Effects of Humic Acid on Protein Synthesis and Ion Uptake in Beet Discs

Humic acids stimulate the development of phosphate uptake-capacityin beet discs during ageing under aseptic conditions withoutaffecting phosphate uptake per se. Chloride uptake is inhibitedby humic acids as is the development of chloride uptake capacity.The uptake of proline and leuoine is not influenced by ageingin humic acid. While measurements of the incorporation of theseamino acids into the sub-cellular particles of discs indicatethat humic acid does not affect protein synthesis in general,the stimulation of invertase development during ageing doesshow a definite effect on some aspect of protein synthesis.     

Comparison of the effects of auxin and fusicoccin on phosphate absorption by aged potato tuber discs

Auxin (IAA, 5 × 10⁻⁵ M ) partially prevents the increase in the rate of phosphate uptake during ageing of potato tuber discs ( Solatium tuberosum L. cv. Bintje), whereas fusicoccin (FC, 10⁻⁵ M) stimulates it. After the development of enhanced phosphate transport capacity, the response to fusicoccin is greater than with fresh discs. Complementary experiments on K⁺ (⁸⁶Rb) absorption show that FC also slightly enhances the rate of K⁺ uptake, while IAA has no much effect. It is suggested that IAA acts specifically on the development of a mechanism which occurs during the ageing period, while FC action may be more directly linked to the system of phosphate transport itself.     

Occurrence of Localized Intense Cation Uptake Sites in the Vascular Rings of Red Beet Storage Tissue 2

Development and decline of cation uptake capacity in discs takenfrom the vascular and parenchyma rings of storage tissue ofred table beet (Beta vulgaris L.) were observed during 12 dof ageing. Uptake capacity for Na⁺ and Rb⁺ showed a steady risereaching maximums by the fourth to fifth days of ageing. Thereafter,there was a steady decline in the uptake rates. Vascular ringtissues were able to develop a greater uptake capacity for bothNa⁺ and Rb⁺ than the tissues of parenchyma rings. This difference,which was more pronounced for Rb⁺ than for Na⁺ uptake, is attributedto a combination of variations in cell density and differencesin the acquisition and retention of the cation uptake capacity.Respiration of tissue discs showed no significant rise duringageing, nor were there significant differences in the respirationof vascular and parenchyma tissues. Vascular tissues containedsignificantly more betacyanin than parenchyma tissues; and theyretained their pigment, as well as their acquired cation uptakecapacity, for a longer period during the ageing process. Key words: Cation uptake, Red beet, Vascular rings, Ageing     

The influence of growth regulating substances on the development of enhanced metabolic rates in thin slices of beetroot storage tissue

Freshly cut disks of beetroot tissue develop high rates of respiration, uptake of phosphate and activity of the enzyme invertase after having been washed for 18 hours in 0.01 m potassium maleate.Incubation of the disks in solutions of indole-3-acetic acid or kinetin completely prevented the development of the higher activities in all 3 systems assayed, while incubation in gibberellic acid had no inhibitory effect. Using a series of synthetic plant growth regulating compounds it was possible to establish that there was no correlation between the activity of the compound as an auxin and the ability of the compound to prevent the development of the enhanced rates of metabolism.     

Phosphate uptake and utilization by bacteria and algae

Bacterial uptake of inorganic phosphate (closely investigated in Escherichia coli) is maintained by two different uptake systems. One (Pst system) is P_i-repressible and used in situations of phosphorus deficiency. The other system (Pit system) is constitutive. The Pit system also takes part in the phosphate exchange process where orthophosphate is continuously exchanged between the cell and the surrounding medium.Algal uptake mechanisms are less known. The uptake capacity increases during starvation but no clearly defined transport systems have been described. Uptake capacity seems to be regulated by internal phosphorus pools, e.g., polyphosphates. In mixed algal and bacterial populations, bacteria generally seem to be more efficient in utilizing low phosphate concentrations. The second half of this paper discusses how bacteria and algae can share limiting amounts of phosphate provided that the bacteria have pronouncedly higher affinity for phosphate. Part of the solution to this problem may be that bacteria are energy-limited rather than phosphate-limited and dependent on algal organic exudates for their energy supply.The possible phosphate exchange mechanism so convincingly demonstrated in Escherichia coli is here suggested to play a key role for the flux of phosphorus between bacteria and algae. Such a mechanism can also be used to explain the rapid phosphate exchange between the particulate and the dissolved phase which always occurs in short-term ³²P-uptake experiments in lake waters.     

Further evidence of oxygen diffusion as the determining factor in the relation between disk thickness and respiration of potato tissue

The effect of oxygen tension above atmospheric pO₂ on the development of respiratory capacity in potato disks has been examined. Raising the oxygen tension of the aqueous environment to 40% during the aging of 2.0 mm or 3.0 mm thick disks at 25° progressively increased the respiration rate of the tissue as shown by subsequent assay in 100% oxygen. Disks 3.0 mm thick showed a greater response to increased pO₂ than did 2.0 mm disks. A comparison of center 1.0 mm sections excised from 3.0 mm disks after aging, showed that the respiration rate of internal tissue from disks aged in high pO₂ was approximately 40% greater than such tissue aged with atmospheric pO₂. The characteristic inverse relationship between respiration rate and thickness in aged disks can be modified from a concave-downwards curve to a convex-downwards curve by pretreating the tissue with increased pO₂, thus indicating that raising the pO₂ during aging can increase the thickness threshold at which the transition from tissue manifesting the respiratory characteristics of thin disks to that manifesting the characteristics of thick disks, occurs. Similarly increased pO₂ during aging can modify the hyperbolic relationship obtaining between pretreatment temperature in the range 10° to 25° and respiratory capacity of aged 3.0 mm disks, to approximate to the linear relationship observed with 0.75 mm disks. It is concluded that the development of respiratory capacity in disks between 0.75 mm and 3.0 mm thick is restricted by oxygen dificiency and that the characteristic inverse relationship between respiration rate and thickness in aged disks is largely attributable to this factor, the influence of which is discernible both on the development of respiratory capacity and on its subsequent assay.     

Response to trisodium phosphate treatment of Salmonella Chester attached to fresh-cut green pepper slices

A laboratory model using green pepper disks was developed to investigate the attachment of Salmonella Chester on plant tissue and to evaluate the effectiveness of sanitizer agents in inactivating attached bacteria on fruits. Pepper disks (14 mm in diam, and 3-4 mm in thickness) were immersed in a bacterial suspension containing 1.5 x 107 cfu x mL(-1) of S. Chester for 30 s and subsequently air-dried at room temperature for 10 min. Approximately 30% of the bacteria retained on the disk after immersion were firmly attached and could not be removed by two washes and agitation. A positive correlation was observed between the number of bacteria attached and the concentration of bacteria in the suspension. Population studies and scanning electron microscopic examinations revealed that attachment of S. Chester on pepper disks occurred mainly on the surfaces of injured (cut) tissue but rarely on the unbroken skin. When inoculated disks were treated with 3% to 12% (w/v) of trisodium phosphate (TSP) at pH 12.3 for 5 min, the population of bacteria on the disk was reduced by 10- to 100-fold. A small portion (0.7% to 7.1%) of bacteria attached to the disk were either resistant to or protected from the TSP treatment. When the pH of TSP solution was reduced from 12.3 to 4.5, the effectiveness of TSP in inactivating S. Chester on pepper disks was reduced by 26%. This study shows that surfaces of injured fruit tissue are the principal sites for bacterial attachment, and a small portion of the bacteria attached to the tissue are resistant to the sanitizer treatment. Avoiding mechanical injuries to fresh fruits during and after harvest would reduce the chance of pathogen attachment and contamination on green pepper and fruits of similar nature.     

The Effect of Vanadate on Phosphate Transport in Aged Potato Tuber Tissue 2Solarium tuberosum)

The effect of orthovanadate on the uptake of phosphate by agedpotato tuber tissue was investigated to study the relationshipwith plasma membrane ATPase activity. Vanadate inhibited therate of phosphate uptake by aged discs with a maximum effectat 500 µM (58% inhibition). When vanadate was added tothe ageing medium for 24 h, the subsequent rate of phosphateuptake was also markedly decreased (68% inhibition). The resultsshow that the inhibition by vanadate was not due to enhancedleakage of phosphate nor to a non-specific toxic effect. Furthermore,complementary experiments with erythrosin B and molybdate wereconsistent with the hypothesis that vanadate acts specificallyon the plasma membrane ATPase and that this enzyme is involvedin maintaining the driving force for active uptake of phosphate(via co-transport with protons) by storage cells of potato tubers. Key words: Proton-phosphate co-transport, vanadate, plasma membrane ATPase, unloading     

Changes in Sensitivity to Inhibitors of Disks of Storage Tissue

The respiratory sensitivity to cyanide of disks of sugar-beet,red beet, carrot, swede, and potato maintained at differenttemperatures over a prolonged period has been determined manometrically,and the effects of sodium azide and carbon monoxide examinedon sugar-beet disks. The normally high sensitivity to cyanideof freshly cut disks falls more or less rapidly depending onthe temperature of pretreatment until the oxygen uptake is practicallyunaffected by the inhibitor. Subsequently the sensitivity redevelopsbut a brief period during which cyanide exercises a stimulatoryeffect may be interposed. With beet and carrot disks containingcambial cells there is in the early stages of the lifespan aperiod of increasing sensitivity to cyanide. It is concludedthat there is a characteristic difference in cyanide-sensitivitybetween growing and non-growing tissue. A discussion of underlyingtheory is added.     

Invertase Development in Storage Tissue 4Beta vulgaris; its Nature, Extent, and Location

The development of invertase activity in storage tissue disksof Beta vulgaris during ageing under aseptic conditions hasbeen studied. The invertase is formed initially in the cellwall and subsequently appears in the cytoplasm. In disks upto 1.0 mm thick, invertase is formed throughout the tissue,but in thicker disks only limited activity appears in the interior.In disks up to 1.0 mm thick, treatment with ethyl acetate priorto enzyme assay increases the measured activity by rendering‘inaccessible’ soluble invertase available to sucrosein the assay medium. In thicker disks, pretreatment with ethylacetate also increases the measured invertase activity by facilitatingsucrose penetration to the centre of the tissue. Osmotic shockaffects invertase activity in the same manner as ethyl acetatetreatment. The significance of these results is discussed inrelation to the induction and development of invertase activityin the disk and in the cell.     

Influence of auxin on phosphate absorption and metabolism of phosphorylated compounds in aged potato tuber discs (Solanum tuberosum)

Ageing of potato tuber discs markedly increases the rate of phosphate uptake. This increase is partially prevented by the presence of indoleacetic acid (IAA: 50 μ M ) in the ageing medium. ³²P distribution among the various phosphorylated fractions (P₁, organic soluble phosphate, acid-insoluble phosphate) was carried out after 24 h of ageing in the presence of IAA. An equal inhibition of the labelling rates of each of the different fractions is induced by the hormone. No important effect on respiration and ATP content was observed. Moreover, IAA neither changes the total phospholipid content nor the relative distribution of ³²P between the components. These results support the hypothesis that IAA acts specifically on the development of uptake mechanisms during the ageing period.     

The uptake of inorganic nutrients by heterotrophic bacteria

It is now well known that heterotrophic bacteria account for a large portion of total uptake of both phosphate (60% median) and ammonium (30% median) in freshwaters and marine environments. Less clear are the factors controlling relative uptake by bacteria, and the consequences of this uptake on the plankton community and biogeochemical processes, e.g., new production. Some of the variation in reported inorganic nutrient uptake by bacteria is undoubtedly due to methodological problems, but even so, uptake would be expected to vary because of variation in several parameters, perhaps the most interesting being dissolved organic matter. Uptake of ammonium by bacteria is very low whereas uptake of dissolved free amino acids (DFAA) is high in eutrophic estuaries (the Delaware Bay and Chesapeake Bay). The concentrations and turnover of DFAA are insufficient, however, in oligotrophic oceans where bacteria turn to ammonium and nitrate, although the latter only as a last resort. I argue here that high uptake of dissolved organic carbon, which has been questioned, is necessary to balance the measured uptake of dissolved inorganic nitrogen (DIN) in seawater culture experiments. What is problematic is that this DIN uptake exceeds bacterial biomass production. One possibility is that bacteria excrete dissolved organic nitrogen (DON). A recent study offers some support for this hypothesis. Lysis by viruses would also release DON.While ammonium uptake by heterotrophic bacteria has been hypothesized to affect phytoplankton community structure, other impacts on the phytoplankton and biomass production (both total and new) are less clear and need further work. Also, even though bacteria account for a very large fraction of phosphate uptake, how this helps to structure the plankton community has not been examined. What is clear is that the interactions between bacterial and phytoplankton uptake of inorganic nutrients are more complicated than simple competition.     

Influence of Abscisic Acid on Phosphorus Metabolism and Respiration in Potato Tuber Discs. Comparison with the Cycloheximide Effect

Ageing of discs of potato tuber by incubation in an aerated medium, produces an increase in the rates of respiration and of phosphate uptake. The presence of cycloheximide (CHM) or abscisic acid (ABA) in the uptake medium, does not change uptake by fresh tissue over 3–4 h. On the other hand, CHM causes an inhibition of the rate of uptake by aged tissue although ABA does not. The addition of CHM or ABA to the ageing medium, prevents totally (CHM) or only partially (ABA) the increase in phosphate uptake. The analysis of ³²P-incorporation into the various phosphorylated fractions after 24 h of ageing with CHM or ABA show that CHM induces a large inhibition of the rate of uptake with an almost complete inhibition of ³²P-incorporation into the various phosphorylated fractions. By contrast, ABA produces equal inhibition of ³²P labelling of all fractions including all acid-soluble components. CHM prevents the increase in the rate of respiration, whereas ABA causes a slight stimulation. In both cases, no important effect on ATP content was observed. These results are discussed in terms of a comparison of ABA and CHM actions. They lead to the hypothesis of a specific effect of ABA on the development of the uptake mechanism.     

Telomeres as biomarkers for ageing and age-related diseases

Telomeres in telomerase-negative cells shorten during DNA replication in vitro due to numerous causes including the inability of DNA polymerases to fully copy the lagging strand, DNA end processing and random damage, often caused by oxidative stress. Short telomeres activate replicative senescence, an irreversible cell cycle arrest. Thus, telomere length is an indicator of replicative history, of the probability of cell senescence, and of the cumulative history of oxidative stress. Telomeres in most human cells shorten during ageing in vivo as well, suggesting that telomere length could be a biomarker of ageing and age-related morbidity. There are two distinct possibilities: First, in a tissue-specific fashion, short telomeres might indicate senescence of (stem) cells, and this might contribute to age-related functional attenuation in this tissue. Second, short telomeres in one tissue might cause systemic effects or might simply indicate a history of high stress and damage in the individual and could thus act as risk markers for age-related disease residing in a completely different tissue. In recent years, data have been published to support both approaches, and we will review these. While they together paint a fairly promising picture, it needs to be pointed out that until now most of the evidence is correlative, that much of it comes from underpowered studies, and that causal evidence for essential pathways, for instance for the impact of cell senescence on tissue ageing in vivo, is still very weak.     

The Influence of Internal Ion Concentration on Potassium Accumulation and Salt Respiration of Red Beet Root Tissue

It is shown that during a period of washing in serated distilledwater disks of Red Beet root tissue acquire the capacity toabsorb potassium rapidly. Subsequently the rate of accumulationof this ion is closely related to the internal salt-contentof the material. On the other hand, whilst the level of saltrespiration increases during the preliminary washing, it isnot influenced significantly by the internal potassium concentration.The implications of these observations are discussed in relationto a possible mechanism of mineral salt absorption.     

Activity of enzymes involved in lignin biosynthesis in swede root disks

The activity of nine enzymes involved in the biosynthesis of lignin precursors has been studied during the ageing of swede root disks in the presence and absence of ethylene. Peroxidase, aromatic alcohol dehydrogenase and phenylalanine transaminase show very little change in activity during ageing under either ageing condition. O-methyl transferase, shikimate dehydrogenase and ferulyl CoA reductase show only a 2–3 fold increase on ageing and are relatively insensitive to ethylene treatment. A third group (comprising phenylalanine ammonia lyase, cinnamic acid-4-hydroxylase and hydroxycinnamate CoA ligase) show 20–30 fold increase on ageing and are most sensitive to ethylene treatment. Phenylalanine ammonia lyase and cinnamic acid-4-hydroxylase behave very similarly in respect of their time course of ageing and in their responses to metabolic inhibitors such as cycloheximide, puromycin and actinomycin D. In addition the properties of the O-methyl transferase of swede root tissue are described.     

Inhibition of the Development of a Cation Accuniulatory System and of Tris-induced Uptake in Storage Tissues by N6-benzyladenine and Kinetin

The effect of N⁶-benzyladenine (BA) and kinetin on cation uptake in disks of beetroot (Beta vulgaris L.) and swede (Brassica napobrassica Mill.) tissue was measured in aerated solutions, containing 1 mM NaCl or 1 mM KCl, and in the presence or absence of trishydroxymethyl ammo methane (tris) buffer at pH 8. This investigation followed a suggestion that the immediate activation of cation uptake in freshly sliced beetroot disks by tris (the tris-effect) may depend on stimulation of hexokinase activity. BA, a competitive inhibitor of hexokinase, caused a complete inhibition of tris stimulated cation uptake in beetroot disks, but the effect was delayed and preceded by a 3–4 hour period of promoted cation uptake. Generally, the effects of BA and kinetin were identical. Use of ¹⁴C-kinetin indicated a rapid incorporation of kinetin into the tissue. BA and kinetin also prevented the development of a cation uptake capability during the aging of the beetroot disks. Swede disks which do not show a tris-effcct and no lag phase in development of cation uptake capability were not affected by BA and kinetin treatments. Mechanisms of tris induced cation uptake are discussed with emphasis on its role as a proton acceptor.     

Response of Thiobacillus ferrooxidans to phosphate limitation

Abstract: Phosphate ion is an essential nutrient for all cells. Consequently, starvation for this component may constitute a stressing condition which affects the bioleaching capacity of the biomining microorganisms. Therefore, we have studied the manner in which the chemolithotroph Thiobacillus ferrooxidans responds to phosphate limitation. Under these circumstances the bacteria reduced its growth rate, capacity to oxidize ferrous iron and to fix CO,. Concomitant with these changes, the cells showed an increased synthesis of several proteins, some of which were exclusively synthesized during phosphate starvation. When intact cells grown in the absence of phosphate were labelled with ¹²⁵I, several proteins were iodinated in addition of those observed in control cells, suggesting that the lack of phosphate induces some proteins located in the membranes or the periplasmic space of the bacteria. It is expected that by measuring the levels of expression of some of the proteins induced by the shortage of phosphorus, it might be possible to estimate in situ the relative physiological condition of the bacteria in a given bioleaching operation.     

Some properties of pyruvate and 2-oxoglutarate oxidation by blowfly flight-muscle mitochondria

1. High rates of state 3 pyruvate oxidation are dependent on high concentrations of inorganic phosphate and a predominance of ADP in the intramitochondrial pool of adenine nucleotides. The latter requirement is most marked at alkaline pH values, where ATP is profoundly inhibitory. 2. Addition of CaCl(2) during state 4, state 3 (Chance & Williams, 1955) or uncoupled pyruvate oxidation causes a marked inhibition in the rate of oxygen uptake when low concentrations of mitochondria are employed, but may lead to an enhancement of state 4 oxygen uptake when very high concentrations of mitochondria are used. 3. These properties are consistent with the kinetics of the NAD-linked isocitrate dehydrogenase (EC 1.1.1.41) from this tissue, which is activated by isocitrate, citrate, ADP, phosphate and H(+) ions, and inhibited by ATP, NADH and Ca(2+). 4. Studies of the redox state of NAD and cytochrome c show that addition of ADP during pyruvate oxidation causes a slight reduction, whereas addition during glycerol phosphate oxidation causes a ;classical' oxidation. Nevertheless, it is concluded that pyruvate oxidation is probably limited by the respiratory chain in state 4 and by the NAD-linked isocitrate dehydrogenase in state 3. 5. The oxidation of 2-oxoglutarate by swollen mitochondria is also stimulated by high concentrations of ADP and phosphate, and is not uncoupled by arsenate.