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Effect of Explant Types, Auxin Concentration and Light Condition on In Vitro Root Production and Alkaloid Content of Rauvolfia serpentina (L.) Benth. ex Kurz

  • Yahya, Andi Fadly;Hyun, Jung-Oh;Lee, Jae-Ho;Jung, Myung-Suk
    • Journal of Korean Society of Forest Science
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    • v.96 no.2
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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.

High-frequency regeneration of plants in vitro from seedling-derived apical bud explants of Tilia mandshurica Rupr. & Maxim

  • Kim, Tae-Dong;Kim, Nam-Ho;Park, Eung-Jun;Lee, Na-Nyum
    • Journal of Plant Biotechnology
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    • v.48 no.1
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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.

Callus Induction and Plant Regeneration in Angelica koreana MAX. (강활(Angelica koreana MAX.) 조직배양을 통한 캘러스 유도와 식물체 재생)

  • 장기원;민경수
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.39 no.6
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    • pp.537-541
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    • 1994
  • This study was conducted to investigate the possibility of callus induction and plant regeneration from immature inflorescence, stem and petiole of A. koreana MAX. which is worth enough to be used as food and medicine. The callus induction and its proliferation was best when immature inflorescence segments were placed on MS medium supplemented with 2, 4-D 2mg / l. The white and compact embryogenic callus on the surface of dark yellow and soft callus which was induced from immature inflorescence segments came into being only on MS medium with 2, 4-D 1mg /l and 2mg /l, but didn't come into being on the other ones. The shoot came into being effectively from callus derived from immature inflorescence on MS medium mixed 2, 4-D 0. 1mg /l with Kinetin 1mg /l, and 2, 4-D 0.5mg /1 with Kinetin 2mg /l. Immature inflorescence was most appropriate material for callus induction and plant regeneration.

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Plant Regeneration by Anther Culture of Lilium asiatic hybrid 'Gran Paradiso' (아시아틱 백합 ( Lilium asiatic hybrid 'Gran Paradiso' )의 약배양에 의한 식물체 재생)

  • 고정애
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.1-6
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    • 1999
  • In order to obtain plantlet derived by anthers, the anthers of Lilium asiatic hybrid 'Gran Paradiso' were cultured on Murashige and Skoog's medium supplemented with various combinations of auxin and cytokinin. The most suitable pollen stage of anther culture for the callus induction was 3 days before anthesis at the early to late binucleate stage. Organogenic calli were induced on MS medium supplemented with 5.0 mg/L 2,4-D alone and the combination of 1.0 mg/L 2,4-D and 1.0 mg/L kinetin, however, the combination of NAA and BA was more effective than that of 2,4-D and kinetin on plant regeneration through organogenesis. Shoots were formed from the induced callus on the medium with 0.5 mg/L NAA and 1.0 mg/L BA after 180 days of culture. Multiple shoots with 3-4 leaves, roots, and bulblets were formed on the medium with the combination of 2.0 mg/L NAA and 2.0 mg/L BA after 250 days of culture. The chromosome from root tip of the regenerated plantlet showed the diploid (2n=2x=24). Diploid plants were transferred to the pots and all plants were flowered in two years.

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Effects of Carbon, Nitrogen Sources and pH on Direct Somatic Embryogenesis in Liquid Culture of Rehmannia glutinosa Lib. (지황의 액체배양에서 탄소원.질소원 및 pH가 직접 체세포배 형성에 미치는 영향)

  • Chae, Young-Am;Park, Ju-Hyun
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.1
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    • pp.1-6
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    • 1999
  • Basic informations for direct somatic embryo formation in Rehmannja glutinosa Lib. were obtained in 500ml erlenmyer flask. The ratio of ammonium to nitrate nitrogen of 825(mg/l) : 1900(mg/l) was proper condition for somatic embryo formation from stem and petiole explants and 3% sucrose was the most effective carbon source. Full strength MS medium with 2mg/l BA was better than LS medium for somatic embryogenesis. The initial pH 5.7 of medium(full strength MS with 2.0mg/l BA and 0.1mg/l NAA) was good for embryo production. Potassium ion was taken up rapidly within 2 weeks. while $Ca^{++}$ and $Mg^{++}$ ion contents were almost constant during culture period. Sucrose hydrolysis occurred throughout the culture, while glucose and fructose were absorbed simultaneously from the third week of culture.

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Plant Regeneration via Adventitious Shoot Formation in Platycodon grandiflorum (Jacq. A. DC.) (도라지 (Platycodon grandiflorum (Jacq.) A. DC.) 부정아 형성을 통한 식물체 재분화)

  • Kim, Ju Young;Na, Hyun Sun;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.330-334
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    • 2017
  • To investigate optimal conditions for plant regeneration in Platycodon grandiflorum (Jacq. A. DC.).Both leaf and hypocotyl explants were cultured on Murashige& Skoog's (MS) medium supplemented with combinations of 0.1, 0.5, 1.0, or 2.0 mg/L cytokinins (BA and kinetin) and 1.0 mg/L 2,4-D for 6 weeks, respectively. According to the type of explant, the total shoot organogenesis (56.38%) in leaf explants was higher than in hypocotyls (28.20%). In comparison with kinetin and BA for the plant regeneration, the frequency (70.38%) of leaf explants was higher in combination with kinetin and 2,4-D than of BA with 2,4-D (42.38%), whereas the frequency (35.56%) of hypocotyls explants was higher in BA combination than kinetin combination (20.83%). Thehighest frequency (94.20%) was observed from the cultures of leaf explants on the MS medium supplemented with 1.0 mg/L kinetin and 1.0 mg/L 2,4-D. Upon transfer onto 1/2 MS basal medium containing 3% sucrose, shoots developed into plantlets with roots, and were well grown in soil in the greenhouse. These results lead us to speculate that the optimization of culture conditions was responsible for the mass propagation from in vitro cultures of Platycodon grandiflorum (Jacq. A. DC.).

Efficient Plant Regeneration from Mesophyll Protoplast of Arabidopsis thaliana and Morphological Characterization of Regenerants (애기장대 (Arabidopsis thaliana)의 엽육원형질체로부터 효율적인 식물체 재분화와 이들의 형태적 특성)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.127-132
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    • 1999
  • Protoplasts were isolated from the leaf mesophyll tissue of in vitro 4-weeks-old Arabidopsis thaliana and cultured in MS liquid medium supplemented with 2.0 mg/L NAA, 0.5 mg/L BAP and 9% mannitol in the dark at $25^{\circ}C$. When protoplast-derived microcolonies were dehydrated, the frequency of callus induction enhanced approximately 7-fold higher compared with non-dehydrated microcolonies in CP medium. Fifty callus lines were selected from dehydrated microcolonies. Shoots were efficiently initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium supplemented with 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for 4 weeks. Shoot regeneration frequencies (calli regenerating at least one shoot) were 3.5%~56%. Histological observations of shoot forming callus revealed that tracheary elements initiated from inner compact cells, and that meristemoids developed to shoot primordia and shoots. Roots were induced from these regenerating shoots on MS medium without phytohormones. These regenerants were successfully transplanted into potting soil. Morphological characterization of 50 protoplast-derived plants showed that the frequency of normal type was 78%.

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Isolation and Structural Identification of Antioxidant Substances from Ethyl Acetate Extract of Conyza canadensis (망초(Conyza canadensis) Ethyl Acetate 추출물의 항산화성 물질의 분리와 동정)

  • Hyun Sook Song
    • Journal of Naturopathy
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    • v.12 no.1
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    • pp.7-15
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    • 2023
  • Background: As a result of analyzing the components of wild Conyza canadensis, it contains physiologically active ingredients, so it is necessary to identify the compound. Purposes: It was to study the compound's molecular structure; a previous study showed that C. canadensis contains antioxidant substances. Methods: The ultrasonic pulverized lysate of C. canadensis stem and leaves was first extracted with 90% methanol and then five organic solvents. Next, the extracts was fractionated by HPLC, LC/MS chromatography, and NMR analyzers identified the molecular structure. Results: 100 g of dry C. canadensis was sonicated in 90% methanol and concentrated under reduced pressure to 11.96 g of a crude extract. Then, this crude was extracted with five types of solvents to obtain 123.8 mg of n-hexane, 448.2 mg of dichloromethane, 1047.7 mg of ethyl acetate (EA), 2563.8 mg of butanol, and 7.04 g of water. The EA extracts were fractionated by LC-MS and then re-fractionated to obtain F1 to F20. Next, the F15 was further fractionated to obtain nine fine fractions. Finally, the F17 fraction was re-fractionated to obtain ten fine fractions. As a result of LC-MS and NMR spectrometer analysis of the F15-7, the structure of this compound was confirmed as 3,5-dicaffeoylquinic acid. As a result of examining the structures of the F17-4 and F17-5 fractions, Quercetin-3-o-β-galactose was identified. In addition, the form of the F17-10 was confirmed to be 1,3,4-tri-caffeoylquinic acid. Conclusions: This study demonstrated that C. canadensis contained phenolic antioxidants, and its utilization may be expected.

Comparison of Liquid Chromatography-Mass/Mass Spectrometry (MS) and Gas Chromatography-MS for Quantitative Analysis of Indole-3-acetic acid and Indole-3-butyric acid from the Concentrated Liquid Fertilizer (Liquid Chromatography-Mass/Mass Spectrometry (MS)와 Gas Chromatography-MS를 이용한 농축 액상 비료제품 중 Indole-3-acetic acid 및 Indole-3-butyric acid 정량분석능 비교)

  • Kim, Jin Hyo;Park, Jong Min;Choi, Geun-Hyoung;Park, Yun-Ki;Im, Geon-Jae;Kim, Doo-Ho;Kwon, Oh-Kyung
    • Journal of Applied Biological Chemistry
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    • v.56 no.1
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    • pp.53-57
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    • 2013
  • In here, we investigated the quantitative analysis method of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) with liquid chromatography-mass/mass spectrometry (LC-MS/MS) or gas chromatography-MS. Two ways of clean-up process were investigated for LC-MS/MS instrumental analysis of IAA, but both a simple dilution and hydrophile-lipophile balance (HLB) solid phase extraction (SPE) were not met the optimal recovery rates for quantitative analysis. On the other hand, the clean-up method for GC-MS was finally optimized through HLB-SPE from 250-folds diluted sample and methylation with trimethylsilyl chloride in methanol for 4 h. The limit of detection for methyl ester of IAA and IBA were both 1.4 mg/L, and recovery rates showed 93-107% from the concentrated liquid fertilizer.

Effects of Different Natural Extracts and Plant Growth Regulators on Plant Regeneration and Callus Induction from Pseudobulbs Explants through in vitro Seed Germination of Endangered Orchid Bulbophyllum auricomum Lindl. (멸종 위기에 처한 Bulbophyllum auricomum Lindl. orchid의 시험관 내 종자 발아를 통한 구근 절편체의 식물 재생 및 캘러스 유도에 대한 천연 추출물 및 식물 성장 조절제(PGR)의 효과)

  • Aung, Win Theingi;Bang, Keuk Soo;Yoon, Seo A;Ko, Baul;Bae, Jong Hyang
    • Journal of Bio-Environment Control
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    • v.31 no.2
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    • pp.133-141
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    • 2022
  • Bulbophyllum auricomum Lindl. is a rare orchid and has flowers with an attractive fragrance. The present study investigated the tissue culture method for micropropagation. Capsules derived from artificial self-pollination were obtained for the best seed germination in MS basal medium. Plant growth regulators (1.0 mg·L-1 of BAP and 2.0 mg·L-1 of NAA) were affected by callus induction from subcultured pseudobulb explants. For the callus subculture, different natural plant extracts were tested in 11 treatment media. Among them, MS medium with 150 mL·L-1 of coconut water was generally effective in fresh weight (1.75 ± 0.08) and (3.01 ± 0.20) of callus proliferation and PLBs induction at 1 and 2 months, respectively, followed by an MS combination of 30 g·L-1 of banana and 20 g·L-1 of potato extract. The results of a comparative study of different MS mediums containing plant growth regulators with a natural extract combination and MS medium supplemented with natural extract only showed that MS medium supplemented with a combination of natural extracts (150 mL·L-1 of coconut water) and plant growth regulators (2.0 mg·L-1 of BAP and 1.0 mg·L-1 of NAA) obtained the highest shoot regeneration (3.37 ± 0.17) and (6.41 ± 0.68) after 1 month and 2 months of culturing, respectively.