Ontogenetic Changes in Root Nodule Subpopulations of Common Bean (Phaseolus vulgaris L.): II. PROTEIN CONTENT AND CARBOHYDRATE POOLS

Root nodules of common bean (Phaseolus vulgaris L. GN 1140)show different ontogenetic patterns of nitrogen fixation indifferent parts of the root system. The objective of the presentstudy was to search for nodule physiological factors which couldexplain these differences. Plants were grown to maturity undercontrolled environment conditions. Root nodules were regularlyharvested from two zones of the root system, and assayed forsoluble cytosolic content and carbohydrate status. Nodule concentrationsof soluble carbohydrates, glucose, fructose or sucrose, didnot change significantly during the pod-filling decline in nitrogenaseactivity. Nodule concentrations of sucrose appeared more relatedto nodule ontogeny than to the ontogenetic stage of the shoot.Total soluble protein and leghaemoglobin concentrations werehighly correlated to nitrogenase activity. In the top zone ofthe root system proteolytic capacity increased during pod-filling,indicating a role in nodule senescence. Proteolytic capacityalso increased in the mid zone, but to a lesser extent, andwithout correlation to nitrogenase activity. The two nodulesubpopulations showed different nitrogenase activity in relationto their leghaemoglobin and carbohydrate status. We suggestthat physiological differences within the root nodule subpopulations,possibly in combination with the distance from the shoot, makesthe top zone more susceptible to a systemic increase in proteolyticcapacity during rapid pod-filling. Key words: Nitrogen fixation, protein, carbohydrate, ontogeny, Phaseolus vulgaris     

Ontogenetic Changes in Root Nodule Subpopulations of Common Bean (Phaseolus vulgaris L.): I. NITROGENASE ACTIVITY AND RESPIRATION DURING POD-FILLING

Nitrogenase activity is commonly measured on a whole plant basis,or only on parts of the root system. In the present paper, activityin different root nodule subpopulations was followed throughoutreproductive growth, in order to characterize the pod-fillingdecline in nitrogenase activity. Inoculated common bean plantswere grown to maturity under controlled environment conditions.Nitrogenase activity (H₂ evolution in air) and nodule respiration(CO₂ evolution) were measured in three separate zones of theroot system with a non-destructive, open flow, gas-exchangesystem. Nitrogenase activity in the top zone drastically declinedat the initiation of pod-filling, whereas nitrogenase activityin the mid zone was stable during the same period. Hence, thepod-filling decline was limited to a certain nodule subpopulationand not of a systemic type. Nodule respiration showed a similar,but less pronounced pattern. The sharp decline in nitrogenaseactivity was not paralleled in nitrogenase specific activity.Nitrogenase activity is not likely to be limited by the availabilityof oxygen or carbohydrates at the onset of pod-filling becausespecific nodule respiration did not change significantly atthis time. In the top zone, nitrogenase specific activity declinedgradually throughout the measurements, whereas in the mid-partof the root system specific activity peaked and gradually declined2-4 weeks later. The dissimilarities between specific and totalnitrogenase activity were explained by differences in nodulegrowth rates. The data suggest that the oldest nodule populationloses activity at the onset of pod-filling. At the same time,nodules grow and nitrogenase activity increases in younger distalparts of the root system. Estimating total nitrogen fixationin this symbiosis by partial sampling of nodulated root systemsis likely to be very misleading. Key words: Nitrogen fixation, respiration, pod-filling decline, Phaseolus vulgaris, ontogeny     

Gas-exchange activity,carbohydrate status,and protein turnover in root nodule subpopulations of field pea (Pisum sativum L. cv. Century)

Root nodule ontogeny was followed in different parts of the root system of field peas (Pisum sativum L. cv. Century) to investigate the contribution to total nitrogen fixation by different nodule subpopulations. Seed-inoculated plants were grown to maturity in controlled-environment growth chambers. In a flow-through system nitrogenase activity (H₂-evolution in air) and nodulated-root respiration (net CO₂-evolution) were measured weekly or biweekly in different parts (top and mid) of the root system. Root nodule extracts were assayed for total soluble cytosolic protein, total heme, proteolytic capacity (at pH 7.0), soluble carbohydrates and starch. Total nitrogenase activity and nodule respiration were higher in the top zone, which was explained by differences in root and nodule mass. Nodule specific nitrogenase activity was similar in both zones, and gradually declined throughout the experiment. No differences were found between nodule subpopulations in the dry-matter specific concentrations of glucose, fructose, sucrose or starch. Neither did nodule concentrations of protein or leghemoglobin differ between the zones. Throughout reproductive growth, no decline was found in total or nodule specific nitrogenase activity, in any of the nodule subpopulations. Growth of the root nodules continued throughout the experiment, though growth of shoot and roots had ceased. The data gives no support for carbohydrate limitation in root nodules during pod-filling, since nodule respiration remained high, the concentration of soluble carbohydrates increased significantly, and the amount of starch was not reduced. We conclude that when this symbiosis is grown under controlled conditions, nitrogenase activity in nodules sub-sampled from the crown part of the root system is representative for the whole nodule population.     

Carbon cost of nitrogenase activity in Frankia–Alnus incana root nodules

The carbon cost of nitrogenase activity was investigated to determine symbiotic efficiency of the actinorhizal root nodule symbiosis between the woody perennial Alnus incana and the soil bacterium Frankia. Respiration (CO₂ production) and nitrogenase activity (H₂ production) by intact nodulated root systems were continuously recorded in short-term assays in an open-flow gas exchange system. The assays were conducted in N₂:O₂, thus under N₂-fixing conditions, in all experiments except for one. This avoided the declines in nitrogenase activity and respiration due to N₂ deprivation that occur in acetylene reduction assays and during extended Ar:O₂ exposures in H₂ assays. Two approaches were used: (i) direct estimation of root and nodule respiration by removing nodules, and (ii) decreasing the partial pressure of O₂ from 21 to 15% to use the strong relationship between respiration and nitrogenase activity to calculate CO₂/H₂. The electron allocation of nitrogenase was determined to be 0.6 and used to convert the results into moles of CO₂ produced per 2e^– transferred by nitrogenase to reduction of N₂. The results ranged from 2.6 to 3.4mol CO₂ produced per 2e^–. Carbon cost expressed as gC produced per gN reduced ranged from 4.5 to 5.8. The result for this actinorhizal tree symbiosis is in the low range of estimates for N₂-fixing actinorhizal symbioses and crop legumes. Methodology and comparisons of root nodule physiology among actinorhizal and legume plants are discussed.     

Inoculationin vitro of wheat seedlings and wheat straw cultures withBacillus azotofixans

Bacillus azotofixans is a recently described species capable of fixing molecular nitrogen efficiently.Ecological studies performed in monoxenic wheat cultures, both in 0.7% agar and in vermiculite-sand mixture, showed that no acetylene reduction occurred and that this bacteria did not grow when supplied only with the wheat plant root exudates. However, after glucose addition to the 0.7% agar cultures, acetylene reduction ability (ARA) was detected. Comparing ARA for media with glucose both with and without plants, it was observed that the plants supply some component leading to the increase of the nitrogenase activity, since the ARA doubled in the samples containing plants.In wheat straw cultures a fast growth of the bacteria was observed in the first 24 hours after inoculation, but no acetylene reduction was detected. After glucose addition to the media with and without straw, nitrogenase activity was detected.     

Nitrogenase — hydrogenase relationships in Rhizobium japonicum

The conditions necessary for coordinate derepression of nitrogenase and O₂-dependent hydrogenase activities in free-living cultures of Rhizobium japonicum were studied. Carbon sources were screened for their ability to support nitrogenase, and then hydrogenase activities. There was a positive correlation between the level of nitrogenase and corresponding hydrogenase activities among the various carbon substrates. The carbon substrate -ketoglutarate was able to support the highest levels of both nitrogenase and hydrogenase activities. When cells were incubated in -ketoglutarate-containing medium, without added H₂ but in the presence of acetylene (to block H₂ evolution from nitrogenase) significant hydrogenase activity was still observed. Complete inhibition of nitrogenase-dependent H₂ evolution by acetylene was verified by the use of a Hup^- mutant. Hydrogen is therefore not required to induce hydrogenase. The presence of 10% acetylene inhibited derepression of hydrogenase. Constitutive (Hup^c) mutants were isolated which contained up to 9 times the level of hydrogenase acitivity than the wild type in nitrogenase induction medium. These mutants did not have greater nitrogenase activities than the wild type.This is contribution number 1254 from the Department of Biology and the McCollum-Pratt Institute Abbreviations: -Ketoglutarate-containing medium (LOKG) and pre-adaptation medium (SRM) as described in Materials and methods     

Analysis of factors affectingin situ nitrogenase (C2H2) activity ofGalega orientalis,Trifolium pratense andMedicago sativa in temperate conditions

Summary A practical fibreglass cylinder-plastic bag system has been designed for making acetylene reduction assays in the field. Thein situ assay was used to determine seasonal patterns of nitrogenase activity for the perennial forage legumesGalega orientalis, Trifolium pratense andMedicago sativa grown under stadard management in southern Filand (60° north). Nitrogenase activity was still detected in the field plots in November, when soil temperature was 1.5°C and air temperature 0.5°C. The acetylene reduction data from weekly measurements were analyzed for correlation with plant growth rate and short-term fluctuations of environmental factors. Generally, there was a good correlation between nitrogenase activity and plant growth rate. Residual fluctuations in activity were only correlated with environmental factors in one case. The nitrogenase activity ofM. sativa was dependent on air temperature in addition to growth rate. Thus, the nitrogen fixing systems in these forage legumes seem to be an integrated part of the plants, being fairly insensitive to short-term environmental changes.Dedicated to Prof. Helge Gyllenberg on the occasion of his 60th birthday.     

Utilization of carbon and nitrogen reserves of alfalfa roots in supporting N2-fixation and shoot regrowth

Perennial legume such as alfalfa have the capacity to sustain shoot regrowth and some nodule N₂-fixation after removal (cutting) of shoots which contain practically all of the plant's photosynthetic capacity. The role of the roots in supporting these processes has not been fully described. Measurements were made of the nodules' responses to removal of shoots from 8-week-old seedlings in terms of N₂-fixation, as nitrogenase activity (NA) measured as acetylene reduction, dark CO₂ fixation, measured as in vitro phosphoenolpyruvate carboxylase (PEPC) activity, and total non-structural carbohydrate (NSC) content. These properties decreased and recovered in that sequence, which suggests that nodule NSC supported the substrate requirements of NA and PEPC immediately after cutting. The utilization and redistribution or root carbon and nitrogen, prelabeled with ¹⁴C and ¹⁵N, were also followed after cutting 8-week-old alfalfa seedlings. In the first 2 weeks of regrowth 12% of root C and 25% of root N were transferred for incorporation into new shoots. Up to 40% of the root C was used for plant respiration to support 28 days of shoot regrowth and N₂-fixation. The decline of N₂-fixation was slower after cutting and its minimum activity rose up 45% of pre-cut activity as root reserves were built up with plant age. Therefore, the stored reserves of nodulated roots play an important role in support of N₂-fixation after cutting.Contribution No. 1265 from Plant Research Center.Contribution No. 1265 from Plant Research Center.     

Inheritance of resistance to potyviruses in Phaseolus vulgaris L. II. Linkage relations and utility of a dominant gene for lethal systemic necrosis to soybean mosaic virus

Summary A single dominant factor, Hss, that conditions a rapid lethal necrotic response to soybean mosaic virus (SMV) has been identified in Phaseolus vulgaris L. cv. Black Turtle Soup, line BT-1. Inoculated plants carrying this factor developed pinpoint necrotic lesions on inoculated tissue followed by systemic vascular necrosis and plant death within about 7 days, regardless of ambient temperature. BT-1 also carries dominant resistance to potyviruses attributed to the tightly linked or identical factors, I, Bcm, Cam, and Hsw, so linkage with Hss was evaluated. No recombinants were identified among 381 F₃ families segregating for potyvirus susceptibility, thus if Hss is a distinct factor, it is tightly linked to I, Bcm, Cam, and Hsw. BT-1 was also crossed reciprocally with the line Great Northern 1140 (GN 1140) in which the dominant gene, Smv, for systemic resistance to SMV was first identified. Smv and Hss segregated independently and are co-dominant. The (GN 1140 x BT-1) F₁ populations showed a seasonal shift of the codominant phenotype. Evaluation of the (GN 1140 x BT-1) F₂ population under conditions where Smv is partially dominant allowed additional phenotypic classes to be distinguished. Pathotype specificity has not been demonstrated for either Smv or Hss. Genotypes that are homozygous for both dominant alleles are systemically resistant to the virus and in addition show undetectable local viral replication or and no seed transmission. This work demonstrates that a gene which conditions a systemic lethal response to a pathogen may be combined with additional gene(s) to create an improved resistant phenotype.     

Growth kinetics and nitrogenase induction inFrankia sp. HFPArI 3 grown in batch culture

Summary Kinetics of growth and nitrogenase induction inFrankia sp. Ar13 were studied in batch culture. Growth on defined medium with NH ₄ ⁺ as the N source displayed typical batch culture kinetics; however, a short stationary phase was followed by autolysis. Removal of NH ₄ ⁺ arrested growth and initiated vesicle differentiation. Vesicle numbers increased linearly and were paralleled by a rise in nitrogenase (acetylene reduction) activity. Nitrogenase activity (10 nM C₂H₄·mg protein^–1·min^–1) was sufficient to support growth on N₂ and protein levels rose in parallel with nitrogenase induction. Optimal conditions for vesicle and nitrogenase induction were investigated. Maximum rates of acetylene reduction were obtained with 5 to 10 mM K₂ HPO₄/KH₂PO₄, 0.1 mM CaCl₂ and MgSO₄. The optimum pH for acetylene reduction and respiration was around 6.7. The amount (5 to 10 g protein/ml) and stage (exponential) of growth of the ammonium-grown inoculum strongly influenced the subsequent development of nitrogenase activity. Propionate was the most effective carbon source tested for nitrogenase induction. Respiration in propionate-grown cells was stimulated by CO₂ and biotin, suggesting that propionate is metabolized via the propionyl CoA pathway.     

Some characteristics of symbiotic nitrogen fixation,yield, protein and oil accumulation in irrigated peanuts (Arachis hypogaea L.)

Summary The potential of peanuts for symbiotic nitrogen fixation is considerable and under optimal edaphic and climatic conditions it reached 222 kg N₂/ha, which was 58% of the nitrogen accumulated in the plants. The effect of the Rhizobium inoculation on crude protein accumulation in the yield (kg/ha) was 3–4 times greater than its effect on the yield of pods and hay. There was an inverse relationship between the protein and oil content in the kernels.Seasonal changes in nitrogenase activity in the nodules were determined by the acetylene reduction method during two growing seasons. Under favorable conditions the specific activity of the nitrogenase reached a very high level (up to 975 moles C₂H₂ g dry wt nod/h) and the total activity (moles C₂H₄/plant/h) was also high in spite of the relatively poor nodulation (weight and number). The high activity was drastically reduced (to 75 moles C₂H₄ g dry wt nod/h) due to exceptionally hot and dry weather, which occurred in the middle of the second half of the growing season. It appears that N₂-fixation (nitrogenase activity) is more sensitive to these unfavorable conditions, than is nodule growth. Maximum nitrogenase activity was observed during the podfilling stage; until 50–60 days after planting, nitrogenase activity was very low.     

Ecophysiological aspects of growth and nitrogen fixation inAzospirillum spp.

The nitrogenase activity ofAzospirillum spp. is efficiently regulated by environmental factors. InA. brasilense andA. lipoferum a rapid switch off of nitrogenase activity occurs after the addition of ammonium chloride. As in photosynthetic bacteria, a covalent modification of nitrogenase reductase (Fe-protein) is involved. InA. amazonense, a non-covalent mechanism causes only a partial inhibition of nitrogenase activity after ammonium chloride is added. In anaerobic conditions, nitrogenase reductase is also switched off by a covalent modification inA. brasilense andA. lipoferum. Short-time exposure ofAzospirillum to increased oxygen levels causes a partially reversible inhibition of nitrogenase activity, but no covalent modification is involved.Azospirillum spp. show variations in their oxygen tolerance. High levels of carotenoids confer a slightly improved oxygen tolerance. Certain amino acids (e. g. glutamate, aspartate, histidine and serine) affect growth and nitrogen fixation differently inAzospirillum spp. Amino acids may influence growth and nitrogen fixation ofAzospirillum in the association with plants.Azospirillum brasilense andA. halopraeferens are the more osmotolerant species. They utilize most amino acids poorly and accumulate glycine betaine, which also occurs in osmotically stressed grasses as a compatible solute to counteract osmotic stress. Nitrogen fixation is stimulated by glycine betaine and choline. Efficient iron acquisition is a prerequisite for competitive and aerotoleran growth and for high nitrogenase activity.Azospirillum halopraeferens andA. amazonense assimilate iron reasonably well, whereas growth of someA. brasilense andA. lipoferum strains is severely inhibited by iron limitation and by competition with foreign microbial iron chelators. However, growth of certain iron-limitedA. brasilense strains is stimulated by the phytosiderophore mugineic acid. Thus, various plant-derived substances may stimulate growth and nitrogen fixation ofAzospirillum.     

Host specificity of Rhizobium strains isolated from nitrogen-fixing trees and nitrogenase activities of strain GRH2 in symbiosis with Prosopis chilensis

Host plant specificity was examined in symbiosis between Rhizobium strains isolated from legume-tree root nodules and herbaceous or woody legumes from which they were isolated. Strain GRH2 isolated from Acacia cyanophylla formed effective nodules on Acacia, Prosopis and Medicago sativa as well. Nitrogenase activity, measured as acetylene reduction, of strain GRH2 in symbiosis with Prosopis chilensis was the highest (P 0.05) among the tropical legumes studied and was similar to those found for other associations involving herbaceous legumes. Relative efficiency of nitrogenase varied from 0.3 to near 1 during the light time of the photoperiod. However no hydrogen uptake activity was detected by the amperometric method used. Rhizobium strains GRH3, GRH5 and GRH9 isolated from A. melanoxylon, P. chilensis and Sophora microphylla, respectively, also showed a very low host-range specificity. All isolates were infective and effective on at least one of the herbaceous legumes tested. These data demonstrate the lack of specificity of Rhizobium strains isolated from nitrogen-fixing tree root nodules and that these strains can form effective nodules on herbaceous legumes.     

Seasonal variations of the sexual reproductive growth and nitrogenase activity (C2H2) in matureAlnus glutinosa

Summary The seasonal variations of the growth of sexual reproductive organs and of the nitrogenase activity (acetylene reduction) of root nodules are surveyed in mature field alders (Alnus glutinosa). The growth of female catkins—pollinated in February-early March—takes place chiefly from June to August and the growth of immature male catkins from July to September. The nitrogenase activity steadily shows two periods of high rate—the first from late April to early June, the second in September–October-and a summer period of low rate when the female catkins and the seeds achieve the most part of their growth.The seasonal fluctuations of thein vitro/in vivo nitrogenase activity ratio showing the supply of metabolic factors in the root nodules as a likely cause of the variations of thein vivo nitrogenase activity, the possible competition for photosynthate allocation between the production of sexual organs and the nitrogen-fixing capacity in mature field alders is discussed.     

Ammonia inhibition of nitrogenase activity in purple and green bacteria

Ammonia reversibly inhibits the nitrogenase activity not only in purple nonsulfur bacteria but in purple (Thiocapsa roseopersicina) and green (Chlorobium limicola forma thiosulfatophilum) sulfur bacteria as well.The complete inhibition of nitrogenase activity (acetylene reduction) is observed about 30 s after addition of NH ₄ ⁺ (2.5×10^-6 M) to cell suspensions. The pattern of ammonia inhibition of acetylene reduction in T. roseopersicina does not differ from the action of tetrabutylammonium and tetraphenylphosphonium (3 · 10^-6-5·10^-5 M) on nitrogenase activity of this bacterium.Simultaneously with the switch-off effect of NH ₄ ⁺ a considerable increase of ATP in cells of Rhodobacter sphaeroides and C. limicola f. thiosulphatophilum was observed.     

Nitrogenase activity in the non-heterocystous cyanobacterium Oscillatoria sp. grown under alternating light-dark cycles

The non-heterocystous cyanobacterium Oscillatoria sp. strain 23 fixes nitrogen under aerobic conditions. If nitrate-grown cultures were transferred to a medium free of combined nitrogen, nitrogenase was induced within about 1 day. The acetylene reduction showed a diurnal variation under conditions of continuous light. Maximum rates of acetylene reduction steadily increased during 8 successive days. When grown under alternating light-dark cycles, Oscillatoria sp. fixes nitrogen preferably in the dark period. For dark periods longer than 8 h, nitrogenase activity is only present during the dark period. For dark periods of 8 h and less, however, nitrogenase activity appears before the beginning of the dark period. This is most pronounced in cultures grown in a 20 h light – 4 h dark cycle. In that case, nitrogenase activity appears 3–4 h before the beginning of the dark period. According to the light-dark regime applied, nitrogenase activity was observed during 8–11 h. Oscillatoria sp. grown under 16 h light and 8 h dark cycle, also induced nitrogenase at the usual point of time, when suddenly transferred to conditions of continuous light. The activity appeared exactly at the point of time where the dark period used to begin. No nitrogenase activity was observed when chloramphenicol was added to the cultures 3 h before the onset of the dark period. This observation indicated that for each cycle, de novo nitrogenase synthesis is necessary.     

Drought-avoidant soybean germplasm maintains nitrogen-fixation capacity under water stress

Inoculated soybeans (Glycine max L. (Merrill)) were grown in controlled environments to evaluate the relationship between genotype and plant water status on nodule function, nitrogen assimilation, growth rates, and seed yield. Plants were grown under well-watered (WW) and water-stressed (WS) conditions during the linear pod-filling growth stage in sand culture using N-free nutrient solution. Dry matter and N accumulation were greater for the drought-adapted Plant Introduction 416937 (PI) than for Forrest, a commercially adapted genotype of similar phenology. These differences are attributed to: (i) more favorable internal water balance throughout the pod-filling period (higher total leaf water potential), (ii) higher photosynthetic function (more total leaf area and higher net carbon exchange rates), and (iii) stronger nodule function (larger nodule mass, greater specific and total nodule activity, and thus more nitrogen assimilation) for the PI than for Forrest. While Forrest out yielded the PI under WW conditions, the percentage reduction in seed mass per plant was less for the PI than for Forrest when both genotypes were exposed to desiccating conditions. The inference is that soybean germplasm with the capacity to maintain tissue turgidity, and thus leaf and nodule function, during reproductively-imposed desiccation may reduce the extent to which yield is compromised during drought. These findings have implications for the role of symbiotic nitrogen fixation in conserving yield under dry weather conditions.Abbreviations DAE Days After Emergence - NCE Net CO₂ Exchange - PI PI 416937 - SNA Specific Nodule Activity - TNA Total Nodule Activity - WS Water Stressed - WW Well Watered     

Evidence for an involvement of glutamine synthetase in regulation of nitrogenase activity in Rhodopseudomonas capsulata

In the presnet studies with whole cells and extracts of the photosynthetic bacterium Rhodopseudomonas capsulata the rapid inhibition of nitrogenase dependent activities (i.e. N₂-fixation acetylene reduction, or photoproduction of H₂) by ammonia was investigated. The results suggest, that the regulation of the nitrogenase activity by NH ₄ ⁺ in R. capsulata is mediated by glutamine synthetase (GS). (i) The glutamate analogue methionine sulfoximine (MSX) inhibited GS in situ and in vitro, and simultaneously prevented nitrogenase activity in vivo. (ii) When added to growing cultures ammonia caused rapid adenylylation of GS whereas MSX abolished the activity of both the adenylylated and unadenylylated form of the enzyme. (iii) Recommencement of H₂ production due to an exhaustion of ammonia coincided with the deadenylylation of GS. (iv) In extracts, the nitrogenase was found to be inactive only when NH ₄ ⁺ or MSX were added to intact cells. Subsequently the cells had to be treated with cetyltrimethylammonium bromide (CTAB). (v) In extracts the nitrogenase activity declined linearily with an increase of the ration of adenylylated vs. deadenylylated GS. A mechanism for inhibition of nitrogenase activity by ammonia and MSX is discussed.Abbreviations BSA bovin serum albumine - CTAB cetyltrimethylammonium bromide - GOGAT l-glutamine: 2-oxoglutarate amino transferase - GS glutamine synthetase - HEPES N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid - MSX l-methionine-d,l-sulfoximine     

Nitrogenase activity of two genotypes of Acacia mangium as affected by phosphorus nutrition

The specific nodulation, nitrogenase activity (acetylene reduction) and budgets of carbon allocation to respiration by nodulated roots were examined in two provenances of Acacia mangium Willd. grown in a glasshouse for 17 weeks to investigate the effects of soil phosphorus and genotypes of the host plant on symbiotic nitrogen fixation. Application of phosphorus (0–80 mg P kg^-1 soil) increased specific nodulation (g nodule dry weight g^-1 plant dry weight) of provenance Ma11 by two-fold and the percentage of nodulated root respiration allocated to nitrogenase by 50%, but had no effect on specific activity of nitrogenase or specific respiration coupled with nitrogenase activity. Improved phosphorus nutrition increased the specific nitrogenase activity of provenance Ma9 by 2-fold, the percentage of nodulated root respiration allocated to nitrogenase, and specific nitrogenase-linked respiration by 50%, respectively, but had no effect on the specific nodulation. The percentage of respiration coupled with nitrogenase activity in nodulated root respiration by provenance Ma9 was 60–70% higher than that in provenance Ma11, regardless of phosphorus levels applied. At the optimal level of phosphorus addition (10 mg P kg^-1 soil), provenance Ma9 had a lower dry mass than provenance Ma11. This was accompanied by a lower nodulated root respiration and a higher percentage of nodulated root respiration allocated to nitrogenase activity in provenance Ma9.     

Carbon discrimination,water-use efficiency,nitrogen and phosphorus nutrition of the host / mistletoe pair Eucalyptus behriana F. Muell and Amyema miquelii (Lehm. ex Miq.) Tiegh. at permanently low plant water status in the field

Summary Under conditions where both plants had permanently low water status, the mistletoe, Amyema miquelii (Lehm. ex Miq.) Tiegh., had lower nitrogen contents in leaf tissue than its host, Eucalyptus behriana F. Muell. The parasite transpired less than its host which is consistent with the hypothesis that mistletoe transpiration acts as a nitrogen gathering mechanism. Nitrogen and phosphorus contents were generally low in both plants; they were positively correlated, and mistletoes reduced nutrient contents of infested hosts. The carbon discrimination ratio, ¹³C (a measure of water-use efficiency) of each plant was within the range reported for other mistletoes and their hosts. Although it did not differ significantly between host and parasite it indicated lower water-use efficiency in the mistletoe. For the nitrogen content of host leaves the gradient within the pair, (¹³C), is much lower compared to the correlation given by Ehleringer et al. (1985). It is concluded that at permanently low water status on nitrogen and phosphorus deficient soils a water-saving strategy accompanied with slow growth is more appropriate for both mistletoe and host.