Effect of Phosphorus Deficiency on Phosphatase Activity of Cell Walls from Roots of Subterranean Clover

Clover (Trifolium subterraneum L. cv. Mt. Barker) was grownin solution culture with adequate (+P) or no phosphate (–P).Cell walls were extracted from roots in such a way that theywere uncontaminated by other cellular materials. Phosphataseactivity was assayed using p-nitro-phenylphosphate (NPP). Phosphatasebound to cell walls had a pH optimum between 5.0 and 6.0, irrespectiveof the P supply to the plants. Activity of phosphatase boundto cell walls increased with electrolyte concentration of theassay medium at pH 6.5 but not at pH 5.5. This increase in activitywas probably due to a higher degree of ionization of the cellwall at pH 6.5 than at pH 5.5, and to effects of high ionicstrength in decreasing the mutual repulsion of negatively chargedNPP from negative charges on the cell walls. Cell wall-boundphosphatase did not exhibit Michaelis-Menten kinetics: the concentrationof NPP at which activity was half the maximum rate (S_0.5) was0.7 mM for cell walls extracted from roots of both +P and –Pplants. Up to 30% of the phosphatase activity bound to cellwalls could be removed using buffer solutions of high pH andhigh ionic strength which contained Triton X100. Both soluble and cell wall-bound phosphatase(s) of roots increasedin activity with P deficiency. The phosphatase activity of cellwalls increased 1.5 fold as the P concentration in the rootsfell from 0.4–0.2% dry weight. Experiments with sterileroots of clover showed that increases in cell wall-bound phosphataseactivity associated with P deficiency were not due to microbialcontamination. It is argued that phosphatase(s) in cell wallsof roots could make a substantial contribution to the P nutritionof clover in soils deficient in inorganic phosphate by hydrolysingorganic phosphate compounds in the soil. Key words: Phosphatase, Clover, Roots, Phosphorus deficiency, Cell walls     

Effects of external NaCl on the growth of Atriplex amnicola and the ion relations and carbohydrate status of the leaves

Abstract Atriplex amnicola, was grown in nutrient solution cultures with concentrations of NaCl up to 750 mol m^?3. The growth optimum was at 25–50 mol m^?3 NaCl and growth was 10–15% of that value at 750 mol m^?3 NaCl. Sodium chloride at 200 mol m^?3 and higher reduced the rate of leaf extension and increased the time taken for a leaf to reach its maximal length. Concentrations of Na⁺, K⁺ and Mg²⁺ in leaves of different ages were investigated for plants grown at 25, 200 and 400 mol m^?3 NaCl. Although leaves of plants grown at 200 and 400 mol m^?3 NaCl had high Na⁺ concentrations at young developmental stages, much of this Na⁺ was located in the salt bladders. Leaves excluding bladders had low Na⁺ concentrations when young, but very high in Na⁺ when old. In contrast to Na⁺, K⁺ concentrations were similar in bladders and leaves excluding bladders. Concentrations of K⁺ were higher in the rapidly expanding than in the old leaves. At 400 mol m^?3 NaCl, the K⁺:Na⁺ ratios of the leaves excluding bladders were 0.4–0.6 and 0.1 for rapidly expanding and oldest leaves, respectively. The Na⁺ content in moles per leaf, excluding bladders, increased linearly with the age of the leaves; concurrent increases in succulence were closely correlated with the Na ⁺ concentration in the leaves excluding the bladders. Soluble sugars and starch in leaves, stems and buds were determined at dusk and dawn. There was a pronounced diurnal fluctation in concentrations of carbohydrates. During the night, most plant parts showed large decreases in starch and sugar. Concentrations of carbohydrates in most plant organs were similar for plants grown at 25 and 400 mol m^?3 NaCl. One notable exception was buds at dusk, where sugar and starch concentrations were 30–35% less in plants grown at 400 mol m^?3 NaCl than in plants grown at 25 mol m^?3 NaCl. The data indicate that the growth of A. amnicola at 400 mol m^?3 NaCl is not limited by the availability of photosynthate in the plant as a whole. However, there could have been a growth limitation due to inadequate organic solutes for osmotic regulation.     

Turgor Pressure, Volumetric Elastic Modulus, Osmotic Volume and Ultrastructure of Chlorella emersonii Grown at High and Low External NaCl

Chlorella emersonii (211/11n) was grown at external NaCl concentrationsranging between 1.0 and 335 mM (0.08–1.64 MPa). Previousstudies showed that there was no significant change in the internalconcentrations of Na⁺ or Cl^– over this range, the concentrationsremaining below 35 mM. Relative growth rates of C. emersoniiwere 30–45% lower in 335 mM NaCl than in 1.0 mM NaCl.Turgor pressure varied with the osmotic pressure of the growthmedium. Plots of cell volume versus (external osmotic pressure)^–1indicated that cells grown in 1.0 mM NaCl (0.08 MPa) had turgorpressures ranging from 0.5 to 0.8 MPa, while cells in 335 mMNaCl (1.64 MPa) had turgor pressures of 0.0–0.14 MPa.Estimates of turgor pressure derived from the osmotic pressureof cell sap had a mean value of 0.6 MPa for cells in 1.0 mMNaCl, and 0.3 MPa for cells in 335 mM NaCl. The volumetric elasticmodulus () depended on the osmotic pressure of the growth medium: was 8.5 ± 1.7 MPa for cells grown in 1.0 mM NaCl, and0.9 ± 0.6 for cells in 335 mM NaCl. was measured bychanging turgor pressures over the range 0.0–0.5 MPa,and was found to be independent of turgor. Electron micrographsshowed that the walls of cells grown in 335 mM NaCl were 70%thicker than those grown in 1.0 mM NaCl. Other changes in cellularstructure were small, however, the area occupied by vacuolesincreased from 7% in cells grown in 1.0 mM NaCl to 14% in cellsin 335 mM. The percent osmotic volume of cells grown in 1.0–335mM NaCl (61 ± 17%, v/v) was similar to the percent watercontent (59 ± 13%, w/w). Key words: Chlorella emersonii, Sodium chloride, Osmotic volume, Turgor, Volumetric-elastic-modulus     

Response of Wheat Seedlings to Low O2 Concentrations in Nutrient Solution: II. K +/Na+ SELECTIVITY OF ROOT TISSUES9

This paper reports the effects of low O₂ concentration (0–01,0–055, and 0.115mol m^–3) in nutrient solutions onK⁺/Na⁺ selectivity of growing and mature root tissues of 6-to 8-d-old, intact, wheat (Triticum aestivum cv. Gamenya) seedlings. Increases in anaerobic catabolism and decreases in O₂ uptake,K⁺ uptake and K⁺/Na⁺ selectivity were all more pronounced and/oroccurred at higher external O₂ concentrations in the apex (0–2mm) than in the expanding tissues (2–4 mm); these growingtissues were, in turn, more affected than the expanded tissuesof the roots (4–12 mm). Selectivity for K⁺ over Na⁺ in roots and shoots was particularlysensitive to O₂ deficiency. For example, in apical tissues (0–2mm) K ⁺ /Na⁺ selectivity was already reduced at 0.115 mol m^–3O₂, yet at this O₂ concentration there was no effect on eithergrowth or (K⁺/Na⁺) uptake. Upon transfer from 0.01 to 0.26 mol m^–3 O₂, a detailedstudy of the 12 mm root tips showed that 70% of these tips regainedhigh (K⁺ + Na⁺) concentrations and K⁺/^Na+ ratios. In contrast,there was no recovery in the remaining 30% of the 12 mm roottips. Net K⁺ transport to the shoots during the period afterre-aeration was negative for the population as a whole. Theseverity of these effects supports the view that the root tipsand the stele were more susceptible to O2 deficiency than wasthe cortex of the fully-developed root tissues. Key words: Hypoxia, K⁺/Na⁺ selectivity, expanded and expanding tissues     

Effects of Chloride Salts at High Concentrations on Glycolysis in vitro

KC1, at 200 mM, decreased rates of glycolysis, in vitro, inthe steady state and also increased the time required to reachthe steady state of CO₂ and ethanol formation after glucoseaddition. KCl at 200 mM increased concentrations of fructose-1,6-diphosphateand decreased concentrations of pyruvate, at all times between3 and 120 min after glucose addition. At early times after glucoseaddition, 200 mM KCl decreased concentrations of triose phosphatesand 3-phos-phoglycerate. However, at later times, concentrationsof these two intermediates became higher at 200 mM KC1 thanat low Cl^–; with triose phosphates this occurred for thefirst time at 20 min and with 3-phosphoglyeerate at 60 min afterglucose addition. These and other experiments, including oneusing the crossover theorem of Chance, strongly suggested thatthe increased concentrations of fructose- 1,6-diphosphate andtriose phosphates alleviated severe KCl inhibitions of enzymescatalysing reactions in the conversion of fructose-l,6-diphosphateto 1,3-diphosphoglycerate. Similar phenomena occurred for otherparts of the pathway. Thus, the glycolytic sequence respondedto high KCl and NaCl concentrations in a manner predicted fromearlier published experiments with single enzymes, in whichKCl and NaCl inhibitions were much smaller at high than at lowsubstrate concentrations. For the steady state, addition of purified enzymes showed thathigh KCl reduced glycolysis, at least partly by persistent inhibitionsof glyceraldehyde-3-phosphate dehydrogenase and/or aldolase. Overall, the data suggest that high KCl and NaCl concentrationshave two main effects: (1) a transient reduction in rate ofglycolysis—this effect disappears when substrates of certainenzymes have increased above the initial low levels; and (2)inhibitions which persist despite high intermediate concentrations,i.e. inhibitions which occur even in the ‘steady state’.The relevance of these in vitro data to in vivo responses athigh salt concentrations is briefly discussed.     

Effects of accumulation of 3-O-methylglucose on levels of endogenous osmotic solutes in Chlorella emersonii

Abstract. Regulation of the concentration of osmotic solutes was studied in Chlorella emersonii grown at external osmotic pressures (II) ranging between 0.08 and 1.64MPa. NaCl was used as osmoticum. The total solute content of the cells was manipulated by applying 2 mol m⁻³ 3- O -methylglucose (MG), which was not metabolized, and accumulated at concentrations ranging between 60 and 230 mol m⁻³ within 4 h after its addition to the medium. Methylglucose uptake resulted in decreases in concentrations of proline and sucrose, the two solutes mainly responsible for osmotic adaptation of C. emersonii to high external II. The responses were consistent with the hypothesis that proline and sucrose concentrations are controlled by a system of osmotic regulation, with turgor and/or volume as a primary signal. Short-term experiments showed that even very small increases in turgor and/or volume, due to accumulation of methylglucose, resulted in large decreases in proline and sucrose. Over the first 30-60 min the total solute concentration in the cells increased by at most 15 osmol m⁻³ which would represent an increase in turgor pressure of at most 0.04 M Pa. Yet, the decreases in proline and sucrose were as fast as those in cells exposed to a sudden decrease of 0.25 MPa in external II, when the turgor pressure would have increased by at least 0.15 MPa. High concentrations of methylglucose in cells grown at high II did not affect the rapid synthesis of proline and sucrose which started when the cells were transferred to yet higher II. Thus, methylglucose had no direct effects on proline and sucrose metabolism, and it has been assumed that it acted solely as an inert osmotic solute within the cell.     

Effects of a Range of O2 Concentrations on Porosity of Barley Roots and on their Sugar and Protein Concentrations

Barley was grown at a range of oxygen concentrations (0.5–9mg l^–1), in nutrient solutions. Growth of both shootsand seminal roots was restricted by O₂ concentrations lowerthan 2–3 mg l^–1) but nodal root growth was not. Root porosities were increased even at those O₂ concentrationswhich did not restrict growth, and were inversely proportionalto the protein levels of the roots. Sugar concentrations increasedappreciably only at those O₂ concentrations which also restrictedgrowth. Hordeum vulgare L., barley, root porosity, sugar, protein, oxygen concentration     

Injury to Rice Plants Caused by Complete Submergence; A Contribution by Ethylerie (Ethene)

Complete submergence of rice plants (Oryza sativa L. cv. ‘IR42’)in dilute nutrient solution for 3–6 d almost stopped theaccumulation of dry matter, depressed soluble carbohydrate concentrationby over 75% and promoted chlorosis in fully expanded leaves.Increase in fresh weight by the shoots was not impaired. Extensionby the youngest visible leaf was stimulated. Extension by thenext leaf to appear was retarded by submergence. These growthresponses to submergence were associated with a 1-5-fold increasein the partial pressure of endogenous ethylene (ethene). Applying ethylene (0.3–0.35 Pa) in the gas-phase to non-submergedplants reproduced some, but not all, of these effects of submergence.Thus, greater leaf extension and chlorosis of submerged plantscould be attributable to accumulated ethylene but neither theslow relative growth rate nor the decreased extension of leavesemerging after the start of submergence could be so attributed. Two cultivars (‘FR13A’ and ‘Kurkaruppan’)already known to tolerate submergence, differed little fromsubmergence-intolerant ‘IR42’ in their relativegrowth rate and soluble carbohydrate concentration during submergence.However, their underwater leaf extension was less than in ‘IR42’and chlorosis was much less prevalent, especially in ‘FR13A’.Similarly, ethylene supplied to non-submerged plants was a lesseffective promotor of leaf extension and chlorosis in the twosubmergence tolerant cultivars. Application of 1.0 kPa carbondioxide in the gas-phase prevented the chlorosis response toethylene. The results indicate that accumulated ethylene is a likely causeof fast leaf extension and chlorosis in submergence intolerantforms of rice, particularly when amounts of dissolved carbondioxide are minimal. Key words: Oryza sativa L., aeration, ethylene (ethene), stress-tolerance