류 체색 조절(변화)효과를 확인

Abstract ▼

Ⅳ. Results
1. LED lights production and selection of ornamental fish/light reaction understanding
-Production of LED modules as raise system with regulated wavelengths and intensities
-Selection of high-value ornamental fish to yellowtail clownfish and blue damselfish as experimental fish
-Investigation of plasma melatonin and melatonin receptor types mRNA expression changes by diurnal rhythm
2. Investigation of circadian rhythm analysis and light reaction of fish using LED lights
-Cloning of genes with light reaction (yellowtail clownfish: 8, blue damsel: 4, olive flounder: 11)
-MT1 mRNA was highly expressed in Green/Blue LED groups of the yellowtail clownfish, which suggests that short wavelengths influence the maintenance of circadian rhythms
-Investigation of photoreceptors, rhodopsin and cone opsin, and circadian rhythm genes, Period2 and Cryptochrome1 showed that the expressions were changed in yellowtail clownfish and blue damsel by LED lights
3.Understandings the effect of ornamental fish growth, sex maturation, stress and body color using LED lights
-Examination showed that the Green/Blue LED lights enhanced the fish growth and sex maturation
-Examination showed that the Red LED light induces oxidative stress and the light is inconsistence to fish aquaculture environment
-Blue/Green LED lights regulate body color, and affect the endocrine changes of goldfish

목차 Contents

표지 ... 1
제 출 문 ... 2
요 약 문 ... 3
SUMMARY ... 4
CONTENTS ... 5
목 차 ... 6
제 1장 연구개발과제의 개요 ... 7
제 1절.연구개발의 필요성 ... 7
제 2절.연구개발 목표 및 내용 ... 8
제 2장 국내외 기술개발 현황 ... 9
제 3장 연구개발수행 내용 및 결과 ... 10
I. 사육 시스템용 LED 양식조명의 제작 및 보완 ... 10
II. LED 광원별 melatonin 수용체의 발현 비교 분석 ... 11
III. LED 광원별 산화 스트레스 비교 분석 ... 14
IV. 생체 리듬의 비교 및 분석 ... 16
V. LED 광원별 스트레스 및 생체리듬의 변화 비교 및 분석 ... 22
VI. LED 광원별 및 세기별 어류의 성장 변화 비교 ... 33
VII. LED 광원별 및 세기별 어류의 성숙 변화 비교 및 분석 ... 40
VIII. LED 광원별 어류의 체색 변화 비교 ... 53
IX. LED 광원별 어류의 스트레스 변화 비교 ... 59
X. LED 광원별 실험어의 안구 적출에 따른 생체리듬의 변화 및 망막의 광수용체 유전자의 발현 비교 ... 70
XI. BlueLED 조명등 설치 양어장에서 강도다리의 성장 차이 조사 ... 81
제 4장 목표달성도 및 관련분야에의 기여도 ... 83
제 1절 목표달성도 ... 83
제 2절 관련분야에의 기여도 ... 84
제 5장 연구개발 성과 및 성과활용 계획 ... 85
제 1절 연구개발 성과 ... 85
제 2절 연구결과의 활용계획 ... 88
제 6장 연구개발과정에서 수집한 해외과학기술정보 ... 90
제 7장 연구시설‧장비 현황 ... 91
제 8장 참고문헌 ... 92
[별첨 1] 연구개발보고서 초록 ... 96
끝페이지 ... 102

표/그림 (59)

표 Spectral profiles of blue, green and red LEDs used in this experiment.
표 Different colors of LED light for indoor aquaculture tank.
표 Photograph showing the rearing of fish under different colors of LED light; blue,green,andred,respectively.
표 Primers for quantitative real-time PCR.
표 Changes in the expression levels ofMT1 mRNA in the brain of yellowtail clownfish under lighting conditions using red
표 Primers for quantitative real-time PCR
표 Changesin the expression levels of SOD and CAT mRNA in thelive
표 Primers used for quantitative real-time PCR of blue damsel
표 Tissue distribution of rhodopsin(RH) mRNAs in yelowtail clownfish (CK) and blue damsel(BD). β-actin mRNA was amplified to verify the integrity of each samples.
표 Diurnal variation in the level of three subtypes of MT mRNAs, as measured by quantitative real-time PCR, in the brain of yellowtail clownfish and blue damsel.
표 ELISA melatonin measurement in yellowtail clownfish and blue damsel plasma during the daily light-dark cycle by microplate reader.
표 Changesin the expression levels of Per2, Cry1 mRNA in the yelowtail clownfish and blue damsel brain as measured by quantitative real-time PCR.
표 Primers sued for quantitative real-time PCR of yellowtail clownfish.
표 Primers used for quantitative real-time PCR of blue damsel.
표 Changes in the H2O2 and LPO levelsin plasma and muscle
표 Changes in the CAT mRNA expression levels in liver of blue damsel under red (R),green (G),blue (B),and purple (P)LEDs,a simulated natural photoperiod (SNP),and three intensities
표 Changes in the SOD and CAT activity levels in liver of blue damsel under red (R),green (G),blue (B),and purple (P)LEDs,a simulated natural photoperiod (SNP),and three intensities
표 Enzyme-linked immunosorbentassay melatonin plasma levels in yellowtail clownfishandbluedamselunderlightingconditionsusing red(R),green(G),blue(B) LEDs and a simulated naturalphotoperiod (SNP),as measured by microplate reader
표 Changes in the H2O2 and LPO levels in plasma and muscle of yellowtail clownfish under red(R),green(G),blue(B),andpurple(P)LEDs,a simulated natural photoperiod (SNP), and three intensities.
표 Changes in the total length of yellowtail clownfish (A.clarki,YC) and blue damsel(C.parasema,BD)were reared under fluorescence bulb,red,blue,andgreen LED lights.
표 Gonadosomatic index for blue damsel under red (R),green (G),blue(B),and purple (P) LEDs, a simulated natural photoperiod(SNP), and three intensities.
표 Comparison of the GH amino acid sequence of yellowtail clownfish
표 Changes in the total length of yellowtail clownfish reared undera simulated natural photoperiod and red,blue,and green light-emitting diode lights
표 Changes in the total body length of yellowtail damselfish ,which were reared for 4 months under a simulated natural photoperiod (SNP),as well as red (R),green (G),andblue(B)LED lights.
표 Changes in the expression levels ofGH mRNA in the pituitary before(A) and after injection with melatonin (B)underlighting conditions using red (R),green (G),and blue(B)LEDs and a simulated natural photoperiod (SNP),as measured by quantitative real-time PCR.
표 Primers sued for quantitative real-time PCR of yellowtail damselfish
표 VTG and ERα mRNA expression levels in the livers and gonads of yellowtail damselfish under lighting conditions using a simulated natural photoperiod (SNP),red (R),green (G),blue (B),purple (P)LED lights,and low (L),middle (M),high (H) intensities.
표 Changes in gonadosomatic index (GSI) of the yellowtail damselfish under lighting conditions using a simulated natural photoperiod (SNP),red (R),green (G),blue(B),purple(P)LED lights,andlow (L),middle(M),high (H)intensities
표 Changes in gonad histology of the yellowtail damselfish (Chrysiptera parasema)under lighting conditions using a simulated natural photoperiod (SNP)(A),red(B),green(C),blue(D),purple(E)LED lights.
표 Plasma E2 hormone levels of the yellowtail damselfish under lighting conditions using a simulated natural photoperiod (SNP),red (R),green (G),blue(B), purple (P)LED lights, and low (L),middle (M),high (H)intensities.
표 Changes in the expression levels of sGnRH (A)andcGnRH-II (C)mRNA in the hypothalamus, and sGnRH (B)and cGnRH-II(D)in the brain cells in goldfish under lighting conditions using red (R),green (G)LEDs and white fluorescent bulb (Cont.),as measured by quantitative real-time PCR.
표 Changesin the expression levels of GTHα protein and mRNA (A),GTHβ1 and plasma FSH (B),GTHβ2 and plasma LH (C) levels in the pituitary and plasma of goldfish under lighting conditions using red(R),green(G) LEDs and white fluorescent bulb(Cont.),as measured by quantitative real-time PCR and microplate reader.
표 Changes in the expression levels of GTHα,GTHβ1,and GTHβ2 mRNA expression levels in the brain cells of goldfish under lighting conditions using red(R),green (G) LEDs and white fluorescent bulb (Cont.),as measured by quantitative real-time PCR
표 Changes in the expression levels ofKiss1 mRNA in the hypothalamus (A) and the levels in the brain cells (B)mRNA expression levels in the goldfish under lighting conditionsusing red (R),green (G)LEDsand whitefluorescentbulb (Cont.), as measured by quantitative real-time PCR.
표 Changes in gonad histology of goldfish under different lighting conditions using a white fluorescentbulb (Cont.)(A),as wellas red (B),and green (C)LED lights. PNS:Peri-nucleolusstage,OYS:Oocytesintheyolkstage,Scalebar=20um
표 Changes in the levels of plasma 17α-hydroxypregnenolone in goldfish under lighting conditionsusing red (R),green (G)LEDs and white fluorescent bulb (Cont.), as measured by microplate reader.
표 Primers used for quantitative real-time PCR of goldfish
표 Changesin the expression levels of Opn4m mRNA in the retina of goldfish under lighting conditions using red (R),green (G),purple (P) LEDs and white fluorescent bulb (Cont.), as measured by quantitative real-time PCR.
표 Changesin the expression levels of MCH mRNA in the pituitary of goldfish under lighting conditions using red (R),green (G),purple (P) LEDs and white fluorescent bulb (Cont.),as measured by quantitative real-time PCR.
표 Changes in the expression levels of POMC mRNA in the pituitary of goldfish under lighting conditions using red (R),green (G),purple (P) LEDs and white fluorescent bulb (Cont.),as measured by quantitative real-time PCR.
표 Primers used for quantitative real-time PCR of goldfish
표 Changesin the expression levels of SOD (A,D),CAT (B,E),and GPX (C, F) mRNA in the liver during thermal change [low-water temperature (A,B,C); high-watertemperature(D,E,F)]of goldfish under lighting conditions using red (R), green(G),blue(B),and purple mix(PM)light-emitting diodes and a simulated natural photoperiod (SNP),asmeasured by quantitativereal-timePCR.
표 Changes in the activity of SOD (A,D),CAT (B,E),andGPX (C,F)mRNA in the liver during thermalchange [low-water temperature (A,B,C);high-water temperature(D,E,F)]of goldfish under lighting conditions using red (R),green (G), blue (B), and purple mix (PM) light-emitting diodes and a simulated natural photoperiod (SNP), as measured by microplate reader.
표 Changesin thePRDX mRNA expression in theliverduring thermalchange [low-water temperature (A);high-water temperature (B)]ofgoldfish under lightin conditions using red (R),green (G),blue (B),and purple mix (PM)light-emitting diodes and a simulated natural photoperiod (SNP), as measured by quantitative real-timePCR
표 Changesin theHSP70mRNA expression in theliverduring thermalchange [low-water temperature (A);high-water temperature (B)]ofgoldfish under lightin conditions using red (R),green (G),blue (B),and purple mix (PM)light-emittin diodes and a simulated natural photoperiod (SNP), as measured by quantitativ real-timePCR
표 Plasma H2O2 concentrations (A,C)and LPO levels (B,D)during thermal chang [low-water temperature (A,B);high-water temperature (C,D)]ofgoldfish under lighting conditions using red (R),green (G),blue (B),and purple mix (PM) light-emitting diodes and a simulated naturalphotoperiod,as measured by microplate reader
표 Plasma melatonin levels during thermal change (low-water temperature-A high-water temperature-B)of goldfish under lighting conditions using red (R),green (G),blue (B),and purple mix (PM) light-emitting diodes and a simulated natural photoperiod,as measured by microplate reader.
표 Primers used for quantitative real-time PCR of yellowtail clownfish
표 Primers used for quantitative real-time PCR of yellowtail damselfish
표 Primers used for quantitative real-time PCR of yellowtail clownfish
표 Changes in the Per2 and Cry1 expression levels in brain of yellowtail clownfish and eye-removed yellowtail clownfish under red(R),green(G),andblue(B) LEDs, a simulated natural photoperiod (SNP),and three intensities.
표 Enzyme-linked immunosorbent assay melatonin plasma levels in yellowtail clownfish and eye-removed yellowtail clownfish under lighting conditions usingre red(R), green(G),blue(B)LEDs and a simulated natural photoperiod (SNP),as measured by microplate reader.
표 Changesin the red and green opsin expression levels in retina of yellowtail damselfish under red (R), green (G), and blue (B) LEDs, a simulated natural photoperiod(SNP),and three intensities.
표 Changes in the expression levels of RH mRNA and protein in the retina of intact fish (a) and the ophthalmectomized fish (b)mRNA in the retina under lighting conditions using red (R),green (G),blue (B)LEDs and and white fluorescent bulb (Cont.).
표 Immunohistochemical localization of retina RH-immunoreactivity (RH-IR)in cross sections of intact and ophthalmectomized fish under lighting conditionsusing red (R),green(G),blue(B)LEDs and and white fluorescent bulb(Cont.)at ZT8 of second day
표 Changes in the expression levels of MT1 mRNA and protein in the brain of the intact fish (a) and the ophthalmectomized fish (b) mRNA in the brain of yellowtail clownfish under lighting conditions using red (R),green (G),blue(B)LEDsand and white fluorescent bulb (Cont.), as measured by quantitative real-time PCR.
표 Enzyme-linked immunosorbent assay melatonin plasma levels in the intact fish (a) and levels in the ophthalmectomized fish (b) in yellowtail clownfish under lighting conditions using red (R),green (G),blue (B)LEDs and and white fluorescent bulb (Cont.), as measured by microplate reader.
표 Changes in the total length and weight of starry flounder,which were reared under a white fluorescent bulb and blue LED lights
표 Photograph showing the rearing system of fish under different colors of LED light inTakemura's Lab.

과제명(ProjectTitle) :	-
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연합인증

친환경 LED 광원을 이용한 고품질 어류의 생산기술 개발
Development of the high valued product of marine fish using an environmental-friendly LED light 원문보기