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Alocasia amazonica의 생물반응기 배양에서 배지 공급 방식이 식물체의 생장과 잎조직 및 기공의 특성에 미치는 영향
백기엽,조은아,한은주 한국식물생명공학회 2006 JOURNAL OF PLANT BIOTECHNOLOGY Vol.33 No.2
Comparative studies on medium supply in bioreactors (raft, immersion and ebb and flood) have revealed that multiplication and growth of Alocasia Amazonica were greatest in the raft system, while lowest in ebb and flood system. In the raft system, the basal part of the shoots was continuously in contact with medium, which enabled a constant uptake of nutrients as well as aeration to the explants. The number and the size of leaf stomata were higher in the raft system compared with immersion and ebb&flood system. In the immersion system, plantlets were deformed and epidermal cells in leaves were irregular with a large intercellular space. The results suggested that the medium supply should be controlled properly to maintain normal and healthy plantlets during liquid cultures in bioreactors which affects morphology and physiology of the plantlets.
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식물생장조절물질, 무기물 농도 및 질소원 비율이 Gymmema sylvestre 세포 배양에 미치는 영향
백기엽,이은정 한국식물생명공학회 2006 JOURNAL OF PLANT BIOTECHNOLOGY Vol.33 No.2
This study was carried out to investigate the effects of plant growth regulators, medium salt strength and nitrogen ratio on cell culture of Gymnema sylvestre. Cell growth was inhibited by 2,4-D concentrations higher than 1.0 mg L-1, but not by kinetin concentrations lower than 0.5 mg L-1. Maximal cell growth was obtained at 1.0 mg L-1 2,4-D and 0.1 mg L-1 kinetin. Cell growth was greatest at 1x MS medium but high strength MS medium inhibited cell growth due to low water potential in the medium. In NH4+:NO3- ratio of 0:60 (ie. 0.0 mM NH4+ and 60 mM NO3-), cells growth was highest but cells were smaller and whiter compared with those in other NH4+:NO3- ratio. Reduced cell growth was observed with continuous culture. These results suggested that optimal culture of Gymnema sylvestre could be achieved with 1x MS medium with 20:40 ratio of NH4+:NO3- supplemented with 1.0 mg L-1 2,4-D and 0.1 mg L-1 kinetin.
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오창호,이태수,이상선,백기엽 충북대학교 한국과학재단 지정 첨단원예기술개발 연구센터 1998 연구보고서 Vol.3 No.-
한국의 춘란 자생지의 춘란(Cymbidium goeringii) 뿌리에서 공생하는 10개의 균근균을 분리하였다. 분리된 난 균근균을 PDA에서 배양한 결과 균총은 다양한 색깔을 나타내었으며, 생장 속도도 달랐다. 분리된 난 균근균을 현미경으로 관찰한 결과 Rhizoctonia repens 또는 R. endophytica var endophytica로 동정되었다. R. repens는 자생란인 춘란의 뿌리에서 많이 분리되었으며, R. endophytica var endophytica는 상업적으로 재배되는 외국산의 난 뿌리에서 분리되었다. 분리된 균들 중에 R. repens와 유사한 5개의 분리균을 oatmeal agar에 접종하고 배양한 후 여러종의 난 유묘를 이식하여 난의 생장을 관찰하였다. 그 결과 난의 생장은 분리 균에 따라서 다양하였으나 대부분의 난은 생장촉진 반응을 나타내서, 분리균과 각각의 난과의 공생 관계를 관찰할 수 있었다. Ten isolates of the orchid mycorrhizal fungi were isolated from the roots of Korean native orchid plants (cymbidium goeringii) which inhabitate mainly in southern and western areas of Korea. The growth rates and color of the isolates in potato dextrose agar (PDA) were various. Microscopic observations of the hyphae isolated were identified as Rhizoctonia repens and R. endophytica var endophytica or their related species. R. repens was isolated from the roots of the Korean native orchids, but R. endophytica var endophytica was only isolated from the roots of the commercial orchids introduced from foreign countries. Also, the polymorphic patterns of genomic DNA extracted from selected isolates were compared with those of DNA extracted from the orchid mycorrhizal fungi isolated previously and similar band patterns were observed among those isolates. Five isolates of R. repens were selected and cultured at the oatmeal agar for investigating their symbiosis with orchid plants. The symbiotic specificity between orchid plants and isolated orchid mycorrhizal fungi was observed by growing orchids about six months in the greenhouse. The symbiotic responses of the commercial orchid plants with selected isolates were quite different from different isolates due to the genetic variations.
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백기엽,유광진,박상일 한국식물생명공학회 1998 식물생명공학회지 Vol. No.
정단 및 마디조직을 이용한 지황의 기내증식방안을 마련하기 위하여 일련의 시험을 실시하였다. 재생된 신초의 정단을 재배양하였을 경우 BA 5.0 mg/L에서 7.8개의 신초가 형성되었으며 투명화된 묘의 발생은 Gelrite 보다는 Bacto-agar 첨가구에서 감소하였다. 광도와 한천의 농도가 증가할수록 신초형성수 및 생장은 억제되었고 생체중 : 건물중의 비율도 감소하였다. 광도가 2000 lx일 때는 Bacto-agar 0.6-1.2%, 3,000 lx에서는 한천 0.4-0.6%에서 건전한 신초를 10개 이상 생산할 수 있었다. 활성탄소 0.1-0.3%첨가는 신초의 생장촉진과 뿌리형성에 촉진적으로 작용하였으며 투명화 발생률도 감소시켰으나 신초형성수는 현저히 감소시켰다. MS배지내 첨가되는 다량원소의 농도를 0.8-1.0배 하였을때 신초형성수 및 생장이 촉진되었다. 기관형성에 미치는 오옥신의 종류 및 농도별 효과를 조사한 결과 NAA나 IBA 보다는 IAA 0.3mg/L 와 BA 5.0mg/L를 혼합처리 하였을 때 증식률이 16배에 달하였다. Multiple shoots obtained in MS medium suppler with 5.0 mg/L BA though shoot-tip culture. The frequency of vitrified shoot was lower on Bacto-agar medium than on Gelrite as gelling agent. Addition of activated charcoal at concentrations of 0.1~0.3% reduced vitrification and markedly increased shoot growth, and formation and growth of roots, but significantly reduced the number of shoots formed. The ratio of fresh weight to dry weight was decreased by increasing light intensity and agar concentration. Eight-tenths times of macroelement of MS medium was observed to be effective for shoot formation. Addition of IAA effectively promoted shoot formation in both shoot tip and node-bud explants. Supplement of 5.0 mg/L BA, 0.3 mg/L IAA to MS medium was most effective in shoot proliferation on shoot tip and node-bud explants.Multiple shoots obtained in MS medium suppler with 5.0 mg/L BA though shoot-tip culture. The frequency of vitrified shoot was lower on Bacto-agar medium than on Gelrite as gelling agent. Addition of activated charcoal at concentrations of 0.1~0.3% reduced vitrification and markedly increased shoot growth, and formation and growth of roots, but significantly reduced the number of shoots formed. The ratio of fresh weight to dry weight was decreased by increasing light intensity and agar concentration. Eight-tenths times of macroelement of MS medium was observed to be effective for shoot formation. Addition of IAA effectively promoted shoot formation in both shoot tip and node-bud explants. Supplement of 5.0 mg/L BA, 0.3 mg/L IAA to MS medium was most effective in shoot proliferation on shoot tip and node-bud explants.
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백기엽,유광진,박상일 한국식물생명공학회 1997 식물생명공학회지 Vol.24 No.6
기내에서 증식된 다수의 지황(Rehmania glutinosa)신초를 가장 효율적으로 발근시키기 위해서 기내 및 기외발근 실험을 수행하였다. 오옥신의 종류와 농도를 달리하여 신초와 마디조직을 배양해본 결과 발근율은 거의 100%에 달하였으며 IAA 0.5~1.0 mg/L 처리구에서 절편체당 뿌리형성수가 가장 많았다. 배지내 당의 농도가 증가할수록 신초의 생장은 현저히 감소하였으나 발근율은 100%에 달하였고 형성된 뿌리의 굵기가 커지는 경향이였다. paclobutrazol과chloromequat를 처리해본 결과 마디나 신초조직에 관계없이 발근율은 거의 100%에 달하였으며 절편체당 근형성수도 억제제의 농도에 크게 영향을 받지 않았으며 신초조직에서는 5~7개, 마디조직에서는 3.5~5.0개가 형성되었다. 기외발근시에는 100ppm의 IBA용액에 15분~시간 침지처리하는 것이 타 농도보다 절편체당 근형성수가 증가하였다. 분말로된 발근촉진체를 이용한 경우 신초기부 절단면에 IBA 0.1%를 분말처리하여 버미큘라이트와 퍼얼라이트가 1:1로 혼합된 삽목상에 삽목하는 것이 생존율 및 발근율이 100%에 달하였다. 100% root formation in in vitro cultures was observed regardless of kind and levels of auxin used and explant source. The number of roots/explant was increased in 0.5~1.0 mg/L IAA treatment. Thicker roots were observed with the addition of 9% sucrose compared with medium containing lower sucrose concentrations. Paclobutrazol and chlormequat had no effect on tuberization of formed roots but slightly increased the number of root. In in vivo rooting, soaking of regenerated shoot cuttings to 100 mg/L IBA for 15 to 60 minutes was found effective. Treatment of 0.1% IBA rooting powder and planting in rooting medium composred of vermiculite(1) : perlite(1) gave 100% rooting and survival.
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난 자생지 토양이 조직배양한 춘란과 한란의 생장에 미치는 영향과 난 공생균의 관찰
김태중,이상선,박성수,백기엽 충북대학교 한국과학재단 지정 첨단원예기술개발연구센터 1996 연구보고서 Vol.1 No.-
현재, 난재배가들이 사용하는 다양한 용토와 춘란자생지 용토를 사용하여 춘란과 한란을 재배하였다. 용토 실험에서 자생지 토양과 바크가 춘란 재배에 가장 좋았으며, 한란재배에서도 비멸균한 토양에서 동일한 결과를 얻었다. 이러한 난생장과 관련되는 난뿌리에서는 난균근 형성과 관련되는 것으로 나타났다. 난균근이 형성되는 난뿌리 부분에서 질소 함량이 높게 측정되어, 이는 질소성분이 난생장에 효과적으로 작용하였다고 생각되었다. 이상의 결과를 근거로하여 난의 자생을 미친다는 것과 난균근의 차이점을 파악하였다. 또한 난균근 형성에서 난균근과 난의 상호관계에서 숙주와 균의 특이성을 관찰하였다. 본실험에 사용된 춘란자생지 토양에서는 최소한 동양란에 관련되는 공생균이 최소한 두 개 이상이라는 것을 밝혔으며, 이에 대한 공생과정도 서로 다를 것이라는 것으로 추측되었다. The temperate and terrestrial orchids of C. goeringii and C. Kanran were cultivated for 16 months under the conditions of green house, using the various culture media and the soils collected from Korean native orchid inhabiting areas (KNOA). The growth of C. goeringii were observed to grow well in the pine bark and the soil of KNOA as culture medium, and to form orchid mycorrhizae in the roots. The plantlets of C. kanran grew better in the non-sterilized soils than in the sterilized soils collected from two different KNOA. The values of total-nitrogen of the roots grown in the non-sterilized soils were measured to be higher than those in the other culture media, and even though higher than those of the shoots grown in the same soils. These growths of young plantlets of C. goeringii and C. karnan were revealed to be related to the orchid mycorrhizae, and speculated to be involved in the metabolism of nitrogen sources. Thus, based on this result, the soils of KNOA were collected from various areas, mainly Southern and Western Korea, and applied to cultivations of three different orient orchid plants. The formation of orchid mycorrhizae (mainly peletons in the root cells) and the periods of infection by orchid mycorrhizal fungi (8 to 12 weeks) were different for three different orchid plants. Host specificity were speculated to be resulted from the interactions between orchid plants and fungi. Also, more than two types of fungus were, at least, confirmed to be involved in the orchid mycorrhizae for the oriental orchids.
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