Growth and root nodule nitrogenase activity of Pisum sativum as influenced by transpiration

The influence of shoot transpiration on the rates of growth and nitrogen fixation was investigated in Pisum sativum L. cv. Rondo. In short term experiments, rates of transpiration and acetylene reduction of intact plants were measured simultaneously, using air-tight perspex vessels enclosing the basal part of the nodulated root. In long term experiments, accumulation of dry matter and reduced nitrogen in the plant were determined as well. Transpiration rate changed diurnally and was varied by manipulating the vapour saturation deficit or the flow rate of the air in the growth cabinet. The rate of acetylene reduction declined after subjecting intact plants to high transpiration rates. This decline was accompanied by a desiccation of the root nodules. Dry matter and reduced nitrogen accumulation were not affected by transpiration rate. At low transpiration rate reduced nitrogen content of the root nodules was higher than at high transpiration rate. However, in these nodules the rate of acetylene reduction was not significantly affected. It is concluded that the nitrogenase activity of pea root nodules is insensitive to changes in the flow rate and the organic N concentration of the xylem sap within a wide range of transpiration conditions under the applied growth conditions.     

Carbon and nitrogen metabolism in legume root nodules

The literature concerning the metabolism of carbon compounds during the reduction, assimilation and translocation of nitrogen in root nodules of leguminous plants is reviewed. The reduction of dinitrogen requires an energy source (ATP) and a reluctant which are both supplied by respiratory catabolism of carbohydrates produced by the host plant. Photosynthates are also required to generate the carbon skeletons for amino acid or urcide synthesis during the assimilation of ammonia produced by the bacteria within the nodule tissue. Competition for photosynthates occurs between the bacteroids, nodule tissue and the various vegetative and reproductive sinks in the host plant. The nature of carbon compounds involved in these processes, their routes of metabolism, the mechanisms of control and the partitioning of metabolises between the various sites of utilization are only poorly understood. It is apparent that dinitrogen is reduced to ammonia in the bacteroids. Both fast- and slow-growing strains of Rhizobium possess the Entner-Doudoroff pathway of glucose catabolism, and some, if not all, enzymes of the Emden-Meyerhof pathway. Some bacterial cultures also metabolize carbon through the ketogluconate pathway but only the fast-growing strains of cultured rhizobia possess the key enzyme of the pentose phosphate pathway (6-phosphogluconate dehydrogenase). The host cells are thought to contain the complete Emden-Meyerhof pathway and tricarboxylic acid cycle, which provides the carbon skeletons for assimilation of the ammonia, formed by the bacteroids, into α-amino acids. A pathway of anapleurotic carbon conservation, operative in the host cells, synthesizes oxaloacetic acid through β-carboxylation of phosphoenol pyruvate. This process could be important in the recapture and assimilation of respired CO₂ in the rhizosphere. The main route of assimilation of ammonia produced by the bacteroids would appear to be via the glutamine synthetase-glutamate synthase pathway in the host cells. However, glutamate dehydrogenase may also be involved in ammonia assimilation. These enzymes also occur in in vitro cultures of Rhizobium and in bacteroids where they presumably participate in the synthesis of amino acids for growth of the bacteria or bacteroids. Nitrogen assimilated into glutamine or glutamate is exported from the nodules in a variety of forms, which include asparagine, glutamine, aspartate, homoserine and allantoates, in proportions which depend on the legume species. Studies on regulation of the overall process have focussed on expression of bacteroid genes and on the control of enzyme activity, at the level of nitrogenase and enzymes of nitrogen assimilation in particular. However, due to the wide range of experimental techniques, environmental conditions and plant species which have been used, no clear conclusions can yet be drawn. The pathways of carbon flow in nitrogen metabolism, particularly in relation to the synthesis of ureides and the regulation of carbon metabolism, remain key areas for future research in symbiotic nitrogen fixation.     

Nitrogen-fixing root nodules inDatisca Cannabina L.

Summary The occurrence of root nodules inDatisca cannabina is reported. The nodules are typically of the Alnus type, forming dichotomously-branched coralloid clusters. The enlarged cortical cells contain vesicle clusters of the endophyte. The nodules reduced acetylene to ethylene at a rate of 5.5 moles ethylene per g fresh wt of nodules per hour.     

Proteoid root morphology and function inLupinus albus

Summary Current theories of phosphorus uptake by plants imply that they can augment diffusion to their root axes by the development of abundant root hairs or mycorrhizas. Some phosphorus efficient plants have root morphology with multi-branched roots and localised regions of densely packed root hairs, which we suggest is better suited to the retention of substances exuded by the roots than uptake of substances moving to the root by diffusion. Evidence of substantial exudation by the proteoid roots ofLupinus albus is presented.     

In-vitro-cultured subclover root can develop Fe-deficiency stress response

The Fe-deficiency stress response is induced in most plants under Fe-deficient conditions, but whether the shoot and/or the root control development of the stress response is not known. The objectives of the present study were to determine whether in-vitro-cultured subclover roots can develop Fe-deficiency stress response and to examine this approach as a possible screening technique for Fe-deficiency resistance. One-cm long root tips of subclover seedlings were cultured in modified White's medium without (-Fe) or with (+Fe) 100 μM Fe³⁺EDTA. Root Fe³⁺ reduction and H⁺ release were evaluated. On the first day after transfer to the -Fe medium, the Fe-deficiency-resistant cultivar Koala (Trifolium brachycalycinum Katzn. and Morley) started to release H⁺, resulting in a decrease in pH of the culture medium, while the susceptible cultivar Karridale (T. subterraneum L.) did not release H⁺ until the second day. The H⁺-release rate of the -Fe Koala was approximately twice as high as that of the -Fe Karridale for the first 4 days of -Fe treatment. Both Koala and Karridale reached their highest H⁺-release rates on the fourth day after -Fe treatment initiation. The +Fe Koala released H⁺ after several days of culture, but the H⁺ release of the -Fe Koala was severalfold greater than that of the +Fe Koala. The implicit correlation between H⁺ release and Fe-deficiency resistance was substantiated by using a series of subclover cultivars with a range of susceptibilities to Fe deficiency. The pH of the -Fe culture media of the series of cultivars was positively correlated to their Fe-chlorosis scores reported in previous research. The results of the present study indicate that root itself has the full ability to develop Fe-deficiency stress response and the response is dependent on the root Fe status. The results also suggest that root culture could be used as a simple and efficient alternative technique for screening germplasm for Fe-deficiency resistance.     

Reduction of root iron in Plantago lanceolata during recovery from Fe deficiency

Changes in root-associated Fe(III) reductase activity and Fe concentration during recovery from temporary iron starvation were investigated in hydroponically grown Plantago lanceolata L. Within two days, interruption of the Fe supply resulted in enhanced rates of reduction by intact plant roots. Transfer of iron deficient plants to a solution containing various amounts of chelated Fe caused a transient increase in reduction activity before the rates declined to a level determined by the amount of Fe added. Repression of root-associated redox activity was independent of the Fe concentration in the preculture. When iron deficient plants were submitted to a supply of Fe localized to a part of the root system (split-root plants), the decrease in reduction rates was much more pronounced in the Fe-deprived portion of the roots than in the Fe-supplied one. No correlation was observed between root Fe concentration and Fe(III) reductase activity. Continued growth of split-root plants in the +Fe/-Fe regime increased the reduction rates of the +Fe-grown portion of the root system over the rates in iron sufficient plants with non-divided roots. The results are discussed in relation to putative factors mediating intra- and interorgan regulation of iron nutrition.     

Reduction of root iron in Plantago lanceolata during recovery from Fe deficiency

Changes in root-associated Fe(III) reductase activity and Fe concentration during recovery from temporary iron starvation were investigated in hydroponically grown Plantago lanceolata L. Within two days, interruption of the Fe supply resulted in enhanced rates of reduction by intact plant roots. Transfer of iron deficient plants to a solution containing various amounts of chelated Fe caused a transient increase in reduction activity before the rates declined to a level determined by the amount of Fe added. Repression of root-associated redox activity was independent of the Fe concentration in the preculture. When iron deficient plants were submitted to a supply of Fe localized to a part of the root system (split-root plants), the decrease in reduction rates was much more pronounced in the Fe-deprived portion of the roots than in the Fe-supplied one. No correlation was observed between root Fe concentration and Fe(III) reductase activity. Continued growth of split-root plants in the +Fe/-Fe regime increased the reduction rates of the +Fe-grown portion of the root system over the rates in iron sufficient plants with non-divided roots. The results are discussed in relation to putative factors mediating intra- and interorgan regulation of iron nutrition.     

根系活动对湿地植物根际铁异化还原的影响及机制研究进展

根系活动是影响湿地植物根际铁异化还原速率的关键因素之一。以往国内外湿地铁异化还原的研究多为分析和比较各类中宏观生境中铁异化还原能力的差异。近年来,湿地植物根际微域铁的生物地球化学行为也日益成为该领域的研究热点。综述了根际铁异化还原研究概况,梳理了根系活动对根际铁异化还原关键因子的作用机制,分析了根际铁异化还原和其他有机质代谢途径的竞争关系,探讨了根际铁异化还原对根系活动动态变化和异质性的响应,提出了根际铁异化还原的概念模型,并指出了未来我国湿地植物根际铁异化还原研究应加强的工作。     

The acetylene-induced decline in nitrogenase activity in root nodules of Elaeagnus angustifolia

The rate of C₂H₂ reduction by nodulated seedlings of Elaeagnus angustifolia (Russian olive) was followed as a function of time. Our goals were to: 1) determine whether there is an C₂H₂-induced decline in nitrogenase activity; and 2) investigate the mechanism of any decline. We found a peak rate of C₂H₂ reduction at 1.5 min after the introduction of C₂H₂ that was followed by a rapid decline in activity to 56% of the peak value. After the decline there was a partial recovery to 67% of the peak value at 60 min. When the pO₂ was decreased during the decline there was no significant effect (p0.05) on nitrogenase activity. When the C₂H₂ reduction assay was preceded by an incubation in a gas mixture (20 kPa O₂) with Ar substituted for N₂, there was little decline in nitrogenase activity as a function of time, but the rate of C₂H₂ reduction per gram nodule was reduced by approximately 50%. From these results we conclude that t Elaeagnus angustifolia exhibits a pronounced C₂H₂-induced decline and consequently the initial peak rate C₂H₂ reduction must be determined to obtain a valid measure of nitrogenase activity. We further suggest that cessation of NH₃ formation initiates the decline and that the decline is not caused by a change in nodule permeability to gases.     

The acetylene reduction assay inactivates root nodule uptake hydrogenase in some actinorhizal plants

Actinorhizal nodules do not usually evolve H₂ due to the action of an uptake hydrogenase. We have found that nodules of several Frankia symbioses evolved large amounts of H₂ gas when returned to air following exposure to 10 kPa C₂HT₂ during an acetylene reduction assay. Increased H₂ evolution in air persisted for several days when intact root systems of Alnus incana (L.) Moench (inoculated with Frankia UGL 011101) were treated with 10 kPa C.H₂ for 1 h. Full recovery of uptake hydrogenase activity required 4 to 8 days. Studies with crude homogenates of nodules of the same plants showed that hydrogenase (measured amperometrically with phenazine metho-sulfate as electron acceptor) was directly affected, since activity in treated nodules was only 10% of that in untreated nodules. A survey of actinorhizal symbioses revealed variation in the effect of an acetylene reduction assay on hydrogen metabolism. Nodules of three species, including Alnus rubra Bong, inoculated with Frankia HFPArD. showed complete inactivation of hydrogenase. H₂ evolution in air was 25% of the C₂H₂ reduction rate and H, evolution in Ar/O₂ was equal to the QH₂ reduction rate. Two symbioses, Ceanothus americanus L. (soil inoculant) and Batista glomerata Baill. (soil inoculant) showed no change following an acetylene reduction assay. A third group of symbioses showed an intermediate response.     

Ethylene production,cluster root formation,and localization of iron(III) reducing capacity in Fe deficient squash roots

Dicots and non-graminaceous monocots have the ability to increase root iron(III) reducing capacity in response to iron (Fe) deficiency stress. In squash (Cucurbita pepo L.) seedlings, Fe(III) reducing capacity was quantified during early vegetative growth. When plants were grown in Fe-free solution, the Fe(III) reducing capacity was greatly elevated, reached peak activity on day 4, then declined through day 6. Root ethylene production exhibited a temporal pattern that closely matched that of Fe(III) reducing capacity through day 6. On the 7th day of Fe deficiency, cluster root morphology developed, which coincided with a sharp increase in the root Fe(III) reducing capacity, although ethylene production decreased. Localization of Fe(III) reducing capacity activity was observed during the onset of Fe deficiency and through the development of the root clusters. It was noted that localization shifted from an initial pattern which occurred along the main and primary lateral root axes, excluding the apex, to a final localization pattern in which the reductase appeared only on secondary laterals and cluster rootlets. This revised version was published online in June 2006 with corrections to the Cover Date.     

Injection of hydrogen gas stimulates acid mine drainage treatment in laboratory‐scale hydroponic root mats

The environmentally benign disposal of acid mine drainage (AMD) is still a technical challenge. In the present study, artificial AMD was treated in a laboratory‐scale floating hydroponic root mat of soft rush, Juncus effusus. This ecotechnological system was operated with hydrogen injection and water recirculation but without an external carbon supply. It achieved a mean increase of ΔpH = 3.3 up to pH ≈ 8.2, high sulfate removal of up to 87%, and efficient removal of iron (100%), aluminum (99.8%), manganese (97.4%), and zinc (99.6%). Sulfide was not detected in the outflow. Treatment performance correlated with the amount of hydrogen loading. Daily oscillations of the redox potential up to amplitudes of ΔE_h ≈ 450 mV in a mean range of E_h ≈ ?150 to +300 mV indicated a correlation of plant physiology and removal processes. Apparently, sulfate and metal removal were the result of chemolithotrophic microbial sulfate reduction supported by the externally provided H₂ and chemoorganotrophic sulfate reduction driven by rhizodeposits. Bicarbonate generated in the microbial transformation of such plant‐derived organic carbon contributed to pH neutralization. The effluent's pH increase was governed further by recirculation of the treated AMD. The flow regime and the injection of hydrogen at the ground of the root mat caused concentration gradients where the most efficient removal occurred in the deepest zone of the root mat. Further investigations should target long‐term stability, plant growth dynamics, load variations, balances of carbon and sulfur, the removal of H₂S and metal precipitates from the system as well as efficient hydrogen supply.     

Measurement of nitrogenase activity in legume root nodules: In defense of the acetylene reduction assay

The closed acetylene reduction assay has been used as a measure of nitrogenase activity and an indicator of N₂ fixation in Rhizobium/legume symbioses for 25 years. However, starting 10 years ago this assay has come under harsh criticism as being inaccurate. Currently, confusion exists regarding the conditions under which the acetylene reduction assay can be used accurately, or whether it can be used at all as a measure of nitrogenase activity. This article reviews the circumstance that has lead to this confusion. The author argues that under the proper assay conditions and with the appropriate checks, the closed acetylene reduction assay is still a valuable tool in assessing relative differences in nitrogenase activity in Rhizobium/legume symbioses.     

Ruzigrass root persistence and soybean root growth

Plant and Soil - Cropping systems using forage grasses as cover crops have been effective in soil conservation and nutrient cycling, but root persistence of ruzigrass (Urochloa ruziziensis) is...     

Wheat root diversity and root functional characterization

Background and Aims

Under limited moisture conditions, roots can play an outstanding role with respect to yield stability by effective absorption of water from soil. A targeted integration of root traits into plant breeding programs requires knowledge on the existing root diversity and access to easy and cost-effective methods. This study aimed to assess wheat root diversity, root properties in relation to water regime, and the efficiency of root capacitance for in situ screening.

Methods

Root morphological, anatomical properties and root capacitance of wheat species from different ploidy levels were studied under field conditions in 2 years contrasting in water regime. Soil water content was weekly measured.

Results

Significant genotypic differences were observed for most root traits. The investigated genotypes exploited different strategies to maximize soil water depletion, e.g. high topsoil root length density, low tissue mass density, high specific root length, deep rooting and looser xylem vessels. Multivariate statistics of root traits revealed an acceptable genotypic differentiation according to regional origin, genetics and capacity to extract soil water.

Conclusions

Under supply-driven environments, dehydration avoidance via water uptake maximization can be achieved through high topsoil rooting density. In this regard, root capacitance can be useful for in situ screening.     

Effects of european alder (Alnus flutinosa (L.) Gaertn) rhizobacteria on nodular metabolism and root development

The effects of 3 Bacillus and 7 Pseudomonas strains on development of the root system and nodular metabolism, evaluating CO₂ production and acetylene reduction activity (ARA), of Alnus glutinosa, were studied. All experiments were done on nodulated plants (N) with the symbiont Frankia and on non-nodulated plants (NN).An increase in root length (RL) and root surface (RS) was detected when growth culture media from three different Bacillus free of bacteria were assayed, both in N and NN plants. However, Pseudomonas growth culture media reduced RS in N plants, and a decrease in RL parallel to an increase in RS in NN plants. Bacillus growth cultyre media caused an increase: and CO₂ production while Pseudomonas culture media caused lower ARA and a noticeable increase in nodular respiration. Results are discussed considering nutritional and/or hormonal (Bacillus) or phytotoxic factors (Pseudomonas).     

Stereoselective reduction of ketones by <Emphasis Type="Italic">Daucus carota</Emphasis> hairy root cultures

Reduction of acetophenone by Daucus carota hairy root cultures afforded (S)-phenylethanol in high yield (96%) and excellent enantiomeric excess (ee ≥ 98%). Aromatic ketones, keto esters, and a simple aliphatic ketone were reduced with good stereoselectivity (ee = 62–98%) and moderate to high chemical yields (25–90%).     

Effects of age, supra-ambient oxygen and repeated assays on acetylene reduction and root respiration in pea

An open flow-through gas system was used to investigate the effect of plant age on nitrogenase activity in relation to root respiration (measured as CO₂ release) and supra-ambient O₂ levels in 24- to 51-day-old, nodulated Pisum sativum L. cv. Bodil. The effect of assaying plants repeatedly was also studied. The respiratory efficiency of nitrogenase [mol CO₂ (mol C₂H₄)⁻¹] and the relative decline in nitrogenase (EC 1.7.99.2) activity in response to introduction of C₂H₂ in the gas stream were unaffected by plant age. In contrast, the nitrogenase-linked respiration as a proportion of total root respiration increased with time. Accordingly, the specific respiration linked-to growth and maintenace of the noduled root system decreased with time. C₂H₂ reduction and root respiration were increased by supra-ambient O₂ levels, but the tolerance to high O₂ concentrations seemed to decrease with plant age. Repeated C₂H₂ assays on the same plants decreased their rate of growth and N accumulation: in addition, nitrogenase activity and root respiration were somewhat negatively affected. The results indicate that results from experiments with plants of different ages cannot always be directly compared, and that repeated C₂H₂ assays on the same plants should be applied with caution in physiological work.