The physiological significance of cyanide-resistant respiration in higher plants

Abstract A brief survey of the biochemistry of the alternative oxidative pathway (‘cyanide-resistant respiration’) and its occurrence in vivo is given. Several hypotheses about the physiological significance of the alternative chain are discussed. These include a role in (1) heat production, (2) fruit ripening, (3) respiration of plants that contain high levels of cyanogenic glycosides, producing HCN upon wounding, (4) oxidation of NADH that is produced by various causes in excess of that required for ATP production, (5) ion uptake, and (6) osmoregulation. In intact roots of higher plants, the activity of the alternative pathway is more active when less carbohydrate is required for assimilation of N (NH+₄ NO-₃ or N₂) and is less active in those when carbohydrates are being stored in a storage organ or when the availability of photosynthate is reduced. An increase in carbohydrate requirement for osmoregulation is also correlated with decreased alternative chain activity. It is concluded that the alternative pathway in roots plays an important role in oxidation of sugars which are not required for carbon skeletons, energy production for growth and maintenance processes, osmoregulation or storage. However, the significance of this role may vary in different tissues and physiological states.     

A systematic approach to the simulation of short-term processes in the plant-environment complex

Abstract. A conceptual framework is presented for modelling short-term processes in the plant and its environment as an integrated system. Flows of water, water vapour, heat, momentum, CO₂, soluble carbohydrate and phosphorus are all described by equations of the same general type, i.e. in terms of diffusivity-type parameters, capacities and potential gradients. A representative volume of the crop is divided horizontally into layers and vertically between crop and environment for treatment by a finite-difference method. Vertical flow occurs in the atmosphere, soil, stems and larger roots, andilateral flow between leaves and air, and between finer roots and soil. The interception of direct sunlight and the flux densities of downward and upward diffuse radiation within layers are calculated by a step-wise procedure.
The conversions of materials within the plant are treated as functions of appropriate state variables. Schemes for carbon and phosphorus provide for flow to and from the translocation system, and for photosynthesis, respiration and growth.
A model of a fully-established lucerne crop is described and the sensitivity of model performance to changes in a number of parameter values explored. Simulation runs under varying conditions indicate realistic prediction of diurnal trends.     

The Effect of a Quaternary Amine (Aliquat 336) on Growth and Photosynthesis of the Green Alga Chlorella emersonii, and the Effect on Photosynthesis in Isolated Spinach Chloroplasts

A quaternary amine, Aliquat 336, inhibits the growth of the green alga Chlorella emersonii, ¹⁴C-fixation of the alga is also inhibited. The effect and the site of action of the compound was studied by using isolated spinach chloroplasts. The carbon dioxide dependent oxygen evolution of the chloroplasts is inhibited directly upon the addition of the amine and the oxygen evolution is replaced by an oxygen uptake. By investigating some electron transport reactions in the chloroplasts we were able to show that Aliquat 336 affects the electron transport on the level of photophosphorylation. The results from the in vivo and the in vitro experiments thus show that the quaternary amine affects the photosynthetic process. Aliquat 336 is a solvent extractant used in several industrial processes for extraction of metals from aqueous solutions. Aliquat 336 could be considered a presumptive water pollutant as the compound could enter a recipient water body and thus affect photosynthesis.     

Ionic Balance of Root-Shoot Nitrate Transfer in Dwarf Bean

The Dijkshoorn-Ben Zioni model about potassium recycling in higher plants proposes a stoichiometric coupling between the upward transport of nitrate and potassium, and therefore a 1:1 ratio of these ions in the xylem sap. Phaseolus vulgaris grown in media with different NO₃: K⁺ ratios, however, only rarely showed equimolar concentrations of the two ions in the bleeding sap. Ratios between 0.2 and 11 were observed and the ratios between concentrations of the ions in the exudate and in the external solution were positively correlated. An analysis of the major inorganic constituents of the exudate revealed an ionic balance that requires negligible amounts of charges from organic molecules.     

Respiration of the Roots of Flood-Tolerant and Flood-Intolerant Senecio Species: Affinity for Oxygen and Resistance to Cyanide

The affinity of respiration for oxygen in the roots of six Senecio species studied was low compared with the affinity of cytochrome oxidase for oxygen. Half saturation values of approximately 22 μM oxygen were measured. Root respiration was to a large extent insensitive to cyanide in flood-tolerant as well as in flood-sensitive species. The evidence presented suggests that high activity of salicylhydroxamic acid (SHAM)-sensitive oxidase in Senecio roots was the basis for the low oxygen affinity and for the high cyanide-insensitivity of root respiration in the Senecio species. Methods are described to determine the in vivo activity of the SHAM-sensitive oxidase. It was estimated that it contributed 70% to the total root respiration. The presence of SHAM-sensitive oxidase activity could explain a higher efficiency of root growth respiration under a low oxygen tension if this alternate oxidase was inhibited at a low oxygen concentration in the root medium. However, the SHAM-sensitive oxidase was not specifically involved in either growth respiration or maintenance respiration. Its significance in regulation of the redox state of the cells is discussed.     

Kinetics of Integration of Xyloglucan into the Walls of Suspension-Cultured Rose Cells

To study the kinetics of synthesis, wall-binding and degradationof xyloglucan, we incubated suspension-cultured rose cells for0–5–24 h in L-[1-³H]arabinose. >95% of the [³H]arabinosewas taken up within 2 h. UDP-Pentoses were maximally labelledwithin 0–5 h and had lost most of their ³H by 2 h afterthe addition of [³H]arabinose. Therefore, the 24 h experimentresembled a pulse-chase rgime. The [³H]xyloglucan formed wasfractionated into four cellular pools [detergent-extractable(interpreted as cytoplasmic), and guanidinium thiocyanate-,06 M NaOH- and 60 M NaOH-extractable (interpreted as progressivelymore firmly wall-bound)]; soluble extracellular xyloglucan wascollected as a fifth pool. All five pools of xyloglucan hadstarted accumulating ³H at their respective maximal rates by     

Differentiation of New Metastatic Variants of B16 Melanoma Under Different Culture Conditions

The purpose of this study was to examine the differentiation of variant tumors of the B16 metastatic melanoma when tumors were grown serially under different culture conditions and transplanted into C57BL/6J black mice, lethal yellow A^y/a, albino c/c, and C⁺/c mutant mice. Morphological and biochemical markers of melanogenesis were examined in cells in culture and in the corresponding tumors. Cellular pigmentation was assessed in terms of the levels of DOPA and 5-S-CD and in terms of tyrosinase activity in the various cell lines and tumors. The observed change from high to low metastatic capacity, which was dependent on culture conditions, appeared to be unrelated to melanogenesis even though changes were observed in the biochemical melanotic phenotype. Overall, tumor cells from spontaneous pulmonary metastases appear to differentiate in ways that are unrelated to the instability of experimental metastatic capacity. The melanotic phenotype in albino c/c and C⁺/c mice was dependent on the phenotype of the parental tumors. A marked difference was observed between two pigmentation compartments, one of which was stable in the B16 control, while the other was unstable in YB16 and MB16 variant cells and in the tumors derived from them. It appears, therefore, that the metastatic capacity of B16 metastatic variants is changeable and is independent of the unstable melanogenic behavior. The production of metastases and the differentiation of tumors in the present experiments appeared to be related to the genetic background of the mice and the epigenetic metabolic environment of tumors and cells.     

Effect of Plant Growth Regulators on Nitrate Utilization by Roots of Nitrogen-Depleted Dwarf Bean

We examined the effect of pretreatments (18 h at 5 µmoldm^–3) with abscisic acid, the ethylene-releasing substance‘Ethephon’, gibberellic acid, indoleacetic acid,kinetin and zeatin on nitrate uptake and in vivo nitrate reductaseactivity (NRA) in roots of nitrogen-depleted Phaseolus vulgarisL. Nitrate uptake showed an apparent induction pattern witha steady state after about 6 h, in all treatments. The nitrateuptake rate after 6 h was unaffected or at most 30% lower aftertreatments with the plant growth regulators. Gibberellic acid, kinetin and zeatin induced substantial NRAin roots in the absence of nitrate, whereas Ethephon enhancedNRA only during nitrate nutrition. Kinetin-induced NRA (Ki-NRA)was maximal after a pretreatment at 1 µmol dm^–3,and showed a lag phase of 6–8 h. Ki-NRA was additive tonitrate-induced NRA (NO^–₃-NRA) for at least 24 h, independentof the induction sequence. After full induction, Ki-NRA approximated20% of NO-₃-NRA. Abscisic acid counteracted the developmentof Ki-NRA, but not of NO^–₃-NRA. Cycloheximide and tungstatewere equally effective to suppress the development of nitratereductase activity after supply of kinetin or NO^–₃. Our data are consistent with the operation of two independentenzyme fractions (Ki-NRA and NO^–₃-NRA) with apparentlyidentical properties but with separate control mechanisms. Theabsence of major effects of plant growth regulators on the time-courseand rate of nitrate uptake suggests that exogenous regulators,and possibly endogenous phytohormones are of minor importancefor initial nitrate uptake. The differential effect of someregulators on nitrate uptake and root NRA furthermore indicatesthat the processes of uptake and reduction of NO^–₃ arenot obligatory or exclusively coupled to each other.     

Efficiency and regulation of root respiration in a legume: Effects of the N source

A comparison was made of energy metabolism of nodulated N₂ fixing plants and non-nodulated NO₃-fed plants of Lupinus albus L. Growth, N-increment, root respiration (O₂ uptake and CO² production) and the contribution of a SHAM-sensitive oxidative pathway (the alternative pathway) in root respiration were measured. Both growth rate and the rate of N-increment were the same in both series of plants. The rate of root respiration, both O₂ uptake and CO₂ production, and the activity of the SHAM-sensitive pathway were higher in NO₃-fed plants than in N₂ fixing plants. The rate of ATP production in oxidative phosphorylation was computed also to be higher in NO₃-fed plants. It is concluded that both carbohydrate costings and ATP costings for synthesis + maintenance of root material were lower in N₂ fixing than in NO₃-fed plants. The respiratory quotient of root respiration was 1.6 in N₂-fixing plants and 1.4 in NO₃-fed plants. These values were slightly higher than the values calculated on the basis of CO₂ output due to N-assimilation and the experimental values of O₂ uptake, but showed the same trend: highest in N₂ fixing plants. Root respiration of NO₃-fed plants showed a diurnal pattern (both O₂ uptake, CO₂ production and the activity of the SHAM-sensitive pathway), whilst no diurnal variation in root respiration was found in N₂ fixing plants. However, C₂H₂ reduction did show a diurnal rhythm, which is suggested to be related to the diurnal variation in transpiration. Addition of NO₃ to N₂ fixing plants increased the rate of root respiration and the activity of the alternative pathway. This treatment did not decrease C₂H₂ reduction and H₂ evolution within 4 days. Withdrawal of NO₃-supply from NO₃-fed plants decreased the rate of root respiration but had no effect on the relative activity of the alternative pathway. It is suggested that the higher rate of root respiration and the higher activity of the SHAM-sensitive pathway in NO₃-fed plants is due to a larger supply of carbohydrates to the roots, partly due to a better photosynthetic performance of the shoots and partly due to a higher capacity of the roots to attract carbohydrates.     

Direct Measurements of Turgor Pressure Potentials of Guard Cells: II. THE MECHANICAL ADVANTAGE OF SUBSIDIARY CELLS, THE SPANNUNQSPHASE, AND THE OPTIMUM LEAF WATER DEFICIT

Evidence of the mechanical advantage of subsidiary cells wasobtained by simultaneous measurements of turgor pressure potentialsin adjacent subsidiary and guard cells using injection circuitswith two separate needles. In Tradescantia virginiana the mechanicaladvantage approaches two. Using the same technique evidencewas obtained that the Spannungsphase is, in the first place,a turgor relations phenomenon due to the mechanical advantageof epidermal or subsidiary cells. In addition, the evidenceindicated that the elastic properties of guard cell walls mayundergo changes during the Spannungsphase when potassium iontransport commences. During these measurements it was confirmedthat the optimum leaf water deficit for maximum stomatal openingoccurs when the epidermal turgor is near zero. Under these conditionsthe width of the stomatal pore is a function of the turgor pressureof the guard cells, since at zero turgor of the subsidiary cellstheir mechanical advantage has disappeared.