• 한국약용작물학회지
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• 제19권1호
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• pp.9-15
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• 2011

This study was conducted to establish the optimal condition for conservation of genetic resources and micropropagation of Lilium cernum. Induction of bulbet of L. cernum was highly effective (9.2 bulb/explant) on 1/2 SH (Schenk and Hildebrandt) medium supplemented with 1.0 mg/L TDZ (Thidiazuron) and 0.1 mg/L NAA (Naphthaleneacetic acid). The treatment of 0.1 mg/L NAA increased root development (6.4 root/explant) under the in vitro condition. In addition, treatments of AC (Activated Charcoal) and ventilation were enhanced to develop number of shoots and to elongate length of leaf, bulb and root. Futhermore, the process of short-term soil acclimatization was promoted to strengthen the plantlets induced under the in vitro condition.

• 한국자원식물학회지
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• 제28권6호
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• pp.690-696
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• 2015

We micropropagated pear (Pyrus species) using shoot tips and nodal explants from three pear genotypes. The ability to establish shoot tip cultures, proliferate shoots, induce rooting, and acclimatize the resulting plantlets are all elements of in vitro micropropagation. Shoots were induced from shoot tips on Murashige and Skoog medium (MS) with five different plant growth regulator combinations. The highest shoot formation rates were achieved for the three genotypes using MS supplemented with 1.0 mg/L N⁶-benzyladenine (BA) and 0.1 mg/L gibberellic acid (GA₃). The maximum shoot number and shoot length for the three cultivars were recorded with 2.0 mg/L BA and 0.2 mg/L indole-3-butyric acid (IBA) in multiplication medium using nodal explants produced from microshoots. Nodal explants with one or two axillary buds cultured for three weeks initiated roots on medium supplemented with various concentrations of 1-naphthaleneacetic acid (NAA) or/and IBA in half-strength MS medium for adventitious rooting. The highest rooting response was with the combination of 0.2 mg/L NAA and 0.2 mg/L IBA. A combination of NAA and IBA resulted in a significant increase in the rooting ratio over NAA or IBA alone. In this medium, the root formation rate according to ranged from 68.9% for the BaeYun No. 3 genotype to 51.8% for the Hwanggeum genotype. We also investigated the influence of the concentration the polyamine phloroglucinol in rooting medium. For all three genotypes, the highest rooting ratio, longest root length, and greatest root number were observed in the treatments with 75-150 mg/L phloroglucinol. Most rooted plants were acclimatized successfully.

• 한국식품영양과학회지
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• 제17권2호
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• pp.115-124
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• 1988

비타민C의 급원식품인 콩나물의 생장과 비타민C의 함량을 식물 생장조절제에 의하여 조절할 목적으로 gibberellic acid$(GA_3)$, indole-3-acetic acid(IAA) 및 1-naphthaleneacetic acid(NAa)의 단독 및 혼합 처리를 하였을때 가장 적당한 식물 생장조절제의 종류 및 그 농도를 찾고 이 생장조절제가 비타민C의 생합성에 미치는 영향 및 당대사와 비타민C 생성과의 관계를 검토한 결과를 요약하면 다음과 같다. 콩나물의 생장은 NAA는 $10^{-8}M$, IAA 는 $10^{-6}M$, $GA_3$는 $10^{-5}M$에서 각각 가장 양호하였다. $GA_3$와 NAA 혼합처리의 경우는 $GA_3\;10^{-5}M+NAA\;10^{-9}M$에서, $GA_3$와 IAA 혼합처리의 경우는 $GA_3,\;10^{-5}M+IAA\;$10^{-9}M$에서 양호하였고, 단독처리가 혼합처리에 비하여 양호하였으며 특히 IAA $10^{-6}M$ 처리에서 가장 좋았다. 생장이 비교적 양호한 NAA $10^{-8}M, IAA\;10^{-6}M,\;GA_3\;10^{-5}M,\;GA_3\;10^{-8}M+IAA\;10^{-6}M$ 및 $GA_3\;10^{-5}M+IAA\;10^{-9}M$의 각 처리구에서 비타민C의 생성 정도를 조사한 결과 콩나물 생장이 가장 양호하였던 IAA $10^{-6}M$ 에서 24.26mg%로서 그 함량이 가장 높았으며 대조구의 1.6배였다. 생장이 저해되지 않는 농도의 chloramphenicol(CAP) 처리에 의하여 ribulose diphosphate carboxylase(RuDpCO)는 저해되었으나 nicotineamide adenine dinucleotide phosphate-glyceraldhyde-3-phosphate dehydrogenase (NADP-GDH), galactonolactone dehydrogenase (GLD)의 활성 및 비타민C 함량은 저해되지 않았다. 따라서 비타민C의 생합성은 chloroplast RNA의 작용과는 직접적인 관련은 없고 cytoplasm과 밀접한 관련이 있다고 생각되었다. $IAA\;10^{-6}M$ 처리에 의하여 GLD 활성은 대조구의 1.8배로 촉진되었다. IAA $10^{-6}M$은 chloroplast와 cytoplasm RNA와 단백질의 생합성을 크게 촉진하였으며 cytoplasm에서 더욱 활발하였다. 따라서 IAA는 cytoplasm RNA를 조절하므로 여기서 지배되는 GLD의 생합성을 촉진시킨 결과 비타민C의 생합성이 증가된 것으로 생각되었다. 산화적 인산화 반응의 짝풀림 물질이 2,4+dinitrophenol 처리로 비타민C의 생합성은 저해되지 않았으나, 해당 저해제인 monoiodoacetate와 NaF 및 TCA 회로 저해제인 $NaN_3$와 KCN은 다같이 콩나물의 생장과 비타민C의 생합성을 크게 저해하였다

• Plant Biotechnology Reports
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• 제4권2호
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• pp.173-178
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• 2010

An efficient protocol for shoot regeneration was developed for sesame (Sesamum indicum L.) internodes using the transverse thin cell layer (tTCL) culture method. The frequency of shoot regeneration and the number of adventitious buds produced from regenerated shoots depend significantly on explant age, thickness of the tTCL sections, and the phytohormones supplemented to the culture medium. A combination of 6-benzyladenine (2.0 $mg\;l^{-1}$) and a-naphthaleneacetic acid (0.5 $mg\;l^{-1}$) was found to be the best phytohormone combination for shoot bud induction, with the maximum number of shoots obtained when the tTCL sections were 0.5-1.0 mm thick and derived from 4- to 6-week-old seedlings of sesame. Well-developed shoots were rooted on MS medium without phytohormones, and 80% of the regenerated plantlets were successfully established in soil.

• 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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• 제45권4호
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• pp.382-391
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• 2018

본 연구는 사과 대목 M.26 (Malus pumila Mill))의 효과적인 기내 대량번식하기 위해 신초 증식과 뿌리 형성에 적합한 배지조건을 확립하고, simple sequence repeat (SSR) 마커를 이용하여 증식된 소식물체의 유전적 다양성을 분석하고자 수행하였다. MS (Murashige and Skoog) 기본배지에 benzyladenin (BA, $0.5{\sim}5.0mg{\cdot}L^{-1}$)와 thidiazuron (TDZ, $0.01{\sim}0.1mg{\cdot}L^{-1}$)을 첨가하여 신초를 배양한 결과, $1.0mg{\cdot}L^{-1}$ BA 처리에서 절편체 당신초 수가 10.67개로 가장 많았으며 과수화 발생률은 BA 처리구보다 TDZ 처리구에서 높았다. M.26 신초 증식에 BA와 auxin과의 혼용처리 효과는 없었고, $1.0mg{\cdot}L^{-1}$ BA가 첨가된 MS 기본배지가 적합하였다. 신초 발근에 적합한 배지를 구명하고자 auxin인 indole-3-butyric acid (IBA)와 ${\alpha}$-naphthaleneacetic acid의 농도($0.5{\sim}5.0mg{\cdot}L^{-1}$), MS 배지의 무기염류(1/4 ~ 1배) 및 sucrose 농도($0{\sim}30g{\cdot}L^{-1}$)를 달리하여 처리한 결과, $1.0mg{\cdot}L^{-1}$ IBA, $15{\sim}20g{\cdot}L^{-1}$ sucrose가 첨가된 1/2 MS 배지에서 발근율(100%), 뿌리 수(10.45 ~ 13.60개/절편체), 뿌리 길이(7.41 ~ 8.33 cm) 및 신초 길이(4.93 ~ 5.38 cm)가 양호하였다. 15종의 SSR primer를 이용하여 증식된 20개의 소식물체를 분석한 결과, 총 30개의 대립유전자가 검출되었으며 모두 동일한 밴드 패턴을 보여 온실에서 자란 M.26 식물체와 유사하여 유전적으로 안정한 것으로 판단되었다.

• Journal of Plant Biotechnology
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• 제45권4호
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• pp.347-356
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• 2018

Chionanthus retusus is a small deciduous tree that is widely used in landscaping due to its beautiful white spring flowers and ornamental value. Conventional propagation through seeds requires one to two years of breaking dormancy. The objective of this study was to determine the conditions of in vitro germination in C. retusus. In vitro embryo culture was carried out to investigate the effects of six factors: basal media (McCown Woody Plant Medium (WPM) and Murashige and Skoog (MS)); plant growth regulators (different combinations and concentrations of naphthaleneacetic acid (NAA), 6-Benzylaminopurine (BA), and gibberellic acid ($GA_3$)); embryo age (collected weekly beginning 36 days after fruit setting); low temperature pretreatment (storing $4^{\circ}C$ for 1, 2, 3, and 4 weeks); coconut additives (100, 200, and $300ml{\cdot}L^{-1}$); and genotype (grouping plants depending on their flowering nature). The basal medium used in this study was WPM with $2mg{\cdot}L^{-1-1}\;GA_3$, $20g{\cdot}L^{-1}$ sucrose, and $6g{\cdot}L^{-1}$ Agar. WPM medium mixed with $GA_3$, resulted in higher germination rate as compared to when using a combination of auxin and cytokinin. $GA_3$ at $2mg{\cdot}L^{-1}$ was the most effective of all combinations and concentrations of PGRs. WPM medium with $2mg{\cdot}L^{-1}GA_3$ resulted in better and faster germination (75.93%). Embryos collected at 57 days after fruit setting had the highest percent of germinated seeds (87.04%) while low-temperature pretreatment of fruits at $4^{\circ}C$ for two weeks produced the highest germination (95.37%). These results of this study could be an open ground for development of an efficient protocol for commercial production of the ornamental tree.

• Journal of Plant Biotechnology
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• 제48권1호
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• pp.54-61
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• 2021

This work describe an efficient method for the shoot induction and plant regeneration of seedling-derived apical bud explants of Tilia mandshurica Rupr. & Maxim. The highest rate of shoot induction (82.2%) was obtained when apical bud explants from juvenile seedlings (5 months old) were cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BAP). However, apical bud explants obtained from mature trees (12 years old) did not produce any shoots, even with BAP supplementation. Among the three cytokinins tested for shoot multiplication (BAP, zeatin, and kinetin), BAP was the most effective; the highest number of shoots per explant (2.1) was observed on MS medium supplemented with 1.0 mg/L BAP. In contrast, the longest average shoot length (3.0 cm) was observed after growth on MS medium with 2.0 mg/L zeatin. No multiplication occurred when apical bud explants were cultured with kinetin-supplemented media. During rooting of in vitro-elongated shoots, the highest rooting rate (100%) was observed in half-strength MS medium supplemented with 0.5 ~ 1.0 mg/L indole-3-butyric acid (IBA) or 3.0 mg/L 1-naphthaleneacetic acid (NAA). During the acclimatization process, plantlets that were rooted on the IBA (0.5 mg/L)-supplemented medium had the highest survival rate (100%) and maximum root length (18.5 cm). These findings suggest that a low concentration (0.5 mg/L) of IBA is appropriate for the rooting and acclimatization of T. mandshurica. Plants were successfully transferred to the greenhouse with a 100% survival rate. This protocol will be useful for the large-scale propagation of Tilia species.

• Journal of Plant Biotechnology
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• 제46권2호
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• pp.114-118
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• 2019

Efficient protocol for plant shoot regeneration of Brassica juncea L. CZERN was established by using organic media components and growth stimulating factors of the vermicompost and coelomic fluids. Formulated organic plant tissue culture media (Vermicompost (30%) extracts supplemented with 20 mL/L coelomic fluid) have shown maximum shoot regeneration when compared with the Murashige and Skoog (MS) medium, which were supplemented with 1 mg/L 6-benzyladenine (BA) and 0.1 mg/L of Naphthaleneacetic acid (NAA). Cotyledon explants produced the highest shoot regeneration frequency from fourday-old germinated seedlings in comparison with non-germinated seedlings. The vermicompost extracts have proved to be the best organic plant growth media to induce shoots from cotyledons compared to the MS media. Statistically significant difference (P = 0.008) for the root length, shoot length (P=0.000350) and the leaves (P=0.375) of the mustard plantlets were analyzed successfully. The survival rate was 98% in the mustard cotyledons on the Vermicompost extract media and 63% on MS media respectively. The coelomic fluid also is much suitable to induce shoots from cotyledons at lower concentrations. It was also shown that the vermicompost extract, which comprised of humic acids along with coelomic fluid, affected shoot regeneration from the cotyledons. An efficient and organic shoot regeneration study was standardized and it can be applicable in the improvement of the economically important crops.

• Journal of Plant Biotechnology
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• 제34권4호
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• pp.293-298
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• 2007

Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.