• Journal of Plant Biotechnology
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• v.5 no.2
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• pp.121-129
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• 2003

Callus cultures of Azadirachta indica flower petals were established on MS medium supplemented with naphthalene acetic acid (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$. Cell cultures of Azadirachta indica were established and studied the growth and production kinetics. Half 85 medium supplemented with dicamba (2 mg/L), kinetin (1 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable for initiation and maintenance of cell cultures from the calli. MS medium supplemented with naphthalene acetic acid (NAA) (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable as production medium. Around $80\%\;(0.05\%\;w/v)$ of azadirachtin was found to be intracellular. The effect of various precursors, elicitors, permeabilizing agents and growth retardants in cell cultures was studied. The addition of precursors sodium acetate (10 mg/L), squalene (10 mg/L), isopentenyl pyrophosphate (1 mg/L) and geranyl pyrophosphate (1 mg/L) to the cell cultures on day 3 has shown significant increase in bioproduction of azadirachtin $(64.94{\pm}4.40\;mg/L,\;72.81{\pm}0.04\;mg/L,\;51.63{\pm}1.26\;mg/L\;and\;30.70{\pm}0.28\;mg/L\;respectively)$ over the control cultures $(4.70{\pm}0.27 mg/L)$. $5\%$ v/v cell extracts of Fusarium solani has shown moderate increase in the content of azadirachtin $(5.71{\pm}0.34\;mg/L)$ when compared to control cultures $(2.40{\pm}0.56\;mg/L)$. The addition of methyl jasmonate $(500\;{\mu}M/L)$ on day 3 has shown $\~4$ fold improvement in bioproduction of azadirachtln $(6.92{\pm}0.11\;mg/L)$ when compared to control cultures $(1.63{\pm}0.02\;mg/L)$. There was no significant effect of the studied growth retardants and permeabilizing agents on bioproduction of azadirachtin. Cells are cultivated in large volumes using the effective precursors.

• KSBB Journal
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• v.15 no.4
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• pp.337-341
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• 2000

The decanter is very useful to harvest biomass from plant cell cultures in large-scale process. It is very important to obtain high yield and low moisture content in recovered biomass so as to minimize solvent usage in subsequent extraction steps. Effluent clarity was also affected by the differential speed although this affect was more dramatic at higher flow rates than at lower flow rates. Moisure content was largely unaffected by flow rate. A decrease in moisture content was evident as differential speed decreased.

• Journal of Plant Biotechnology
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• v.6 no.2
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• pp.125-130
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• 2004

Callus and cell suspension cultures of Eurycoma longifolia Jack could be an alternative supply of 9-hydroxycanthin-6-one and 9-methoxycanthin-6-one. The callus tissues were initiated from leaves of different trees. The friable calli were used for the preparation of the cell suspension cultures of E. longifolia. The leaf explant of tree Eu-9 produced the most callus and also induced high cell biomass in the cell suspension culture, but it produced low quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. The leaf explant from tree Eu-8 produced low quantity of callus and cell biomass, but produced the highest quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. Optimum production of cell biomass was obtained on cell culture medium with pH 5.75 prior to autoclaving, but high alkaloids content could be induced in culture medium in acidic condition with pH 4.75 and 5.25 prior to autoclaving.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.9-13
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• 2010

Guggulsterone, a hypolipidemic natural agent, is produced in resin canals of the plant Commiphora wightii. In this study, the stimulatory effects of growth retardants [ALAR (N,N-dimethylaminosuccinamic acid) and CCC (chlormequat chloride)] and fungal elicitor on guggulsterone accumulation in cell cultures of C. wightii are reported. CCC at $1\;mg\;l^{-1}$ enhanced guggulsterone content (${\sim}123\;{\mu}g\;l^{-1})$ when added on the fifth day after inoculation, while ALAR at $2.5\;{\mu}g\;l^{-1}$ increased guggulsterone content (${\sim}116\;{\mu}g\;l^{-1}$) when added on the tenth day. In a two-stage fed-batch process, combined treatment with fungal elicitor and growth retardant caused a significant increase (${\sim}353\;{\mu}g\;l^{-1}$) in guggulsterone content in cell cultures after 17 days of growth. This represents an approximately fivefold increase over the guggulsterone contents in initial cultures of this plant.

• Journal of Plant Biotechnology
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• v.4 no.4
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• pp.143-150
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• 2002

A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.3
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• pp.138-149
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• 2002

Plant cell culture provides a viable alternative over whole plant cultivation for the production of secondary metabolites. In order to successfully cultivate the plant cells at large scale, several engineering parameters such as, cell aggregation, mixing, aeration, and shear sensitivity are taken into account for selection of a suitable bioreactor. The media ingredients, their concentrations and the environmental factors are optimized for maximal synthesis of a desired metabolite. Increased productivity in a bioreactor can be achieved by selection of a proper cultivation strategy (batch, fed-batch, two-stage etc.), feeding of metabolic precursors and extraction of intracellular metabolites. Proper understanding and rigorous analysis of these parameters would pave the way towards the successful commercialization of plant cell bioprocesses.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.04a
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• pp.139-139
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• 2005

• KSBB Journal
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• v.15 no.4
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• pp.398-401
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• 2000

Taxiods inclusive paclitaxel were produced isolated and purified from plant cell cultures of Taxus chinensis in large-scale process. their structures were elucidated by spectroscopic analyses. These compounds were exactly identical as those in previous studies from the other biomasses of Taxus chinensis and also other species. Also the concentrations of these compounds were compared with the concentration of the paclitaxel in various batches of plant cell cultures. As paclitaxel concentration increased at the end of cell cultures. the concentrations of the other paclitaxel derivatives decreased. The profile of these taxoids production can provide information for better understanding of structure-activity relationships and biosynthesis Importantly it can be utilized as an useful parameter for the quality control of paclitaxel production.

• Journal of Microbiology and Biotechnology
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• v.19 no.6
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• pp.573-581
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• 2009

The complex enzyme pool secreted by the phytopathogenic fungus Fusarium graminearum in response to glucose or hop cell wall material as sole carbon sources was analyzed. The biochemical characterization of the enzymes present in the supernatant of fungal cultures in the glucose medium revealed only 5 different glycosyl hydrolase activities; by contrast, when analyzing cultures in the cell wall medium, 17 different activities were detected. This dramatic increase reflects the adaptation of the fungus by the synthesis of enzymes targeting all layers of the cell wall. When the enzymes secreted in the presence of plant cell wall were used to hydrolyze pretreated crude plant material, high levels of monosaccharides were measured with yields approaching 50% of total sugars released by an acid hydrolysis process. This report is the first biochemical characterization of numerous cellulases, hemicellulases, and pectinases secreted by F. graminearum and demonstrates the usefulness of the described protein cocktail for efficient enzymatic degradation of plant cell wall.

• Korean Journal of Plant Tissue Culture
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• v.25 no.6
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• pp.501-505
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• 1998

Conditions for high frequency plant regeneration from cryopreserved embryogenic cell suspension cultures derived from hypocotyl explants of cucumber (Cucumis sativus L.) are described. Cells cryoprotected with a mixture of 2 M DMSO and 0.4 M sucrose exhibited a regeneration frequency of 85%. However, cells cryoprotected with different concentrations of glycerol showed no regeneration after cryopreservation. Pretreatment of cells in a high osmotic medium was not necessary to the process. Upon transfer to MS medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid, regenerated calli gave rise to numerous somatic embryos, then underwent development into plantlets.