표제지
제출문
요약문
SUMMARY
CONTENTS
목차
제1장 연구개발과제의 개요 13
1. 기술적 필요성 13
2. 경제·산업적 필요성 14
3. 사회·문화적 필요성 16
4. 연구의 목적 및 범위 16
제2장 국내외 기술개발 현황 17
1. 수확후 선도유지를 위한 방안 17
2. 국내 연구 현황 18
3. 국외 연구 현황 19
4. 앞으로의 전망 19
제3장 연구개발수행 내용 및 결과 20
제1절 서론 20
제2절 연구수행방법 23
제3절 연구수행 내용 및 결과 37
I. 복숭아 병원성 진균 저해 활성 보유균주의 탐색 37
1. Monilinia fructicola, Rhizopus nigricans, Penicillium expansum, Botrytis cinerea, Alternaria alternaria 저해균주의 탐색 37
2. 항진균활성을 보유한 균주의 분리 및 동정 41
II. 항진균활성 물질의 분리 및 동정 52
1. 선정된 미생물 균주로부터 항진균물질의 분리 및 동정 52
2. 항진균물질의 분리 및 구조 81
3. 항진균물질 생산을 위한 최적조건의 설정 88
4. 분자적 수준에서의 항진균기전 탐색 102
5. 항진균물질의 안전성 평가 115
6. 항진균물질 처리에 따른 복숭아의 품질평가 131
III. 시제품 생산 및 이에 의한 항진균 효과 검정 143
1. 복숭아의 수확후선도유지제 개발 143
2. 시제품을 처리한 복숭아에서의 곰팡이 발생률 평가 144
3. 시제품을 처리한 복숭아의 품질 평가 145
제4장 목표달성도 및 관련분야에의 기여도 150
1. 복숭아 병원성 진균 저해활성보유균주의 탐색 150
2. 선정 균주로부터 항진균활성 물질의 분리 및 동정 150
3. 항진균활성 물질 처리후 복숭아의 품질 평가 150
4. 항진균활성 물질의 항진균기전 탐색 151
5. 시제품 생산과 항진균 효과 검정 및 복숭아 처리시의 품질 평가 152
6. 연구성과 153
제5장 연구개발결과의 활용계획 154
제6장 연구개발과정에서 수집한 해외과학기술정보 155
1. Aureobasidum pullulans 155
2. Cyptococcus Iaurentii (Kufferath) Skinner 155
3. Metschnikowia fructicola 156
4. Pichia guilliermondii 156
5. 사과의 수확후 곰팡이 조절을 위한 Hurdle concept 156
제7장 참고문헌 157
표 1. 우리나라 복숭아 재배면적 및 생산량 변화 13
표 2. 복숭아 농가의 연도별 소득 변화 14
Table 1. 3-factor rotatable central composite design for optimum condition of phenylethyl alcohol production 31
Table 2. Composition of experimental diets 34
Table 3. Antifungal effect of isolated strains from human feces. 37
Table 4. Carbon compoumds assimilation of selected three strains. 43
Table 5. Levels of partial 26S rDNA sequences similarity for selected three strains and representatives of some other taxa. 45
Table 6. Action of P. farinosa SKM-1, P. anomala SKM-T, and G. geotrichum SJM-59 against the plant pathogene, M. fructicola, R. nigricans, P. expansum, B. cinerea, A. alternaria. 54
Table 7. Characteristics of identified antifungal compounds from pichia farinosa SKM-1. 60
Table 8. Characteristics of antifungal compounds from pichia anomala SKM-T. 66
Table 9. Characteristics of identified antifungal compounds from Galactomyces geotrichum SJM-59. 74
Table 10. Growth and production of phenylethyl alcohol by Pichia anomala SKM-T in media with various carbon substrates 88
Table 11. Growth and production of tryptophol by Pichia anomala SKM-T in media with various carbon substrates 89
Table 12. Growth and production of phenylethyl acetate by Pichia anomala SKM-T in media with various carbon substrates 89
Table 13. Growth and production of phenylethyl alcohol by Pichia anomala SKM-T in media with various L-phenylalanine concentrations 90
Table 14. Growth and production of tryptophol by Pichia anomala SKM-T in media with various L-phenylalanine concentrations 91
Table 15. Growth and production of phenylethyl acetate by Pichia anomala SKM-T in media with various L-phenylalanine concentrations 91
Table 16. The equation for the optimum condition of antifungal compounds production from Pichia anomala SKM-T 95
Table 17. Rotatable central composite design of optimum culture condition for the antifungal compounds production from Pichia anomala SKM-T 96
Table 18. Some different expressions of up-regulated genes in gene control versus antifungal compounds treated groups. 111
Table 19. Some different expressions of down-regulated genes in gene control versus antifungal compounds treated groups. 114
Table 20. Influence on body weight, food intake, and food effciency of single-dose toxicity test 115
Table 21. Influence on the weight of major organs of single-dose toxicity test 116
Table 22. Influence on urine of single-dose toxicity test 117
Table 23. Influence on plasma of single-dose toxicity test 117
Table 24. Influence on EDTA-blood of single-dose toxicity test 118
Table 25. Influence on body weight, food intake, and food efficiency of repeated-dose toxicity test 119
Table 26. Influence on the weight of major organs of repeated-dose toxicity test 120
Table 27. Influence on urine of repeated-dose toxicity test 121
Table 28. Influence on plasma of repeated-dose toxicity test 121
Table 29. Influence on EDTA-blood of repeated-dose toxicity test 122
Table 30. Antimutagenic effect of antifungal compounds against 4-Nitroqulnoline-1-oxide, Sodium azide, and 2-Aminoanthracene in Salmonella typhimurium TA98 and TA100 123
Table 31. Cytotoxic effect on normal murine liver and intestinal cell lines of antifungal compounds 124
Table 32. Relative inhibitory effect on human cancer cell lines of Antifungal compounds 125
Table 33. Effect of antifungal compounds and saline on the stability of RBC membrane 126
Table 34. Effect of antifungal compounds on the activities of hepatotoxicity related enzymes. 127
Table 35. Flavor analysis of Prunus persica treated with PEA, TRTP, and PEAC 139
Fig 1. Antifungal activities of Pichia farinosa SKM-1 on the plant pathogenic fungi, Monilinia fructicola.... 38
Fig 2. Antifungal activities of Pichia anomala SKM-T on the plant pathogenic fungi, Monilinia fructicola.... 39
Fig 3. Antifungal activities of Galactomyces geotrichum SJM-59 on the plant pathogenic fungi, Monilinia fructicola.... 40
Fig 4. Morphology of selected three strains using scanning electron microscopy.... 42
Fig 5. Ascospores of three isolated strains cultured on Kleyn media for 3 day at 25℃.... 44
Fig 6. Partial 26S rDNA sequences of selected three strains.... 48
Fig 7. Phylogenetic tree based on partial 26S rDNA sequences showing the position of strain No. S1 and some other taxa.... 49
Fig 8. Phylogenetic tree based on partial 26S rDNA sequences showing the position of strain No. ST and some other taxa.... 50
Fig 9. Phylogenetic tree based on partial 26S rDNA sequences showing the position of strain No. 59 and some other taxa.... 51
Fig 10. The growth curves of P. farinosa SKM-1, P. anomala SKM-T and G. geotrichum SJM-59.... 53
Fig 11. Chromatograms of antifungal compounds from Pichia farinosa SKM-1, Pichia anomala SKM-T and Galactomyces geotrichum SJM-59.... 56
Fig 12. GC-MS spectrum of identified antifungal compounds from Pichia farinosa SKM-1.... 59
Fig 13. GC-MS spectrum of identified antifungal compounds from Pichia anomala SKM-T.... 65
Fig 14. GC-MS spectrum of identified antifungal compounds from Galactomyces geotrichum SJM-59.... 73
Fig 15. Antifungal activities of separated components from P. farinosa SKM-1, P. anomala SKM-T, ang G. geotrichum SJM-59 on Altanira alternaria of eight days of fermentation. 75
Fig 16. Antifungal activities of separated components from P. farinosa SKM-1, P. anomala SKM-T, ang G. geotrichum SJM-59 on Monilinia fructicola on five days of fermentation. 76
Fig 17. Antifungal activities of separated components from P. farinosa SKM-1, P. anomala SKM-T, ang G. geotrichum SJM-59 on Botrytis cinerea on five days of fermentation. 77
Fig 18. Antifungal activities of separated components from P. farinosa SKM-1, P. anomala SKM-T, ang G. geotrichum SJM-59 on Penicillium expansum on eight days of fermentation. 78
Fig 19. Antifungal activities of separated components from P. farinosa SKM-1, P. anomala SKM-T, ang G. geotrichum SJM-59 on Rhizopus nigricans on eight days of fermentation. 79
Fig 20. Antifungal activities of separated components from P. farinosa SKM-1, P. anomala SKM-T, ang G. geotrichum SJM-59 on Rhizopus nigricans on five days of fermentation. 80
Fig 21. EI-MS spectrum of the first isolated antifungal compound (PEA) from antifungal yeast cultures.... 81
Fig 22. Scanning of the frist antifungal isolate (PEA) from the yeast cultures.... 82
Fig 23. FT-IR spectrum of the first isolated antifungal compound (PEAC) from yeast cultures. 83
Fig 24. ¹H-NMR spectrum (400MHz) of phenylethyl alcohol (PEA) in CDCI₃.... 84
Fig 25. Scanning of the second antifungal isolates (TRTP) from the yeast cultures. 85
Fig 26. FT-IR spectrum of the second isolated antifungal compound (TRTP) from yeast cultures. 85
Fig 27. EI_MS spectrum of the second isolated antifungal compound (TRTP) from antifungal yeast cultures.... 86
Fig 28. ¹H-NMR spectrum (400MHz) of tryptophol (TRTP) in CDCI₃.... 86
Fig 29. FT-IR spectrum of the third isolated antifungal compound (PEAC) from yeast cultures. 87
Fig 30. EI-MS spectrum of the third isolated antifungal compound (PEAC) from antifungal yeast cultures.... 87
Fig 31. Effect of temperature on the growth of Pichia anomala SKM-T.... 92
Fig 32. Effect of various pH on the growth of Pichia anomala SKM-T.... 93
Fig 33. Effect of agitation speed on the growth of Pichia anomala SKM-T.... 94
Fig 34. Normal plot residuals 97
Fig 35. Validity of rotatable central composite design experiments 98
Fig 36. Validity of rotatable central composite design 99
Fig 37. Pertubation of rotatable central composite design experiments 100
Fig 38. Response surface contour map of antifungal compounds production in Pichia anomala SKM-T.... 101
Fig 39. Light microscope of Monilia fructicolar treated with antifungal compounds(cpompounds).... 102
Fig 40. The spore formation of Aspergillus niger treated with antifungal compounds (100 ppm). 103
Fig 41. The mycelium formation of Rhizopus nigricans treated with antifungal compounds (100 ppm). 104
Fig 42. The mycelium formation of Alternaria alternaria treated with antifungal compounds (100 ppm). 105
Fig 43. The melanine formation of Botrytis cinerea treated with antifungal compounds (100 ppm). 106
Fig 44. The spore formation of Alternaria alternaria treated with antifungal compounds (100 ppm). 107
Fig 45. Tne DHN melanogenesis pathway in black fungi. 108
Fig 46. Northern blot analysis of mRNA from alternaria alternaria treated with 100 ppm of TRTP, PEAC, ad PEA.... 109
Fig 47. Liquid fermentation of Alternaria alternaria treated with 100 ppm of TRTP, PEAC, and PEA for 5 days at room temperature. 110
Fig 48. Two dimensional anaysis of Alternaria alternaria treated with 100 ppm of PEA, TRTP, and PEA.... 112
Fig 49. Highly expressed proteins Alternaria alternaria (control) 113
Fig 50. Less epressed proteins in Alternaria alternaria (control) 113
Fig 51. The degree of RBC hemolysis.... 126
Fig 52. Histological examination of various organs extracted from the repeated-dose toxicity testing rat 130
Fig 53. Effects of PEA, TRTP, and PEAC on the fungal decay rate of 'Yumyung', peach fruits for 6 days at room temperature. 131
Fig 54. Effects of PEA, TRTP, and PEAC on the relative weight loss of 'Yumyung', peacth fruits for 6 days at room temperaure. 132
Fig 55. Effects of PEA, TRTP, and PEAC on the index of woolliness of 'Yumyung', peach fruits for 6 days at room temperature. 133
Fig 56. Effects of PEA, TRTP, and PEAC on the relative change of hardness (xg) of `Yumyung`, peach fruits for 6 days at room temperature. 134
Fig 57. Effects of PEA, TRTP, and PEAC on the concentration (mg/g) of ascorbic acid of 'Yumyung', peach fruits for 6 days at room temperature. 135
Fig 58. Effects of PEA, TRTP, and PEAC on the concentration (mg/g) of reducing sugar of 'Yumyung', peach fruits for 6 days at room temperature. 136
Fig 59. Effects of PEA, TRTP, and PEAC on the titratable acidity (malic acid, %) of 'Yumyung', peach fruits for 6 days at room temperature. 137
Fig 60. Effects of PEA, TRTP, and PEAC on ethylene production of 'Yumyung', peach fruits for 6 days at room temperature. 138
Fig 61. Peach-taut, a new postharvest biological control agent using PEA, TRTP, PEAC from extracted from natrual 143
Fig 62. Effect of Peach-taut on the fungal decay rate of 'Hongbak', peach fruits for 10 days at room temperature. 144
Fig 63. Effects of Peach-taut on the relative weight loss of 'Hongbak', peach fruits for 8 days at roon temperature. 145
Fig 64. Effects of Peach-taut on the index of wooliness of 'Hongbak', peach fruits for 8 days at room temperature. 146
Fig 65. Effects of Peach-taut on the hardness of 'Hongbak', peach fruits for 8 days at room temperature. 147
Fig 66. Effects of Peach-taut on the concentration of ascorbic acid of 'Hongbak', peach fruits for 8 days at room temperature. 148
Fig 67. Effects of Peach-taut on the concentration of reducing sugar of 'Hongbak', peach fruits for 8 days at room temperature. 149