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     

Salt Tolerance in the Halophyte Suaeda maritima L. Dum. Growth, Ion and Water Relations and Gas Exchange in Response to Altered Salinity

Clipson, N. J. W. 1987. Salt tolerance in the halophyte Suaedamaritima L. Dum. Growth, ion and water relations and gas exchangein response to altered salinity.—J. exp. Bot. 38: 1996–2004. Shoot and root fresh and dry weights and shoot sodium, chlorideand potassium contents were measured and shoot relative growthrates calculated in seedlings of Suaeda maritima over a periodof 11 d following a raising of culture solution salinity from0 to 200 mol m₃– NaCl. Growth, growth rates and sodiumand chloride contents, as compared to plants growing in theabsence of salt were increased whilst potassium contents declined.Shoot sodium accumulation rate and the rate of transport ofsodium from root to shoot, osmotic potential, and rates of photosynthesisand transpiration were also measured for up to 72 h after transferof plants originally growing at 0 and 200 mol₃– NaCl to200 and 400 mol m₃– NaCl respectively. Ion uptake andtransport rates were maximal 6-12 h after transfer and thendeclined to new steady-state levels within 48 h; osmotic potentialswere lowered over a 72 h period on average by approximately1·0 MPa; and after 9 h photosynthetic and transpirationrates were reduced by about 20percnt; and 30% respectively.Results are discussed in terms of the ability of halophytesto adjust to fluctuating salinity and to salt tolerance mechanismsin general. Key words: Suaeda maritima, salinity, gas exchange, growth, ion and water relations     

Effects of NaCl on Growth, Nitrogen Incorporation and Chemical Composition of Inoculated and NH4NO3 Fertilized Vicia faba (L. ) Plants

Vicia faba cv. Maris Bead was grown either on fixed nitrogenor on ammonium nitrate. After 4 weeks growth, nutrient solutionswere supplemented with 50, 75 and 100 mol m^–3 NaCl for15 d. Five harvests were made at weekly intervals, beginningat 4 weeks. Effects of salinity were directly related to dose,plant growth (fresh and dry weight) being depressed in bothN-fixing and N-fertilized plants. The number of nodules perplant and the proportion of those formed which developed intothe active nitrogen fixing state were depressed by salt stress.Increased size of nodules in salt-stressed plants only partlycompensated for the lower specific nitrogenase activity. Theeffects of salinity on plant nitrogen content were more pronouncedon N-flxing than on N-fertilized plants. The former took upmore Na⁺ and Cl^– than the latter: the implications ofthis and of ionic imbalance are discussed. Key words: Vicia faba, Growth, Salt stress, Nodulation     

Use of the Pressure Bomb for Hydraulic Conductance Studies

Stem sections of alfalfa (Medicago sativa L.) were subjectedto forced flows of both water and high molecular weight dextransolutions. In either case, stems responded in accordance withthe Ohm's law analogy for flow, i.e., flow was a linear functionof pressure, at pressures ranging from 7.0–500 kPa andvacuum from 25.69 kPa. Since conductance values were the samefor both pressure and vacuum powered flows, it is possible toextrapolate conductance values determined with a pressure bombto negative pressure gradients experienced by plants. Key words: Pressure bomb, Conductance, Medicago sativa     

The Effect of Irradiance on the Recovery of Soybean Nodules from Sodium Chloride-Induced Senescence

Soybean (Glycine max L. Merr) cv. Clarke plants inoculated withBradyrhizobium japonicum strain RCR3407 were grown either ina greenhouse with a low irradiance (200–400)µmolm^–2 s^–1) or in a controlled-environment growth cabinetwith a higher irradiance (600 µimol m^–2 s^–1).At 42 d plants were given a nitrogen-free nutrient solutioncontaining 50 mol m^–3 sodium chloride for 2 weeks andthen allowed to recover from salt-stress for a further 2 weeks. Salt treatment reduced plant growth by at least half in bothgrowth regimes, however, the controlled environment-grown (CEG)plants were five times larger than the greenhouse-grown (GG)plants in terms of dry weight and number/weight of nodules perplant, regardless of treatment. The structure of nodules, from both growth regimes, harvestedat the end of the 2 week salt-stress was similar to unstressedcontrol nodules. However, nodules harvested 1 week later fromboth CEG and GG plants had structural changes including degradationof bacteria in vacuoles around host cell nuclei, particularlyin the outer cell layers of the infected tissue. In addition,meristematic activity was seen in the cortex of some nodulesfrom GG plants. Young cells here contained infection threadsand newly-released bacteria. Nodules harvested 2 weeks after removal of the salt-stress fromCEG plants showed an apparent recovery from the stress. However,there was a very marked increase in the amount of starch inthe cortex which was not seen in equivalent GG nodules. In contrast,nodules from GG plants contained many vacuolate infected cellsand, consequently, a lowered bacteroid population. Further,meristematic activity was seen in a zone concentric to the infectedzone, newly-formed cells contained many large infection threadsand were interspersed with intercellular bacteria. The meristematicactivity increased the relative volume of cortical to infectedcells in these nodules. Growth conditions did not affect control nodule specific nitrogenaseactivity or oxygen diffusion resistance (R) and these parameterswere also not altered in CEG nodules exposed to salt plus the14 d recovery period. However, nitrogenase activity was greatlyreduced, and R increased by more than eight times in equivalentGG nodules exposed to salt plus recovery. It is hypothesized that the gross morphological changes werean attempt to counter salt toxicity and/or oxygen damage underconditions of reduced photosynthate supply to the nodules dueto the poor light levels in the greenhouse. However, soybeannodules supplied with adequate photosynthate were able to withstandand recover from long-term salt-stress with little alterationto their structural integrity. Key words: Soybean, sodium chloride, nitrogen fixation, light intensity, oxygen diffusion resistance     

Agrobacterium rhizogenes Mediated Transformation of the Forage Legumes Medicago sativa and Onobrychis viciifolia

Three cultivars of M. sativa and one cultivar of O. viciifoliawere evaluated for their response to inoculation with A. rhizogenesstrain A4T (containing pRiA4b). A cultivar-dependent responsewas observed in M. sativa with 94%, 25%, and 4% of infectedstem explants producing transformed roots in the cultivars Vertus,Regen-S, and Rangelander, respectively. In O. viciifolia cv.Hampshire Giant, an explant-dependent response was observedwith 78% and 50% of seedling cotyledon and hypocotyl explantsresponding, respectively. Leaf explants failed to produce transformedroots. Transformed roots showed plagiotropic and negativelygeotropic growth on hormone-free agar MS medium. Productionof transgenic shoots from O. viciifolia root cultures occurredspontaneously. Recovery of transgenic plants from M. salivacv. Rangelander was achieved by transfer of callus (inducedon UM medium containing 2·0mg dm^–3 2,4-D and 0·25mg dm^–3 kinetin) to MS medium containing 0·5 ingdm^–3 BAP and 0·05 mg dm^–3 NAA. Cultured rootsof both species synthesized opines (agropine and mannopine).Extensive morphological variation was observed in plants ofM. sativa (clone Al) and O. viciifolia (clone A4Tl) establishedin the glasshouse. DNA sequences homologous to TL-DNA and TR-DNAwere present in root clones and regenerated plants. Key words: Agrobacterium rhizogenes, Medicago sativa, Onobrychis viciifolia, transformed roots, transgenic plants     

Inhibition of Somatic Embryogenesis in Tissue 2Medicago sativa by Aminoethoxyvinylglycine, Amino-oxyacetic acid, 2, 4-Dinitrophenol and Salicylic Acid at Concentrations which do not Inhibit Ethylene Biosynthesis and Growth

Meijer, E. G. M. and Brown, D. C. W. 1988. Inhibition of somaticembryogenesis in tissue cultures of Medicago sativa by aminoethoxyvinylglycine,amino-oxyacetic acid, 2, 4-dinitrophenol and salicylic acidat concentrations which do not inhibit ethylene biosynthesisand growth. J. exp. Bot. 39: 263–270. The effects of aminoethoxyvinyglycine (AVG), amino-oxyaceticacid (AOA), 2, 4-dinitrophenol (DNP) and salicylic acid (SA)on ethylene production, tissue proliferation and somatic embryo-genesisin a recently developed rapid in vitro regeneration system ofMedicago sativa L. were examined. Contrary to numerous publications,AVG, AOA and DNP did not affect the rate of ethylene biosynthesis,while SA even caused an increase in ethylene production. Allfour compounds were, however, potent inhibitors of somatic embryoformation in the M. sativa tissue cultures, even at concentrationswhich did not affect tissue growth. Generally, a 5-d exposureto the inhibitors reduced the number and quality of somaticembryos obtained. It is suggested that the inhibitors may notreach the site of action of enzymes involved in ethylene biosynthesisand may possibly block other biosynthetic pathways which areof crucial importance to somatic embryo development. The resultsindicate that a delicate differentiation process like somaticembryogenesis is very sensitive to metabolic perturbances. Theresults are also discussed in the light of other known effectsof these four compounds on higher plants. Key words: Ethylene, Medicago sativa, somatic embryogenesis, tissue culture     

Salt Tolerance in the Triticeae: Growth and Solute Accumulation in Leaves of Thinopyrum bessarabicum

Gorham, J., McDonnell, E., Budrewicz, E. and Wyn Jones, R. G.1985. Salt tolerance in the Triticeae: growth and solute accumulationin leaves of Thinopyrum bessarabicum.—J. exp. Bot. 36:1021–1031. The diploid wheatgrass Thinopyrum bessarabicum was found towithstand prolonged exposure to 350 mol m^–3 NaCl in hydroponicculture. During the gradual addition of salt to the externalmedium, osmotic adjustment was rapidly achieved by the accumulationof Na and Cl. Following osmotic adjustment constant leaf Naand Cl concentrations were maintained, and K was retained ata high level. Thinopyrum bessarabicum may be described as anosmoconformer, adjusting its internal osmotic pressure to 400–500mOsmol kg^–1 above that of the external medium in hydroponicculture. Both slower shoot initiation and reduced leaf lengthcontributed to the reduced growth rates at higher salinities.Leaf width was not affected. Increasing salinity resulted inincreases in leaf concentrations of phosphate, glycinebetaine,sucrose and proline, and in decreases in the concentrationsof nitrate, sulphate, magnesium, calcium, total amino acidsand organic acids. Thinopyrum bessarabicum exhibits salt tolerancecharacters which may be useful in wheat breeding. Key words: Salt stress, solute accumulation, osmotic adjustment, Thinopyrum     

In Vitro Propagation of Potato (Solanum tuberosum L.)

Potato shoots were propagated in vitro by placing nodes fromsprouted tubers on Murashige and Skoog type medium without hormones.The vigour of growth and the rate of node production increasedwith both day-length and temperature over the ranges 8–24h and 15–25 °C respectively. Propagation rates ofup to x 10 per month were obtained. In vitro plantlets spontaneouslyformed roots either in agar or liquid cultures. Plantlets leftin the culture jars for 3–4 months eventually senescedand formed small tubers in 16 and 24 h day-lengths. In a day-lengthof 8 h vegetative growth continued by branching and no tuberswere formed. Solanum tuberosum L., potato, tissue culture, propagation, temperature, day-length     

The Effect of Water Stress on the Vegetative Growth of White Clover (Trifolium repens L): Comparison of Long-term Water Deficit and a Short-term Developing Water Stress

White clover plants were subjected to either a short-term developingwater stress or long-term stable levels of water deficit on‘water stress columns’. The short-term stress reducedplant water status to –2?0 MPa over 15 d. The water stresscolumns imposed only mild levels of water stress (a reductionof 0?35 MPa in leaf water potential for the more severe treatment)but these were maintained for several weeks. The absolute growthof plants on the control columns was maintained throughout theexperimental period. Vegetative growth was measured. Stolon, petiole, and laminagrowth were all reduced to some extent when plants were grownsymbiotically. The two regimes gave comparable results. Whennitrate was supplied there was no effect of water stress. Aconsiderably reduced absolute growth rate did not result ina similar decrease in final organ size. Stolon growth was mostreduced by water stress. Leaf death during water stress wasas important as changes in growth in determining final dry matteryield. Consequently, the yield of petiole and lamina from plantsgrown without supplied nitrate on the water stress columns waslower than that of stolon at the end of the treatment period. The merits of the water stress column system for imposing long-termwater deficit are discussed. Key words: Trifolium repens, white clover, water stress, vegetative growth     

The Effects of Nitrogen Supply on the Absorption and Distribution of Copper in Red Clover (Trifolium pratense L.) Grown in Flowing Solution Culture with a Low, Maintained Concentration of Copper

The absorption and distribution of Cu in red clover (Trifoliumpratense L.) were measured in plants grown in flowing solutionculture with Cu maintained throughout at 0.5 µg 1^–1and N supplied either as nitrate or through symbiotic fixation.Although there was a decrease in Cu absorption, both with time,and with a depleted nitrate supply, it increased to its formerrate when nitrate was adjusted to 10 mg N 1^–1 after aperiod of depletion. Differences in absorption between plantsdependent upon fixation and those supplied with nitrate wererelated to the slower initial growth of the plants fixing N.Considerable proportions (> 30 per cent) of the absorbedCu were retained by the roots. At the final harvest, and withthe exception of plants grown with nitrate adjusted to 0.1 mgN 1^–1 after a period of depletion, the proportion of theCu retained was related to the concentration of N in the roots.The different N treatments produced differences in Cu concentrationin the shoots, and the effects were greater in the youngestfully expanded leaves than in older leaves. Trifolium pratense L., red clover, absorption, copper, flowing solution culture, nitrogen     

Changes in the Total Alkaloid Content of Datura innoxia Mill. Subjected to Salt Stress

Datura plants were grown on a clayed support and subjected tosalinity stress (153.8 mol m^–3 NaCl) at the 6-leaf stage.Salt treatment increased total alkaloid content in young leaves.The results indicated that at the organ level tropane alkaloidaccumulation was related to plant growth. Key words: Datura innoxia, salt stress, tropane alkaloids     

Growth of Lucerne (Medicago sativa L.) Exposed to Sulphur Dioxide

The effect of exposure to sulphur dioxide (0 or 96 µgm^–3 SO₂) on the growth and sulphur content of lucerne(Medicago sativa L.) was examined in a period of 135 d duringwhich the plants were harvested four times. The lucerne wasgrown in pots of soil with and without the addition of sulphateand of nitrate. Evidence of sulphur deficiency, including areduction in the weight, the number and the sulphur contentof shoots, was found in plants grown without added sulphate.Deficiency was alleviated through exposure of plants to SO₂.Apart from reducing shoot weight at one harvest and generallyincreasimg the concentration of sulphur in the shoots, exposureto SO₂ had no significant effect on plants grown with addedsulphate. The yield of shoots was greater, and was reduced toa lesser extent with sulphur deficiency, in plants grown withadded nitrate than in those dependent on rhizobia. Whilst thetranspiration coefficient increased greatly in sulphur-deficientplants, it was reduced where otherwise similarly treated plantswere exposed to SO₂; this treatment did not alter the coefficientin plants with an adequate supply of sulphur from the soil.     

Influence of Osmotic Stress on Nitrate Reductase Activity in Wheat (Triticum aestivum L.) and the Role of Abscisic Acid

Wheat seedlings (Triticum aestivum L. cv. Timmo) were treatedwith up to 20% (w/v) polyethylene glycol (PEG, mol. wt. 3350)in the nutrient medium for 6 d. Shoot growth and nitrate transportand metabolism were substantially affected by PEG treatment.At 20% PEG (corresponding to a water potential of approximately–1.6 MPa), which caused plants to wilt within 1–2h, activity of nitrate reductase (NR) declined with a half-lifeof approximately 5 h in both roots and shoots. The decline wasconsiderably slower at lower PEG concentrations. Significantincreases in levels of abscisic acid (ABA) only occurred inshoots. Application of ABA to intact plants or excised shootsdid not induce or accelerate decline in shoot NR activity. Therapid decline in NR activity during wilting appears unrelatedto both nitrate flux and ABA. At lower PEG concentrations andin the long-term, however, NR activity corroborates rates ofboth transport and growth-related utilization of nitrate. Therole of ABA in this context appears to be indirect through itsaction on stomatal function which reduces water flux and gasexchange. Key words: Stress, nitrate reductase, abscisic acid (ABA)     

The Uptake of Phosphate by Plants from Flowing Nutrient Solution: IV. EFFECT OF PHOSPHATE CONCENTRATION ON THE GROWTH OF TRIFOLIUM REPENS L. SUPPLIED WITH NITRATE, OR DEPENDENT UPON SYMBIOTICALLY FIXED NITROGEN

Breeze, V. G. and Hopper, M. J. 1987. The uptake of phosphateby plants from flowing nutrient solution. IV. Effect of phosphateconcentration on the growth of Trifolium repens L. suppliedwith nitrate, or dependent upon symbiotically fixed nitrogen.—J.exp. Bot. 38: 618–630. Nodulated white clover plants were subjected to a range of phosphateconcentrations in flowing solution culture (0.32 to 8.0 mmolm^–3 P) at 41 d from sowing, either supplied with nitrateor dependent on symbiotically-fixed nitrogen. No effect of phosphateconcentration in solution on dry matter production, relativegrowth rate, root/shoot ratio, or water soluble carbohydrateconcentration of the plant tissue was observed after 24 d fromthe start of the experiment, although the plants supplied withnitrate yielded more than the others. Phosphate uptake throughoutthe experimental period was related to the solution concentration,but the source of nitrogen did not affect the phosphorus concentrationsof the shoots. However, the roots of the plants dependent onsymbiotically-fixed nitrogen had higher concentrations of phosphorusthan those supplied with nitrate, but this did not appear tobe due to an increased phosphorus requirement for nitrogen fixation,because the amount fixed was unaffected by the phosphate concentrationin solution. The cation-anion balance showed that plants dependenton nitrogen fixation had no larger requirement for calcium thanplants supplied with nitrate, but a requirement for hydroxylions equivalent to over 130 kg lime per tonne of dry shoot.It is suggested that the enhanced phosphate uptake by plantsdependent on nitrogen fixation is due to this need for a cation-chargebalancing anion. Key words: Phosphate uptake, nitrogen fixation, Trifolium repens L., repens L., cation-anion balance, flowing solution culture     

Nitrate Reductase Acivity in Leaves of Barley (Hordeum vulgare) and Durum Wheat (Triticum durum) during Field and Rapidly Applied Water Deficits

Smirnoff, N., Winslow, M. D. and Stewart, G. R. 1985. Nitratereductase activity in leaves of barley (Hordeum vulgare) anddurum wheat (Triticum durum) during field and rapidly appliedwater deficits.-J. exp. Bot 36: 1200-1208. The effect of field and rapidly applied water deficits on nitratereductase activity in the leaves of two barley varieties andone durum wheat variety was investigated. In field experimentsplants were subjected to irrigation at different rates in threeMediterranean environments by means of a line source sprinklerirrigation system. The environments differed in rainfall andnitrogen fertility. Plant water potentials decreased from –1.5MPa to between –2.5 and –3.0 MPa as the irrigationrate decreased. Nitrate reductase activity in the leaves ofthese plants during heading was either unaffected or sometimesincreased where the least water was supplied. Nitrate reductaseactivity was highest in the plants growing with an ample nitrogensupply irrespective of water regime. In contrast, seedlingssubject to rapidly applied water stress over 6 d lost 30-85%of their nitrate reductase activity when leaf water potentialfell from between –0.33 and –0.75 MPa to between–O.93 and –2.04 MPa. The decrease was less in theyoung leaves than in the old leaves. Polyethylene glycol inducedosmotic stress resulted in a drop in leaf water potential from–0.20 MPa to between –1.05 and –1.20 MPa alongwith a loss of 40-85% of leaf nitrate reductase activity after48 h. It is suggested that maintenance of nitrate reductase activityin field grown barley and durum wheat plants reflects an acclimationto water deficit Maintenance of nitrate assimilation duringwater stress may allow continued synthesis of nitrogenous compatiblesolutes using the excess photochemical energy available duringstomatal closure. Key words: Nitrate reductase, water stress, barley, durum wheat     

Carbon and Nitrogen Assimilation by Vicia faba L. at Low Temperature: the Importance of Concentration and Form of Applied-N

Low temperature (6 C) growth was examined in two cultivarsof Vicia faba L. supplied with 4 and 20 mol m^–3 N as nitrateor urea. Both cultivars showed similar growth responses to increasedapplied-N concentration regardless of N-form. Total leaf areaincreased, as did root, stem and leaf dry weight, total carboncontent and total nitrogen content. In contrast to findingsat higher growth temperatures, 20 mol m^–3 urea-N gavesubstantially greater growth (all parameters measured) than20 mol m^–3 nitrate-N. The increased carbon content per plant associated with increasedapplied nitrate or urea concentration, or with urea in comparisonto nitrate, was due to a greater leaf area per plant for CO₂uptake and not an increased CO₂, uptake per unit area, carbon,chlorophyll or dry weight, all of which either remained constantor decreased. Nitrate reductase activity was substantial inplants given nitrate but negligible in plants given urea. Neitherfree nitrate nor free urea contributed greatly to nitrogen levelsin plant tissues. It is concluded that there is no evidence for a restrictionin nitrate reduction at 6 C, and it is likely that urea givesgreater growth than nitrate because of greater rates of uptake. Vicia faba, broad bean, low temperature growth, carbon assimilation, nitrogen assimilation     

Triadimenol Increases Nitrate Levels and Nitrate Reductase Activity in Canola Leaves

Nitrate reductase activity in the first true leaves of canola(Brassica napus L.) seedlings grown in one-quarter strengthHoagland's solution from seeds pretreated with triadimenol (0.3or 30 g (a.i.) kg^–1 of seed) was higher than controlsduring the growth period of 15 to 25 d after planting. Triadimenolalso increased chlorophyll levels, the increase being more pronouncedat its lower concentration. The treatment also increased theweight and nitrate content of the leaves. When seedlings weregrown in nutrient solution containing 1 to 20 mM nitrate, theincrease in nitrate reductase activity by triadimenol was higherat lower rather than at higher nitrate concentrations. The nitratelevels and Kjeldahl nitrogen in the triadimenol-treated leaveswas higher than the controls at concentrations of added nitrateabove 2 mM. Addition of nitrate to plants grown in ammonium,increased nitrate reductase activity more in plants grown fromtriadimenol-treated seeds than controls. However, addition of10µM triadimenol for 24 h to ammonium-grown plants hadlittle effect on enzyme activity, both in the absence as wellas the presence of nitrate. This study demonstrates that triadimenolincreases nitrate reductase activity and nitrate accumulationin the leaves and at least part of the increased enzyme activityis independent of nitrate accumulation. Key words: Triazoles, nitrate content, nitrate reductase activity     

Studies on the Physiology of Nodule Formation: With two Figures in the Text: A Reappraisal of the Effect of Preplanting

Nutman (Arm. Bot. 21, 321, 1957) found that preplanting agarslopes with Trifolium pratense L. or Medicago sativa L. advancedthe time when second lots of plants of these species grown onthe same slopes initiated nodules, and depressed the total numberof nodules formed. He attributed these effects to root secretionswhich at low concentration hasten initial nodulation but athigher concentrations inhibit nodule formation. Further workhas now shown that initial nodulation is stimulated becausethe preplant removes traces of nitrate from the medium. Theamount of nitrate in the tap water used to prepare the medium(6?5 p.p.m. N) also increases the number of nodules formed onthe control plants, and this effect explains to a considerableextent the depression of nodule numbers by preplanting. Initial nodulation was delayed by small amounts of nitrate andnitrite but not by other forms of combined nitrogen (ammonium,asparagine, and urea). All forms of combined nitrogen testedincreased the number of nodules formed over a period of 8 weekswhen supplied at an initial concentration of 20 p.p.m. N.