• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.227-232
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• 2000

This study was carried out to elucidate the effect of cadmium on somatic embryogenesis and plant regeneration from cultured cells of Daucus carota L. Embryogenic calli were induced from cotyledon explants of carrot seedlings cultured on MS solid medium supplemente with 1 mg/L 2,4-D Embryogenic cells proliferated on medium supplemented with 1 mg/L 2,4-D were also cultured in liquid MS medium containing various concentrations (50, 100, 200, 500, 1000 $\mu$M) of cadmium for one week and then transferred to MS basal medium. Somatic embryogenesis occurred in suspension culture treated with 50 $\mu$M and 100 $\mu$M cadmium or untreated with cadmium. When cadmium was treated in suspension culture, production of two and four cotyledonary somatic embryos was reduced, but that of three cotyledonary somatic embryo was increased. Two cotyledonary embryos showed higher regeneration frequency than abnormal somatic embryo with one, three and four cotyledon. Regardless of cotyledonary variation, germination frequency of somatic embryos treated with cadmium was decreased in compared with that of embryos in basal medium.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.121-130
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• 2017

This study was carried out to investigate a method for producing cultured virus - free ovules for breeding high - quality Citrus cultivars. Ovules from the immature fruits of three citrus cultivars native to Jeju (Dongjeongkyool, Cheongkyool, and Jikak) and two cultivars of Citrus unshiu Marc. (Miyagawa wase and Haryejosaeng) that were thought to be infected with Citrus tristeza virus (CTV) were cultured on MS2 medium (Murashige - Skoog [MS] basal medium containing $500mg{\cdot}L^{-1}$ malt extract, $50g{\cdot}L^{-1}$ sucrose, $1.0 mg{\cdot}L^{-1}$ kinetin, and $8g{\cdot}L^{-1}$ agar). After four weeks of culture, 10, 21, 13, 5, and 7 somatic embryos and 2, 4, 2, 4, and 5 white callus cells (surrounding green somatic embryos) were obtained from Dongjeongkyool, Cheongkyool, Jikak, Miyagawa wase, and Haryejosaeng, respectively. After six weeks of culture, somatic embryos were obtained from cultured cells grown on MT basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), lactose ($70g{\cdot}L^{-1}$), and agar ($16g{\cdot}L^{-1}$). Over 60% of the somatic embryos from citrus cultivars native to Jeju developed into normal plants on MS basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), sucrose ($50g{\cdot}L^{-1}$), and agar ($8g{\cdot}L^{-1}$) after 10 weeks of culture. Normal plants were regenerated from two Citrus unshiu Marc. cultivars on MT basal medium supplemented with sorbitol (1.0 M), galactose (1.0 M), $GA_3$ ($1.0mg{\cdot}L^{-1}$), and Gelrite ($3g{\cdot}L^{-1}$). The absence of virus in plants generated from cultured ovules was confirmed by RT - PCR and antigen - antibody reactions. Therefore, virus - free Citrus cells can be obtained for breeding high - quality citrus cultivars using the biotechnological technique evaluated in this study.

• Journal of Korean Society of Forest Science
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• v.109 no.4
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• pp.413-420
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• 2020

We analyzed the growth characteristics of each cell line and acclimating substrate of Larix kaempferi plantlets regenerated from somatic embryos, with the goal of increasing the survival rate during the acclimation phase. Somatic embryos from three embryogenic cell lines (L14-66, L16-18, and L17-B4) were used, and the acclimating substrates were commercial soils for Larix species (Larix-Soil) and horticultural corps (Hort-Soil), Elle-pot, and Peat-plug. The average initial shoot and root length was shortest in L14-66 and longest in L17-B4. The average survival rate by cell line was highest (87.0%) in L17-B4 and lowest (64.3%) for L14-66. Survival rates by substrate were highest in Elle-pot (88.5%) and Peat-plug (88.9%). The initial shoot length of the L14-66 plantlets was highly correlated with survival rates in the Larix-Soil (r = 0.852), Hort-Soil (r = 0.692), and Elle-pot (r = 0.867) substrates, but not in Peat-plug with high total nitrogen content. The initial shoot length of the L17-B4 plantlets was not correlated with the survival rate in any of the substrates. The initial root length of the L14-66 plantlets was highly related to survival rates in the Larix-Soil (r = 0.986), Elle-pot (r = 0.846), and Peat-plug (r = 0.802) substrates, and the survival rate of the plantlets was higher as the initial root length was longer. The initial root length of the L17-B4 plantlets was related to survival rate only in the Larix-Soil (r = 0.896) and Elle-pot (r = 0.696) substrates. In conclusion, to increase the survival rate of plantlets, root length should be considered over shoot length, and it is recommended to use substrates with high nitrogen content, such as Peat-plug, or to add fertilizer, during the acclimating process. In addition, in order to increase the survival rate, lines with high initial growth should be developed.

• Journal of Plant Biotechnology
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• v.34 no.2
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• pp.139-144
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• 2007

Hypocotyl explants of Chinese cabbage (cvs. "Jeong Sang") produced transgenic calli on callus induction medium (MS salt, B5 vitamin, 5 mg/L acetosyringone, 1 mg/L 2,4-D, 3% sucrose, 400 mg/L cefotaxime, 100 mg/L paromomycin, pH 5.8) after cocultivation with strains of Agrobacterium tumefaciens (EHA101, LBA4404, GV3101) harboring the pPTN290 containing paromomycin-resistance gene as a selectable marker, and then they transferred to root induction medium (1/2MS salt, MS vitamins, 2% sucrose, 100 mg/L paromomycin, 100 mg/L cefotaxime, pH 5.8) and shoot induction medium (MS salt, B5 vitamin, 4 mg/L $AgNO_3$, 4 mg/L 6-benzyladenine, 3 mg/L alpha-naphthaleneacetic acid, 100 mg/L paromomycin, 100 mg/L cefotaxime, 3% sucrose, pH 5.8) in order. There was a significant difference in the frequency of transgenic calli depending on Agrobacterium strains. In particular, the highest frequency (6.1%) of transgenic calli was obtained from the hypocotyls cocultivated with EHA101 strains. Also, the frequency (%) of transgenic root and plants from each transgenic callus clone were obtained with 60.7% and 38.2% in EHA101, with 8.3% and 0% in LBA4404, with 20.5% and 85.7% in GV3101 strains, respectively. They were grown to maturity in a greenhouse and normally produced $T_2$ seeds. GUS histochemical assay for progeny ($T_2$) revealed that the transgenes was expressed in the plant genome, and progeny analysis from 7 independent transgenic events demonstrated that the transformants transmitted the transgene as a single or multiple functional locus.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.129-134
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• 2002

194 plantlets regenerated from leaf explants of boxthorn 'Cheonyang' were cultivated to investigate their morphological characteristics in the field for 2 years. Based on the morphology of leaves, 66.1% of them had elliptical type leaf, the same as that of mother plants, while 22.2% in oval type, 7.2% in obovate type, 2.6% in long-obvate type and 2.1% in lanceolate type. They were classified to 4 groups; group A was selected with both high fruit size and fruit yield, group B with only high fruit size, group C with larger or thicken leaf, and group D with multiple brenches. In comparision of production efficiency between the selected groups and mother plants, group A (99741, 99781, 99854, 99870 and 99886) were longer (2.1 to 2.7 mm) in length of fruits and higher in fruit production (15 to 30%) as compared to mother plants. Croup D (99797 and 99892) was higher in leaf production (7.2%) as compared to boxthorn 'CL1-48', which is the highest in leaf production among boxthorn veriaties.

• Journal of Plant Biotechnology
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• v.29 no.1
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• pp.45-49
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• 2002

The leaf and petiole segments of Farfugium japonica were cultured to investigate the influence of growth regulators on their callus induction and plant regeneration. The callus induction and growth showed a good response both leaf and petiole on MS media supplemented with 1∼2 mg/L 2,4-D and 1∼2 mg/L BA. Callus induction and growth were more effective in petiole segments than leaf one. The highest percentage of plant regeneration was obtained from 60-day-old calli on MS medium supplemented with 1 mg/L NAA and 2 mg/L BA. When subcultured to the same medium for about 60 days, multiple shoots were developed from regenerating callus. The shoots produced roots after transferring to rooting medium containing 0.5 mg/L IAA. The plantlets over 50 mm in height were successfully acclimatized in vermiculite, and the survival rate was over 95%.

• Journal of Korean Society of Forest Science
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• v.79 no.2
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• pp.109-114
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• 1990

The axillary buds of 15-year-old Tilia amurensis were cultured on Saito and Ide (IS), Murashige and Skoog (MS) media and woody plant medium (WPM) to establish an effective micropropagation method. Five levels of 6-benzylaminopurine (BAP) were tested. On IS medium and WPM addition of 1.0/l BAP enhanced shoot development and shoot elongation, whereas addition of 0.5/l BAP was effective on MS medium. A better results were obtained from WPM with 1.0/l BAP and MS with 0.1/l BAP. Developed shoots were subcultured on each basal media but with 0.2/l BAP, Multiple shoots were almost doubled in a month. Root formation could be enhanced at higher concentration of indole-3-butyric acid (IBA). Better rooting rate (83.3%) was achieved on a half-strength MS medium with 3.0 /l IBA. Regenerated plantlets were successfully transferred to soil. To investigate the clonal variation in shoot development and shoot elongation by axillary bud culturing, seven plus tree clones were tested, Clonal variation in tissue culturability among plus trees was recognized by the Duncan's multiple range test at the 5% level. Kang Won No. 12 showed the best response on WPM with 1.0/l BAP.

• Korean Journal of Plant Resources
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• v.25 no.2
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• pp.277-284
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• 2012

In the present study, the effects of plant growth regulators on adventitious shoot and root formation of various explants of $in$ $vitro$ seedlings of $Solanum$ $nigrum$ L. were investigated to determine the optimum conditions for the high-efficiency plant regeneration of this species. The formation of adventitious shoots was higher in leaf explants than in cotyledon, hypocotyl, or epicotyl explants at low concentrations (0.5~2.0 mg $L^{-1}$ ) of 6-benzylaminopurine (BAP). The number of adventitious shoots and the shoot length were also higher in both leaf and cotyledon explants. In particular, 2.0 mg $L^{-1}$ BAP was most effective for stimulating the induction and multiplication of adventitious shoots. In terms of root formation and root development from shoots that were separated from multiple shoots, indole butyric acid (IBA) and indole acetic acid (IAA) were more effective than ${\alpha}$-naphthalene acetic acid (NAA). The percentage of rooting as well as the number of roots per shoot (4.0), root length (7.82 cm), and shoot length (8.76 cm) was highest on MS media supplemented with 0.05 mg $L^{-1}$ IAA. Furthermore, 100% of the regenerated plantlets survived when transplanted to compost soil. These results suggest that leaf explants are the best source for the high-efficiency regeneration of $S.$ $nigrum$ L., and that 2.0 mg $L^{-1}$ BAP and 0.05 mg $L^{-1}$ IAA are the best conditions for shoot and root induction, respectively.

• Korean Journal of Plant Resources
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• v.23 no.1
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• pp.7-13
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• 2010

This experiment was conducted to establish the micropropagation of Calanthe discolor through multiple shoot formation from the culture of leaf, corm and root explants. Frequency of adventitious shoot formation from leaf explants was higher than those of corms and root explants. Frequency of adventitious shoot formation on medium with various concentrations of BA (0. 1.0, 3.0, and 5.0 mg/L) and NAA (0, 0.1, 0.5, and 1.0 mg/L) was tested. The maximun induction of adventitious shoot was obtained on half strength Murashige and Skoog (MS) medium supplemented with 3.0 mg/L BA and 1.0 mg/L NAA after 6 weeks of culture. Multiple shoots were transferred onto half strength MS medium with various concentrations of GA3 (0, 0.1, 0.5, 1.0, 3.0, and 5.0 mg/L). The number and length of multiple shoots on medium were highest on medium with 3.0 mg/L GA3. All the adventitious shoot grew well and rooted on half strength MS medium with 3.0 mg/L NAA. The plantlets were acclimatized up to 100% on sand with TKS-II or pearlite with TKS-II.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.79-79
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• 2019

홍띠(Imperata cylindrica 'Rubra') 식물자원의 기내대량증식과 재분화식물체의 순화체계를 구축하고자 기내 재분화에 적합한 식물재료부위, 생장조절물질을 조사하고, 재분화 유식물체로부터 적정 순화조건을 구명하였다. 기본배지로 MS (Murashige and Skoog, 1962) 배지를 사용하였고, 배양은 $26{\times}2^{\circ}C$, $25{\mu}mol/m^2/s$, 14h/10h (day/night) 광조건 하의 배양실에서 수행하였다. 캘러스 형성은 뿌리 끝, 줄기절편, 생장점 부위 중생장점 부위에서 가장 양호하였고, 이 생장점 조직에 0.1 mg/L의 2,4-D와 2 mg/L의 BA를 처리하였을 때 양호하였다. 캘러스 증식은 0.1 mg/L의 2,4-D와 0.05 mg/L의 BA 배지, 0.05 mg/L의 2,4-D와 0.5 mg/L의 BA를 첨가한 배지 중 0.1 mg/L의 2,4-D와 0.05 mg/L의 BA 배지에서 양호하였고, 이들 캘러스로부터 신초 재분화는 0.01 mg/L의 NAA와 2 mg/L의 BA 처리에서 양호하였다. 초기 치상으로부터 실제 경과시간은 캘러스 유도에 19주간(2018. 03. 18~07. 27), 캘러스 증식 9주간(2018. 07. 27~09. 28), 신초 유도 11주간(2018. 09.28~12. 14), 순화에 10주간(2018. 12. 14~2019. 02. 23)에 걸쳐 진행하였으나 확립된 배양계를 적용하면 캘러스 유도 4주, 캘러스 증식 3주, 신초유도 및 증식 ４주, 순화 ７주 정도가 소요될 것으로 계측되었다. 순화는 다경줄기 형성후 MS배지를 멸균한 상토(버미큘라이트) 또는 종이포트로 교체하여 재분화식물체를 배양병에서 7주간 배양하고, 7주후에 배양병 뚜껑을 1/10 정도 1차 개방하여 1주일 후 3/10 정도 개방하여 2주간 경과한 후 컵포트(직경 6 cm)에 이식하여 성공적으로 활착시켰다.