Molecular and morphological examination of Cyrtobagous sp. collected from Argentina, Paraguay, Brazil, Australia, and Florida

Two members of the floating fern genus Salvinia (Salviniaceae), S. minima Baker and S. molesta Mitchell, have established in the United States. Cyrtobagous salviniae Calder and Sands (Coleoptera: Curculionidae), long established on Florida S. minima, was released in Texas and Louisiana as a biocontrol agent for both species. Subsequently, sequence analysis of the 28S rRNA D2 expansion domain suggested that the Florida and Brazilian populations (used worldwide for biocontrol) of C. salviniae might constitute two cryptic species. In response, the Brazilian weevil was imported from Australia and released instead onto S. molesta. We sampled C. singularis Hustache and C. salviniae from their native ranges in Brazil, Argentina, and Paraguay and sequenced them (D2) along with Australian and Florida samples. The genetic distance between C. singularis and C. salviniae samples is much greater (almost 5×) than the distance between either the Florida and Brazilian samples or the Brazilian and Argentinean/ Paraguayan C. salviniae samples. Since C. singularis and C. salviniae are cryptic species, the Florida and Brazilian populations (or for that matter Brazilian and Argentinean/Paraguayan) could reasonably be described as demes or ecotypes. Occurrence data indicates that, in parts of their ranges, C. salviniae and C. singularis are not only sympatric but also feed on the same plant species at the same site. While host adaptation (species preferences) likely occurs within local demes, both species seem capable of adapting to the available resource (Salvinia species). Finally, a polymerase chain reaction (PCR) primer was developed to distinguish the Florida and Brazilian/Australian types.     

Asymmetric impacts of two herbivore ecotypes on similar host plants

1. Ecotypes may arise following allopatric separation from source populations. The simultaneous transfer of an exotic plant to a novel environment, along with its stenophagous herbivore, may complicate more traditional patterns of divergence from the plant and insect source populations. 2. The present study evaluated herbivory effects by two Cyrtobagous salviniae ecotypes on two species of Salvinia: the larger S. molesta and the smaller S. minima. Evaluations were based on relative growth rates, biomass production, coverage, and nutrient cycling in a series of complementary and comparative laboratory and outdoor tank experiments. 3. In general, the experiment results indicated that the smaller Florida ecotype of C. salviniae impacted both Salvinia species more than the larger Brazil ecotype. Herbivory, especially by the Florida ecotype, also improved water quality and accelerated nutrient cycling by increasing the rates of litterfall from the standing crops of salvinia. 4. The smaller size of the Florida ecotype may be adaptive by allowing maximal exploitation of host plants via internal larval feeding, which presumably reduces predation risk while increasing damage to the plant. These findings provide support for relying on the suppressive effects of the Florida ecotype in Florida, rather than introducing the Brazil ecotype to counter new infestations of S. molesta.     

Compensatory branching and changes in nitrogen content in the aquatic weed Salvinia molesta in response to disbudding

Summary Secondary side branching in Salvinia molesta plants grown in nutrient solution was dependent on the availability of nitrogen. The compensatory response of S. molesta to damage by complete manual disbudding was the production of extensive secondary side branching at nitrogen levels below those required for this type of branching in undamaged plants. After 28 days damaged plants had the same number of ramets as undamaged plants but the dry weight was considerably reduced.After 7 days there was no change in dry weight with partial or complete disbudding. However completely disbudded plants had no new ramets and plants with all terminal buds removed had numbers of ramets significantly reduced. The concentration of nitrogen in the tissue of new growth was higher in damaged than undamaged plants. The results are discussed in relation to the biological control of S. molesta by the bud-eating weevil, Cyrtobagous salviniae.     

Suppression of Salvinia molesta Mitchell in Texas and Louisiana by Cyrtobagous salviniae Calder and Sands

Release and evaluation studies of the Brazil population of Cyrtobagous salviniae on Salvinia molesta were conducted originally at 18 sites in Texas and Louisiana from 1999 through 2005. However, overall project results could only be continually evaluated at two release and two control sites because the remainder were eventually destroyed or corrupted by floods, droughts, or herbicides. Mean fresh weight biomass of S. molesta ranged from 15.5 kg FW m⁻² during the summer to as low as 2.1 kg FW m⁻² during the winter prior to the release of C. salviniae. Insect populations established within a year of release and initially spread slowly. Damage to S. molesta increased with increasing C. salviniae detections while S. molesta biomass and surface coverage declined at both release sites by more than 99% while remaining unchanged at the control sites. Water in release sites registered higher levels of dissolved oxygen, higher temperatures, and higher pH than water in control sites. This study provides another example of the effectiveness of C. salviniae against S. molesta even in more temperate climates.     

A comparative study on the intrinsic rates of increase ofCyrtobagous singularis andC. salviniae on the water weedSalvinia molesta

The intrinsic rates of increase (r_m) ofCyrtobagous salviniae Calder & Sands from Brazil andC. singularis Hustache from Trinidad W.I., were determined in the laboratory at 23°C, 27°C and 31°C on nitrogen-rich plants of the aquatic weed,Salvinia molesta Mitchell.Variation in oviposition and immature survivorship accounted for most of the differences between species in r_m values (exponential growth of a stable-age population in a non-limiting environment). Values for r_m were higher forC. salviniae (0.210, 0.366, 0.404) than forC. singularis (0.148, 0.140, 0.064) at the three temperatures respectively. At all temperatures,C. salviniae laid seven times more eggs thanC. singularis while at 31°C oviposition was reduced for both species by 45%, and was accompanied by a reduction in egg hatch. Oviposition byC. salviniae was almost continuous (92% of weeks with some eggs laid) whereas oviposition byC. singularis was intermittent (50% of weeks) with intervals averaging 3 weeks without oviposition. Nitrogen concentration inS. molesta affected reproduction byC. singularis more thanC. salviniae, an increase of 0.1% (dry wt) increasing weekly oviposition by 7.0% and 3.6% respectively.The differences in r_m for the two weevil species are discussed in relationship to their potential as biological control agents.

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Taux intrinsèques d'accroissement deCyrtobagous singularies Hustache andC. salviniae Calder & Sands, utilsés dans la lutte biologique contreSalvinia molesta Mitchell

Résumé Les taux intrinsèques d'accroissement (r_m) deC. salviniae du Bresil et deC. singularis de Trinidad, ont été établis au laboratoire à 23°C; 27°C; 31°C, sur des plants deS. molesta riches en azote.Les différences entre les r_m des deux espèces provenaient pour l'essentiel de la ponte et de la mortalité préimaginale. Aux 3 températures, les valeurs de r_m deC. salviniae (0.210; 0.366; 0.404) étaient supérieures à celles deC. singularis (0.148; 0.140; 0.064). A toutes les températuresC. salviniae a pondu 7 fois plus d'oeufs queC. salviniae; à 31°C, la ponte a été réduite de 45% pour les 2 espèces et été accompagnée d'une diminution des taux d'éclosion. La ponte deC. salvinae était presque continue (92% des semaines avec des oeufs), tandis que celle deC. singularis était intermittent (50% des semaines avec pontes), les interruptions étant en moyenne de 2 semaines. La teneur deS. molesta en azote a affecté la reproduction deC. singularis plus que celle deC. salviniae; un accroissement de 0.1% en poids sec, augmentant les pontes hebdomadaires respectivement de 7% et de 3.6%.Les différences de valeur de r_m des 2 espèces sont examinées pour évaluer leurs potentialités comme éléments de la lutte biologique.

     

Pre-incubation of Bradyrhizobium japonicum with jasmonates accelerates nodulation and nitrogen fixation in soybean (Glycine max) at optimal and suboptimal root zone temperatures

Jasmonic acid (JA) and methyl jasmonate, collectively known as jasmonates, are naturally occurring in plants; they are important signal molecules involved in induced disease resistance and mediate many physiological activities in plants. We studied the effect of JA and its methyl ester, methyl jasmonate (MeJA), on the induction of nod genes in Bradyrhizobium japonicum GG4 (USDA3) carrying a plasmid with a translational fusion between B. japonicum nodY and lacZ of Escherichia coli, and the expression activity was measured by β-galactosidase activity. Both JA and MeJA strongly induced the expression of nod genes. They have little or no deleterious effects on the growth of B. japonicum cells, while genistein (Gen) showed inhibitory effects. We further studied the effect of JA- and MeJA-induced B. japonicum on soybean nodulation and nitrogen fixation under optimal (25°C) and suboptimal (17°C) root zone temperature (RZT) conditions. B. japonicum cells were grown in liquid yeast extract mannitol media and induced with a range of Gen, JA, and MeJA concentrations, including a treatment control with no inducer added. Soybean seedlings were grown at 25 or 17°C RZT with a constant air temperature (25°C) and inoculated, at the vegetative cotyledonary stage, with various B. japonicum induction treatments. Addition of Gen or jasmonates to B. japonicum, prior to inoculation, enhanced nodulation, nitrogen fixation, and plant growth at suboptimal RZT conditions. A higher concentration of Gen was inhibitory at 25°C, while this same concentration was stimulatory at 17°C. Interestingly, pre-incubation of B. japonicum with JA and MeJA enhanced soybean nodulation and nitrogen fixation under both optimal and suboptimal RZTs. We show that jasmonates are thus a new class of signaling molecules in the B. japonicum-soybean symbiosis and that pre-induction of B. japonicum with jasmonates can be used to enhance soybean nodulation, nitrogen fixation, and early plant growth.     

Biological control of giant salvinia (Salvinia molesta) in a temperate region: cold tolerance and low temperature oviposition of Cyrtobagous salviniae

Success of Cyrtobagous salviniae Calder & Sands (Coleoptera: Curculionidae) for biological control of Salvinia molesta D. S. Mitchell in temperate regions has been less reliable than in tropical and subtropical regions and this difference is presumed to be due to greater winter mortality. We measured the cold tolerance of C. salviniae by comparing chill coma recovery time and survival of adults after exposure to freezing conditions among four geographic populations collected from Florida, Louisiana, Texas and Australia. Effects of winter temperature acclimation on low temperature oviposition also were determined. The Australian population was more cold tolerant than the three US populations. No oviposition by C. salviniae was observed at a water temperature of 17 °C and the oviposition rate at 19 °C was less than at 21, 23 and 25 °C. We suggest that introduction of cold tolerant strains of C. salviniae could increase its effectiveness in temperate regions of the US.     

Biological control of Salvinia molesta in some areas of Moremi Game Reserve,Botswana

The Curculionid weevil, Cyrtobagous salviniae Calder & Sands, has been established on Salvinia molesta (salvinia) in Botswana. The study determined the intensity of weevil activity on the weed at the margin and the centre of four selected sites on the Khwai system of the Okavango Delta. Random samples of salvinia were collected from each site at monthly intervals in 1998 to extract weevils and to demonstrate the effect of weevil on the weed. The rate at which the weed was controlled at different sites varied with mat and weevil density. The biological control at Paradise Pools was moderate without much increase in the weed biomass per m², while fluctuations in both weevil and plant populations were noticed in the Khwai streams. Weevil numbers remained low at Dombo for the first nine months of the year, during which time the mat density increased. With the onset of higher temperatures, weevil numbers increased to 155 and 457/kg fresh weight of salvinia at the margin and centre respectively in Dombo Pool in early summer. A significant control in MGR 6 HATAB pool between mat biomass and the weevil number resulted in the disappearance of the weed in three months. This study shows that C. salviniae is an effective biological control agent of S. molesta in semiarid areas.     

Soil Nitrogen Conditions Approach Preinvasion Levels following Restoration of Nitrogen-Fixing Black Locust (Robinia pseudoacacia) Stands in a Pine–Oak Ecosystem

Restoring native plant communities on sites formerly occupied by invasive nitrogen‐fixing species poses unique problems due to elevated soil nitrogen availability. Mitigation practices that reduce available nitrogen may ameliorate this problem. We evaluated the effects of tree removal followed by soil preparation or mulching on native plant growth and soil nitrogen transformations in a pine–oak system formerly occupied by exotic nitrogen‐fixing Black locust (Robinia pseudoacacia) trees. Greenhouse growth experiments with native grasses, Andropogon gerardii and Sorghastrum nutans, showed elevated relative growth rates in soils from Black locust compared with pine–oak stands. Field soil nutrient concentrations and rates of net nitrification and total net N‐mineralization were compared 2 and 4 years since Black locust removal and in control sites. Although soil nitrogen concentrations and total net N‐mineralization rates in the restored sites were reduced to levels that were similar to paired pine–oak stands after only 2 years, net nitrification rates remained 3–34 times higher in the restored sites. Other nutrient ion concentrations (Ca, Mg) and organic matter content were reduced, whereas phosphorus levels remained elevated in restored sites. Thus, 2–4 years following Black locust tree removal and soil horizon mixing achieved through site preparation, the concentrations of many soil nutrients returned to preinvasion levels. However, net nitrification rates remained elevated; cover cropping or carbon addition during restoration of sites invaded by nitrogen fixers could increase nitrogen immobilization and/or reduce nitrate availability, making sites more amenable to native plant establishment.     

Effects of calcium in the nitrogen-fixing symbiosis between actinorhizal <Emphasis Type="Italic">Discaria trinervis</Emphasis> (Rhamnaceae) and <Emphasis Type="Italic">Frankia</Emphasis>

We have analysed the growth and symbiotic performance of actinorhizal Discaria trinervis at various Ca supply regimes. We aimed at discriminating between specific, if any, effects on nodulation and general growth stimulation by Ca. The hypothesis that a high Ca supply would interfere with nodulation by Frankia was also tested. Results showed that plant growth increased with Ca supply. Nodulation was stimulated by moderate levels of Ca, but inhibited by Ca higher than 0.77 mM. Growth of nodules was less affected by Ca than shoot and root growth. Ca concentration of symbiotic plants increased with Ca supply, but nitrogen concentration was independent of Ca at concentrations which did not impair plant growth. All together, these results show that Ca has a positive effect on the establishment and functioning of the symbiosis between Discaria trinervis and Frankia. However, the positive influence of Ca was more likely due to a promotion of plant growth rather than a direct effect on nodule growth and nitrogen fixation itself. At high levels of Ca supply nodulation was impaired. Given the intercellular infection pathway in Discaria trinervis, we suggest that the increment of Ca availability would strengthen its root cell walls, thus decreasing Frankia penetration of the root.     

Seasonality in nutrient concentrations and stable isotope ratios of Halophila ovalis growing on the intertidal flat of SW Thailand

Halophila ovalis on the intertidal flat of SW Thailand maintains its presence under heavy grazing by dugongs; nevertheless, the site provides lower availability of nutrients in the water column to the seagrasses due to emersion during ebb tide. To examine the seasonality of the nutrient status of fast-growing H. ovalis in the intertidal flat, and to assess whether emersion at low tide affects the availability of nutrients and inorganic carbon, monthly changes in carbon (C), nitrogen (N), and phosphorus (P) concentrations of H. ovalis were determined. Carbon and nitrogen stable isotope ratios were also determined to examine the carbon and nitrogen sources. CNP concentrations of the leaves and petioles of H. ovalis at the intertidal flat were considerably lower than those of the same species previously reported from a subtidal environment throughout the observation period of 8 months. The δ¹³C of the above-ground parts of H. ovalis was lower than in previous reports, suggesting that CO₂ was sufficient for its growth. The CNP concentrations of H. ovalis on the intertidal flat increased after a decrease in grazing pressure by dugongs. It is suggested that H. ovalis sustained rapid growth on the intertidal flat by lowering its nutrient concentrations under heavy grazing by dugongs to maintain its presence. Received: March 2, 2001 / Accepted: June 6, 2001     

N2 fixation by Acacia species increases under elevated atmospheric CO2

In the present study the effect of elevated CO₂ on growth and nitrogen fixation of seven Australian Acacia species was investigated. Two species from semi‐arid environments in central Australia (Acacia aneura and A. tetragonophylla) and five species from temperate south‐eastern Australia (Acacia irrorata, A. mearnsii, A. dealbata, A. implexa and A. melanoxylon) were grown for up to 148 d in controlled greenhouse conditions at either ambient (350 µmol mol^?1) or elevated (700 µmol mol^?1) CO₂ concentrations. After establishment of nodules, the plants were completely dependent on symbiotic nitrogen fixation. Six out of seven species had greater relative growth rates and lower whole plant nitrogen concentrations under elevated versus normal CO₂. Enhanced growth resulted in an increase in the amount of nitrogen fixed symbiotically for five of the species. In general, this was the consequence of lower whole‐plant nitrogen concentrations, which equate to a larger plant and greater nodule mass for a given amount of nitrogen. Since the average amount of nitrogen fixed per unit nodule mass was unaltered by atmospheric CO₂, more nitrogen could be fixed for a given amount of plant nitrogen. For three of the species, elevated CO₂ increased the rate of nitrogen fixation per unit nodule mass and time, but this was completely offset by a reduction in nodule mass per unit plant mass.     

Nitrate levels affect the development of the black locust-Rhizobium symbiosis

Summary Experiments with black locust (Robinia pseudoacacia L.) seedlings grown under strictly controlled laboratory conditions indicated that the availability of nitrate has a marked impact on nitrogen fixation. When nitrate concentrations were very low, both nodulation and seedling growth were impaired, whereas nitrate concentrations high enough to promote plant growth strongly inhibited symbiotic nitrogen fixation. When nitrate was added to the growth medium after infection, nodulation and nitrogen fixation of the seedlings decreased. This effect was even more marked when nitrate was applied before infection with rhizobia. Higher nitrogen concentrations also reduced nodule number and nodule mass when applied simultaneously with the infecting bacteria. The contribution of symbiotic nitrogen fixation to black locust shoot mass by far exceeded its effects on shoot length and root mass. When nitrate availability was very low, specific nitrogen fixation (i. e. nitrogenase activity per nodule wet weight) was improved with increasing nitrogen supply, but rapidly decreased with higher nitrogen concentrations.     

Nodulation and growth response ofAllocasuarina andCasuarina species to phosphorus fertilization

To examine how soil phosphorus status affects nitrogen fixation by the Casuarinaceae —Frankia symbiosis,Casuarina equisetifolia and two species ofAllocasuarina (A. torulosa andA. littoralis) inoculated or fertilized with KNO₃ were grown in pots in an acid soil at 4 soil phosphate levels. InoculatedC. equisetifolia nodulated well by 12 weeks after planting and the numbers and weight of nodules increased markedly with phosphorus addition. Growth ofC. equisetifolia dependent on symbiotically fixed nitrogen was more sensitive to low levels of phosphorus (30 mg kg^–1 soil) than was growth of seedings supplied with combined nitrogen; at higher levels of phosphorus, the growth response curves were similar for both nitrogen fertilized and inoculated plants. The interaction between phosphorus and nitrogen treatments (inoculated and nitrogen fertilized) demonstrated that there was a greater requirement of phosphorus for symbiotic nitrogen fixation than for plant growth when soil phosphorus was low.WithAllocasuarina species, large plant to plant variation in nodulation occurred both within pots and between replicates. This result suggests genetic variation in nodulation withinAllocasuarina species. Nodulation ofAllocasuarina species did not start until 16 weeks after planting and no growth response due toFrankia inoculation was obtained at the time of harvest. Addition of nitrogen starter is suggested to boost plant growth before the establishment of the symbiosis. Growth ofAllocasuarina species fertilized with nitrogen responded to increasing levels of phosphorus up to 90 mg P/kg soil after which it declined by 69% forA. littoralis. The decrease in shoot weight ofA. littoralis, A. torulosa, C. equisetifolia andC. cunninghamiana at high phosphorus was confirmed in a sand culture experiment, and may be atributable to phosphorus toxicity.     

WATER RELATIONS AND GROWTH CHARACTERISTICS OF PROSOPIS GLANDULOSA VAR. TORREYANA IN A SIMULATED PHREATOPHYTIC ENVIRONMENT

Recent studies of Prosopis glandulosa have demonstrated a unique system of a deeply rooted species with significant water stress tolerance. Several growth and developmental characteristics have been correlated with water stress and nitrogen availability during field studies. Here we present a lab experiment in which a phreatophytic regime is simulated and the availability of nitrogen and water are varied. Increased ground water salinity caused lower plant water potentials and greater osmotic adjustment without significant increases in leaf Na⁺ concentrations. Leaf conductance was higher in the higher salinity treatments. Low water potential was also associated with reduced leaf size, reduced leaf area per plant and increased root to shoot ratio. Specific leaf weight and the transpiration ratio were unaffected by the low water potentials induced by increased salinity. Increasing nitrogen availability caused increased growth rates but did not influence water use efficiency. Net assimilation rates increased with increasing nitrogen availability but relative growth rates were more dependent on overall plant size than treatment conditions. The responses of P. glandulosa to the simulated phreatophytic environment were similar to those predicted by field measurements.     

Cost assessment of the production of pyrrolizidine alkaloids in ragwort (Senecio jacobaea L.)

Costs of pyrrolizidine alkaloid (Pa) production in vegetative ragwort (Senecio jacobaea) were examined under conditions in which plant growth was limited by light, nitrogen and phosphorus. Measurable costs of Pa production were demonstrated under light-limiting conditions. Plants with higher Pa concentrations grew more slowly than those with lower Pa concentration. Under nitrogen- and phosphorus-limited conditions no trade-off between Pa production and growth was observed.Publication of the Meijendel-comité, new series no. 116     

Effects of nitrogen deposition on the interaction between an aphid and its host plant

Abstract 1. Anthropogenic increases in nitrogen deposition are impacting terrestrial ecosystems worldwide. While some of the direct ecosystem‐level effects of nitrogen deposition are understood, the effects of nitrogen deposition on plant–insect interactions and on herbivore population dynamics have received less attention. 2. Nitrogen deposition will potentially influence both plant resource availability and herbivore population growth. If increases in herbivore population growth outstrip increases in resource availability, then increases in the strength of density dependence expressed within the herbivore population would be predicted. Alternatively, if plant resources respond more vigorously to nitrogen deposition than do herbivore populations, a decline in the strength of density dependence would be expected. No change in the strength of density dependence acting upon the herbivore population would suggest equivalent responses by herbivores and plants. 3. A density manipulation experiment was performed to examine the effect of nitrogen deposition on the interaction between a host plant, Asclepias tuberosa, and its herbivore, Aphis nerii. Aphid maximum per capita growth rate (R_max), carrying capacity (K), and the strength of density dependence were measured under three nitrogen deposition treatments. The effect of nitrogen deposition on the relationship among these three measures of insect population dynamics was explored. 4. Simulated nitrogen deposition increased aphid per capita population growth, plant foliar nitrogen concentrations, and plant biomass. Nitrogen deposition caused R_max and K to increase proportionally, leading to no overall change in the strength of density dependence. In this system, potential changes in the negative feedback processes operating on herbivore populations following nitrogen deposition appear to be buffered by concomitant changes in resource availability.     

How do Mycorrhizas Affect C and N Relationships in Flooded Aster tripolium Plants?

The mycorrhizal associations established between plants and fungi have multiple effects on plant growth, directly affecting stress tolerance. This work aimed to explore arbuscular mycorrhizal (AM) effects on carbon and nitrogen relationships of Aster tripolium L. and consequently on its flooding tolerance. Mycorrhizal and non-mycorrhizal juvenile plants were submitted to non-flooding and tidal flooding conditions for 56 d. Tidal flooding reduced biomass, but the presence of mycorrhiza had an ameliorating effect. The AM symbioses seem to have, like flooding, a stressful effect on A. tripolium at an early stage of plant development. However, once the plant was established, an improvement of growth performance of plants with mycorrhiza under flooding conditions was observed. The better tolerance of AM plants to flooding was mediated through an improvement of the osmotic adjustment of the plant tissues (higher concentrations of soluble sugars and proline) and through the increment of nitrogen acquisition in tidal-flooded plants.     

Elevated CO2 and plant nitrogen-use: is reduced tissue nitrogen concentration size-dependent?

Plants often respond to elevated atmospheric CO₂ levels with reduced tissue nitrogen concentrations relative to ambient CO₂-grown plants when comparisons are made at a common time. Another common response to enriched CO₂ atmospheres is an acceleration in plant growth rates. Because plant nitrogen concentrations are often highest in seedlings and subsequently decrease during growth, comparisons between ambient and elevated CO₂-grown plants made at a common time may not demonstrate CO₂-induced reductions in plant nitrogen concentration per se. Rather, this comparison may be highlighting differences in nitrogen concentration between bigger, more developed plants and smaller, less developed plants. In this study, we directly examined whether elevated CO₂ environments reduce plant nitrogen concentrations independent of changes in plant growth rates. We grew two annual plant species. Abutilon theophrasti (C₃ photosynthetic pathway) and Amaranthus retroflexus (C₄ photosynthetic pathway), from seed in glass-sided growth chambers with atmospheric CO₂ levels of 350 mol·mol^–1 or 700 mol·mol^–1 and with high or low fertilizer applications. Individual plants were harvested every 2 days starting 3 days after germination to determine plant biomass and nitrogen concentration. We found: 1. High CO₂-grown plants had reduced nitrogen concentrations and increased biomass relative to ambient CO₂-grown plants when compared at a common time; 2. Tissue nitrogen concentrations did not vary as a function of CO₂ level when plants were compared at a common size; and 3. The rate of biomass accumulation per rate of increase in plant nitrogen was unaffected by CO₂ availability, but was altered by nutrient availability. These results indicate that a CO₂-induced reduction in plant nitrogen concentration may not be due to physiological changes in plant nitrogen use efficiency, but is probably a size-dependent phenomenon resulting from accelerated plant growth.     

Nitrogen fixation byAlnus viridis (Chaix) DC.

Summary InAlnus viridis nodule growth relative to plant growth was inversely related to the quantity of nitrate added to nutrient solutions. Nodulated plants showed maximum growth when grown independently of supplied nitrogen and made better growth in its absence than unnodulated plants at any level of added nitrogen. Low levels of nitrate caused a depression of growth of nodulated plants, apparently by suppressing both nitrogen fixation and nodule growth. Nodules in nitrogen-free sand culture fixed atmospheric nitrogen at a rate of 6.6 mg/day/g nodule. Phosphorus deficiency was induced by low levels of phosphate and resulted in small plants with dark-green foliage. Root and nodule growth as a percentage of total plant growth and the percentage of total accumulated plant nitrogen below ground were greater at a root temperature of 11°C than 21°C. Thus at low root temperature processes other than nitrogen fixation were limiting to plant growth. Excised nodules were exposed to an N ₂ ¹⁵ -enriched atmosphere. A positive correlation between rate of nitrogen fixation and temperature was obtained, with optimum fixation occurring at about 20°C. It was shown that in spite of decreasing mean temperatures with increase in altitude, rate of nitrogen fixation by nodules of plants growing in the field increased with increase in altitude. This latter trend was deduced to be a reflection of the extent to which the field sites were nitrogen deficient in relation to climatically possible growth.