• Title/Summary/Keyword: solid culture medium

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Effect of Phenylacetic Acid (PAA) on Embryo Formation in Anther and Microspore Culture of Paeonia lactiflora (작약의 약 및 소포자 배양에서 Phenylacetic Acid [PAA]가 배형성에 미치는 영향)

  • Kwon, Yong-Sham;Shin, Young-Ae;Sohn, Jae-Keun
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.193-198
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    • 2002
  • The objective of this study was to determine the effects of phenylacetic acid (PAA) on embryo production in anther and microspore culture of herbaceous peony (Paeonia lactiflora Pall.). The anthers of herbaceous peony were cultured on MS medium with 0 to 100 mg/L PAA according to two-step culture method. The ruptured anthers were transferred onto embryo formation medium without growth regulators. The MS medium with 2 mg/L PAA was effective in enhancing of direct embryogenesis and producing of normal embryo with two cotyledons from the cultured anthers. However, the increase of PAA concentration more than 5 mg/L PAA inhibited the embryo formation and promoted to callus formation from the anthers. The PAA affects significantly on the division of microspore and embryo formation in shed pollen culture and the best result was obtained from a medium supplement with 2 mg/L PAA. The preculture of anther for 10 days on solid medium with 2 mg/L PAA was effective for embryo formation from shed microspore of herbaceous peony.

Several Factors on Bulblets Regeneration from Callus Culture in Lilium longiflorum 'Celia' (백합 'Gelia' 캘러스로부터 자구 재분화에 미치는 제요인)

  • 박소영;김시동;신세균;이철희;백기엽
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.183-188
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    • 1997
  • Callus from scale segments of Lilium longiflorum 'Gelia' was effectively induced and maintained from unorganized tissue on the semi-solid medium by 0.42% Bacto agar with MS basal salts and vitamins of SH medium supplemented with 0.5 mg/L 2, 4-D, 1.0 mg/L NAA, 0.3 mg/L BA, and 3% sucrose. More than 5% of high sucrose level had inhibiting effect on regeneration capacity of formed callus and decreased callus growth. Various combinations of nitrogen did not effective to proliferate the ELC (Embryogenic-like callus), but friability of callus was increased in the medium containing only nitrate as nitrogen source. 5 mL conditioned medium into 30 mL fresh medium was good for cell growth. However friable cell aggregates during suspension culture had to form hard callus which hindered to establish suspention culture system. Addition of 2 g/L casein hydrolysate increased callus growth and friability of the hard callus. As a result of anatomical observation of callus, organogenesis such as shoots, roots and bulblets was independently induced from callus tissue. Somatic embryogenesis from callus tissue could be observed with low frequency.

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The Effects of Auxin and Casein Hydrolysate on the Growth of Ginseng Hairy Root (인삼모상근의 생장에 미치는 Auxin과 Casein Hydrolysate의 영향)

  • 오승용;박효진;민병훈;양계진;양덕춘
    • Journal of Ginseng Research
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    • v.24 no.3
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    • pp.123-127
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    • 2000
  • In this study, the effects of auxin and casein hydrolysate (CH) on the growth of ginseng hairy root was elucidated. Ginseng hairy root was cultured under light and dark conditions in MS solid and liquid medium with various concentrations of auxin and CH for fifty days. After harvesting the cultures, the fresh and dry weight of cultures were examined, respectively. In the MS solid culture, 1 mg/L of IBA was most effective on the growth of ginseng hairy root under the dark condition, whereas IAA and CH did not affect on the growth of ginseng hairy root. In the MS liquid culture, the growth was maintained regularly by the treatments of IAA and NAA. IBA and CH restrained the growth of ginseng hairy root.

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Callus Induction and Increase in Anti-Inflammatory Activity by Treatment of Methyl Jasmonate in Adenium obesum (석화의 캘러스 유도 및 메틸 자스모네이트 처리에 의한 항염증 활성 증진)

  • Lee, Da Young;Min, Jin Woo;Joo, Gwang Sik;Kang, Hee Cheol
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.2
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    • pp.95-101
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    • 2017
  • Background: Callus cultivation has the advantage of producing a large amount of tissue of a plant in a laboratory regardless of the environment, for extracting an active substance. In the present study, callus formation was induced in the leaves of the succulent plant Adenium obesum (Forssk.) Roem & Schult. After callus cultivation, anti-inflammatory activity tests were conducted, because leaves and stems of A. obesum have been reported to possess biological activity. Methods and Results: In order to induce callus formation, various concentrations of plant growth factors, such as kinetin, naphtha-leneacetic acid (NAA), 6-benzyladenine (BA), and indole-3-acetic acid (IAA) were added to MS solid medium. The maximum callus proliferation was induced by mixed medium consisting of NAA ($2mg/{\ell}$) and BA ($1mg/{\ell}$). In addition, an elicitor was added to the medium under optimal conditions for initiating suspension culture. After suspension culturing, the activities of the callus extracts were compared and analyzed. The cytotoxicity and anti-inflammatory activity tests revealed that the anti-inflammatory activity of the callus extract and the content of phenolic compounds were elevated after treatment of the callus culture with the elicitior. Conclusions: A. obesum callus might be considered as potential source of biologically active anti-inflammatory material.

Comparative Evaluation of Three Culture Methods for the Isolation of Mycobacteria from Clinical Samples

  • Sorlozano, Antonio;Soria, Isabel;Roman, Juan;Huertas, Pilar;Soto, Maria Jose;Piedrola, Gonzalo;Gutierrez, Jose
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1259-1264
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    • 2009
  • We assessed the capacity of two liquid-medium culture methods with automated incubation and reading systems (MB/BacT ALERT 3D System and BACTEC MGIT 960 System) and one solid-medium culture method ($L\ddot{o}wenstein$-Jensen) to detect mycobacteria in different types of clinical samples. Out of 1,770 cultured clinical samples (1,519 of respiratory origin and 251 of non respiratory origin), mycobacteria were isolated in 156 samples (135 M. tuberculosis complex, 8 M. chelonae, 6 M. kansasii, 4 M. fortuitum, 2 M. gordonae, and 1 M. marinum) by at least one of the methods used. The BACTEC MGIT 960 System proved to be the most sensitive method (86.5%), especially in the detection of M. tuberculosis complex (89.1%). However, $L\ddot{o}wenstein$-Jensen culture was the most sensitive (76.2%) to detect nontuberculous mycobacteria. The BACTEC MGIT 960 System showed the lowest mean detection time for mycobacterial growth (15.3 days), significantly shorter than the other two methods. Highest sensitivity (95.5%) and specificity (99.6%) values were obtained using the BACTEC MGIT 960 System with the $L\ddot{o}wenstein$-Jensen culture method, which was also the only combination capable of detecting 100% of the nontuberculous mycobacteria.

Plant Regeneration from Adventitious Roots of Rehmannia glutinosa Liboschitz and Bioreactor Culture (지황 부정근을 이용한 식물체 재분화 및 생물반응기 배양)

  • Jeong, Jae-Hun;Yu, Kee-Won;Kim, Sun-Ja;Choi, Yong-Eui;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.55-60
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    • 2004
  • This experiment was carried out to develop rapid mass propagation via shoot organogenesis system from adventitious roots of Rehmannia glutinosa. The induction of adventitious roots from leaf explants was most favorable to MS solid medium supplemented with 2mg/L IBA. However, the growth of adventitious roots was highest when they were cultured on 1/3 strength MS liquid medium supplemented with 2mg/L IBA. When the adventitious roots were grown in 10L bioreactor, 10g roots as initial inoculum was increased to 225g after 6 weeks of culture. The harvested roots were cultured onto solid medium to induce plant regeneration. The optimal adventitious shoot formation was observed on MS medium supplemented with 2mg/L BA. Rooting of individual shoots was induced after transfer to half strength MS medium without growth regulators. Plantlets after acclimatization were successfully transplanted in the field and no phenotypic variation was observed among them.

Plant Regeneration via Multiple Shoots Formation from Sucker Explants of Rubus fruticosus L. (블랙베리(Rubus fruticosus L.)의 맹아절편체로부터 다경유도를 통한 식물체 재분화)

  • Shin Jeong-Sun;Sim Ock-Kyeong;Lee Jong-Chon;Cho Han-Jik;Kim Ee-Yup;Lee Kang-Seop
    • Korean Journal of Plant Resources
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    • v.18 no.3
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    • pp.456-461
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    • 2005
  • This study was carried out to induce plant regeneration via shoot formation from sucker explants of Rubus fruticosus L. To induce adventitious shoots, sucker explants were sterilized in $1.2\%$ NaOCl solution, and cultured on the MS solid medium supplemented with kinetin (0.5, 1.0, 3.0 mg/L) and BA (0.5, 1.0, 3.0 mg/L), respectively. As above, to induce adventitious shoots, sucker explants were cultured on the MS solid medium supplemented with IBA (0, 0.1, 1.0 mg/L) and BA (0, 0.1, 1.0, 2.0 mg/L). After 4 weeks of culture, the highest frquency $(100\%)$ of shoot formation from sucker explants was obtained from the medium with 1.0 mg/L BA. The highest shoot number per explant from in vitro shoot explants was 5.3. After 10 weeks of culture, the number of shoot per explant was increased. The highest frequency $(85\%)$ of root formation was obtained at 0.5 mg/L glycine medium, when the explant with shoot were cultured on the MS medium containing glycine at various concentrations from 0 to 2.0 mg/L. The survival rate of the plantlets after transfer to plastic pots containing sand, soil, and vermiculite (1:1:1, vol.) was $95\%$. The results indicate that micropropagation procedure can be applied for an efficient mass propagation of Rubus fruticosus.

Cultural Characteristics of Ectomycorrhizal Mushrooms

  • Jeon, Sung-Min;Ka, Kang-Hyeon
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.16-16
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    • 2015
  • Ectomycorrhizal (ECM) mushrooms play a major role in plant growth promotion through symbiotic association with roots of forest trees. They also provide an economically important food resource to us and therefore they have been studied for their artificial cultivation for decades in Korea. We have secured bio-resources of ECM mushrooms from Korean forests and performed their physiological studies. To investigate the cultural characteristics, the fungi were cultured under different conditions (medium, temperature, pH of the medium, inorganic nitrogen source). More than 90% of total 160 strains grew on three solid media (potato dextrose agar, PDA; sabouraud dextrose agar, SDA; modified Melin-Norkrans medium, MMN). The rate of mycelial growth on malt extract agar (MEA) was lower than those of three media (PDA, SDA, MMN). None of the Tricholomataceae strains grew on MEA. Many strains of ECM mushrooms were able to grow at the temperature range of $15{\sim}25^{\circ}C$ on PDA, while they showed poor growth at $10^{\circ}C$ or $30^{\circ}C$. In particular, the growth rates of both Gomphaceae and Tricholomataceae were significantly lower at $10^{\circ}C$ than at $30^{\circ}C$. The optimal pH of many strains was pH 5.0 when they cultured in potato dextrose broth (PDB). Fifty-seven percent of tested strains grew well on medium containing ammonium source than nitrate source. Many strains of Tricholomataceae showed a notable growth on ammonium medium than nitrate medium. Twenty-three percent of strains preferred nitrate source than ammonium source for their mycelial growth. The production and activity of two enzymes (cellulase and laccase) by ECM fungi were also assayed on the enzyme screening media containing CMC or ABTS. Each strains exhibited different levels of enzymatic activities as well as enzyme production. The number of laccase-producing strains was less than that of cellulase-producing strains. We found that 77% of tested strains produced both cellulase and laccase, whereas 2% of strains did not produce any enzymes. The morphological characteristics of mycelial colony were also examined on four different solid media. Yellow was a dominant color in mycelial colony and followed by white and brown on all culture media. ECM mushrooms formed mycelial colonies with a single or multiple colors within a culture medium depending on the strains and culture media. The most common shape of mycelial colony was a circular form on all media tested. Other families except for Amanitaceae formed an irregular colony on MMN than PDA. All strains of Tricholomataceae did not form a filamentous colony on all media. The pigmentation of culture media by mycelial colonies was observed in more than 50% of strains tested on both PDA and SDA. The degree of pigmentation on PDA or SDA was higher than MMN and brown color was dominant than yellow color. The production of exudates from mycelial colony was higher on PDA than MMN. Brown exudates were mainly produced by many strains on PDA or SDA, whereas transparent exudates were mainly produced by strains on MMN. We observed the mycelial colonies with a single or multiple textures in just one culture plate. Wrinkled or uneven colony surfaces were remarkably observed in many strains on PDA or SDA, while an even colony surface was observed in many strains on MMN. Sixty percent of Tricholomaceae strains formed wrinkled surface on PDA. However, they did not form any wrinkle on MMN plate. Cottony texture was observed in mycelia colonies of many strains. Velvety texture was often observed in the mycelial colonies on SDA than PDA and accounted for 60% of Suillaceae strains on SDA.

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Optimized Germination Conditions and Human p53 Expression of Rice Embryo (쌀눈 발아의 최적조건 확립 및 p53 항암 유전자의 발현)

  • Pih, Kyung-Tae;Choi, Ju-Youn;Kim, Keun-Cheol
    • Journal of Life Science
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    • v.25 no.2
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    • pp.158-163
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    • 2015
  • Rice embryo is more abundant than endosperms in nutrients such as proteins, lipids, and vitamin B1. In this study, we constructed p53 plasmid that could be expressed in a plant system, and investigated optimal germination conditions in a variety of media. For construction of p53 plasmid, we performed p53 amplification from pCDNA-p53, subcloned to TA cloning vector, and then reconstructed into pGEM-CaMV plant expression vector. On the other hand, we prepared a variety of imbibition buffers and complete media for efficient germination of the rice embryo. Imbibition buffers prepared with different concentrations of salt or detergent showed no significant effect on germination efficiency. We prepared further culture media, such as solid agar, liquid media, and paper towel to establish the optimal conditions. Rice embryo showed germination rates of more than 70% in the solid medium, more than 60% in the paper towel medium, but less than 25% in liquid media, although germination rate did not differ with varying concentrations of salt and sucrose in culture media. Under the optimal germination conditions, we introduced the p53 plasmid using imbibition method, and finally detected human p53 gene expression in the germinated rice embryo. This method might present a novel, practical approach for evaluating efficient gene expression utilizing imbibition method in rice embryo.

Studies on the production of liquid spawn of Pleurotus ostreatus (Pleurotus ostreatus의 액체 종균 생산에 관한 연구)

  • Kang, Tae-Su;Chun, Byong-IK
    • Journal of Industrial Technology
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    • v.8
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    • pp.13-21
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    • 1988
  • For the improvement of productivity of Pleurotus ostreatus, the production of liquid spawn was studied. The highest liquid spawn production was obtained after shaking culture for 4 days in the culture medium containing 5%(W/V) wheat flour, 0.2%(W/V) yeast extract, 0.1%(W/V)$KNO_3$ 0.05% (W/V) $MgSO_4{\cdot}7H_2O$, 0.05%(W/V) $KH_2PO_4$. The optimum pH and temperature was 7.0 ana $30^{\circ}C$. The period required to complete the mycelial growth after spawning were 28, 22, 10 and 9 days, respectively, when the 2%(V/V) of solid spawn and 2%(V/V), 5% (V/V) and 10%(V/V) of liquid spawn were inoculated. The days required from spawning to fruiting bodies were 38, 34, 28 and 27 days.

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