Ammonium and nitrate uptake rates and rhizosphere pH in non-mycorrhizal roots of Norway spruce [Picea abies (L.) Karst.]

Summary Relationships between root zone temperature, concentrations and uptake rates of NH ₄ ⁺ and NO ₃ ^– were studied in non-mycorrhizal roots of 4-year-old Norway spruce under controlled environmental conditions. Additionally, in a forest stand NH ₄ ⁺ and NO ₃ ^– uptake rates along the root axis and changes in the rhizosphere pH were measured. In the concentration (C_min) range of 100–150 M uptake rates of NH ₄ ⁺ were 3–4 times higher than those of NO ₃ ^– The preference for NH ₄ ⁺ uptake was also reflected in the minimum concentration (C_min) values. Supplying NH₄NO₃, the rate of NO ₃ ^– uptake was very low until the NH ₄ ⁺ concentrations had fallen below about 100 M. The shift from NH ₄ ⁺ to NO ₃ ^– uptake was correlated with a corresponding shift from net H⁺ production to net H⁺ consumption in the external solution. The uptake rates of NH ₄ ⁺ were correlated with equimolar net production of H⁺. With NO ₃ ^– nutrition net consumption of H⁺ was approximately twice as high as uptake rates of NO ₃ ^– In the forest stand the NO ₃ ^– concentration in the soil solution was more than 10 times higher than the NH ₄ ⁺ concentration (<100 M), and the rhizosphere pH of non-mycorrhizal roots considerably higher than the bulk soil pH. The rhizosphere pH increase was particularly evident in apical root zones where the rates of water and NO ₃ ^– uptake and nitrate reductase activity were also higher. The results are summarized in a model of water and nutrient transport to, and uptake by, non-mycorrhizal roots of Norway spruce in a forest stand. Model calculations indicate that delivery to the roots by mass flow may meet most of the plant demand of nitrogen and calcium, and that non-mycorrhizal root tips have the potential to take up most of the delivered nitrate and calcium.     

Quantitative imaging of rhizosphere pH and CO2 dynamics with planar optodes

Background and Aims

Live imaging methods have become extremely important for the exploration of biological processes. In particular, non-invasive measurement techniques are key to unravelling organism–environment interactions in close-to-natural set-ups, e.g. in the highly heterogeneous and difficult-to-probe environment of plant roots: the rhizosphere. pH and CO₂ concentration are the main drivers of rhizosphere processes. Being able to monitor these parameters at high spatio-temporal resolution is of utmost importance for relevant interpretation of the underlying processes, especially in the complex environment of non-sterile plant–soil systems. This study introduces the application of easy-to-use planar optode systems in different set-ups to quantify plant root–soil interactions.

Methods

pH- and recently developed CO₂-sensors were applied to rhizobox systems to investigate roots with different functional traits, highlighting the potential of these tools. Continuous and highly resolved real-time measurements were made of the pH dynamics around Triticum turgidum durum (durum wheat) roots, Cicer arietinum (chickpea) roots and nodules, and CO₂ dynamics in the rhizosphere of Viminaria juncea.

Key Results

Wheat root tips acidified slightly, while their root hair zone alkalized their rhizosphere by more than 1 pH unit and the effect of irrigation on soil pH could be visualized as well. Chickpea roots and nodules acidified the surrounding soil during N₂ fixation and showed diurnal changes in acidification activity. A growing root of V. juncea exhibited a large zone of influence (mm) on soil CO₂ content and therefore on its biogeochemical surrounding, all contributing to the extreme complexity of the root–soil interactions.

Conclusions

This technique provides a unique tool for future root research applications and overcomes limitations of previous systems by creating quantitative maps without, for example, interpolation and time delays between single data points.     

Forest soil respiration across three climatically distinct chronosequences in Oregon

To assess the relative influence of edaphoclimatic gradients and stand replacing disturbance on the soil respiration of Oregon forests, we measured annual soil respiration at 36 independent forest plots arranged as three replicates of four age classes in each of three climatically distinct forest types. Annual soil respiration for the year 2001 was computed by combining periodic chamber measurements with continuous soil temperature measurements, which were used along with site-specific temperature response curves to interpolate daily soil respiration between dates of direct measurement. Results indicate significant forest type, age, and type × age interaction effects on annual soil respiration. Average annual soil respiration was 1100–1600, 1500–2100, and 500–900 g C m⁻² yr⁻¹ for mesic spruce, montane Douglas-fir, and semi-arid pine forests respectively. Age related trends in annual soil respiration varied between forest types. The variation in annual soil respiration attributable to the climatic differences between forest types was 48%(CV). Once weighted by the age class distribution for each forest type, the variation in annual soil respiration attributable to stand replacing disturbance was 15%(CV). Sensitivity analysis suggests that the regional variation in annual soil respiration is most dependent on summer base rates (i.e. soil respiration normalized to a common temperature) and much less dependent on the site-specific temperature response curves (to which annual rates are relatively insensitive) and soil degree-days (which vary only 10% among plots).     

Water uptake by barley roots as affected by the osmotic and matric potential in the rhizosphere

Summary The water uptake rates of roots in saline soils are depressed by the simultaneously decreasing matric and osmotic water potentials in the soil surrounding the roots (rhizospheric soil). Unfortunately there are no reliable tools available for direct measurements of the effect of decreasing water potentials in the rhizospheric soil on the uptake rate of soil water by roots. This paper presents some results of a vegetation technique for studying the effect of different combinations of osmotic and matric water potentials in the rhizospheric soil on the water uptake rates of barley roots. Water uptake rates were reduced to a greater extent by decreasing soil matric water potentials than by decreasing soil osmotic water potentials. According to the results of this experiment, there was no relationship between the total soil water potential of a sandy soil and the water uptake rates when the roots were exposed to different combinations of and .     

Predicted microbial biomass in the rhizosphere of barley in the field

Summary Laboratory data for the loss of root material by barley and field data for the growth of barley plants in Syria and in England have been combined to predict the amount of material lost by barley roots during a season, and to predict the resulting microbial biomass in the rhizosphere. The predicted microbial biomass C in the rhizosphere ranged from 10–34% of the total plant biomass C depending mainly upon the value used for rate of loss of root material. Total loss of root material predicted during a season in England constituted 7.7–25.4 percent of C fixed by photosynthesis. The major assumptions made in these calculations are considered, and the predicted values discussed in relation to reported values for soil microbial biomass, CO₂ fluxes from soil and associative nitrogen fixation.     

Feedback interactions between needle litter decomposition and rhizosphere activity

The aim of our study was to identify interactions between the decomposition of aboveground litter and rhizosphere activity. The experimental approach combined the placement of labelled litter (¹³C=–37.9) with forest girdling in a 35-year-old Norway spruce stand, resulting in four different treatment combinations: GL (girdled, litter), GNL (girdled, no litter), NGL (not girdled, litter), and NGNL (not girdled, no litter). Monthly sampling of soil CO₂ efflux and ¹³C of soil respired CO₂ between May and October 2002 allowed the partitioning of the flux into that derived from the labelled litter, and that derived from native soil organic matter and roots. The effect of forest girdling on soil CO₂ efflux was detectable from June (girdling took place in April), and resulted in GNL fluxes to be about 50% of NGNL fluxes by late August. The presence of litter resulted in significantly increased fluxes for the first 2 months of the experiment, with significantly greater litter derived fluxes from non-girdled plots and a significant interaction between girdling and litter treatments over the same period. For NGL collars, the additional efflux was found to originate only in part from litter decomposition, but also from the decay of native soil organic matter. In GL collars, this priming effect was not significant, indicating an active role of the rhizosphere in soil priming. The results therefore indicate mutual positive feedbacks between litter decomposition and rhizosphere activity. Soil biological analysis (microbial and fungal biomass) of the organic layers indicated greatest activity below NGL collars, and we suppose that this increase indicates the mechanism of mutual positive feedback between rhizosphere activity and litter decomposition. However, elimination of fresh C input from both above- and belowground (GNL) also resulted in greater fungal abundance than for the NGNL treatment, indicating likely changes in fungal community structure (i.e. a shift from symbiotic to saprotrophic species abundance).     

The effects of low pH on the respiration of Daphnia magna Straus.

The effect of low pH values on Daphnia magna is shown to severely depress the O₂ uptake rates. The reason for this is discussed with respect to a decrease in the ability of the gills to exchange CO₂ with the surrounding medium due to a reduction in the CO₂ diffusion gradient as a result of increasing acidity. Respiration in CO₂ free water at acid pH values is shown to be apparently unaffected.     

Variability in soil respiration across riparian-hillslope transitions

The spatial and temporal controls on soil CO₂ production and surface CO₂ efflux have been identified as outstanding gaps in our understanding of carbon cycling. We investigated both across two riparian-hillslope transitions in a subalpine catchment, northern Rocky Mountains, Montana. Riparian-hillslope transitions provide ideal locations for investigating the controls on soil CO₂ dynamics due to strong, natural gradients in the factors driving respiration, including soil water content (SWC) and soil temperature. We measured soil air CO₂ concentrations (20 and 50 cm), surface CO₂ efflux, soil temperature, and SWC at eight locations. We investigated (1) how soil CO₂ concentrations differed within and between landscape positions; (2) how the timing of peak soil CO₂ concentrations varied across riparian and hillslope zones; and (3) whether higher soil CO₂ concentrations necessarily resulted in higher efflux (i.e. did surface CO₂ efflux follow patterns of subsurface CO₂)? Soil CO₂ concentrations were significantly higher in the riparian zones, likely due to higher SWC. The timing of peak soil CO₂ concentrations also differed between riparian and hillslope zones, with highest hillslope concentrations near peak snowmelt and highest riparian concentrations during the late summer and early fall. Surface CO₂ efflux was relatively homogeneous at monthly timescales as a result of different combinations of soil CO₂ production and transport, which led to equifinality in efflux across the transects. However, efflux was 57% higher in the riparian zones when integrated to cumulative growing season efflux, and suggests higher riparian soil CO₂ production.     

Secretion of momilactone A from rice roots to the rhizosphere

The secretion levels of momilactone A from rice (Oryza sativa L.) seedlings of eight cultivars into the rhizosphere were compared with the endogenous momilactone A concentrations in their shoots and roots. All rice cultivars contained momilactone A in the shoots and roots, and concentrations differed among the cultivars. Momilactone A was also found in all culture solutions in which the rice seedlings were grown, and the concentrations differed among the cultivars. The momilactone A concentrations in the culture solutions were reflected in the momilactone A concentrations in the shoots. These results suggest that all rice cultivars may produce momilactome A and secrete momilactone A into the culture solutions. The secretion levels of momilactone A may be more dependent on their capacities for momilactone A production in the shoots than on their capacities for momilactone A transportation from the shoots into the environment via the roots. As momilactone A acts as an antimicrobial and allelopathic agent, the secretion of momilactone A into the rice rhizosphere may provide a competitive advantage for root establishment through local suppression of soil microorganisms and inhibition of the growth of competing plant species.     

Effect of rhizosphere pH on the availability and uptake of Fe,Mn and Zn

In pot experiments the relationships between rhizosphere pH, the extractable levels of Fe, Mn and Zn in the soil and their uptake into the roots and shoots of dwarf French beans have been studied. Variations in rhizosphere pH were induced by applying three different sources of nitrogencholine phosphate, ammonium phosphate and calcium nitrate-to an initially homogenised soil (preadjusted to either pH 7 or 8). The rhizosphere pH was found to be significantly lower following the application of either ammonium or choline phosphates and to be increased by calcium nitrate treatment.The Fe and Zn contents of both shoot and root were inversely proportional to rhizosphere pH. The Mn contents also increased with decreasing pH but a sharp increase was apparent below pH 5.5. The shoot Fe, Zn and Mn content were significantly correlated with the extractable levels determined in the rhizosphere and non-rhizosphere soil.This paper forms part of the Ph. D. thesis submitted by A.N.S. to the University of Wales in 1977.     

Coupling of canopy gas exchange with root and rhizosphere respiration in a semi-arid forest

The strength of coupling between canopy gas exchange and root respiration was examined in ~15-yr-old ponderosa pine (Pinus ponderosa Doug. Ex Laws.) growing under seasonally drought stressed conditions. By regularly watering part of the root system to reduce tree water stress and measuring soil CO₂ efflux on the dry, distant side of the tree, we were able to determine the strength of the relationship between soil autotrophic (root and rhizosphere) respiration and changes in canopy carbon uptake and water loss by comparison with control trees (no watering). After ~40 days the soil CO₂ efflux rate, relative to pre-treatment conditions, was twice that of the controls. This difference, attributable to root and rhizosphere respiration, was strongly correlated with differences in transpiration rates between treatments (r² = 0.73, p<0.01). By the end of the period, transpiration of the irrigated treatment was twice that of controls. Periodic measurements of photosynthesis under non-light limited conditions paralleled the patterns of transpiration and were systematically higher in the irrigated treatment. We observed no evidence for a greater sensitivity of soil autotrophic respiration to temperature compared to the response of heterotrophic respiration to temperature; the Q₁₀ for total soil respiration was 1.6 (p>0.99) for both treatments. At the ecosystem scale, daily soil CO₂ efflux rate was linearly related to gross primary productivity (GPP) as measured by eddy-covariance technique (r² = 0.55, p<0.01), suggesting patterns of soil CO₂ release appear strongly correlated to recent carbon assimilation in this young pine stand. Collectively the observed relationships suggest some consideration should be given to the inclusion of canopy processes in future models of soil respiration.     

Release of carboxylic anions and protons by tomato roots in response to ammonium nitrate ratio and pH in nutrient solution

The exudation of certain organic anions and protons by roots which may affect solubility of metals and P and uptake by plants, is affected by nitrogen form and pH. The objective of this work was to study exudation of carboxylates and H⁺/OH^– by tomato plants in response to NH₄/NO₃ ratio and pH in nutrient solution. Four NH₄/(NH₄+NO₃) ratios (R= 0, 0.33, 0.67 and 1) and constant vs. variable solution pH treatments were investigated. The sum of the exudation rates of all carboxylates tended to decline with increasing R, particularly tri- and dicarboxylates. The molar fraction of the exuded tri- and dicarboxylates, averaged over all treatments and plant ages, increased in the order tartarate 2%), malate (6%), succinate (15%), citrate (26%) and fumarate (46%). At R=1 the solution pH dropped from 5.2 to 3 and at R=0 increased to 8. The R corresponding to the pH stat of tomato plant was 0.3. For the constant solution pH treatment, the effect of solution pH on carboxylate exudation rate was small as compared to the effect of R. The exudation of citrate and H⁺ efflux which were initiated when NO₃ and NH₄ uptake rates per plant exceeded certain threshold values, increased with plant age.     

Ecology of heterotrophic dinitrogen fixation in the rhizosphere of mangrove plant community at the Ganges river estuary in India

Summary Heterotrophic dinitrogen fixation in root associations of successional stages of the tropical mangrove plant community at the Ganges river estuary in India was investigated by excised-root acetylene reduction assay, and enumeration and identification of diazotrophic bacteria from sediment, root and tidal water samples. High to very high rates of nitrogenase activity (64–130 nmol C₂H₄/g dry root/h) were associated with washed excised roots of seven common early-successional mangrove species at the inundated swamps. Declining, late-successional mangroves at the occasionally inundated ridges had considerably lower values and the “declined” mangroves and other non-littoral species at embankment protected highlands had very low to insignificant values of root nitrogenase activity. Total and inorganic nitrogen contents of the mangrove sediments were low and were positively related to the stages of physiographic succession. Plant-associated sediments of particularly the old formation swamps had very high C/N ratios. Nine isolates of nitrogen-fixing bacteria belonging to all known O₂ response groups were distinguished from a large population of diazotrophs associated with roots of mangroves and other associate plant species of the community. The isolates differed with respect to their N₂-fixation efficiency and halotolerance in pure culture. There was no specificity of any of the bacterial isolates to any of the plant species of the community but a higher number of efficient isolates were seen to be associated with mangroves at the swampy succession. Sediment-free tidal water also contained a large population of microaerophilic and anaerobic N₂-fixing bacteria.     

Effect of elevated atmospheric CO2 concentration on soil and root respiration in winter wheat by using a respiration partitioning chamber

Soil respiration in a cropland is the sum of heterotrophic (mainly microorganisms) and autotrophic (root) respiration. The contribution of both these types to soil respiration needs to be understood to evaluate the effects of environmental change on soil carbon cycling and sequestration. In this paper, the effects of free-air CO₂ enrichment (FACE) on hetero- and autotrophic respiration in a wheat field were differentiated and evaluated by a novel split-root growth and gas collection system. Elevated atmospheric pCO₂ of approximately 200 μmol mol⁻¹ above the ambient pCO₂ significantly increased soil respiration by 15.1 and 14.8% at high nitrogen (HN) and low nitrogen (LN) application rates, respectively. The effect of elevated atmospheric pCO₂ on root respiration was not consistent across the wheat growth stages. Elevated pCO₂ significantly increased and decreased root respiration at the booting-heading stage (middle stage) and the late-filling stage (late stage), respectively, in HN and LN treatments; however, no significant effect was found at the jointing stage (early stage). Thus, the effect of increased pCO₂ on cumulative root respiration for the entire wheat growing season was not significant. Cumulative root respiration accounted for approximately 25–30% of cumulative soil respiration in the entire wheat growing season. Consequently, cumulative microbial respiration (soil respiration minus root respiration) increased by 22.5 and 21.1% due to elevated pCO₂ in HN and LN, respectively. High nitrogen application significantly increased root respiration at the late stage under both elevated pCO₂ and ambient pCO₂; however, no significant effects were found on cumulative soil respiration, root respiration, and microbial respiration. These findings suggest that heterotrophic respiration, which is influenced by increased substrate supplies from the plant to the soil, is the key process to determine C emission from agro-ecosystems with regard to future scenarios of enriched pCO₂.     

Inhibition of the growth of two blue-green algae species (Microsystis aruginosa and Anabaena spiroides) by acidification treatments using carbon dioxide

The effect of pH adjusted by aeration with carbon dioxide (CO₂) on the growth of two species of blue-green algae, Microcystis aeruginosa and Anabaena spiroides, was investigated. Three conditions (pH 5.5, 6.0 and 6.5) were found to have significant inhibitory effects on the growth of the two algae species when acidification treatment was conducted during the logarithmic phase. Differences in the inhibition effect of acidification existed between the two species algae. The tolerance of M. aeruginosa to these conditions was also investigated. The results indicated that M. aeruginosa was inhibited significantly, but not dead at pH 6.5, whereas death occurred at pH 5.5 and 6.0. The greatest inhibitory effect of acidification treatment conducted during the stable breeding phase of M. aeruginosa occurred at pH 5.5, while no inhibitory effect was found at pH 6.5.     

Belowground carbon cycling in a humid tropical forest decreases with fertilization

Only a small fraction of the carbon (C) allocated belowground by trees is retained by soils in long-lived, decay-resistant forms, yet because of the large magnitude of terrestrial primary productivity, even small changes in C allocation or retention can alter terrestrial C storage. The humid tropics exert a disproportionately large influence over terrestrial C storage, but C allocation and belowground retention in these ecosystems remain poorly quantified. Using mass balance and ¹³C isotope methods, we examined the effects of afforestation and fertilization, two land-use changes of large-scale importance, on belowground C cycling at a humid tropical site in Hawaii. Here we report that in unfertilized plots, 80% of the C allocated belowground by trees to roots and mycorrhizae was returned to the atmosphere within 1 year; 9% of the belowground C flux was retained in coarse roots and 11% was retained as new soil C. The gains in new soil C were offset entirely by losses of old soil C. Further, while fertilization early in stand development increased C storage in the litter layer and in coarse roots, it reduced by 22% the flux of C moving through roots and mycorrhizae into mineral soils. Because soil C formation rates related strongly to rhizosphere C flux, fertilization may reduce an already limited capacity of these forests to sequester decay-resistant soil C.     

Establishment of inoculatedAzospirillum spp. in the rhizosphere and in roots of field grown wheat and sorghum

Summary Four field experiments were carried out with wheat or sorghum in different regions of Brazil. The aim was to study the establishment of inoculatedAzospirillum strains, marked with resistance to various antibiotics, in the rhizosphere and in roots. The levels of the various antibiotics were chosen according to the resistance of the indigenousAzospirillum population.Azospirillum brasilense strains Sp 107 and Sp 245 could be established in all three wheat experiments and predominated within theAzospirillum population in washed, and especially in surface sterilized, roots. Strains Sp 7 and Cd established poorly in wheat roots.Azospirillum lipoferum Sp S82 represented 72% of the root isolates from sorghum inoculated with this strain. This strain and naturalAzospirillum infection became concentrated in the upper parts of the root system. Improved methods for root surface sterilization in which the absence ofAzospirillum on the root surface was established by pre-incubating roots with paraffin-capped ends in NFb medium confirmed the establishment of inoculatedAzospirillum strains within sorghum roots in the field.     

Changes in pollen tube behaviour induced by carbon dioxide and their role in overcoming self-incompatibility in Brassica

Summary Comparative studies on self-pollination after the application of high concentrations of CO₂ gas, which is known to overcome the self-incompatibility reaction in Cruciferous species, have revealed significant changes in the pollen tubes of germinating grains prior to their penetration into the papilla cells. A remarkable increase in the width of the pollen tube was induced by treating the stigmatic papillae with high CO₂ concentrations. The width of the pollen tube appeared to be greatest with CO₂ concentrations ranging from 3% to 5%; these concentrations were also optimal for tube penetration. Callose accumulation was extensively induced in the stigmatic papilla with 10%–20% of CO₂, although a typical callosic reaction remained through the ranges appropriate for blocking self-incompatibility. Observations using the scanning electron microscope (SEM) after pollination revealed that compatible pollen tubes in cross-pollinations fused completely to the papular surface during tube penetration, while in self-pollination, pollen tubes remained on the papilla with some additional diffusate. In the case of CO₂ treatment for self-pollination, some pollen tubes behaved very similarly to the incompatible or compatible ones already described, while others were different from both of them: they showed a complete fusion, similar to compatible ones, with additional diffusate, similar to incompatible ones. These responses of the pollen and stigma to high CO₂ concentrations are discussed with respect to their effect upon the expression of self-incompatibility.