• Title/Summary/Keyword: 기내배양

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Transformation of Populus alba $\times$Populus glandulosa Using Phosphinothricin Acetyltransferase Gene (Phosphinothricin acetyltransferase 유전자를 이용한 현사시의 형질전환)

  • 오경은;양덕춘;문흥규;박재인
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.163-169
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    • 1999
  • This study was conducted to produce herbicide resistant plants by transferring phosphinothricin acetyltransferase (PAT) gene into Populus alba $\times$ Populus glandulosa No .3 using Agrobacterium tumefaciens MP 90/PAT. Leaf segments from in vitro grown shoots of hybrid poplar No. 3 were soaked in a AB medium containing Agrobacterium tumefaciens MP 90/PAT for 10 min and cocultivated for 2 days on MS medium containing 1.0 mg/L 2,4-D and 0.2mg/L kinetin (CIM). Putative transformed calli could be selected after cocultivation of leaf segments on CIM supplemented with 50mg/L kanamycin and 500mg/L cefotaxime for 3 weeks. The selected calli were cultured on CIM supplemented with 50 mg/L kanamycin and 500 mg/L cefotaxime for 5~8 weeks before transfer to WPM containing 1.0mg/L zeatin, 0.1mg/L BAP, 50 mg/L kanamycin and 500mg/L cefotaxime for shoot regeneration. Shoots were regenerated from the callus after 4 week cultivation, and the regenerants were grown on the same medium for 7~l0 weeks. The plants rooted on 1/2 WPM containing 0.2 mg/L IBA and 50 mg/L kanamycin. To confirm the gene insertion into plants, GUS activity was detected by histochemical assay in the transformed plants. Finally, the presence of both NPT II and PAT genes from the transgenic plants were confirmed by PCR amplification with the gene specific primers and subsequent PCR-Southern blot with DIG-labeled PAT gene probe. After acclimatization in pots for 4 weeks, the plants were sprayed by 3 mL/L of Basta to test resistance to the herbicide. The transgenic plants remained green, whereas all the control plants died after one week.

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Effects of Light Quality and Lighting Type Using an LED Chamber System on Chrysanthemum Growth and Development Cultured In Vitro (LED Chamber System을 이용한 광질 및 광조사 방법 제어가 국화 배양소식물체의 생장에 미치는 영향)

  • Heo, Jeong-Wook;Lee, Yong-Beom;Chang, Yu-Seob;Lee, Jeong-Taek;Lee, Deog-Bae
    • Korean Journal of Environmental Agriculture
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    • v.29 no.4
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    • pp.374-380
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    • 2010
  • This experiment was carried out to investigate the effect of light qualities and lighting types provided by LED Chamber System which designed by Rural Development Administration on growth and development of Chrysanthemum (Dendranthema grandiflorum L., cv. 'Cheonsu') plantlet cultured in vitro. The explants of single-node cuttings were exposed to monochromic or mixture radiation of blue, red, or green under continuous and intermittent lighting for 42 days. The intermittent lighting of 20 sec. on and off per minute significantly stimulated shoot elongation with lower number of internodes compared with continuous lighting treatments. However, continuous blue, red, or green light gave greater dry weight comparing the intermittent lighting, and the lowest weight was recorded at the continuous fluorescent lamp. Otherwise, the plantlet growth in dry weight or leaf area was inhibited by the green light controlled at 50 times intermittence but internode elongation was significantly increased. These results showed that the plantlets were successfully grown under the LED Chamber System controlled with different light qualities and lighting types. Quantitative growth of the plantlets was improved under the shorter photoperiod with a intermittent lighting cycle compared with continuous lighting using fluorescent lamps. It is concluded that the growth and development of in vitro plantlets such as single-node cuttings can be achieved by the controlling of light quality or lighting type during the photoperiod per day with a lower electric cost compared with conventional continuous lighting system.

Micropropagation of Calanthe discolor Lindl. through Induction of Multiple Shoots from Axillary Bud Culture (새우난초(Calanthe discolor Lindl.)의 액아배양으로부터 다신초 형성을 통한 대량증식)

  • Lim, Ju Hong;Chung, Mi Young;Kim, Chang Kil;Lim, Ki Byung;Chung, Jae Dong
    • FLOWER RESEARCH JOURNAL
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    • v.16 no.4
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    • pp.239-246
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    • 2008
  • This study was conducted to establishment of in vitro micropropagation through induction of multiple shoots from axillary bud culture in Calanthe discolor Lindl. Shoots initiation from axillary bud was the most effective on half strength MS medium supplemented with $0.5mg{\cdot}L^{-1}$ NAA and $2.0mg{\cdot}L^{-1}$ TDZ during four weeks of dark culture followed by culture under a 16-h photoperiod. Multiple shoots (12.5 shoots per explant) were proliferated on half strength MS medium containing $4.0mg{\cdot}L^{-1}$ TDZ and $2.0mg{\cdot}L^{-1}$ IBA. On the other hands, the abnormally emerged shoots during the multiple shoot proliferation stage were recovered to normal shoots on half strength MS hormone free medium. Multiple shoots were well elongated and rooted on half strength MS medium with $0.1mg{\cdot}L^{-1}$ NAA. The plantlets were acclimatized up to 100% on TKS substrate after pretreating with $10mg{\cdot}L^{-1}$ NAA for 30 min. and these plantlets showed good growth as well.

Optimal Culture Conditions for Masspropagation of Gametophytes and Sporophytes of Pyrrosia linearifolia by Tissue Culture (조직배양을 이용한 우단일엽의 대량번식을 위한 전엽체와 포자체의 적정 배양조건)

  • Shin, So Lim;Lee, Cheol Hee
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.3
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    • pp.179-185
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    • 2010
  • This study was carried out to investigate the optimal culture conditions for gametophytes growth and sporophytes regeneration of Pyrrosia linearifolia in order to provide for the masspropagation system foundation of Pyrrosia linearifolia using their life cycle. Among many different media, 2MS medium was most effective in prothallus proliferation. Prothallus growth was promoted as the total concentration of nitrogen sources increased, and the best result was observed on 120 mM nitrogen. The best concentration of sucrose was 3%. The addition of 5~20 mM IAA, NAA, BA and kinetin promoted the propagation of prothallus. But 2iP demonstrated the most inhibitory effect on prothallus proliferation. Gametophytes shaking-cultured with liquid medium showed similar growth with solid medium and normal formation of reproductive organs. Shoot regeneration was most effective on 1/8MS medium, but growth was promoted on 1/2MS medium. For promotion of shoot regeneration and growth, the suitable concentrations of sucrose and $NaH_2PO_4$ were 1% and $50{\sim}100mg{\cdot}mL^{-1}$ in 1/8MS medium, respectively.

Plant Regeneration and Genetic Diversity of Regenerants from Seed-derived Callus of Reed (Phragmites communis Trinius) (갈대(Phragmites communis Trinius) 성숙종자를 이용한 기내 식물체 재분화와 재분화체의 유전적 다양성)

  • Ryu, Jaihyunk;Kim, En-Hwan;So, Hyun-Su;Chung, Mi-Young;Song, Won-Seob;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.26 no.2
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    • pp.320-327
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    • 2013
  • This study was carried out to address an efficient in vitro regeneration system from seed-derived callus of Phragmites communis, and to evaluate genetic variations of the regenerants using ISSR markers. Shoot regeneration via calli was greatly influenced by N6 medium compared with MS medium, and plant regeneration frequency was 90% in N6 supplemented with BA 0.25 mg/L and BA 0.5 mg/L. According to ISSR analysis of the thirty regenerants, out of 94 loci detected overall, 16 were identified to be polymorphic with a rate (PR) of 17.0%. The mean gene diversity (h) of different in vitro condition was 0.03 and ranged from 0.008 for N6 with BA 5 mg/L, to 0.040 for MS with IAA 0.1 mg/L+kinetin 2 mg/L. The results indicate that the regenerants have a low genetic variation, and ISSR analysis is effective to detect genetic variation of regenerants.

Effect of LEDs on shoot multiplication and rooting of rare plant Abeliophyllum distichum Nakai (희귀 수종 미선나무(Abeliophyllum distichum Nakai.)의 기내 증식 및 발근에 미치는 LED (light emitting diode) 효과)

  • Lee, Na Nyum;Choi, Yong Eui;Moon, Heung Kyu
    • Journal of Plant Biotechnology
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    • v.41 no.2
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    • pp.94-99
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    • 2014
  • This study was conducted to elucidate the effect of light sources and explant types on in vitro shoot multiplication and rooting of a rare and endangered plant Abeliophyllum distichum. Both apical buds and axillary buds were used as explants under 4 different light sources, cool white florescent light (F), 100% blue light-emitting diode (LED) (B), 50% blue and 50% red LED mixture (BR), and 100% red LED (R). Clear difference was observed in terms of shoot proliferation by light sources types but not by position-dependent explant types. Multiple shoot induction rates were enhanced under both B and BR light sources. Spontaneous rooting was induced in shoot induction medium under B light source. Both the rates of rooting and numbers of roots per explant were higher in apical bud explants compared to axillary bud explants. Interestingly R light source stimulated shoot elongation but inhibited root development. Therefore, our results suggest that the use of apical bud explants under B or BR light sources is suitable for in vitro micropropagation of a rare and endangered plant species, Abeliophyllum distichum.

Selection of Heat-tolerant Potato Clones by In Vitro Tuberization from True Potato Seeds (감자 眞正種子의 器內小塊莖 形成에 의한 耐暑性 系統 選拔)

  • 김현준;김영화;유승렬;김병현;김정간
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.77-81
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    • 1997
  • For the early selection of heat-tolerant clones, the true potato seeds of 750 clones were obtained by several cross combinations : $CIP\;575015\;{\times}\;katahdin,\;CIP\;575015\;{\times}\;B6603-6,\;84\;I\;35-4\;{\times}\;katahdin,\;CIP\;575015\;{\times}\;NookSack,\;and\;CIP575015\;{\times}\;Superior.$ The ratios of in vitro tuberization at 3$0^{\circ}C$ were decreased by 43% in all cross combinations compared with at 2$0^{\circ}C$. In particular, tuberization rate of $CIP575015\;{\times}\;katahdin$ cross at 3$0^{\circ}C$ was only 21%. On the other hand, the rate of tuberization of $CIP575015\;{\times}B6603-6$ cross was 58 % at 3$0^{\circ}C$, so this cross combination was thought to be good heat-tolerant clone. To screen the heat-tolerant clones in lower land during high temperature period, microtubers were cultivated in Kangnung, and the characters of tubers and productivity were examined. Among screened clones, one clone at 2$0^{\circ}C$ and three clones at 3$0^{\circ}C$ were shown to be heat-tolerant. The yield of 89ML75-8 was 88% more than that of Superior, and dry weight rate of 89 ML64-11 was 18.8%. Therefore, 89ML64-11 clone was considered as a good cultivar for processing.

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Development of Aspceptic Seedling by In Vitro Germination in Lacquer Tree Seed (옻나무 종자의 기내 발아에 의한 무균묘 육성)

  • 두홍수;이호림;권태호;양문식
    • Korean Journal of Plant Resources
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    • v.13 no.1
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    • pp.48-53
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    • 2000
  • Lacquer tree can be proliferated by root or stem cutting, and seed. In case of proliferation by seed, however, the germination rate is very low. Thus, the present study was carried out to obtain aspceptic lacquer plant in vitro from seed because natural tissue culture was highly defiled by unknown fungi and bacteria. First seed grading on distilled water was 50.7% and second seed grading was 20.8% after 98% sulfuric acid treatment for 2 hours. Removal of inner seed coat was higher with 32.4% than non-removal of outer seed coat and removal outer seed coat in rooting rate. In germination rate according to pre-treatment, growth regulators were not effective at all, but sulfuric acid was effective a little with 3%. Removal outer seed coat was increased about 4%, that germinated about 10% in MS medium supplemented with 1.0mg/L BA and 0.05mg/L NAA, 1.0mg/L BA. Lacquer tree seeds germinated after 10 days in MS medium, and aspceptic seedling of lacquer tree were obtained after 3 weeks in vitro. Germination rate, however, was lower about 10%.

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Effect of explant's position and culture method on shoot proliferation and micro-cuttings for a rare and endangered species, Abeliophyllum distichum Nakai (희귀 및 멸종위기 식물 미선나무(Abeliophyllum distichum Nakai)의 절편위치 및 치상방법에 따른 기내증식 및 미세삽목)

  • Lee, Na Nyum;Kim, Ji-Ah;Kim, Yong-Wook;Choi, Yong Eui;Moon, Heung Kyu
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.228-234
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    • 2015
  • Using either the apical or axillary bud of the endangered species Abeliophyllum distichum Nakai, we tested the effect of bud position and culture method on shoot proliferation and rooting. In shoot proliferation, the axillary bud explant was more effective than the apical bud and the effect was fostered by BA treatment, whereas no differences were observed in shoot elongation by the explant position. Spontaneous rooting was observed in the MS basal medium and resulted in conspicuous differences in the explant position : more than 80% in apical bud explant and 28% in axillary bud explant was achieved, respectively. The positional effects were also observed in BA pre-treatments: generally vertical culture method appeared to be better in shoot proliferation, growth, and rooting than that of the horizontal culture method regardless of the BA pre-treatment duration. The highest shoot multiplication was achieved through the vertical culture method with axillary bud explant, whereas the best shoot elongation and rooting was obtained using the vertical culture method with the apical bud explant. Apical bud explant was superior to axillary bud explant in ex vitro micro-cuttings and revealed a significant difference in shoot growth and root development. The above results suggest that explant position and culture method influence the efficiency of micropropagation for a rare and endangered plant Abeliophyllum distichum.

Plant Regeneration Via Direct Adventitious Roots from Free Root Segments of Ulmus davidiana Planch (당느릅나무(Ulmus davidiana Planch) 부정근 절편으로 부터 부정아 유도를 통한 식물체 생산)

  • Kim, Ji-Ah;You, Xiang-Ling;Ahn, Chang-Ho;Lee, Jae-Seon;Choi, Yong-Eui
    • Journal of Korean Society of Forest Science
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    • v.96 no.1
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    • pp.83-88
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    • 2007
  • Micropropagation of Ulmus davidiana Planch was established via adventitious shoot formation from the segments of adventitious roots. Adventitious roots were produced directly from root segments of seedlings on a 1/2 SH medium plus various concentrations of IBA. The maximum growth of adventitious roots was observed in the presence of 2.0 mg/L IBA. After the segments of adventitious roots were cultured on various cytokinins (zeatin, 2-iP, BA, kinetin) and cytokinins plus auxin (IBA), formation of adventitious shoot was investigated. Among cytokinin treated, kinetin was the most effective on both adventitious shoot induction and number of shoots. Especially, 2.0 mg/L kinetin was the best to increase adventitious shoot induction (95.8%) and a number of shoots (8.4). Adventitious shoots were rooted on 1/2 WPM medium and the plantlets were acclimated 100% on composed soil (peatmoss : vermiculite 1 : 1).