• Title/Summary/Keyword: Solid culture

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Somatic Embryogenesis and Plant Regeneration from Stem Tissues of Orostachys japonicus A. Berger

  • Song, Min-Jung;Park, Young-Goo
    • Journal of Plant Biotechnology
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    • v.34 no.3
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    • pp.181-187
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    • 2007
  • Orostachys japonicus A. Berger is a Perennial herbaceous plant which has been traditionally used as an anti-inflammatory agent to treat hepatitis and as an anticancer agent. The objective of this study was 1) to establish and proliferate in vitro plant of O. japonicus 2) to induce indirect somatic embryogenesis from O. japonicus. General calli and embryogenic calli in all ranges of 2,4-D and BA combination, were induced and were best at 22% (embryogenic cell) in 5.0 mg/L 2,4-D and 0.5 mg/L BA combination. Embryogenic cell line was maintained by subculture at 2 week intervals and transferred to solid and liquid medium for embryo formation. In solid medium culture, globular and heart shaped embryos were observed in MS medium containing 5.0 mg/L 2,4-D and 0.5 mg/L BA combination. The number of embryos was 6.5 per 0.5 g cell, and then the immature embryos transferred to MS basal medium for embryo development. In a suspension culture of embryogenic cells, globular and heart shaped embryos were emerged in MS medium supplemented with 3.0 mg/L 2,4-D and 0.3 mg/L BA combination after 10 days of incubation. The embryo formation rate was about 33% by suspension culture. The ratio of embryo germination was 60.9%, on the other side, the root formation rate was 74.3% in 1/2 MS continuously.

Establishment of Cell Suspension Cultures and Plant Regeneration in White Dandelion (Taraxacum coreanum NAKAI.)

  • Sun, Yan-Lin;Kim, Jae-Hak;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.24 no.3
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    • pp.280-285
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    • 2011
  • In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.

The Optimal Condition for Production of Red Pigment by Monascus anka on Solid Culture (고체배양에 의한 Monascus anka의 적색색소 생성의 최적 조건)

  • 이승민;김현수;유대식
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.2
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    • pp.155-160
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    • 2003
  • The optimum cultural conditions for production of red pigment from Monascus anka KCTC 6121 on solid culture were studied. The optimal conditions were found that the strain was cultivated on polished rice with 25% initial moisture content, at 3$0^{\circ}C$, 90% humidity for 12 days. It was also found that the maximum red pigment was extracted when the final culture was left in 80% ethanol for 2 days. The light stability of the extracted red pigment was relative stable since the discoloration rate was less than 8% in 30 days under the indirect light.

Effects of Temperature and Culture Media Composition on Sporulation, Mycelial Growth, and Antifungal Activity of Isaria javanica pf185

  • Lee, Jang Hoon;Lee, Yong Seong;Kim, Young Cheol
    • Research in Plant Disease
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    • v.27 no.3
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    • pp.99-106
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    • 2021
  • The fungal isolate Isaria javanica pf185 has potential as a mycopesticide because it demonstrates insecticidal activity against the green peach aphid and antifungal activity against Colletotrichum gloeosporioides. For commercialization of this isolate, determination of the optimal and least expensive culture conditions is required; however, these data are not currently available. This study describes the conditions for optimal development of conidia and production of metabolites for the biocontrol of the fungal pathogen. The optimal culture conditions were examined using cultures on solid agar and liquid media. High growth temperature enhanced spore formation but reduced antifungal activity in both solid and liquid media. The highest spore yield was obtained in a medium containing glucose as a carbon source and yeast extract as a nitrogen source. Soybean powder and wheat bran were effective nitrogen sources that promoted spore production and antifungal activity of the isolate. These results revealed the basic, cost-effective growth media for commercial production of a biopesticide with insecticidal and antifungal properties for use in integrated pest management.

Physiological Activity of Roasted Coffee prepared from Fermented Green Coffee Bean with Monascus ruber Mycelium (홍국균(Monascus ruber) 균사체-커피생두 발효물로부터 조제된 원두커피의 생리활성)

  • Kim, Hoon;Suh, Hyung-Joo;Shin, Ji-Young;Hwang, Jong-Hyun;Yu, Kwang-Won
    • The Korean Journal of Food And Nutrition
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    • v.29 no.1
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    • pp.1-11
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    • 2016
  • To enhance the physiological activities of roasted coffee (RC), 30 kinds of green coffee beans (GCB) with different cultivating areas and varieties were fermented with Monascus ruber mycelium (MR) by solid-state culture. After the dried MR-fermented GCB was subjected medium roasting, each RC was extracted with hot-water. Among the hot-water extracts, the highest yield was the hot-water extract of RC from MR-fermented Indonesia Mandheling GCB (15.5%). However, the hot-water extract of RC from MR-fermented Ethiopia Sidamo GCB showed significantly higher polyphenolic contents (3.08 mg GAE/100 mg) and ABTS free radical scavenging activity (25.41 mg AEAC/100 mg). Meanwhile, the hot-water extract of RC from MR-fermented Vietnam Robusta GCB showed not only the effective inhibition of $TNF-{\alpha}$ level (73.7% inhibition of LPS-stimulated control) from LPS-stimulated RAW 264.7 cells but also significant inhibition of lipogenesis (63.5% inhibition of lipid differentiation control) in 3T3-L1 pre-adipose cells. In conclusion, these results suggest that roasted coffees from Ethiopia Sidamo and Vietnam Robusta green coffee beans fermented with Monascus ruber mycelium using solid-state culture could have industrial applications as functional coffee beverages.

Solid Substrate and Submerged Culture Fermentation of Sugar Cane Bagasse for the Production of cellulase and Reducing Sugars by a Local Isolate, Aspergillus terreus SUK-1

  • Wan Mohtar, Yusoff;Massadeh, Muhannad Illayan;Kader, Jalil
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.770-775
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    • 2000
  • Several process parameters were studied to ascertain the effect on degradation of sugar cane bagasse in relation to the production of cellulase enzyme and reducing sugars by Solid Substrate Fermentation (SSF) and Submerged Culture Fermentation (SCF) of Aspergillus terreus SUK-1. The effect of air-flow rate (0-1.3 v/v/m), of different ratios of substrate weight to liquid volume (1:6, 1:10, 1:20, and 1:30 w/v, g/ml), scale-up effect (10, 20, and 100 times of 1:10 ration, w/v) and the effect of temperature (30, 40, 50, and $60^{\circ}C$) in SSF were studied. Air-flow rate of 1.0 v/v/m gave the highest enzyme activity (FPase 0.25 IU/ml, CMCase 1.24 IU/ml) and reducing sugars concentration (0.72 mg/ml). Experiment using 1:10 ratio (w/v) was found to support maximum cellulase activity (FPase 0.58 IU/ml, CMCase 1.97 IU/ml) and reducing sugar concentration (1.23 mg/ml). Scaling-up the ratio of 1:10(w/v) by a factor of 20 gave the highest cellulase activity (FPase 0.71 IU/ml, CMCase 2.25 IU/ml) and reducing sugar concentration (3.67 mg/ml). The optimum temperature for cellulase activity and reducing sugar production was $50^{\circ}C$(FPase 0.792 IU/ml, CMCase 2.25 IU/ml and 3.85 mg/ml for reducing sugar concentration). For SCF, the activity of cellulase enzyme and reducing sugar concentration was found to be lower than that obtained for SSF. The highest cellulase activity obtained in SCF was 50% lower than the highest cellulase activity in SSF, while for reducing sugar concentration, the highest concentration obtained in SCF was 90% lower than that obtained in SSF.

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Influence of the Culture Media and the Organic Matter in the Growth of Paxillus ammoniavirescens (Contu & Dessi)

  • Cagigal, Elena Fernandez-Miranda;Sanchez, Abelardo Casares
    • Mycobiology
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    • v.45 no.3
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    • pp.172-177
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    • 2017
  • The genus Paxillus is characterized by the difficulty of species identification, which results in reproducibility problems, as well as the need for large quantities of fungal inoculum. In particular, studies of Paxillus ammoniavirescens have reported divergent results in the in vitro growth while little is known of its capacity to degrade organic matter. For all the above, and assuming that this variability could be due to an inappropriate culture media, the aim of this study was to analyse growth in different culture media (MMN, MS, and 1/2 MS) and in the case of MMN in presence/absence of two types of organic matter (fresh litter and senescence litter) to probe the saprophytic ability of P. ammoniavirescens. We also evaluated the effects of pH changes in the culture media. Growth kinetics was assessed by weekly quantification of the area of growth in solid culture media over 5 wk, calculating the growth curves and inflection points of each culture media. In addition, final biomass after 5 wk in the different culture media was calculated. Results showed that best culture media are MS and 1/2 MS. Moreover, an improvement in growth in culture media containing decomposing fall litter was observed, leading to confirm differences in the culture media of this species with others of the same genus. Further, we established that all growth media suffered a significant acidification after fungal growth.

Phytotoxicity of Endophytic Fungi and Characterization of a Phytotoxin Isolated from Gliocladium catenulatum from Pinus densiflora (식물내생곰팡이의 제초활성 검정 및 소나무에서 분리한 Gliocladium catenulatum이 생산하는 제초활성 물질의 특성 규명)

  • Choi, Gyung-Ja;Park, Joong-Hyeop;Kim, Heung-Tae;Lee, Seon-Woo;Choi, Jung-Sup;Hong, Kyung-Sik;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Mycology
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    • v.32 no.1
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    • pp.8-15
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    • 2004
  • This study was conducted to discover new phytotoxins which may be used as lead molecules for the development of new herbicides. A total of 187 endophytic fungi were isolated from 11 plant species, which were collected from 8 locations in Korea. Their herbicidal activities were screened in vivo by herbicidal and duckweed bioassays after they were cultured in potato dextrose broth and rice solid media. Both fermentation broth and solid culture extract of Gliocladium catenulatum F0006 isolated from red pine (Pinus densiflora) showed 70% herbicidal activity only against cocklebur (Xanthium strumarium) out of the 10 weeds tested. Solid culture extract of F0034 isolated from arrowroot (Pueraria thunbergiana) exhibited 20 to 100% herbicidal activities against all of the weeds. Especially, shattercane (Sorghum bicolor), barnyardgrass (Echinochloa crus-galli), large crabgrass (Digitaria sanguinalis), and fall pauicum (Panicum dichtomiflorum) were sensitive to the solid culture extract of F0034. In addition, solid culture extract of F0043 isolated from red pine displayed 20% to 70% herbicidal activities only against 5 grass species, but not against 5 broad-leaf plant species. On the other hand, as the results of duckweed assay, 8 fermentation broths showed 100% growth inhibitory activity at concentrations less than 5.0% of culture supernatants and 12 solid cultures had a potent inhibitory activity against duckweed growth. A toxic metabolite was purified from the solid cultures of G. catenulatum F0006 by repeated column chromatography and bioassay. It caused a phytotoxic syndrome only on cocklebur out of the 10 weeds tested; it completely killed cocklebur seedlings at $500\;{\mu}g/ml$ and showed 85% herbicidal activity against cocklebur at $100\;{\mu}g/ml$. The molecular weight of the toxic metabolite is 238 daltons and its structure determination is underway.

Effect of Medium Components and Culture Methods on Shoots Regeneration from Athyrium niponicum (개고사리의 기내 포자체 재생에 미치는 배지구성물질 및 배양방법의 영향)

  • Shin, So-Lim;Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.24 no.2
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    • pp.113-120
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    • 2011
  • Present studies are carried out to find media components and culture methods for in vitro propagation of Athyrium niponicum and to establish the optimal economic masspropagation systems. Among pinnae, petiole and rhizome segments only rhizome segments produced young plants. Rhizome segments showed vigorous plant regeneration on 1/2MS medium and supplement to 1% sucrose and 50 $mg{\cdot}L^{-1}$ $NaH_2PO_4$ were promoted the plant regeneration from rhizome segments. Kinetin was better than BA for plant regeneration and combination with 2 ${\mu}M$ kinetin and 5 ${\mu}M$ IBA was most efficient for plant regeneration. Solid or liquid medium with or without 0.1% qactivated charcoal in modified 1/2MS medium (1% sucrose, 50 $mg{\cdot}L^{-1}$ $NaH_2PO_4$, 2 ${\mu}M$ kinetin, 5 ${\mu}M$ IBA, pH 5.8) were used to find the optimal culture methods. The plant regeneration from rhizome segments were most vigorous on solid medium without activated charcoal. The addition of activated charcoal were inhibited the plant regeneration from rhizome segments not only on solid medium but also liquid stationary or suspension culture.

Isolation and Culture of Protoplasts from Hypocotyl-derived Callus of Soybean (Glycine max) (대두 (Glycine max) 부배유 유래 칼루스의 원형질체 분리 및 배양)

  • 이광웅
    • Journal of Plant Biology
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    • v.28 no.3
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    • pp.233-241
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    • 1985
  • The isolation and culture of protoplasts from hypocotyl-derived calluses of Glycine max (L.) Merr. cv. Jangyeop were obtained by digestion for 6 hrs in an enzyme solution containing 3.5% cellulase, 1.5% macerozyme, 10% sorbitol and 0.1% CaCl2.2H2O at pH 5.8. Newly formed cell wall of protoplasts cultured in MS agar medium containing 10 $\mu$M $\alpha$-naphthaleneacetic acid (NAA) and 32 $\mu$M N6-benzylaminopurine (BAP) could be observed after 24 hrs culture. The first cell division of the protoplasts was observed after 3 days of culture; cell clusters after 2 weeks of culture. When transferred to solid media, the protoplasts formed cell clusters gave rise to proliferating calluses.

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