The Carbohydrate Nutrition of Tomato Roots: VII. Sugars, Sugar Phosphates, and Sugar Alcohols as Respiratory Substrates for Excised Roots

The respiratory responses of substrate-depleted excised roottips to a range of sugars, sugar phosphates, and sugar alcoholshave been determined by measuring oxygen uptake by the directmethod of Warburg. Sucrose, dextrose, and laevulose are the only sugars which promotea high level of oxygen uptake. The effects of azide and DNP on the oxygen uptake promoted bysucrose and by dextrose are described. Mannose is a strong inhibitor of respiration. This inhibitionis reversed by the simultaneous addition of those sugars whichalso reverse the growth inhibition caused by mannose. Mannoseinhibits the respiration of sucrose and of glycolytic intermediates.Galactose is slowly respired and does not, even at high concentration,inhibit the respiration of sucrose. The results are discussed in relation to the growth effectsof the sugars tested.     

Respiration-Dependent Utilization of Sugars in Yeasts: a Determinant Role for Sugar Transporters

In many yeast species, including Kluyveromyces lactis, growth on certain sugars (such as galactose, raffinose, and maltose) occurs only under respiratory conditions. If respiration is blocked by inhibitors, mutation, or anaerobiosis, growth does not take place. This apparent dependence on respiration for the utilization of certain sugars has often been suspected to be associated with the mechanism of the sugar uptake step. We hypothesized that in many yeast species, the permease activities for these sugars are not sufficient to ensure the high substrate flow that is necessary for fermentative growth. By introducing additional sugar permease genes, we have obtained K. lactis strains that were capable of growing on galactose and raffinose in the absence of respiration. High dosages of both the permease and maltase genes were indeed necessary for K. lactis cells to grow on maltose in the absence of respiration. These results strongly suggest that the sugar uptake step is the major bottleneck in the fermentative assimilation of certain sugars in K. lactis and probably in many other yeasts.     

Effect of different sugars on flowering ofChenopodium rubrum L. in dependence on the conditions of germination and initial growth

Flowering ofChenopodium rubrum seedlings fed different sugars at a concentration of 0.6 and 0.4 M, reap, during a single inductive cycle was stimulated or inhibited in dependence on the conditions of germination and initial growth. Plants allowed to germinate at alternating temperatures of 28 °C and 5 °C showed a slower initial growth and their development was stimulated by some sugars as compared to controls induced in the absence of sugars. Plants germinated at alternating temperatures of 32 °C and 5 °C exhibited a rapid initial growth and flowering was inhibited after induction in the presence of sugars. On the other hand, development proceeded more rapidly in control plants induced in the absence of sugars after germination at the higher temperature than after germination at the lower one. The differences between the two variants quoted above could be observed also after induction by two 16 h dark cycles. Glucose and sucrose were most effective in stimulating flowering under appropriate conditions of germination. Fructose was less effective and the action of maltose was very weak. Xylose, ribose and galactose were innocuous, while arabinose, glucoso-6-phosphate and mannitol were toxic to the plants. The sugars inhibited root growth in all cases and led to an increase in starch accumulation in the underground and overground plant organs. At a concentration of 0.6 M they mostly inhibited the length of the cotyledons and, especially, of the first leaf; at a concentration of 0.4 M growth of the overground organs was stimulated. The results are discussed with respect to the possible ohanges in photoperiodic sensitivity brought about by the rate of initial growth.     

The role of microbial signals in plant growth and development

Plant growth and development involves a tight coordination of the spatial and temporal organization of cell division, cell expansion and cell differentiation. Orchestration of these events requires the exchange of signaling molecules between the root and shoot, which can be affected by both biotic and abiotic factors. The interactions that occur between plants and their associated microorganisms have long been of interest, as knowledge of these processes could lead to the development of novel agricultural applications. Plants produce a wide range of organic compounds including sugars, organic acids and vitamins, which can be used as nutrients or signals by microbial populations. On the other hand, microorganisms release phytohormones, small molecules or volatile compounds, which may act directly or indirectly to activate plant immunity or regulate plant growth and morphogenesis. In this review, we focus on recent developments in the identification of signals from free-living bacteria and fungi that interact with plants in a beneficial way. Evidence has accumulated indicating that classic plant signals such as auxins and cytokinins can be produced by microorganisms to efficiently colonize the root and modulate root system architecture. Other classes of signals, including N-acyl-L-homoserine lactones, which are used by bacteria for cell-to-cell communication, can be perceived by plants to modulate gene expression, metabolism and growth. Finally, we discuss the role played by volatile organic compounds released by certain plant growth-promoting rhizobacteria in plant immunity and developmental processes. The picture that emerges is one in which plants and microbes communicate themselves through transkingdom signaling systems involving classic and novel signals.Key words: Arabidopsis, alkamides, auxins, quorum-sensing, cytokinins     

The Relationship between Cyanide-Resistant Root Respiration and the Storage of Sugars in the Taproot in Daucus carota L.

Dry weight increase, root respiration, photosynthesis, and shootdark respiration were measured prior to and during the developmentof the taproot of carrot (Daucus carota L., var. AmsterdamseBak) which stores sucrose and reducing sugars. Before storageof sugars in the taproot started, root respiration was highand up to 50% inhibited by salicyl hydroxamic acid (SHAM), indicatinga high activity of the alternative pathway. The onset of sugar storage in the taproot coincided with a sharpdecrease in alternative pathway activity. When sugars were storedin the taproot, the alternative pathway in the taproot was notoperative, although present. It was concluded that the alternative pathway only consumessugars that are not used for energy production in growth ormaintenance processes, for carbon skeletons, osmoregulationor storage, in accordance with the energy overflow model. Apositive quantitative relationship exists between the storageof sugars in the taproot and the efficiency of root respiration.After day 32 the amount of daily produced carbohydrates transportedto the root system increased and these sugars were stored inthe taproot. In this period less sugars were invested in structuralshoot growth. In the present investigation no positive influence from thedeveloping sink is found on photosynthetic rate or net assimilationrate. Both rates decrease with increasing age of the plants,although increasing amounts of sugars were stored in the taproot.     

Sensitivity of apple seed germination to light and some growth regulators

Experiments were aimed at checking whether the non-stratified apple embryos are sensitive to growth factors and light. The effect of growth regulators on light sensitivity was also studied. The stimulating or inhibiting effect of GA3, benzyladenine and IAA or coumarin on the dynamics of apple embryo germination was demonstrated. Seasonal fluctuations of the effect of the growth factors were noted. The stimulating effect of light on the germination of non-stratified apple embryos was demonstrated. This effect is independent of season and seems to be independent of the effects of growth regulators.     

The Kluyver effect revisited

Yeast species can grow on various sugars. However, in many cases the growth on certain sugars (especially oligosaccharides) occurs only under aerobic conditions, and not in anaerobiosis or in the absence of respiration. Fermentation is blocked under these conditions. This apparent dependence of sugar utilization on the respiration has been called Kluyver effect, and such 'respiration-dependent' species are called Kluyver effect positive. A yeast may be Kluyver effect positive for some sugars and not for others. The physiological meaning and the molecular basis of the phenomenon are not clear. It has recently been reported that Kluyveromyces lactis, which is Kluyver effect positive for galactose and a few other sugars, could be converted into a Kluyver effect-negative form by introduction of relevant sugar transporter genes. Such results offer for the first time a direct support to the hypothesis that the immediate cause of the Kluyver effect may be the low level of sugar transporter activities which is not sufficient to sustain the high substrate flow necessary for fermentative growth, whereas the energy-efficient respiratory growth does not require a high rate of sugar uptake. We examined to what extent this sugar transporter theory of the Kluyver effect can be generalized.     

Effect of gibberellic acid on epicotyl growth and carbohydrate distribution in derooted Pisum sativum cuttings with or without cotyledons

The effect of gibberellic acid (GA) on subhook growth in derooted cuttings of pea ( Pisum sativum L. cv. Alaska) grown in the dark was studied in relation to the distribution of sugar-related compounds in the epicotyl and cotyledons. GA stimulated subhook growth of cuttings with or without cotyledons. In cuttings with cotyledons, the net inflow of sugar-related compounds (soluble sugars, starch, cell wall polysaccharides and sugars consumed by respiration) to the epicoiyl balanced with the net outflow from the cotyledons. GA stimulated the net inflow of sugar-related compounds to the epicotyl and the net outflow from cotyledons. Among these compounds, GA substantially increased the amount of soluble sugars, starch and cell wall polysaccharides in the subhook. In cuttings without cotyledons, on the other hand, the net inflow of sugar-related compounds to the subhook almost balanced with the net outflow from the epicotyl below the subhook. GA stimulated the net inflow of sugar-related compounds to the subhook and the net outflow from the epicotyl below the subhook. Among these compounds, GA substantially increased the amount of soluble sugars and cell wall polysaccharides in the subhook. These results suggest that GA stimulates an increase in the net inflow of sugar-related compounds to the subhook, thereby preventing an increase in osmotic potential and stimulating cell wall polysaccharide synthesis, when pea subhook growth is stimulated.     

On the lateral growth correlations exemplified by petioles and axillaries ofPrisum cotyledons

The cotyledonary petioles inPisum sativum developing at the start of germination, as well as the cotylaries growing out after decapitation of thePisuin seedlings manifest very often the so-called lateral correlations, since one of them suppresses the opposite primordium when the growth inhibition in the plant coinciding apparently with the growth promotion results necessarily in a normal development only of one of these originally equal primordia. These correlations also seem to be mediated by auxin, being induced by gravity or by corresponding cotyledon and root products, which can be supported by the use of exogenous auxins.     

Regulation of the biomass and activity of soil microorganisms by microfauna

Microcosm experiments showed that the microbial biomass and the respiration activity in soil were regulated by nematodes. Depending on nematode number and plant residue composition, the trophic activity of nematodes can either stimulate or inhibit microbial growth and respiration as compared to soil containing no nematodes. The stimulating effect was observed when nitrogen-free (starch) or low-nitrogen (wheat straw, C : N = 87) organic substrates were applied. Inhibition occurred when a substrate rich in nitrogen (alfalfa meal, C : N = 28) was decomposed and the nematode population exceeded the naturally occurring level. A conceptual model was developed to describe trophic regulation by microfauna (nematodes) of the microbial productivity and respiration ctivity and decomposition of not readily decomposable organic matter in soil. The stimulating and inhibiting influence of microfauna on soil microorganisms was not a linear function of the rate of microbial consumption by nematodes. These effects are largely associated with the induced change in the physiological state of microorganisms rather than with the mobilization of biogenic elements from the decomposed microbial biomass.     

Regulation of the biomass and activity of soil microorganisms by microfauna

Microcosm experiments showed that the microbial biomass and the respiration activity in soil were regulated by nematodes. Depending on nematode number and plant residue composition, the trophic activity of nematodes can either stimulate or inhibit microbial growth and respiration as compared to soil containing no nematodes. The stimulating effect was observed when nitrogen-free (starch) or low-nitrogen (wheat straw, C:N = 87) organic substrates were applied. Inhibition occurred when a substrate rich in nitrogen (alfalfa meal, C:N = 28) was decomposed and the nematode population exceeded the naturally occurring level. A conceptual model was developed to describe trophic regulation by microfauna (nematodes) of the microbial productivity and respiration activity and decomposition of not readily decomposable organic matter in soil. The stimulating and inhibiting influence of microfauna on soil microorganisms was not a linear function of the rate of microbial consumption by nematodes. These effects are largely associated with the induced change in the physiological state of microorganisms rather than with the mobilization of biogenic elements from the decomposed microbial biomass.     

The physiological basis of seed dormancy in Avena fatua VI. Respiration and the stimulation of germination by ethanol

Ethanol induced germination in several partly after-ripened dormant lines of Avena fatua L. The dose-response curves for the stimulation of germination and for oxygen uptake were similar, indicating that ethanol may stimulate germination by promoting oxygen uptake. A time-sequence study showed that ethanol stimulated oxygen uptake by as much as 70% prior to the first visible signs of germination. A similar methanol treatment failed to induce germination or significantly elevate oxygen uptake, indicating that the promotive effects of ethanol are not common to all alcohols. The stimulation of both germination and oxygen uptake by ethanol was not inhibited significantly by salicylhydroxamic acid, an inhibitor of alternative respiration. Thus, stimulation of germination and oxygen uptake by ethanol does not require the operation of the alternative pathway of respiration. Similarly, the stimulation of germination and oxygen uptake by ethanol were not inhibited by sodium azide, an inhibitor of cytochrome-mediated respiration. However, both germination and oxygen uptake were prevented when salicylhydroxamic acid and sodium azide were administered together. Thus, stimulation of these events by ethanol requires only the operation of one or other of these pathways of respiration; a specific requirement for the operation of the alternative pathway of respiration does not exist. The function of ethanol as a promoter of respiration is discussed with reference to dormancy and involvement of the Krebs cycle.     

Enhancement of ethylene biosynthesis and germination with thidiazuron and some selected auxins in Striga asiatica seeds

Witchweed [ Striga asiatica (L.) Kuntze], an economically important parasitic weed on several poaceous crops, is difficult to control. In nature, germination and subsequent morphogenesis of Striga are cued to specific host-derived chemical signals. Seeds (approximately 2.4 mg) treated with thidiazuron (TDZ) or the auxins 2,4-dichlorophenoxy-acetic acid (2,4-D), 1-naphthalene acetic acid (NAA), or 2-(4-chloro- o -tolyloxy) propionic acid (MCPP) produced little ethylene (66-138 nl l⁻¹). Combinations of TDZ with the auxins increased ethylene production by 4- to 18-fold. Ethylene production was strongly inhibited (86–92%) by aminoethoxyvinylglycine (AVG), inhibitor of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase. Ethylene evolved from seeds treated with TDZ in combination with 2,4-D increased after a lag period and was promoted by a pretreatment in 2,4-D. TDZ or any of the auxins, at the rates tested, effected negligible to low levels of germination (0 to 16%), whereas mixtures of TDZ with the above auxins stimulated 38 to 84% germination. Test solutions containing TDZ and indole-3-acetic acid (IAA) were, however, less effective. TDZ/auxin-induced germination was inhibited by AVG and the ethylene action inhibitor silver thiosulfate (STS). The inhibitory effect of the former was reversed by treatment with ACC. In vitro studies revealed negligible germination (< 1%) on control medium. Seeds germinating on media containing TDZ alone developed into seedlings with distinct shoots and rudimentary roots. Seeds germinating on media containing 2,4-D, irrespective of TDZ concentration, were induced to form calli. The results are consistent with a model in which both germination and subsequent morphogenesis in Striga are associated with exogenous and endogenous phytohormones.     

Tomato pollen respiration in relation to in vitro germination and pollen tube growth under favourable and stress-inducing temperatures

Tomato pollen germination, pollen tube growth and respiratory activity were recorded during incubation in a liquid medium for 7 h over a temperature range of 15–35°C. Although the initial rate of respiration was highest at 30°C, both at 30°C and 35°C respiration decreased after the first hour of incubation due to high temperature impairment of germination and pollen tube growth. The total per cent germination of pollen over the 7-h period was maximal at 15°C whereas pollen tube length was maximal at 25°C. Although the production of CO₂ measured at hourly intervals throughout the incubation period did not correlate to a statistically significant level with either the per cent pollen germination or the length of the pollen tubes alone, nevertheless from 2 h after the start of incubation, it closely correlated with the values for germination × pollen tube length, indicating that the respiratory activity of tomato pollen at a given time is a function of both the per cent germination and the pollen tube growth. We suggest therefore that the rate of respiration might be preferable to a simple germination test for the assessment of pollen germination ability since it expresses not only the pollen germination potential but also the growth vigour of the pollen tubes. In addition, where in vitro tests are designed to assess pollen germination–temperature interactions, they should employ a long incubation period (e.g. 7 h) to permit differences in sensitivity to temperature to be observed.     

Biochemical studies of the regeneration of the root cuttings ofClerodendrum viscosum Vent

The contents of reducing sugars, proteins and ascorbic acid and the activities of protease, peroxidase, catalase, indole acetic acid oxidase, ascorbic acid oxidase, succinic dehydrogenase and the rate of respiration were investigated in regenerating and non-regenerating parts ofClerodendrum viscosum Vent. root cuttings. All the above variables have registered higher in the regenerating parts, particularly at the distal end where roots appear, than in the non-regenerating parts of root cuttings. The contents of metabolites and the activities of enzymes in the presence of endogenous cytokinins at the proximal end and auxins at the distal end, were compared and discussed in relation to the initiation of shoots and roots at two ends of root cuttings. In general it was concluded that interacting hormonal and nutritional factors must be appropriate to initiate shoots or roots in the root cuttings.     

Possible Involvement of the Alternative Respiration System in the Ethylene-stimulated Germination of Cocklebur Seeds

Respiration of nondormant upper cocklebur (Xanthium pensylvanicum Wallr.) seeds was enhanced by exogenous C₂H₄, proportionally to the concentration of C₂H₄ and the duration of presoaking of the seeds. Benzohydroxamic acid (BHM) and salicylhydroxamic acid (SHM), inhibitors of alternative respiration, inhibited both the germination of nondormant lower cocklebur seeds and the respiration of the upper seeds presoaked for periods of 12 to 30 hours. Both the growth and respiration of axial and cotyledonary tissues were also inhibited by BHM. Moreover, BHM inhibited both the C₂H₄-induced germination of the upper seeds and their C₂H₄-stimulated respiration; the inhibition occurred only with concomitant addition of C₂H₄ and BHM. The respiration of seeds with a secondary dormancy induced by presoaking for prolonged periods was markedly stimulated by C₂H₄ but not suppressed by BHM. It was suggested that the alternative respiration system may be involved in the normal germination process of cocklebur seeds, secondary dormancy may result from its inactivation, and C₂H₄ may exert its germination-promoting action by stimulating the alternative respiration. The effects of BHM and SHM can suggest but not prove the involvement of the alternative respiration in seed germination.     

Effects of salicylhydroxamate on respiration, seed germination and seedling growth in Avena fatua

The effects of inhibitors of alternative respiration [salicylhydroxamate (SHAM) and propyl gallate (PG)] on germination, seedling growth and O₂ uptake in Avena fatua L. (wild oats) were studied. SHAM did not inhibit germination or O₂ uptake prior to germination. SHAM-sensitive (alternative) respiration, therefore, cannot be a pre-requisite for germination. Following germination, both chemicals inhibited seedling growth with the root being more susceptible than the shoot. SHAM concentrations that inhibited root growth by 90 to 95%, inhibited O₂ uptake of 1 cm root apices by less than 15%. While sodium azide (a cytochrome-oxidase inhibitor; 1 m M ) alone inhibited O₂ uptake by only 40 to 50%, in the simultaneous presence of SHAM (or PG), O₂ uptake was inhibited by 90 to 99%. Thus: 1) respiration of wild oat seedling root apices is predominantly cytochrome-mediated and incomplete inhibition of O₂ uptake in the presence of azide alone is due to diversion of electrons to the alternative pathway and 2) even though these roots have little alternative respiration, they maintain the capacity to support a much greater flux of electrons via this path way. SHAM and PG at concentrations (0.05 to 0.4 m M ) which inhibited O₂ uptake significantly in the presence (but not in the absence) of azide had little effect on root growth suggesting that an effect(s) other than that on respiration is involved in the inhibition of root growth at higher concentrations. The effect of SHAM on wild oat root growth is not selective as it also inhibits growth of a number of crop species.     

The physiological basis of seed dormancy in Avena fatua IV. Alternative respiration and nitrogenous compounds

Sodium nitrate and nitrite induced germination in a partly after-ripened dormant line of Avena fatua L. The dose-response curves for their stimulation of germination and oxygen uptake were similar, indicating that these compounds may stimulate germination by promoting oxygen uptake. Time-sequence studies showed that nitrate and nitrite stimulated oxygen uptake as much as 70% prior to the first signs of germination. A similar ammonium chloride treatment failed to induce germination or elevate the rate of oxygen uptake, indicating that nitrate and nitrite may promote these events by acting as electron acceptors. The stimulation of germination and oxygen uptake by nitrate and nitrite were not significantly inhibited by salicylhydroxamic acid, an inhibitor of alternative respiration. Thus, stimulation of germination and oxygen uptake by nitrate and nitrite do not require the operation of the alternative pathway of respiration. The stimulation of germination and oxygen uptake by nitrate and nitrite were not inhibited by sodium azide, an inhibitor of cytochrome-mediated respiration; however both germination and oxygen uptake were prevented when salicylhydroxamic acid and sodium azide were applied together. Thus, stimulation of germination and oxygen uptake by nitrate and nitrite require the operation of only one or other of the pathways of respiration; however a specific requirement for the operation of the alternative pathway of respiration does not exist. The function of these compounds as promotors of respiration is discussed with reference to dormancy and the involvement of the Krebs cycle.