Comparison of growth and extracellular polysaccharide of cotyledon cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender) grown in coconut-milk medium and synthetic medium

1975 ◽  
Vol 53 (15) ◽  
pp. 1542-1548 ◽  
Author(s):  
S. Mante ◽  
W. G. Boll
Keyword(s):  
Synthetic Medium ◽  
Cell Suspension Cultures ◽  
Extracellular Polysaccharide ◽  
Suspension Cultures ◽  
Coconut Milk ◽  
Neutral Sugar ◽  
Bush Bean ◽  
Cotyledon Cell ◽  
Neutral Sugars ◽  
Culture Cycle

Growth and extracellular polysaccharide (EP) production by cotyledon suspension cultures of bush bean were compared in coconut-milk (CM) medium and synthetic medium. EP was produced at all stages of the culture cycle investigated. The same neutral sugars were present in the EP recovered from both media and the composition showed some changes during the culture cycle. Factors in coconut milk had relatively little effect on the neutral sugar composition of the EP.

Effect of either 2,4-dichlorophenoxyacetic acid or kinetin on production and composition of various fractions from extracellular polysaccharides produced by cotyledon cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender)

1978 ◽  
Vol 56 (15) ◽  
pp. 1816-1822 ◽  
Author(s):  
S. Mante ◽  
W. G. Boll
Keyword(s):  
Cell Suspension ◽  
Cell Suspension Cultures ◽  
Extracellular Polysaccharide ◽  
Suspension Cultures ◽  
Extracellular Polysaccharides ◽  
Dichlorophenoxyacetic Acid ◽  
Neutral Sugar ◽  
Bush Bean ◽  
Cotyledon Cell ◽  
2,4 Dichlorophenoxyacetic Acid

Cotyledon cell suspension cultures of bush bean required 2, 4-dichlorophenoxyacetic acid (2,4-D) for growth. Kinetin was not essential but was required for optimum growth. Both of the regulators were required for optimum production of extracellular polysaccharide (EP).The two regulators had different effects upon the production of three polysaccharide fractions (two pectins and a neutral polysaccharide) isolated from the EP at various stages of the culture cycle.The neutral sugar composition of the pectin fractions from all treatments, including regulator treatments, showed considerable fluctuation during the culture cycle and could be changed markedly by regulator treatments. Changes in composition of the neutral polysaccharide were slight. As a consequence of these results it is now possible to obtain cells differing in the nature of the systems synthesizing, or controlling the synthesis of, the polysaccharides.

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.

Changes in the amount and composition of fractions from extracellular polysaccharide during the culture cycle of cotyledon cell suspension culture of bush bean (Phaseolus vulgaris cv. Contender)

1976 ◽  
Vol 54 (3-4) ◽  
pp. 198-201 ◽  
Author(s):  
S. Mante ◽  
W. G. Boll
Keyword(s):  
Phaseolus Vulgaris ◽  
Suspension Culture ◽  
Cell Suspension Culture ◽  
Cold Water ◽  
Extracellular Polysaccharide ◽  
Water Soluble ◽  
Bush Bean ◽  
Cotyledon Cell ◽  
Neutral Sugars ◽  
Culture Cycle

Three different polysaccharides (cold-water-soluble pectin, EDTA-soluble pectin, and neutral polysaccharides) were separated from extracellular polysaccharide produced by bush-bean cotyledon suspension cultures. The different polysaccharides were associated with different phases of the culture cycle. The neutral sugars present in the pectin fractions showed considerable fluctuation during the culture cycle as compared with the rather small changes in the levels of the sugars present in the neutral polysaccharides.

Callus and cell suspension culture of bush bean (Phaseolus vulgaris)

1970 ◽  
Vol 48 (6) ◽  
pp. 1119-1130 ◽  
Author(s):  
Deng-Fong Liau ◽  
W. G. Boll
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Cultures ◽  
Dry Weight ◽  
Cell Number ◽  
Callus Cultures ◽  
Suspension Cultures ◽  
Coconut Milk ◽  
Mineral Salts ◽  
Bush Bean

A solid medium was developed for callus cultures originating from explants of root, hypocotyl, and cotyledon of seedlings of bush bean, and a liquid medium was developed for the growth of cell suspension cultures derived from the callus cultures. Some unsatisfactory media are recorded. Concentrations of mineral salts for cell suspension cultures are lower than for callus cultures. Both coconut milk and other organic substances are required for maximum growth. With cell suspensions the effect of deproteinized coconut milk is the same as that of raw coconut milk but, with callus cultures, deproteinized coconut milk gives greater yield. There are no obvious differences in yield of callus derived from root, hypocotyl, or cotyledon. Few differences in yield were obtained between cell suspension cultures from root, hypocotyl, and cotyledon but those from root gave the highest yield in dry weight. However, in the same medium, cells from the three origins are very similar in form and appearance. Some effects of different media on cell form and clumping are described. The yield in suspension culture is very high. Increase in cell number, fresh weight, and dry weight is about 100-fold in 12 days involving about six to seven divisions per cell.

The effect of 2,4-D and kinetin on the activity and isoenzyme pattern of various enzymes in cotyledon cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender)

1978 ◽  
Vol 56 (18) ◽  
pp. 2185-2195 ◽  
Author(s):  
Paul G. Arnison ◽  
W. G. Boll
Keyword(s):  
Acid Phosphatase ◽  
Phaseolus Vulgaris ◽  
Cell Suspension ◽  
Malate Dehydrogenase ◽  
Leucine Aminopeptidase ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Glutamate Oxaloacetate Transaminase ◽  
Cotyledon Cell ◽  
Isoenzyme Patterns

Changes in the activity and isoenzyme patterns of acid phosphatase, leucine aminopeptidase, glutamate-oxaloacetate transaminase, esterase, and malate and glutamate dehydrogenases were studied in cotyledon cell suspension cultures of Phaseolus vulgaris grown in the presence and absence of the growth regulators 2,4-dichlorophenoxyacetic acid and kinetin. With all enzymes studied, the pattern of isoenzymes and total enzymatic activity changed with the different phases of the culture cycle. In particular, the patterns of esterase, malate dehydrogenase, and glutamate dehydrogenase changed markedly with the inoculation of cells into fresh medium.The differences in isoenzyme patterns of cells grown with and without regulators were predominantly quantitative. However, certain minor isoenzymes of acid phosphatase, glutamate-oxaloacetate transaminase, esterase, and malate dehydrogenase were only detected in cultures grown in the presence of the regulators, while one isoenzyme of leucine aminopeptidase and two of esterase were unique to cells cultured in the absence of regulators.Three cathodic isoenzymes of acid phosphatase were released from wall material by 1 M NaCl. Such isoenzymes were also detected in the medium and in cytoplasmic extracts. Increase in the wall isoenzymes following inoculation into fresh medium was correlated with a decrease in anodic, cytoplasmic acid phosphatase.

The effect of 2,4-D and kinetin on peroxidase activity and isoenzyme pattern in cotyledon cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender)

1976 ◽  
Vol 54 (16) ◽  
pp. 1857-1867 ◽  
Author(s):  
Paul G. Arnison ◽  
W. G. Boll
Keyword(s):  
Phaseolus Vulgaris ◽  
Cell Suspension ◽  
Growth Regulators ◽  
Peroxidase Activity ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Peroxidase Isoenzymes ◽  
Cotyledon Cell ◽  
Cell Growth And Differentiation ◽  
Dichlorophenoxy Acetic Acid

Cotyledon cell suspension cultures of Phaseolus vulgaris were grown in the presence and absence of the growth regulators (2,4-dichlorophenoxy)acetic acid (2,4-D) and kinetin. Peroxidase (EC 1.11.1.7) activity was at a minimum during the phase of cell division and at a maximum during the phase of cell expansion. Both the pattern and activity of peroxidase isoenzymes changed during the culture cycle.Cells cultured without growth regulators showed increased peroxidase activity and changed isoenzyme patterns. Certain peroxidase isoenzymes were only present or prominent during specific phases of the culture cycle.The electrophoretic mobilities of peroxidase isoenzymes detected in the medium were not the same as those of the cytoplasmic isoenzymes. Cell cultures grown with and without growth regulators showed different patterns of medium peroxidase activity.Results are discussed in relation to the correlation of peroxidase activity with cell wall expansion and the possible role of peroxidase in cell growth and differentiation.

The effect of 2,4-D and kinetin on the morphology, growth, and cytochemistry of peroxidase of cotyledon cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender)

1976 ◽  
Vol 54 (16) ◽  
pp. 1847-1856 ◽  
Author(s):  
Paul G. Arnison ◽  
W. G. Boll
Keyword(s):  
Phaseolus Vulgaris ◽  
Cell Suspension ◽  
Peroxidase Activity ◽  
Morphological Characteristics ◽  
Enzyme Reaction ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Cell Age ◽  
Cotyledon Cell ◽  
Dichlorophenoxy Acetic Acid

Cotyledon cell suspension cultures of Phaseolus vulgaris were grown in the presence and absence of the growth regulators (2,4-dichlorophenoxy)acetic acid (2,4-D) and kinetin. Omission of the regulators changed the growth and cell division and produced striking changes in morphological characteristics. Cytochemical studies of peroxidase activity showed that the enzymes were mainly cytoplasmic in young cells and mainly associated with wall in older cells. No apparent differences in localization of activity were detected between the treatments, but cells cultured without the regulators showed a much more intense enzyme reaction. The location and increase of peroxidase, with increasing cell age and cell complexity, are consistent with the view that peroxidase activity is involved in cell wall expansion and differentiation.

Growth, and patterns of growth and division, in cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender)

1971 ◽  
Vol 49 (7) ◽  
pp. 1131-1139 ◽  
Author(s):  
Deng-Fong Liau ◽  
W. G. Boll
Keyword(s):  
Cell Suspension ◽  
Growth Regulation ◽  
Single Cells ◽  
Cell Suspension Cultures ◽  
Cell Types ◽  
Growth Cycle ◽  
Cell Number ◽  
Suspension Cultures ◽  
Growth Patterns ◽  
Bush Bean

Changes in cell number, cell size, fresh weight, and dry weight were recorded for the growth cycle of a serially subcultured cell suspension culture derived from hypocotyl of bush bean (var. Contender). The various cell types and growth patterns from single cells were also recorded. The culture showed unique features including a relatively sharp separation of a phase of exponential cell division from a phase of cell expansion. Complete separation of cell clumps gave a free cell culture, of viable cells, by day 16. The growth cycle, cell types, and growth patterns are compared with those in other plant cell suspension cultures. The potential value of the culture for study of growth regulation is discussed.

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