• Journal of Plant Biotechnology
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• v.4 no.1
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• pp.23-27
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• 2002

To establish an in vitro mass production system of grape anthocyanin pigments through callus and cell suspension culture, the effects of nitrogen source and sucrose on fresh weight and anthocyanin production in cell suspension culture of 'Sheridan' grape level were studied. When the medium was devoid of $NO_3^-$, cell fresh weight was either remained stable (1% sucrose) or slightly decreased with culture time (2,3, and 4% sucrose). When $NH_4^-$ was lacking, 3% sucrose was most favorable for cell growth. When $NH_4^-$ was supplied as N source, the anthocyanin content of 2% sucrose containing medium was maintained 2 times higher than other levels till day 8 in culture, then that of 3 and 4% sucrose which peaked at day 12 thereafter. The anthocyanin content was low than $NO_3^-$-free media. Total anthocyanin content in $NH_4^-$-free medium was just about a half of that of $NH_4^+$ medium. Anthocyanin production of 2% sucrose in $NH_4^+$ medium was maintained about 3-fold till day 8, then decreased thereafter. In $NH_4^+$ medium, pH decreased gradually with final pH of 3.5 to 4.0, while pH in $NH_4^+$-free medium increased with final pH of 6.5 to 7.5.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.135-141
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• 2003

The human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene was introduced into tobacco plants. The cell suspension culture was established from leaf-derived calli of the transgenic tobacco plants in order to express and secrete a biologically active hGM -CSF. The recombinant hGM-CSF from the transgenic plant cell culture (prhGM-CSF) was identified as a yield of about 180 ${\mu}$g/L in the culture filtrate, as determined by ELISA. The addition of 0.5 g/L polyvinylpyrrolidone (PVP) to the plant cell culture medium both stabilized the secreted prhGM-CSF and increased the level of production approximately 1.5-fold to 270 ${\mu}$g/L. The biological activity of the prhGM-CSF was confirmed by measuring the proliferation of the hGM-CSF-dependent cell line, TF-1. Interestingly, the specific activity of the prhGM-CSF was estimated to be approximately 2.7 times higher than that of a commercially available preparation from E. coli.

• Plant Biotechnology Reports
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• v.2 no.1
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• pp.33-39
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• 2008

In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with $0.5mg\;l^{-1}$ 2,4-D and $1.0mg\;l^{-1}$ Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of cell aggregates was implanted on MS agar medium supplemented with Kin ($0.0-2.0mg\;l^{-1}$), $GA_3$ ($0.0-2.0mg\;l^{-1}$) and AS ($80.0mg\;l^{-1}$). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on $GA_3$-free medium, but the best morphological quality of embryos was observed in the presence of $0.5-1.0mg\;l^{-1}$ Kin, $0.5mg\;l^{-1}$ $GA_3$ and $80.0mg\;l^{-1}$ AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension and 100% of uniformity for cotyledon suspension.

• Journal of Plant Biology
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• v.33 no.3
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• pp.189-196
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• 1990

Suspension cell lines have been newly established from the calli derived from the immuature embryo culture of hexapolid (Triticum aestivum var. sicco), tetrapolid (T. durum) and diploid (T. tauchii or Aegilops squarrosa) wheat species. The chromosomal variation in suspension cultured cell lines was examined and old cell line, C82d, established from T. aestivum var. copain was also used. New method using 1-bromonaphthalene for metaphase rapping of suspension cells was developed. Variation in chromosome number was observed among all the suspension lines. Cells with doubled chromosome number and deleted chromosome were also observed. Extensive structural changes in chromosome were found in C82d line. Chromosome aberrations showed loss of chromosome arms and chromosome segment. The mean chromosome number in suspension cells of T. aestivum var. sicco was 40, in C82d line 33, in T. durum 28 and in T. tauchii 14. The stability of chromosome in suspension cells of diploid and tetrapolid wheats was higher than that of hexaploid wheat.

• Journal of Life Science
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• v.12 no.5
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• pp.577-583
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• 2002

The growth kinetics and the production of $\beta$-glucuronidase from transgenic tobacco's suspension culture was investigated in the flask culture and a 2.5 L bubble column reactor. The growth of bubble column reactor was similar to that of flask culture. However, in the bubble column reactor, the production of $\beta$ -glucuronidase reached 2850 U/mg (85-fold higher than that of flask culture). In both case, the production level of $\beta$ -glucuronidase was fluctuated, which was resulted from periodical degradation of the protein. Sucrose is important component in plant culture medium. Twice addition of sucrose in bubble column reactor could not improve cell growth, since other components in a medium were already depleted. However, the addition of sugar decreased cell size, which facilitated the operation of bioreactor. The production of $\beta$ -glucuronidase was continuously increased, however final concentration of $\beta$ -glucuronidase was similar to that without sucrose addition.

• Journal of Plant Biotechnology
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• v.5 no.2
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• pp.131-135
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• 2003

Callus and cell suspension cultures of Eurycoma longifolia Jack were initiated from leaves of different trees. The leaf explant of tree Eu9 produced the most calli and also induced high cell biomass in the cell suspension culture. Optimum production of cell biomass could be initiated in proliferating culture medium with a pH of 5.75 prior to autoclaving. The effects of macronutrient inorganic salts of Murashige and Skoog (MS) liquid medium supplemented with X on production of cell biomass of Eurycoma longifolia were also investigated. The highest cell biomass was produced in MS medium containing macronutrients of $21\;mM\;NH_4NO_3,\;12.25\;mM\;KNO_3,\;3.00\;mM\;CaCl_2.2H_2O,\;0.575\;mM\;MgSO_4.7H_2O$, and $1.83\;mM\;KH_2PO_4$. A new medium labeled as TAM was formulated for the production of Eurycoma longifolia cell biomass in the cell suspension culture.

• Korean Journal of Medicinal Crop Science
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• v.12 no.6
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• pp.494-499
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• 2004

A method for long-term conservation of somatic embryos of Eleutherococcus senticosos was described. Suspension cultured globular somatic embryos were successfully conservated for 36 months at $4^{\circ}C$. The embryos resumed growth within two weeks when returned to MS liquid medium containing $0.2\;mg/{\ell}$. 2,4-dichlorophenoxy acetic acid. The optimal condition for cell proliferation was achieved when somatic embryos cultured at $32^{\circ}C$ in 1/3 MS liquid medium, and about 1.2 g of embryogenic cell was induced from 150 globular embryos after 6 weeks of suspension culture. The embryogenic cells produced from these somatic embryos exhibited normal plant regeneration on auxin-free medium.

• The Plant Pathology Journal
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• v.32 no.3
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• pp.228-241
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• 2016

In our previous study, we reported that a novel endophytic bacterium Bacillus oryzicola YC7007 has suppressed bacterial diseases of rice via induced systemic resistance and antibiotic production. This endophytic strain, B. oryzicola YC7007 was used as a biological control agent against bakanae disease of rice caused by Fusarium fujikuroi, and its mechanism of interaction with the pathogen and the rice was further elucidated. Root drenching with B. oryzicola YC7007 suspension reduced the disease severity of bakanae significantly when compared with the untreated controls. The treatments of B. oryzicola YC7007 suspension ($2.0{\times}10^7cfu/ml$) to the rice rhizosphere reduced bakanae severity by 46-78% in pots and nursery box tests containing autoclaved and non-autoclaved soils. Moreover, in the detached rice leaves bioassay, the development of necrotic lesion and mycelial expansion of F. fujikuroi were inhibited significantly by spraying the culture filtrate of B. oryzicola YC7007. Drenching of ethyl acetate extracts of the culture filtrate to the rhizosphere of rice seedlings also reduced the bakanae disease severity in the plant culture dish tests. With the root drenching of B. oryzicola YC7007 suspension, the accumulation of hydrogen peroxide was observed at an early stage of rice seedlings, and a hormonal defense was elicited with and without pathogen inoculation. Our results showed that the strain B. oryzicola YC7007 had a good biocontrol activity against the bakanae disease of rice by direct inhibition, and was also capable of inducing systemic resistance against the pathogen via primed induction of the jasmonic acid pathway.

• Journal of Plant Biology
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• v.34 no.1
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• pp.19-23
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• 1991

Culture of embryogenic callus and suspension were induced from rice seeds in MS2.5 medium. In hormone free N6 medium, whole plantlets were regenerated from embryogenic callus. We observed cell division and reformation of embryogenic callus on culture of protoplast isolated from embryogenic cell suspensions. In addition, we studied the influencing factors on viability of protoplast treated with electroporation. Viability was decreased according to the increase of voltage and capacitance during electroporation. An optimal level of viability was obtained after treatment with $200-300\;V/1180\;\mu\textrm{F}$ in HEM buffer at $4^{\circ}C$..

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.6
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• pp.331-334
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• 2002

Culture conductivity and on-line NADH fluorescence were used to measure cellular growth in plant cell suspension cultures of Podophyllum hexandrum. An inverse correlation between dry cell weight and medium conductivity was observed during shake flask cultivation. A linear relationship between dry cell weight and culture NADH fluorescence was obtained during the exponential phase of batch cultivation In a bioreactor under the pH stat (pH 6) conditions. It was observed that conductivity measurement were suitable for biomass characterisation under highly dynamic uncontrolled shake flask cultivation conditions. However, if the acid/alkali feeding is done for pH control the conductivity measurement could not be applied. On the other hand the NADH fluorescence measurement allowed online-in situ biomass monitoring of rather heterogenous plant cell suspension cultures in bioreactor even under the most desirable pH stat conditions.