Long-term nitrogen and phosphorus addition have stronger negative effects on microbial residual carbon in subsoils than topsoils in subtropical forests.

Highly weathered lowland (sub)tropical forests are widely recognized as nitrogen (N)-rich and phosphorus (P)-poor, and the input of N and P affects soil carbon (C) cycling and storage in these ecosystems. Microbial residual C (MRC) plays a crucial role in regulating soil organic C (SOC) stability in forest soils. However, the effects of long-term N and P addition on soil MRC across different soil layers remain unclear. This study conducted a 12-year N and P addition experiment in two typical subtropical plantation forests dominated by Acacia auriculiformis and Eucalyptus urophylla trees, respectively. We measured plant C input (fine root biomass, fine root C, and litter C), microbial community structure, enzyme activity (C/N/P-cycling enzymes), mineral properties, and MRC. Our results showed that continuous P addition reduced MRC in the subsoil (20-40 cm) of both plantations (A. auriculiformis: 28.44% and E. urophylla: 28.29%), whereas no significant changes occurred in the topsoil (0-20 cm). N addition decreased MRC in the subsoil of E. urophylla (25.44%), but had no significant effects on A. auriculiformis. Combined N and P addition reduced MRC (34.63%) in the subsoil of A. auriculiformis but not in that of E. urophylla. The factors regulating MRC varied across soil layers. In the topsoil (0-10 cm), plant C input (the relative contributions to the total variance was 20%, hereafter) and mineral protection (47.2%) were dominant factors. In the soil layer of 10-20 cm, both microbial characteristics (41.3%) and mineral protection (32.3%) had substantial effects, whereas the deeper layer (20-40 cm) was predominantly regulated by microbial characteristics (37.9%) and mineral protection (18.8%). Understanding differential drivers of MRC across soil depth, particularly in deeper soil layers, is crucial for accurately predicting the stability and storage of SOC and its responses to chronic N enrichment and/or increased P limitation in (sub)tropical forests.

Negative effects of long-term phosphorus additions on understory plants in a primary tropical forest.

Human activities have disturbed global phosphorus (P) cycling by introducing substantial amounts of P to natural ecosystems. Although natural P gradients and fertilization studies have found that plant community traits are closely related to P availability, it remains unclear how increased P supply affects plant growth and diversity in P-deficient tropical forests. We used a decadal P-addition experiment (2007-2017) to study the effects of increased P input on plant growth and diversity in understory layer in tropical forests. We monitored the dynamics of seedling growth, survival rate, and diversity of understory plants throughout the fertilization period under control and P addition at 15 g P m-2 yr-1. To identify the drivers of responses, P concentration, photosynthesis rate and nonstructural carbon were analyzed. Results showed that long-term P addition significantly increased P concentrations both in soil pools and plant tissues. However, P addition did not increase the light-saturated photosynthesis rate or growth rate of the understory plants. Furthermore, P addition significantly decreased the survival rate of seedlings and reduced the species richness and density of understory plants. The negative effects of P addition may be attributed to an increased carbon cost due to the tissue maintenance of plants with higher P concentrations. These findings indicate that increased P supply alone is not necessary to benefit the growth of plants in ecosystems with low P availability, and P inputs can inhibit understory plants and may alter community composition. Therefore, we appeal to a need for caution when inputting P to tropical forests ecosystems.

Effects of human disturbance activities and environmental change factors on terrestrial nitrogen fixation.

Biological nitrogen (N) fixation plays an important role in terrestrial N cycling and represents a key driver of terrestrial net primary productivity (NPP). Despite the importance of N fixation in terrestrial ecosystems, our knowledge regarding the controls on terrestrial N fixation remains poor. Here, we conducted a meta-analysis (based on 852 observations from 158 studies) of N fixation across three types of ecosystems with different status of disturbance (no management, restoration [previously disturbed], and disturbance [currently disturbed]) and in response to multiple environmental change factors (warming, elevated carbon dioxide [CO2 ], increased precipitation, increased drought, increased N deposition, and their combinations). We explored the mechanisms underlying the changes in N fixation by examining the variations in soil physicochemical properties (bulk density, texture, moisture, and pH), plant and microbial characteristics (dominant plant species numbers, plant coverage, and soil microbial biomass), and soil resources (total carbon, total N, total phosphorus (P), inorganic N, and inorganic P). Human disturbance inhibited non-symbiotic N fixation but not symbiotic N fixation. Terrestrial N fixation was stimulated by warming (+152.7%), elevated CO2 (+19.6%), and increased precipitation (+73.1%) but inhibited by increased drought (-30.4%), N deposition (-31.0%), and combinations of available multiple environmental change factors (-14.5%), the extents of which varied among biomes and ecosystem compartments. Human disturbance reduced the N fixation responses to environmental change factors, which was associated with the changes in soil physicochemical properties (2%-56%, p < .001) and the declines in plant and microbial characteristics (3%-49%, p ≤ .003) and soil resources (6%-48%, p ≤ .03). Overall, our findings reveal for the first time the effects of multiple environmental change factors on terrestrial N fixation and indicate the role of human disturbance activities in inhibiting N fixation, which can improve our understanding, modeling, and prediction of terrestrial N budgets, NPP, and ecosystem feedbacks under global change scenarios.

Substrate stoichiometry determines nitrogen fixation throughout succession in southern Chinese forests.

The traditional view holds that biological nitrogen (N) fixation often peaks in early- or mid-successional ecosystems and declines throughout succession based on the hypothesis that soil N richness and/or phosphorus (P) depletion become disadvantageous to N fixers. This view, however, fails to support the observation that N fixers can remain active in many old-growth forests despite the presence of N-rich and/or P-limiting soils. Here, we found unexpected increases in N fixation rates in the soil, forest floor, and moss throughout three successional forests and along six age-gradient forests in southern China. We further found that the variation in N fixation was controlled by substrate carbon(C) : N and C : (N : P) stoichiometry rather than by substrate N or P. Our findings highlight the utility of ecological stoichiometry in illuminating the mechanisms that couple forest succession and N cycling.

Effects of urbanization on plant phosphorus availability in broadleaf and needleleaf subtropical forests.

Urbanization, the migration of populations from rural to urban areas, has been causing great stress on natural environments, leading to air pollution and nitrogen (N) deposition, negatively affecting forest health. Although there is evidence that urbanization has changed forest N cycling, little is known about whether urbanization also changes the availability of phosphorus (P), which is important for plant growth and forest productivity. To address this question, we carried out a survey in the Pearl River Delta region, the world's largest urban area in southern China, using two types of representative forests, the evergreen broadleaf forests (BFs) and pine plantations (PPs). The leaf N:P ratios in the two forest types were high (20-50) with a significant increasing pattern along the rural-to-urban gradient. The ratios of leaf P:K and P:Na declined along the rural-to-urban gradient, whereas leaf P content did not change in BF but decreased in PP along the rural-to-urban gradient, suggesting that leaf P became limiting along urbanization. The abundance of actinomycetes and gram-negative bacteria decreased along the rural-to-urban gradient, indicating the negative effects of urbanization on soil microorganisms. Principal component analysis indicated that divergent key factors respond to the urbanization and affect plant P limitation in the two forest types. In BF, broadleaf trees showed a greater response to N deposition from urbanization indicating direct leaf N uptake from N deposition is a key factor for plant P limitation. Alternatively, in PP, our findings suggest soil acidification is an important factor accelerating plant P limitation. Our study revealed that urbanization intensifies plant P limitation in subtropical forests, and the effects vary depending on forest types. Our findings provide empirical information to support the management of forest ecosystems and evaluation of urbanization effects on forest health.

Effects of nitrogen and phosphorus additions on soil microbial biomass and community structure in two reforested tropical forests.

Elevated nitrogen (N) deposition may aggravate phosphorus (P) deficiency in forests in the warm humid regions of China. To our knowledge, the interactive effects of long-term N deposition and P availability on soil microorganisms in tropical replanted forests remain unclear. We conducted an N and P manipulation experiment with four treatments: control, N addition (15 g N m(-2)·yr(-1)), P addition (15 g P m(-2)·yr(-1)), and N and P addition (15 + 15 g N and P m(-2)·yr(-1), respectively) in disturbed (planted pine forest with recent harvests of understory vegetation and litter) and rehabilitated (planted with pine, but mixed with broadleaf returning by natural succession) forests in southern China. Nitrogen addition did not significantly affect soil microbial biomass, but significantly decreased the abundance of gram-negative bacteria PLFAs in both forest types. Microbial biomass increased significantly after P addition in the disturbed forest but not in the rehabilitated forest. No interactions between N and P additions on soil microorganisms were observed in either forest type. Our results suggest that microbial growth in replanted forests of southern China may be limited by P rather than by N, and this P limitation may be greater in disturbed forests.

Nutrient limitation in three lowland tropical forests in southern China receiving high nitrogen deposition: insights from fine root responses to nutrient additions.

Elevated nitrogen (N) deposition to tropical forests may accelerate ecosystem phosphorus (P) limitation. This study examined responses of fine root biomass, nutrient concentrations, and acid phosphatase activity (APA) of bulk soil to five years of N and P additions in one old-growth and two younger lowland tropical forests in southern China. The old-growth forest had higher N capital than the two younger forests from long-term N accumulation. From February 2007 to July 2012, four experimental treatments were established at the following levels: Control, N-addition (150 kg N ha(-1) yr(-1)), P-addition (150 kg P ha(-1) yr(-1)) and N+P-addition (150 kg N ha(-1) yr(-1) plus 150 kg P ha(-1) yr(-1)). We hypothesized that fine root growth in the N-rich old-growth forest would be limited by P availability, and in the two younger forests would primarily respond to N additions due to large plant N demand. Results showed that five years of N addition significantly decreased live fine root biomass only in the old-growth forest (by 31%), but significantly elevated dead fine root biomass in all the three forests (by 64% to 101%), causing decreased live fine root proportion in the old-growth and the pine forests. P addition significantly increased live fine root biomass in all three forests (by 20% to 76%). The combined N and P treatment significantly increased live fine root biomass in the two younger forests but not in the old-growth forest. These results suggest that fine root growth in all three study forests appeared to be P-limited. This was further confirmed by current status of fine root N:P ratios, APA in bulk soil, and their responses to N and P treatments. Moreover, N addition significantly increased APA only in the old-growth forest, consistent with the conclusion that the old-growth forest was more P-limited than the younger forests.

Interactive effects of nitrogen and phosphorus on soil microbial communities in a tropical forest.

Elevated nitrogen (N) deposition in humid tropical regions may exacerbate phosphorus (P) deficiency in forests on highly weathered soils. However, it is not clear how P availability affects soil microbes and soil carbon (C), or how P processes interact with N deposition in tropical forests. We examined the effects of N and P additions on soil microbes and soil C pools in a N-saturated old-growth tropical forest in southern China to test the hypotheses that (1) N and P addition will have opposing effects on soil microbial biomass and activity, (2) N and P addition will alter the composition of the microbial community, (3) the addition of N and P will have interactive effects on soil microbes and (4) addition-mediated changes in microbial communities would feed back on soil C pools. Phospholipid fatty acid (PLFA) analysis was used to quantify the soil microbial community following four treatments: Control, N addition (15 g N m(-2) yr(-1)), P addition (15 g P m(-2) yr(-1)), and N&P addition (15 g N m(-2) yr(-1) plus 15 g P m(-2) yr(-1)). These were applied from 2007 to 2011. Whereas additions of P increased soil microbial biomass, additions of N reduced soil microbial biomass. These effects, however, were transient, disappearing over longer periods. Moreover, N additions significantly increased relative abundance of fungal PLFAs and P additions significantly increased relative abundance of arbuscular mycorrhizal (AM) fungi PLFAs. Nitrogen addition had a negative effect on light fraction C, but no effect on heavy fraction C and total soil C. In contrast, P addition significantly decreased both light fraction C and total soil C. However, there were no interactions between N addition and P addition on soil microbes. Our results suggest that these nutrients are not co-limiting, and that P rather than N is limiting in this tropical forest.

Effects of experimental nitrogen and phosphorus addition on litter decomposition in an old-growth tropical forest.

The responses of litter decomposition to nitrogen (N) and phosphorus (P) additions were examined in an old-growth tropical forest in southern China to test the following hypotheses: (1) N addition would decrease litter decomposition; (2) P addition would increase litter decomposition, and (3) P addition would mitigate the inhibitive effect of N addition. Two kinds of leaf litter, Schima superba Chardn. & Champ. (S.S.) and Castanopsis chinensis Hance (C.C.), were studied using the litterbag technique. Four treatments were conducted at the following levels: control, N-addition (150 kg N ha(-1) yr(-1)), P-addition (150 kg P ha(-1) yr(-1)) and NP-addition (150 kg N ha(-1) yr(-1) plus 150 kg P ha(-1) yr(-1)). While N addition significantly decreased the decomposition of both litters, P addition significantly inhibited decomposition of C.C., but did not affect the decomposition of S.S. The negative effect of N addition on litter decomposition might be related to the high N-saturation in this old-growth tropical forest; however, the negative effect of P addition might be due to the suppression of "microbial P mining". Significant interaction between N and P addition was found on litter decomposition, which was reflected by the less negative effect in NP-addition plots than those in N-addition plots. Our results suggest that P addition may also have negative effect on litter decomposition and that P addition would mitigate the negative effect of N deposition on litter decomposition in tropical forests.

Dynamics of soil inorganic nitrogen and their responses to nitrogen additions in three subtropical forests, south China.

Three forests with different historical land-use, forest age, and species assemblages in subtropical China were selected to evaluate current soil N status and investigate the responses of soil inorganic N dynamics to monthly ammonium nitrate additions. Results showed that the mature monsoon evergreen broadleaved forest that has been protected for more than 400 years exhibited an advanced soil N status than the pine (Pinus massoniana) and pine-broadleaf mixed forests, both originated from the 1930's clear-cut and pine plantation. Mature forests had greater extractable inorganic N pool, lower N retention capacity, higher inorganic N leaching, and higher soil C/N ratios. Mineral soil extractable NH4(+)-N and NO3(-)-N concentrations were significantly increased by experimental N additions on several sampling dates, but repeated ANOVA showed that the effect was not significant over the whole year except NH4(+)-N in the mature forest. In contrast, inorganic N (both NH4(+)-N and NO3(-)-N) in soil 20-cm below the surface was significantly elevated by the N additions. From 42% to 74% of N added was retained by the upper 20 cm soils in the pine and mixed forests, while 0%-70% was retained in the mature forest. Our results suggest that land-use history, forest age and species composition were likely to be some of the important factors that determine differing forest N retention responses to elevated N deposition in the study region.