Optimal conditions for the isolation and culture of protoplasts from a cell suspension culture of an isoleucine–valine-requiring auxotroph of Datura innoxia

1987 ◽  
Vol 65 (8) ◽  
pp. 1736-1740 ◽  
Author(s):  
Praveen K. Saxena ◽  
John King
Keyword(s):  
Sodium Chloride ◽  
Cell Suspension ◽  
Cell Walls ◽  
Cell Suspension Cultures ◽  
Datura Innoxia ◽  
Optimal Conditions ◽  
Murashige And Skoog Medium ◽  
A Cell ◽  
High Yields ◽  
Cell Colony

Conditions have been standardized for obtaining high yields of viable protoplasts from cell suspension cultures of an isoleucine–valine-requiring auxotroph (IV-1) of Datura innoxia P. Mill. Isolation of protoplasts critically required the use of a salt solution, containing sodium chloride and potassium chloride (0.125 M each), as osmotic stabilizers. The yield, viability, and divisional activity of the protoplasts isolated with mannitol or sucrose were poor. Protoplast-releasing enzymes used to isolate IV-1 protoplasts could be used twice without any loss in the yield or viability of the protoplasts. Cultured protoplasts developed cell walls and underwent sustained divisions in a modified Murashige and Skoog medium enriched with organic acids. Pretreatment of isolated protoplasts with glycine (0.1 M) and calcium chloride (0.05 M) prior to culture increased the frequency of cell colony formation. Protoplast-derived cells developed calli on transfer to agar-solidified medium.

Factors Affecting Protoplast Isolation and the Regeneration of Shoot-Like Structures From Protoplast-Derived Callus of Sugarcane (Saccharum spp Hybrids)

1992 ◽  
Vol 40 (6) ◽  
pp. 863 ◽  
Author(s):  
PWJ Taylor ◽  
HL Ko ◽  
SW Adkins
Keyword(s):  
Cell Suspension ◽  
Cell Walls ◽  
Cell Suspension Cultures ◽  
Protoplast Isolation ◽  
Suspension Cultures ◽  
Ms Medium ◽  
Factors Affecting ◽  
Nodular Callus ◽  
Homogeneous Cell ◽  
High Yields

High yields of protoplasts were isolated from non-regenerable, homogeneous cell suspension cultures of sugarcane, compared with regenerable, heterogeneous cell suspension cultures after incubation in an enzyme composition containing Cellulase RS, Pectinase, Macerozyme and Driselase. Higher yields of protoplasts were released from heterogeneous cell suspension cultures after the addition of 10 mg L-1 silver nitrate to the culture medium; however, ethylene production was not involved in protoplast isolation. Use of 0-05-2% Pectolyase Y23 pectinase rather than other pectinases resulted in higher yields of protoplasts from heterogeneous cell suspension cultures. These results suggest that there are differences in the cell walls between cells from heterogeneous and homogeneous cell suspension cultures which affect the isolation of protoplasts. Protoplasts isolated from heterogeneous cell suspension cultures failed to develop beyond the cell division stage. Protoplasts isolated from homogeneous cell suspension cultures and cultured in agarose droplets bathed in modified 8p medium, reformed cell walls, divided and developed into microcallus. Microcallus transferred to solid modified MS medium containing 1 mg -1 .2,4-D developed into callus. Protoplast-derived callus from one cultivar formed compact nodular callus when subcultured onto the same medium containing 1% activated charcoal. Incubation of this callus on MS medium containing BAP at 0.5 mg L-1, then a combination of BAP and fluridone each at 0.5 mg L-1, resulted in the regeneration of small, chlorophyll-containing shoot-like structures. As yet no intact plants have developed from these shoot-like structures.

Potential of Datura innoxia cell suspension cultures for glucosylating hydroquinone

1987 ◽  
Vol 6 (4) ◽  
pp. 275-278 ◽  
Author(s):  
Toshiyuki Suzuki ◽  
Toshihiro Yoshioka ◽  
Mamoru Tabata ◽  
Yasuhiro Fujita
Keyword(s):  
Cell Suspension ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Datura Innoxia

Extracellular polysaccharide from cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender)

1972 ◽  
Vol 50 (10) ◽  
pp. 2031-2037 ◽  
Author(s):  
Deng-Fong Liau ◽  
W. G. Boll
Keyword(s):  
Cell Suspension ◽  
Higher Plants ◽  
Cell Suspension Cultures ◽  
Extracellular Polysaccharide ◽  
Cell Number ◽  
Suspension Cultures ◽  
Logarithmic Phase ◽  
Bush Bean ◽  
Culture Cycle ◽  
High Yields

High yields of extracellular polysaccharide were obtained from cell suspension cultures of root, hypocotyl, and cotyledon of bush bean. Hydrolysates of the three polysaccharide samples contained the same sugars: galacturonic acid, galactose, glucose, mannose, arabinose, and xylose. The relative amounts of the six sugars were not the same in the hydrolysates from the three sources. The extracellular polysaccharide was produced at all times during the culture cycle. Semilogarithmic plots of increase in cell number, and production of extracellular polysaccharide, indicate that production per cell decreased during the logarithmic phase, and increased at the onset of the stationary phase. Production of extracellular polysaccharide, per culture and per cell, was much higher than that reported for other cell cultures of higher plants.

Effect of 5-Azacytidine on the Formation of Secondary Metabolites in Catharanthus roseus Cell Suspension Cultures

1985 ◽  
Vol 40 (1-2) ◽  
pp. 21-25 ◽  
Author(s):  
H.-A. Arfmann ◽  
W. Kohl ◽  
V. Wray
Keyword(s):  
Cell Suspension ◽  
Catharanthus Roseus ◽  
High Performance ◽  
Cell Suspension Cultures ◽  
Elution Profile ◽  
Human Cell Lines ◽  
Production Medium ◽  
Freeze Dried ◽  
A Cell ◽  
C Nmr

A cell suspension culture of Catharanthus roseus was treated during the growth phase with the nucleoside analogue 5-azacytidine, a powerful inducer of cell differentiation, as evidenced with animal and human cell lines. After induction the cell culture was further incubated in a production medium in the absence of the inducer for 10 days. The extraction of the freeze dried cells and analysis by high performance liquid chromatography showed an additional intense peak in the elution profile, not present in extracts of untreated cells. The structure of the newly synthesized metabolite, elucidated by mass spectrometry and 1H and 13C NMR, was a lignan type compound, lirioresinol B mono-β-D-glucoside. This compound has hitherto not been found in Catharanthus roseus plants or cell cultures.

Accumulation of a Novel Red Pigment in Cell Suspension Cultures of Floral Meristem Tissues from Carthamus tinctorius L.

1988 ◽  
Vol 43 (11-12) ◽  
pp. 862-870 ◽  
Author(s):  
Koshi Saito ◽  
Etuko Daimon ◽  
Kazuhito Kusaka ◽  
Sachio Wakayama ◽  
Yoshihiro Sekino
Keyword(s):  
Cell Suspension ◽  
Culture Media ◽  
Cell Suspension Cultures ◽  
Carthamus Tinctorius ◽  
Culture Conditions ◽  
Floral Meristem ◽  
Red Pigment ◽  
A Cell ◽  
Micro Elements ◽  
Β Carotene

Abstract A cell strain originally derived from floral meristem tissues of Carthamus tinctorius (dyer’s saffron) produced substantial amounts of a novel red pigment under controlled culture conditions. The pigment isolated f r om alcoholic extracts of C. tinctorius cultures was compared with authentic carthamin, anthocyanins, betain, and carotenoids. It differed markedly from carthamin and showed none of the characteristic properties of the glycoside or chloride forms of authentic delphinidin, cyanidin, and pelargonidin. Analytic data indicated that this pigment also differs from betanin and from α- and β-carotene. The name “Kurenamin” was tentatively given to this newly isolated red pigment. Effect of the culture media, micro-elements, macro-elements, and putative substrates on the kurenamin production were investigated during cell suspension culture.

scholarly journals Cell Culture of Bursera Linanoe in a Stirred Tank Bioreactor for Production of Linalool and Linalyl Acetate

2017 ◽  
Vol 12 (3) ◽  
pp. 1934578X1701200
Author(s):  
Leticia Pavón-Reyes ◽  
Silvia Evangelista-Lozano ◽  
Gabriela Sepúlveda-Jiménez ◽  
Víctor Chávez Ávila ◽  
Mario Rodríguez-Monroy
Keyword(s):  
Cell Viability ◽  
Cell Suspension ◽  
Cell Suspension Cultures ◽  
Stirred Tank ◽  
Phase Cell ◽  
Naphthalene Acetic Acid ◽  
Linalyl Acetate ◽  
Stirred Tank Bioreactor ◽  
Maximum Biomass ◽  
Murashige And Skoog Medium

Bursera linanoe cell suspension cultures were initiated from callus grown in Murashige and Skoog medium supplemented with naphthalene acetic acid (3.0 mg L−1) and 6-benzylaminopurine (0.5 mg L−1). In flasks, B. linanoe cell cultures grew over a 9 day period, reaching a maximum biomass of 11.16 g DW L−1. Throughout the growth phase, cell viability was constant at 60 – 70%. In contrast, B. linanoe cells growing in a bioreactor achieved a maximum biomass of 22.26 g DW L−1 (after 7 days), and cell viability was constant at 75 - 85%. Production of linalool and linalyl acetate in the bioreactor (3.02 and 2.40 mg g−1 DW, respectively) was significantly greater than that achieved from cells in flask cultures (1.05 and 0.97 mg g−1 DW, respectively). B. linanoe cell suspension culture has potential as an alternative method for the production of essential oils.

Elicitor Induction of Cytochrome P-450 Monooxygenases in Cell Suspension Cultures of Chickpea (Cicer arietinum L.) and Their Involvement in Pterocarpan Phytoalexin Biosynthesis

1991 ◽  
Vol 46 (1-2) ◽  
pp. 58-66 ◽  
Author(s):  
Werner Gunia ◽  
Walter Hinderer ◽  
Uta Wittkampf ◽  
Wolfgang Barz
Keyword(s):  
Cell Culture ◽  
Cell Suspension ◽  
Cicer Arietinum ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Resistant Cultivar ◽  
Susceptible Cultivar ◽  
A Cell ◽  
Cell Culture Line ◽  
Cytochrome P 450

Abstract A yeast glucan elicitor causes the accumulation of the pterocarpan phytoalexins medicarpin and maackiain in chickpea (Cicer arietinum) cell suspension cultures established from seeds. A cell culture line from a chickpea cultivar resistant against its main fungal pathogen Ascochyta rabiei accumulates large amounts (944 nm ol/g fr. wt.) whereas a cell culture line from a susceptible cultivar accumulates only low amounts (38 nm ol/g fr. wt.) of the phytoalexins. This is consistent with differential accumulation of pterocarpan phytoalexins in intact plants [1], The first reactions in the pterocarpan-specific branch of biosynthesis are hydroxylation of the isoflavone intermediate form ononetin in position 2′ or 3′, catalyzed by microsomal cytochrome P-450 monooxygenases. Upon elicitation form ononetin 2′-hydroxylase undergoes a strong transient induction in the cell suspension culture of the resistant cultivar, whereas in the cell culture from the susceptible cultivar it is only slightly induced. In both cell suspension cul­tures the induction of cinnamic acid 4-hydroxylase and of form ononetin 3′-hydroxylase does not show a clear correlation with phytoalexin accumulation. Experiments with different elici­tor concentrations confirm that formononetin 2′-hydroxylase is much more induced in cell cul­tures from the resistant cultivar than from the susceptible one. It is concluded that the massive difference in phytoalexin accumulation between cell suspension cultures from the resistant and susceptible cultivar is determined mainly by the differential induction of form ononetin 2′-hydroxylase activity.

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