Effect of Agricultural Management on Nematode Communities in a Mediterranean Agroecosystem

The effects of agricultural management on the soil nematode community were investigated in a field study at depths of 0 to 10 cm and 10 to 20 cm during a peanut (Arachis hypogaea) growing season in Israel. Nineteen nematode families and 23 genera were observed. Rhabditidae, Cephalobus, Eucephalobus, Aphelenchus, Aphelenchoides, Tetylenchus, Tylenchus, Dorylaimus, and Discolaimus were the dominant family and genera. Ecological measures of soil nematode community structure, diversity, and maturity indices were assessed and compared between the managed (by fertilization, irrigation, and pesticide application) and unmanaged fields. The total number of nematodes at a 10-cm depth during peanut-sowing, mid-season, and harvest periods was higher in the treated (managed) plot than in the control (unmanaged) plot. Bacterivores and fungivores were the most abundant trophic groups in both plots and both depths. The relative abundance of each group averaged 60.8 to 67.3% and 11.5 to 19.6% of the nematode community, respectively. Plant parasites and omnivores-predators at the 0 to 10-cm depth were much less abundant than any other two groups in our experimental plots. During the growing season, except the harvest period, populations of plant parasites and omnivores-predators at the 10 to 20-cm depth were lower in the treated plot than in the control plot. Maturity index (MI), plant-parasite index (PPI), and ratio of fungivores and bacterivores to plant parasites (WI) were found to be more sensitive indicators than other ecological indices for assessing the response of nematode communities to agricultural management in an Israeli agroecosystem.     

Nitrogen Fertilization Has a Stronger Effect on Soil Nitrogen-Fixing Bacterial Communities than Elevated Atmospheric CO2

Biological nitrogen fixation is the primary supply of N to most ecosystems, yet there is considerable uncertainty about how N-fixing bacteria will respond to global change factors such as increasing atmospheric CO₂ and N deposition. Using the nifH gene as a molecular marker, we studied how the community structure of N-fixing soil bacteria from temperate pine, aspen, and sweet gum stands and a brackish tidal marsh responded to multiyear elevated CO₂ conditions. We also examined how N availability, specifically, N fertilization, interacted with elevated CO₂ to affect these communities in the temperate pine forest. Based on data from Sanger sequencing and quantitative PCR, the soil nifH composition in the three forest systems was dominated by species in the Geobacteraceae and, to a lesser extent, Alphaproteobacteria. The N-fixing-bacterial-community structure was subtly altered after 10 or more years of elevated atmospheric CO₂, and the observed shifts differed in each biome. In the pine forest, N fertilization had a stronger effect on nifH community structure than elevated CO₂ and suppressed the diversity and abundance of N-fixing bacteria under elevated atmospheric CO₂ conditions. These results indicate that N-fixing bacteria have complex, interacting responses that will be important for understanding ecosystem productivity in a changing climate.     

Morphology and tissue quality of seedling root systems of Pinus taeda and Pinus ponderosa as affected by varying CO2, temperature,and nitrogen

Rising atmospheric carbon dioxide, nitrogen deposition and warmer temperatures may alter the quantity and quality of plant-derived organic matter available to soil biota, potentially altering rates of belowground herbivory and decomposition. Our objective was to simulate future growth conditions for an early successional (loblolly) and late successional (ponderosa) species of pine to determine if the physical and chemical properties of the root systems would change. Seedlings were grown for 160 days in greenhouses at the Duke University Phytotron at 35 or 70 Pa CO₂ partial pressure, ambient or ambient + 5 °C temperature, and 1 or 5 mMNH₄O₃. Roots from harvested seedlings were analyzed for changes in surface area, specific root length, mass, total nonstructural carbohydrates (TNC), and concentrations of macro-nutrients. Surface area increased in both species under elevated CO₂, due primarily to increases in root length, and this response was greatest (+138%) in loblolly pine at high temperature. Specific root length decreased in loblolly pine at elevated CO₂ but increases in mass more than compensated for this, resulting in net increases in total length. TNC was unaffected and nutrient concentrations decreased only slightly at elevated CO₂, possibly from anatomical changes to the root tissues. We conclude that future growth conditions will enhance soil exploration by some species of pine, but root carbohydrate levels and nutrient concentrations will not be greatly affected, leaving rates of root herbivory and decomposition unaltered.     

Effects of elevated carbon dioxide and nitrogen fertilization on nitrate reductase activity in sweetgum and loblolly pine trees in two temperate forests

Nitrogen (N) availability is a major factor limiting plant production in many terrestrial ecosystems and is a key regulator of plant response to elevated CO₂. Plant N status is a function of both soil N availability and plant N uptake and assimilation capacity. As a rate-limiting step in nitrate assimilation, the reduction of nitrate is an important component of plant physiological response to elevated CO₂ and terrestrial carbon sequestration. We examine the effects of elevated CO₂ and N availability on the activity of nitrate reductase, the enzyme catalyzing the reduction of nitrate to nitrite, in two temperate forests—a closed canopy sweetgum (Liquidambar styraciflua) plantation in Tennessee (Oak Ridge National Laboratory (ORNL)) and a loblolly pine (Pinus taeda) stand in North Carolina (Duke). Both CO₂ and N enrichment had species specific impacts on nitrate reductase activity (NaR). Elevated CO₂ and N fertilization decreased foliar NaR in P. taeda, but there were no treatment effects on L. styraciflua NaR at ORNL or Duke. NaR in 1-year P. taeda needles was significantly greater than in 0-year old needles across treatments. P. taeda NaR was negatively correlated with bio-available molybdenum concentrations in soils, suggesting that CO₂ and N-mediated changes in soil nutrient status may be altering soil-plant N-dynamics. The variation in response among species may reflect different strategies for acquiring N and suggests that elevated CO₂ may alter plant N dynamics through changes in NaR.     

Temperature- and moisture-induced changes in the structure of the nematode fauna of a semiarid grassland –- patterns and mechanisms

A field experiment was established to examine the effects of temperature and moisture modifications on the nematode fauna of a semiarid grassland. Several combinations of drying, wetting, warming and cooling were applied to plots and compared with untreated control plots. The experiment was performed from July to October 1996. A significant shift was observed in the structure of the nematode fauna between late summer and early autumn. This shift was manifested in the disappearance of four rare genera; Ecumenicus, Eucephalobus, Paraphelenchus and Pungentus. A significant decrease was found in the density of Acrobeles, Aphelenchoides, Ditylenchus and Prismatolaimus; in addition there was a significant increase in the density of Cephalobus, Helicotylenchus, Paratylenchus and Tylenchorhynchus. Community structural change was represented by an initial decrease in nematode generic richness of 30–50%, and in a statistically significant decrease of nematode diversity in the control and all treated plots. Thereafter, emergence of a new community was demonstrated. Data show that temperature manipulation was the main factor to influence nematode diversity, Maturity Index, and Plant Parasite Index. However, nematode population density was influenced predominantly by the soil moisture content. Coenological analysis of soil nematode fauna appears to be a useful tool for the biological monitoring of the effects of global change on semiarid grasslands.     

Six years of in situ CO2 enrichment evoke changes in soil structure and soil biota of nutrient-poor grassland

Nutrient‐poor grassland on a silty clay loam overlying calcareous debris was exposed to elevated CO₂ for six growing seasons. The CO₂ exchange and productivity were persistently increased throughout the experiment, suggesting increases in soil C inputs. At the same time, elevated CO₂ lead to increased soil moisture due to reduced evapotransporation. Measurements related to soil microflora did not indicate increased soil C fluxes under elevated CO₂. Microbial biomass, soil basal respiration, and the metabolic quotient for CO₂ (qCO₂) were not altered significantly. PLFA analysis indicated no significant shift in the ratio of fungi to bacteria. 0.5 m KCl extractable organic C and N, indicators of changed DOC and DON concentrations, also remained unaltered. Microbial grazer populations (protozoa, bacterivorous and fungivorous nematodes, acari and collembola) and root feeding nematodes were not affected by elevated CO₂. However, total nematode numbers averaged slightly lower under elevated CO₂ (?16%, ns) and nematode mass was significantly reduced (?43%, P = 0.06). This reduction reflected a reduction in large‐diameter nematodes classified as omnivorous and predacious. Elevated CO₂ resulted in a shift towards smaller aggregate sizes at both micro‐ and macro‐aggregate scales; this was caused by higher soil moisture under elevated CO₂. Reduced aggregate sizes result in reduced pore neck diameters. Locomotion of large‐diameter nematodes depends on the presence of large enough pores; the reduction in aggregate sizes under elevated CO₂ may therefore account for the decrease in large nematodes. These animals are relatively high up the soil food web; this decline could therefore trigger top‐down effects on the soil food web. The CO₂ enrichment also affected the nitrogen cycle. The N stocks in living plants and surface litter increased at elevated CO₂, but N in soil organic matter and microbes remained unaltered. Nitrogen mineralization increased markedly, but microbial N did not differ between CO₂ treatments, indicating that net N immobilization rates were unaltered. In summary, this study did not provide evidence that soils and soil microbial communities are affected by increased soil C inputs under elevated CO₂. On the contrary, available data (¹³C tracer data, minirhizotron observations, root ingrowth cores) suggests that soil C inputs did not increase substantially. However, we provide first evidence that elevated CO₂ can reduce soil aggregation at the scale from µ m to mm scale, and that this can affect soil microfaunal populations.     

Canopy N and P dynamics of a southeastern US pine forest under elevated CO2

Forest production is strongly nutrient limited throughout the southeastern US. If nutrient limitations constrain plant acquisition of essential resources under elevated CO₂, reductions in the mass or nutrient content of forest canopies could constrain C assimilation from the atmosphere. We tested this idea by quantifying canopy biomass, foliar concentrations of N and P, and the total quantity of N and P in a loblolly pine (Pinus taeda) canopy subject to 4 years of free-air CO₂ enrichment. We also used N:P ratios to detect N versus P limitation to primary production under elevated CO₂. Canopy biomass was significantly higher under elevated CO₂ during the first 4 years of this experiment. Elevated CO₂ significantly reduced the concentration of N in loblolly pine foliage (5% relative to ambient CO₂) but not P. Despite the slight reduction foliage N concentrations, there were significant increases in canopy N and P contents under elevated CO₂. Foliar N:P ratios were not altered by elevated CO₂ and were within a range suggesting forest production is N limited not P limited. Despite the clear limitation of NPP by N under ambient and elevated CO₂ at this site, there is no evidence that the mass of N or P in the canopy is declining through the first 4 years of CO₂ fumigation. As a consequence, whole-canopy C assimilation is strongly stimulated by elevated CO₂ making this forest a larger net C sink under elevated CO₂ than under ambient CO₂. We discuss the potential for future decreases in canopy nutrient content as a result of limited changes in the size of the plant-available pools of N under elevated CO₂.     

Atmospheric CO2 enrichment increases growth and photosynthesis of Pinus taeda: a 4 year experiment in the field

Forest trees are major components of the terrestrial biome and their response to rising atmospheric CO₂ plays a prominent role in the global carbon cycle. In this study, loblolly pine seedlings were planted in the field in recently disturbed soil of high fertility, and CO₂ partial pressures were maintained at ambient CO₂ (Amb) and elevated CO₂ (Amb + 30 Pa) for 4 years. The objective of the study was to measure seasonal and long-term responses in growth and photosynthesis of loblolly pine exposed to elevated CO₂ under ambient field conditions of precipitation, light, temperature and nutrient availability. Loblolly pine trees grown in elevated CO₂ produced 90% more biomass after four growing seasons than did trees grown in ambient CO₂. This large increase in final biomass was primarily due to a 217% increase in leaf area in the first growing season which resulted in much higher relative growth rates for trees grown in elevated CO₂. Although there was not a sustained effect of elevated CO₂ on relative growth rate after the first growing season, absolute production of biomass continued to increase each year in trees grown in elevated CO₂ as a consequence of the compound interest effect of increased leaf area on the production of more new leaf area and more biomass. Allometric analyses of biomass allocation patterns demonstrated size-dependent shifts in allocation, but no direct effects of elevated CO₂ on partitioning of biomass. Leaf photosynthetic rates were always higher in trees grown in elevated CO₂, but these differences were greater in the summer (60–130% increase) than in the winter (14–44% increase), reflecting strong seasonal effects of temperature on photosynthesis. Our results suggest that seasonal variation in the relative photosynthetic response to elevated CO₂ will occur in natural ecosystems, but total non-structural carbohydrate (TNC) levels in leaves indicate that this variation may not always be related to sink activity. Despite indications of canopy-level adjustments in carbon assimilation, enhanced levels of leaf photosynthesis coupled with increased total leaf area indicate that net carbon assimilation for the whole tree was greater for trees grown under elevated CO₂ compared with ambient CO₂. If the large growth enhancement observed in loblolly pine were maintained after canopy closure, then these trees could be a large sink for fossil carbon emitted to the atmosphere and produce a negative feedback on atmospheric CO₂.     

A Taxonomic Review of the Genera of the Aphelenchoidea (Fuchs, 1937) Thorne, 1949 (Nematoda: Tylenchida)

This generic level taxonomic review of the nematode superfamily Aphelenchoidea is based upon a study of specimens from 24 of the 28 described genera. The diagnoses of these genera are presented, and some are emended with new information. One new genus, Huntaphelenchoides, and five new combinations are proposed. The families Paraphelenchidae and Anomyctidae are placed in synonymy with Aphelenchidae and Aphelenchoididae, respectively. The genera Asteroaphelenchoides and Pseudaphelencboides are placed in synonymy under the genus Aphelenchoides. Rare male and/or female specimens of Peraphelenchus, Anomyctus, Laimaphelenchus, Aphelenchus, Cryptaphelenchoides, Megadorus, Tylaphelenchus, and Entaphelenchus, are redescribed and illustrated. Four plates, containing 106 original drawings of the males, females, stylets, and spicules of representatives of 22 aphelenchoid genera, are presented.     

Photosynthetic capacity of loblolly pine (Pinus taeda L.) trees during the first year of carbon dioxide enrichment in a forest ecosystem

Our objective was to assess the photosynthetic responses of loblolly pine trees (Pinus taeda L.) during the first full growth season (1997) at the Brookhaven National Lab/Duke University Free Air CO₂ Enrichment (FACE) experiment. Gas exchange, fluorescence characteristics, and leaf biochemistry of ambient CO₂ (control) needles and ambient + 20 Pa CO₂ (elevated) needles were examined five times during the year. The enhancement of photosynthesis by elevated CO₂ in mature loblolly pine trees varied across the season and was influenced by abiotic and biotic factors. Photosynthetic enhancement by elevated CO₂ was strongly correlated with leaf temperature. The magnitude of photosynthetic enhancement was zero in March but was as great as 52% later in the season. In March, reduced sink demand and lower temperatures resulted in lower net photosynthesis, lower carboxylation rates and higher excess energy dissipation from the elevated CO₂ needles than from control needles. The greatest photosynthetic enhancement by CO₂ enrichment was observed in July during a period of high temperature and low precipitation, and in September during recovery from this period of low precipitation. In July, loblolly pine trees in the control rings exhibited lower net photosynthetic rates, lower maximum rates of photosynthesis at saturating CO₂ and light, lower values of carboxylation and electron transport rates (modelled from A–C_i curves), lower total Rubisco activity, and lower photochemical quenching of fluorescence in comparison to other measurement periods. During this period of low precipitation trees in the elevated CO₂ rings exhibited reduced net photosynthesis and photochemical quenching of fluorescence, but there was little effect on light- and CO₂-saturated rates of photosynthesis, modelled rates of carboxylation or electron transport, or Rubisco activity. These first-year data will be used to compare with similar measurements from subsequent years of the FACE experiment in order to determine whether photosynthetic acclimation to CO₂ occurs in these canopy loblolly pine trees growing in a forest ecosystem.     

Photosynthetic responses to CO2 enrichment of four hardwood species in a forest understory

We compared the CO₂- and light-dependence of photosynthesis of four tree species (Acer rubrum, Carya glabra, Cercis canadensis, Liquidambar styraciflua) growing in the understory of a loblolly pine plantation under ambient or ambient plus 200 μl l^–1 CO₂. Naturally-established saplings were fumigated with a free-air CO₂ enrichment system. Light-saturated photosynthetic rates were 159–190% greater for Ce. canadensis saplings grown and measured under elevated CO₂. This species had the greatest CO₂ stimulation of photosynthesis. Photosynthetic rates were only 59% greater for A. rubrum saplings under CO₂ enrichment and Ca. glabra and L. styraciflua had intermediate responses. Elevated CO₂ stimulated light-saturated photosynthesis more than the apparent quantum yield. The maximum rate of carboxylation of ribulose-1,5-bisphosphate carboxylase, estimated from gas-exchange measurements, was not consistently affected by growth in elevated CO₂. However, the maximum electron transport rate estimated from gas- exchange measurements and from chlorophyll fluorescence, when averaged across species and dates, was approximately 10% higher for saplings in elevated CO₂. The proportionately greater stimulation of light-saturated photosynthesis than the apparent quantum yield and elevated rates of maximum electron transport suggests that saplings growing under elevated CO₂ make more efficient use of sunflecks. The stimulation of light-saturated photosynthesis by CO₂ did not appear to correlate with shade-tolerance ranking of the individual species. However, the species with the greatest enhancement of photosynthesis, Ce. canadensis and L. styraciflua, also invested the greatest proportion of soluble protein in Rubisco. Environmental and endogenous factors affecting N partitioning may partially explain interspecific variation in the photosynthetic response to elevated CO₂. Received: 16 February 1999 / Accepted: 30 August 1999     

First-year growth response of trees in an intact forest exposed to elevated CO2

Although elevated atmospheric CO₂ has been shown to increase growth of tree seedlings and saplings, the response of intact forest ecosystems and established trees is unclear. We report results from the first large-scale experimental system designed to study the effects of elevated CO₂ on an intact forest with the full complement of species interactions and environmental stresses. During the first year of exposure to ^ 1.5 Ë ambient CO₂, canopy loblolly pine (Pinus taeda, L.) trees increased basal area growth rate by 24% but understorey trees of loblolly pine, sweetgum (Liquidambar styraciflua L.), and red maple (Acer rubrum L.) did not respond. Winged elm (Ulmus alata Michx.) had a marginally significant increase in growth rate (P = 0.069). These data suggest that this ecosystem has the capacity to respond immediately to a step increase in atmospheric CO₂; however, as exposure time increases, nutrient limitations may reduce this initial growth stimulation.     

A field study of the effects of elevated CO2 on carbon assimilation, stomatal conductance and leaf and branch growth of Pinus taeda trees

A study was conducted in 21-year-old loblolly pine (Pinus taeda L.) trees growing in plantation in north central Georgia, USA. The experiment used branch chambers to impose treatments of ambient, ambient +165 and ambient + 330 μmol mol^?1 CO₂. After one growing season there was no indication of acclimation to elevated CO₂. In August and September, carbon assimilation, measured by two different methods, was twice as high at ambient +330 μmol mol^?1 CO₂ than at ambient. Dark respiration was suppressed by 6% at ambient +165 and by 14% at ambient + 330 μmol mol^?1 CO₂. This suppression was immediate, and not an effect of exposure to elevated CO₂ during growth, since respiration was reduced by the same amount in all treatments when measured at a high CO₂ concentration. Elevated CO₂ increased the growth of foliage and woody tissue. It also increased instantaneous transpiration efficiency, but it had no effect on stomatal conductance. Since the soil at the study site had low to moderate fertility, these results suggest that the growth potential of forests on many sites may be enhanced by global increases in atmospheric CO₂, concentration.     

Ocean Acidification Affects the Phyto-Zoo Plankton Trophic Transfer Efficiency

The critical role played by copepods in ocean ecology and biogeochemistry warrants an understanding of how these animals may respond to ocean acidification (OA). Whilst an appreciation of the potential direct effects of OA, due to elevated pCO₂, on copepods is improving, little is known about the indirect impacts acting via bottom-up (food quality) effects. We assessed, for the first time, the chronic effects of direct and/or indirect exposures to elevated pCO₂ on the behaviour, vital rates, chemical and biochemical stoichiometry of the calanoid copepod Acartia tonsa. Bottom-up effects of elevated pCO₂ caused species-specific biochemical changes to the phytoplanktonic feed, which adversely affected copepod population structure and decreased recruitment by 30%. The direct impact of elevated pCO₂ caused gender-specific respiratory responses in A.tonsa adults, stimulating an enhanced respiration rate in males (> 2-fold), and a suppressed respiratory response in females when coupled with indirect elevated pCO₂ exposures. Under the combined indirect+direct exposure, carbon trophic transfer efficiency from phytoplankton-to-zooplankton declined to < 50% of control populations, with a commensurate decrease in recruitment. For the first time an explicit role was demonstrated for biochemical stoichiometry in shaping copepod trophic dynamics. The altered biochemical composition of the CO₂-exposed prey affected the biochemical stoichiometry of the copepods, which could have ramifications for production of higher tropic levels, notably fisheries. Our work indicates that the control of phytoplankton and the support of higher trophic levels involving copepods have clear potential to be adversely affected under future OA scenarios.     

Soil–Nitrogen Cycling in a Pine Forest Exposed to 5 Years of Elevated Carbon Dioxide

Empirical and modeling studies have shown that the magnitude and duration of the primary production response to elevated carbon dioxide (CO₂) can be constrained by limiting supplies of soil nitrogen (N). We have studied the response of a southern US pine forest to elevated CO₂ for 5 years (1997–2001). Net primary production has increased significantly under elevated CO₂. We hypothesized that the increase in carbon (C) fluxes to the microbial community under elevated CO₂ would increase the rate of N immobilization over mineralization. We tested this hypothesis by quantifying the pool sizes and fluxes of inorganic and organic N in the forest floor and top 30 cm of mineral soil during the first 5 years of CO₂ fumigation. We observed no statistically significant change in the gross or net rate of inorganic N mineralization and immobilization in any soil horizon under elevated CO₂. Similarly, elevated CO₂ had no statistically significant effect on the concentration or flux of organic N, including amino acids. Microbial biomass N was not significantly different between CO₂ treatments. Thus, we reject our hypothesis that elevated CO₂ increases the rate of N immobilization. The quantity and chemistry of the litter inputs to the forest floor and mineral soil horizons can explain the limited range of microbially mediated soil–N cycling responses observed in this ecosystem. Nevertheless a comparative analysis of ecosystem development at this site and other loblolly pine forests suggests that rapid stand development and C sequestration under elevated CO₂ may be possible only in the early stages of stand development, prior to the onset of acute N limitation.     

Soil nematode abundances drive agroecosystem multifunctionality under short-term elevated CO2 and O3

The response of soil biotas to climate change has the potential to regulate multiple ecosystem functions. However, it is still challenging to accurately predict how multiple climate change factors will affect multiple ecosystem functions. Here, we assessed the short-term responses of agroecosystem multifunctionality to a factorial combination of elevated CO₂ (+200 ppm) and O₃ (+40 ppb) and identified the key soil biotas (i.e., bacteria, fungi, protists, and nematodes) concerning the changes in the multiple ecosystem functions for two rice varieties (Japonica, Nanjing 5055 vs. Wuyujing 3). We provided strong evidence that combined treatment rather than individual treatments of short-term elevated CO₂ and O₃ significantly increased the agroecosystem multifunctionality index by 32.3% in the Wuyujing 3 variety, but not in the Nanjing 5055 variety. Soil biotas exhibited an important role in regulating multifunctionality under short-term elevated CO₂ and O₃, with soil nematode abundances better explaining the changes in ecosystem multifunctionality than soil biota diversity. Furthermore, the higher trophic groups of nematodes, omnivores-predators served as the principal predictor of agroecosystem multifunctionality. These results provide unprecedented new evidence that short-term elevated CO₂ and O₃ can potentially affect agroecosystem multifunctionality through soil nematode abundances, especially omnivores-predators. Our study demonstrates that high trophic groups were specifically beneficial for regulating multiple ecosystem functions and highlights the importance of soil nematode communities for the maintenance of agroecosystem functions and health under climate change in the future.     

Molecular Characterization and Development of Real-Time PCR Assay for Pine-Wood Nematode Bursaphelenchus xylophilus (Nematoda: Parasitaphelenchidae)

Bursaphelenchus xylophilus, the pine-wood nematode (PWN), is the causal agent of pine wilt disease, one of the most damaging emerging pest problems to forests around the world. It is native to North America where it causes relatively minor damage to native conifers but is labeled an EPPO-A-2 pest and a quarantine nematode for many countries outside of the United States because of its potential for destruction to their native conifers. Exports of wood logs and commodities involving softwood packaging materials now require a lab test for the presence/absence of this regulated nematode species. We characterized the DNA sequences on the ribosomal DNA small subunit, large subunit D2/D3, internal transcribed spacer (ITS) and mitochondrial DNA cytochrome oxidase subunit one on the aphelenchid species and described the development of a real-time-PCR method for rapid and accurate identification of PWN targeting the ITS-1. A total of 97 nematode populations were used to evaluate the specificity and sensitivity of this assay, including 45 populations of B. xylophilus and 36 populations of 21 other species of Bursaphelenchus which belong to the abietinus, cocophilus, eggersi, fungivorus, hofmanni, kevini, leoni, sexdentati, and xylophilus groups and one unassigned group from a total of 13 groups in the genus Bursaphelenchus; 15 populations of Aphelenchoides besseyi, A. fragariae, Aphelenchoides species and Aphelenchus avenae; and one population of mixed nematode species from a soil sample. This assay proved to be specific to B. xylophilus only and was sensitive to a single nematode specimen regardless of the life stages present. This approach provides rapid species identification necessary to comply with the zero-tolerance export regulations.     

Elevated CO2 and aboveground–belowground herbivory by the clover root weevil

Predicted increases in atmospheric carbon dioxide (CO₂) concentrations are expected to increase primary productivity in many terrestrial ecosystems, which could lead to plants becoming N limited. Studies suggest that legumes may partially overcome this by increasing biological nitrogen fixation. However, these studies have not yet considered how these changes may be affected by the altered dynamics of insect herbivores feeding on the plant. This study investigated how elevated CO₂ (700 μl l^?1) affected the clover root weevil (Sitona lepidus), a significant pest of white clover (Trifolium repens). Adults feed on leaves aboveground where they lay eggs; soil-dwelling larvae initially feed on root nodules that house N₂-fixing bacteria. Foliar C:N ratios rose by 9% at elevated CO₂, but the biggest responses were observed belowground, with increases in root mass (85% greater) and nodule abundance (220% more abundant). Root C:N ratios increased significantly from 10.95 to 11.60 under elevated CO₂, which increased even further to 13.13 when nodules were attacked by larval S. lepidus. Adult S. lepidus consumed significantly more leaf tissue at elevated CO₂ (0.47 cm² day^?1) compared with ambient CO₂ (0.35 cm² day^?1), suggesting compensatory feeding, but laid 23% fewer eggs at elevated CO₂. Even though fewer eggs were laid at elevated CO₂, 38% more larvae were recovered suggesting that larval survival was much better under elevated CO₂. Increased larval abundance and performance at elevated CO₂ were positively correlated with the number of nodules available. In conclusion, reduced foliar quality at elevated CO₂ was generally disadvantageous for adult S. lepidus living aboveground, but extremely beneficial for S. lepidus larvae living belowground, due to the enhanced nodulation. Climate change may, therefore, enhance biological nitrogen fixation by T. repens, but potential benefits (e.g. provision of N without chemical fertilizers) may be undermined by larger populations of S. lepidus larvae belowground.     

Construction cost of loblolly and ponderosa pine leaves grown with varying carbon and nitrogen availability

We grew loblolly and ponderosa pine seedlings in a factorial experiment with two CO₂ partial pressures (35 and 70 Pa), and two nitrogen treatments (1.0 and 3.5 mol m^?3 NH₄⁺), for one growing season to examine the effects of carbon and nitrogen availability on leaf construction cost. Growth in elevated CO₂ reduced leaf nitrogen concentrations by 17 to 40%, and increased C:N by 22 to 68%. Elevated N availability increased leaf N concentrations and decreased C:N. Non-structural carbohydrates increased in high-CO₂-grown loblolly seedlings, except in fascicles from low N, and in ponderosa primary and fascicle leaves grown in high N. In loblolly, increases in starch were nearly 2-fold greater than the increases in soluble sugars. In ponderosa, only the soluble sugars were affected by CO₂. Leaf construction cost (g glucose g^?1 dm) varied by 9.3% across all treatments. All of the variation in loblolly leaf construction cost could be explained by changes in non-structural carbohydrates. A model of the response of construction cost to changes in the mass of different biochemical fractions suggests that the remainder of the variation in ponderosa, not explained by non-structural carbohydrates, is probably attributable to changes in lignin, phenolic or protein concentrations.     

Temporal dynamics and spatial variability in the enhancement of canopy leaf area under elevated atmospheric CO2

Increased canopy leaf area (L) may lead to higher forest productivity and alter processes such as species dynamics and ecosystem mass and energy fluxes. Few CO₂ enrichment studies have been conducted in closed canopy forests and none have shown a sustained enhancement of L. We reconstructed 8 years (1996–2003) of L at Duke's Free Air CO₂ Enrichment experiment to determine the effects of elevated atmospheric CO₂ concentration ([CO₂]) on L before and after canopy closure in a pine forest with a hardwood component, focusing on interactions with temporal variation in water availability and spatial variation in nitrogen (N) supply. The dynamics of L were reconstructed using data on leaf litterfall mass and specific leaf area for hardwoods, and needle litterfall mass and specific leaf area combined with needle elongation rates, and fascicle and shoot counts for pines. The dynamics of pine L production and senescence were unaffected by elevated [CO₂], although L senescence for hardwoods was slowed. Elevated [CO₂] enhanced pine L and the total canopy L (combined pine and hardwood species; P<0.050); on average, enhancement following canopy closure was ～16% and 14% respectively. However, variation in pine L and its response to elevated [CO₂] was not random. Each year pine L under ambient and elevated [CO₂] was spatially correlated to the variability in site nitrogen availability (e.g. r²=0.94 and 0.87 in 2001, when L was highest before declining due to droughts and storms), with the [CO₂]‐induced enhancement increasing with N (P=0.061). Incorporating data on N beyond the range of native fertility, achieved through N fertilization, indicated that pine L had reached the site maximum under elevated [CO₂] where native N was highest. Thus closed canopy pine forests may be able to increase leaf area under elevated [CO₂] in moderate fertility sites, but are unable to respond to [CO₂] in both infertile sites (insufficient resources) and sites having high levels of fertility (maximum utilization of resources). The total canopy L, representing the combined L of pine and hardwood species, was constant across the N gradient under both ambient and elevated [CO₂], generating a constant enhancement of canopy L. Thus, in mixed species stands, L of canopy hardwoods which developed on lower fertility sites (～3 g N inputs m^?2 yr^?1) may be sufficiently enhanced under elevated [CO₂] to compensate for the lack of response in pine L, and generate an appreciable response of total canopy L (～14%).