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1.
Jochen Berlin Sabine Sieg Dieter Strack Maria Bokern Hans Harms 《Plant Cell, Tissue and Organ Culture》1986,5(3):163-174
Red-violet cell suspension cultures of Chenopodium rubrum were found to accumulate the betacyanins amaranthin, celosianin and betanin and the betaxanthins vulgaxanthin I and vulgaxanthin II. Under a 16-h daylight regime the cells accumulated 0.3–0.4% betacyanins on a dry mass basis after 2–3 weeks of cultivation on the growth medium. Experiments to define a production medium for betacyanins failed with this habituated line. The accumulation could however be increased up to 1% or 100 mg betacyanins/1 by feeding tyrosine and by adaptation of the inoculum size to the nutrient concentration. 相似文献
2.
Resembling the lipids in the leaves and other green organs of intact plants, the lipids in photoautotrophic cell cultures of Chenopodium rubrum were found to contain high proportions of monogalactosyldiacylglycerols and digalactosyldiacylglycerols, as well as fair amounts of sulfoquinovosyldiacylglycerols and diacylglycerophosphoglycerols. Conversely, the heterotrophic cell cultures, from which the photoautotrophic cultures had been derived, contained only traces of these compounds. The heterotrophic cultures were rich in sterols, sterol esters, sterol glycosides, and esterified sterol glycosides. The lipids of photoautotrophic cell cultures contained higher proportions of constituent linolenic acid, but lower concentrations of linoleic acid than those of heterotrophic cultures. In the photoautotrophic cultures, as in green leaves, linolenic acid was predominantly estrified in monogalactosyldiacylglycerols and digalactosyldiacylglycerols. This investigation shows that it is possible to select strains of cell cultures, which are capable of grosing photoautotrophically, with the aim of activating the biosynthesis of specific metabolites. 相似文献
3.
On photoautotrophically grown, suspension-cultured cells of Chenopodium rubrum L. the electrical potential difference V
mand the electrical resistance across plasmalemma and tonoplast have been measured using one or two intracellular micro-electrodes. In a mineral test-medium of 5.8 mM ionic strength V
mvalues between 100 and 250 mV, 40% thereof between 170 and 200 mV, and a mean value (±S.E.M.) of 180.6±3.4 mV have been recorded. The average membrane input resistance R
mwas 269±36 M, corresponding to an average membrane resistivity r
mof 3.0 m2. V
mand r
mare sensitive to light, temperature, and addition of cyanide, suggesting the presence of an electrogenic hyperpolarizing ion pump, and are ascribed essentially to the plasmalemma. A hexose-specific saturable electrogenic membrane channel is identified through a decrease of V
mand r
mupon addition of hexoses. The hexoseconcentration-dependent depolarization V
msaturates at 92 mV and returns half-saturating concentrations (apparent k
mvalues) of 0.16 mM galactose, 0.28 mM glucose, and 0.48 mM fructose. The magnitude of V
mand r
mwell agrees with pertinent data from mesophyll cells in situ (where only V
mdata are available) and from photoautotrophic lower plant cells. However, V
mis markedly higher than reported for heterotrophically grown suspension cells of different higher plants (with which r
mdata have not been reported so far). It is concluded from the present study and a companion paper on water transport (Büchner et al., Planta, in press) that photoautotrophically grown Chenopodium suspension cells closely resemble mesophyll cells as to cell membrane transport properties.Abbreviations V
m
membrane potential(mV)
-
R
o
input resistance ()
-
R
m
membrane input resistance ()
-
r
m
specific resistance (resistivity) of the membrane (m2) 相似文献
4.
[3H]Dihydrozeatin supplied to photoautotrophically growing cell suspension cultures of Chenopodium rubrum was rapidly taken up and metabolized by the cells. The predominant metabolites in extracts of the cells were [3H]dihydrozeatin-O-glucoside and [3H]dihydrozeatin riboside-O-glucoside. Both these compounds could be shown to be compartmented within the vacuole, whereas [3H]dihydrozeatin and [3H]dihydrozeatin riboside, which were both present to a minor extent in cell extracts, were both present to a minor extent in cell extracts, were localized predominantly outside the vacuole. Analysis of the culture medium at the end of the 36-h incubation period showed that there had been an efflux of [3H]dihydrozeatin metabolites out of the cells. Whereas [3H]dihydrozeatin riboside was found to be the major extracellular [3H]dihydrozeatin metabolite, the O-glucosides of neither this compound nor [3H]dihydrozeatin could be detected in the medium. The differential compartmentation of [3H]dihydrozeatin metabolites found with the C. rubrum suspension-culture system is discussed with respect to possible mechanisms governing the metabolism of cytokinins in plants cells.Abbreviations (diH)Z
dihydrozeatin
- (diH) [9R]Z
9--D-ribofuranosyl dihydrozeatin
- HPLC
high-performance liquid chromatography
- ODS
octododecyl silica
- PEP
phosphoenolyruvate 相似文献
5.
Mixotrophic cell cultures of Chenopodium rubrum were found to synthesize 5 to 33 times more monogalactosyldiacylglycerols and 5 to 16 times more digalactosyldiacylglycerols than heterotrophic ones. The monogalactosyldiacylglycerols and digalactosyldiacylglycerols from mixotrophic cultures contained higher levels of linolenic acid as compared to heterotrophic cultures. It is concluded that the active synthesis of these galactolipids with high levels of constituent linolenic acid is associated with the onset of photosynthesis in plant cell cultures, as is the case in intact plants. 相似文献
6.
Chenopodium rubrum photoautotrophic cell suspensions were grown in plastic tissue culture dishes under photoautotrophic conditions. Growth was monitored by measuring cell number, packed cell volume, chlorophyll content and oxygen production. Such microtiter dishes are suitable systems for the serial assay of growth inhibition and various physiological effects (i.e. chlorophyll fluorescence, cell viability, oxygen production) of photoautotrophic cells as caused by herbicides and fungal phytotoxins. The applicability of the test system is discussed.Abbreviations pcv
packed cell volume
- fr.w.
fresh weight
- rpm
revol. per minute
- DMSO
dimethyl sulfoxide
- PMS
phenazine methosulfate
- NBT
nitro-blue tetrazolium chloride 相似文献
7.
Using the patch-clamp technique, we studied the action of charybdotoxin which blocks Ca(2+)-activated large-conductance K+ channels in animal tissue on the slow-activating (SV), Ca(2+)-activated cation channel in the vacuolar membrane of suspension-cells of Chenopodium rubrum L. The toxin reversibly reduced the vacuolar current with EC50 approximately 20 nM suggesting structural similarities between ion channels in animal and plant membranes. 相似文献
8.
The present experiments were carried out to investigate the effect of increased fluxes of H+ across the plasmalemma on glycolysis in heterotrophic cell suspension cultures of Chenopodium rubrum L. (1) Increased H+ influx was produced by adding glucose, 6-deoxyglucose, 2-deoxyglucose, or sodium fluoride. The net influx decreased to zero after 3 min. This recovery was accompanied by an increase in the rate of O2 uptake, but not of dark CO2 fixation. When glucose or fluoride were added, the increase of O2 uptake occurred without a decrease in the ATP/ADP ratio, and was large enough to provide the ATP that would be needed for compensatory H+ extrusion via the plasmalemma H+-ATPase. When 2-deoxyglucose was added, the rise of respiration was restricted by sequestration of phosphate and depletion of phosphorylated metabolites, the ATP/ADP ratio declined, and a slow net H+ influx started again after 4 min. (2) Alkalinisation of the medium to induce an H+ efflux resulted in rapid activation of dark CO2 fixation, but not of O2-uptake. (3) A stimulation of respiration or dark CO2 fixation was always accompanied by a decrease of phosphoenolpyruvate. This shows that the primary sites for regulation of glycolysis are pyruvate kinase and phosphoenolpyruvate carboxylase, respectively. (4) There was no consistent relation between glycolytic flux and triose-phosphates or hexose-phosphates. This shows that the reactions involved in carbohydrate mobilisation and the conversion of hexose-phosphates to triose-phosphates only have a secondary role in stimulation of glycolysis. (5) Phosphofructokinase will be stimulated as a consequence of the decrease in phosphoenolpyruvate. (6) The increase in glycolytic flux occurred independently of (in the case of 2-deoxyglucose and fluoride), or before (in the case of glucose), any increase of fructose-2,6-bisphosphate. When fructose-2,6-bisphosphate did increase (after supplying glucose), this was accompanied by an increase of triose-phosphate and fructose-1,6-bisphosphate, which otherwise remained very low. It is argued that fructose-2,6-bisphosphate increases as a consequence of the decrease of glycerate-3-phosphate, a known inhibitor of the synthesis of this regulator metabolite. However, activation of pyrophosphate fructose-6-phosphate phosphotransferase by fructose-2,6-bisphosphate does not play an obligatory role in the stimulation of glycolysis. 相似文献
9.
Expression of a sugar-transporter gene family in a photoautotrophic suspension culture of Chenopodium rubrum L. 总被引:1,自引:0,他引:1
Photoautotrophic suspension-culture cells of Chenopodium rubrum L. were shifted to mixotrophic growth by adding glucose to investigate whether the activities of plant sugar transporters, as well as the expression of the corresponding genes, are regulated in response to sugars. The rate of d-glucose uptake was shown not to be affected by mixotrophic growth in the presence of d-glucose. The polymerase chain reaction (PCR) technique was applied to amplify cDNA and genomic fragments from monosaccharide-carrier genes. Seven members of a monosaccharide-carrier family were identified of which three were found to be expressed in the suspension-culture cells. The expression of the monosaccharide-carrier genes was independent of the presence of d-glucose.Abbreviation PCR
polymerase chain reaction
We would like to thank Michaela Bittner, Rainer Ehneß and Monika Kammerer for skillful technical assistance and S. Buchhauser and H. Hallmer for photographic work. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 43) and by Fonds der Chemischen Industrie. 相似文献
10.
Lowering of the osmotic value of the medium has been reported previously to induce an oxidative burst in plant cells. This has been explained by a sequence of events, including solute influx, cellular swelling and the activation of stretch-regulated channels, triggering the production of reactive oxygen species. Moreover, it is known that the plant growth hormone auxin induces protoplast swelling. Together, these findings prompted the hypothesis that plant cells can respond to auxin-treatment with an oxidative burst. We tested this hypothesis using suspension cultured cells from Chenopodium rubrum L. and 2,7-dichlorodihydrofluorescein as the indicator for reactive oxygen species. An auxin-induced oxidative burst was found similar to an osmotically induced burst. Osmotic treatment consisted of a shift from 110 mOsm to 40 mOsm. The naturally occurring halogenated auxin 4-chloroindole-3-acetic acid was the most active compound tested, giving maximum rates of indicator oxidation corresponding to the formation of 4 x 10−1 5 mol H2 O2 cell−1 min−1 . Auxin analogous (10 μ M ) exhibited the following order of effectiveness: 4-chloride indoleacetic acid (100%), indoleacetic acid (80%), 2,4-dichlorophenoxyacetic acid (75%), 2-naphthylacetic acid (52%) and 1-naphthylacetic acid (47%). Benzoic acid (23%) was used as a control. Fusicoccin (35%) showed only slight stimulation in conjunction with complex kinetics. The detection of oxidative burst responses to 10 n M indoleacetic acid revealed a high sensitivity of the assay for auxin. Cell-free medium from aged batch cultures and light were also found to stimulate the production of reactive oxygen species. These data indicate that reactive oxygen species can transduce and integrate developmental and environmental signals and thus play a general role in plant growth regulation. 相似文献
11.
Water stress leads to a change of partitioning in favour of sucrose in heterotrophic cell suspension cultures of Chenopodium rubrum 总被引:1,自引:0,他引:1
Abstract. When heterotrophic cell suspension cultures of Chenopodium rubrum were subjected to water stress by incubation in increasing concentrations of sorbitol there was an immediate shrinking of the protoplast volume. Glucose uptake decreased in parallel. When the external water potential was greater than 0.5 MPa, a change in the partitioning of this incoming carbon was observed, resulting in a doubling of the ratio of sucrose to starch. This was accompanied by an increase in the concentration of the glycolytic intermediates and a particularly pronounced increase in the levels of fructose-2,6-bisphosphate and malate. The extractable activity of sucrose phosphate synthase was determined using a new method which overcomes the problem of uridine-diphosphate loss from the assay medium. It was found that sucrose phosphate synthase activity decreased in the water stressed cells. Therefore, the higher sucrose concentration in these cells is attributed to the increase in metabolite concentrations alone. 相似文献
12.
13.
Plant cell suspension cultures from Catharanthus roseus were investigated for their capability to dissimilate methionine or its analogs in order to reutilize the sulphane group for cysteine biosynthesis. Three steps have been described as prerequisites of this process: (a) oxidative degradation by the amino-acid oxidase of methionine giving rise to methanethiol production; (b) demethylation by methyltransferases leading to homocysteine and S-methylmethionine (c) replacement of the homocysteine sulphane sulphur by alkylthiol yielding methionine and free hydrogen sulphide. A reversal of the cystathionine pathway as a source of cysteine was ruled out because the cells lack cystathionine γ-lyase. The absence of this enzyme is compensated by the S-alkyl exchange of homocysteine with methylmercaptan. Hydrogen sulphide thus liberated is used for de novo synthesis of cysteine. The complete pathway can be catalyzed by the constitutive set of enzymes present in the higher plant. 相似文献
14.
Individual plants of Chenopodium rubrum were given differentnumbers of inductive cycles in a 12 h photoperiod and the patternof reproductive development was analysed after 40 d of growth.At least 2 inductive cycles are required to form any determinatereproductive organs and at least 12 cycles are required fornormal reproductive development. Individuals given a singleinductive cycle display a loss of apical dominance at thosenodes formed immediately after the treatment without the subsequentformation of any floral structures. Plants given between 2 and12 mductive cycles display both determinate reproductive organsand indeter minate vegetative shoots. The pattern of reproductivedevelopment on such plants depends upon the number of cyclesrelative to the developmental age of newly initiated primordia.It is suggested that the early events of floral induction mayinvolve a radical decrease in the ratio of auxin to cytokinin. 相似文献
15.
Summary Cell suspension cultures ofChenopodium rubrum have been grown for more than 2 years photoautotrophically with CO2 as sole carbon source. Average increase in fresh weight is appr. 600% within 14 days. The chlorophyll content of photoautotrophic cells (200 g/g fresh weight) is much higher than of photomixotrophic cells (50 g/g fresh weight). The photosynthetic activity of the cells (190 moles CO2×mg–1 chlorophyllXh–1) is comparable to the values found with intact leaves. As shown by short-term14CO2 photosynthesis, both, the photomixotrophic and the photoautotrophic cell suspension cultures assimilate CO2 predominantly via the Calvin pathway.Major differences were found with cells from either exponential or stationary phase of growth with regard to differential labelling of 3-phosphoglyceric acid, malate, sucrose and glucose/fructose.In vitro measurements of carboxylation reactions only partially corroborate our findings with14CO2 incorporation. The ratio of ribulosebisphosphate to phosphoenolpyruvate carboxylase activity is 4.7 for leaves of C.rubrum, 1.2 for photoautotrophic cells during stationary growth and 0.5 for cells during exponential growth phase, however, 0.18 was found for photomixotrophic cells. Though the14CO2 incorporation into 3-phosphoglyceric acid is clearly higher than into malate, thein vitro activity of phosphoenolpyruvatecarboxylase is 2–6 fold higher than that of ribulosebisphosphate carboxylase. We postulate that anaplerotic reactions of the tricarboxylic acid cycle are involved in the regulation of phosphoenolpyruvate carboxylase.Abbreviations 2,4-D
didilorophenoxyacetic acid
- EDTA
ethylene-diamine-tetraacetic acid
- fr. w.
fresh weight
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- PGA
3-phosphoglyceric acid
- PPO
2,5-diphenyloxazole
- PEP
phosphoenolpyruvate
- RuBP
nbulosebisphosphate 相似文献
16.
W. Hüsemann 《Plant cell reports》1983,2(2):59-62
The construction and operation of a continuous culture system for the propagation of cell suspensions from Chenopodium rubrum under photoautotrophic conditions has been described. A dilution rate of 0.16/day gave an equilibrium culture density of 1,100,000 cells/ml and a mean doubling time of 150 hours. During continuous culture steady state conditions with respect to nutrient uptake, cell protein and chlorophyll content, starch accumulation, in vitro activities of enzymes related to different metabolic pathways could be established. Photosynthetic CO2 assimilation of steady state cells was about 100 mol CO2/mg chlorophyll x hour. Dark CO2 fixation was 3% of the light values. 相似文献
17.
Jolanta Biesaga-Kocielniak Janusz Kocielniak Maria Filek Anna Janeczko 《Plant Cell, Tissue and Organ Culture》2008,94(2):139-147
The aim of this study was to produce suspension cultures of winter wheat directly from immature embryos bypassing the callus
stage, and to determine their capacity for growth and regeneration in comparison to suspension cultures produced from callus.
The study was carried out using Polish winter wheat varieties: ‘Grana’ and ‘Rosa’. Immature embryos were isolated, homogenized
and transferred directly to liquid medium supplemented with 2,4-D. Actively dividing cell cultures were obtained within 2 months
after the cultures were started. Suspension cultures from callus of immature embryos was also produced. With both cultivars,
faster growth was observed in the suspension cultures produced directly from embryos than in the suspensions produced from
callus. Metabolic activity was higher in the suspension culture produced directly from embryos than in the suspension derived
from callus only in ‘Grana’. The production of 1-amiocyclopropane-1-carboxylic acid (ACC), an ethylene precursor, was lower
in the suspension cultures produced directly from embryos than in the suspensions produced from callus. Morphogenic capacity
was significantly higher in aggregates derived directly from embryos than in aggregates derived from callus. With ‘Rosa’,
about one third of the aggregates derived directly from embryos regenerated shoots. Production of ACC was lower in ‘Rosa’
cell culture that regenerated then in other cell cultures that did not. Photosystem II reactions were more efficient in dark
green aggregates than in light green or pale green aggregates which were unable to regenerate. With the method presented,
wheat cell suspension cultures with a regeneration potential can be produced in 2 or 3 months less time than with traditional
methods. 相似文献
18.
Chalcone synthase (CHS) has been partially purified about 35-fold. Withdrawal of 2-mercaptoethanol after precipitation with ammonium sulfate led to higher stability during further purification steps. In order to determine CHS activity, two procedures [according to Schr?der et al. (1979) Plant Sci. Lett. 14, 281-286] were applied. The radioactivity extracted with ethyl acetate from the assay mixture (total products) was compared to 14C-labeled flavanone purified by TLC. The activity of CHS increased with bovine serum albumin (BSA) or 2-mercaptoethanol in the assay. Both effects were synergistic, but BSA did not promote "side products" as 2-mercaptoethanol did. BSA (10 mg ml-1) and 2-mercaptoethanol (1.4 mM) were components of the standard assay. Under these conditions, the CHS from Daucus carota had different pH optima for naringenin formation (7.9) and eriodictyol formation (6.8). The apparent Km values were 0.6 microM for 4-coumaroyl-CoA (pH 7.9), 7.7 microM for caffeoyl-CoA (pH 6.8), and 3.0 microM for malonyl-CoA (pH 7.9). Substrate inhibition was observed with 4-coumaroyl-CoA (greater than 10 microM) and malonyl-CoA (greater than 50 microM). The inhibitory activity of various flavonoids and related compounds (100 microM) was investigated. Naringenin and naringenin-chalcone inhibited eriodictyol formation totally and naringenin formation by 50%. In contrast, eriodictyol and eriodictyol-chalcone inhibited only eriodictyol formation by 40%. It was shown that the inhibition with naringenin was fully uncompetitive. These in vitro data support the view that the true substrate of CHS in D. carota is 4-coumaroyl-CoA. 相似文献
19.
Summary Synthesis of peanut agglutinin was induced in callus and cell suspension cultures of cotyledons of peanut (Arachis hypogaea L.). The lectin was synthesised in cultures through several passages. Biosynthesis of peanut agglutinin was regulated by the type and concentration of exogenous growth regulators and was positively correlated to the growth of the cultures, indicating that the agglutinin may have a role to play during cell growth. Movement of agglutinin from the cells into the medium not only facilitated easy isolation of the lectin but also provided a clue that it may probably serve as a defence molecule. The synthesized lectin purified from culture, was found to be biologically active, and was found to be comparable with the lectin from seeds, in terms of its electrophoretic mobility.Abbreviations BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- EDTA
ethylenediamine-tetraacetic acid
- HAU(s)
haemagglutination unit(s)
- IEF
isoelectric focusing
- KN
kinetin
- LS
Linsmaier and Skoog (1965) medium
- Mm
medium promoting minimum growth of cells
- MX
medium promoting maximum growth of cells
- NAA
naphthalene-1-acetic acid
- PBS
phosphate buffered saline
- PMSF
phenylmethylsulfonylfluoride
- PNA
peanut agglutinin
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- SHAA
specific haemagglutination activity
- TCA
trichloroacetic acid 相似文献
20.
W. Hüsemann 《Protoplasma》1982,113(3):214-220
Summary This communication describes the construction and operation of an airlift fermenter for the photoautotrophic growth of cell suspension cultures fromChenopodium rubrum. The basic batch culture unit provides a culture of 1.51 volume, sufficient to permit frequent aseptic sampling. It can be maintained at any desired temperature and aerated to different extents. Using an initial cell density of about 400,000 cells per ml suspension, the increase in cell number is 270% after a 14 days' growth period, although the stationary phase of growth is not yet reached. The transfer of photoautotrophic cell suspensions fromChenopodium rubrum from stationary growth into the large volume of fresh culture medium in the airlift fermenter results in an immediate protein formation, followed by an exponential phase of cell division, whereas rapid chlorophyll accumulation is delayed by 2 days.The growth capacities of photoautotrophic fermenter cultures including protein and chlorophyll formation as well asin vitro activities of the ribulosebisphosphate carboxylase and the phosphoenolpyruvate carboxylase are greatly lower as compared to photoautotrophic cells propagated in standard two-tier culture vessels using 30 ml culture medium. However the pattern of change in the activities of carboxylation enzymes is quite similar in both culture systems.Photoautotrophic cell suspensions fromChenopodium rubrum grown in an airlift fermenter assimilate about 90 mol CO2/mg chlorophyll × hour. Dark CO2 fixation is about 1.5% of the light values.Abbreviations
PEF
phosphoenolpyruvate
-
RuDP
ribulosebisphosPhate
-
NS
ground glass joints of standardized size made from Duran glass, Schott, Germany 相似文献