Effect of low rhizosphere oxygen on growth, nitrogen fixation and nodule morphology in lucerne

Oxygen shortage in soils can occur following a wide range of natural circumstances, affecting the plant's physiology. In this paper the performance of nodulated lucerne plants under severe hypoxia is examined and the mechanisms involved to achieve this adaptation are discussed. Nodulated lucerne plants ( Medicago sativa L.) were grown with their rooting medium exposed to 1 or 21 kPa oxygen. Final yield, as expressed on a shoot dry weight basis, was unaffected but root and nodule dry weights were reduced by 50%. Water content in roots and nodules was higher at 1 kPa as a result of the formation of aerenchyma. Specific acetylene reduction activity was higher in hypoxic nodules as a consequence of modified nodule structure, although they were more sensitive to the presence of acetylene or nitrate. Root respiration was insensitive to changes in external oxygen supply, therefore providing adequate support for mineral uptake. Nodule respiration rates were 5 times higher in control plants when measured as CO₂ evolution, whereas no differences were observed in O₂ uptake. It is suggested that adaptation of nodulated lucerne to low oxygen concentrations involves changes in photosynthate allocation and nodule morphology, which provide a more efficient nitrogen fixation.     

ALLELOPATHIC GROWTH INHIBITION OF NODULATED AND UNNODULATED ALNUS CRISPA SEEDLINGS BY POPULUS BALSAMIFERA

Water extracts of fresh leaves, buds, and leaf litter of Populus balsamifera (balsam poplar) were tested at different dilutions for allelopathic effects on nodulation, nitrogenase activity and growth of nodulated green alder (Alnus crispa) seedlings, and on growth of unnodulated green alder seedlings. All extracts inhibited height growth, root elongation and dry weight increment of nodulated and unnodulated green alder seedlings to some degree during a 2-month experiment. Foliar nitrogen content of both nodulated and unnodulated seedlings was significantly lower in extract treated plants than in controls. Growth inhibition was about 25% less in nodulated than in unnodulated seedlings. The number of nodules per plant in seedlings treated with any balsam poplar extract was only 51% that of control plants. Acetylene reduction by seedlings treated with bud and leaf litter extracts indicated a decrease of 62% compared to controls. Growth inhibition was not mediated by pH or osmotic effects of the extracts. The possible ecological significance is discussed.     

How does plant population density affect the biomass of Ravenna grass?

Ravenna grass, Tripidium ravennae (L.) H. Scholz, is known to produce an abundance of biomass, but how plant density affects its biomass potential remains unknown. The objectives were to determine the effects of plant density on biomass yield; plant growth traits; biomass?carbon, nitrogen, and ash concentrations; heating value; nitrogen removal; and sucrose concentration in leaves and culms. The treatments consisted of five plant densities (1,250; 2,500; 5,000; 10,000; and 20,000 plants per hectare) in a randomized complete block design with four blocks. Plots were nonirrigated, unfertilized, and harvested once during the dormant season each year. Data were collected from 2015?2019. Dependent variables that varied with plant population density (p < .05) were biomass yield, number of reproductive culms per plant, reproductive culm diameter, reproductive culm sucrose concentration, and nitrogen removal with biomass. Biomass yield ranged from 5.6 to 16.3 Mg/ha for plant densities of 1,250–20,000 plants per hectare, respectively. Combined over years, nonlinear regression of the data showed the equation for biomass yield to plateau at 16.2 Mg/ha at a plant density of 10,640 plants per hectare. As plant density increased, the number of reproductive culms per plant, culm diameter, and culm sucrose concentration significantly decreased. At 1,250 plants per hectare, the number of reproductive culms per plant, culm diameter, and culm sucrose averaged 70, 10.2 mm, and 63.2 g/kg, respectively. Nitrogen removed with biomass significantly increased as biomass yield increased with plant density. At a density of 10,000 and 20,000 plants per hectare, the amount of nitrogen removed annually in the harvested biomass averaged 88 kg/ha. The data suggest that 10,000 plants per hectare would produce the greatest annual biomass yields; however, research is needed to determine the nutrient requirement for Ravenna grass to sustain biomass production at that density.     

A functional relationship between leghaemoglobin and nitrogenase based on novel measurements of the two proteins in legume root nodules

A combination of physiological and structural measurements made on nodulated cowpea and soybean plants cultured with roots in different pO(2) permitted the expression of data in various ways. Values of leghemoglobin concentration and nitrogenase activity from the two legumes were expressed conventionally either on a per plant or per gram nodule fresh weight basis, and where microscopy was done, on the basis of nitrogenase-containing, N(2)-fixing units (i.e. per bacteroid, per infected cell, or per gram infected tissue). In both legumes, acetylene reduction, N fixed and ureide content expressed on the basis of whole plants or per nitrogenase-containing units were very significantly correlated with values of leghaemoglobin concentrations expressed in a similar manner. The use of mathematical correlations in this study involving leghaemoglobin concentrations and various indices of N(2) fixation indicated a strong functional relationship between the two proteins in symbiotic legumes. These findings confirm previous suggestions that leghaemoglobin and the nitrogenase complex are two proteins closely associated with N(2)-fixing efficiency in legume root nodules.     

Growth and development of soyabean cv. TK5 as affected by tropical daylengths, day/night temperatures and nitrogen nutrition

Two experiments were done in Saxcil growth cabinets in order to investigate the effects of climatic factors and nitrogen nutrition on the growth, reproductive development and seed yield of soyabean cv. TK5. In the first, plants were grown to maturity in eight environments comprising all combinations of two short daylengths (11 h 40 min and 13 h 20 min), two day (27 and 33^oC) and two night (19 and 24^oC) temperatures. In the second, day temperature was kept at 33^oC but the night temperature was varied (19 and 24^oC) as was the mineral nitrogen supply (20 and 197 ppm N) to plants which were either inoculated or not with an effective single strain of Rhizobium. Taller, more branched, later flowering plants were produced in the longer daylength but seed yield was hardly affected because the components of yield did not all respond similarly. In the higher day temperature treatments seed yield per plant was reduced by half because all yield components were adversely affected - pods per plant by 34 %, mean seed dry weight by 24 % and seeds per pod just slightly. There was a marked effect of the higher night temperature which promoted early vegetative growth, induced early flowering and although the number of pods per plant was, overall, reduced by 48 %, seed yield per plant was little affected as mean seed dry weight was increased by 37 % and the number of seeds per pod was also increased slightly. Prior to flowering, nodulated plants obtained about two thirds of their total nitrogen requirement via direct uptake and one third through the symbiotic system. Vegetative dry weight and plant nitrogen content were increased by the higher mineral nitrogen level and, although height was slightly diminished, more branches were produced. Seed yield, however, was only slightly increased. These experiments have shown that night temperature is an environmental factor of major importance for the growth of this soyabean cultivar. They have provided, also, a more rational basis for interpreting seasonal variations in growth and seed yield of soyabean in the tropics where, clearly, day and night temperature effects can override those of daylength and nitrogen nutrition.     

Effects of Nitrogen Nutrition on Leaf Expansion and Photosynthesis of Trifolium subterraneum L. II. Comparison between Nodulated Plants and Plants Supplied with Combined Nitrogen

Subterranean clover plants depending on symbiotic nitrogen fixationhad smaller leaf areas than control plants supplied with combinednitrogen in the nutrient solutions. There were no differencesin chlorophyll content per unit fresh weight of leaves or petioles,nor in net rates of carbon dioxide uptake per unit leaf areaat light intensities above 2000 fc and at carbon dioxide concentrationsabove 300 ppm. Dark respiration by the shoots of the nodulatedplants was considerably higher than for the controls. This couldhave been a direct result of nodule activity and is suggestedas a possible factor contributing to the slower growth of theseplants compared with the controls. A comparison of the nitrogen contents of shoots and roots showeda sub-optimal nitrogen status, particularly in the roots, ofthe nodulated plants. This is suggested as another factor contributingto the slower growth of the nodulated plants compared with thecontrols. The response patterns before and after the addition of combinednitrogen differed in a number of important respects from thosefound previously under conditions of a sub-optimal nitrogensupply in the nutrient solution outside the roots. These arebriefly discussed.     

Studies on the nutrition of the forage legumes

Summary Cobalt has been shown to have a beneficial effect on the growth ofT. subterraneum, whether the plants are nodulated or not, and which is largely independent of the effectiveness of the rhizobial symbiont used.With effectively nodulated plants, an increase in the supply of combined nitrogen reduced both the nodulation (in grams per gram of root) and the haem content of the nodules. Within the ranges of concentration used, cobalt had little effect in counteracting this, which is in contrast with the results previously reported for the interaction of copper and nitrate supply.Part of the data on which this paper is based are from the thesis submitted by S.B.W. for the degree of Ph.D. of the University of Nottingham.     

In vivo nitrate reductase activities in different organs of lucerne plants: Effects of combined nitrogen during first vegetative growth and after shoot harvest

Nitrate reductase (EC 1.6.6.1–3; NR) activity was evaluated in nodulated lucerne ( Medicago sativa L. cv. Europe) grown aeroponically in both the presence and absence of applied nitrogen. Determination of in vivo NR activity was done with organ pieces in 0.1 M K⁺-phosphate, pH 7.5, 0.1 M KNO₃ and 1% n -propanol. NR activity was detected in all plant parts. Leaves accounted for 40% of the whole plant activity. Root activity was as high as leaf activity. Stem NR activity accounted for 14 to 20% of the total plant activity. NR activity was also detected in symbolically dependent plants grown without combined nitrogen. Nodule NR in symbolically dependent plants accounted for 17% of the tolal plant aclivity. When nitrate was present in the nulrienl medium, NR increased 5-fold as compared lo N₂-dependenl plants. Varying levels of nitrale (1.65 to 4 m M ) had no influence on leaf or stem activities. However, root NR activity seemed to be related to the nitrale concentration in the nulrient medium. Throughoul inilial vegelative growth, in vivo NR and nitrogenase (acelylene reduction) increased simultaneously. After shoot harvest, nitrogenase (acetylene reduction) aclivity drastically decreased with reduction of photosynthate supply, whereas NR increased in all organs, especially in N₂-dependenl plants.     

甘蔗—大豆间作对大豆鲜荚产量和农艺性状的影响

为探讨甘蔗-大豆间作模式对大豆鲜荚产量和农艺性状的影响,于2009—2011年连续3年在广州市华南农业大学农场进行大田试验,试验设置2种施氮水平(常规施氮(525kg·hm-2)和减量施氮(300kg·hm-2))和3种种植模式(甘蔗-大豆(1∶1)、甘蔗-大豆(1∶2)、单作大豆)。结果表明:甘蔗-大豆间作(1∶2)模式下,2009年减量施氮水平的大豆鲜荚产量较常规施氮水平提高了33%,2010和2011年不同施氮水平间均无显著差异;甘蔗-大豆间作模式对大豆的单株鲜荚重、多粒荚数和百粒鲜重无显著影响;大豆单株鲜荚重与多粒荚数在不同种植模式下均呈显著相关(P<0.05),在常规施氮间作模式下与大豆单株荚数呈显著相关(P<0.05)。甘蔗-大豆间作没有降低大豆的单株鲜荚产量,也没有对大豆的农艺性状产生负面影响,从增产增收、提高土地生产力来考虑,减量施氮模式下甘蔗-大豆间作具有一定的可行性。     

A Quantitative Study of Fixation and Transfer of Nitrogen in Alnus

A quantitative study of the fixation of nitrogen and of itstransfer from the nodules to the remainder of the plant hasbeen made in Alnus plants during their first season of growth.Fixation per plant reached a maximum in late August but fellrapidly with the onset of autumn, while fixation per unit dryweight of nodule tissues was greatest in young nodules and wasof the same order as in nodulated legumes. Throughout the growthseason there was a steady transfer from the nodules of some90 per cent, of the nitrogen fixed. The rate of fixation relativeto the growth of the endophyte is much higher than in free-livingnitrogen-fixing organisms and is clearly governed by the nitrogenrequirements of the entire symbiotic system. These findingsare compatible with the view that the fixation process is extracellularto the endophyte.     

Studies on Genetic Male-Sterile Soybeans: I. Distribution of Plant Carbohydrate and Nitrogen during Development

Soybeans (Glycine max [L.] Merr. cv. NC 69-2774) were used to study the nonstructural carbohydrate and nitrogen content of plant tissues, and nitrogenase activity throughout the development of male-sterile and male-fertile plants. Male-sterile plants set approximately 85% fewer pods plus seed than the male-fertile siblings and retained green leaves until a killing frost at 145 days after emergence. Reduced pod set caused increased carbohydrate accumulation in the leaf and root systems of male-sterile plants. Total carbohydrate in roots of male-sterile plants increased from 1.7 to 7.6 times that in the male-fertile roots. A high proportion (60 to 70%) of the male-sterile root carbohydrate was starch. Apparently, root starch was not metabolized by the male-sterile plants. Late in plant development per cent nitrogen was higher in the male-sterile soybean tissues. However, no difference was found in the ability of the nodulated root systems from either genotype to fix nitrogen.     

Some effects of environmental stress on seed yield of cowpea (Vigna unguiculata (L.) walp.) cv. Prima

Summary Separate experiments examined nodulation and seed yield of cowpea cv. Prima after (a) changes in the level of combined nitrogen from 25 to 0 or 60 ppm N, (b) cycles of wilting and rehydration, and (c) shading to ca 50% full daylight. Plants were grown in the simulated tropical environment of a plastics bubble house and experienced these changes over the growth stages: emergence to first flower, first flower to mid pod-fill or mid pod-fill to maturity.Seed yields of nodulated plants were unaffected by combined nitrogen supply and almost double those of non-nodulated plants (100 g cf 56 g per plant)-due mainly to increases in pod number per plant and mean seed weight. Reducing the nitrogen level from 25 to 0 ppm N, especially between mid pod-fill and maturity, reduced seed yields of non-nodulated plants to 36 g per plant. At the first flowering stage, plants grown without combined nitrogen had nitrogenase activities less than 10% of those supplied with 25 ppm N; 60 ppm N at any stage of development more than halved nitrogenase activity when compared with plants supplied with 25 ppm.Repeated wilting prior to flowering markedly reduced seed yields compared with the unstressed controls (41 g cf 76 g per plant)-mainly by decreasing subsequent pod production. Nodule weight and nitrogenase activity per plant were also much reduced. Wilting after flowering did not reduce yield, and nitrogenase activity was less affected.Shading throughout, or from first flower onwards, reduced seed yield by about 25% because fewer pods were produced. All shading treatments significantly increased mean seed weight compared with unshaded controls (116–121 mg cf 105 mg).One of a series of papers describing work undertaken in a collaborative project with the International Institute of Tropical Agriculture, Nigeria, sponsored by the U.K. Ministry of Overseas Development.Soil Microbiology Department, Rothamsted Experimental Station, Harpenden, Herts.Soil Microbiology Department, Rothamsted Experimental Station, Harpenden, Herts.     

Effect of nodulation on assimilate remobilization in soybean

The objectives of this work were to determine the effect of nodulation on dry matter, reduced-N, and phosphorus accumulation and partitioning in above-ground vegetative parts and pods of field-grown soybean (Glycine max [L.] Merr. cv Harosoy).

From comparison of nodulated and nonnodulated isolines, it was estimated that nodulated plants attained 81 and 71% of total-plant (above ground) N from uptake of soil N in 1981 and 1982, respectively. These data, along with visibly greener leaves of nodulated plants, led us to assume that nonnodulated plants were under a moderate N stress relative to nodulated plants. Nonnodulated plants accumulated less total-plant N and partitioned less dry matter and N to the pods, compared with nodulated plants. This occurred even though net photosynthesis, as estimated by rate and amount of dry matter accumulation, was the same for both nonnodulated and nodulated plants. Rate of dry matter and reduced-N accumulation in pods was less for nonnodulated than for nodulated plants while duration of podfill was similar for both isolines. From these data we concluded that moderate N stress affected partitioning of photosynthate rather than net photosynthesis, and that N played a role in translocation of photosynthate to the pods. Total plants (above-ground portion) and pods of both nodulated and nonnodulated plants accumulated similar amounts of phosphorus, which indicated that phosphorus and N accumulation were independent.

Remobilization of nitrogen and phosphorus from vegetation to pods preceded dry matter remobilization. It appeared that either more nitrogen accumulation prior to podfill, or continued nitrogen assimilation during podfill would increase nitrogen and dry matter partitioning to pods, but that increasing photosynthesis without concomitantly increasing nitrogen input may not necessarily result in enhanced seed production.

     

Comparisons of the Respiratory Effluxes of Nodules and Roots in Six Temperate Legumes

The specific respiration rates of nodulated root systems, ofnodules and of roots were determined during active nitrogenfixation in soya bean, navy bean, pea, lucerne, red clover andwhite clover, by measurements on whole plants before and afterthe removal of nodule populations. Similar measurements weremade on comparable populations of the six legumes, lacking nodulesbut receiving abundant nitrate-nitrogen, to determine the specificrespiration of their roots. All plants were grown in a controlled-environmentclimate which fostered rapid growth. The specific respiration rates of nodulated root systems ofthe three grain and three forage legumes during a 7–14-dayperiod of vegetative growth varied between 10 and 17 mg CO₂g^–1 (dry weight) h^–1. This mean value consistedof two components: a specific root respiration rate of 6–9mg CO₂ g^–1 h^–1 and a specific nodule respirationrate of 22–46 mg CO₂ g^–1 h^–1. Nodule respirationaccounted for 42–70 per cent of nodulated root respiration;nodule weight accounted for 12–40 per cent of nodulatedroot weight. The specific respiration rates of roots lackingnodules and utilizing nitrate nitrogen were generally 20–30per cent greater than the equivalent rates of roots from nodulatedplants. The measured respiratory effluxes are discussed in thecontext of nitrogen nitrogen fixation, nitrate assimilation. Glycine max, Phaseolus vulgaris, Pisum sativum, Medicago sativa, Trifolium pratense, Trifolium repens, soya bean, navy bean, pea, lucerne, red clover, white clover, nodule respiration, root respiration, fixation, nitrate assimilation     

Field experiments on nitrogen fixation by nodulated legumes

Summary Phosphate increased nitrogen uptake by lucerne appreciably on a saline soil. Nitrogenous fertiliser or inoculation with an effective strain ofRhizobium meliloti did not increase the yield significantly. In soils where indigenousRhizobium japonicum was absent inoculation increased soybean yields and the additional fixed nitrogen removed by soybeans amounted to 40 to 120 kg ha⁻¹. Gram and groundnut also responded to Rhizobium inoculation in field trials.     

Symbiotic N2 fixation activity in relation to C economy of Pisum sativum L. as a function of plant phenology

The relationships between symbiotic nitrogen fixation (SNF) activity and C fluxes were investigated in pea plants (Pisum sativum L. cv. Baccara) using simultaneous 13C and 15N labelling. Analysis of the dynamics of labelled CO2 efflux from the nodulated roots allowed the different components associated with SNF activity to be calculated, together with root and nodule synthetic and maintenance processes. The carbon costs for the synthesis of roots and nodules were similar and decreased with time. Carbon lost by turnover, associated with maintenance processes, decreased with time for nodules while it increased in the roots. Nodule turnover remained higher than root turnover until flowering. The effect of the N source on SNF was investigated using plants supplied with nitrate or plants only fixing N2. SNF per unit nodule biomass (nodule specific activity) was linearly related to the amount of carbon allocated to the nodulated roots regardless of the N source, with regression slopes decreasing across the growth cycle. These regression slopes permitted potential values of SNF specific activity to be defined. SNF activity decreased as the plants aged, presumably because of the combined effects of both increasing C costs of SNF (from 4.0 to 6.7 g C g-1 N) and the limitation of C supply to the nodules. SNF activity competed for C against synthesis and maintenance processes within the nodulated roots. Synthesis was the main limiting factor of SNF, but its importance decreased as the plant aged. At seed-filling, SNF was probably more limited by nodule age than by C supply to the nodulated roots.     

A comparison between the effect of fresh and dried organic materials added to soil on carbon and nitrogen mineralization

Summary Incubation experiments were carried out at 29°C in which fresh chopped, dried chopped, or dried and ground material of wheat plants,Polygonum nodosum, Senecio congestus (R. Br. DC.) and lucerne was mixed with a heavy calcareous loam. The C/N ratios of these materials were 45.9, 32.0, 19.3, and 12.6, respectively. At intervals of one or two weeks the content of mineral nitrogen in the treated and untreated soils was determined. In this way the immobilised or released nitrogen could be calculated. Parallel to this experiment the production of CO₂ in the soils treated in the same way was determined.The experiments have clearly shown that the mineralization of the carbon and nitrogen of incorporated organic materials is influenced not only by the N-content, the C/N ratio of the materials and the amount of the materials, but also depends on whether the plant materials are in fresh chopped, or dried chopped, or dried and ground condition.In most cases a retardation of the rate of nitrogen mineralization was found after drying the organic materials due to an increase of the C/N ratio of the water-soluble organic fraction by drying. The depressing effect of drying on the rate of nitrogen mineralization was increased by a mechanical breaking-up of the dried materials, presumably due to an increased surface of the non-soluble carbonaceous compounds.     

The Effect of Soil Water Regimes on Leaf Water Potential, Growth and Development of Soybeans

The growth and development of soybeans (Glycine max L. cv. Amsoy) was studied at soil matric potentials of ?0.1 to ?1.0 bars. Chlorophyll, photosynthesis, and leaf nitrogen per plant was greatest at ?4 bars leaf water potential. Leaf area, number of internodes, plant height and dry weight of vegetative parts declined as leaf water potential decreased from ?2 to ?19 bars. Nitrogen content and nitrate reductase activity per g fresh weight determined the percentage protein of individual seeds but nitrogen content and nitrate reductase activity per plant determined the amount of total seed protein. The protein synthesized in the seed changed little in amino acid composition with changes in leaf water potential. Leaf water potentials above or below ?4 bars decreased yield, total protein and total lipid but plants produced the largest percentage of individual seed protein at ?19 bars leaf water potential.     

The relative performance of white clover genotypes with rhizobial and mineral nitrogen in agar culture and in soil

Vegetative propagules of adult plants of white clover grown in agar culture show a better agreement with the performance of adult plants grown in soil than that previously reported of nodulated seedlings. This suggests that, whilst results obtained from the inoculation of seedlings can only be extrapolated to other situations with great caution, laboratory techniques which allow nitrogen fixation assessments to be made on adult plants may be useful in predicting performance under more natural conditions. The growth of vegetative propagules of mature plants dependent on rhizo-bium fixed nitrogen was also compared with propagules receiving abundant inorganic nitrogen. In agar culture there were no differences in yield between nitrogen sources, indicating that the symbiotic associations could fix sufficient nitrogen for growth under these relatively restrictive conditions. In soil, however, only the slower genotypes came near to providing sufficient nitrogen from symbiosis to sustain their potential yield as measured from their growth with inorganic nitrogen. The more vigorous the genotype the greater was the short-fall in nitrogen supplied by the rhizobium association. Joint regression analysis showed that any apparent interactions between plant genotypes and the source of nitrogen was a function of the genotypes' potential growth rate. The importance of quantifying nitrogen fixation in relation to the growth potential of the host genotype is emphasized, and the need, in legume breeding programmes, for simultaneous selection of host and rhizobium is discussed.     

Theory for growth of plants derived from the nitrogen productivity concept

A theory is developed on the assumption that growth of plants is determined by the current amount of nitrogen in the plants. The nitrogen-growth relation is formalized in the nitrogen productivity concept (amount of biomass produced per amount of nitrogen in the biomass and per unit of time), which is essentially a constant for a given species under fixed environmental conditions. A number of results follow for increases in whole plant biomass: (A) The relative growth rate is a linear function of the internal nitrogen concentration. (B) The maximal relative growth rate uniquely determines the scaling of the time axis. (C) Exponential growth is consistent only with stable internal nitrogen concentration. Dose-response curves expressed in reduced variables (the ratio between a variable and the same variable for a plant growing under optimal conditions) are universal, so that all species and all environmental conditions yield the same curve. This is confirmed by experimental data. The shape (linear, exponential, etc.) of the nitrogen uptake curve is the only parameter differentiating these universal curves. The Mitscherlich curve or variations of it can be fitted very closely to the derived dose-response curves, except under exponential growth. A conclusion drawn from the analysis is that the results of nutrition experiments cannot be properly interpreted unless the variation with time of the amount of nitrogen in the plant is known. The theory can be extended to more complex situations, for example, time-varying environmental conditions.