와송(Orostachys japonicus A.)의 기내배양과 유용물질의 생리활성에 관한 연구

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Orostachys japonicus, tissue culture, the fraction extract, antioxidant, antibiotic
In the tissue culture using leaf tissue of Orostachys japonicus, callus were induced most efficiently in 5-week-after culture in the MS medium supplemented with 10 μM BAP and 4 μM 2,4-D at the growth condition of 24℃ 6500Lux 16 hr-day-length. Adventitious roots developed in all the medium supplemented with 0, 0.1, 1 μM 2,4-D whereas roots formed only in the uppersides of the leaflets not in contact with MS medium containing 0.5 μM BAP and 100 μM 2,4-D. Shoot regeneration was observed when callus induced in the MS medium containing 10 μM BAP and 4 μM 2,4-D was transplanted in to the MS medium without any plant growth regulators. Massive proliferation methods given by these tissue culture methods for O. japonicus might provide efficient ways to its production.
The ethylacetate fraction extract for O. japonicus contained 634.48 μg/㎖ polyphenol and 205.20 μg/ml flavonoid. The polyphenol and flavonid contents of O. japonicus were 0.16x and 3.3x higher than those of Aronia a source of well-known health supplement food, respectively. The ABTS radical scavenging ability of ethylacetate fraction extract at 1 mg/ml was higher than 95% which is comparable to ascorbic acid of 97%. The APX enzymatic activity and catalase activity were 1125.89 μmol ascorbate oxidized/min/mg protein and 119.87 H2O2 decomposed/ min/mg protein, respectively. In disc agar plate diffusion assay, the extract gave rise to a larger inhibition circle with Lissteria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus and Malassezia furfur strains compared with antibiotics kanamycin suggestive of high antibiotic activity. These results suggest that fraction extract of O. japonicus might be very effective and economical in developing natural antioxidant and antibiotic.
Alternative Title
A study on in vitro culture of Orostachys japonicus A. and physiological activity of extract
Alternative Author(s)
Kim Seung Mi
일반대학원 생명과학
Awarded Date
Table Of Contents
제 1 장 서론 1
제 2 장 재료 및 방법 3
2.1 재료식물 및 시료의 추출 3
2.2 실험 방법 5
2.2.1 Tissue culture 5
가. 생장조절제의 영향 5
나. Callus culture 7
2.2.2 생리활성의 측정 8
가. 폴리페놀과 플라보노이드 함량 측정 8
(1) 총 폴리페놀 함량 측정 8
(2) 총 플라보노이드 함량 측정 9
나. 항산화 활성 능력 측정 10
(1) DPPH radical 소거활성 10
(2) ABTS radical 소거활성 11
(3) 아질산염 소거활성 12
다. 항산화 효소활성 측정 13
(1) 효소액의 조제 13
(2) Catalase (CAT)의 활성 측정 13
(4) Ascorbate Peroxidase (APX)의 활성 측정 13
(5) Peroxidase (POX)의 활성 측정 14
라. Tyrosinase 활성 저해효과 측정 15
2.2.3 항균활성 측정 16
가. 사용 균주 및 배지 16
나. 균주의 배양 및 항균활성 효과 측정 16
2.2.4 항암활성 효과 22
가. 세포의 배양 22
나. 인체 암세포의 생존율 측정 22
2.2.5 통계처리 23
제 3 장 결과 및 고찰 24
3.1 Tissue culture 24
3.1.1 생장조절제의 영향 24
3.1.2 Callus culture and plant regeneration 33
3.2 생리활성 측정 39
3.2.1 총 폴리페놀 함량 및 총 플라보노이드 함량 39
3.2.2 항산화 활성 능력 42
가. DPPH radical 소거활성 42
나. ABTS radical 소거활성 44
다. 아질산염 소거활성 46
3.2.3 항산화 효소활성 48
3.2.4 Tyrosinase 활성저해 효과 53
3.3 항균활성 효과 55
3.4 MTT assay에 의한 인체 암세포 생존율 59
3.4.1 폐암 세포주(Calu-6)의 생존율 60
3.4.2 유방암 세포주(MCF-7)의 생존율 62
3.4.3 대장암 세포주(HCT-116)의 생존율 64
제 4 장 요약 및 결론 67
제 5 장 참고문헌 68
김승미. (2016). 와송(Orostachys japonicus A.)의 기내배양과 유용물질의 생리활성에 관한 연구.
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