Growth and metabolism of cells and tissue of jack pine (Pinus banksiana). 3. Growth of cells in liquid suspension cultures in light and darkness

1976 ◽  
Vol 54 (5-6) ◽  
pp. 456-467 ◽  
Author(s):  
D. J. Durzan ◽  
V. Chalupa
Keyword(s):  
Pinus Banksiana ◽  
Basal Medium ◽  
Continuous Light ◽  
Cell Suspension Cultures ◽  
Jack Pine ◽  
Suspension Cultures ◽  
Growth Patterns ◽  
Final Size ◽  
Daughter Cells ◽  
Liquid Suspension

Friable, jack pine (Pinus banksiana Lamb.) callus on agar plates with a defined nutrient medium was transferred to a liquid medium of the same composition to establish cell suspension cultures. Dry weights and patterns of growth for daughter cells were estimated in continuous light or darkness, with or without conditioned media and arginine supplement. In all treatments, growth was near-exponential. Greatest final size was obtained with basal medium under continuous light.The distribution of clump sizes at the later stages of growth fits a stochastic model reflecting two types of daughter cell behavior described by the probability of one type remaining with the clump as opposed to sloughing off into the medium. In light, cells contained many more chloroplasts, thicker cell walls, and more compacted clumps than in darkness: yet cellular clumping patterns both in light and darkness were largely similar. Nearly 20% of the clumps showed polarity and symmetry and contained actively streaming suspensor-like cells. The remaining clumps were spherical and produced unorganized growth patterns. The progress of growth was eventually dominated by the tendency of daughter cells to remain attached to clumps compared with the initial tendency for clumps to fragment and cells to be released into the medium.

Growth and metabolism of cells and tissue of jack pine (Pinus banksiana). 6. Free nitrogenous compounds in cell suspension cultures of jack pine as affected by light and darkness

1976 ◽  
Vol 54 (5-6) ◽  
pp. 496-506 ◽  
Author(s):  
D. J. Durzan ◽  
V. Chalupa
Keyword(s):  
Free Amino ◽  
Pinus Banksiana ◽  
Daughter Cell ◽  
Growth Habit ◽  
External Medium ◽  
Cell Suspension Cultures ◽  
Jack Pine ◽  
Unit Weight ◽  
Nitrogenous Compounds ◽  
Final Weight

The increases of dry weights for cell suspensions of jack pine (Pinus banksiana Lamb.) callus grown in light or darkness were near-exponential, but the final weight was 10% less for those grown in darkness. Per unit weight, free amino acid N accumulated in cells in light but was lost from cells in darkness. Composition was dominated by high levels of glutamine, arginine, and alanine N. Several compounds, not usually present in callus on agar, or in the donor seedlings, were detected in cells.The composition of cells in relation to the external medium altered markedly. Individual nitrogenous compounds correlated with overall growth rates. Correlations between rates and composition reflected the distribution of clump sizes, i.e. differential daughter cell adhesion. Moreover, depending on the type of N compound studied, the ratio of endogenous to exogenous N either remained constant or varied, to reflect the growth habit of the suspension. Ratios for free glutamine N and monosubstituted guanidines were responsive to light and darkness and complemented one another.

Growth and metabolism of cells and tissue of jack pine (Pinus banksiana). 5. Changes in free arginine and Sakaguchi-reactive compounds during callus growth and in germinating seedlings of similar genetic origin

1976 ◽  
Vol 54 (5-6) ◽  
pp. 483-495 ◽  
Author(s):  
D. J. Durzan ◽  
V. Chalupa
Keyword(s):  
Growth Rates ◽  
Pinus Banksiana ◽  
Total Content ◽  
Dry Weight ◽  
Defined Medium ◽  
Jack Pine ◽  
Unit Weight ◽  
Final Size ◽  
Relative Growth Rates ◽  
Friable Callus

In friable callus, grown on a defined medium with arginine, and in jack pine seedlings, from which callus was derived, at least 18 Sakaguchi-reactive compounds and two N-phosphoryl derivatives were detected. When final size of callus was reached, the N in these substances contributed 0.1% to the total gain in dry weight and less than 0.6% to the gain in N. This was equivalent to a loss of 3 to 7% of the arginine N in the medium. For each callus, the total content of Sakaguchi-reactive compounds increased as the relative growth rates for weight and volume decreased. The increase was more pronounced when daughter cells adhered to clumps than at earlier stages when cells separated from one another. On a unit weight basis and as the cultures reached final size, the total Sakaguchi-reactive substances fell as γ-guanidinobutyric acid increasingly dominated the guanidine fraction. Exogenously supplied γ-guanidinobutyric acid caused a decrease in the final size of the cultured calluses when compared with calluses supplied with arginine. Levels of some of the remaining unidentified guanidines correlated positively with growth rates of calluses. By contrast, during the first 10 days of seedling growth Sakaguchi-reactive compounds contributed 12% of the net gain in N. The sequence of dominance of guanidines during growth was similar in both callus types and in seedlings, although some differences were noted.

Growth and metabolism of cells and tissue of jack pine (Pinus banksiana). 2. The quantitative analysis of the growth of callus from hypocotyls and radicles

1976 ◽  
Vol 54 (5-6) ◽  
pp. 446-455 ◽  
Author(s):  
V. Chalupa ◽  
D. J. Durzan ◽  
C. Vithayasai
Keyword(s):  
Growth Rates ◽  
Pinus Banksiana ◽  
Nitrogen Sources ◽  
Dry Weight ◽  
Growth Function ◽  
Jack Pine ◽  
Final Size ◽  
Relative Growth Rates ◽  
Green Callus ◽  
Food Reserves

Jack pine (Pinus banksiana Lamb.) calluses from hypocotyls and radicles of 5-day-old seedlings were established when the yield of callus from the donor was greatest compared with other stages of germination. Effects of macroelements, organic nitrogen sources, growth regulators, temperature, light, and carbon dioxide on the fresh and dry weights of callus indicated that the medium formulated was optimal. Although tissues assimilated exogenous sources of N, they also depended upon the food reserves that were mobilized during germination as the density (dry weight per cubic centimetre) of calluses gradually fell.Initially, the proliferation and release of cells from their neighbors was mainly on the outer surface of tissue. Eventually all cells appeared to divide to yield a friable, green callus with cell clumps that were easily dissociated from one another. Some clumps, especially those below the surface and near vascular tissues, were filamentous and exhibited polarity resembling the early stages of pine embryogeny.The growth form of hypocotyl callus was autocatalytic, whereas radicle callus was exponential for both dry weight and volume. Final size was greater in hypocotyl callus. In both systems, the relative growth rates of weight and volume correlated negatively but significantly to water intake. Average parameters of growth rates were calculated using Richards's comprehensive growth function.

Growth and metabolism of cells and tissue of jack pine (Pinus banksiana). 7. Observations on cytoplasmic streaming and effects of L-glutamine and its analogues on subcellular activities

1976 ◽  
Vol 54 (5-6) ◽  
pp. 507-517 ◽  
Author(s):  
D. J. Durzan ◽  
G. Bourgon
Keyword(s):  
Pinus Banksiana ◽  
Cytoplasmic Streaming ◽  
Basal Medium ◽  
Jack Pine ◽  
Concomitant Increase ◽  
Low Levels ◽  
Nuclear Behaviour ◽  
Transvacuolar Strands ◽  
Nucleolar Size ◽  
Streaming Motion

Cells from jack pine (Pinus banksiana Lamb.) callus and suspension cultures were observed by cinemicrography to evaluate cytoplasmic streaming, motion of organelles, and nuclear behaviour under glutamine stress. Streaming rates were greatest in the largest cells. Rates were reduced in smaller cells, where transvacuolar strands were not as prominent and displacements of organelles were not strongly affected by cyclosis. During increased rates of cytoplasmic streaming, chloroplasts tended to associate with each other and mitochondria often changed direction and shape.The addition of 400 ppm L-glutamine to a defined basal medium increased the linear velocity of organelles from 2-7 microns (μ)/s to 7–12 μ/s, whereas D-glutamine eventually stopped cytoplasmic streaming, with a concomitant increase in nucleolar size. While additions of low levels of α-N-acetyl-L-glutamine and isoglutamine stimulated streaming, this occurred usually without other significant morphological effects. Cytoplasmic movements that were induced by exposure of cells to L-glutamine were intimately involved with the fine strands and with nuclear and nucleolar behavior. Movements ceased when strands were disrupted under nutritional stress and by inhibitory responses to D-glutamine.

scholarly journals Effect of Sucrose and PEG Concentration on Embryo Maturation and Hyperhydration in Date Palm Suspension Cultures

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 462B-462 ◽  
Author(s):  
Shatha Matar Al-Matar ◽  
Mostafa M. Abo El-Nil ◽  
Jameel Al-Khairy ◽  
Gerald Klingaman
Keyword(s):  
Molecular Weight ◽  
Date Palm ◽  
Sucrose Concentration ◽  
Basal Medium ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Cultivar Differences ◽  
Ms Medium ◽  
Embryo Maturation ◽  
Two Factors

Cell suspension cultures of four date palm cultivars were established, namely, Niboat Saif, Madjhool, Sukarri, and Berhi. In this study, two factors were tested for their effect on embryo maturation and hyperhydration. The effect of sucrose concentration was assessed by inoculating 0.5 g of embryogenic callus into a liquid MS basal medium supplemented with 10 mg/L inositol, 3 mg/L glycine, 20 mg/L glutamine, and 0, 20, 30, 40, 50 g/L sucrose. Polyethylene glycol (PEG) concentration effect on embryo maturation and hyperhydration was tested. PEG (molecular weight 7000–9000) was added at concentrations of 0, 10, 30, and 60 g/L to the date palm suspension cultures. Cultures were examined and subcultured every 3 weeks for 2 months. Embryos formed were then transferred to a solid MS medium supplemented with 10 mg/L inositol, 3 mg/L glycine, 5 mg/L glutamine, and 30 g/L sucrose. The number of embryos germinated from each treatment was counted to compare cultivar differences. Preliminary data suggests that the medium containing 30 g/L sucrose is most effective for embryo maturation, and those embryos germinated when transferred to a solidified MS medium. The study found that incorporating PEG into the medium reduced the hyperhydration of date palm tissues. The various cultivars reacted differently to the treatments employed.

scholarly journals OPTIMAL CONDITIONS FOR CELL SUSPENSION CULTURES OF THLASPI CAERULESCENS AND BRASSICA NAPUS

HortScience ◽  
1995 ◽  
Vol 30 (2) ◽  
pp. 186b-186
Author(s):  
W. Bundithya ◽  
S.L. Kitto
Keyword(s):  
Basal Medium ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Thlaspi Caerulescens ◽  
Cell Suspensions ◽  
Ms Medium ◽  
Optimal Conditions ◽  
Inoculation Density ◽  
Gyratory Shaker ◽  
High Concentrations

Thlaspi caerulescens (Brassicaceae), known as a Zn hyper accumulator, is able to accumulate and tolerate Zn, Ni, Cu, and Cd at high concentrations in its biomass. We are examining the feasibility of using cell suspensions of T. caerulescens and B. napus to study the effect of selected heavy metals on growth and nutrient uptake. Callus was initiated by culturing seedlings on basal medium containing MS salts supplemented with MS or B5 vitamins, 1, 2, 5, or 10 mg 2,4-D/liter, and 0.7% Phytagar. Cell suspensions were initiated by transferring calli to liquid basal medium containing MS or B5 vitamins, and 1 or 2 mg 2,4-D/liter, and were incubated on a gyratory shaker at 120 rpm. Growth of suspensions inoculated at 0.2, 0.4, or 0.6 g/25 ml was monitored for 13 days. Optimal conditions required to initiate and maintain suspension cultures of T. caerulescens and B. napus include MS medium supplemented with B5 vitamins and 1 mg 2,4-D/liter, an inoculation density of 0.4 g/25 ml, and a 2-week subculture schedule.

scholarly journals Influence of an Indirect Precursor on Podophyllotoxin Accumulation in Cell Suspension Cultures of Podophyllum hexandrum

2013 ◽  
Vol 22 (2) ◽  
pp. 171-177 ◽  
Author(s):  
Anrini Majumder
Keyword(s):  
Cell Suspension ◽  
Biomass Yield ◽  
Basal Medium ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Podophyllum Hexandrum ◽  
Naturally Occurring ◽  
Lignan Biosynthesis ◽  
Half Strength ◽  
Commercial Source

Podophyllum hexandrum is an endangered, commercial source of a naturally occurring lignan podophyllotoxin, which is a precursor of potent anticancer drugs. Cell suspension cultures were established using four-year-old cell line RC4 of P. hexandrum in liquid P1 medium [half strength B5 basal medium supplemented with 1% (w/v) sucrose, 2,4-D (1 mg/l), GA3 (1 mg/l) and BA (0.1 mg/l)]. The effect of different concentrations (100, 250 and 500 mg/l) of tryptophan, an indirect precursor of lignan biosynthesis, was studied on cell suspension cultures. Cultures accumulated 2.7 times more podophyllotoxin than untreated control cultures when treated with 250 mg/l tryptophan. Biomass yield was, however not improved following the addition of tryptophan. DOI: http://dx.doi.org/10.3329/ptcb.v22i2.14207 Plant Tissue Cult. & Biotech. 22(2): 171-177, 2012 (December)

Growth and metabolism of cells and tissue of jack pine (Pinus banksiana). 4. Changes in amino acids in callus and in seedlings of similar genetic origin

1976 ◽  
Vol 54 (5-6) ◽  
pp. 468-482 ◽  
Author(s):  
D. J. Durzan ◽  
V. Chalupa
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Free Amino Acid ◽  
Growth Rates ◽  
Free Amino ◽  
Pinus Banksiana ◽  
Dry Weight ◽  
Final Size ◽  
Plant Parts ◽  
Daughter Cells

Growth rates of callus from hypocotyls and radicles of jack pine (Pinus banksiana Lamb.) seedlings were associated with the differential release and adherence of daughter cells. From 18 to 40% of the N in the medium was assimilated through the increasing surface area of callus and contributed nearly 6% of the total gain in dry weight. Sequential changes in free amino acid N leading to final size were similar for callus from both plant parts. Changes in N correlated significantly with growth rates and callus density (dry weight per cubic centimetre). When free amino acid N was expressed as a percentage of the total soluble N, correlations signified the relative proportions of the amino acid pool that gave different growth-rate forms. The balance of N metabolism during the autocatalytic growth of hypocotyl callus was under the influence of free glutamine, whereas the exponential growth of radicle callus was dominated by the synthesis of asparagine N. In callus from both plant parts 80 to 87% of the net gain in N was recovered in the bound (protein and nucleic acid) fraction. Percentage of free glutamine N decreased with the increase in bound N.Ways in which free amino acid N interacted with growth rates indicated that final size may be usefully defined not merely as an extreme of a growth function but in more sequential kinetic terms supported by a series of correlation coefficients. Correlations revealed the specific points at which free amino acids interacted strongly with rates.When amino acid N in callus was compared with N in seedlings from which callus was derived, the trends were strikingly similar. In seedlings the succession of amino acids was linked more to the uptake of N from the gametophyte by cotyledons and daughter cells in growing tissues rather than to their morphogenesis. Differences related mainly to the greater ability of the seedling to synthesize protein and to maintain higher levels of total soluble N than in callus.

scholarly journals Optimization of callus and cell suspension cultures of Barringtonia racemosa (Lecythidaceae family) for lycopene production

2011 ◽  
Vol 68 (1) ◽  
pp. 69-76 ◽  
Author(s):  
Mandana Behbahani ◽  
Mehrnaz Shanehsazzadeh ◽  
Mohamad Javad Hessami
Keyword(s):  
Cell Suspension ◽  
Callus Induction ◽  
Leaf Explants ◽  
Basal Medium ◽  
Cell Suspension Cultures ◽  
Reversed Phase ◽  
Suspension Cultures ◽  
Cell Culture Process ◽  
Lycopene Production ◽  
Lycopene Content

Lycopene is present in a range of fresh fruits and vegetables, especially in the leaves of Barringtonia racemosa. The traditional lycopene extraction from the plant is being employed instead of an easy propagation technique like cell culture process from the leaf explants. We intend to assess how lycopene could be extracted via tissue culture under light (illuminance: 8,200 lux under white fluorescent lamps, photoperiod 16 h per day at 25ºC) and dark. Leaf explants of Barringtonia racemosa were cultured on modified Murashige and Skoog (MS), Woody Plant Medium (WPM) and B5 media, supplemented with different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D). Optimal conditions for callus induction and maintenance under both dark and light were investigated, and growth and lycopene accumulation were evaluated. Among media with different concentrations of 2,4-D, fast growing, friable callus initiated within three weeks after culturing on WPM basal medium supplemented with 2.0 mg L-1 (weight per volume) of 2,4-D, whereas callus induction in explants cultured on all other media started only after five weeks. Calli were subcultured once every fortnight. Pale yellow and green calli developed under conditions of dark and light respectively were then selected for evaluation of their lycopene contents. An improved reversed phase of high performance liquid chromatography (HPLC) method was used for a selective chemical determination of the lycopene content. Light induced lycopene production; and likewise maximum lycopene level incubated in light was higher than those incubated in darkness. The best growth rates of callus and cell suspension were achieved in WPM and B5 media respectively. The production of lycopene was growth-dependent through analysis of growth and lycopene content of both callus and cell suspension cultures.

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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