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1.
The levels of purine and pyrimidine nucleotides in suspensioncultures of Catharanthus roseus were determined 24 h after stationary-phasecells were transferred to fresh complete (‘+Pi’)or phosphate-deficient (‘–Pi’) Murashige-Skoogmedium. The levels of ATP, GTP, UTP and CTP were from approx.3 to 5-fold greater in the cells grown in ‘+Pi’medium than in the cells grown in ‘–Pi’ medium.The levels of almost all other nucleotides were slightly higherin the cells in ‘+Pi’ medium. The rates of de novoand salvage biosynthesis of purine and pyrimidine nucleotideswere estimated from the rates of incorporation of radioactivityfrom [14C]formate, [2–14C]glycine, NaH14CO3, [6–14C]orotate,[8–14C]adenine, [8–14C]adenosine, [2–14C]uraciland [2–14C]uridine. The results indicated that the activityof both the de novo and the salvage pathway was higher in thecells in ‘+Pi’ medium than in the cells in ‘–Pi’medium. The rate of degradation estimated from the rate of releaseof 14CO2 from labelled purines and pyrimidines indicated thatdegradation of uridine was significantly reduced in the cellsin ‘+Pi’ medium, but no significant difference wasfound in the degradation of adenine, adenosine and uracil. Thepossible role of Pi in the control of the biosynthesis of nucleotidesand in the degradation of uridine is discussed. Catharanthus roseus, Madagascar periwinkle, suspension culture, inorganic phosphate, nucleotides, purines, pyrimidines, biosynthesis, degradation  相似文献   

2.
The levels of free amino acids in suspension cultures of Catharanthusroseus were monitored for 96 h after stationary-phase cellswere transferred to fresh complete (‘+Pi’)  相似文献   

3.
The uptake and metabolism of sugars by suspension-cultured Catharanthusroseus cells were investigated. Substantially all the sucrosein the culture medium was hydrolyzed to glucose and fructosebefore being taken up by the cells. The activity of invertasebound to cell walls, determined in situ, was high at the earlystage of culture. Glucose was more easily taken up by the cellsthan was fructose. Tracer experiments using [U-14C]glucose and[U-14C]fructose indicated that glucose is a better precursorfor respiration than fructose, while fructose is preferentiallyutilized for the synthesis of sucrose, especially in the earlyphase of cell growth. Possible metabolic routes of sugar insuspension-cultured Catharanthus roseus cells are discussedin the context of these results. Catharanthus roseus, Madagascar periwinkle, suspension culture, sucrose, glucose, fructose, metabolism, glycolysis  相似文献   

4.
The metabolism of [2-14C]thymine, [2-14C]thymidine, [2-14C]uraciland [14C]uridine was investigated in protoplasts obtained fromsuspension cultures of Catharanthus roseus. Most of the exogenouslysupplied thymine, thymidine and uracil was degraded, and salvageof these pyrimidines accounted for 5–36 per cent of thetotal amount of 14C-labelled precursors which was metabolized.However, more than 80 per cent of the labelled uridine was utilizedfor the biosynthesis of nucleotides and nucleic acids, and therest was degraded. In contrast to the results from protoplastsof sugar cane cells in suspension culture, the data indicatethat protoplasts possess a pathway for the degradation of pyrimidines,and that the overall metabolism of these pyrimidines in protoplastsis very similar to the metabolism in the intact cells. Catharanthus roseus, madagascar periwinkle, protoplasts, pyrimidine metabolism  相似文献   

5.
A microsomal flavonoid 3'-hydroxylase (F3'H) catalyzing themetabolism of naringenin to eriodictyol in Citrus sinensis (L.)Osbeck cv. ‘Hamlin’ cell suspension cultures wasshown to be a cytochrome P450 enzyme. This reaction requiredO2 and NADPH and was inhibited by CO, with partial reversalof CO-inhibition by light at 450 nm. Cytochrome P450 contentranged from 10–20 pmol (mg microsomal protein)–1.The F3'H reaction was shown to be linear in regard to proteinconcentration between 2.5 and 25 µg of microsomal protein.The optimum pH for the reaction was 7.4–7.6 and the temperatureoptimum was between 30 and 37°C. The apparent Km and Vmaxfor naringenin were 24 µM±3.2 and 81.4±7.9pmol eriodictyol min–1 (mg protein)–1, respectively.The microsomal F3'H was also capable of forming dihydroquercetinfrom dihydrokaempferol (40 pmol min–1 (mg protein)–1)and of quercetin from kaempferol (3.25 pmol min–1 (mgprotein–1). Cytochrome c and ketoconazole were the bestinhibitors of WH activity followed by piperonyl butoxide anda-naphthoflavone. Light was shown to be an inducer of the F3'Halmost doubling the specific activity and increasing the microsomalcytochrome P450 content by 30% over that of dark grown cells.F3'H activity was also confirmed in microsomal preparationsof young (new flush) leaves from ‘Hamlin’ treesand flavedo of ‘Hamlin’ oranges, ‘Marsh’grapefruit, and ‘Lisbon’ lemon. No activity wasobserved in older, hardened leaves and albedo of all the fruittested. Initiation of embryogenesis in the ‘Hamlin’cell suspension cultures by switching from a sucrose mediumto a glycerol-based medium resulted in the down-regulation ofF3'H. 1Mention of a trademark, warranty, proprietary product, or vendordoes not constitute a guarantee by the U.S. Department of Agricultureand does not imply its approval to the exclusion of other productsor vendors that may also be suitable.  相似文献   

6.
The effects of abscisic acid (ABA) treatments on the surfaceelectrical properties of cells and isolated protoplasts fromCatharanthus roseus cell suspension cultures were studied byelectrophoretic mobility and 9-aminoacridine (9AA) fluorescencemeasurements. The surface charge densities of the cells andprotoplasts estimated from electrokinetic data were –0.064Cm–2and –0.048 C m–2 respectively. These values wereclose to that estimated by 9AA fluorescence technique i.e.,–0.053 Cm–2 for the cells and –0.041 Cm–2for the isolated protoplasts accordingly. The net negative surfacecharge density decreased after application of 10 µM and50 µM ABA in both cells and protoplats, the more pronouncedeffect being observed at 10 µM ABA. When 100 µMABA was supplemented to the cell suspension culture the oppositeeffect was observed. The average charge density increased to–0.074 C m–2 for the cells, and to –0.055C m–2 for protoplasts, as revealed from the 9AA measurements.The results are discussed in terms of specific concentrationdependent ABA-induced alterations of the electrostatic propertiesof cell and protoplast membranes. (Received December 12, 1994; Accepted April 3, 1995)  相似文献   

7.
In order to clarify the metabolic adaptation of respiratorypathways in plants to limited levels of Pi, the effects of long-termstarvation of Pi on the activities of various enzymes relatedto respiratory metabolism were examined in suspension-culturedCatharanthus roseus cells. When the activities were expressedas units per g fresh weight, only those of phosphoenolpyruvate-hydrolyzing(PEP-hydrolyzing) enzyme (which may possibly be equivalent tothe acid phosphatase activity derived from vacuoles) and PEPcarboxylase were higher in the Pi-starved cells than in controlcells. Activities of other enzymes in the Pi-starved cells werelower than or similar to those of the control cells. Time-coursestudies indicated that PEP-hydrolyzing activity was inducibleby starvation of Pi. However, in contrast to the results reportedby Duff et al. [(1989a) Plant Physiol. 90: 1275.], fluctuationsin the activity of PP1:fructose-6-phosphate 1-phosphotransferaseduring starvation of Pi were similar to those in levels of phosphofructokinaseand 6-phosphogluconate dehydrogenase. These data suggest thatthe concept of the phosphate starvation-inducible ‘bypasses’,which are engineered via the coarse control (i.e., induction)of specified enzymes and were proposed initially by Duff etal. in Brassica nigra cells, is not directly applicable to Catharanthusroseus cells in suspension. Tracer experiments using [U-14C]glutamineindicated that a significant proportion of respiratory substratescould be supplied from the enlarged pool of amino acids duringstarvation of Pi. These assumptions are supported by the observedfluctuations in levels of free amino acids and of protein inP1-fed and P1-deficient Catharanthus roseus cells. 1Part 41 in the series ‘Metabolic Regulation in PlantCell Cultrue’ 2Present Address: Morinaga Mild Industry, 5-1-83, Higashihara,Zamma-shi, Kanagawa, 228 Japan  相似文献   

8.
The effects of sugars on the glucosylation of exogenous hydroquinone(HQ) was investigated by supplying individual sugars simultaneouslywith HQ to a suspension culture of Catharanthus roseus cells.The production of arbutin was enhanced as much as 2- to 3-foldby sucrose or glucose at concentrations of up to 6%, with theenhancement being directly dependent on the concentration ofthe sugar. The exogenously added sugar was not metabolized andremained unchanged. Sorbitol also promoted the production ofarbutin in a similar manner. Sucrose improved the viability of cells and, in cultures suppliedwith sucrose and HQ, the activity of UDP-glucose: hydroquinoneglucosyltransferase increased over a much longer period of timethan that in control cultures supplemented with HQ only. (Received December 11, 1989; Accepted March 26, 1990)  相似文献   

9.
Marked changes in the activity of the ‘de novo’and ‘salvage’ pathways of pyrimidine biosynthesisduring growth of Vinca rosea cells in a batch suspension culturewere observed. The activity of these pathways was investigated by determiningthe contribution of 14C of [2-14Cluracil, 12-14Cluridine. and[6-14Clorotate to the cell constituents and by measuring theactivity of the several enzymes of these pathways. During the lag phase of the culture, ‘uracil-’ and‘uridine-salvage’ pathways made the predominantcontribution to nucleotide biosynthesis, but, following theinitiation of cell division, the ‘de novo’ pathwayfor nucleotide biosynthesis operated appreciably. These results suggest that nucleotide synthesis during cellgrowth in a suspension culture can be divided into two stages:a ‘turnover stage’, during the lag phase of cellgrowth, and a ‘true biosynthetic stage’, which isinitiated in the cell division phase.  相似文献   

10.
Barley (Hordeum vulgare L., cvs Golf, Mette, and Laevigatum)was grown under nitrogen limitation in solution culture untilnear maturity. Three different nitrogen addition regimes wereused: in the ‘HN’ culture the relative rate of nitrate-Naddition (RA) was 0·08 d–1 until day 48 and thendecreased stepwise to, finally, 0·005 d–1 duringgrain-filling; the ‘LN’ culture received 45% ofthe nitrogen added in HN; the ‘CN’ culture was maintainedat RA 0·0375 d–1 throughout. Kinetics of net nitrateuptake were measured during ontogeny at 30 to 150 mmol m–3external nitrate. Vmax (which is argued to reflect the maximuminflux rate in these plants) declined with age in both HN andLN cultures. A pronounced transient drop was observed just beforeanthesis, which correlated in time with a peak in root nitrateconcentration. Similar, but less pronounced, trends were observedin CN. The relative Vmax (unit nitrogen taken up per unit nitrogenin plants and day) in all three cultures declined from 1·3–2·3d–1 during vegetative growth to 0·1–0·7d–1 during generative growth. These values are in HN andLN cultures 15- to more than 100-fold in excess of the demandset by growth rates throughout ontogeny. Predicted balancingnitrate concentrations (defined as the nitrate concentrationrequired to support the observed rate of growth) were below6·0 mmol m–3 in HN and LN cultures before anthesisand then decreased during ontogeny. In CN cultures the balancingnitrate concentration increased during grain-filling. Apartfrom the transient decline during anthesis, most of the effectof ageing on relative Vmax can be explained in terms of reducedcontribution of roots to total biomass (R:T). The loss in uptakeper unit root weight is largely compensated for by the declinewith time in average tissue nitrogen concentrations. The quantitativerelationships between relative Vmax and R:T in ageing plantsare similar to those observed for vegetative plants culturedat different RAs. The data support the contention that the capacity for nitrateacquisition in N-limited plants is under general growth control,rather than controlled by specific regulation of the biochemicalpathway of nitrate assimilation. Key words: Barley, nitrogen concentration, root: total plant biomass ratio, Vmax  相似文献   

11.
Smith, J. I, Smart, N. J., Kurz, W. G. W. and Misawa, M. 1987.The use of organic and inorganic compounds to increase the accumulationof indole alkaloids in Catharanthus roseus (L.) G. Don cellsuspension cultures.—J. exp. Bot. 38: 1501–1506. The addition of sodium chloride, potassium chloride or sorbitolto 5–d–old cell suspension cultures of Catharanthusroseus stimulated an increase in the intracellular accumulationof catharanthine and other indole alkaloids within 48–72h. The magnitude of the response depended upon the concentrationof the compound added. The use of such inexpensive and readilyavailable compounds to increase the yields and reduce the requiredculture times has considerable potential for the productionof useful secondary metabolites from cell cultures of C. roseusand other plant species.  相似文献   

12.
The effects of exposure of up to 2 h with sulphur dioxide ona range of plant species was observed by measuring changes inthe rate of net photosynthesis under closely controlled environmentalconditions. Ryegrass, Lolium perenne ‘S23’ was thespecies most sensitive to SO2; significant inhibition was detectedat 200 nl l–1. Fumigations at 300 nl l–1 also inhibitedphotosynthesis in field bean (Vicia faba cv. ‘Three FoldWhite’ and ‘Blaze’) and in barley (Hordeumvulgare cv. ‘Sonja’). No effect was detected inwheat (Triticum aestivum cv. ‘Virtue’) at concentrationsup to 600 nl l–1 SO2, or in oil-seed rape (Brassica napuscv. ‘Rafal’) except at 800 nl l–1 SO2). Recoverycommenced immediately after the fumigation was terminated andwas complete within 2 h when inhibition had not exceeded 20%during the SO2 treatment. Key words: Sulphur dioxide, short-term fumigation, photosynthesis  相似文献   

13.
Incorporation of polyamine-spermidine into the nutrient mediumat 10–6 and 10–5 M concentrations stimulates pollen-tubegrowth in vitro in Catharanthus roseus L. G. Don. MGBG, an inhibitorof spermidine biosynthesis, at 0.5 x 10–3 and 1 x 10–3M concentrations reduced the percentage of germination as wellas tube growth and at a concentration of 1.5 x 10–3 Mgermination was totally inhibited. Pollen grains incubated inthe medium containing 1.5 x 10–3 M MGBG, when transferredto a fresh medium with 10–5 M spermidine, resulted in80% germination recovery, along with considerable tube growth.Experiments with actinomycin-D indicate that stimulation ofpollen-tube growth by spermidine may involve de novo synthesisof protein. Catharanthus roseus, pollen germination, tube growth, spermidine, MGBG, inhibition, actinomycin-D  相似文献   

14.
An investigation was carried out to examine the effect of aerationon the growth of Catharanthus roseus suspension cultures inairlift bioreactors. A high aeration rate (0·86 v.v.m.)was found to inhibit the growth of cultures. Venting culturesat a high rate with low oxygen content gas mixtures was equallyinhibitory to culture growth, showing that high aeration wasnot inhibitory as a result of oxygen toxicity. The dissolvedcarbon dioxide tension was found to be lower in cultures operatedat high aeration than those operated at low aeration. Supplyingexogenous CO2 to cultures at high aeration restored the CO2tension to values normally encountered at a low aeration rate,and was found to alleviate the inhibitory effects at high aeration.However, further increasing the CO2 supply to cultures was foundto be severely inhibitory to growth. Therefore, the growth ofC. roseus cultures is very sensitive to dissolved CO2 concentration,growth being inhibited at values either higher or lower thanan optimum. Key words: Aeration, carbon dioxide, Catharanthus roseus suspension culture  相似文献   

15.
Reducing the concentration of sucrose in the culture mediumover successive subcultures has been tested as a method forincreasing the ability of rose shoots grown in vitro (Rosa cvsIceberg and Peace) to take up CO2. Shoots maintained on ‘constant’10, 20 and 40 g I–1 sucrose showed decreased levels ofCO2 uptake at higher sucrose concentrations, although cv. Peacegrew least at 10 g l–1 and showed correspondingly lowamounts of CO2 uptake compared with 20 and 40 g l–1. Bothcultivars died when sucrose was omitted from the medium. Assucrose concentration was reduced in the medium, so CO2 uptakeof shoots initially cultured on 20 and 40 g l–1 sucrosewas found to increase, although a concentration of 10 gl –1sucrose seemed to be limiting, below which the growth and chlorophylllevels of shoots declined. Rosa hybrid, rose, shoot culture in vitro, photosynthetic ability, sucrose, infra-red gas analysis  相似文献   

16.
After removal of the embryo from developing seeds of Pisum sativum,the ‘empty’ ovules (seed coats without enclosedembryo) were filled with a solution (pH 5.5) containing mannitol(usually 400 mM) to which various salts were added. A solutioncontaining two isotopes ((a) [2H]-sucrose/[–14C]aminoisobutyricacid (AIB) or (b) [3H]valine/[14C]asparagine mixture) was administeredto the plant via the petiole subtending the fruiting node, and[2H]solute and [14C]solute unloading from the seed coat wasmeasured, in pulse-labelling experiments of about 5 h. The presenceof 25 or 50 mM K+ in the ‘empty’ ovule enhancedthe release of sucrose from the seed coat particularly duringthe first hours of the experiment, but the stimulating effectof K+ on the release of labelled solutes derived from aminoacids was much smaller. The presence of 25 mM CaCl2 did notaffect the release of sucrose or amino acids from the seed coat.The effect of K+ on sucrose and amino acid release is explainedas an inhibition of sucrose and amino acid resorption from theseed coat apoplast into seed coat cells, after unloading fromthe seed coat unloading sites. It is suggested that amino acidrelease is much less affected by K+ than sucrose release, becausefar less resorption of amino acids by seed coat parenchyma cellstakes place during amino acid transport into the seed coat cavity. Pisum sativum, pea, assimilate transport, assimilate unloading, seed-coat exudate, seed development, sucrose resorption, surgical treatment  相似文献   

17.
Nitrate provision has been found to regulate the capacity forChara corallina cells to take up nitrate. When nitrate was suppliedto N sufficient cells maximum nitrate uptake was reached after8 h. Prolonged treatment of the cells in the absence of N alsoresulted in the apparent ability of these cells to take up nitrate.Chlorate was found to substitute partially for nitrate in the‘induction’ step. The effects on nitrate reductionwere separated from those on nitrate uptake by experiments usingtungstate. Tungstate pretreatment had no effect on NO3uptake ‘induced’ by N starvation, but inhibitedNO3 uptake associated with NO3 pretreatment. Chloridepretreatment similarly had no effect on NO3 uptake ‘induced’by N deprivation, but inhibited NO3 uptake followingNO3 pretreatment. The data suggest that there are atleast two mechanisms responsible for the ‘induction’of nitrate uptake by Chara cells, one associated with NO3reduction and ‘induced’ by CIO3 or NO3and one associated with N deprivation. Key words: Nitrate, Chlorate, Chara corallina, Induction  相似文献   

18.
Photosynthetic development of Spinacia oleracea L. (spinach)cells was ‘triggered’ by halving the supply of sugarto a continuous culture. This was demonstrated between two steadystates at constant specific growth rate (4.2 ± 0.17 x10–3 h–1). As a direct consequence of this ‘trigger’the cells were in conditions of sugar famine, demonstrated byorganic carbon balance. Comparison of the biomass was made betweenthe two steady states. It was found that in sugar famine thebiomass contained 8 times as much chlorophyll and the ratioof photosynthesis to respiration in saturating conditions hadincreased 22 times, when compared to the biomass in conditionof sugar excess. All substrate carbon and organic factors were then removed fromthe fresh medium, carbon dioxide and photosynthetically activeradiation (PAR) were increased. In these photoautotrophic conditionsthree further steady states were established. The effects ofincreasing the dilution rate and then increasing PAR were examined.Growth was found to be limited by PAR. The photosynthetic yield Y of photoheterotrophic spinach cellsin fructose famine was estimated as 0.004 g kJ–1 PAR.In photoautotrophic conditions the maximum Y was 0.0018 g kJ–1.It was suggested that this lower yield was due to chloroplastdevelopment being arrested by an unknown factor.  相似文献   

19.
Techniques are described for the culture of developing barley(Hordeum vulgare L.) caryopses. Over a 7 d period in culturethe dry weights and the amounts of starch and protein increasedby at least twofold. Growth was sustained for at least 20 d.The effects of glutamine and cysteine on the amount and compositionof the hordein storage proteins were also studied. Glutaminestimulated total hordein accumulation but caused a disproportionateincrease in the amount of the S-poor ‘C’ hordeinswhen supplied at 100 mol m–3. Addition of cysteine at1·0 mol m–3 did not increase the amount of Srich‘B’ hordeins. The results suggest that althoughisolated caryopses are able to take up sucrose and glutamineand convert them to starch and protein there is some limitationin their ability to convert externally supplied cysteine intoproteins. Key words: Hordeum vulgare L., Caryopses, Glutamine, Cysteine, Storage proteins  相似文献   

20.
ERRATA     
Effects of coupled solute and water flow in plant roots withspecial reference to Brouwer's experiment. Edwin L. Fiscus. p. 71 Abstract: Line 3 delete ‘interval’ insert‘internal’. p. 73 Materials and Methods: line 6: delete ‘diversion’ insert ‘division’ line 9 equation should read Jv=Lp PRT(C0C1). 74 Last line of figure legend: 10–1 should read 10–11. 75 Line 11: delete ‘seems’ insert ‘seem’. le 1 column heading—106 should read 1011. 77 delete ‘...membrane in series of...’ insert ‘membranein series or...’ Delete final paragraph.  相似文献   

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