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1.
A line of S-(2-aminoethyl)-L-cysteine-resistant adenine-auxotrophiccells (AdAECr strain) was isolated from adenine-auxotrophiccells (Ad strain) of Datura innoxia Mill by a stepwiseselection method. AdAECr and Bl cells, which were clonedfrom the original AdAECr cells, were able to grow activelyon medium that contained 10 mM S-(2-aminoethyl)-L-cysteine (AEC),whereas the growth of Ad cells ceased completely in thepresence of 0.5 mM AEC. The resistant phenotype has been maintainedfor at least 10 months in culture on medium without AEC. Levels of free lysine in AdAECr and Bl cells were similarto that in Ad cells. By contrast, the level of free AECin AdAEC cells was 10-fold lower than in Ad cellsand no free AEC was detectable in Bl cells. However, acid hydrolysisof extracts from AdAECr and Bl cells resulted in a remarkableincrease in levels of detectable AEC. This result indicatesthat conjugated AEC is synthesized and accumulated in the AEC-resistantcells. The level of the AEC conjugate in Bl cells increasedwith increases in the concentration of AEC in the culture medium,while intracellular levels of AEC were so low as not to be detectablein the case of cells grown on medium supplemented with AEC atless than 1 mM. The AEC conjugate was also detected in Adcells, but at lower levels than in the AEC-resistant cells.In addition, AEC was found to be incorporated into soluble proteinsin Ad cells. These results suggest that the resistance of AEC-resistant cellsof Datura innoxia is accomplished via acceleration of the synthesisof the AEC conjugate which prevents any increase in intracellularlevels of free AEC. 1Present address: Institute for Biology and Chemistry, TsumuraCo.Ltd., Inashiki, Ibaraki, 300-03 Japan. 2Present address: North Kanto Shop, Sakata Seed Co. Ltd.,Saitama,347 Japan.  相似文献   

2.
Growth kinetics of Datura innoxia batch suspension cultureswen monitored by a Klett-turbidimetric technique. While cultured. wt varied linearly with Klett units, f. wt and packed cellvolume did not. Turbidimetrically determined doubling timeswere highly reproducible. The method proved to be useful inthe determination of acutely lethal conantrations of a seriesof anti-metabolites. In certain circumstances, aggregate dissociation in batch suspensioncultures of D. innoxia was found to be coupled to growth rate.Suspensions maintained with 10–5 M 2,4-D exhibited a relativelyslow growth rate with a high degree of aggregate dissociation:10–4 M 2,4-D promoted a maximum growth rate, but dramaticallysuppressed aggregate dissociation. At 10–5 M 2,4-D, themitotic index of smaller-aggregate fractions was greater thanthe mitotic index of the large-aggregate fraction. At 10–5M 2,4-D the converse was observed. Supraoptimal 2,4-D concentrationsthus enhanced both aggregate dissociation and the growth ofsmaller aggregates. When present in concentrations promoting optimal growth. malicand succinic acids caused a decrease in aggregate dissociation.Casein hydrolysate dramatically enhanced growth, but did notaffect aggregate dissociation to the same degree as 2,4-D orthe Krebs cycle organic acids. Suggestions are made concerningmedium composition to be used in future mutant selection schemesusing D. innoxia. Datura innoxia (Miller), suspension culture, growth kinetics, mitotic index, 2,4-dichorophenoxy acetic acid  相似文献   

3.
Datura plants were grown on a clayed support and subjected tosalinity stress (153.8 mol m–3 NaCl) at the 6-leaf stage.Salt treatment increased total alkaloid content in young leaves.The results indicated that at the organ level tropane alkaloidaccumulation was related to plant growth. Key words: Datura innoxia, salt stress, tropane alkaloids  相似文献   

4.
Cells of the auxotrophic mutant, Ad1, of Datura innoxia requiredadenine, adenosine, or inosine for their growth on solid agarmedium which contained Murashige-Skoog salts, 2,4-dichloro-phenoxyaceticacid, and sucrose. Thirteen purine and pyrimidine nucleotidesin extracts of wild-type and Ad1 cells were separated and quantifiedby HPLC. Levels of ADP-glucose and UMP were significantly higherin Ad1 than in wild-type cells, but those of other nucleotideswas found when Ad1 cells were transferred to fresh medium withoutadenine. The rate of the biosynthesis de novo of purines, asestimated from the rate of incorporation of 14C from [2-14C]-glycine and [14C]formate into adenine nucleotides, was reducedin Ad1 cells to 21 and 13% of the wild-type rate, respectively.The activities involved in the salvage of adenine and adenosinein Ad1 cells were similar to those in wild-type cells. Ad1 cellshad the capability to convert adenine to guanine nucleotidesand guanine to adenine nucleotides. 1 Part 27 of the series, "Metabolic Regulation in Plant CellCulture". (Received March 7, 1988; Accepted August 3, 1988)  相似文献   

5.
The sensitivity of twelve strains of Saccharomyces cerevisiaeto Cd2+ was examined in correlation with the uptake of Cd2+.Strains of S. cerevisiae were grouped into three categoriesdepending on the sensitivity of cells grown on agar-plates containingvarious concentrations of Cd2+. 1) The sensitive group did notgrow in 0.1 mM Cd2+. 2) The sub-tolerant group was capable ofgrowth at 0.3 min Cd2+, but not at 0.4 mM Cd2+. 3) The tolerantgroup was capable of growth at 0.4 mM Cd2+ or higher. In thesestrain groups the increase in sensitivity to Cd2+ was associatedwith an increase in the activity of Cd2+ absorption. 1 This study is dedicated to the late president J. Ashida ofEhime University. (Received November 25, 1982; Accepted February 14, 1983)  相似文献   

6.
The green alga Kirchneriella lunaris was incubated with variousheavy metals (Cd2+, Co2+, Mn2+, Ni2+) in presence/absence ofcalcium (Ca2+). The uptake of heavy metal was affected by Ca2+.Growth rate was inhibited by all heavy metals applied. In allCa2+-containing cultures Kirchneriella exhibited higher ratesof growth than those containing heavy metal alone. Photosynthesis/respirationratio of K. lunaris cells seems to be the determinant in thiswork. Ca2+ variably abolished the effects of the heavy metalsstudied. Maximal positive effect of Ca2+ was found with Cd2+while with Ni2+ it was negligible.Copyright 1995, 1999 AcademicPress Cadmium, cobalt, manganese, nickel, calcium, heavy metals, growth, photosynthesis, Kirchneriella lunaris  相似文献   

7.
8.
重金属递进胁迫对黑麦草初期生长的影响   总被引:14,自引:0,他引:14  
通过研究Cu2+、Zn2+、Cd2+与Pb2+胁迫对黑麦草初期生长的影响,结果表明:4种重金属对种子发芽率抑制效应相对较小,尤其Cu2+与Zn2+的抑制作用最小。高浓度Cu2+、Cd2+胁迫对株高、根系长度、地上生物量的抑制作用相对较大,尤其Cu2+对根系生长的抑制效应最大,在300 mg·L-1下,与对照相比,根长最高下降了 85.48%。高浓度Cd2+胁迫显著降低了叶绿素含量,在300 mg·L-1时比对照降低了45.51%;与对照相比,Cu2+与Zn2+所有处理都增加了叶绿素含量。从递进胁迫进程看,一些重金属对某一生长指标的影响往往表现在低浓度具有促进作用,而高浓度又存在明显的抑制效应。  相似文献   

9.
A Cd-resistant strain of yeast (Saccharomyces cerevisiae, strain30IN) accumulated Cd with the concomitant synthesis of a Cd-bindingprotein of low molecular weight when grown in Cd2+-containingmedium. Analysis of the amino acid composition, N-terminal sequenceand immunological properties of the protein revealed its structuralhomology to Cu-metallothionein (Cu-MT) in S. cerevisiae 2186,a Cu-resistant strain of yeast (Winge et al. 1985). The synthesisof MT in Cu-resistant strains of yeast is known to be underthe strict control of Cu2+ ions, while that in 301N was inducedboth by Cd2+ and Cu2+ ions. When 301N was precultured for 48h in the presence of 1 mM CuSO4, its resistance to Cu2+ andthe synthesis of MT in response to Cu2+ were enhanced whileanalogous responses to Cd2+ were conversely reduced. These resultssuggest that the synthesis of MT is controlled by Cd2+ and Cu2+in a counteractive manner in strain 301N and, therefore, theregulation of the synthesis of MT plays a role in the adaptationof this strain to conditions when either metal is present. (Received November 1, 1990; Accepted February 22, 1991)  相似文献   

10.
Shikimate kinase from Phaseolus mungo seedlings was partiallypurified by DEAEcellulose, hydroxyapatite and Sephacryl S-200column chromatographies. The activity was completely inhibitedby EDTA and the requirement for Mg2+ could be partially replacedby Mn2+, Ca2+; Co2+ and Cd2+. Sulfhydryl inhibitor did not inhibitthe enzyme activity. The apparent Km values for shikimic acidand ATP at pH 8.6 were 0.25 mM and 0.38 mM, respectively. Theactivity appeared to be maximal at pH 8.6–9.0. Shikimate-3-phosphateand ADP inhibited the activity slightly. Aromatic amino acids,quinic acid and dehydroquinic acid had no significant effecton the activity. (Received January 11, 1979; )  相似文献   

11.
A major target of cadmium (Cd2+) toxicity is the kidney proximal tubule (PT) cell. Cd2+-induced apoptosis of PT cells is mediated by sequential activation of calpains at 3–6 h and caspases-9 and -3 after 24-h exposure. Calpains also partly contribute to caspase activation, which emphasizes the importance of calpains for PT apoptosis by Cd2+. Upstream processes underlying Cd2+-induced calpain activation remain unclear. We describe for the first time that 10–50 µM Cd2+ causes a significant increase in ceramide formation by 22% (3 h) and 72% (24 h), as measured by diacylglycerol kinase assay. Inhibition of ceramide synthase with fumonisin B1 (3 µM) prevents ceramide formation at 3 h and abolishes calpain activation at 6 h, which is associated with significant attenuation of apoptosis at 3–6 h with Hoechst 33342 nuclear staining and/or 3(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) death assays. This indicates that Cd2+ enhances de novo ceramide synthesis and that calpains are a downstream target of ceramides in apoptosis execution. Moreover, addition of C6-ceramide to PT cells increases cytosolic Ca2+ and activates calpains. Apoptosis mediated by C6-ceramide at 24 h is significantly reduced by caspase-3 inhibition, which supports cross talk between calpain- and caspase-dependent apoptotic pathways. We conclude that Cd2+-induced apoptosis of PT cells entails endogenous ceramide elevation and subsequent Ca2+-dependent calpain activation, which propagates kidney damage by Cd2+. nephrotoxicity; cell signaling; cell biology and structure  相似文献   

12.
The effect of treatment of wheat plants with Cd2+ ions on thephotochemical activity of the primary leaves was examined. Threeday-old etiolated seedlings were treated with Cd2+ ions for24 h in dark, and after this treatment the plants were grownin the light until the primary leaves were fully developed.Cd2+ ions (30–120 µM) induced a significant decreasein activities of both photosystem II and photosystem I. Theextent of the decrease in PS II activity was much greater thanthat in the PS I activity. Analysis of changes in the fluorescenceyield of chlorophyll also indicated that Cd2+ ions drasticallyaffect the photochemistry of photosystem II. Cd2+ ions induceddecrease in the rates of photoreduction of 2,6-dichlorophenolindophenol even in the presence of the exogenous electron donor,hydroxylamine, both in Tris-treated and untreated chloroplasts.This result suggests that the site of inhibition is near thesite of donation of electrons by hydroxylamine. Treatment withCd2+ ions impairs the electron transport system on the reducingside of PS II. The decrease in the fluorescence yield of Chi is less than that in the evolution of O2 mediated by oxidizedphenylenediamine. This difference may be a result of inhibitionon the reducing side of PS II. In addition to inhibition onthe reducing side, Cd2+ ions may affect the oxidizing side ofPS II. A comparative study of the rates of evolution of O2 withp-benzoquinone and dichloro-p-benzoquinone as electron acceptorswas performed since the halogenated benzoquinones have beenshown to accept electrons from both active and inactive centersof photosystem II while some of the benzoquinones accept electronsonly from active centers. The results suggest that Cd2+ ionsinduced a marginal increase in the number of inactive reactioncenters in PS II. Analysis of light-saturation-kinetics of theevolution of O2 catalysed by PS II indicates a reduction inthe size of the antennae as well as in the concentration ofthe active (-type) reaction centers of PS II. Thus, the Cd2+-inducedeffects on the photochemistry of PS II involve changes on thereducing side of PS II as well as possible changes in the sizesof the populations of active and inactive centers. Thus, short-termexposure to Cd2+ ions during establishment of seedlings hasa severely detrimental effect on photochemical activities inchloroplasts. (Received October 17, 1990; Accepted July 3, 1991)  相似文献   

13.
The distribution and accumulation of Cu2+, Zn2+ and Cd2+ ionsin the roots of Polygonum cuspidatum, collected from metalliferousand non-metalliferous habitats, were examined. About 90% ofthe metal ions was located in the cell wall fraction of rootsof plants grown in either type of habitat. The metal-ion exchangecapacity of the cell walls is not specific to the plants frommetalliferous habitats, and there were no significant differencesamong plants from the various habitats. The capacity for Cu2+ions was greater than that for Zn2+ or Cd2+ ions. Cu2+ ionshave a high affinity for the cell wall and, thus, it appearsthat the cell wall is a barrier for the transportation of Cu2+into the cytoplasm. (Received March 10, 1988; Accepted March 17, 1989)  相似文献   

14.
Sugar beet (Beta vulgaris L. cv. Monohill) were cultivated ina nutrient solution with different combinations of Ca2+ (36,180, 720 or 3560µM) and Cd2+ (0, 1, 5 or 20µM).The dry and fresh weights, the content of Ca2+ and Cd2+ , sucrose,fructose, glucose and starch in 5-week-old plants was analysedas well as the rate of [14C]-sucrose uptake in discs from 3-month-oldstorage roots. The carbohydrate metabolism was indirectly affectedby the presence of calcium or cadmium. Cadmium caused a diminisheddry weight and carbohydrate concentration. The dry weight wasunaffected by the Ca2+ level but the carbohydrate distributionbetween storage and growth processes was affected; at low Ca2+in the tissue, the growth was retarded and the level of storagecarbohydrate increased, while at high Ca2+ the opposite wasfound. The [14C]-sucrose uptake decreased in tap roots cultivatedat low Ca2+ . Long term exposure to Cd2+ also decreased thesucrose uptake in tap roots. Direct Cd2+ addition to the assaymedium, however, increased the sucrose uptake, probably at thetonoplast, while Ca2+ had no transient effect on the uptake.Cadmium increased the Ca2+ concentration in the plant, but Ca2+did not affect the net-uptake of Cd2+. Key words: Sugar beet, cadmium uptake, calcium uptake, carbohydrate formation, growth  相似文献   

15.
Amakawa  Taisaku 《Chemical senses》1978,3(4):413-422
1) Ca+ + (1 to 10 mM) lowered the binding affinity of sugarreceptor-site for sucrose in the labellar sugar receptor ofthe blowfly, Phormia regina, without changing the maximum-responseamplitude. It also elevated the values of the Hill coefficient(nH) in some degrees. 2) Other divalent cations such as Mg+ +, Ba+ + or Cd+ + alsoshowed almost the same property as above. The sequence of theeffect is as follows: Ba+ +, Mg+ + x Ca+ + x Cd+ +. Trivalentcation, La+ + + (1 mM), changed the value of nH from 1 (La++ +-free) to 2. 3) On the contrary, the action of monovalent cations such asK+ or Na+, of which ionic strength was made the same as thatof the divalents hardly suppressed the response. 4) The results obtained do not support the hypothesis, at leaston the sugar receptor of the fly, that the receptor potentialis attributable to a change of the surface potential (zeta potential)as is proposed for the frog sugar receptor.  相似文献   

16.
The mechanism involved inN-methyl-D-glucamine(NMDA)-induced Ca2+-dependentintracellular acidosis is not clear. In this study, we investigated indetail several possible mechanisms using cultured rat cerebellargranule cells and microfluorometry [fura 2-AM or 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein-AM].When 100 µM NMDA or 40 mM KCl was added, a marked increase in theintracellular Ca2+ concentration([Ca2+]i)and a decrease in the intracellular pH were seen. Acidosis wascompletely prevented by the use ofCa2+-free medium or1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-AM, suggesting that it resulted from an influx of extracellular Ca2+. The following fourmechanisms that could conceivably have been involved were excluded:1)Ca2+ displacement of intracellularH+ from common binding sites;2) activation of an acid loader or inhibition of acid extruders; 3)overproduction of CO2 or lactate; and 4) collapse of the mitochondrialmembrane potential due to Ca2+uptake, resulting in inhibition of cytosolicH+ uptake. However,NMDA/KCl-induced acidosis was largely prevented by glycolyticinhibitors (iodoacetate or deoxyglucose in glucose-free medium) or byinhibitors of the Ca2+-ATPase(i.e.,Ca2+/H+exchanger), including La3+,orthovanadate, eosin B, or an extracellular pH of 8.5. Our results therefore suggest that Ca2+-ATPaseis involved in NMDA-induced intracellular acidosis in granule cells. Wealso provide new evidence that NMDA-evoked intracellular acidosisprobably serves as a negative feedback signal, probably with theacidification itself inhibiting the NMDA-induced[Ca2+]i increase.

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17.
Wolterbeek, H. Th. 1987. Relationships between adsorption, chemicalstate and fluxes of cadmium applied as Cd(NO3)2 in isolatedxylem cell walls of tomato.—J. exp. Bot. 38: 419–432. Isolated xylem cell wall pieces were applied as membranes inion diffusion experiments. The cell walls were isolated fromtomato internodes (Lycopersicon esculentum Mill, cv. Tiny Tim)and sealed in a two-compartment diffusion system. In flux andadsorption calculations, the cell wall was regarded as a leakymembrane with parallel fluxes through Donnan Free Space (DFS)and Water Free Space (WFS). During the experiments absorptioninto and diffusion across the walls was determined of Cd2 +, applied as 115Cd(NO3)2. Flux experiments with 82Brindicated that excluded volume effects and path tortuosity resultedin apparent WFS diffusion coefficients in the walls which were0·012 times as high as in water. The free proton concentration in the DFS was shown to be relatedto a complex formation between fixed charges and Cd2 +. Thecell wall permeability for Cd2 + and NO3 varied withapplied and absorbed concentrations, and the Cd2 + flux curveshowed an inflexion point coinciding with a buffered degreeof dissociation of fixed charges in the DFS. The necessary couplingof fluxes of opposite charges resulted in relatively high NO3and small Cd2 + permeability of the DFS for strongly dilutedsolutions (P = 10–4 m s–1 and 10–11 m s–1for NO3 and Cd2 + respectively). The results demonstratethe possible regulatory effects of the cell wall in processesof ion transfer from xylem vessels, or ion uptake in plant tissues. Key words: Cadmium, chemical state, DFS, WFS, ion flux, permeability, xylem cell walls, tomato, bromium, nitrate  相似文献   

18.
 以耐铝性明显差异的两个大豆(Glycine max)基因型‘浙秋2号’(耐性)和‘浙春3号’(敏感)为材料, 研究根尖边缘细胞比活度、粘液分泌和根长对铝胁迫和解除胁迫的反应, 明确边缘细胞的粘液分泌对策在铝毒环境中的生态学意义。结果表明, ‘浙秋2号’在100~400 µmol&;#8226;L–1 Al3+处理的3~12 h, 边缘细胞比活率呈递减趋势, 12 h后比活率又略有上升。‘浙春3号’在300和400 µmol&;#8226;L–1 Al3+处理的变化与前者一致。两个大豆基因型的粘液层随着Al3+浓度增加和时间延长而增厚, 并于400 µmol&;#8226;L–1 Al3+处理24 h时达到最大(>17 µm)。‘浙秋2号’在低浓度Al3+ (100和200 µmol&;#8226;L–1)处理3~6 h后就会分泌大量粘液, ‘浙春3号’则在300 µmol&;#8226;L–1 Al3+处理12 h后才有类似的变化。‘浙秋2号’在400 µmol&;#8226;L–1 Al3+处理下的根相对伸长率均高于100~300 µmol&;#8226;L–1 Al3+处理, ‘浙春3号’则表现为Al3+浓度越高, 根伸长受抑越明显。Al3+胁迫解除后, ‘浙秋2号’的粘液分泌速度和分泌量急剧下降, ‘浙春3号’在胁迫解除后的24 h, 仍会持续、大量地分泌粘液(>19 µm)。可见, 耐性大豆通过在铝胁迫初期快速、大量地分泌粘液以维持较高的边缘细胞活性和解除胁迫后迅速降低粘液的分泌速度及分泌量来适应铝毒害环境。  相似文献   

19.
In manynonexcitable cells, hormones and neurotransmitters activateNa+ influx and mobilizeCa2+ from intracellular stores.The stores are replenished by Ca2+influx via "store-operated"Ca2+ channels (SOC). The mainroutes of Na+ entry in these cellsare unresolved, and no role forNa+ in signaling has beenrecognized. We demonstrate that the SOC are a majorNa+ entry route in arterialmyocytes. Unloading of the Ca2+stores with cyclopiazonic acid (a sarcoplasmic reticulumCa2+ pump inhibitor) and caffeineinduces a large externalNa+-dependent rise in thecytosolic Na+ concentration. Onecomponent of this rise in cytosolicNa+ concentration is likely due toNa+/Ca2+exchange; it depends on elevation of cytosolicCa2+ and is insensitive to 10 mMMg2+ and 10 µMLa3+. Another component isinhibited by Mg2+ andLa3+, blockers of SOC; thiscomponent persists in cells preloaded with1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraaceticacid to buffer Ca2+ transients andpreventNa+/Ca2+exchange-mediated Na+ entry. ThisNa+ entry apparently is mediatedby SOC. The Na+ entry influencesNa+ pump activity andNa+/Ca2+exchange and has unexpectedly large effects on cell-wideCa2+ signaling. The SOC pathwaymay be a general mechanism by which Na+ participates in signaling inmany types of cells.

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20.
Sugar beet seedlings (Beta vulgaris L. cv. Monohill) were grownfor 14 d on a nutrient solution based on the nutrient proportionsin healthy plants. Nutrients were supplied either once at relativelyhigh concentrations, or in small amounts with a daily incrementalincrease of 0?15 or 0?20 in accordance with an exponential growthrate. Cadmium (0, 0?6, 2?3, 50 or 20?0 µmol) was introducedeither by a single addition or in daily increments of 0?15 or0?20. Cadmium uptake, expressed as a percentage of total Cd2+supplied, decreased with increasing total Cd2+ content and withdecreasing availability of nutrients. With a daily supply ofcadmium, net uptake, transport and content per unit of dry weightin roots and shoots were related to the total Cd2+ supplied.Cadmium caused growth retardation, increased root/whole-plantratio, and decreased root-tip respiration and photosynthesis.At high initial nutrient concentrations, Cd2+ decreased thecontents of sucrose, glucose, fructose, and starch per unitof dry weight. The opposite was found if nutrients were addeddaily. In the latter case, the dry weight/fresh weight ratioalso increased. The effects of cadmium were related to [Cd2+]in proportion both to the root absorption area and to the nutrientconcentration. Key words: Sugar beet, mineral provision, cadmium uptake, sugar formation, growth  相似文献   

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