• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.10-10
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• 2023

투구꽃속(Aconitum L.)은 미나리아재비과(Ranunculaceae)의 가장 큰 속(Genus) 중 하나로 300-400 여종(Species)이 북반구 온대지역에 분포한다(Tamura, 1993). 둥근 투구모양의 꽃받침이 특징적이며, aconitine과 같은 알칼로이드 성분을 함유하고 있어 전통적으로 약용소재로서 활용되었다(Khorana, 1968). 세뿔투구꽃(Aconitum austrokoreense Koidz.)은 한국 경상도, 전라남도에만 국지적으로 자라는 한국 특산식물이자, IUCN에서 CR(Critical Endangered) 등급으로 지정된 희귀식물이다. 이 종은 다년생이지만, 서식지에서 개체수가 적거나 유묘의 발견이 어려운 등 자생지 축소 및 절멸의 우려가 높다. 세뿔투구꽃을 보호하고 종을 보존하기 위해서는 종자 발아 · 휴면에 대한 지속적 연구와 종자번식법의 개발이 필요하다. 본 연구에서는 세뿔투구꽃의 종자의 휴면유형을 분석하고 호르몬에 의해 휴면타파를 유도하였으며, 특히 발아 2단계에서 일어나는 생리적 변화를 단백질체학적 관점에서 분석하였다. 세뿔투구꽃 종자는 미발달된 배를 가지고 있으며, 충분한 수분과 온도의 조건에서도 발아가 관찰되지 않아 형태 · 생리적 휴면으로 판단되었다. 종자는 BAP 처리에서 휴면이 타파되는 것을 확인하였으며, 특히 BAP 농도에 따른 발아율은 통계적으로 유의한 값을 가졌다 (F=23.208, P<0.01). 건조된 종자에 비해 BAP 처리된 종자는 발아 2단계에서 몇가지 단백질의 증가가 관찰되었다. 대부분의 종자는 30S ribosomal protein subunit으로, 휴면상태에서 발아상태로 변환이 일루어지는 생리적 변화의 증거로 확인된다. 특히 발아2단계를 지나는 세뿔투구꽃 종자에서는 cytochrome subunit과 photosystem II protein의 급격한 증가가 관찰되었는데, 이는 배아의 발달 결과인 자엽이 발아 즉시 광합성이 가능하도록 준비하는 과정에서 발현 된 것으로 추정한다. 본 연구는 발아2단계에서 휴면타파에 직접적인 영향을 미치는 요인을 발견하지 못하였으나, 휴면타파에 중요한 단계인 발아 2단계 종자의 생리학적 변화를 이해하기 위한 기초자료로 활용 될 수 있을 것이다.

• Korean Journal of Plant Tissue Culture
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• v.28 no.1
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• pp.7-13
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• 2001

Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

• Environmental Mutagens and Carcinogens
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• v.21 no.2
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• pp.82-88
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• 2001

Recently, there has been significant progress in understanding the control process of the cell division cycle. To investigate the influence of toxic substances on the cell cycle, the effect of benzo(a)pyrene (BAP) and mitomycine C (MMC) on synchronized HeLa cells was analyzed during the cell cycle. To synchronize the HeLa cells, 10$^{6}$ cells were grown for 1 day and then treated with 1 mM hydroxyurea for 14 h. The arrested cells were then allowed to proceed through their cell cycle by removing the hydroxyurea and resupplying a fresh medium. The arrested cells in the G1/S transition then proceeded to the S phase after 4 h, the G2/M phase after 8h, and the G1 phase after 12 h, subsequent to the resupply of a fresh medium. In the untreated HeLa cells, the p34$^{cdc2}$ kinase activity, measured using a p34$^{cdc2}$ specific peptide, peaked after 8h (G2/M) and then declined after 12 h (G1). However, treatment with 30 $\mu$M BAP delayed the peak of the p34$^{cdc2}$ kinase activity. The amount of p34$^{cdc2}$ remained unchanged in the untreated, BAP-, and MMC-treated cells throughout the cell cycle. The cyclin B level peaked after 8 h in the untreated cells, yet peaked after 10-12 h in the BAP-treated cells. There was no significant change in the cyclin B level in the MMC-treated cells.

• Korean Journal of Medicinal Crop Science
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• v.12 no.6
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• pp.500-507
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• 2004

The establishment of an efficient protocol of plant regeneration from leaf explant cultures of Perilla spp. is reported. Regenerated shoots were obtained from leaf explant cultures on solid MS medium containing different concentrations of cytokinins and auxin. The effect of cytokonin and auxin differed depending on each acession. The combination treatments of high level of cytokinin and low level of auxin was more effective for plant regeneration in Perilla frutescens. The best concentration of sucrose was 3% for regeneration. Of spermidine, spermin and putrescine. treatments, the most effective treatment for plant regeneration was $10\;mg/{\ell}$ spermidine.

• Korean Journal of Agricultural Science
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• v.14 no.2
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• pp.205-212
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• 1987

To examine the morphogenetic response, stem segments of 8 Populus spp. and 3 different explants of P. nigra var. italica were cultured on MS (Murashige and Skoog 1962) and WPM (Woody Plant Medium) medium containing various phytohormones. The results obtained were as follows: 1. Shoot regeneration and development from stem segment of 8 Populus spp. showed a quite difference according to the section and the species. All of the species of Leuce and Tacamahaca section did not form adventitious buds, while most of explants showed axillary or dormant bud elongation after 4 weeks. But P. nigra var. italica of Aigeiros section showed a successful adventitious bud formation (mean 5.4 buds per explant). 2. Leaf, petiole, and internode segment of P. nigra var. italica showed a quite differences according to media and ex plants upon the morphogenetic response. Adventitious bud formation from leaf was more abundant and readily initiated on the abaxial side than on the adaxial side. Mean number of 103 adventitious buds per explant was obtained from abaxial side of leaf segment cultured on WPM medium containing $0.2mg/{\ell}$ BAP for 5 weeks. 3. 2,4-D (2,4-dichlorophenoxy acetic acid) supplemented to media appeared to be negative upon the adventitious bud formation of P. nigra var. italica, while it promoted callus formation from all explants. Especially, NAA (${\alpha}$-naphtalene acetic acid) or NAA combination with BAP (6-benzylaminopurine) promoted root regeneration from the all explant of P. nigra var. italica in this study.

• Journal of Korean Society of Forest Science
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• v.77 no.2
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• pp.208-215
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• 1988

The leaf mesophyll protoplasts of Populus alba ${\times}$ glandulosa were isolated from leaf of plantlet in vitro and cultured for plant regeneration. The MS medium (minus $NH_4NO_3$) with 0.5 mg/l BAP and 2.0 mg/l 2, 4-D showed the moderate frequency of dividing protoplasts cultured by the liquid plating method during the first week of culture. The percentage of colony formation was revealed the highest frequency by the gauze contained semi-solid agar plating method after 5 weeks cultured. Ridding out the gauze, the micro-callus was formed on the same semi-solid medium in 8 weeks after protoplasts culture. For proliferation of callus, mini-callus was transferred on the MS solid medium with 0.5 mg/l 2, 4-D and 0.1 mg/l BAP 12 weeks after culture. Shoot regeneration occurred when the calli derived from protoplasts were cultured on MS medium with 1.0 mg/l zeatin and such shoots could be readily rooted on the one half strengthen MS medium with non-phytohormone. Rooting shoots were planted in green-house 22 weeks after protoplast culture.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.111-115
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• 2002

Taraxacum platycarpum has been used as a medicinal plant. We investigated optimal condition for efficient plant regeneration through adventitious shoot formation on medium with various kinds of growth regulators. Adventitious shoot formation was achieved when cytokinin was used alone. Shoot formation was higher on MS medium containing 2 mg/L BAP compared to that with 2 mg/L kinetin and 2 mg/L 2-ip. Among root, hypocotyl and cotyledon, roots were the best explant for the adventitious shoot induction. Adventitious shoot formation from roots declined markedly by the combination of both 0.1 mg/L NAA and 2 mg/L BAP, while shoot formation from cotyledons was stimulated by the same combination. Root formation from the regenerated shoots was achieved on 1/3MS medium containing 0.2 mg/L NAA. Regenerated plantlets was acclimatized and transplanted to the soil, showing 100% survival.

• Applied Biological Chemistry
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• v.35 no.2
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• pp.99-105
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• 1992

Artemisia annua contains the antimalarial principle, artemisinin. The possibility of the production of this compound through tissue culture technique was studied. The optimum combinations of hormones for the induction of callus were p-chlorophenoxyacetic acid(pcPA) and 6-benzylaminopurine(BAP) or pcPA and N-isopentenylaminopurine(2iP) in 0.05 mg/l each. For the growth of callus, the same combination of pcPA and BAP was optimum in concentrations of $1.0\;{\mu}M\;and\;0.5\;{\mu}M$, respectively, and the optimal concentration of sucrose was also found to be 2%(w/v). Tissue culture from the crown gall grew faster than normal callus. In the suspension culture broth and the cells of normal callus or Agrobacterium-transformed tumors, arteannuic acid and 11,12-dihydroarteannuic acid were found together with common phytosterols, whereas artemisinin was not found.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.195-200
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• 1998

Callus induction from leaf and stem explants of Arabidopsis thaliana was successfully obtained when leaf explants were cultured on MS medium containing 2.0 mg/L 2,4-D in the dark and also, when stem explants were cultured on CP medium containing 0.5 mg/L 2,4-D and 0.1 mg/L BAP. Explant-derived sliced calli were suspension-subcultured every week in CP liquid medium with 0.5 mg/L 2,4-D and 0.1 mg/L BAP in the dark, and shoot-forming cell clusters of nodular, pale yellow and knobby type were selected after 7-8 weeks of culture. Shoots were initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium containing 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for four weeks. Shoot regeneration frequency (calli regenerating at least one shoot) was more than 50%. For plant regeneration, excised shoots were trnasferred to hormone free medium for root initiation after 4 weeks of culture. The regenerants were bolting after 2 weeks of culture and formed in vitro flowering buds within bracts after 4 weeks of culture.

• Korean Journal of Plant Tissue Culture
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• v.28 no.2
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• pp.69-74
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• 2001

Calli were induced on MS medium supplemented with 0.5 mg/L 2,4-D by using the leaf explants of haploid which were derived from the diploid and haploid of Nicotiana tabacum cv BY4. These calli were subcultured on MS medium with the combination of 2.0 mg/L 2,4-D, 1.0 mg/L kinetin and 0.1 mg/L BAP. Cell propagation of diploid plants were good in a combination of 2.0 mg/L 2,4-D, 0.1mg/L BAP in vitro conditions, suspension cultures were conducted in equal condition. Homogenized suspension cultured cells were smeared 2.0 mL each on MS medium with 0~100 $\mu$M PFP, to select the resistant colony to PFP, and were examined after 10d, 20d and 30d. Measurment of fresh weight of cells after 30d of culture shows that with more concentration of PFP in medium the fresh weight of the cells decreased. In case of diploid, selected callus was the highest in vitro treated with 5 $\mu$M PFP. It was higher than control until 100 $\mu$M PFP. The active degree of catalase was the highest in vitro with 5 $\mu$M PFP but the lowest in vitro with 10 $\mu$M PFP on the other hand, in case of haploid plant, the active degree of peroxidase and catalase was the highest in vitro treated with 50 $\mu$M PFP. It's sure that enzyme active degree of between diploid and haploid had big differences.