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The recently discovered complete ammonia oxidation (comammox Nitrospira) containing clade A and clade B has further complemented our understanding of nitrification process. Nevertheless, understanding the community feature of comammox Nitrospira clades A and B and their relative contribution to nitrification in paddy rhizosphere are still in its infancy. In this study, we assessed the community diversity and structure of comammox Nitrospira clades A and B in paddy rhizosphere and bulk soils under thirty years of different fertilization strategies, i.e., non-fertilization control (CK), chemical fertilizers application (NPK), and NPK plus swine manure (NPKM), respectively. NPKM significantly increased the a-diversity (Chao1 and Shannon indices) of comammox Nitrospira clade A and altered the community structure (P < 0.05) but had little effect on clade B. A two-way analysis of variance (ANOVA) showed that the effect of long-term fertilization on soil comammox Nitrospira community and nitrification potential rate (PNR) was much greater than that of rhizosphere. Compared with NPK, soil PNR was greatly increased by 51.0% under the NPKM treatment in the rhizosphere (P < 0.05). Phylogenetic analysis showed that NPKM improved the relative abundances of sub-clade A.2.1 and sub-clade A.3.2 of the comammox clade A community, with an average increase of 212.2 and 210.4% in both rhizosphere and bulk soils relative to the NPK treatment. Soil organic matter, NH 4 -N, and pH were significant soil drivers of comammox Nitrospira clades A and B community. Furthermore, linear regression and structural equation modeling clearly showed that comammox Nitrospira clade A a-diversity were significantly associated with soil PNR (P < 0.05). Our results suggest (i) that comammox Nitrospira clade A are sensitive to the organic fertilization; and (ii) that comammox Nitrospira clade A contribute more to nitrification than clade B under the long-term organic fertilized paddy soil.+ -N, and pH were significant soil drivers of comammox Nitrospira clades A and B community. Furthermore, linear regression and structural equation modeling clearly showed that comammox Nitrospira clade A a-diversity were significantly associated with soil PNR (P < 0.05). Our results suggest (i) that comammox Nitrospira clade A are sensitive to the organic fertilization; and (ii) that comammox Nitrospira clade A contribute more to nitrification than clade B under the long-term organic fertilized paddy soil., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Aims: Continuous cropping is known to have profound effects on the soil microbial community in different planting systems. However, we lack an understanding of how different years of continuous cropping affects rhizosphere soil bacterial community co-occurrence pattern and assembly processes in the cut chrysanthemum (Chrysanthemum morifolium Ramat.) field., Methods and Results: We collected the soils from cut chrysanthemum rhizospheres with planting for 1 year (PY1) and continuous cropping for 6 years (CY6) and 12 years (CY12). Real-time quantitative PCR and flow cytometry (FCM) techniques were used to test the 16S rRNA gene copy number and bacterial cell count, respectively. The bacterial community structure was analysed by using high-throughput sequencing technology. The CY12 had a significantly decreased soil fertility index and rhizosphere bacterial living cell counts and gene copy numbers compared to CY6 and PY1 (P < 0.05). The rhizosphere bacterial community dissimilarity increased as the continuous cropping years increased. Three main ecological clusters (modules #1, #2, and #3) were observed in the bacterial co-occurrence network across all samples, and only the relative abundance of module #1 (enriched in the CY12) was significantly correlated with soil fertility (P < 0.05). Moreover, the rhizosphere bacterial community assembly was primarily governed by the deterministic process under 12 years of continuous cropping., Conclusions: Soil fertility decline correlates with ecological network modularization and the deterministic assembly process of the rhizosphere bacterial community of cut chrysanthemum during continuous cropping., (© The Author(s) 2023. Published by Oxford University Press on behalf of Applied Microbiology International.)

Different phosphorus (P) fertilizations significantly impact the transformation of the applied-P in soils. However, knowledge about how different P fertilization regimes influence the allocation of the amended-P in soil remains incomplete. Herein, we carried out a pot experiment to explore the fate of applied-P in calcareous soil using an oxygen isotope labeling technique ( 18 O-P 18 O 4 3- ). Treatments included check (CK), single, and repeated applications. The phosphorus mass balance result showed that more than 48.5% of the applied-P was held in labile and moderately labile fractions with the repeated treatment, while approximately 27.4% of the added-P was recovered in nonlabile forms in the single application treatment. The isotopic tracer ( 18 O-P 18 O 4 3- ) result demonstrated that the δ 18 O P values of NaHCO 3 -P and NaOH-P in the repeated P application were significantly higher than those in the single P application. Ultimately, better agronomic performances of the crops and higher PUE were achieved in the repeated treatment. Our findings highlighted that repeated P fertilization can improve P availability by reducing P fixation. These results pronounced that the enriched oxygen isotope technique can be considered an effective approach for tracing applied-P in soils.

Application of nitrification inhibitors (NIs) has been widely used to inhibit nitrification and reduce N 2 O emissions. However, the impacts of NI addition on soil carbon transformation and carbon-degrading microbial communities have not been well explored. Here, a microcosm experiment was carried out, and four treatments were designed: (i) unfertilized control, (ii) urea alone, (iii) urea plus cattle manure, and (iv) urea plus cattle manure with nitrapyrin. The influence of nitrapyrin on soil CO 2 emissions, carbon-degrading extracellular enzyme activities, and the abundance and diversity of the cbh I community was investigated. Compared to the treatment of urea plus cattle manure, nitrapyrin significantly decreased cumulative CO 2 emissions by 51.8%. Moreover, cbh I community gene copies and their α-diversities ( P < 0.05) were also significantly reduced by nitrapyrin application. A partial least squares path model showed that CO 2 emission was positively associated with cbh I community α-diversity but negatively associated with nitrapyrin addition. We conclude that the mitigation of soil CO 2 emissions by nitrapyrin can be ascribed to its effects on decreasing of cellulose-degrading gene community diversity. Our findings provide new insights into the side-effects of nitrapyrin on abating CO 2 emission.

Rain-induced soil CO 2 pulse, a rapid excitation in soil CO 2 flux after rain, is ubiquitously observed in terrestrial ecosystems, yet the underlying mechanisms in tropical forests are still not clear. We conducted a rain simulation experiment to quantify rain-induced changes in soil CO 2 flux and microbial community composition in a tropical forest. Soil CO 2 flux rapidly increased by ~83% after rains, accompanied by increases in both bacterial (~51%) and fungal (~58%) Phospholipid Fatty Acids (PLFA) biomass. However, soil CO 2 flux and microbial community in the plots without litters showed limited response to rains. Direct releases of CO 2 from litter layer only accounted for ~19% increases in soil CO 2 flux, suggesting that the leaching of dissolved organic carbon (DOC) from litter layer to the topsoil is the major cause of rain-induced soil CO 2 pulse. In addition, rain-induced changes in soil CO 2 flux and microbial PLFA biomass decreased with increasing rain sizes, but they were positively correlated with litter-leached DOC concentration rather than total DOC flux. Our findings reveal an important role of litter-leached DOC input in regulating rain-induced soil CO 2 pulses and microbial community composition, and may have significant implications for CO 2 losses from tropical forest soils under future rainfall changes.

With the continuing increase in anthropogenic activities, acid rain remains a serious environmental threat, especially in the fast developing areas such as southern China. To detect how prolonged deposition of acid rain would influence soil organic carbon accumulation in mature subtropical forests, we conducted a field experiment with simulated acid rain (SAR) treatments in a monsoon evergreen broadleaf forest at Dinghushan National Nature Reserve in southern China. Four levels of SAR treatments were set by irrigating plants with water of different pH values: CK (the control, local lake water, pH ≈ 4.5), T1 (water pH=4.0), T2 (water pH=3.5), and T3 (water pH=3.0). Results showed reduced pH measurements in the topsoil exposed to simulated acid rains due to soil acidification. Soil respiration, soil microbial biomass and litter decomposition rates were significantly decreased by the SAR treatments. As a result, T3 treatment significantly increased the total organic carbon by 24.5% in the topsoil compared to the control. Furthermore, surface soil became more stable as more recalcitrant organic matter was generated under the SAR treatments. Our results suggest that prolonged acid rain exposure may have the potential to facilitate soil organic carbon accumulation in the subtropical forest in southern China., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Phytoliths represent one of the few available altitudinal vegetation proxies for mountain ecosystems. This study analyzed 41 topsoil phytolith samples collected from five altitudinal zones in the southern Himalaya as far as, and beyond, the timberline, from tropical forest (up to 1,000 m a.s.l.) to subtropical forest (1,000-2,000 m a.s.l.), to temperate forest (2,000-3,000 m a.s.l.), to subalpine forest (3,000-4,100 m a.s.l.) and finally to alpine scrub (4,100-5,200 m a.s.l.). The statistical results show a good correlation between phytolith assemblages and these five altitudinal vegetation zones: the five phytolith assemblages identified effectively differentiated these five altitudinal vegetation zones. In particular, coniferous phytoliths accurately indicated the timberline. Additionally, we tested the phytolith index Ic (a proxy for estimating the percentage of Pooideae vis-à-vis the total grass content) as a quantifier of phytolith variety versus altitude. Ic increased along altitude, as expected. An investigation of phytoliths provided an initial basis for the analysis of the composition of gramineous vegetation. Furthermore, redundancy analysis and discriminant analysis also suggested a significant correlation between phytolith assemblages and altitude. Our research therefore provides an up-to-date analogue for the reconstruction of changes to palaeovegetation and palaeoaltitude in mountainous areas.

Acid rain is an environmental problem of increasing concern in China. In this study, a laboratory leaching column experiment with acid forest soil was set up to investigate the responses of soil and soil solution chemistry to simulated acid rain (SAR). Five pH levels of SAR were set: 2.5, 3.0, 3.5, 4.0, and 4.5 (as a control, CK). The results showed that soil acidification would occur when the pH of SAR was ≤3.5. The concentrations of NO₃(-)and Ca(2+) in the soil increased significantly when the pH of SAR fell 3.5. The concentration of SO₄(2-) in the soil increased significantly when the pH of SAR was <4.0. The effects of SAR on soil solution chemistry became increasingly apparent as the experiment proceeded (except for Na(+) and dissolved organic carbon (DOC)). The net exports of NO₃(-), SO₄(2-), Mg(2+), and Ca(2+) increased about 42-86% under pH 2.5 treatment as compared to CK. The Ca(2+) was sensitive to SAR, and the soil could release Ca(2+) through mineral weathering to mitigate soil acidification. The concentration of exchangeable Al(3+) in the soil increased with increasing the acidity of SAR. The releases of soluble Al and Fe were SAR pH dependent, and their net exports under pH 2.5 treatment were 19.6 and 5.5 times, respectively, higher than that under CK. The net export of DOC was reduced by 12-29% under SAR treatments as compared to CK. Our results indicate the chemical constituents in the soil are more sensitive to SAR than those in the soil solution, and the effects of SAR on soil solution chemistry depend not only on the intensity of SAR but also on the duration of SAR addition. The soil and soil solution chemistry in this region may not be affected by current precipitation (pH≈4.5) in short term, but the soil and soil leachate chemistry may change dramatically if the pH of precipitation were below 3.5 and 3.0, respectively.

Background: Natural forest succession often affects soil physical and chemical properties. Selected physical and chemical soil properties were studied in an old-growth forest across a forest successional series in Dinghushan Nature Reserve, Southern China., Methodology/principal Findings: The aim was to assess the effects of forest succession change on soil properties. Soil samples (0-20 cm depth) were collected from three forest types at different succession stages, namely pine (Pinus massoniana) forest (PMF), mixed pine and broadleaf forest (PBMF) and monsoon evergreen broadleaf forest (MEBF), representing early, middle and advanced successional stages respectively. The soil samples were analyzed for soil water storage (SWS), soil organic matter (SOM), soil microbial biomass carbon (SMBC), pH, NH4(+)-N, available potassium (K), available phosphorus (P) and microelements (available copper (Cu), available zinc (Zn), available iron (Fe) and available boron (B)) between 1999 and 2009. The results showed that SWS, SOM, SMBC, Cu, Zn, Fe and B concentrations were higher in the advanced successional stage (MEBF stage). Conversely, P and pH were lower in the MEBF but higher in the PMF (early successional stage). pH, NH4(+)-N, P and K declined while SOM, Zn, Cu, Fe and B increased with increasing forest age. Soil pH was lower than 4.5 in the three forest types, indicating that the surface soil was acidic, a stable trend in Dinghushan., Conclusion/significance: These findings demonstrated significant impacts of natural succession in an old-growth forest on the surface soil nutrient properties and organic matter. Changes in soil properties along the forest succession gradient may be a useful index for evaluating the successional stages of the subtropical forests. We caution that our inferences are drawn from a pseudo-replicated chronosequence, as true replicates were difficult to find. Further studies are needed to draw rigorous conclusions regarding on nutrient dynamics in different successional stages of forest.

The response of soil respiration to acid rain in forests, especially in forests of different maturity, is poorly understood in southern China despite the fact that acid rain has become a serious environmental threat in this region in recent years. Here, we investigated this issue in three subtropical forests of different maturity [i.e. a young pine forest (PF), a transitional mixed conifer and broadleaf forest (MF) and an old-growth broadleaved forest (BF)] in southern China. Soil respiration was measured over two years under four simulated acid rain (SAR) treatments (CK, the local lake water, pH 4.5; T1, water pH 4.0; T2, water pH 3.5; and T3, water pH 3.0). Results indicated that SAR did not significantly affect soil respiration in the PF, whereas it significantly reduced soil respiration in the MF and the BF. The depressed effects on both forests occurred mostly in the warm-wet seasons and were correlated with a decrease in soil microbial activity and in fine root biomass caused by soil acidification under SAR. The sensitivity of the response of soil respiration to SAR showed an increasing trend with the progressive maturity of the three forests, which may result from their differences in acid buffering ability in soil and in litter layer. These results indicated that the depressed effect of acid rain on soil respiration in southern China may be more pronounced in the future in light of the projected change in forest maturity. However, due to the nature of this field study with chronosequence design and the related pseudoreplication for forest types, this inference should be read with caution. Further studies are needed to draw rigorous conclusions regarding the response differences among forests of different maturity using replicated forest types.

Background: The aim of this study was to determine response patterns and mechanisms of soil respiration to precipitation increases in subtropical regions., Methodology/principal Findings: Field plots in three typical forests [i.e. pine forest (PF), broadleaf forest (BF), and pine and broadleaf mixed forest (MF)] in subtropical China were exposed under either Double Precipitation (DP) treatment or Ambient Precipitation (AP). Soil respiration, soil temperature, soil moisture, soil microbial biomass and fine root biomass were measured over three years. We tested whether precipitation treatments influenced the relationship of soil respiration rate (R) with soil temperature (T) and soil moisture (M) using R = (a+cM)exp(bT), where a is a parameter related to basal soil respiration; b and c are parameters related to the soil temperature and moisture sensitivities of soil respiration, respectively. We found that the DP treatment only slightly increased mean annual soil respiration in the PF (15.4%) and did not significantly change soil respiration in the MF and the BF. In the BF, the increase in soil respiration was related to the enhancements of both soil fine root biomass and microbial biomass. The DP treatment did not change model parameters, but increased soil moisture, resulting in a slight increase in soil respiration. In the MF and the BF, the DP treatment decreased soil temperature sensitivity b but increased basal soil respiration a, resulting in no significant change in soil respiration., Conclusion/significance: Our results indicate that precipitation increasing in subtropical regions in China may have limited effects on soil respiration.

This study was designed to examine the responses of soil CO(2) efflux to precipitation pulses of varying intensities using precipitation simulations in two subtropical forests [i.e., mixed and broadleaf forests (MF and BF)] in southern China. The artificial precipitation event was achieved by spraying a known amount of water evenly in a plot (50 × 50 cm(2)) over a 30 min period, with intensities ranging from 10, 20, 50 and 100 mm within the 30 min. The various intensities were simulated in both dry season (in December 2007) and wet (in May 2008) season. We characterized the dynamic patterns of soil CO(2) efflux rate and environmental factors over the 5 h experimental period. Results showed that both soil moisture and soil CO(2) efflux rate increased to peak values for most of the simulated precipitation treatments, and gradually returned to the pre-irrigation levels after irrigation in two forests. The maximum peak of soil CO(2) efflux rate occurred at the 10 mm precipitation event in the dry season in BF and was about 3.5 times that of the pre-irrigation value. The change in cumulative soil CO(2) efflux following precipitation pulses ranged from -0.68 to 1.72 g CO(2) m(-2) over 5 h compared to the pre-irrigation levels and was generally larger in the dry season than in the wet season. The positive responses of soil CO(2) efflux to precipitation pulses declined with the increases in precipitation intensity, and surprisingly turned to negative when precipitation intensity reached 50 and 100 mm in the wet season. These findings indicated that soil CO(2) efflux could be changed via pulse-like fluxes in subtropical forests in southern China as fewer but extreme precipitation events occur in the future.

The total flux and concentration of total organic carbon (TOC) in hydrological processes in coniferous and broad-leaved mixed forest of Dinghushan were measured from July 2002 to July 2003. The results showed that the TOC input by precipitation was 41.80 kg x hm(-2) x yr(-1), while its output by surface runoff and groundwater (soil solution at 50 cm depth) was 17.54 and 1.80 kg x hm(-2) x yr(-1), respectively. The difference between input and output was 22.46 kg x hm(-2) x yr(-1), indicating that the ecosystem TOC was in positive balance. The monthly variation of TOC flux in hydrological processes was very similar to that in precipitation. The mean TOC concentration in precipitation was 3.64 mg x L(-1), while that in throughfall and stemflow increased 6.10 and 7.39 times after rain passed through the tree canopies and barks. The mean TOC concentration in surface runoff and in soil solution at 25 and 50 cm depths was 12.72, 7.905 and 3.06 mg x L(-1), respectively. The monthly TOC concentration in throughfall and stemflow had a similar changing tendency, showing an increase at the beginning of growth season (March), a decrease after September, and a little increase in December. The TOC concentration in runoff was much higher during high precipitation months. No obvious monthly variation was observed in soil solution TOC concentration (25 and 50 cm below the surface). Stemflow TOC concentration differed greatly between different tree species. The TOC concentration in precipitation, throughfall, and soil solution (25 and 50 cm depths) decreased with increasing precipitation, and no significant relationship existed between the TOC concentrations in stemflow, surface runoff and precipitation. The TOC concentrations in the hydrological processes fluctuated with precipitation intensity, except for that in stemflow and soil solutions.