Biological Nitrogen Fixation in Pachyrhizus ahipa (Wedd.) Parodi

The patterns of plant growth and N₂ fixation capability in Pachyrhizusahipa (Wedd) Parodi inoculated with Bradyrhizobium ‘PachyrhizusSpec 1’ strains (Lipha Tech) were investigated in a zero-Nculture system under greenhouse conditions The P ahipa plantis day-neutral with respect to reproductive development Competitionoccurred between the two storage organs (legume and tuber) andprevented high tuber yield in P ahipa The symbiotic effectivenessof the association was high, as the profuse nodulation providedthe inoculated plants with adequate amounts of N Nodules werepresent throughout the cycle of P ahipa The change in rate ofN₂ fixation (RNF) and relative growth rate (RGR) was almostparallel during ontogenesis The developmental pattern of N₂fixation activity revealed that 65% of total N₂ fixation occurredafter N began to accumulate in the reproductive (pod wall plusseed) tissue During pod filling allocation of N compounds tothe seeds exceeded N₂ fixation, the pod walls being the primarysource of redistributed N, followed by the leaves. Pachyrhizus ahipa (Wedd) Parodi, ahipa, tuber crop, dinitrogen fixation, dry matter, N partitioning, reproductive growth     

Nitrogen Fixation in Wild Legumes

Isolations of the rhizobia from nodules of field plants of Ulexeuropaeus and Medicago lupulina have been examined for effectivenessin fixation of nitrogen by inoculating them into host plantsgrowing in a medium free of combined nitrogen. The results indicatethat vigorous fixation may be presumed to occur in associationwith field plants of the Ulex species, but that fixation inMedicago is probably decreased by the presence of ineffectiverhizobia in some nodules. The general importance of fixationin wild legumes is discussed, and it is pointed out that dueattention must be paid to non-legume genera with root nodules.These genera are locally abundant, and in former periods werestill more prominent in some regions of the world.     

Non-Autotrophic CO2 Fixation and Drought Tolerance in Mosses

Cratoneuron filicinum, a drought-sensitive moss, and Tortularuralis, a drought-tolerant moss, fix CO₂ non-autotrophicallyat a rate of about 1.2 and 2.2 µmol h^–1 g^–1dry wt. respectively. During drying, T. ruralis fixes CO₂ atan undiminished rate until the tissue loses about 60% of theinitial fresh weight. Thereafter, CO₂ fixation rapidly declinesto zero. Dark CO₂ fixation by C.filicinum declines steadilyduring the dehydration period. On rehydration, dark CO₂ fixationis resumed immediately in T. ruralis but not in C.filicinum.When dried T. ruralis is equilibrated with an atmosphere ofnearly 100% relative humidity, its weight increases to about40% of the original fresh weight and dark CO₂ fixation resumesat a rate about 60% of the fresh moss. In C.filicinum thereis only a small increase in weight and little CO₂ fixation inthe dark. The non-autotrophically fixed carbon, in both mossesstudied, is incorporated into amino acids (more than 60% ofthe total, mainly into aspartate, alanine and glutamate) andorganic acids (less than 40% of the total, mainly into malate).It is suggested that on rehydration immediate availability ofNADPH, known to be produced by transhydrogenation from NADHduring dark CO₂ fixation, may be an important factor in therepair of drought-induced cellular damage by reductive biosynthesisof membrane components and other cellular constituents. Key words: Mosses, Dehydration, Rehydration, Dark CO₂ fixation, Amino acids, Organic acids, NADPH, Drought tolerance.     

Young Reproductive Structures Promote Nitrogen Fixation in Soya Bean

In many legumes the transition from the vegetative to the reproductivephase of development is associated with a marked increase inthe rate of symbiotic nitrogen fixation. In soya bean [Glycinemax (L.) Merr.), the removal of reproductive parts at differentstages of their development showed that the increase in nitrogenfixation rate was primarily due to the presence of flower buds.The increase in the fixation rate of intact reproductive plantswas accompanied by a rapid increase in the weight of noduleson lateral roots and it is suggested that these nodules areresponsible for much of the nitrogen fixation which occurs duringreproductive growth. Maintaining plants in the vegetative stateprovided evidence which suggests that it is the flower budsand not the flowering stimulus which are responsible for theincrease in fixation rate. The marked effects on vegetativegrowth of removing reproductive parts suggests that the mechanisminvolved in the promotion of nitrogen fixation may be hormonal. Glycine max (L.) Merr., soya bean, nitrogen fixation     

Evaluation of nitrogen fixation in the diatom genus Rhizosolenia Ehr. in the absence of its cyanobacterial symbiont Richelia intracellularis Schmidt

Aggregates of two species of Rhizosolenia (R. castracanei andR. imbricata var. shrubsolei) have been reported to containendosymbiotic bacteria and to fix nitrogen. The general importanceof this process to the genus is not known, although of greatpotential significance. Clonal cultures of five species of Rhizosolenia(R. alata, R. bergonii, R. calcar-avis, R. imbricata var. shrubsoleiand R. setigera) were examined for nitrogen fixation using acetylenereduction and compared to cyanobacterial controls. Mecososmaggregates of R. imbricata var. shrubsolei were tested, as wellas chains of R. debyana hand-collected using SCUBA. The Rhizosoleniaspecies did not contain the endosymbiotic cyanobacterium Richeliaintracellularis and were not examined for the presence of endosymbioticbacteria. Nitrogen fixation was not found in the Rhizosoleniaspp.; in contrast, the diazotrophic cyanobacterial controlsOscillatoria erythraea and Anabaena sp. reduced acetylene atsignificant rates. The absence of nitrogen fixation in the Rhizosoleniaspp. suggests diazotrophy by endosymbiotic bacteria is not widespreadin the genus Rhizosolenia, nor is it necessary for survivalof these large diatom cells in nutrient-poor, near-shore waters.     

Interrelationship Between Plant Developmental Stage, Plant Growth Rate, Nitrate Utilization and Nitrogen Fixation in Hydroponically Grown Soybean

The growth rate of the indeterminate soybean plant [Glycinemax (L.)Merr.] slows as it proceeds from vegetative phase intoreproductive growth. Yet, the well-nodulated plant acquiresmost of its nitrogen during reproductive growth. Thus, the interrelationshipbetween plant developmental stage and nitrogen fixation wasexamined. It is shown that, regardless of the age of the hydroponicallygrown soybean plant at the time of its inoculation with Bradyrhizobiumjaponicum, the highest rate of nitrogen fixation occurs duringthe pod-filling stage (R5). Nevertheless, maximum total nitrogenfixation is generally achieved when inoculation occurs at thefull-bloom stage (R2). It is shown, however, that flower budsand flowering are not responsible for the enhanced nodulationand nitrogen fixation. Rather, the data suggest that the onsetof rapid nodulation occurs soon after the initiation of thedevelopmentally programmed drop in foliar nitrate reductaseactivity. The ensuing increase in nitrogen fixation providesthe plant with much of its needed nitrogen and hence stimulatesplant mass accumulation during pod-fill. It is suggested thatnitrogen fixation enhances growth of the soybean plant by increasingits net photosynthetic efficiency during reproductive growthand by providing the needed nitrogen at the appropriate timefor maximum seed growth. Key words: Glycine max, nitrate, nitrogen fixation, nodulation     

Growth and nitrogen assimilation in nodules in response to nitrate levels in Vicia faba under salt stress

This study analyses the effects of salt on the effective symbiosisof faba bean (Vicia faba L. var. minor cv. Alborea) and salt-tolerantRhizobium leguminosarum biovar. viciae strain GRA19 grown withtwo KNO₃ levels (2 and 8 mM). The addition of 8 mM KNO₃ to thegrowth medium increases plant tolerance to salinity even witha concentration of 100 mM NaCl. This KNO₃ level in control plantsreduced the N₂ fixation. For 2 and 8 mM KNO₃ the plants treatedwith NaCl reduced N₂ fixation to identical values. The activityof the enzymes mediating ammonium assimilation in nodules (GS,NADH-GOGAT and NADH-GDH) was decreased by high KNO₃ levels.The results show that NADH-GOGAT activity was more markedlyinhibited than was GS activity by salinity, therefore NADH-GOGATlimits the ammonium assimilation by nodules in V. faba undersalt stress. The total proline content in the nodule was notrelated to salt tolerance and thus does not serve as a salttoleranceindex for V. faba. Key words: Glutamate synthase, glutamine synthetase, N₂ fixation, nitrate, salinity     

Inhibition of Nitrogen Fixation in Non-Legume Root Nodules by Hydrogen and Carbon Monoxide

The effect of the presence of hydrogen and of carbon monoxideon the fixation of nitrogen in detached root nodules of non-legumeshas been studied, fixation being measured by the use of ¹⁵N.Parallel tests on legumes (pea and soya bean) have been included.Fixation in the nodules of Casuarina, Alnus, and Myrica is inhibitedin the presence of substantial proportions of hydrogen, to adegree resembling that shown in legumes. Fixation in Alnus andMyrica is arrested in the presence of small proportions of carbonmonoxide, and here again the sensitiveness is of the same orderas in legumes.     

The Fixation of Nitrogen associated with the Root Nodules of Myrica gale L., with Special Reference to its pH Relation and Ecological Significance

The paper deals with the fixation of nitrogen by nodulated plantsof Myrica gale under experimental conditions, the fixation being,in the first year of development, of a magnitude comparableto that of legumes under similar conditions. The effect of acidityon the fixation has been investigated. The Myrica organism ismarkedly adapted to acid conditions. The observations are correlatedwith field data in Britain and help to explain the distributionand habits of the species.     

Validity of N2 fixation rate measurements in marine Oscillatoria (Trichodesmium)

No measurable differences in Trichodesmium nitrogenase activitywere observed between colonies collected by diving and incubatedunder ultra-clean conditions compared with those collected andincubated using standard techniques. Measurements were madein the northeastern Caribbean Sea, near the Bahama Islands andin the Sargasso Sea. Surprisingly, mean rates of ethylene productionwere high relative to most previous in situ measurements onTrichodesmium. The calculated cellular N doubling times (viaN₂ fixation) ranged from 1.13 days in the northeastern CaribbeanSea, 1.48 days in the Sargasso Sea to 1.8 days near the BahamaIslands. A comparison of these doubling times with those inthe literature illustrates the high variability in rate of N₂fixation by Trichodesmium. From this study, we conclude thatthe often observed slow rates of N₂ fixation are valid. Populationsof Trichodesmium can probably remain within the water columnat low growth rates via gas vesicles, which keep the colonysuspended, and low grazing rates by herbivores.     

The Rate of Photosynthesis in Isolated Chloroplasts

Chloroplasts were isolated using aqueous and nonaqueous procedures.Aqueous chloroplasts lost approximately 50 per cent, of theirsoluble proteins during isolation. Nonaqueous chloroplasts retainedall their soluble enzymes, but lost their ability to performthe light reactions of photosynthesis. It was possible to reconstitutea chloroplast system of higher activity by adding soluble enzymesfrom nonaqueous chloroplasts to protein-deficient aqueous chloroplasts.The properties of the reconstituted chloroplast system wereas follows: 1. The CO₂ fixation rate of the reconstituted chloroplast system( 4 µM./. chlorophyll/hr.) was 3–4 times that ofthe aqueous chloroplasts ( I µM./. chlorophyll/hr.). Thefixation of aqueous chloroplasts isapparently limited in partby lack of soluble enzymes. 2. During light-fixation, the reconstituted chloroplast systemaccumulated PGA. This indicates that the reduction of PGA totriosephosphate is a rate-limiting step in this system. 3. It was possible to increase the CO₂ fixation to 12 µM.CO₂/mg. chlorophyll/ hr. by addition of ATP and TPNH to thesystem, but the reduction of PGA was still rate-limiting. 4. Further increase in the fixation rate was obtained by concentratingthe reaction mixture. Part of the striking differences of theCO₂-fixing capabilities of chloroplasts in vivo and in vitrois caused by dilution effects. Extrapolation of the dilutioneffect to the protein concentration which exists in chloroplastsyields a CO₂ fixation rate of approximately 30 µM./mg.chlorophyll/hr. 5. Inhibitors which are located in vivo outside the chloroplastsaffect the CO₂ fixation in vitro. 6. Under consideration of the examined factors which influencethe CO₂ fixation of isolated chloroplasts, it is possible toraise the fixation from approximately 1 per cent, to at least15 per cent, of the fixation in vivo.     

Pathway of dark inorganic carbon fixation in two species of diatoms: influence of light regime and regulator factors on diel variations

Planktonic algae submitted to vertical mixing with a short periodicitycommute many times a day from low to high irradiance levels.To study the influence of this light periodicity, two diatoms,Skeletonema coslatum and Nitzschia turgiduloides, were cultivatedunder alternating conditions of 2 h light/2 h dark (2 h/2 h),simulating vertical mixing in the natural environment. Two otherlight regimes were used: continuous light (CL) and alternatecycles of 12 h light/12 h dark (12 h/12 h). Products synthesizedin the dark by S.costmum during 60 s incubation for 2 h/2 hculture or during 5 min for 12 h/12 h culture were determined.They were essentially sugars, malate, aspartate and glyceratefor 2 h/2 h cells and 12 h/12 h cells taken at the beginningof the light period. In contrast, 12 h/12 h cells taken duringthe darkness or in the middle of the light period and set inthe dark synthesized only amino acids. Our results corroborateprevious reports on dark CO₂ fixation via phosphoenolpyruvatecarboxykinase (PEPCKase, enzyme allowing the fixation of CO₂on PEP and the synthesis of amino acids) with involvement ofa substrate synthesized during the light period, but demonstratethat incorporation also occurs by the C-3 pathway (pathway responsiblefor the major CO₂ fixation in the light) in the very early stagesof the dark period. Another important result highlighted bythis study is the appreciable rate of dark fixation: on average6.7, 8.3 and 12.7% of photosynthesis at saturating photon fluxdensity for N.turgiduloides cultivated under 2 h/2 h, CL and12 h/12 h regime respectively and nearly 12% for S.costatumin the 2 h/2 h light regime. Variation of dark fixation wasinvestigated as a function of hour in the two species. Skeletonemacostatum cells submitted to the 2 h/2 h cycle show a constantrate of light-independent assimilation throughout the day. Bycontrast, both N.turgiduloides grown under the 12 h/12 h or2 h/2 h regime and S.costatum cultured under the 12 h/12 h cycleundergo fluctuations in the rate of dark fixation over the light/darkcycle. The mean dark fixation rate is controlled by the lengthof the photoperiod or the frequency of light fluctuations, dependingon species. We argue that this phenomenon must be taken intoconsideration in primary production calculations. Dependingon whether they are synthesized at the beginning or at the endof the light period, products are somewhat different and therate of fixation varies. This leads us to suggest that the pathwayof dark fixation may be regulated by at least two factors: amountof available substrate and enzyme (RuBPCase and PEPCKase) activityand/or amount.     

Photosynthetic Changes in the Inducible CAM Plant Sedum telephium L. Following the Imposition of Water Stress. 1. General Characteristics

The effects of water stress (drought) on the pattern of photosynthesisin Sedum telephium have been determined. Well-watered plantsexhibit a weak-CAM pattern, with substantial CO₂ fixation inthe day, a low level of CO₂ fixation at night, high daytimestomatal conductance with a lower conductance at night, andno diurnal fluctuation in acid content. Imposition of water-stress causes a switch from weak-CAM toa full-CAM mode of photosynthesis, as indicated by cessationof daytime CO₂ fixation, a marked increase in night-time CO₂fixation, very low daytime stomatal conductance, increased night-timeconductance and significant diurnal fluctuations in acid content. Sedum telephium, CAM, CO₂ fixation, drought, malate, photosynthesis, water stress     

Effects of Aldehyde Fixation on Adenosine Triphosphatase and Peroxidase Activities in Maize Root Tips

The effects of aldehyde fixation of excised roots and subcellularfractions of Zea mays have been studied in relation to the preservationof ATP-ase and peroxidase activities. Total peroxidase was littleaffected by either formaldehyde or glutaraldehyde whereas ATP-aseshowed considerable loss of activity, particularly with glutaraldehyde.The activity remaining after fixation was dependent on boththe concentration of fixative and the pH of fixation. Subcellularfractions differed in their response to fixation with the cellwall fraction generally showing higher retention of activitythan the mitochondrial or microsomal fractions.     

Nitrogen Fixation in the Canopy of Temperate Forest Trees: A Re-examination

¹⁵N₂ studies and acetylene reduction assays of leaves and shootsof Douglas fir and other forest trees do not confirm previousreports that extensive nitrogen fixation occurs on leaf surfacesand it is concluded that the importance of nitrogen fixationin the canopy of forest trees has been exaggerated. The presenceof nitrogen-fixing bacteria on the leaves of trees is confirmed,however, and they have been identified as Enterobacter agglomerans,Clostridium butyricum and Bacillus sp. Their distribution onleaves is fortuitous since dead oak leaves and artificial leavesbecome colonized to the same extent as living oak leaves. nitrogen fixation, acetylene reduction, Enterobacter agglomerans, Clostridium butyricum, Bacillus sp, Douglas fir, Pseudotsuga menziensii, larch, Larix x oak, Quercus petraea.     

Seasonality of Rhizome and Shoot Production and Nitrogen Fixation in Lotus uliginosus under Upland Conditions in South-west Scotland

Development, growth and nitrogen fixation were monitored over3 years in marsh birdsfoot trefoil (Lotus uliginosus Schkuhr),at an upland site in south-west Scotland Plants exhibited markedseasonality of growth, with production of both aerial shootsand rhizome reaching a peak in September and October Renewedaerial shoot growth in spring was initiated mainly from nodeson rhizomes formed in the previous autumn Nitrogen fixationwas measured in soil cores using the acetylene reduction techniqueFixation commenced in early June, coinciding with increasingsoil temperature, and thereafter rose to a peak in mid-summerThe subsequent decline in fixation was associated with increasingrainfall, decreasing soil temperature, flowering and the onsetof rhizome production Lotus uliginosus Schkuhr, marsh birdsfoot trefoil, seasonality, rhizome production, nitrogen fixation, acetylene reduction     

An Examination of some Symbiotic Systems for Fixation of Nitrogen

By the use of ¹⁵N the occurrence of fixation of elemental nitrogenhas been demonstrated in the lichens Collema granosum and Leptogiumlichenoides (both of which contain Nostoc as a symbiont), andalso in the liverwort Blasia pusilla (which has Nostoc-containingcavities in the thallus). The circumstances indicate that thefixation in these instances should be attributed to the Nostoc. No evidence of fixation was obtained in similar isotopic testson the mycorrhizal roots of intact plants of Calluna vulgarisand of Pinus sylvestris, although these, more especially inthe case of Calluna, have been held by some previous authorsto be nitrogen-fixing.     

Responses of N2 Fixation-linked Respiration to Host-plant Energy Status in White Clover Acclimated to a Controlled Environment

Ryle, G. J. A., Powell, C. E. and Gordon, A. J. 1988. Responsesof N₂ fixation-linked respiration to host-plant energy statusin white clover acclimated to a controlled environment.—J.exp. Bot. 39: 879–887. Single plants of white clover, acclimated to a controlled environmentand dependent for nitrogen on N₂ fixation in their root nodules,were darkened, defoliated or exposed to enhanced CO₂ levelsto establish the quantitative relationships between the photosynthesisof the host plant and the N₂ fixation metabolism of root nodules. The nodule respiration associated with N₂ fixation (FLR) declinedrapidly to 10–15% of its normal rate following plant darkeningearly in the photoperiod. Darkening at progressively later intervalsduring the photoperiod demonstrated a positive, apparently linearrelationship between duration of illumination and total FLRduring the photoperiod and the following night period. Completeor partial defoliation reduced FLR according to the leaf arearemoved: again, there was a strong positive correlation betweencurrent rate of photosynthesis, whether of defoliated or undefoliatedplants and the FLR of root nodules. Doubling the current rateof photosynthesis, by enhancing CO₂ levels around the shoots,promoted FLR within 1–2 h when plants were stressed bylack of light. However, enhanced CO₂ levels increased FLR onlyslowly over a period of several hours in plants entrained tothe normal growing conditions. It is concluded that, in these plants acclimated to a uniformand favourable controlled environment, the supply and utilizationof photosynthetic assimilate in N₂ fixation was finely balancedand quantitatively linked during a single diurnal period andthat nodule functioning was not depressed by lack of energysubstrate. Key words: White clover, N₂ fixation, photosynthesis.     

Metabolic changes in soybean nodules after inhibition of nitrogen fixation by treatment with oxygen

Metabolites that accumulated in soybean [Glycine max (L.) Merr.]nodules after inhibition of nitrogen fixation were analysedto determine what carbon compounds the bacteroids might obtainfrom their host. Exposure of roots of intact soybean plantsto 100% O₂ for 5 min caused a decrease in acetylene reductionactivity within 10 min and then the activity recovered onlyslowly. Analysis of carbohydrates, organic acids, volatile compoundsand amino acids in extracts of nodules revealed that succinate,malate and alanine all accumulated within 10 min after treatmentwith O₂. The concentrations of sucrose, acetone, tyrosine, valine,isoleucine, leucine, and ornithine in the nodules increasedslowly after such treatment. The results are discussed in termsof carbon sources for supporting nitrogen fixation of soybeanbacteroids. Key words: Glycine max, carbon metabolism, nitrogen fixation, nodules     

Fixation of Elemental Nitrogen by Marine Blue-green Algae

Three blue-green algae, Calothrix scopulorum, Nostoc entophytum,and Oscillatoria brevis, isolated from the upper littoral andsupralittoral fringe of the sea-shore were obtained in pureculture and tested for fixation of elemental nitrogen. Appreciablefixation by Calothrix and Nostoc was detected, a proportionof the total nitrogen fixed being liberated into the culturemedium. There was no evidence of fixation by Oscillatoria. Thisappears to be the first evidence that blue-green algae isolatedin pure culture from marine habitats fix nitrogen.