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1.
Changes in the activities of some glycosidases were studiedin carrot suspension cultures with and without 2,4-D. Remarkablecell elongation occurs in a medium without 2,4-D, while fewcells elongate in a medium containing it. Glycosidases werefractionated into soluble, ionically wall-bound, tightly wall-boundand extracellular enzymes. The optimum pHs of all the ionicallybound glycosidases were in an acidic range, 4.4–5.0. The activities of the ionically and tightly bound ß-xylosidasesand ß-galactosidases were higher in elongating thanin non-elongating cells. Furthermore, the activities of theseenzymes increased with cell elongation during culture, suggestingthat they may play important roles in cell elongation. Higheractivities of soluble and cell wall-bound ß-glucosidaseand -mannosidase were found in non-elongating rather than inelongating cells. The activities of all soluble glycosidasesexcept ß-xylosidase were also higher in non-elongatingcells. Only ß-xylosidase and ß-galactosidaseactivities were detectable in the medium of the elongation culture. 1 Present address: Department of Agricultural Chemistry, ObihiroUniversity of Agriculture and Veterinary Medicine, Obihiro,Hokkaido 080, Japan.  相似文献   

2.
Gas chromatography was used to measure ethylene (ethene) andethane production by tobacco (Nicotiana tabacum cv. Wisconsinno. 38) callus tissues grown on media containing inorganic saltsaccording to Murashige and Skoog (1962), sucrose, myo-inositol,thiamine-HCl kinetic according to Linsmaier and Skoog (1965),and either 2,4-dichiorophenoxyacetic acid (2,4-D) in the range0–100 mgl–1 or 2 mgl–1 indoi-3-ylacetic acidplus NaCl in the range 0–200 Meq l–1. Ethylene productionrates were high (> 500 nl h–1 g1– fresh weight)initially in all treatments. Subsequently, ethylene productiondeclined in rapidly growing cultures but remained high in moderatelyand severely 2,4-D (> 0·5 mgl–1) stressed andin severely NaCl (150 Meql–1) stressed cultures. Highinitial rates of ethane production (> 200 nl h–1 g–1fresh weight) were obtained under conditions of severe stresscaused by 2,4-D or NaCl but not in control or moderately inhibitedcultures. With further incubation ethane production declinedin the severely stressed cultures. It is concluded that ethyleneproduction can be used as an index of moderate 2,4-D stressand severe NaCl stress by virtue of the high persisting ratesof ethylene production in stressed cultures. Ethane productioncan be used as an early index of severe stress caused by either2,4-D or NaCl in vitro. Nicotiana tabacum L., tobacco, ethylene, ethenen, ethane, 2,4-dichlorophenoxyacetic acid, auxin, stress, callus tissue  相似文献   

3.
Adventitious plantlets were obtained from lateral buds, shoottips, embryos, and pieces of stem and rachilla tissue of Phoenixdactylifera L. cultured on a modified Murashige and Skoog mediumcontaining 3 mg l–1 N-(2-isopenty)adenine 0?1–100mg l–1 -naphthaleneacetic acid or 2,4-dichlorophenoxyaceticacid, and 3 g l–1 activated charcoal. Additions of auxinswere necessary to induce explants to produce callus, adventitiousplantlets, and roots. Plantlets were obtained from explantscultured 3–4 months in vitro. No difference in growthresponses between male and female explants was observed duringculture. Complex addenda of activated charcoal and polyvinylpyrrolidonewere tested in the nutrient media at various concentrationsto prevent explant browning. Activated charcoal fostered satisfactorygrowth by reducing the browning and inhibition of growth ofexplants.  相似文献   

4.
Fidgeon, C. and Wilson, G. 1987. Growth regulation of Galiummollugo L. cell suspensions by -naphthalene acetic acid.—J.exp. Bot. 38: 1491–1500. Galium mollugo cell suspension cultures were found to requirethe plant growth regulator -naphthalene acetic acid (-NAA) forcontinued growth and cell division. This requirement could notbe substituted in either batch or semi-continuous culture byindole-3-acetic acid (IAA) or 2,4-dichlorophenoxy acetic acid(2,4-D) at any concentration tested. However, ß-naphthaleneacetic acid (ß-NAA) and indole-3-butyric acid (IBA)were found to support growth when supplied at a concentrationtwo orders of magnitude greater than the normal media level(0–5 mg dm3). The growth of Galium cells was found to be influenced not onlyby the -NAA initially supplied in the medium but also by theexposure to -NAA in previous growth cycles. Preculture of cellsfor 3 d in an -NAA containing medium, followed by cell washingand re-inoculation into -NAA free medium, supported a quantitativegrowth response similar to that obtained after 14 d in the control-NAA containing medium. Even short-term exposures between 0·5and 6·0 h stimulated a detectable growth response 14d later. These observations raise questions relating to theuptake and perception of exogenously supplied growth regulatorsby cultured cells. The delayed kinetics of this form of response is of significancein culture regimes in which cells are transferred from one mediumto another, differing in their growth regulator composition,in order to induce morphogenesis  相似文献   

5.
-Aminobutyric acid (GABA) accumulation occurs in cultured ricecells when ammonium is added to the medium [Kishinami and Ojima(1980) Plant Cell Physiol. 21: 581–589]. Whether thisphenomenon occurs in rice plant tissues was examined with respectto exogenously supplied auxins: 2,4-dichlorophenoxyacetic acid(2,4-D), indole-3-acetic acid (IAA) and naphthalene-acetic acid(NAA). In intact rice plants grown in medium containing ammonium withoutauxin, glutamine first increased, then asparagine graduallyincreased. In both shoots and roots, the asparagine contentbecame the highest among four amino acids after 4 days of cultureperiod. GABA did not increase at all, its level remaining lowin both shoots and roots throughout the culture period. GABA accumulation was observed in excised rice root tips whenthey were incubated in the medium containing ammonium in thepresence of 2,4-D, IAA or NAA. In the absence of auxin, however,excised rice root tips accumulated asparagine and glutamine,but not GABA. Rice root segments obtained from a region in whichroot cells had already developed to maturity did not accumulateGABA but asparagine and glutamine in the presence of both ammoniumand 2,4-D. These results suggest that GABA accumulation occurs in rapidlygrowing and dividing tissue, such as the apical meristem ofrice root in the presence of auxin during ammonium assimilation. (Received June 15, 1987; Accepted March 14, 1988)  相似文献   

6.
CALOIN  M.; YU  O. 《Annals of botany》1982,49(5):599-607
The kinetics of growth of Dactylis glomerata L. were studiedunder controlled temperature and nutritional conditions at threelevels of irradiance (35·55 and 85 W m–s). Thedry weights of the root and shoot parts of the plants were measuredeach week between the fourth and eleventh weeks after sowing. The growth kinetics were found to be dependent on the levelof irradiance, but no significant differences in the root: totaldry weight ratio were observed. To characterize the effect of illuminance, the experimentalgrowth curves were analysed initially using the logistic model,the adequacy of which is discussed. An extension of the logisticmodel is proposed, represented by the kinetic equation dM/dt= kM, with < 1 and where M is the dry weight of the plant.It is shown that this relationship allows a distinction to bemade between two kinds of plant material according to theirfunctions in the growth process. Dactylis glomerata L., illuminance, growth curves, kinetic analyses, logistic model, shoot:root ratio, partition of assimilates  相似文献   

7.
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   

8.
A Ras-related NTP-binding protein was partially purified froma membrane fraction derived from the mycelia of Neurospora crassa.[-32P]ATP and [-32P]GTP were incubated with mem brane and solublefractions which were then irradiated with UV light to inducecrosslinking of tightly bound nucleotides. After SDS-polyacrylamidegel electrophoresis, blotting onto a nitrocellulose filter andautoradiography it was apparent that most of the proteins thatbound [-32P]-GTP also bound [-32P]ATP. Pretreatment of the membranefraction with Ras-specific antibody effectively blocked thebinding of [-32P]ATP and [-32P]GTP to several ATP-GTP-bindingproteins. The band of a protein with a molecular weight of 26kDa on the SDS-polyacrylamide gel cross-reacted strongly withthe Ras-specific antibody. The protein was extracted from thegel and further purified by repeated gel electrophoresis. Thepurified protein bound [-32P]ATP, [-32P]-GTP, [-32P]CTP and[-32P]UTP at 1.6x10 M and was autophosphorylated in thepresence of [-32P]ATP and [-32P]GTP at 1.7x10 M. Pretreatmentof the protein with Ras-specific antibody partially blockedthe autophosphorylation in the presence of these nucleotides.The binding of [-32P]ATP to the NTP-binding protein was blockedby addition of ATP at 10–4–10–3 M. ATP ata concentration of 10–4 M prevented the binding of [-32P]to a greater extent than did GTP at the same concentration.Binding of [-32P]CTP and [-32P]UTP to the protein was also observed. (Received October 7, 1991; Accepted July 14, 1992)  相似文献   

9.
An 18-h treatment of synchronously-grown Chlorella pyrenoidosawith 2,4-D did not significantly alter the size, dry weight,degree of synchrony, or pigment content of the cells, nor weredetectable quantities of ethylene produced. When Chlorella pyrenoidosawas treated with 5?10–4 M 2,4-D, there was a statisticallysignificant stimulation of both net oxygen uptake and productionwhile 5?10 M 2,4-D inhibited both processes. When Chlorellapyrenoidosa was treated with 5?10–4 M and 5?10–3M 2,4-D, significantly greater amounts of glycollate were presentin the culture medium, even though an assay for glycollate dehydrogenaseshowed that the activity of this enzyme from 2,4-D-treated Chlorellapyrenoidosa was three times greater than in control cells. Looselybound 2,4-D was partitioned from a nonaqueously isolated chloroplastfraction, while other cell fractions failed to show detectablequantities of 2,4-D. It is postulated that in Chlorella pyrenoidosathe chloroplast is a target for 2,4-D action and that interferencein photorespiratory processes may underlie the observed responses.  相似文献   

10.
Transport of fluid by lens epithelium   总被引:2,自引:0,他引:2  
We report for the first time that cultured lens epithelial celllayers and rabbit lenses in vitro transport fluid. Layers of the TN4mouse cell line and bovine cell cultures were grown to confluence onpermeable membrane inserts. Fluid movement across cultured layers andexcised rabbit lenses was determined by volume clamp (37°C).Cultured layers transported fluid from their basal to their apicalsides against a pressure head of 3 cmH2O. Rates were (inµl · h1 · cm2)3.3 ± 0.3 for TN4 cells (n = 27) and 4.7 ± 1.0 for bovine layers (n = 6). Quinidine, a blocker ofK+ channels, andp-chloromercuribenzenesulfonate andHgCl2, inhibitors of aquaporins,inhibited fluid transport. Rabbit lenses transported fluid from theiranterior to their posterior sides against a2.5-cmH2O pressure head at 10.3 ± 0.62 µl · h1 · lens1(n = 5) and along the same pressurehead at 12.5 ± 1.1 µl · h1 · lens1(n = 6). We calculate that this flowcould wash the lens extracellular space by convection about once every2 h and therefore might contribute to lens homeostasis and transparency.  相似文献   

11.
The purpose of this study was to compare human endothelial cells from Schlemm's canal (SCEs) and the trabecular meshwork (TMEs) in terms of ZO-1 isoform expression, hydraulic conductivity (HC) properties, and "giant" vacuole (GV) formation. The principal study methods were Western blot, RT-PCR, immunofluorescence, and perfusion chambers. Blot signals for +-and --isoforms were similar in SCEs but less intense for the +-relative to the --signal in TMEs. With the anti-+ antibody used at 1/50 dilution, binding occurred at cell borders of both cell types, but only to SCEs when used at a =" BORDER="0">1/200 dilution in vitro and in vivo. SCEs were more resistive than TMEs (HC = 0.66 vs. 1.32 µl·min-1·mmHg-1·cm-2; P < 0.001) when perfused from apex to base. When perfused in the other direction, SCEs were again more resistive (5.23 vs. 9.04 µl·min-1·mmHg-1·cm-2; P < 0.01). GV formation occurred only in SCEs as a function of flow direction, perfusion pressure, and time. We conclude that SCEs and TMEs have distinctive phenotypic properties involving their content of ZO-1 isoforms, barrier function, and GV formation. tight junctions; ZO-1; giant vacuoles; conductivity  相似文献   

12.
Young leaf segments of Zea mays L. seedlings were cultured onMurashige and Skoog's basal nutrient medium supplemented with2 mg l–1 2, 4-D and sub-cultured on medium containing8 mg l–1 2,4-D. Two types of callus tissues appeared—embryogenicand non-embryogenic. The embryogenic callus tissue producednumerous somatic embryos which on transfer to media containinglow amounts of 2,4-D or ABA produced plantlets. Callus tissuesexhibited embryogenic potential for more than 1 year. Zea mays L. cv. Ageti-76, Zea mays L. cv. N-L-D-Comp., maize, leaf, callus, somatic embryogenesis, regeneration  相似文献   

13.
The effect of substituted pyridines on the response of singlepyridine-sensitive cells from crayfish walking legs was investigatedelectrophysiologically. Seven p-substituted pyridines, actingreversibly, were identified as specific antagonists at the pyridinereceptor. The maximum saturation frequency of the response toagonists was reached in the presence of antagonists but thedose–response curves of the agonists were shifted in parallelto the right along the concentration axis. Schild plots of threehighly effective antagonists with six agonists were linear witha slope close to one, indicating competitive antagonism. Theinhibition constants yielded a K1 value of {small tilde}4–8µmol for the most effective antagonist, 4-(4-nitrobenzyl)pyridine,which had only one order of magnitude lower affinity than themost effective agonist 2,3'-bipyridyl. The antagonists 2,4'-bipyridyland 4-benzylpyridine had K1 values of 6–10 µmol,followed by 4-acetylpyridine with a K1 value of 30–70µmol. The rank order of inhibitory potency of the differentantagonists was found to be the same for all units tested. Comparingelectronic effects (Hammett values and pKa values) of the substitutentsin p- and m-position showed that inhibitory effectiveness decreasedwith a decrease in pKa and an increase in Partition coefficientswere determined for 10 agonists and the antagonists which weregenerally more lipophilic than the agonists. A hypotheticalreceptor site is proposed.  相似文献   

14.
Primary production was measured for 7 years, using the in situ14C-method in hypertrophic Hartbeespoort Dam, South Africa,to examine the influence of light and water temperature on theupper limit of Microcystis aeruginosa production. Water temperaturesvaried from 11 to >25°C and chlorophyll concentrationsreached 6500 mg m–3. The maximum volumetric rate of production(Amax) was 12->8800 mg C m–3 h–1 with areal productions(A) of 69->3300 mg C m–2 h–1 for euphotic zonedepths of <0.5–8.4 m. The intrinsic parameters of phytoplanktonproduction (, Amax/B, Ik) indicated that the phytoplankton populationwas adapted to high light levels. Both Amax/B and Ik were correlatedwith temperature. Under optimal conditions, , the theoreticalupper limit of A, was calculated to be 2.8 g Cm–2 h–1,while the measured rate was 2.5 g Cm–2 h–1. Measuredareal rates exceeding were overestimated due to methodologicalproblems when working with Microcystis scums. Light and watertemperature interacted to yield high production rates: watertemperature through its direct effect on photosynthetic ratesand indirectly in the formation of diurnal mixed layers; lightindirectly through water temperature and directly through itsattenuation and induction of light-adapted physiology in Microcystis.  相似文献   

15.
STAMP  J. A. 《Annals of botany》1987,59(4):451-459
Anatomical and morphological studies demonstrated that somaticembryos developed similarly on mature seed and clonal leaf explantsof cassava (Manihot esculenta Crantz) cultured for 20–24d on Murashige and Skoog (MS2) basal medium supplemented with4.0 mg l–1 2,4-D (Stage 1) before transfer to MS2 basalmedium supplemented with 0–01 mg l–1 2,4-D and 0–1mg l–1 6-benzylaminopurine (Stage II medium). Within 7d of inoculation onto Stage I medium, cell divisions occurredin the adaxial tissues of cotyledon-piece and leaf-lobe explants,and associated with this was the development of embryogeneticprotusions and ridges on the adaxial surface. Foliose structuresand somatic embryo initials developed from these tissues oncotyledon, embryonic axis and leaf-lobe explants and, when cultureswere transferred to Stage II medium, further somatic embryodevelopment occurred. Somatic embryos apparently originatedfrom groups of cells and were identified by the presence ofa closed root axis, a shoot axis and cotyledons of similar shapeand venation to those of zygotic embryos. Somatic embryos hadno vascular connection with parental cultures. Manihot esculenta, cassava, somatic embryogenesis, tissue culture, anatomy, morphology, morphogenesis  相似文献   

16.
Effects of respiratory inhibitors on frond and flower productionin light culture of a long-day duckweed, Lemna gibba G 3, wereinvestigated. The inhibitors examined could be divided into3 groups based on their specific actions: (A) 2,4-Dinitrophenol(10–6M), arsenate (10–4M), malonate (10–2M),o-phenanthroline (10–6M), ,'-dipyridyl (10–5M) andazide (10–6M) inhibited flower production by suppressingthe rate of flower production without affecting the inductionperiod. Frond production, however, was promoted by these reagents.Effective time of application came one day after the end ofthe induction period. (B) Iodoacetate (10–6M) and fluoride(10–4M) inhibited both flower production and, less significantly,frond production. Reduced rate of flower production was responsiblefor the inhibition of flowering. Effective time of applicationpreceded by one day that of A group inhibitors. (C) Salicylaldoxime(10–6M), diethyldithiocarbamate (10–6M) and 8-hydroxyquinoline(10–7M) enhanced flower production by reducing the lengthof the induction period, and simultaneously slightly inhibitedfrond production. Effective time of application was the latterhalf of the induction period. The implications of these findingsare discussed with special reference to the component processesinvolved in photoperiodic induction of flowering in duckweed. (Received March 27, 1969; )  相似文献   

17.
Effects of light flux density (LFD) during growth and uptakeassay on induction of transport system and kinetics of transport were studied using the Azolla pinnata-Anabaena azollae association (Azolla). Theinduction and uptake kinetics of the transport system were determined using an automated system that measuredthe NO3 concentration in the growth medium as a function oftime, using an on-line high performance liquid chromatograph(HPLC) with a UV-VIS detector. Full induction of the transport system required about 1.5 to 2.0 h and occurred without any apparent lag phase,regardless of the LFD provided. The level of induction of transport of Azolla grown at 600 µmol m–2s–1 LFD was higher than for that grown at 100 µmolm–2 s–1. Similarly, 600 µmol m–1 s–1LFD during the assay resulted in a higher level of inductionthan did 100 umol m–2 s–1. An increase in the LFDeither during the growth or the assay period increased the uptake rate; however, an increase in LFD duringthe latter period had greater effect. Azolla grown and assayedat 600 umol m–2 s–1 had the highest uptake rate. The uptake rate at 50 cm3 m–3ambient CO2 concentration was initially higher than at 305 cm3m–3, but the uptake rate decreased rapidly with time andeventually dropped below that at 305 cm3 m–3 CO2. Thesedata suggest that the energy required for transport in Azolla may bypass the photosynthetic CO2 fixationand carbon-cycling. Key words: carbon dioxide, concentration dependence, light flux density, uptake  相似文献   

18.
The development of taurine uptake into the unicellular greenalga Chlorella fusca 211-8b was characterized as a specificresponse to either nitrate or sulphate limitation. Taurine transportunder nitrogen starvation was stimulated by low pH and showeda biphasic kinetics with Km-values of 1.1 x 10–3 mol dm–3and 1.0 x 10–2 mol dm–3. Uptake was substantiallyinhibited by all - and ß-amino acids tested, whereassulphonate analogues failed to diminish taurine accumulation.Thus, uptake seemed to be mediated by a ‘general aminoacid permease’, unable to discriminate between carboxyland sulphonyl groups. However, Chlorella fusca could not catabolizethis unusual ß-amino acid and mobilize the amino-boundnitrogen for growth. Only a small group of -amino acids supportedthe growth of Chlorella fusca as an efficient nitrogen source. Key words: Taurine uptake, nitrogen starvation, amino acid uptake, Chlorella fusca.  相似文献   

19.
Smith, J. R., Smith, F. A. and Walker, N. A. 1987. Potassiumtransport across the membrane of Chara. I. The relationshipbetween radioactive tracer influx and electrical conductance.—J.exp. Bot. 38:731–751. The 42K influx () and the electrical conductance (Gm) were measured simultaneously for the ‘membrane’of internodal cells of Chara australis as a function of theexternal [KCl] (K?. In bathing solutions of pH = 5?0, progressively increased from 20?5to 430?60 nmol m–2 s–1 and Gm increased from 0?36?0?02to 3?8?0?8 S m–2 when K? was increased from 0?1 to 10mol m–3. The resting membrane potential difference (p.d.)was approximately -135 mV for low K? and approached the expectedNernst equilibrium p.d. for K+ ions when K? > 1?0 mol m–3.Measurements of 36Cl influx suggested that the 42K influx waspredominantly electrogenic. The equivalent Goldman permeabilityto K+ ions (Pk) was approximately 20–30 nm s–1 anddid not vary significantly with increasing K?. The equivalentconductance attributable to the electrogenic transport of K+ ions was calculated from assuming passive, independent diffusionof K+ ions and the ratio was found to be typically close to one. It was also found that themagnitudes of and Gm measuredsimultaneously for each individual cell were also well correlatedfor K? 1?0 mol m–3, and that the slope of the line ofbest fit was close to one. For each K? it was found that theconductance not attributable to K+ translocation and presumablyassociated primarily with the transport of protons or theirequivalents was typically 0?2–0?5 Sm–2. For K? >1?0 mol m–3 the results indicated that the transport ofK+ ions was essentially independent, i.e. there was no evidencefor flux interactions. The results also indicated that the equivalentconductance derived from the measured 42K influx could usefullyindicate the fraction of the electrical conductance attributableto the translocation of K+ ions. Key words: Potassium, conductance, influx  相似文献   

20.
Explants of leaves, basal plates, petals, anthers and ovariesof young growing corms of Crocus chrysanthus var. E. P. Bowleswere cultured on MS basal media with 20 different combinationsof either kinetin and NAA or BAP and 2, 4-D in the dark. Nomajor change was observed except on ovary explants. The ovaryexplants produced callus at 5.0 mg 1–1 and 10 mg–1BAP and subsequently stigma-like structures formed on the surfaceof the callus. Transfer to light resulted in the stigma-likestructures developing a yellow pigmentation whereupon they cameto resemble the naturally-grown stigmas. Corm formation andshoot regeneration was obtained from the callus when the ovaryexplants were cultured on media containing 5.0 and 10 mg I–1BAP with 0.5 mg 1–1 2, 4-D. Increasing the level of 2,4-D markedly reduced the number of shoots produced per explant. Key words: Crocus chrysanthus, callus, ovary explants  相似文献   

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