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제 1 장연구개발과제의개요

1 (2002 2003) 2 (2003 2004) 3 (2004 2005)

제 2 장국내외기술개발현황

제 3 장종보존을위한동결및해동조건의최적화

μ μ μ

Survival rate (%) 100 80 60 40 20 0 50%S.W.+DMSO 100%S.W.+DMSO 0 5 10 15 DMSO concentration (%) Survival rate (%) 100 80 60 40 20 0 S.W. S.W.+5%DMSO 0 25 50 75 100 Seawater concentration (%)

100 80 sorbitol sucrose s orbitol+5%dmso s ucros e+5%dmso Survival rate (%) 60 40 20 0 0 0.1 0.5 1 Concentration (M)

100 80 Survival(%) 60 40 20 0 Survival(%) 100 80 60 40 2.50% 5% 10% 15% 20 0-20 -40-60 -80 Temperature ( )

100 80-40 -40 LN Survival(%) 60 40 20 0 0 2.5 5 10 15 20 DMSO Concentration(%)

100 80 Suvival(%) 60 40 20 0 100 80 0.5 /min 1 /min 2 /min 4 /min Survival(%) 60 40 20 0-20 -40-60 -80 Temperature( )

100 80 Survival(%) 60 40 20 0-40 -40 LN 0 0.5 1 2 4 Cooling rate( /min)

100 80 Survival(%) 60 40 20 0 100 80 Survival(%) 60 40 20 0 2 /min 1 /min 0.5 /min 0 10 20 40 60 TIME(min)

100 80 Survival(%) 60 40 20 0-40 -40 LN 0 60 120 240 Time(min)

100 80 Survival rate(%) 60 40 20 0 0 100 200 300 400 500 Time(day s)

35 30 prefreezing prefreezing- LN SURVIVAL(%) 25 20 15 10 5 0-20 -30-40 -50-60 PREFREEZING TEMPERATURE( )

40 35 prefreezing prefreezing- LN 30 SURVIVAL(%) 25 20 15 10 5 0-20 -30-40 -50-60 PREFREEZING TEMPERATURE( )

100 prefreezing prefreezing- LN 80 SURVIVAL(%) 60 40 20 0-20 -30-40 -50-60 PREFREEZING TEMPERATURE( )

80 prefreezing prefreezing- LN 60 SURVIVAL(%) 40 20 0-20 -30-40 -50-60 PREFREEZING TEMPERATURE( )

제 4 장원종묘와동결보존종묘의비교생물학적실험

Temperature( ) 30 25 20 15 10 5 0 12 10 DO( mg /l) 8 6 4 2 0 38 36 Salinity( ) 34 32 30 28 Nov Dec. Jan. Feb. Mar. Apr. May Month

Blade length (Cm) 20 18 16 14 12 10 8 6 4 2 pre-cryopreservations post-cryopreservations Nov. Dec. Jan. Feb. Mar. Month 12 10 Blade width (Cm) 8 6 4 2 0 pre-cryopreservations post-cryopreservations Nov. Dec. Jan. Feb. Mar. Month

350 Blade length (Cm) 300 250 200 150 100 pre-cryopreservations post-cryopreservations Feb. Mar. Apr. May Jun. Month 25 Blade width (Cm) 20 15 10 5 0 pre-cryopreservations post-cryopreservations Feb. Mar. Apr. May Jun. Month

18 16 Blade length(cm) 14 12 10 8 6 4 2 0 pre-cryopreservations post-cryopreservations Nov. Dec. Jan. Feb. Mar. Month Blade width(cm) 50 45 40 35 30 25 20 15 10 5 0 pre-cryopreservations post-cryopreservations Nov. Dec. Jan. Feb. Mar. Month 25 Upper stipe length (Cm) 20 15 10 5 0 pre-cryopreservations post-cryopreservations Nov. Dec. Jan. Feb. Mar. Month

55 Total length (mm). 50 45 40 35 30 25 20 pre-cryopreservations post-cryopreservations Mar. Apr. May Jun. July Month 30 25 Branch number 20 15 10 5 0 pre-cryopreservations post-cryopreservations Mar. Apr. May Jun. July Month

55 50 Total length (mm). 45 40 35 30 25 20 pre-cryopreservations post-cryopreservations Mar. Apr. May Jun. July Month 40 35 Branch number 30 25 20 15 10 5 0 pre-cryopreservations post-cryopreservations Mar. Apr. May Jun. July Month

30 Blade length (Cm) 25 20 15 10 5 0 pre-cryopreservations post-cryopreservations Nov. Dec. Jan. Feb. Mar. Month 2.5 Blade width (mm) 2 1.5 1 0.5 pre-cryopreservations post-cryopreservations Nov. Dec. Jan. Feb. Mar. Month

Total blade length (mm). 160 140 120 100 80 60 40 20 0 pre-cryopreservations post-cryopreservations Mar. Apr. May Jun. July Month Branch number 90 80 70 60 50 40 30 20 10 0 pre-cryopreservations post-cryopreservations Mar. Apr. May Jun. July Month

λ λ

100 80 pre-cry opres ervations pos t-cry opres ervations Unit(g/100g) 60 40 20 0 Mois ture protein Lipid As h Compos ition Unit(mg/100g). 250 200 150 100 50 0 pre-cryopreservations post-cryopreservations Taur Asp Thr Ser Asn Glu Sarc Pro Gly Ala aaba Val Cys Met cysth Ile Leu Tyr b-ala Phe Amm Hylys Lys Arg Amino Acids Unit(mg/100g). 3000 2500 2000 1500 1000 500 0 Asp Thr Ser Glu pro Gly Ala Met Ile Leu Tyr Phe His Lys Amm Arg Amino Acids pre-cryopreservations post-cryopreservations

Unit(g/100g) 100 80 60 40 20 0 Moisture protein Crude fat Ash Composition pre-cryopreservations post-cryopreservations Unit(mg/100g). 250 200 150 100 50 0 pre-cryopreservations post-cryopreservations Taur Asp Thr Ser Asn Glu Sarc Pro Gly Ala aaba Val Cys Met cysth Ile Leu Tyr b-ala Phe Amm Hylys Lys Amino Acids Unit(mg/100g). 3500 3000 2500 2000 1500 1000 500 0 Asp Thr Ser Glu pro Gly Ala Val Met Ile Leu Tyr Phe His Lys Amm Arg Amino Acids pre-cryopreservations post-cryopreservations

Unit(g/100g) 100 80 60 40 20 0 Moisture protein Crude fat Ash Composition pre-cryopreservations post-cryopreservations Unit(mg/100g). 250 200 150 100 50 0 Taur Asp Thr Ser Asn Glu Sarc Pro Gly Ala aaba Val Cys Met cysth Ile Leu Tyr b-ala Phe Amm Hylys Orn Lys Amino Acids pre-cryopreservations post-cryopreservations 3000 2500 pre-cry opres ervations pos t-cry opres ervations Unit(mg/100g). 2000 1500 1000 500 0 Asp Thr Ser Glu pro Gly Ala Met Ile Leu Tyr Phe His Lys Amm Arg Amino Acids

Unit(g/100g) 100 80 60 40 20 0 Moisture protein Crude fat Ash Composition pre-cryopreservations post-cryopreservations Unit(mg/100g). 250 200 150 100 50 0 pre-cryopreservations post-cryopreservations Taur Asp Thr Ser Asn Glu Sarc Pro Gly Ala aaba Val Cys Met cysth Ile Leu Tyr b-ala Phe Amm Hylys Lys Amino Acids Unit(mg/100g). 3000 2500 2000 1500 1000 500 0 Asp Thr Ser Glu pro Gly Ala Val Met Ile Leu Tyr Phe His Lys Amm Arg Amino Acids pre-cryopreservations post-cryopreservations

Uint(g/100g) 100 80 60 40 20 0 Moisture protein Crude fat Ash Composition pre-cryopreservations post-cryopreservations Unit(mg/100g). 300 250 200 150 100 50 0 Taur Asp Hypro Thr Ser Asn Glu Sarc Pro Ala Val Met cysth Ile Leu Tyr b-ala Phe gaba Ethan Amm Amino Acids pre-cryopreservations post-cryopreservations Unit(mg/100g). 3000 2500 2000 1500 1000 500 0 Asp Thr Ser Glu pro Gly Ala Val Met Ile Leu Tyr Phe His Lys Amm Arg Amino Acids pre-cryopreservations post-cryopreservations

100 80 pre-cry opres ervations pos t-cry opres ervations Unit(g/100g) 60 40 20 0 Mois ture Protein Lipid As h Compos ition Unit(mg/100g). 300 250 200 150 100 50 pre-cry opres ervations pos t-cry opres ervations 0 Taur Asp Thr Ser Asn Glu Sarc Pro Gly Ala aaba Val Cys Met cysth Amino Acids Ile Leu Tyr b-ala Phe gaba Amm Hylys Lys Unit(mg/100g). 3000 2500 2000 1500 1000 500 pre-cryopreservations post-cryopreservations 0 Asp Thr Ser Glu pro Gly Ala Val Met Ile Leu Tyr Phe His Lys Amm Arg Amino Acids

100 80 pre-cry opres ervations pos t-cry opres ervations Unit(g/100g) 60 40 20 0 Mois ture Protein Lipid As h Compos ition Unit(mg/100g). 200 150 100 50 pre-cryopreservations post-cryopreservations 0 Taur Asp Hypro Thr Ser Asn Glu Sarc Pro Gly Ala Val Ile Leu Tyr b-ala Phe gaba Amm His Arg Amino Acids Unit(mg/100g). 2000 1500 1000 500 pre-cryopreservations post-cryopreservations 0 Asp Thr Ser Glu pro Gly Ala Val Met Ile Leu Tyr Phe His Lys Amm Arg Amino Acids

제 5 장해조류의종보존을위한동결및해동공정에서의 열특성연구

1) 마이크로슬라이스부 디지털신호 5) 신호발생기 (PC) 동기신호 2) 단면영상촬영장치 (CCD 카메라 ) 단면화상신호 3) 영상기억장치 (LD) ) 차원영상구축장치 (W/S) 영상 DATA 6) 모니터 관찰 관찰자

-10-15 Temperature ( ) -20-25 -30-35 On set piont Maximum point Cease point -40 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Cooling rate ( /min) -10-15 Temperature ( ) -20-25 -30-35 On set point Maximum point Cease point -40 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Cooling rate ( /min) -10-15 Temperature ( ) -20-25 -30-35 On set point Maximum point Cease point -40 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Cooling rate ( /min)

14 7.5 Temperature Difference ( ) 12 10 8 6 4 2 Temperature difference Freezing time 7.0 6.5 6.0 5.5 5.0 4.5 4.0 Freezing time (min) 0 3.5 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Cooling rate ( /min) 10 10 Temperature Difference ( ) 8 6 4 2 Temperature difference Freezing time 9 8 7 6 5 Freezing time (min) 0 4 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Cooling rate ( /min) 14 7.2 Temperature Difference ( ) 12 10 8 6 4 2 Temperature difference Freezing time 7.0 6.8 6.6 6.4 6.2 6.0 5.8 Freezing time (min) 0 5.6 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Cooling rate ( /min)

200 Freezing Latent heat, L(J/g) 100 153.58 146.67 123.08 0 U. pinnatifida P. seriata S.fulvellum

20000 Entrophy, S (J/mole K) 18000 16000 14000 12000 10000 8000 6000 4000 2000 Porphyra seriata Undaria pinnatifida Sargassum fulvellum 0 0.0 0.5 1.0 1.5 2.0 2.5 Cooling rate, V f ( /min) 5e+6 Entalphy,H (J/mole) 4e+6 3e+6 2e+6 1e+6 Porphyra seriata Undaria pinnatifida Sargassum fulvellum 0 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Cooling rate, V f ( /min) 13600 Free energy, F (J/mole) 13400 13200 F=60.424T+14402.048 Cooling rate 13000 0.1 0.2 12800 0.5 0.75 1.0 1.25 12600 1.5 1.75 2.0 Regression 12400-30 -28-26 -24-22 -20-18 -16-14 Temperature, T ( )

Free energy, F ( J/mole) 13500 13400 13300 13200 13100 13000 12900 12800 F=60.503T+14403.565 12700-28 -26-24 -22-20 -18-16 -14 Temperature, T ( ) Cooling rate 0.1 0.2 0.5 0.75 1.0 1.25 1.5 1.75 2.0 Regression Free energy, F (J/mole) 13500 13400 13300 13200 F=60.540T+14404.228 13100 Cooling rate 13000 0.1 0.5 1.0 12900 2.0 Regression 12800-28 -26-24 -22-20 -18-16 -14 Temperature, T ( ) 13600 Free energy, F (J/mole) 13400 13200 13000 12800 12600 F=60.424T+14402.048 12400-30 -28-26 -24-22 -20-18 -16-14 Temperature, T ( )

10 5 Temperature ( ) 0-5 -10-15 -20 On set point Maximum point Cease point -25-30 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Thawing rate ( /min) 10 5 Temperature ( ) 0-5 -10-15 -20 On set point Maximum point Cease point -25-30 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Thawing rate ( /min) 10 5 Temperature ( ) 0-5 -10-15 -20 On set point Maximum point Cease point -25-30 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Thawing rate ( /min)

32 45 Temperature Difference ( ) 30 28 26 24 22 20 18 16 14 Temperature difference Thawing time 40 35 30 25 20 15 Thawing Time (min) 12 10 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Thawing rate ( /min) 32 32 Temperature Difference( ) 30 28 26 24 22 20 18 16 Temperature difference Thawing time 30 28 26 24 22 20 18 16 14 Thawing Time (min) 14 12 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Thawing rate ( /min) 30 60 Temperature Difference ( ) 28 26 24 22 20 18 Temperature difference Thawing time 50 40 30 20 Thawing Time (min) 16 10 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 Thawing rate ( /min)

200 Thawing Latent heat, L(J/g) 100 128.37 118.53 94.42 0 U. pinnatifida P. seriata S. fulvellum

0-50000 Enthalpy, H(J/mole) -100000-150000 -200000-250000 Porphyra seriata Undaria pinnatifida Sargassum fulvellum -300000 0.0 0.5 1.0 1.5 2.0 2.5 Thawing rate, vt( /min) 0-200 Entropy, s(j/mole K) -400-600 -800-1000 Porphra seriata Undaria pinnatifida Sargassum fulvellum -1200 0.0 0.5 1.0 1.5 2.0 2.5 Thawing rate, vt( /min) 13900 13800 Free energy, F(J/mole) 13700 13600 13500 13400 13300 13200 Porphyra seriata Undaria pinnatifida Sargassum fulvellum 13100 0.0 0.5 1.0 1.5 2.0 2.5 Thawing rate, vt ( /min)

제 6 장목표달성도및관련분야에의기여도

제 7 장연구개발결과의활용계획

제 8 장참고문헌

Appendix

30 20 Temperature( ) 10 0-10 -20 0 25 50 75 92 97 Time(min) -30-40 -50