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.     

蓝藻Anabaena 7120排氨的研究

蓝藻Anabaena7120在光下和暗中的排氨量显著不同,暗中比光下高。光下加光合抑制剂和光照强度减弱均促进蓝藻排氨。抑制固氮的CO和O_2也削弱蓝藻排氨。无论是在空气、分子氮和氩气中,还是在光和暗条件下,加MSX都促进蓝藻排氨,MSX和DCMU—同加入时,蓝藻排氨量更高。     

Nitrogen Fixation by the Blue-green Alga Anabaena flos-aquae A-37

Nitrogen fixation has been demonstrated in a bacteria-free culture of a semithermophilic blue-green alga, Anabaena flos aquae A-37, employing the criterion of an increase in cellular nitrogen as a basis for determining nitrogen fixation. This is apparently the first time that nitrogen fixation has been demonstrated in a pure culture of a blue-green alga which grows at 40 C and which produces abundant quantities of an extracellular heteropolysaccharide.     

The Respiratory Costs of Nitrogen Fixation in Soyabean, Cowpea, and White Clover: II. COMPARISONS OF THE COST OF NITROGEN FIXATION AND THE UTILIZATION OF COMBINED NITROGEN

Plants of soyabean, cowpea, and white clover were grown singlyin pots in Saxcil growth cabinets at 23/18 °C, 30/24 °C,and 20/15 °C, respectively, until seed maturation or for85 d (white clover). Two populations were produced within eachspecies: one population effectively nodulated and wholly dependentfor nitrogen on fixation in the root nodules, and a second populationcompletely lacking nodules but receiving abundant nitrate nitrogen.In each species, the two populations were compared in termsof rate of gross photosynthesis, rate of shoot respiration,and rate of root respiration. Source of nitrogen had littleor no effect on rate of photosynthesis or shoot respiration.In contrast, the rate of respiration of the nodulated rootsof plants fixing their own nitrogen was greater, sometimes two-foldgreater, than that of equivalent plants lacking nodules andutilizing nitrate nitrogen. This superiority in terms of rateof root respiration was generally confined to the period ofintense nitrogen fixation. An analysis of the magnitude of thisrespiratory burden in terms of daily photosynthesis indicatesthat, in all three legumes, plants fixing their own nitrogenrespire 11–13% more of their fixed carbon each day thanequivalent plants lacking nodules and utilizing nitrate nitrogen.     

Effect of Interactions Among Algae on Nitrogen Fixation by Blue-Green Algae (Cyanobacteria) in Flooded Soils

Nitrogen fixation (C₂H₂ reduction) by algae in flooded soil was limited by interactions within the algal community. Nitrogen fixation by either indigenous algae or Tolypothrix tenuis was reduced severalfold by a dense suspension of the green alga Nephrocytium sp. Similarly, interactions between the nitrogen-fixing alga (cyanobacterium) Aulosira 68 and natural densities of indigenous algae limited nitrogen-fixing activity in one of two soils examined. This was demonstrated by developing a variant of Aulosira 68 that was resistant to the herbicide simetryne at concentrations that prevented development of indigenous algae. More nitrogen was fixed by the resistant variant in flooded soil containing herbicide than was fixed in herbicide-free soil by either the indigenous algae or indigenous algae plus the parent strain of Aulosira. Interference from indigenous algae may hamper the development of nitrogen-fixing algae introduced into rice fields in attempts to increase biological nitrogen fixation.     

The Respiratory Costs of Nitrogen Fixation in Soyabean, Cowpea, and White Clover: I. NITROGEN FIXATION AND THE RESPIRATION OF THE NODULATED ROOT

Soyabean, cowpea, and white clover, inoculated with effectiverhizobia, were grown singly with a standard mineral nutritionand light regime in controlled environments until seed maturation(in soyabean and cowpea) or late vegetative growth (white clover).Day/night temperature regimes were 23/18, 30/24, and 20/15 °Cin soyabean, cowpea, and white clover, respectively. The respiratorylosses of CO₂ from the nodulated root systems were studied inrelation to the concurrent rate of fixation of atmospheric nitrogen.Despite differences in development, the effectiveness of thesymbioses, and the temperature of growth, all three legumesexhibited similar respiratory losses from nodulated roots perunit of nitrogen fixed. During intense nitrogen fixation, theaverage respiratory losses for the three legumes varied between6·3 and 6·8 mg C mg^–1 N; within each species,the losses varied more widely at different stages of development.These respiratory burdens reflect the total cost to the plantof the nodule/nitrogen fixation syndrome including the subtendingroots. The results are discussed in relation to the respiratoryeffluxes from nodules and roots, and to biochemical investigationsof the costs of nitrogen fixation.     

Growth and Nitrogen Fixation of Phaseolus vulgaris L. at Two Irradiances II. Nitrogen Fixation

Phaseolus vulgaris cv. Glamis plants grown at 7 and 28 W m^–228 W m^–2 in controlled environment cabinets showed copiousnodulation and high levels of acetylene reducing activity. Earlydifferences in nodulation were apparent before differences inphotosynthesis and were attributed to an effect of far-red lighton nodule development. Total plant nitrogen content was greater at 28 W m^–2 thanat 7 W ^–2 but nitrogen content as a percentage of d. wtwas greater at the lower irradiance level. Total acetylene reducing activity (nmol. min^–1 root^–1)was greater at 28 W m^–2 than at 7 W ^–2, but therewas no irradiance effect on specific activity (nmol. min^–1g d. wt of pink nodules^–1 or nmol. min^–1 pink nodule^–1). Transfer of 40-day-old plants from 7 W m^–2 to 28 W m^–2resulted in increased nodulesize(due toincreased size of infectedcells), accompanied by increased total, but not specific, acetylenereducing activity. Transfer of plants from 28 W m^–2 to 7 W m^–2 resultedin a fall of total acetylene reducing activity within 24 h,and senescence of large nodules. Specific acetylene reducingactivity was unaffected The results are interpreted as an effect of light on the productionof nitrogen fixing tissue, rather than on nitrogenase activity.     

Nitrogen Fixation by Thermophilic Blue-Green Algae (Cyanobacteria): Temperature Characteristics and Potential Use in Biophotolysis

Thermophilic, nitrogen-fixing, blue-green algae (cyanobacteria) were investigated for use in biophotolysis. Three strains of Mastigocladus laminosus were tested and were found to be equally effective in biophotolysis as judged by nitrogenase activity. The alga, M. laminosus NZ-86-m, which was chosen for further study, grew well in the temperature range from 35 to 50°C, with optimum growth at 45°C, at which temperature acetylene reduction activity was also greatest. The maximum tolerable temperature was 55°C. Acetylene reduction activity was saturated at a light intensity of 1 × 10⁴ ergs cm⁻² s⁻¹. Atmospheric oxygen tension was found to be slightly inhibitory to acetylene reduction of both slowly growing and exponentially growing cultures. Nonsterile continuous cultures, which were conducted to test problems of culture maintenance, could be operated for 2 months without any significant decrease in nitrogenase activity or contamination by other algae. Nitrogen-starved cultures of M. laminosus NZ-86-m produced hydrogen at comparable rates to Anabaena cylindrica. The conversion efficiency of light to hydrogen energy at maximum rates of hydrogen production was 2.7%.     

Growth of Two Soybean Shoots on a Single Root: EFFECT ON NITROGEN AND DRY MATTER ACCUMULATION BY SHOOTS AND ON THE RATE OF NITROGEN FIXATION BY NODULATED ROOTS

Soybean stems were grafted between the first and second nodes6 weeks after planting. Three or 5 weeks after grafting, oneroot system was cut from grafted plants, doubling the shoot:root ratio. This technique was applied to plants grown in sandculture and supplied with an excess of water and minerals butwith no combined nitrogen so that doubling the shoot:root ratiogreatly increased the requirement for fixed nitrogen. When theshoot:root ratio was doubled during the flowering or seed formationstages, there was no statistically significant effect on totalnitrogen or dry matter of shoots compared to non-grafted controlsat maturity. The period between 2 and 21 d after doubling theshoot:root ratio was studied to determine changes in root weight,nodule weight, and rate of nitrogen fixation (acetylene reduction).Weight of roots and nodules increased relative to controls (graftedplants with one shoot per root) after about 1 week. The rateof acetylene reduction per gram of nodule was 75 per cent greaterwith roots having two shoots than with control roots 2 d afterdoubling the shoot:root ratio. Acetylene reduction per noduledeclined to near control rates as nodule weight increased, butroots with two shoots maintained a 60 to 70 per cent greaterrate of acetylene reduction per root for 2 weeks. The resultsindicated that nodulated soybean roots are capable of fixingnitrogen at rates greater than those which normally prevail.     

The Effects of Moisture on Acetylene Reduction by Mats of Blue-green Algae in Sub-tropical Grassland

During the dry winter months desiccation is the main factorpreventing nitrogen fixation by mats of blue-green algae inkikuyu lawns. In the wetter summer months a few days of sunshinewithout rain will dry out the algal mats and depress nitrogenfixation unless the algae are protected by swards of grass.Dry mats resume nitrogen fixation within an hour of becomingdamp and attain their normal rate of fixation within 5 h. Maximumfixation rates occur at 100 per cent relative humidity and withsoil moisture contents of 22 to 42 per cent. Dry algal matsare unable to take up sufficient water as vapour (even in saturatingconditions) to permit resumption of acetylene reduction. Algal-dominatedsoils show greater water retention than adjacent bare soils.     

Nitrogen Fixation by Subterranean Clover at Varying Stages of Nodule Dehydration: II. EFFICIENCY OF NITROGENASE FUNCTIONING

Changes in nitrogenase activity (C₂H₂ reduction and H₂ production),nodulated root respiration and the efficiency of nitrogenasefunctioning were measured in response to progressive dehydrationof nodules on intact well-watered plants of subterranean clover(Trifolium subterraneum L.) cv. Seaton Park. The nodulated rootsof vegetative plants grown to the 14-leaf stage were incubatedin a gas exchange system through which a continuous dry airstreamwas passed over an 8 d period. The root tips were immersed inan N-free nutrient solution during this time so that water andion uptake was unimpeded. The decline in nodulated root respirationresulting from nodule drying was associated with a continualreduction in respiration coupled to nitrogenase activity. Asnodule water potential (_nod) decreased, the proportion of totalnodulated root respiration which was nitrogenase-linked declinedfrom 50% (day 1) to 33% (day 8). This was accompanied by a 79%reduction in specific nitrogenase activity (from 3.79 to 0.81umol C₂H₄ g^–1 nodule dry weight min^–1). Nodule dehydrationalso induced a decline in hydrogen (H₂) production in air. Therelative decline in hydrogen production exceeded that of acetylenereduction activity and this resulted in an increase in the relativeefficiency of nitrogenase functioning. However, the carbon costof nitrogenase activity progressively increased above 2.0 molCO₂ respired per mol C₂H₄ reduced as rood decreased below –0.4to –0.5 MPa. Consecutive measurements of the rates ofhydrogen evolution, ¹⁵N₂ fixation and acetylene reduction activityon intact unstressed plants resulted in a C₂H₄/N₂ conversionfactor of 4.08 and an electron balance of 1.08. These resultsindicated that the pre-decline rate of acetylene reduction activitymeasured in a flow-through system provided a valid measure ofthe total electron flux through nitrogenase. Key words: Subterranean clover, dehydration, efficiency, nitrogenase activity