Osmotic Stress in Coleoptiles and Primary Leaves of Wheat: I. EFFECTS ON SIZE, WEIGHT, TOTAL PROTEIN, DNA, AND PHOSPHORUS

Seedlings of wheat (Triticum durum, cv. Balcarceño-INTA)were water-stressed in darkness with 20% polyethylene glycol(PEG) 6000 or 0.3 M mannitol added to the root medium. At differenttimes and up to a total of 36 h of treatment the coleoptileand primary leaves were cut and analysed. The height and freshweight of shoots were lower in treated plants than in controlplants. Dry weight was not significantly different between controland water-stressed plants. Total protein concentration decreasedsignificantly (P < 0.01) after 36 h of PEG 6000 treatment.Total DNA concentration decreased in controls but not significantly(P < 0.025) in treated seedlings. This result was interpretedas indicating that cell elongation prevailed over cell divisionin controls and that cell enlargement was affected in stressedplants. Total phosphorus concentration fell in control and treatedseedlings. However, phosphorus specific radioactivity increasedby 116% in control plants, 93% in mannitol-treated plants, and22% in PEG 6000-treated seedlings. These data suggest that anearly metabolic effect of water stress may be on phosphorusturnover in shoots.     

Mild phosphorus stress in barley and a related low-phosphorus-adapted barleygrass: Phosphorus fractions and phosphate absorption in relation to growth

Barleygrass ( Hordeum leporinum ) from Australian low-P (phosphorus) soils and commercial barley ( H. vulgare ) with high fertilizer requirements were grown in solution culture at 3 levels of P supply. The high-P-adapted barley produced more biomass at all levels of P supply and was more responsive to added P in terms of rate of tillering, rate of leaf production, final leaf size, and therefore total shoot weight compared to barleygrass. In both species root: shoot ratio decreased in response to improved tissue P status, even at P levels where total biomass did not respond to P supply. Removal of endosperm reserves of barley reduced total biomass to a greater extent than it altered phosphate absorption rate, thus increasing tissue P status and making plants less responsive to added P. Similarly, barleygrass had a slower growth rate but a comparable P absorption rate to that of barley. Thus barleygrass also accumulated tissue P and was unresponsive to added P. All phosphorus chemical fractions increased in response to improved tissue P status, but to differing extents (inorganic-P > nucleic acid-P > lipid-P > ester-P), suggesting that all P fractions (particularly inorganic P) serve, in part, a storage function. Both barleygrass and barley without endosperm had higher concentrations of all P fractions (particularly inorganic P) than did unaltered barley, but this was due entirely to their higher P status (due to slow growth) rather than to any major difference in P metabolism between species. We conclude that slow growth is more important than interspecific differences in P metabolism, P absorption, or efficiency of P utilization in explaining the success of barleygrass and other low-P-adapted species on infertile soils.     

Turgor, Growth and Rheological Gradients of Wheat Roots Following Osmotic Stress

The growth rate of hydroponically grown wheat roots was reducedby mannitol solutions of various osmotic pressures. For example,following 24 h exposure to 0·96 MPa mannitol root elongationwas reduced from 1· mm h^–1 to 0·1 mm h^–1 Mature cell length was reduced from 290 µm in unstressedroots to 100 µm in 0·96 MPa mannitol. This indicatesa reduction in cell production rate from about 4 per h in theunstressed roots to 1 per h in the highest stress treatment. The growing zone extended over the apical 4·5 mm in unstressedroots but became shorter as growth ceased in the proximal regionsat higher levels of osmotic stress. The turgor pressure along the apical 5·0 mm of unstressedroots was between 0·5 and 0·6 MPa but declinedto 0·41 MPa over the next 50 mm. Following 24 h in 0·48(200 mol m^–3) or 0·72 MPa (300 mol m^–) mannitol,turgor along the apical 50 mm was indistinguishable from thatof unstressed roots but turgor declined more steeply in theregion 5·10 mm from the tip. At the highest level ofstress (0·96 MPa or 400 mol m^–3 mannitol) turgordeclined steeply within the apical 20 mm. Key words: Growth, turgor pressure, wall rheology, osmotic stress, osmotic adjustment     

NONDIALYZABLE SULFATED SIALOGLYCOPEPTIDE FRACTIONS DERIVED FROM BOVINE HEIFER BRAIN GLYCOPROTEINS. ISOLATION,CHARACTERIZATTON AND CARBOHYDRATE-PEPTIDE LINKAGE STUDIES1

The nondialyzable sialoglycopeptides dcrived from defatted, protease digested whole boyine heifer brain tissue were isolated and characterized. The partially purified bovine heifer brain tissue were isolated and characterized. The partially purified sialoglycopeptide mixtures contained 7-8% ester sulfate. The amount of nonsulfated material present was of the order of 15%. The total mixture was fractionated by anion exchange chromatography on AG1-X2(CI^-) using a sodium chloride gradient to yield twelve fractions all of which contained ester sulfate to varying degrees (5.6-21.9%). The molecular size of the fractions ranged from molecular weights of 10,400-12,500. The molar sulfate/galactose plus glucosamine ratios ranged from 0.3 to 1.1. Carbohydrate peptide linkage studies of the fractions by treatment with 0.2 M-NaOH-O.3 M-NaBH₄ led to the destruction of a portion of the threonine, serine and galactosamine and the appearance in acid hydrolysates of α-amino-n-butyric acid and galactosaminitol with an increase in alanine. Partial acid hydrolysis of the most abundant sialoglycopeptide liberated detectable amounts of 2-acetarnido-1-(L-4-aspartamido-)-1,2-dideoxy-β-D- glucose. These results indicated that the GalNac at the reducing end of the alkali labile carbohydrate prosthetic groups were linked O-glycosidically to thc hydroxyl groups of threonine and serine and the alkali-stable region of the principal sialoglycopeptide involves N-acetylglycosaminyl-asparagine linkages. Mild acid hydrolysis of the alkali-stable portion of the sulfated fractions indicated that the ester sulfate was confined to the alkali-stable region and exists as galactose 6-O-sulfate and N-acetylglucosa- mine 6-O-sulfate, structural fcatures similar to those present in rat brain glycopcptides (Markgolis & Margolis , 1970; Margolis et al.; 1972). Fucose and ester sulfate were confined to the alkali-stable region while mannose was located in both the alkali-labilc and alkali-stable regions of the sialoglyco-peptide fractions.     

Ascorbate enhancement of H1 histamine receptor sensitivity coincides with ascorbate oxidation inhibition by histamine receptors

Ascorbate has previously been shown to enhance both ₁- and ₂-adrenergic activity. This activity is mediated by ascorbate binding to the extracellular domain of the adrenergic receptor, which also decreases the oxidation rate of ascorbate. H1 histamine receptors have extracellular agonist or ascorbate binding sites with strong similarities to _1- and ₂-adrenergic receptors. Physiological concentrations of ascorbate (50 µM) significantly enhanced histamine contractions of rabbit aorta on the lower half of the histamine dose-response curve, increasing contractions of 0.1, 0.2, and 0.3 µM histamine by two- to threefold. Increases in ascorbate concentration significantly enhanced 0.2 µM histamine (5–500 µM ascorbate) and 0.3 µM histamine (15–500 µM ascorbate) in a dose-dependent manner. Histamine does not measurably oxidize over 20 h in oxygenated PSS at 37°C. Thus the ascorbate enhancement is independent of ascorbate's antioxidant effects. Ascorbate in solution oxidizes rapidly. Transfected histamine receptor membrane suspension with protein concentration at >3.1 µg/ml (56 nM maximum histamine receptor) decreases the oxidation rate of 392 µM ascorbate, and virtually no ascorbate oxidation occurs at >0.0004 mol histamine receptor/mol ascorbate. Histamine receptor membrane had an initial ascorbate oxidation inhibition rate of 0.094 min·µg protein^–1·ml^–1, compared with rates for transfected ANG II membrane (0.055 min·µg protein^–1·ml^–1), untransfected membrane (0.052 min·µg protein^–1·ml^–1), creatine kinase (0.0082 min·µg protein^–1·ml^–1), keyhole limpet hemocyanin (0.00092 min·µg protein^–1·ml^–1), and osmotically lysed aortic rings (0.00057 min·µg wet weight^–1·ml^–1). Ascorbate enhancement of seven-transmembrane-spanning membrane receptor activity occurs in both adrenergic and histaminergic receptors. These receptors may play a significant role in maintaining extracellular ascorbate in a reduced state. molecular complementarity; vitamin C; seven-transmembrane-spanning membrane receptors     

Changes in sediment phosphate composition of seasonal ponds during filling

Most ponds from the Doñana National Park are shallow temporary freshwater bodies on eolian sands. The total phosphate concentration and the fractional P-composition of the sediments from two small ponds were studied before and inmediately after they were filled (autumn, 1996). Total phosphorus concentration was measured in 2 different size fractions: 2–0.1 mm (coarse) and <0.1 mm (fine). In both ponds, the total phosphorus concentration of the fine sediment increased by 20% at the beginning of the filling period, whereas 5 weeks later it did not increase further. The percentage of organic matter of the fine sediment was relatively high (between 13–18%) and did not change significantly during filling. The concentration of sediment total iron and Fe(OOH) increased significantly in both ponds during filling. The sequential P fractionation of the fine sediment included, besides the determination of two inorganic fractions (Ca-bound P and Fe-bound P), an organic-P fraction extracted with acid, another extracted with alkali, two organic-P fractios from the Ca-EDTA/dithionite and the Na₂-EDTA extracts which contained high concentrations of humic substances. The pond sediments were rich in organic P compounds as the sum of all organic-P fractions ranged between 267 and 320 g g^-1 (68% and 79% of the sum of all fractions). Significant changes (P<0.01) in the fractional P-composition of sediments were found after filling. Acid soluble organic phosphate increased about 30% in both ponds. Iron-bound phosphate increased significantly (about 40%) only in the pond where a higher concentration of Fe(OOH) was measured after filling. An adsorption experiment was carried out for each sediment to simulate P input during filling. Both the iron-bound phosphate and Ca-bound P increased significantly (P<0.01) suggesting that these two fractions were involved in the P-adsorption during the laboratory experiment.     

Cryopreservation of Shoot Tips of Tetraploid Potato (Solanum tuberosumL.) Clones by Vitrification

In vitro-grown shoot tips of five tetraploid potato (SolanumtuberosumL.) clones were cryopreserved by vitrification. Excisedshoot tips (0.5–0.7 mm) were pre-cultured on filter paperdiscs over half strength liquid Murashige and Skoog (MS) mediumsupplemented with 8.7 µMGA₃and different combinationsof sucrose (0.3, 0.5 and 0.7M) plus mannitol (0, 0.2 and 0.4M)for 2 d under a 16 h photoperiod at 24 °C. The pre-culturedshoot tips were either successively loaded with 20 and 60% PVS2 solutions or directly exposed to concentrated vitrificationsolution before physical vitrification during liquid nitrogentreatment. The vitrified shoot tips were warmed rapidly andtreated with dilution mixture (MS+1.2Msucrose) for 30 min beforeplating on regrowth medium. Addition of mannitol to the pre-culturemedium improved survival of vitrified shoot tips. Direct dehydrationof pre-cultured shoot tips with concentrated PVS 2 was detrimentalto survival of vitrified shoot tips. Shoot tips pre-culturedon medium containing 0.3Msucrose plus 0.2Mmannitol, and loadedwith 20% PVS 2 for 30 min followed by 15 min incubation in 60%PVS 2 and 5 min incubation in 100% PVS 2 at 0 °C resultedin up to 54% survival after vitrification. About 50% of vitrifiedand warmed shoot tips formed shoots directly. Post-thaw culturingof vitrified shoot tips on medium containing an elevated levelof sucrose (0.2M) under diffuse light for the first week enhancedthe survival rate. Continuous culturing of vitrified shoot tipson high-sucrose medium induced multiple shoot formation.Copyright1998 Annals of Botany Company Solanum tuberosumL., potato, cryopreservation, germplasm conservation,in vitroconservation, meristems, shoot tips, tissue culture, vitrification.     

Effect of Water Stress on Gas Exchange in Fronds of the Ostrich Fern (Matteuccia struthiopteris (L.) Todaro)

Experiments were conducted in a gas exchange system to examinethe effect of a water stress, induced by –200 kPa polyethyleneglycol (PEG), on carbon dioxide and water vapour flux, fronddiffusive resistance, intercellular carbon dioxide concentration,carbon dioxide residual resistance and frond water potentialin the ostrich fern (Matteuccia struthiopteris (L.) Todaro).Measurements were taken 1 d after the application of PEG. Themeasurements were made on young fronds (8 d old) and maturefronds (20–24 d old) at PPFD's (Photosynthetic PhotonFlux Density) from 0–1400 µmol m^–22 s^–1.Water stress decreased the net photosynthesis rate in maturefronds at PPFD's of 210 µmol m^–2 s^–1 or greaterand increased the net photosynthesis rate below 210 µmolm^–2 s^–1 in young fronds. The increase in net photosynthesisin stressed young fronds was associated with a significant reductionin the dark respiration rate. Water stress and decreasing PPFD'sincreased frond diffusive resistance. Carbon dioxide concentrationin the intercellular spaces decreased with increasing frondage and PPFD's up to 200 µmol m^–2 s^–1. Theresidual resistance to carbon dioxide flux was not significantlyaffected by either frond age or water stress. Frond water potentialwas significantly lower in mature fronds than in young fronds. Key words: Matteuccia struthiopteris, Water relations, Photosynthesis, Dark respiration     

Polyethylene Glycol Induced Fusion of Selected Pairs of Single Protoplasts

A new method for polyethylene glycol (PEG) -induced fusion between single pairs of selected protoplasts was developed. The protoplasts were prepared from tobacco leaves. Under an inverted microscope two defined protoplasts were selected with a hand-made micropipette and transferred into a droplet of fusion solution containing 25 % PEG (M. W. 6000), 0. 1 mol/L mannitol and 0. 01 mol/L CaCl2 · 2H2O (pH 5.6). Slightly moving the pipette caused the protoplasts to contact and adhere to each other, the fusion pairs were then transferred to a solution containing 10% PEG, 0.35 mol/L sucrose and 0. 01 mol/L CaCl2 · 2H2O (pH 5.6) for approximately 10 min, followed by subsequent washing with a solution containing 0.45 mol/L sucrose and 0.04 mol/L CaC12 · 2H20 (pH 7—9). Compared with conventional fusion methods adopted to protoplast population, the present method can avoid either blind fusion of protoplasts belonging to one partner and fusion among multiple protoplasts, or the presence of unfused protoplasts, thus ensure the fusion to be precisely at the level of a selected pair of single protoplasts. Moreover, it is simple and convenient enough to show its potentiality for wide application in somatic hybridization and particularly in the case of small quantity of parental protoplasts such as in vitro intergametic fusion studies.     

Relations between MAPK phosphorylation and ABA level in <Emphasis Type="Italic">Malus sieversii</Emphasis> under water stress

A correlation between protein kinase phosphorylation and ABA level was studied in Malus sieversii (Ledeb.) Roem. seedlings under water stress. The seedlings were treated with PEG 6000 for imitation of water stress, and the MAPK activity and ABA content in each treatment were then determined. We demonstrated that the increase in the activities of the total protein kinase (TPK) and mitogen-activated protein kinase (MAPK) after treatment with 20% PEG 6000 appeared to result in a high level of ABA. MAPK activity accounted for 76.8% of TPK activity. The activity peaks of TPK and MAPK preceded the highest level of ABA accumulation. It is interesting that the ABA level in roots and leaves of seedlings pretreated with 2 × 10⁻² mM exogenous ABA for 20 min following treatment by 20% PEG 6000 was much higher than that of seedlings treated with exogenous ABA only. We analyzed the influence of MAPK inhibitor ITU (5-iodotubercidin) on ABA accumulation in the seedlings of M. sieversii under water stress and showed that 1 μM ITU significantly decreased the ABA level induced by a water loss. However, the phosphoesterase inhibitor PAO (phenylarisine oxide) enhanced ABA accumulation, indicating that the phosphorylated MAPK was correlated to ABA synthesis. Together, these results suggest that MAPK phosphorylation played an important role in ABA accumulation under water stress, and MAPK might mediate the signal transduction of ABA synthesis.     

Measurement of Yield Threshold and Cell Wal Extensibility of Intact Wheat Roots under Different Ionic, Osmotic and Temperature Treatments

Yield stress threshold (Y) and volumetric extensibility () arethe rheological properties that appear to control root growth.In this study they were measured in wheat roots by means ofparallel measurement of the growth rate (r) of intact wheatroots and of the turgor pressures (P) of individual cells withinthe expansion zone. Growth and turgor pressure were manipulatedby immersion in graded osmoticum (mannitol) solutions. Turgorwas measured with a pressure probe and growth rate by visualobservation. The influence of various growth conditions on Yand was investigated; (a) At 27 °C.In 0.5 mol m^–3 CaCl₂ r, P, Y and were20.7±4.6 µm min^–1, 0.77±0.05 MPa,0.07±0.03 MPa and 26±1.9 µm min^–1MPa^–1 (expressed as increase in length), respectively.Following 24 h growth in 10 mol m^–3 KC1 these parametersbecame 12.3±3.5 µm min^–1, 0.72±0.04MPa, 0.13±0.01 MPa and 21±0.7 µm min^–1MPa^–1. After 24 h osmotic adjustment in 150 mol m^–3mannitol/0.5 mol m^–3 CaCl₂ r= 19.6±4.2 µmmin^–1, P = 0.68±0.05 MPa and Y and were 0.07±0.04MPa and 30±0.2 µm min^–1 MPa^–01, respectively.After 24 h growth in 350 mol m^–3 mannitol/0.5 mol m^–3CaCl₂ r= 13.3±4.1 µm min^–1, P= 0.58±0.07MPa, Y=0.12±0.01 MPa and ø 32±0.2 tim min^–1MPa^–1. During osmotic adjustment in 200 mol m^–3mannitol/0.5 mol m^–3 CaCl₂, with or without KCl, the recoveryof growth rate corresponded to turgor pressure recovery (t1/2approximately 3 h). (b) At 15 °C. Lowered temperature dramatically influencedthe growth parameters which became r= 8.3±2.8 um min^–1,P=0.78 MPa, r=<0.2 MPa and =15±0.1 µm min^–1MPa^–1. Therefore, Y and are influenced by 10 mol m^–3 K⁺ ionsand low temperature. In each case the effective pressure forgrowth (P-Y) was large indicating that small fluctuations ofsoil water potential will not stop root elongation. Key words: Yield threshold, cell wall extensibility, wheat root growth, temperature, turgor pressur     

Studies of pesticides effects on Chlorella metabolism III. Effect of isopropyl 3-chlorocarbanilate (chlorpropham) on cell cycle and biosynthesis

Isopropyl 3-chlorocarbanilate (chlorpropham) inhibited Chlorellagrowth by 50% at 1.3 µM under non-photosynthetic conditions.Average DNA content per cell was increased 2.5-fold by chlorprophamtreatment at 4.7 µM. Oxygen uptake was not significantlyaffected at 47 µM. Protein synthesis was more sensitiveto chlorpropham than the other biosynthetic processes tested,but it was not inhibited at 1.3 µM. In cell cycle studiesperformed under photosynthetic conditions, 50% inhibition ofgrowth occurred at 4 µM. At 14 µM, growth in termsof cell number was completely suppressed while inhibition ofgrowth in terms of average cell volume was partial, resultingin comparatively larger cell volume. This was accompanied by3.0-fold increase in average DNA content per cell. (Received March 8, 1976; )     

Effect of Plasticizer on Release Kinetics of Diclofenac Sodium Pellets Coated with Eudragit RS 30 D

The present study was designed to investigate the effect of two plasticizers, i.e., triethyl citrate (TEC) and polyethylene glycol 6000 (PEG 6000) on the in vitro release kinetics of diclofenac sodium from sustained-release pellets. Ammonio methacrylate copolymer type B (Eudragit RS 30 D) is used as the release-retarding polymer. Both plasticizers were used at 10% and 15% (w/w) of Eudragit RS 30 D. Pellets were prepared by powder layering technology and coated with Eudragit RS 30 D by air suspension technique. Thermal properties of drug and drug-loaded beads were studied using differential scanning calorimeter (DSC). DSC thermogram represented the identity of raw materials and exhibited no interaction or complexation between the active and excipients used in the pelletization process. Dissolution study was performed by using USP apparatus 1. No significant difference was observed among the physical properties of the coated pellets of different batches. When dissolution was performed as pure drug, about 8.22% and 90% drug was dissolved at 2 h in 0.1 N HCl and at 30 min in buffer (pH 6.8), respectively. From all formulations, the release of drug in acid media was very negligible (maximum 1.8 ± 0.08% at 2 h) but in buffer only 12% and 30% drug was released at 10 h from coated pellets containing TEC and PEG 6000, respectively, indicating that Eudragit RS 30 D significantly retards the drug release rate and that drug release was varied according to the type and amount of plasticizers used. The amount of TEC in coating formulation significantly effected drug release (p < 0.001), but the effect of PEG 6000 was not significant. Formulations containing PEG 6000 released more drug (98.35 ± 2.35%) than TEC (68.01 ± 1.04%) after 24 h. Different kinetic models like zero order, first order, and Higuchi were used for fitting drug release pattern. Zero order model fitted best for diclofenac release in all formulations. Drug release mechanism was derived with Korsmeyer equation.     

Implications of zooplankton stoichiometry on distribution of N and P among planktonic size fractions

Pooled samples from the upper 20 m at five stations in the Oslofjord(Norway) were size fractionated and analysed for particulatedry weight, carbon, nitrogen and phosphorus. The nano fraction(0.7–20 µm) dominated in biomass throughout thesampling period. The C:N ratios of the fractions did not differmuch from each other. The C:P ratios of the nano- and microfraction(20–200 µm) were considerably higher than the ratioof the mesofraction (200–2000 µm) throughout thesampling period. High C:P ratios and low phosphate concentrationsabove the pycnocline suggest that the system was P-limited.The stoichiometry of mesozooplankton was more constant thanthe stoichiometry of the other fractions, and the zooplanktonconstituted consistently a higher percentage of the phosphoruspool than of the carbon pool. This suggests that the mesozooplanktoncan act as a sink of nutrients due to its invariable stoichiometry.     

Modulation of cardiac PIP2 by cardioactive hormones and other physiologically relevant interventions

Phosphatidylinositol4,5-bisphosphate (PIP₂) affects profoundly several cardiacion channels and transporters, and studies ofPIP₂-sensitive currents in excised patches suggest thatPIP₂ can be synthesized and broken down within 30 s.To test when, and if, total phosphatidylinositol 4-phosphate (PIP) andPIP₂ levels actually change in intact heart, we used a new,nonradioactive HPLC method to quantify anionic phospholipids. Total PIPand PIP₂ levels (10-30 µmol/kg wet weight) do notchange, or even increase, with activation ofG_q/phospholipase C (PLC)-dependent pathways by carbachol(50 µM), phenylephrine (50 µM), and endothelin-1 (0.3 µM).Adenosine (0.2 mM) and phorbol 12-myristate 13-acetate (1µM) bothcause 30% reduction of PIP₂ in ventricles, suggesting thatdiacylglycerol (DAG)-dependent mechanisms negatively regulate cardiacPIP₂. PIP₂, but not PIP, increases reversiblyby 30% during electrical stimulation (2 Hz for 5 min) in guinea pigleft atria; the increase is blocked by nickel (2 mM). Both PIP andPIP₂ increase within 3 min in hypertonic solutions, roughlyin proportion to osmolarity, and similar effects occur in multiple celllines. Inhibitors of several volume-sensitive signaling mechanisms do not affect these responses, suggesting that PIP₂ metabolismmight be sensitive to membrane tension, per se.

     

The Diffusion of KCI from Micro-electrodes

The diffusion of salt from the tips of 3 M KCI filled micro-electrodesis measured. The leakage of KCI in 3 min is found to be 5.70x 10^–12–1.08 x 10^–11 mol for tips of 0.2–0.6µm diameter, and 2.70 x 10^–11 mol for tips of 0.6–1.0µm diameter. The observed leakages compare with estimatesbased on the diffusion coefficient of KCI and micro-electrodegeometry. The results confirm that the use of 3 M KCI in micro-electrodesmay lead to serious contamination of impaled cells providedthe electrode tips remain unoccluded.     

A 796?bp PsPR10 gene promoter fragment increased root-specific expression of the GUS reporter gene under the abiotic stresses and signal molecules in tobacco

A 1681 bp PsPR10 promoter was isolated from Pinus strobus and a series of 5′-deletions were fused to the β-glucuronidase (GUS) reporter gene and introduced into tobacco. GUS activity in P796 (−796 to +69) construct transgenic plant roots was similar with that of P1681 and higher than those of the P513 (−513 to +69) and P323 (−323 to +69) transgenic plants. Moreover, the abiotic stresses of NaCl, PEG 6000 and mannitol, and salicylic acid (SA), abscisic acid (ABA) and jasmonic acid (JA) induced higher GUS activity in the roots of P796 transgenic tobacco. This study provides a potential inducible root-specific promoter for transgenic plants.     

The Carbon Economy of Clonal Plants of Trifolium repens L.

Fluxes of carbon between sources and sinks were quantified forclonal plants of Trifolium repens L. (cv. Blanca) in two glasshouseexperiments. Carbon sources were (a) leaves on the parent (=main)stolon apex, or (b) leaves on either young or old branches,and the major sinks of interest were the parent stolon apex,branches, and the adventitious root arising at the same parentstolon node as a young source branch. Defoliation treatmentswere applied to the parent stolon and/or branches (excludingsource branches). Carbon moved freely from the parent stolon to branches and vice-versa;these bidirectional exchanges of C provided important supplementarysources of carbohydrate for the sinks and buffered them againstthe effects of defoliation. Young branches exported more C tothe parent plant (mean=6.3µmol d^–1) than they importedfrom leaves on the parent stolon (5·2µmol d^–1)which, in turn, exceeded the amount fixed by leaves on the branchand utilized within the branch itself (2·7µmold^–1). In contrast, the C economy of old branches was largelyself-contained with, on average, 25·4µmol d^–1exported to the parent plant, 1·8µmol d^–1imported from the parent, and 63·0µmol d^–1fixed and utilized by the branch itself. Thus the growth ofyoung branches was immediately reduced by removal of parentstolon leaves, but old branches were unaffected. An estimated 42% of the C utilized by the main stolon apex originatedfrom branches, while by far the largest proportion (84%) ofthe C used for growth of young nodal roots originated from theassociated branch and not from leaves on the parent stolon towhich the root was directly attached. Key words: Trifolium repens, clonal growth, carbon economy, physiological integration, defoliation     

A Comparison Between the ATPase and Proton Pumping Activities of Plasma Membranes Isolated from the Stele and Cortex of Zea mays Roots

Plasma membrane vesicles of high purity, determined by markerenzyme assays, were obtained by phase partitioning microsomalfractions from stelar and cortical tissues of Zea mays (cv.LG11) roots. ATP hydrolytic activities in both of the plasmamembrane fractions were inhibited by vanadate, SW26 and erythrosinB, but were insensitive to nitrate. Activity in both fractionsexhibited a marked pH optimum of 6·5 and displayed typicalMichaelis-Menten kinetics. A high substrate specificity wasapparent in both the stele and cortex plasma membrane fractions,while the lower fractions, after phase partitioning, showedlower specificity for nucleotide substrates. Specific activitiesof the stele (67·8 µmol Pi mg^–1 h^–1)and cortex (78·4 µmol Pi mg^–1 h–¹)plasma membrane H⁺ -ATPases were very similar. Proton pumping activities in microsomal membrane fractions fromstele and cortex were inhibited by nitrate and insensitive tovanadate. Homogenization of stele and cortex tissue in the presenceof 250 mol m^–3 KI resulted in microsomal fractions exhibitingvanadate-sensitive, nitrate-insensitive proton pumping activity,suggesting a plasma membrane origin for this activity. SW26was also an effective inhibitor of proton pumping activity,although results indicated an interaction between SW26 and thefluorescent probes quinacrine and acridine orange. The results are discussed in relation to models for the transportof ions into the stele and are consistent with a role for theH⁺ -ATPase activity in this process. Key words: ATPase, cortex, plasma membrane, stele, Zea mays