곰취(Ligularia fisheri )를 이용한 callus배양 및 항산화 활성 검정

강현욱,김영윤,김추자,박요셉,서완우,양정호,연지영,이소연,이인순,문혜연 대구대학교 산업기술연구소 2011 産業技術硏究 Vol.22 No.1

We described the effects of antioxidant activity on the extracts of tissue and callus formation by Ligularia fisheri . The callus culture was induced from leaf tissue of Ligularia fisheri in both MS and B5 medium with IAA(0∼1.5 ㎎/ℓ) or/and kinetin (0∼1.0 ㎎/ℓ). As medium for callus culture induced from Ligularia fisheri , B5 and MS medium showed no callus formation. Both media brought to decrease differentiation of callus, on the other hand those induced to accelerate cell development. The hytohormones were required over 0.4 ㎎/ℓ of IAA and kinetin for formation of callus and those conditions were induced the pigmental deposit like dark red color in callus tissue. In order to extract functional material from natural leaf and callus tissue of Ligularia fisheri used distilled water and 75% ethanol as solvent and those extract used to determine antioxidant activity as 1, 1-diphenyl-2-picryl-hydrozyl (DPPH) radicals scavenging effect and TBA(thiobarbituric acid) activity as well as the concentration of total phenol and flavonoid. In result, the effect of DPPH and TBA had high activity more callus extract of red color than callus(green color) and natural leaf tissue and it also had high concentration of flavonoid. That is because the high concentration of phytohormone brought stress tolerance and hypersensitivity in callus tissue of Ligularia fisheri , so this reaction induced to change color of callus tissue like pigment deposit for culture.

헛개나무의 캘러스 형성 및 multiple shoot 유기

엄승희(Seung Hee Eom),강원희(Won Hee Kang),신동용(Dong Yong Shin),허권(Kwon Heo),최원철(Won Cheol Choi),이현용(Hyeon Yong Lee),유창연(Chang Yeon Yu) 한국자원식물학회 2002 한국자원식물학회지 Vol.15 No.3

본 연구에서는 헛개나무 조직 배양에서 체세포배 유토 및 식물체 재분화를 위한 적절한 배지, 생장조절물질, 탄소원 그리고 배양 환경을 조사하였으며 대량 번식 시스템을 위해 체세포 배로부터 식물체로 재분화 하기전 세포 동조화와 건실한 유묘 생산에 최적인 배양 조건을 조사하였다. 강원도 양양 헛개나무 자생지에서 채취한 종자를 배배양하여 기내 발아시킴으로 기내 식물체를 얻었으며 이를 본실험에 이용하였다. 기내 발아후 7-14일 된 식물체의 자엽과 잎 절편을NAA를 처리한 배지에 배양했을 때 callus 상태를 거치지 않고 직접 shoot가 유기되었는데 NAA 0.5mg/에서 43.6%로 형성율이 가장 높았으며 절편체 당 shoot의 수도 2.8개였다. NAA와 BA 조합처리 하였을 때 BA 0.1mg/ + NAA 1mg/에서 38.1%의 형성율을 보였으며 절편체 당 shoot의 수는 4개 이상으로 유기 되었다. 체세포배는 NAA와 2,4-D의 단독처리 또는 BA와의 조합처리에서 발생하였으며 직접 체세포배 또는 배발생 캘러스가 형성하였다. BA 0.1mg/ + 2,4-D 1mg/ 또는 BA 0.1mg/ + NAA 1mg/ 에서 발생한 배발생 캘러스의 생장과 성숙 및 발아에 최적 배양 조건을 조사하였을 때 GA3 1mg/ 을 처리하여 배 발아를 촉진시켰으며 자엽은 발달하지 못하고 하배축이 신장하여 유근이 발생한 형태의 유묘를 생산하였다. Loaves, stems, cotyledons, and roots of Hovenia dulcis Thunb grown in test tube were cultured on media containing different concentrations of single or combined growth regulators. In MS media containing 2mg/ι BA, the shoot formation rate was 95.5% and it was the highest frequency of shoot formation. MS media showed most efficiency in the shoot formation at 0.01mg/ι TDZ for the callus formation, but the color of callus changed to brown at a higher concentration of TDZ. Callus formation was 89.% at 0.5mg/ 2.4-D, but IAA, IBA, and NAA were not effective on the formation of callus. Calli were formed only on wound area when IAA, IBA, and NAA were added into MS media. Combined growth regulators (BA + auxin) were more effective in roots and nodes than leaves and cotyledons on the formation of shoot. More than 97% of shoot formation was obtained on MS media containing BA and auxin. For the production of multiple shoot, nodes of Hovenia dulcis were used and effect of growth regulators on the formation of multiple shoot was evaluated on MS media. Highest shoots (5.3) of Hovenia dulcis were induced on MS media supplied with 0.1mg/ι BA and 0.1mg/ι NAA, and an average of 6.4 shoots per explant were obtained in 1/2 MS media containing same concentration and growth regulators. An average of 7 shoots per explant after 4 weeks of culture from nodes of Hovenia dulcis was produced on a woody plant medium(WPM) containing 0.1mg/ι BA and 0.1mg/ι NAA. Shoot length was 6.0 cm in average.

고려 엉겅퀴(Cirsium setidens Nakai) 잎 조직을 이용한 callus 배양 및 항산화 활성 검증

박정훈,심예지,박기임,이인순,문혜연 대구대학교 특수교육재활과학연구소 2010 産業技術硏究 Vol.21 No.1

We described the effects of antioxidant activity on the extracts of tissue and callus formation by Cirsium setidens Nakai. The callus culture were induced from leaf tissue of Cirsium setidens in both MS and B5 medium with IAA or/and kinetin (0∼5.0 ㎎/ℓ). As medium for callus culture induced from Cirsium setidens, B5 medium showed better condition than MS media. MS medium brought to decrease callus differentiation after formation, on the other hand B5 medium induced to accelerate callus differentiation. Among phytohormones is required 0.5 ㎎/ℓ of IAA and between 2.0 ㎎/ℓ and 1.0 ㎎/ℓ of Kinetin for formation of callus. In order to extract functional material from natural leaf and callus tissue of Cirsium setidens used distilled water and 75% ethanol as solvent and those extract used to determine antioxidant activity as alpha-diphenyl-2-picryl-hydrozyl (DPPH) radicals scavenging effect and TBA(thiobarbituric acid) activity. In result, the effect of DPPH and TBA had high activity more natural leaf extract than callus tissue and 75% ethanol was solvent better than distilled water. The 75% ethanol extract of natural leaf showed an excellent antioxidant activity because that particularly had highest concentration of vitamin C and total phenol among those extracts

잔디의 캘러스 형성 및 재분화에 끼치는 식물생장조절제, 탄소원 및 AgNo₃의 영향

임요섭,한성수,정재훈 圓光大學校大學院 1996 論文集 Vol.15 No.-

잔디의 형질전환체계를 확립하기에 앞서 효율적인 재분화 체계를 확립하고자 식물생장 조절제, 탄소원 및 AgNO₃가 재분화에 미치는 영향을 조사한 결과는 다음과 같다. 1. 배지(MS와 N6) 및 식물생장조절제(2,4-D와 NAA)에 따른 Korean lawngrass(Zoysia japonica Steud.)와 penncross creeping bentgrass (Agrostis palustris Huds.)의 캘러스 유기는 두 품종 모두 N6배지에서 보다 MS배지에서 좋았으며, Korean lawngrass는 2,4-D 1mg/L 첨가 MS배지에서 그리고 penncross creeping bentgrass는 2,4-D 2mg/L첨가 MS배지에서 가장 좋았다. 2. 캘러스증식은 두 품종 모두 2,4-D 2mg/L함유 MS배지에 NAA 0.2mg/L첨가하였을 때 가장 좋았고, kinetin을 Korean lawngrass의 경우 0.2mg/L, penncross creeping bentgrass의 경우 0.3mg/L를 첨가하였을 때 가장 좋았다. 3. 식물체 재분화 실험에서 Korean lawngrass 잎 절편체의 재분화율은 MS배지에서 2,4-D 2mg/L+NAA 0.2mg/L+kinetin 0.1mg/L 조합처리시 44% 그리고 stock callus의 재분화율은 2,4-D 2mg/L+NAA 0.2mg/L+kinetin 0.3mg/L 조합처리시 32%이었다. penncross creeping bentgrass의 잎 절편체와 stock callus의 재분화율은 kinetin 0.3mg/L 농도에서 각각 80%와 67%이었다. 4. Korean lawngrass의 경우 전술한 재분화 배지에 AgNO₃ 1mg/L를 첨가하였을때 3∼4% 그리고 pencross creeping bentgrass의 경우 2mg/L를 첨가하였을때 10∼16%의 재분화율이 향상 되었다. This study was carried out to determine the effects of growth regulators, carbon sources and silver nitrate on callus formation and plant regeneration of Turfgrass. The results were summarized as fallows : 1. Callus from Korean lawngrass (Zoysia japonica Steud.) and pencross creeping bentgrass (Agrostis palustris Huds.) induced better in MS medium than in N6 medium and by addition of 2,4-D than by that of NAA. Callus formation from Korean lawngrass and penncross creeping bentgrass was very effective at MS medium adding 1mg/L 2,4-D and at the medium adding 2mg/L 2,4-D, repectively. 2. Growth of callus was good at MS medium containing 2mg/L 2,4-D + 0.2mg/L NAA. Callus growth of Korean lawngrass and penncross creeping bentgrass was good when kinetin was added 0.2mg/L and 0.3mg/L, individually, to MS medium containg 2mg/L 2,4-D + 0.2mg/L NAA. 3. Regeneration rate from leaf and stock callus of Korean lawngrass was 44% at MS medium adding 2,4-D 2mg/L + NAA 0.2mg/L + kinetin 0.3mg/L and 32% at the medium containing 2,4-D 2mg/L + NAA 0.2mg/L + kinetin 0.3mg/L, each and that from leaf and stock callus of penncross creeping bentgrass was 80% and 67%, each, at the medium adding 2,4-D 2mg/l + NAA 0.2mg/L + kinetin 0.3mg/L. 4. Regeneration rate of Korean lawngrass and penncross creeping bentgrass increased by 3 to 4% and by 10 to 16%, respectively, when added AgNO₃ 1∼2mg/L to the above-mentioned regeneration medium.

브로콜리 줄기의 callus배양 및 항노화 효과

박태진 ․ 이상민 ․ 조준환 ․ 남수호․ 신주은 ․ 김대환 ․ 이인순 ․ 박흠대 대구대학교 산업기술연구소 2018 産業技術硏究 Vol.29 No.2

Broccoli (Brassica oleracea var. italica) is well known of excellent antioxidant food so that we researched on its anti-aging and callus differentiation from the tissue of broccoli stem by MS medium with exo-phytohormone as IAA, kinetin. The functional materials of callus tissue were extracted with water and 70% EtOH after 16 days progressed callus formation. As a result, callus differentiation and formation were showed higher efficiency at B5 condition medium (0.1 mg/L IAA, 0.4 mg/L). The callus cultured on B3, B6 and B7 medium were observed the reaction of enzymatic browning to callus tissue. The anti-aging effect on the extract was experiment with DPPH radical scavenging and catalase(CAT). The activity of DPPH radical scavenging was showed high value in water extract of B1, B6, B8, B9 and was increased depending on sample concentration. However, in case of the extract of 70% EtOH, DPPH activity was showed different effect depending on the sample kind. CAT activity was found the highest enzyme unit in the extract of natural broccoli and B1(no add exo-phytohormone). It is to express that a broccoli itself is great food for anti-aging. Therefore, callus formation of broccoli considers having no economic efficiency to do development of anti-aging product.

Nitrogen Stress가 적양배추(Brassica oleracea)의 callus분화 및 Anthocyanin 함량변화와 항산화 활성에 미치는 효과

예다인,김혜림,김효정,최문정,김초록,김영지,이지현,구민성,정 강현,이인순,문혜연 대구대학교 산업기술연구소 2012 産業技術硏究 Vol.23 No.1

In order to study on the effects of antioxidant activity, changing concentration of anthocyanin and callus formation of Brassica oleracea by nitrogen stress experimented callus culture in MS medium with 0.1 ㎎/ℓ IAA, 0.2 ㎎/ℓ kinetin and 0∼5 mM KNO3. As medium showed differentiation of callus and the pigmental deposit like red color inside callus tissue. We extracted functional material from natural leaf and callus tissue of Brassica oleracea to use 70% ethanol as solvent and those extract used to determine antioxidant activity as 1, 1-diphenyl-2-picryl-hydrozyl (DPPH) radicals scavenging effect and nitric oxide radical scavenging effect as well as the concentration of total phenol and flavonoid. Additionally, we measured the concentration of anthocyanin and catalase activity from natural and callus tissue. In result, M1 had over 35% anthocyanin concentration, 1.7 times catalase activity and 20% DPPH activity compared with control. MC, M2 and M3 also showed 50% DPPH activity compared with positive control, BHT. However, more natural tissue than callus tissue cultured with nitrogen source had high effect on nitric oxide radical scavenging and the concentration of phenol compound, flavonoid and vitamin C. As a result, nitrogen source brought hypersensitivity in callus tissue of Brassica oleracea, so this reaction only induced to change color of callus tissue, pigment deposit for culture and also increase anthocyanin concentration, DPPH activity. Therefore, natural tissue of Brassica oleracea is good food for anticancer and detoxification and it has high concentration of functional compound.

무궁화 품종 '난파'의 효율적인 재분화 체계 확립

손지희,선현진,강홍규,서석철,이효연,Son, Ji-Hi,Sun, Hyeon-Jin,Kang, Hong-Gyu,Suh, Seok-Chul,Lee, Hyo-Yeon 한국식물생명공학회 2019 식물생명공학회지 Vol.46 No.4

Hibiscus syriacus L., the national flower of Korea, is a commonly grown ornamental species found in parks, gardens and along roadsides in Korea. This study sought to establish a plant regeneration system of H. syriacus 'Nanpa' using mature leaves as an explant. Sterilized mature leaf explants were cultured on modified MS medium with combinations of NAA and BAP at various concentrations for 6 weeks. Among the various media evaluated, modified MS media containing 0.46 mg/L NAA and 0.5 mg/L BAP (CI-1) or 0.92 mg/L NAA and 1 mg/L BAP (CI-4) were the most effective for callus formation. Whitish-yellow calluses were observed on CI-1 medium, while green calluses formed on CI-4 medium. The whitish-yellow callus derived from CI-1 medium showed a higher frequency of shoot induction than the green callus derived from CI-4 on modified MS medium containing 0.5 mg/L BAP. Among the various media evaluated in this study, modified MS medium containing 0.46 mg/L NAA and 0.5 mg/L BAP was the most optimal for shoot-forming callus production. Our findings show that mature leaves of H. syriacus 'Nanpa' can be used for in vitro plant regeneration, and the regeneration system described here may be a powerful tool for molecular breeding of H. syriacus 'Nanpa' in the future.

왜성 변이 품종 ‘꼬마’ 무궁화의 캘러스 유도 및 지상부 형성에식물생장조절물질이 미치는 영향

이지연,강은정,김상훈,김동섭,김진백,하보근,강시용 한국방사선산업학회 2011 방사선산업학회지 Vol.5 No.3

‘Ggoma’ is a new Hibiscus dwarf type variety developed by gamma irradiation at theKorea Atomic Energy Research Institute (KAERI). This study was conducted to determine thebest optimum cultural callus formation and shoot regeneration condition. Sterilized leaf tissueswere cultured on MS (Marashige and skoog’s) medium containing 3% sucrose, 0.8% agar withdifferent concentration and combination of TDZ, 2, 4-D, KT, BA, and 2iP for 4 weeks in vitro culture. MS medium containing 2,4-D 0.1 mg l-1 and BA 0.5 mg l-1 were most effective on callusformation and growth. After 4 weeks, callus was transferred on BA (0.5, 1, 1.5 mg l-1) and TDZ(0.1, 0.2, 0.3 mg l-1) for shoot formation. The best condition for inducing the shoot from callus wasBA 1.5 mg l-1 and TDZ 0.3 mg l-1. This result will be useful for the rapid multiplication of Hibiscussyriacus L.var. Ggoma.

뽕나무 癒合促進 古條揷木에 關한 硏究 : Ⅱ.揷木溫度가 發根生長에 미치는 影響 Ⅱ.Effect of Callusing Temperature on Root Formation and Growth

金浩樂,崔勝雲,林秀浩 한국잠사학회 1991 한국잠사곤충학회지 Vol.33 No.2

Mulberry cuttings from shoots of Shinkwangppong (Morus bombycis koidz.) had been callused in vermiculite separately at 15, 20, 25 and 30°C for 15 days before transplanting them in greenhouse to make clear the effect of temperature on root formation and growth is as follows. The buds of cuttings started sprouting in 4 and 6 days of callusing at 30 and 25°C, respectively, reaching 100% budding in 10 and 15 days of callusing. Budding was delayed, however, at low temperature, showing 86% and 92% at 15 and 20°C, respectively, in 15 days. Rooting from the cuttings was also accelerated at high temperature, showing 97~ 100% rooting at 25°C and 30°C, in 15 days of callusing but no more than 93% at low temperature even in 35 days. Although high temperature increased root number and length after 15 days in callusing, no differences showed in the number and the weight at more than 20°C in 35 days of cutting.

무궁화 품종 ‘난파’의 효율적인 재분화 체계 확립

손지희,선현진,강홍규,서석철,이효연 한국식물생명공학회 2019 JOURNAL OF PLANT BIOTECHNOLOGY Vol.46 No.4

Hibiscus syriacus L., the national flower of Korea, is a commonly grown ornamental species found in parks, gardens and along roadsides in Korea. This study sought to establish a plant regeneration system of H. syriacus ‘Nanpa’ using mature leaves as an explant. Sterilized mature leaf explants were cultured on modified MS medium with combinations of NAA and BAP at various concentrations for 6 weeks. Among the various media evaluated, modified MS media containing 0.46 mg/L NAA and 0.5 mg/L BAP (CI-1) or 0.92 mg/L NAA and 1 mg/L BAP (CI-4) were the most effective for callus formation. Whitish-yellow calluses were observed on CI-1 medium, while green calluses formed on CI-4 medium. The whitish-yellow callus derived from CI-1 medium showed a higher frequency of shoot induction than the green callus derived from CI-4 on modified MS medium containing 0.5 mg/L BAP. Among the various media evaluated in this study, modified MS medium containing 0.46 mg/L NAA and 0.5 mg/L BAP was the most optimal for shoot- forming callus production. Our findings show that mature leaves of H. syriacus ‘Nanpa’ can be used for in vitro plant regeneration, and the regeneration system described here may be a powerful tool for molecular breeding of H. syriacus ‘Nanpa’ in the future.