Plectus of the Prairie: A Case Study of Taxonomic Resolution from a Nematode Biodiversity Survey.

Taxonomic resolution is a critical component of biodiversity assessments. In this case study, we examined a single taxon within a larger study of nematode diversity to evaluate the taxonomic resolution of different diversity assessment methods. The selected taxon was the microbial-feeding genus Plectus, a group considered to include multiple cosmopolitan species. The methods included a morphological evaluation by light microscopy, Sanger sequencing of PCR amplicons of COI and 18S gene regions, and 18S metabarcoding sequencing. The study sites were 15 remnant tallgrass prairie plots in eastern Nebraska. In the morphological analysis, we observed two basic morphotypes, a short-tailed form with a small amphid and a long-tailed form with a large amphid. Sanger sequencing of COI sorted Plectus diversity into six distinct clades. The largest two of these six clades keyed to P. parietinus and P. rhizophilus based on morphology. BLAST analysis with COI revealed no close matches in GenBank. Sanger sequencing of the 18S region did not differentiate the six clades. These results illustrate that the method of diversity assessment strongly influences estimates of biodiversity. An additional 95 Plectus specimens, from outside the remnant sites, added taxonomic breadth to the COI phylogenetic tree. There were no geographically widespread COI haplotypes and no evidence of cosmopolitan Plectus species.

Fire legacies in eastern ponderosa pine forests.

Disturbance legacies structure communities and ecological memory, but due to increasing changes in disturbance regimes, it is becoming more difficult to characterize disturbance legacies or determine how long they persist. We sought to quantify the characteristics and persistence of material legacies (e.g., biotic residuals of disturbance) that arise from variation in fire severity in an eastern ponderosa pine forest in North America. We compared forest stand structure and understory woody plant and bird community composition and species richness across unburned, low-, moderate-, and high-severity burn patches in a 27-year-old mixed-severity wildfire that had received minimal post-fire management. We identified distinct tree densities (high: 14.3 ± 7.4 trees per ha, moderate: 22.3 ± 12.6, low: 135.3 ± 57.1, unburned: 907.9 ± 246.2) and coarse woody debris cover (high: 8.5 ± 1.6% cover per 30 m transect, moderate: 4.3 ± 0.7, low: 2.3 ± 0.6, unburned: 1.0 ± 0.4) among burn severities. Understory woody plant communities differed between high-severity patches, moderate- and low-severity patches, and unburned patches (all p < 0.05). Bird communities differed between high- and moderate-severity patches, low-severity patches, and unburned patches (all p < 0.05). Bird species richness varied across burn severities: low-severity patches had the highest (5.29 ± 1.44) and high-severity patches had the lowest (2.87 ± 0.72). Understory woody plant richness was highest in unburned (5.93 ± 1.10) and high-severity (5.07 ± 1.17) patches, and it was lower in moderate- (3.43 ± 1.17) and low-severity (3.43 ± 1.06) patches. We show material fire legacies persisted decades after the mixed-severity wildfire in eastern ponderosa forest, fostering distinct structures, communities, and species in burned versus unburned patches and across fire severities. At a patch scale, eastern and western ponderosa system responses to mixed-severity fires were consistent.

Assessment of Selected Antibiotic Resistances in Ungrazed Native Nebraska Prairie Soils.

The inherent spatial heterogeneity and complexity of antibiotic-resistant bacteria and antibiotic resistance (AR) genes in manure-affected soils makes it difficult to sort out resistance that can be attributed to human antibiotic use from resistance that occurs naturally in the soil. This study characterizes native Nebraska prairie soils that have not been affected by human or food-animal waste products to provide data on background levels of resistance in southeastern Nebraskan soils. Soil samples were collected from 20 sites enumerated on tetracycline and cefotaxime media; screened for tetracycline-, sulfonamide-, ß-lactamase-, and macrolide-resistance genes; and characterized for soil physical and chemical parameters. All prairies contained tetracycline- and cefotaxime-resistant bacteria, and 48% of isolates collected were resistant to two or more antibiotics. Most (98%) of the soil samples and all 20 prairies had at least one tetracycline gene. Most frequently detected were (D), (A) (O), (L), and (B). Sulfonamide genes, which are considered a marker of human or animal activity, were detected in 91% of the samples, despite the lack of human inputs at these sites. No correlations were found between either phenotypic or genotypic resistance and soil physical or chemical parameters. Heterogeneity was observed in AR within and between prairies. Therefore, multiple samples are necessary to overcome heterogeneity and to accurately assess AR. Conclusions regarding AR depend on the gene target measured. To determine the impacts of food-animal antibiotic use on resistance, it is essential that background and/or baseline levels be considered, and where appropriate subtracted out, when evaluating AR in agroecosystems.

Grassland productivity limited by multiple nutrients.

Terrestrial ecosystem productivity is widely accepted to be nutrient limited(1). Although nitrogen (N) is deemed a key determinant of aboveground net primary production (ANPP)(2,3), the prevalence of co-limitation by N and phosphorus (P) is increasingly recognized(4-8). However, the extent to which terrestrial productivity is co-limited by nutrients other than N and P has remained unclear. Here, we report results from a standardized factorial nutrient addition experiment, in which we added N, P and potassium (K) combined with a selection of micronutrients (K+µ), alone or in concert, to 42 grassland sites spanning five continents, and monitored ANPP. Nutrient availability limited productivity at 31 of the 42 grassland sites. And pairwise combinations of N, P, and K+µ co-limited ANPP at 29 of the sites. Nitrogen limitation peaked in cool, high latitude sites. Our findings highlight the importance of less studied nutrients, such as K and micronutrients, for grassland productivity, and point to significant variations in the type and degree of nutrient limitation. We suggest that multiple-nutrient constraints must be considered when assessing the ecosystem-scale consequences of nutrient enrichment.

Seasonal changes in depth of water uptake for encroaching trees Juniperus virginiana and Pinus ponderosa and two dominant C4 grasses in a semiarid grassland.

We used the natural abundance of stable isotopic ratios of hydrogen and oxygen in soil (0.05-3 m depth), plant xylem and precipitation to determine the seasonal changes in sources of soil water uptake by two native encroaching woody species (Pinus ponderosa P. & C. Lawson, Juniperus virginiana L.), and two C(4) grasses (Schizachyrium scoparium (Michx.) Nash, Panicum virgatum L.), in the semiarid Sandhills grasslands of Nebraska. Grass species extracted most of their water from the upper soil profile (0.05-0.5 m). Soil water uptake from below 0.5 m depth increased under drought, but appeared to be minimal in relation to the total water use of these species. The grasses senesced in late August in response to drought conditions. In contrast to grasses, P. ponderosa and J. virginiana trees exhibited significant plasticity in sources of water uptake. In winter, tree species extracted a large fraction of their soil water from below 0.9 m depth. In spring when shallow soil water was available, tree species used water from the upper soil profile (0.05-0.5 m) and relied little on water from below 0.5 m depth. During the growing season (May-August) significant differences between the patterns of tree species water uptake emerged. Pinus ponderosa acquired a large fraction of its water from the 0.05-0.5 and 0.5-0.9 m soil profiles. Compared with P. ponderosa, J. virginiana acquired water from the 0.05-0.5 m profile during the early growing season but the amount extracted from this profile progressively declined between May and August and was mirrored by a progressive increase in the fraction taken up from 0.5-0.9 m depth, showing plasticity in tracking the general increase in soil water content within the 0.5-0.9 m profile, and being less responsive to growing season precipitation events. In September, soil water content declined to its minimum, and both tree species shifted soil water uptake to below 0.9 m. Tree transpiration rates (E) and water potentials (Psi) indicated that deep water sources did not maintain E which sharply declined in September, but played an important role in the recovery of tree Psi. Differences in sources of water uptake among these species and their ecological implications on tree-grass dynamics and soil water in semiarid environments are discussed.

Species and functional group diversity independently influence biomass accumulation and its response to CO2 and N.

The characteristics of plant assemblages influence ecosystem processes such as biomass accumulation and modulate terrestrial responses to global change factors such as elevated atmospheric CO(2) and N deposition, but covariation between species richness (S) and functional group richness (F) among assemblages obscures the specific role of each in these ecosystem responses. In a 4-year study of grassland species grown under ambient and elevated CO(2) and N in Minnesota, we experimentally varied plant S and F to assess their independent effects. We show here that at all CO(2) and N levels, biomass increased with S, even with F constant at 1 or 4 groups. Likewise, with S at 4, biomass increased as F varied continuously from 1 to 4. The S and F effects were not dependent upon specific species or functional groups or combinations and resulted from complementarity. Biomass increases in response to CO(2) and N, moreover, varied with time but were generally larger with increasing S (with F constant) and with increasing F (with S constant). These results indicate that S and F independently influence biomass accumulation and its response to elevated CO(2) and N.

Nitrogen mineralization dynamics in grass monocultures.

Although Wedin and Tilman (1990) observed large differences in in situ N mineralization among monocultures of five grass species, the mechanisms responsible were unclear. In this study, we found that the species did not change total soil C or N, and soil C: N ratio (range 12.9-14.1) was only slightly, but significantly, changed after four years. Nor did the species significantly affect the total amount of N mineralized (per g soil N) in year-long aerobic laboratory incubations. However, short-term N mineralization rates in the incubations (day 1-day 17) differed significantly among species and were significantly correlated with annual in situ mineralization. When pool sizes and turnover rates of potentially mineralizable N (No) were estimated, the best model treated No as two pools: a labile pool, which differed among species in size (Nl, range 2-3% of total N) and rate constant (h, range 0.04-0.26 wk-1), and a larger recalcitrant pool with a constant mineralization rate across species. The rate constant of the labile pool (h) was highly correlated with annual in situ N mineralization (+0.96). Therefore, plant species need only change the dynamics of a small fraction of soil organic matter, in this case estimated to be less than 3%, to have large effects on overall system N dynamics.

Species effects on nitrogen cycling: a test with perennial grasses.

To test for differing effects of plant species on nitrogen dynamics, we planted monocultures of five perennial grasses (Agropyron repens, Agrostis scabra, Poa pratensis, Schizachyrium scoparium, and Andropogon gerardi) on a series of soils ranging from sand to black soil. In situ net N mineralization was measured in the monocultures for three years. By the third year, initially identical soils under different species had diverged up to 10-fold in annual net mineralization. This divergence corresponded to differences in the tissue N concentrations, belowground lignin concentrations, and belowground biomasses of the species. These results demonstrate the potential for strong feedbacks between the species composition of vegetation and N cycling. If individual plant species can affect N mineralization and N availability, then competition for N may lead to positive or negative feedbacks between the processes controlling species composition and ecosystem processes such as N and C cycling. These feedbacks create the potential for alternative stable states for the vegetation-soil system given the same initial abiotic conditions.