• Title/Summary/Keyword: ex vitro

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설페이티드 다당류의 실험실적 효능 검색과 동물내에서의 비교

  • 김영식
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.76-76
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    • 1993
  • 헤파린은 혈액응고계에서 antithrombin III 존재하에서 thrombin과 factor Xa의 작용을 억제함으로써 항응고제로서 작용을 한다. 심정맥혈전증 등 임상에 응용성이 높지만 장기적 사용시 혈소판 감소효과, 출혈, 골다공증 등의 부작용이 나타나고 있다. 본연구는 식물성 생약으로부터 당을 분리 정제하여 화학적으로 sulfation시켜 in vitro와 ex vivo에서 항응고활성을 비교하였다. 우선적으로 aPTT를 측정하여 응고시간의 연장을 시키는 다섯종류의 식물생약을 선택하였고 이 중에서 청호(Artemisiae apiaceae)로부터 산성당을 분리하여 pyridine과 chlorosulfonic산으로 sulfation 시켰율 때 in vitro상에서 항응고활성은 sulfation전에 비해 두드러지게 증가하였다. 농도를 달리 하여 실험동물에 투여시 응고시간의 연장 역시 비슷한 양상을 보여주었다. Thrombin 억제는 발견되지 않았지만 sulfate기와 항응고 활성과는 관계가 있는 것처럼 보였다.

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Silver nitrate and silver-thiosulphate mitigates callus and leaf abscission during Shisham clonal micro-propagation

  • Raturi, Manoj Kumar;Thakur, Ajay
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.173-178
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    • 2021
  • Basal callus formation and leaf abscission is a problem in clonal micropropagation. We have described an in vitro clonal propagation protocol of Dalbergia sissoo Roxb (shisham) 'FRI-14' in which AgNO3 played important role not only in mitigating problem of leaf abscission and basal callus, but also improved shoot induction and multiplication. Best induction and shoot multiplication was obtained on MS media with 1.5 mg/l 6-BAP and 10 mg/l AgNO3 and half-strength MS media with 0.5 mg/l 6-BAP, 2 mg/l AgNO3 and 50 mg/l Adenine sulphate whereas best ex vitro rooting was obtained with 200 mg/l IBA in pulse treatment.

Effects of in vitro culture types on regeneration and acclimatization of yellow poplar (Liriodendron tulipifera L.) from somatic embryos

  • An, Chan Hoon;Kim, Yong Wook;Moon, Heung Kyu;Yi, Jae Seon
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.110-118
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    • 2016
  • We compared germination efficiency for somatic embryos (SE) of Liriodendron tulipifera using semi-solid (SS), temporary immersion bioreactors (TIB), and continuous immersion bioreactors (CIB) to produce vigorous plants. The bioreactors were designed to be immersed in liquid media with plantlets with an adjustable immersion time. TIB and CIB improved germination rates up to 80.86% and 95.21%, respectively, however, CIB produced more hyperhydric plantlets than TIB. The height of plantlets in TIB was significantly higher than for those in CIB. Fresh weights of plantlets grown in CIB of were significantly lower than for those grown in TIB. The lowest chlorophyll concentration was found in in vitro plantlets from CIB. We examined abnormally developed leaves, stems, and apical zones of in vitro plantlets that were produced in CIB. Among the three types, SS showed the highest stomatal density and the shortest stomatal length in in vitro plantlets. After acclimatization, plants from CIB exhibited the lowest values in biomass, such as height, root collar diameter, leaf fresh weight, leaf length, leaf width, petiole length, petiole diameter, and leaf area. Photosynthesis and transpiration rates of ex vitro plants were not significantly different among the three culture types, but stomatal conductance was higher in TIB than in the SS and CIB. Therefore, the results suggest that TIB is the preferable bioreactor to improve in vitro plantlet regeneration of L. tulipifera. TIB-originated plants showed higher growth rate than SS and CIB after transferring to soil.

Antibacterial potential of the extracts derived from leaves and in vitro raised calli of medicinal plants Pterocarpus marsupium Roxb., Clitoria ternatea L., and Sanseveiria cylindrica Bojer ex Hook

  • Shahid, M;Shahzad, A;Anis, M
    • Advances in Traditional Medicine
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    • v.9 no.2
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    • pp.174-181
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    • 2009
  • Pterocarpus marsupium, Clitoria ternatea, and Sanseveiria cylindrica are some of the important and endangered medicinal plant species of India. Despite of medicinal properties, antibacterial potential of the plants have not yet been explored. The present study was designed to optimize the in vitro technique for micropropagation and to screen the extracts from leaves and in vitro raised calli for antibacterial properties. Excised leaf-explants from the parent plants were surface sterilized and cultivated on Murashige & Skoog's (MS) medium containing $N^6$-benzyladenine (BA) in concentrations of 1, 2, 5, and $10{\mu}M$. Optimal growth of calli was noticed at a concentration of $5{\mu}M$, therefore the extracts from calli grown at this concentration were further studied for antibacterial activity. Both alcoholic and aqueous extracts from leaves of respective plants, and their in vitro raised calli were tested for antibacterial activity by agar well diffusion method against a range of Gram-positive and Gram-negative bacteria. Aqueous extracts showed antibacterial activity against limited number of bacterial species; notably the extracts of C. ternatea which showed antibacterial activity against Streptococcus pyogenes, Bacillus subtilis and Bacillus cereus. Alcoholic extracts of all three plants showed antibacterial activity against a wider range of bacteria. Among the Gram-positive bacteria, extracts from C. ternatea showed strong antibacterial activity against Bacillus spp., whereas the extracts of S. cylindrica showed good antibacterial potential for Staphylococcus aureus, S. epidermidis and S. pyogenes. The extracts from all three plants showed antibacterial activity against Gram-negative bacteria, including, Salmonella spp. and Shigella dysenteriae; organisms causing enteric fever and dysentery. In most of the cases, the extracts from respective calli showed comparable, and in some cases better, result in comparison to the extracts from parent leaves. To the best of our knowledge this is the first preliminary report on antibacterial potential, especially through calli extracts, of these plants; and in vitro cultivation of the explants may be used to obtain phytotherapeutic compounds.

Plant Regeneration from Leaf Explants of Kalanchoe daigremontiana Hamet & Perrier

  • ;Kim, Teh-Ryung;In, Jun-Gyo;Yang, Deok-Chun;Choi, Kwan-Sam
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.5
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    • pp.293-298
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    • 2006
  • Optimum culture conditions for high frequency plant regeneration from leaf explants of Kalanchoe daigremontiana Hamet &Perrier were established. Shoot regeneration was achieved from leaf explant cultures using MS medium supplemented with indole-3-acetic acid (IAA) and thidiazuron (TDZ) or benzyladenine (BA). Percent regeneration was influenced by plant growth regulators and source of explants. MS medium supplemented with TDZ (1.0 mg/l) and IAA (0.4 mg/l) was the most effective, providing shoot regeneration for 76.7 % of ex vitro leaf explants associated with a high number of shoots per explant (9.5 mean shoots per explant), whereas 100% shoot regeneration associated with 12.4 shoots per explant occurred from in vitro leaf explants on the same medium. Clusters of shoots were multiplied and elongated on MS medium containing several concentrations of BA. MS medium supplemented with 0.25 mg/l BA was proved as the most effective shoot elongation medium. Elongated shoots (2-3 cm) were rooted at 100% on half-strength MS medium. Rooted plantlets were then transferred to potting soil. Regenerated plants were established in the soil with 90% success.

In vitro propagation of Bambusa nutans Wall. ex Munro through axillary shoot proliferation

  • Negi, Divya;Saxena, Sanjay
    • Plant Biotechnology Reports
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    • v.5 no.1
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    • pp.35-43
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    • 2011
  • This communication describes for the first time an efficient and reproducible protocol for large-scale multiplication of Bambusa nutans. Nodal segments collected from field-grown clumps and cultured on Murashige and Skoog (MS) medium supplemented with $4.4{\mu}M$ benzylaminopurine (BA) and $2.32{\mu}M$ kinetin (Kin) gelled with 0.2% gelrite yielded 80% aseptic cultures with 100% bud-break. The in vitro-formed shoots obtained after bud-break were successfully multiplied in MS liquid medium supplemented with $13.2{\mu}M$ BA, $2.32{\mu}M$ Kin, and $0.98{\mu}M$ indole-3-butyric acid (IBA). Sub-culturing of shoots every 3 weeks on fresh multiplication medium yielded a consistent proliferation rate of 3.5-fold. Shoot clusters containing three to five shoots were successfully rooted with 100% success on half-strength MS liquid medium supplemented with $9.8{\mu}M$ IBA, $2.85{\mu}M$ indole-3-acetic acid (IAA), $2.68{\mu}M$ naphthaleneacetic acid (NAA), and 3% sucrose. Plantlets grown in vitro were acclimatized and subsequently transferred to the field. Inter-simple sequence repeat analysis has confirmed the genetic uniformity of the tissue-cultured plants up to 27 passages.

High-frequency Plant Regeneration from Cultured Flower Bud Receptacles of Allium hookeri L.

  • Koo, Ja Choon
    • Horticultural Science & Technology
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    • v.32 no.5
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    • pp.694-701
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    • 2014
  • Allium hookeri L. (Alliaceae family) is an important ethnomedicinal plant native to the Himalayan region of Asia. The aim of this research was to establish a high-frequency plant regeneration system for in vitro propagation of A. hookeri. Among the tissue types examined, receptacle explants derived from immature flower buds showed the highest regeneration rate of shoots ($93.33{\pm}4.63%$), roots ($76.67{\pm}7.85%$), and calli ($80.00{\pm}7.43%$) when cultured on Gamborg B5 (B5) medium containing $10{\mu}M$ 6-benzylaminopurine (BA) + $1{\mu}M$ naphthalene acetic acid (NAA), $0.5{\mu}M$ BA + $5{\mu}M$ NAA, and $1{\mu}M$ BA + $10{\mu}M$ NAA, respectively. Shoot multiplication was superior when cultured in liquid rather than on solid medium and relatively high concentrations of BA, ranging from 5 to $10{\mu}M$. Efficient bulblet formation following root induction from shoot clumps was achieved with culture in liquid B5 medium containing 7% (w/v) sucrose. Regenerated bulblets were successfully acclimatized to ex vitro conditions with a greater than 95% survival rate. By this method, a maximum of 62 plantlets per receptacle could be propagated within 9 weeks of initial culture. The in vitro propagation system established in this study will promote A. hookeri biotechnology, including large-scale production of healthy and aseptic clones, preserving parental genotypes with desirable traits, and genetic manipulation to enhance medicinal value.

Effects of Tongdosan on the Cardiovascular System in the Experimental Animals (통도산(通導散)이 실험동물(實驗動物)의 심혈관계(心血管系)에 미치는 영향(影響))

  • Lee, In;Shin, Sun-Ho;Moon, Byung-Soon
    • The Journal of Internal Korean Medicine
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    • v.17 no.1
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    • pp.34-50
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    • 1996
  • The present experiment was desinged to investigate the effects of Tongdosan water extracts on the Cardiovascular System in the Experimental Animals. Thus, the changes of blood pressure and heart rate were measured after oral admini- stration. Measurment of Mortality rate was observed for measuring the effect of Tongdosan water extract. Tongdosan water extract against pulmonary thrombo- embolism induced by collagen the mixture(0.1ml/10g, 2mg/kg B.W) plus serotonin(5mg/kg B.W) in mouse. The effect of Tongdosan water extract was examined by observing the change of collagen-induced platelet aggregation, coagulation activity, ex vivo and in vitro fibrinolytic activity of euglobulin fraction in rats. The results were summarized as followings. 1. Tongdosan dropped the blood pressure in spontaneous hypertensive rat. 2. The drug increased the auricular blood flow in rabbit. 3. The drug relaxed the artery contraction by pretreated norepinephrine in rat. 4. The drug inhibited the death rate of mouse which was led to thromboembo- lism by serotonin and collagen. 5. The drug inhibited the platelet aggregation in rat. 6. The drug prolonged the prothrombin time and activated partial thromboplastin time on the test of plasma coagulation factor activity in rat, but was not valuable. 7. The drug reduced the fibrinogen lyses time of rat ex vivo assay and lyses area was increased. 8. Tongdosan reduced fibrinogen lyses time of rat in vitro assay. According to the above mentioned results, Tongdosan increased the blood flow and dropped the blood pressure by the dilation of blood vessel. And the drug presented the antithrombin acivity, inhibited the platelet aggregation.

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In vitro regeneration of Lycaste aromatica (Graham ex Hook) Lindl. (Orchidaceae) from pseudobulb sections

  • Mata-Rosas, Martin;Baltazar-Garcia, Rosario J.;Moon, Pamela;Hietz, Peter;Luna-Monterrojo, Victor E.
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.157-163
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    • 2010
  • A protocol for in vitro propagation from pseudobulb sections of Lycaste armomatica (Graham ex Hook) Lindl., an ornamental and fragrant orchid, was developed. The effect of four cytokinins: kinetin (K), metatopolin (mT), $N^6$-benzyladenine (BA), and thidiazuron (TDZ), in equimolar concentrations, was investigated. Shoot formation from apical and basal pseudobulb sections was obtained in all treatments. A few medial sections cultured in media supplemented with BA formed protocorm-like bodies. Shoot formation was greater from the basal section than the apical, and mainly occurred in explants cultured in media containing TDZ. The highest average numbers of shoots per explant were achieved from basal sections cultured in media supplemented with TDZ at 4.4, 8.87 and 2.2 ${\mu}M$, forming on average 9.9, 8.6 and 7.3 shoots per explant, respectively. Since the medial pseudobulb section was the worst explant for propagation of L. aromatica, we recommend that pseudobulbs be divided into two sections; the basal half should be cultured in MS medium supplemented with TDZ at 4.4 ${\mu}M$ and the apical half with TDZ at 2.2 ${\mu}M$. Subculturing individual shoots in MS medium without plant growth regulators allows further development and rooting. A survival rate of more than 90% under greenhouse conditions was achieved. This research represents a direct contribution to the conservation and sustainable use of this valuable natural resource.