Effects of experimental nitrogen additions on plant diversity in an old-growth tropical forest

Response of plant biodiversity to increased availability of nitrogen (N) has been investigated in temperate and boreal forests, which are typically N-limited, but little is known in tropical forests. We examined the effects of artificial N additions on plant diversity (species richness, density and cover) of the understory layer in an N saturated old-growth tropical forest in southern China to test the following hypothesis: N additions decrease plant diversity in N saturated tropical forests primarily from N-mediated changes in soil properties. Experimental additions of N were administered at the following levels from July 2003 to July 2008: no addition (Control); 50 kg N ha−1 yr−1 (Low-N); 100 kg N ha−1 yr−1 (Medium-N), and 150 kg N ha−1 yr−1 (High-N). Results showed that no understory species exhibited positive growth response to any level of N addition during the study period. Although low-to-medium levels of N addition (≤100 kg N ha−1 yr−1) generally did not alter plant diversity through time, high levels of N addition significantly reduced species diversity. This decrease was most closely related to declines within tree seedling and fern functional groups, as well as to significant increases in soil acidity and Al mobility, and decreases in Ca availability and fine-root biomass. This mechanism for loss of biodiversity provides sharp contrast to competition-based mechanisms suggested in studies of understory communities in other forests. Our results suggest that high-N additions can decrease plant diversity in tropical forests, but that this response may vary with rate of N addition

Nitrogen input ¹⁵N signatures are reflected in plant ¹⁵N natural abundances in subtropical forests in China

Natural abundance of ¹⁵N (<i>δ</i>¹⁵N) in plants and soils can provide time-integrated information related to nitrogen (N) cycling within ecosystems, but it has not been well tested in warm and humid subtropical forests. In this study, we used ecosystem <i>δ</i>¹⁵N to assess effects of increased N deposition on N cycling in an old-growth broad-leaved forest and a secondary pine forest in a high-N-deposition area in southern China. We measured <i>δ</i>¹⁵N of inorganic N in input and output fluxes under ambient N deposition, and we measured N concentration (%N) and <i>δ</i>¹⁵N of major ecosystem compartments under ambient deposition and after decadal N addition at 50 kg N ha⁻¹yr⁻¹, which has a <i>δ</i>¹⁵N of −0.7 ‰. Our results showed that the total inorganic N in deposition was ¹⁵N-depleted (−10 ‰) mainly due to high input of strongly ¹⁵N-depleted NH₄⁺-N. Plant leaves in both forests were also ¹⁵N-depleted (−4 to −6 ‰). The broad-leaved forest had higher plant and soil %N and was more ¹⁵N-enriched in most ecosystem compartments relative to the pine forest. Nitrogen addition did not significantly affect %N in the broad-leaved forest, indicating that the ecosystem pools are already N-rich. However, %N was marginally increased in pine leaves and significantly increased in understory vegetation in the pine forest. Soil <i>δ</i>¹⁵N was not changed significantly by the N addition in either forest. However, the N addition significantly increased the <i>δ</i>¹⁵N of plants toward the ¹⁵N signature of the added N, indicating incorporation of added N into plants. Thus, plant <i>δ</i>¹⁵N was more sensitive to ecosystem N input manipulation than %N in these subtropical forests. We interpret the depleted <i>δ</i>¹⁵N of plants as an imprint from the high and ¹⁵N-depleted N deposition that may dominate the effects of fractionation that are observed in most warm and humid forests. Fractionation during the steps of N cycling could explain the difference between negative <i>δ</i>¹⁵N in plants and positive <i>δ</i>¹⁵N in soils, and the increase in soil <i>δ</i>¹⁵N with depths. Nevertheless, interpretation of ecosystem <i>δ</i>¹⁵N from high-N-deposition regions needs to include data on the deposition ¹⁵N signal

Generalist taxa shape fungal community structure in cropping ecosystems

Fungi regulate nutrient cycling, decomposition, symbiosis, and pathogenicity in cropland soils. However, the relative importance of generalist and specialist taxa in structuring soil fungal community remains largely unresolved. We hypothesized that generalist fungi, which are adaptable to various environmental conditions, could potentially dominate the community and become the basis for fungal coexisting networks in cropping systems. In this study, we identified the generalist and habitat specialist fungi in cropland soils across a 2,200 kms environmental gradient, including three bioclimatic regions (subtropical, warm temperate, and temperate). A few fungal taxa in our database were classified as generalist taxa (~1%). These generalists accounted for >35% of the relative abundance of all fungal populations, and most of them are Ascomycota and potentially pathotrophic. Compared to the specialist taxa (5–17% of all phylotypes in three regions), generalists had a higher degree of connectivity and were often identified as hub within the network. Structural equation modeling provided further evidence that after accounting for spatial and climatic/ edaphic factors, generalists had larger contributions to the fungal coexistence pattern than habitat specialists. Taken together, our study provided evidence that generalist taxa are crucial components for fungal community structure. The knowledge of generalists can provide important implication for understanding the ecological preference of fungal groups in cropland systems

Mineral N stock and nitrate accumulation in the 50 to 200 m profile on the Loess Plateau

Nitrogen (N) stored in deep profiles is important in assessing regional and/or global N stocks and nitrate leaching risk to groundwater. The Chinese Loess Plateau, which is characterized by significantly thick loess deposits, potentially stores immense stocks of mineral N, posing future threats to groundwater quality. In order to determine the vertical distributions of nitrate and ammonium content in the region, as well as to characterize the potential accumulation of nitrate in the deep loess profile, we study loess samples collected at five sites (Yangling, Changwu, Fuxian, An'sai and Shenmu) through a 50 to 200 m loess profile. The estimated storage of mineral N varied significantly among the five sites, ranging from 0.46 to 2.43 × 104 kg N ha−1. Ammonium exhibited fluctuations and dominated mineral N stocks within the whole profile at the sites, except for the upper 20–30 m at Yangling and Changwu. Measured nitrate content in the entire profile at Fuxian, An'sai and Shenmu is low, but significant accumulations were observed to 30–50 m depth at the other two sites. Analysis of δ15N and δ18O of nitrate indicates different causes for accumulated nitrate at these two sites. Mineralization and nitrification of manure and organic N respectively contribute nitrate to the 0–12 and 12–30 m profile at Changwu; while nitrification of NH4+ fertilizer, NO3− fertilizer and nitrification of organic N control the nitrate distribution in the 0–3, 3–7 and 7–10 m layer at Yangling, respectively. Furthermore, our analysis illustrates the low denitrification potential in the lower part of the vadose zone. The accumulated nitrate introduced by human activities is thus mainly distributed in the upper vadose zone (above 30 m), indicating, currently, a low nitrate leaching risk to groundwater due to a high storage capacity of the thick vadose zone in the region

A replicative recombinant HPV16 E7 expression virus upregulates CD36 in C33A cells

ObjectiveIn past decades, the role of high-risk HPV (HR-HPV) infection in cancer pathogenesis has been extensively studied. The viral E7 protein expressed in pre-malignant cells has been identified as an ideal target for immunological intervention. However, the cultivation of HPV in vitro remains a significant challenge, as well as the lack of methods for expressing the HPV E7 protein and generating replication-competent recombinant viral particles, which posed a major obstacle to further exploration of the function and carcinogenic mechanisms of the E7 oncoprotein. Therefore, it is imperative to investigate novel methodologies to construct replication-competent recombinant viral particles that express the HPV E7 protein to facilitate the study of its function.MethodsWe initiated the construction of recombinant viral particles by utilizing the ccdB-Kan forward/reverse screening system in conjunction with the Red/ExoCET recombinant system. We followed the infection of C33A cells with the obtained recombinant virus to enable the continuous expression of HPV16 E7. Afterwards, the total RNA was extracted and performed transcriptome sequencing using RNA-Seq technology to identify differentially expressed genes associated with HPV-induced oncogenicity.ResultsWe successfully established replicative recombinant viral particles expressing HPV16 E7 stably and continuously. The C33A cells were infected with recombinant viral particles to achieve overexpression of the E7 protein. Subsequently, RNA-Seq analysis was conducted to assess the changes in host cell gene expression. The results revealed an upregulation of the CD36 gene, which is associated with the HPV-induced oncogenic pathways, including PI3K-Akt and p53 signaling pathway. qRT-PCR analysis further identified that the upregulation of the CD36 gene due to the expression of HPV16 E7.ConclusionThe successful expression of HPV16 E7 in cells demonstrates that the replicated recombinant virus retains the replication and infection abilities of Ad4, while also upregulating the CD36 gene involved in the PI3K-Akt signaling and p53 pathways, thereby promoting cell proliferation. The outcome of this study provides a novel perspective and serves as a solid foundation for further exploration of HPV-related carcinogenesis and the development of replicative HPV recombinant vaccines capable of inducing protective immunity against HPV

First Assessment of NOx Sources at a Regional Background Site in North China Using Isotopic Analysis Linked with Modeling

important role in the formation of atmospheric particles. Thus, NOx emission reduction is critical for improving air quality, especially in severely air-polluted regions (e.g., North China). In this study, the source of NOx was investigated by the isotopic composition (delta N-15) of particulate nitrate (p-NO3-) at Beihuangcheng Island (BH), a regional background site in North China. It was found that the delta N-15-NO3- (n = 120) values varied between -1.7 parts per thousand and +24.0 parts per thousand and the delta O-18-NO3- values ranged from 49.4 parts per thousand to 103.9 parts per thousand. On the basis of the Bayesian mixing model, 27.78 +/- 8.89%, 36.53 +/- 6.66%, 22.01 +/- 6.92%, and 13.68 +/- 3.16% of annual NOx could be attributed to biomass burning, coal combustion, mobile sources, and biogenic soil emissions, respectively. Seasonally, the four sources were similar in spring and fall. Biogenic soil emissions were augmented in summer in association with the hot and rainy weather. Coal combustion increased significantly in winter with other sources showing an obvious decline. This study confirmed that isotope-modeling by delta N-15-NO3- is a promising tool for partitioning NOx sources and provides guidance to policymakers with regard to options for NOx reduction in North China

Revisiting the concentration observations and source apportionment of atmospheric ammonia

While China’s Air Pollution Prevention and Control Action Plan on particulate matter since 2013 has reduced sulfate significantly, aerosol ammonium nitrate remains high in East China. As the high nitrate abundances are strongly linked with ammonia, reducing ammonia emissions is becoming increasingly important to improve the air quality of China. Although satellite data provide evidence of substantial increases in atmospheric ammonia concentrations over major agricultural regions, long-term surface observation of ammonia concentrations are sparse. In addition, there is still no consensus on whether agricultural or non-agricultural emissions dominate the urban ammonia budget. Identifying the ammonia source by nitrogen isotope helps in designing a mitigation strategy for policymakers, but existing methods have not been well validated. Revisiting the concentration measurements and identifying source apportionment of atmospheric ammonia is thus an essential step towards reducing ammonia emissions

Spatial Variations of Soil N2 and N2O Emissions from a Temperate Forest: Quantified by the In Situ 15N Labeling Method

Emissions of dinitrogen (N2) and nitrous oxide (N2O) from soil are important components of the global nitrogen cycle. Soil N2O emissions from terrestrial ecosystems have been well studied. However, patterns and mechanisms of N2 emissions remain unclear due to the technical difficulty in measuring N2 production. In this study, an in situ 15N labeling method was employed to determine soil N2 and N2O emission rates from the lower, middle, and upper slopes, which correspond to different moisture conditions, in a temperate forest in Northeast China. We found that N2 emissions varied from 85 to 3442 &mu;g N m&minus;2 h&minus;1 across the slopes and were dominated by denitrification. The emissions of bulk N2O (22 to 258 &mu;g N m&minus;2 h&minus;1) and denitrification-derived N2O (14 to 246 &mu;g N m&minus;2 h&minus;1) were significantly lower than N2 emissions from their corresponding slope positions. Both N2 and N2O emissions significantly increased when soils become wetter. The ratios of N2O/(N2O + N2) were significantly higher at the upper and middle slopes (0.22 and 0.20, respectively) compared with those at the lower slope (0.08 &plusmn; 0.01). At the catchment scale, N2 accounted for 85% of the total gaseous N losses (N2O + N2). Our study shows that soil moisture drives the patterns of N2 and N2O emissions and field quantification of N2O/(N2O + N2) ratio should further consider the effect of slope position of forest ecosystems to estimate total soil gaseous N losses