• 한국산림과학회지
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• 제96권2호
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• pp.178-182
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• 2007

Rauvolfia serpentina (L.) Benth. ex Kurz is a medicinal plant and an endangered tropical rainforest plant species. Since the field cultivation that aims to fulfill the industrial needs is never accomplished, tissue culture appears to be the most feasible way to improve the quality and quantity of R. serpentina. This experiment used two kinds of explants (roots and shoots) to induce optimal root formation in different combinations of auxin and photoperiod. Each explants exhibited different responses on given treatments. Differentiated root could be produced from explants cultured in IBA 20 mg/L with and without light. The highest number of roots, root length and root weight induced from shoot explants were effective on MS medium containing IBA 20 mg/L and incubated under dark condition, while highest total weight (callus and root) from root explants cultured on MS medium supplemented 10 mg/L IBA and 10 mg/L NAA and incubated under day length (11/13 hr). The root induced from shoot explants produced the highest major alkaloid content. The highest content of ajmaline (2.17 ppm fresh weight) and reserpine (1.30 ppm fresh weight) were observed in shoot explants cultured in MS medium containing combination of IBA 10 mg/L and NAA 10 mg/L and incubated under dark condition, yohimbine (1.47 ppm fresh weight) was in the shoot explants cultured in MS medium containing NAA 20 mg/L and incubated under day length, while serpentine was absent.

• Journal of Plant Biotechnology
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• 제36권3호
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• pp.236-243
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• 2009

The halophyte Leymus chinensis (Trin.) is a perennial rhizome grass (tribe Gramineae) that is widely distributed throughout China, Mongolia and Siberia. This study was conducted to investigate an optimal condition for plant regeneration from mature seeds, leaf base segments, and root segments in L. chinensis. Plant growth regulators affecting embryogenic callus induction and plant regeneration were investigated by four-factor-three-level [L9 (34)] orthogonal test in this study. The effects of explants types (mature seeds, leaf base segments and root segments), callus types, medium types were examined in this study. Wild type (WT) and Jisheng No. 1 plants (JS) were used for primary callus induction. A clear explants difference was seen during callus induction; mature seeds were considered as the preferred explants; and the highest frequency of callus induction was obtained in Medium 6 using mature seeds as explants in WT. Plant regeneration ability was evaluated by frequencies of green callus forming, shooting, rooting, and shooting with roots. Effect of α-naphthalene acetic acid (NAA) on shoot regeneration was remarkable with the highest frequency of 70.8% in WT after 2-month culture. The medium with 0.2- 0.5 mg/L NAA was found to have the highest shoot induction. All regenerated shoots were successfully rooted when transferred on half-strength Murashige and Skoog (MS) basal medium. The acclimatized plantlets were grown to mature with flowering and seeds setting in green house conditions.

• Journal of Plant Biotechnology
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• 제36권1호
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• pp.81-86
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• 2009

An efficient and reproducible plant regeneration system was established using transverse thin cell layers (tTCLs) in five cultivars of Brassjca juncea L. The effects of medium conditions, explant types (tTCLs of hypcotyl and cotyledonary petiole) on shoot regeneration were examined in this study. The maximum shoot regeneration frequency was obtained in Murashige and Skoog (MS) medium supplemented with 4 mg/L 6-benzylaminopurine (BA) and 0.2 mg/L 1-naphthaleneacetic acid (NAA). The hypocotyls derived tTCL explants had more shoot regeneration frequency (52%) than the cotyledonary petiole derived tTCL explants. Shoot induction was further improved by the addition of silver nitrate ($AgNO_3$) in the regeneration medium. A significant genotypic effect was also observed between the five cultivars; Rai-5 displayed higher capacities to produce shoots than other cultivars. Regenerated shoots were rooted on MS basal medium without PGRs which induced 90% of roots. The plantlets established in greenhouse conditions with 99% survival, flowered normally and set seeds. The regenerated plants were fertile and identical to source plants.

• Journal of Plant Biotechnology
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• 제30권3호
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• pp.241-244
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• 2003

Root cluster section culture, showing high efficient Protocorm-like body (PLB) formation capacity, were established in Doritaenopsis hybrids. Three types of root were obtained from excised shoots in 1/2MS medium containing different concentrations of NAA; \circled1normal roots, \circled2multiple roots and \circled3abnormal root clusters. Those were placed on 1/2MS medium supplemented with 0.5 mg/L thidiazuron for PLB regeneration. PLB regeneration rate was greater in root cluster section cultures (77.8%) compare to normal root tip cultures(30%). Number of PLBs regenerated from root cluster sections were counted over 11 per explant (5.3 per normal root tip).High frequency of PLB regeneration was achieved in root cluster section culture. This result can be used as an efficient method for clonal proliferation of Doritaenopsis hybrids.

• Journal of Plant Biotechnology
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• 제37권2호
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• pp.228-235
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• 2010

The establishment of cell suspension culture and plant regeneration of the halophytic Leymus chinensis (Trin.) are described in this study for the first time. Callus induction solid medium containing Murashige and Shoog (MS) basic salt, $2.0\;mg\;l^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), and $5.0\;mg\;l^{-1}$ L-glutamic acid with $30.0\;g\;l^{-1}$ sucrose and $4.0\;g\;l^{-1}$ gelrite for solidification induced the highest rate of cell division in Type 1 callus among calli of various types. Liquid medium with the same hormone distribution was therefore, used for cell suspension culture from Type 1 callus. Over a 30 d suspension culture at 100 rpm, great amounts of biomass were accumulated, with 71.07% average daily increment and 22.32-fold total fresh weight increment. Comparison of before and after suspension culture, the distribution of different size callus pieces and the maintenance of callus type were basically unaltered, but a slight increase in relative water contents was observed. To induce the potential of plant regeneration, the directly transferring on plant regeneration solid medium containing MS basic salt, $0.2\;mg\;l^{-1}$ $\alpha$-naphthalene acetic acid (NAA), $2.0\;mg\;l^{-1}$ kinetin (Kn), and $2.0\;g\;l^{-1}$ casamino acid and indirectly transferring were simultaneously performed. Even now growth rates of suspension-derived callus on solid medium were approximately half of those of Type 1 callus, but faster somatic embryogenesis was observed. Rooting of all regenerated shoots was successfully performed on half-strength MS medium. All plants appeared phenotypically normal.

• Journal of Plant Biotechnology
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• 제44권1호
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• pp.42-48
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• 2017

본 연구는 윤판나물의 효과적인 기내 증식을 위한 배양 최적 조건을 구명하기 위해 수행되었다. 신초 유도 및 생장에 영향하는 배지(MS, B5 및 WPM) 및 절편체 종류에 따른 실험에서 줄기 유도는 액아 절편을 식물생장조절물질 무첨가 MS 배지에 배양 시 절편 당 2.5개로 가장 좋았고, 줄기 신장은 정아 절편을 식물생장조절물질 무첨가 MS 배지 배양 시 7.2 cm로 가장 효과적이었다. BA 침지 처리 시 신초 유도 효과는 액아배양 유래 신초를 BA 침지 무처리구에서 신초 유도(2.29개/절편) 및 신초 신장(7.28 cm)이 가장 효과적이었으며, BA 침지 시간이 길어짐에 따라 신초 유도 및 길이 생장은 감소하였다. 발근에 영향하는 IBA 농도 효과는 정아유래 신초를 1.0 mg/L 처리구에서 배양 시 최대 발근율(100%) 및 뿌리 유도수(21.3개/절편)를 보였으며, 정아 혹은 액아 유래 신초지 종류에 따른 발근율의 차이는 크지 않은 것으로 관찰되었다. 발근된 식물체는 상토로 이식하여 형태적인 변이 없이 정상적인 생장을 보였다.

• 식물조직배양학회지
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• 제28권2호
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• pp.61-67
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• 2001

들잔디 (Zoysia japonica Steud.)의 성숙종자 유래의 캘러스를 사용하여 캘러스 생장 및 재분화에 적합한 몇 가지 조건과 캘러스 유형에 따른 재분화 효율을 검토하였다. 종자로부터 캘러스 유도는 2 mg/L 2,4-D, 0.2 mg/L BAP, 4 mg/L thiamine-HCl, 100 mg/L $\alpha$-ketoglutaric acid를 포함한 MS 배지에서 가장 좋았다. 캘러스 증식은 0.5 mg/L 2,4-D, 0.05mg/L BAP, 4 mg/L thiamine-HCl, 100 mg/L $\alpha$-ketoglutaric acid를 포함한 MS 배지가 가장 좋았다. 식물체의 재분화는 3% maltose, 1 mg/L BAP 또는 1 mg/L TDZ를 포함한 MS배지에서 좋았다. 캘러스의 Type과 빈도는 2,4-D와 BAP 조합과 농도에 따라 크게 영향을 받았고, 형태적으로 4가지 Type이 뚜렷하게 나타났다. Type I, II, III 캘러스는 계대배양으로 신초가 형성된 반면, 물기가 많은 캘러스인 Type IV에서는 뿌리만이 분화되었다. 이 중 Type I 캘러스는 신초의 재분화율이 82%로 가장 높고 albino체의 출현빈도가 4%로 가장 낮았다. 이와같이 캘러스를 유형별로 구분하여 배양함으로써 잔디 캘러스로부터 식물체의 재분화를 높일 수 있었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2021년도 춘계학술대회
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• pp.36-36
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• 2021

Cryopreservation has been broadly used as an efficient method for a long-term conservation for many types of plants especially vegetatively propagated plants. Among several cryopreservation methods, a droplet-vitrification was the most widely applicable and efficient method. Studies have developed protocols for strawberry using droplet-vitrification method and suggested the practical use of the protocol for large number of germplasm with a little modification. In this study, the droplet vitrification method of shoot tip has been tested on 31 accessions provided around the world. Shoot tips were precultured on Murashige and Skoog (MS) liquid medium supplemented with 0.3~0.5M sucrose. Precultured explants were osmoprotected with loading solution, 35% of PVS3 (C4, 17.5% glycerol and 17.5% sucrose) for 40 min and exposed to dehydration solution, PVS3 (B1, 50% glycerol and 50% sucrose) for 60 min. Then, the explants were transferred onto droplets containing 2.5 uL PVS3 on sterilized aluminum foils prior to direct immersion in liquid nitrogen (LN) for 1hr. The cryopreserved shoot tips were rapidly warmed in a water bath at 40C and then unloaded in MS with 0.8M sucrose for 40 min. The shoot tips were cultured in NH₄NO₃-free MS post culture medium for 2 weeks. Subsequently, the explants were moved to the MS medium for 6 weeks and evaluated the regrowth rate. By this droplet-vitrification protocol, twenty-four accessions showed at least 40% regrowth rate. Out of 24 accessions, 'Nonsan1ho' had the highest regeneration rate of 85.8% and 'Jumbo pureberry' had the lowest with 42.1%.

• 식물조직배양학회지
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• 제27권4호
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• pp.309-315
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• 2000

Panax ginseng is an important medicinal plant that has been used worldwide for geriatric, tonic, stomachic, and aphrodisiac treatments. Ginsenosides contained in the ginseng root are the main substances having active functions for human body. The price of ginseng is very expensive due to a complex process of cultivation, and the yield of ginseng is limited, which cannot meet the demand of the increasing market. Researchers have applied plant biotechnology to solve the problems but there are still things to be determined towards ginsenoside production by large-scale adventitious root culture. In this experiment, 5 to 20 liter bioreactors were employed to determine optimal conditions for adventitious root culture and ginsenoside production of Panax gineng. Callus was induced from the ginseng root on MS agar medium containing 1.0 mg. $L^{-1}$ 2,4-D and 0.1 mg. $L^{-1}$ kinetin. Then the callus was cultured on MS agar medium supplemented with 2.0 mg. $L^{-1}$ IBA, 0.1 mg. $L^{-1}$ kinetin, and 30 g. $L^{-1}$ to induce adventitious roots. The maximum root growth and ginsenoside production were obtained in 1/2 MS medium. 2.0 mg. $L^{-1}$ naphthalene acetic acid resulted in greater root growth than 2.0 mg $L^{-1}$ indole-3-butyric acid. Ginsenoside content increased with 2.0 mg. $L^{-1}$ benzyl adenin or kinetin. High concentrations of benzyl adenin (above 3.0 mg. $L^{-1}$ ) decreased the adventitious root growth and ginsenoside productivity. N $H_{4}$$^{+}$ inhibited the ginsenoside accumulation, while high concentrations of $K^{+}$, $Mg_{2}$$^{+}$, and $Ca_{2}$$^{+}$ increased it. N $H_{4}$$^{+}$ at 0.5 and 1.0 times of the normal amount in 3/4 SH medium resulted in the greatest biomass increase, but the highest ginsenoside productivity was obtained when N $O_{3}$$^{-}$ was used as the sole nitrogen source in the medium. Most microelements at high concentrations in the medium inhibited the root growth, but high concentrations of MnS $O_4$enhanced the root growth. Root dry weight increased with increasing sucrose concentrations up to 50 g. $L^{-1}$ , but decreased from 70 g $L^{-1}$ Ginsenoside productivity was maximized at the range of 20 to 30 g. $L^{-1}$ sucrose. In the experiment on bioreactor types, cone and balloon types were determined to be favorable for both adventitious root growth and ginsenoside production. Jasmonic acid was effective for increasing ginsenoside contents and Rb group ginsenosides mainly increased. These results could be employed in commercial scale bioreactor cultures of Panax ginseng.x ginseng.

• Journal of Plant Biotechnology
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• 제30권3호
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• pp.233-239
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• 2003

We have developed a reliable and high-frequency genetic transformation and regeneration system via somatic embryogensis of Eleutherococcus sessiliflorus. Embryogenic callus obtained from seed were co- cultivated with Agrobacterium tumefaciens strain EHA101/pIG121Hm harboring genes for intron-$\beta$-glucoronidase(GUS), kanamycin and hygromycin resistance. Following co-cultivation, two types of samples(fine embrogenic calli and early globular embryo clusters) were cultivated on Murashige and Skoog(MS) medium containing 1 mg/L2.4-D for 3day in dark. Transient expression of GUS gene was found to be higher in the early globular embryo clusters than in the embryogenic calli. Also, co-cultivated period affected expression of GUS gene; the best result was obtained when globular embryo clusters were co-cultivated with Agrobacterium for 3 days. Subsequently, this callus transferred to selective MS medium containing 1mg/L2.4-D, 50mg/L kanamycin or/and 30mg/L hygromycin and 300mg/L cefortaxime. These embryogenic calls were subcultured to the same selection medium at every 2 weeks intervals. Approximately 24.5% of the early globular embryos co-cultivated with Agrobacterium for 3days produced kanamycin or/and hygromycin-resistant calli. Transgenic somatic embryos were converted into plantlets in half strength MS medium supplemented with 3mg/L GA$_3$ kanamycin and were confirmed by GUS histochemical assay and polymerase chain reaction analysis. Genomic Southem blot hybridization confirmed the incorporation of NPT II gene into the host genome.