홍경천(Rhodiola rosea L.)의 액아배양을 통한 다신초 유도 및 기내 대량증식

배기화,배기화,김남영,송재모,고명석,송관필 한국식물생명공학회 2012 식물생명공학회지 Vol.39 No.2

An efficient in vitro propagation was established by using axillary bud explants of roseroot (Rhodiola rosea L.), which has been known as a medicinal plant in East Asia. Among various media tested, MS medium supplemented with 1.0 mg/L BA and 1.0 mg/L GA3 was found to be the best for multiple shoot formation (15 axillary shoots per axillary bud). In addition 1/2MS medium containing 50 g/L sucrose was best for shoot elongation (7.8 cm) and increasing total chlorophyll contents (8.64 mg/g) best. Maximum number of roots (17.7 roots per explant) was observed on the medium without plant growth regulators. Propagated plants were successfully acclimatized to ex vitro conditions, with a survival frequency of 97% after 12 weeks. Most rooted shoots grew well and produced viable seeds when grown in vitro culture conditions. Therefore, R. rosea can be effectively propagated in vitro by the system we developed in this study. 본 실험은 약리적 효능이 뛰어난 홍경천의 기관 (액아가포함된 줄기 조직)배양을 통한 대량증식 방법의 일환으로 액아배양을 통한 다신초를 유도, 증식조건에 관해 BA 와 GA3의 영향을 조사하였고 신초증식에 필요한 적정sucrose의 농도를 확인하였다. 그 후 적절한 기내 발근 및토양순화 조건을 탐색하여 최적의 홍경천의 생산 조건을확립하였다. 다신초의 유도 및 증식은 GA3와 BA를 혼합처리가 효과적이었다. 신초증식에 효과적인 sucrose의 농도는 50 g/L가 첨가된 배지였으며, 평균 7 cm 이상의 신장을 보였다. 기내 발근은 옥신류 식물생장조절물질 무첨가 배지에서 가장 높은 발근길이와 개수를 나타냈다. 기내발근된 유식물체의 토양순화는 모래상을 제외하고는 모두 양호한 생장을 보였고, 특히 모래, 피트모스, 펄라이트가 모두 첨가된 배합토에서 가장 양호한 생장을보였다. 이상의 결과는 수년전 국내에 도입하려다 실패한 약용식물인 홍경천의 기내배양을 통한 재배의 가능성을 제시하는 것이고 더 나아가 다양한 생물소재의 활용방안의 기초재료로 활용될 것으로 보여진다.

기내배양을 통한 홍경천(Rhodiola sachalinensis)의 부정근 생산

배기화,윤의수,최용의,Bae, Kee-Hwa,Yoon, Eui-Soo,Choi, Yong-Eui 한국자원식물학회 2009 한국자원식물학회지 Vol.22 No.4

홍경천의 잎, 줄기, 뿌리 절편을 이용하여 기내 부정근의 생산 체계를 확립하였다. 먼저 홍경천의 잎, 줄기, 뿌리 절편을 0.1, 1.0, 3.0 및 5,0 mg/L의 IBA와 sucrose가 10, 30, 50 및 100 g/L가 첨가된 MS 배지위에 치상하여 부정근의 유도율을 조사하였다. 부정근의 유도는 잎, 줄기 절편에서 IBA의 농도가 5.0 mg/L 일때 가장 높은 유도율을 보였으며, 뿌리 절편은 IBA 3.0 mg/L 첨가된 배지에서 부정근 유도율이 가장 높았다. Sucrose의 농도는 30 g/L가 첨가되었을 때 잎, 줄기, 뿌리 절편에서 높은 유도율을 나타났다. 고체배지 조건에서 부정근의 유도율이 가장 우수한 조건을 기본으로 액체배양을 실시하였으며, 염의 농도에 따른 부정근의 증식조건을 조사하였다. 1/3MS 배지에서 홍경천의 부정근을 배양하였을 때 1/2MS, MS 액체 배지조건 보다 약 2배, 2.5배의 부정근 생장량을 보였다. Rhodiola sachalinensis is one of the most popular oriental medicines in East Asia. It is a perennial herb, belonging to the family Crassulaceae, which is mainly distributed in mountains at the altitudes of 1700-2500 m in Baek-Du mountain. Cultivation of this species for the production of medicine materials is not easy in nature, because of restrict habitats. In vitro production of roots can be applicable for the production of medicinal materials. Here, we investigated the optimal conditions for induction and proliferation of adventitious roots in in vitro culture system. Leaf, stem and root segments from R. sachalinensis were cultured on Murashige and Skoog(MS) medium supplemented with the various concentrations of IBA(Indole-3-butyric acid)(0.5, 1.0, 3.0, and 5.0 mg/L) and sucrose(10, 30, 50, and 100 g/L). The optimal explant for adventitious root induction was leaf segment. Induction of adventitious roots was highest on MS medium supplemented with 5.0 mg/L IBA and sucrose 30 g/L. In liquid culture, fresh weight of adventitious roots was highest(15.65 g) on 1/3 strength MS liquid medium supplemented with 5.0 mg/L IBA with 30 g/L sucrose. These results revealed the first attempt for the production of adventitious roots in R. sachalinensis.

조경수종인 소엽맥문동(Ophiopogon japonicus (L.f.) Ker Gawl)의 캘러스 유도 및 식물체 재분화

배기화,윤의수 한국자연보호학회 2013 한국자연보호학회지 Vol.7 No.1

A method for rapid micropropagation of Ophiopogon japonicus through plant regeneration from leaf, rhizome and root explants derived calli has been developed. Leaf, rhizome and root segments were cultured on Murashige and Skoog (MS) medium supplemented with plant growth regulators (2,4-D, NAA, IAA; 0~3.0 mg/ L) for callus induction. Callus production was highest at 1.0 mg/L 2,4-D where 91%. These calli were transferred to MS medium supplemented with various concentrations of 2,4-D (0, 0.5, 1.0, 3.0 mg/L) in combination with 6-benzyladenine (BA: 0, 0.5, 1.0 and 3.0 mg/L) for adventitious shoot regeneration. The addition of low concentration of 2,4-D into BA containing medium significantly increased the frequency of shoot regeneration in both leaf, rhizome and root derived calli. The highest frequency of adventitious shoot (88%) formation was on MS medium supplemented with 0.5 mg/L 2,4-D and 1.0 mg/L BA. For rooting of the shoots, half-strength MS medium supplemented with different concentrations of plant growth regulators (2,4-D, IBA, NAA) 0, 0.1, 1.0, 3.0 mg/L was tried. The optimal results was observed on half-strength MS medium supplemented with 3.0 mg/L NAA (average of 42.9 roots per explant). In vitro raised plantlets were acclimatized and transferred to soil with 100% success. This in vitro propagation protocol would be useful for conservation as well as mass propagation of this medical plant.

대용량 안전밸브의 진동특성에 대한 Moving Mesh를 이용한 해석적 기법 연구

배기화,이우수,김정환,박재현,김정렬,남택근 한국마린엔지니어링학회 2009 한국마린엔지니어링학회 학술대회 논문집 Vol.2009 No.-

Numerical simulations are performed on the compressible air flow through the relief valve (ø 160 ㎜) with the initial room pressure (0.11bar or 0.12bar ) higher than the cracking one (0.1 bar). The numerical method has been validated by comparing the results with the empirical ones. Results show that the disc motion and flow characteristics can be successfully simulated using the moving-mesh strategy and depend strongly on the spring stiffness and the flow passage shape. With increasing spring stiffness, the maximum disc displacement decreases and thus the total disc-opening time also decreases.

Callus induction and plant regeneration of Lychnis wilfordii (Regel) Maxim a critically endangered plant in Korea

배기화,윤의수,Mi Hyun Lee,최용의 한국식물생명공학회 2014 식물생명공학회지 Vol.41 No.1

Lychnis wilfordii (Regel) Maxim is a rare andvalued ornamental plant. Germination rate reached 46.6%when seeds were treated with 100 mg‧l-1 GA (Gibberellicacid)3. The highest callus induction was observed in the leafexplants of the seedlings on MS medium containing specificconcentrations of 0.5 mg‧l-1 BA (N6-benzyladenine) and 3.0mg‧l-1 NAA (a-naphthalene acetic acid). The adventitiousshoot was formed in 97.3% of calli on 1/2 WPM (Woody PlantMedium) medium. Shoot elongation of in vitro propagatedplantlets was no difference among various medium. Theplantlets grew well after transferring to the pot. This in vitropropagation protocol should be useful for conservation ofthis endangered plant.

NaOCl 처리에 의한 한라천마(Gastrodia verrucosa Blume)와 애기천마 (Hetaeria sikokiana Tuyama)의 기내발아

배기화,고명석,최선아,이학봉,김남영,송재모,송관필 한국식물생명공학회 2012 식물생명공학회지 Vol.39 No.3

본 실험은 TTC테스트를 통해 난과식물인 한라천마(G. verrucosa Blume)와 애기천마(H. sikokiana)의 종자기내배양시 NaOCl의 적정 처리시간과 농도조건을 확립한 결과이다. TTC테스트를 통한 종자활력은 애기천마의 경우무처리구에서 95%이상 높았지만 배비대율, 비대배 직경과의 상관관계를 분석해보면 1%에서 30분간 처리 할 경우 종자활력대비 배비대율 77%, 배직경 33 μm로 가장 높게 나타났다. 한라천마 역시 종자활력, 배비대율, 배직경비대, 프로토콤 형성, 프로토콤직경 비대의 상관관계를분석하면 1% NaOCl을 30분간 처리할 경우 최종적으로종자활력대비 배비대율 87%, 배직경 59 μm, 프로토콤의형성율이 91%, 프로토콤 직경 138 μm로 다른 처리구에비해 높게 나타났다. 이상의 결과는 국내외적으로 연구된 바 없는 애기천마와 한라천마의 종자 기내배양 시 발아율을 개선하기 위한 중요한 기초자료로 활용될 것이고더 나아가 자생 생물소재를 활용한 연구의 소재확보의가능성을 제시하는 결과이다. An optimization in vitro seed germination was established by using triphenol tetrazolium chloride (TTC) test, which has been known as two rare orchids (Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama) in Jeju island. We have established proper NaOCl treatment for in vitro germination of G. verrucosa and H. sikokiana through TTC test. In the case of H. sikokiana, seed viability through TTC test was high with 95% in control. However, NaOCl 1% treatment for 30 minutes showed the highest embryo swelling rate to seed viability. Likewise, swelling formation of embryos, diameter of embryos, protocorm formation and diameter of protocorms of G. verrucosa was 87%, 59 μm, 91% and 138 μm through NaOCl 1% treatment for 30 minutes. This result will be applied on the basic information for improving in vitro seed germination rate of G. verrucosa and H. sikokiana.

멸종위기 수종 석곡(Dendrobium moniliforme)의 Protocorm-like Body 형성 및 기내증식

배기화,김남영,송재모,송관필 강원대학교 산림과학연구소 2014 Journal of Forest Science Vol.30 No.1

This experiment was conducted to establish the in vitro propagation of Dendrobium moniliforme through protocorm like body (PLB) induction from the culture of leaf, stem and root explants. Frequency of PLB formation from root explants was higher than those of leaf and stem explants. PLB’s proliferation in MS medium including 1.0 mg/L NAA was better than 1.0 mg/L IBA. Frequency of PLB’s proliferation on medium with various concentrations of BA, Kinetin, Tidiazuron (0. 1.0, 3.0 mg/L) and NAA (0, 1.0 mg/L) was tested. The maximun proliferation of PLB’s was obtained on MS medium supplemented with 3.0 mg/L BA and 1.0 mg/L NAA after 6 weeks of culture. Addition of 200 mg/L activated charcoal (AC) and 30 g/L sucrose was effective on PLB proliferation for D. moniliforme.

멸종위기종인 솔나리(Lilium cernum Komarvo.)의 인편 유래 캘러스 유도 및 기내 식물체 재분화

배기화,윤의수 한국식물생명공학회 2013 식물생명공학회지 Vol.40 No.2

Lilium cernum Komarvo. is an important endangered plant belonging to the family Liliaceae. A method was developed for the rapid micropropagation of L. cernum through plant regeneration from bulb scales explant-derived calli. The bulb scales segments were cultured on Murashige and Skoog (MS) medium supplemented with 0, 0.5, 1.0, 3.0 mg․L-1 kinetin and 0, 0.1, 0.5, 1.0 mg․L-1 NAA or 2,4-D for callus induction. In media with 0.5∼3.0 mg․L-1 kinetin and 0.1∼1.0 mg․L-1 NAA and 2,4-D, 95∼100% of explants produced callus. They were then transferred to MS medium supplemented with various concentrations of NAA (0, 0.01, 0.05 and 1.0 mg․L-1) in combination with BA (0, 1.0 and 2.0 mg․L-1) for bulbet formation. Bulbet induction (78%), weight (468 mg) and size (15.5 mm) were obtained the highest on MS medium containing 2.0 mg․L-1 BA and 1.0 m g․L-1 NAA. In vitro frequency of plant regeneration was not significantly treated in strength of MS and sucrose concentration. Chlorophyll contents in 1/2MS with 50 g․L-1 sucrose treatments were higher than those in control and another treatment. This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.

Plant regeneration from callus of Iris odaesanensis Y. N. Lee native to Korea via organogenesis

배기화,윤의수,유경화,이미현,정재훈,최용의 한국식물생명공학회 2013 식물생명공학회지 Vol.40 No.3

Iris odaesanensis Y. N. Lee. is an important endangered and native plant belonging to the family Iridaceae in Korea. This study describes a method for rapid micropropagation of this species via from leaf, rhizome and root explants derived calli. Leaf, rhizome and root explants were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) for callus induction. Rhizome explants yielded calli at a frequency of 72% when cultured at 1.0 mg/l 2,4-D. Calli were maintained at 1.0 mg/l 2,4-D. These calli were transferred to MS medium supplemented with 0, 0.5, 1.0, and 2.0 mg/l 2,4-D in combination with 0, 0.5, 1.0, and 3.0 mg/l BA for adventitious shoot induction. The highest number of adventitious shoot (228.9 per petri-dish) were formed at 1.0 mg/l 2,4-D and 1.0 mg/l BA. WPM medium was the best to convert calli into plantlets, where up to 98.2% of calli were regenerated into plantlets. This in vitro propagation protocol should be useful for conservation of this endangered plant.

Effects of NaOCl treatment on in vitro germination of seeds of a rare endemic plant, Oreorchis coreana Finet

배기화,송관필,Myoung Suk Ko,Mi Hyun Lee,Nam Young Kim,송재모 한국식물생명공학회 2013 식물생명공학회지 Vol.40 No.1

Oreorchis coreana Finet is threatened globally by over-collection from its natural habitats for horticultural purposes. Its rarity in nature makes this plant one of the most endangered species in Korea. In this study, we investigated the effects of sodium hypochlorite (NaOCl) on orchid seed viability and seed germination. An in vitro bioassay swelling test using immature seeds was compared with a standard chemical procedure using triphenyl tetrazolium chloride (TTC) to test seed viability. In general, the bioassay was more appropriate for estimating embryo viability after a prolonged pre-treatment (more than 1 h) in 1% NaOCl, a surface sterilant often used to enhance germination of seeds of terrestrial plants. Therefore, an efficient method for investigating in vitro swelling of immature seeds is urgently needed. We established a method for determining the viability and swelling of O. coreana seeds via in vitro examination of immature seeds. Treatment of immature seeds with 1% NaOCl for 10 min greatly enhanced the extent of swelling of immature zygote embryos when compared to untreated seeds. These data obtained here appear to be comparable to viability and swelling that occurs in O. coreana seeds via asymbiotic germination.