Soil bacterial and fungal communities resilience to long-term nitrogen addition in subtropical forests in China

Atmospheric nitrogen(N)deposition is predicted to increase,especially in the subtropics.However,the responses of soil microorganisms to long-term N addition at the molecular level in N-rich subtropical forests have not been clarified.A long-term nutrient addition experiment was conducted in a subtropical evergreen old-growth forest in China.The four treatments were:control,low N(50 kg N ha^(-1)a^(-1)),high N(100 kg N ha^(-1)a^(-1)),and combined N and phosphorus(P)(100 kg N ha^(-1)a^(-1)+50 kg P ha^(-1)a^(-1)).Metagenomic sequencing characterized diversity and composition of soil microbial communities and used to construct bacterial/fungal co-occurrence networks.Nutrient-treated soils were more acidic and had higher levels of dissolved organic carbon than controls.There were no significant differences in microbial diversity and community composition across treatments.The addition of nutrients increased the abundance of copiotrophic bacteria and potentially beneficial microorganisms(e.g.,Gemmatimonadetes,Chaetomium,and Aureobasidium).Low N addition increased microbiome network connectivity.Three rare fungi were identified as module hubs under nutrient addition,indicating that low abundance fungi were more sensitive to increased nutrients.The results indicate that the overall composition of microbial communities was stable but not static to long-term N addition.Our findings provide new insights that can aid predictions of the response of soil microbial communities to long-term N addition.

Nitrogen addition promotes foliar litterfall and element return in a subtropical forest,southwestern China

Nitrogen deposition has a considerable impact on biogeochemical cycling in terrestrial ecosystems.However,how litter production and element return respond to N addition remains poorly understood in nitrogen-rich subtropical regions.In this study,a 4-year nitrogen addition experiment explored its eff ects on foliar litter production and carbon,nitrogen and phosphorus in a subtropical Michelia wilsonii forest.A clear seasonal pattern in foliar litterfall was observed,regardless of nitrogen treatments,with a peak in spring and a smaller one in autumn.Foliar litter increased with increasing nitrogen but did not aff ect litter carbon concentrations and often decreased nitrogen and phosphorous concentrations.The eff ect of nitrogen addition was dependent on time(month/year).Carbon,nitrogen and phosphorous return showed similar bimodal seasonal patterns.Nitrogen addition increased carbon and nitrogen return but did not aff ect phosphorous.Our results suggest that the addition of nitrogen stimulates carbon and nutrient return via litterfall.

Management scheme influence and nitrogen addition effects on soil CO_(2),CH_(4),and N_(2)O fluxes in a Moso bamboo plantation

Background:It is still not clear whether the effects of N deposition on soil greenhouse gas(GHG)emissions are influenced by plantation management schemes.A field experiment was conducted to investigate the effects of conventional management(CM)versus intensive management(IM),in combination with simulated N deposition levels of control(ambient N deposition),30 kg N·ha^(−1)·year^(−1)(N30,ambient+30 kg N·ha^(−1)·year^(−1)),60 kg N·ha^(−1)·year^(−1)(N60,ambient+60 kg N·ha^(−1)·year^(−1)),or 90 kg N·ha^(−1)·year^(−1)(N90,ambient+90 kg N·ha^(−1)·year^(−1))on soil CO_(2),CH_(4),and N_(2)O fluxes.For this,24 plots were set up in a Moso bamboo(Phyllostachys edulis)plantation from January 2013 to December 2015.Gas samples were collected monthly from January 2015 to December 2015.Results:Compared with CM,IM significantly increased soil CO_(2) emissions and their temperature sensitivity(Q_(10))but had no significant effects on soil CH_(4) uptake or N_(2)O emissions.In the CM plots,N30 and N60 significantly increased soil CO_(2) emissions,while N60 and N90 significantly increased soil N_(2)O emissions.In the IM plots,N30 and N60 significantly increased soil CO_(2) and N_(2)O emissions,while N60 and N90 significantly decreased soil CH_(4) uptake.Overall,in both CM and IM plots,N30 and N60 significantly increased global warming potentials,whereas N90 did not significantly affect global warming potential.However,N addition significantly decreased the Q_(10) value of soil CO_(2) emissions under IM but not under CM.Soil microbial biomass carbon was significantly and positively correlated with soil CO_(2) and N_(2)O emissions but significantly and negatively correlated with soil CH_(4) uptake.Conclusion:Our results indicate that management scheme effects should be considered when assessing the effect of atmospheric N deposition on GHG emissions in bamboo plantations.

The effect of soil moisture on the response by fungi and bacteria to nitrogen additions for N_(2)O production

In addition to bacteria,the contribution of fungi to nitrous oxide(N_(2)O)production has been recognized but the responses of these two broad and unrelated groups of microorganisms to global environmental changes,atmospheric nitrogen(N)deposition,and precipitation in terms of N_(2)O production are unclear.We studied how these two microbial-mediated N_(2)O production pathways responded to soil moisture conditions and to N addition in an N-limited temperate forest.Soils from a long-term N addition experiment in Changbai Mountain,northeastern China were incubated.Varied concentrations of cycloheximide and streptomycin,both inhibitors of fungal and bacterial activity,were used to determine the contributions of both to N_(2)O production in 66%,98%and 130%water-filled pore spaces(WFPS).The results showed that N_(2)O production decreased significantly with increasing cycloheximide concentration whereas streptomycin was only inhibiting N_(2)O emissions at 98%and 130%WFPS.The bacterial pathway of N_(2)O production in N-addition(Nadd)soil was significantly more dominant than that in untreated(Namb)soil.The difference in the fungal pathway of N_(2)O production between the soil with nitrogen addition and the untreated soil was not significant.Net N_(2)O emissions increased with increasing soil moisture,especially at 130%WFPS,a completely flooded condition.Bacteria dominated carbon dioxide(CO_(2))and N_(2)O emissions in Nadd soil and at 130%WFPS regardless of N status,while fungi dominated CO_(2)and N_(2)O emissions in soil without N addition at 66%and 98%WFPS.The results suggest that flooded soil is an important source of N_(2)O emissions and that bacteria might be better adapted to compete in fertile soils under anoxic conditions.

Reproductive height determines the loss of clonal grasses with nitrogen enrichment in a temperate grassland

Tall clonal grasses commonly display competitive advantages with nitrogen(N)enrichment.However,it is currently unknown whether the height is derived from the vegetative or reproductive module.Moreover,it is unclear whether the height of the vegetative or reproductive system regulates the probability of extinction and colonization,and determines species diversity.In this study,the impacts on clonal grasses were studied in a field experiment employing two frequencies(twice a year vs.monthly)crossing with nine N addition rates in a temperate grassland,China.We found that the N addition decreased species frequency and increased extinction probability,but did not change the species colonization probability.A low frequency of N addition decreased species frequency and colonization probability,but increased extinction probability.Moreover,we found that species reproductive height was the best index to predict the extinction probability of clonal grasses in N-enriched conditions.The low frequency of N addition may overestimate the negative effect from N deposition on clonal grass diversity,suggesting that a higher frequency of N addition is more suitable in assessing the ecological effects of N deposition.Overall,this study illustrates that reproductive height was associated with the clonal species extinction probability under N-enriched environment.

Effects of nitrogen and phosphorus additions on soil microbial community structure and ecological processes in the farmland of Chinese Loess Plateau

Microorganisms regulate the responses of terrestrial ecosystems to anthropogenic nutrient inputs.The escalation of anthropogenic activities has resulted in a rise in the primary terrestrial constraining elements,namely nitrogen(N)and phosphorus(P).Nevertheless,the specific mechanisms governing the influence of soil microbial community structure and ecological processes in ecologically vulnerable and delicate semi-arid loess agroecosystems remain inadequately understood.Therefore,we explored the effects of different N and P additions on soil microbial community structure and its associated ecological processes in the farmland of Chinese Loess Plateau based on a 36-a long-term experiment.Nine fertilization treatments with complete interactions of high,medium,and low N and P gradients were set up.Soil physical and chemical properties,along with the microbial community structure were measured in this study.Additionally,relevant ecological processes such as microbial biomass,respiration,N mineralization,and enzyme activity were quantified.To elucidate the relationships between these variables,we examined correlation-mediated processes using statistical techniques,including redundancy analysis(RDA)and structural equation modeling(SEM).The results showed that the addition of N alone had a detrimental effect on soil microbial biomass,mineralized N accumulation,andβ-1,4-glucosidase activity.Conversely,the addition of P exhibited an opposing effect,leading to positive influences on these soil parameters.The interactive addition of N and P significantly changed the microbial community structure,increasing microbial activity(microbial biomass and soil respiration),but decreasing the accumulation of mineralized N.Among them,N24P12 treatment showed the greatest increase in the soil nutrient content and respiration.N12P12 treatment increased the overall enzyme activity and total phospholipid fatty acid(PLFA)content by 70.93%.N and P nutrient contents of the soil dominate the microbial community structure and the corresponding changes in hydrolytic enzymes.Soil microbial biomass,respiration,and overall enzyme activity are driven by mineralized N.Our study provides a theoretical basis for exploring energy conversion processes of soil microbial community and environmental sustainability under long-term N and P additions in semi-arid loess areas.

Effects of mowing and nitrogen addition on soil respiration in three patches in an oldfield grassland in Inner Mongolia

Aims Vegetation type is important in determining variations in soil carbon(C)efflux under grassland managements.This study was conducted to examine the effects of mowing and nitrogen(N)addition on soil respiration and their dependences upon vegetation types in an oldfield grassland of northern China.Methods Soil respiration,temperature,moisture and aboveground net primary productivity(ANPP)and belowground net primary productivity(BNPP)were examined in response to mowing and N addition among the three patches dominated by different species(named as grass,forb and mixed patches,respectively)in the growing seasons(May–October)from 2006 to 2008.Important Findings Across the 3 years,soil respiration in the grass patch was greater than those in the forb and mixed patches,which could have been ascribed to the higher soil moisture(SM)in the grass patch.Mowing had no impact on soil respiration due to unaltered SM and plant growth.Soil respiration was stimulated by 6.53%under N addition,and the enhancement was statistically significant in 2006 but not in 2007 or 2008 because of the limited water availability in the later 2 years.There were no interactive effects between mowing and N addition on soil respiration.Soil respiration showed positive dependence upon SM,ANPP and BNPP across plots.The results suggest that soil water availability and plant growth could be the primary factors in controlling the temporal and spatial variations in soil respiration and its response to different treatments.Our observations indicate that grassland managements(i.e.mowing for hay once a year)may have little influence on soil respiration of the oldfield grassland in northern China.

comLinking flowering and reproductive allocation in response to nitrogen addition in an alpine meadow

Aims Plants can change in phenology and biomass allocation in response to environmental change.It has been demonstrated that nitrogen is the most limiting resource for plants in many terrestrial ecosystems.Previous studies have usually focused on either flowering phenology or biomass allocation of plants in response to nitrogen addition;how-ever,attempts to link flowering phenology and biomass allocation are still rare.In this study,we tested the effects of nitrogen addition on both flowering phenology and reproductive allocation in 34 common species.We also examined the potential linkage between flowering time and reproductive allocation in response to nitrogen addition.Methods We conducted a 3-year nitrogen addition experiment in Tibetan alpine meadow.We measured first flowering date and the repro-ductive allocation for 34 common plant species in control,low and high nitrogen added plots,respectively.one-way analysis of variance was used to examine differences of first flowering date and reproductive allocation among treatments.The relationships between the change in species first flowering date and change in reproductive allocation in response to nitrogen addition were examined by calculating Pearson correlation coefficients.Important Findings For most species,both first flowering date and reproductive alloca-tion significantly responded to nitrogen addition.Nitrogen addition significantly delayed the first flowering date and reduced the repro-ductive allocation for all graminoid species,but accelerated flower-ing and increased reproductive allocation for most forb species.We found that changes in first flowering date significantly negatively correlated with the changes in reproductive allocation over spe-cies in response to nitrogen,which indicated a positive relationship between flowering response and plant performance in reproductive allocation.species that advanced their flowering time with nitrogen addition increased their reproductive allocation,whereas those that delayed flowering time tended to decline in reproductive allocation with nitrogen addition.our results suggest that species-specific switch from vegetative growth to reproductive growth could influence species performance.

Effects of nitrogen addition on plant biomass and tissue elemental content in different degradation stages of temperate steppe in northern China

Aims Grassland degradation has become a common problem worldwide.Several studies have analyzed the effects of nitrogen(N)addition on plant growth in grasslands,but few have considered its effects on plant growth in degraded grasslands.The aim of this study was to evaluate the effects of N addition on plant growth in grasslands with different levels of degradation in Inner Mongolia,China.Methods A 2-year field experiment was conducted to evaluate plant growth response to N addition in degraded grasslands on the Inner Mongolian Steppe.Grasslands with four levels of degradation were selected for N-addition treatments(0,10,20,30,40,50 g N m−2 year−1).Important Findings Aboveground biomass was increased by N fertilization in degraded grasslands,and N addition was significantly related to increased biomass in grasslands with severe degradation.However,N fertilization did not significantly affect belowground biomass.The effects of N addition on foliar nutrient concentrations in the same species differed among grasslands with different degradation lev-els.There was an inconsistent response to N addition between Sanguisorba officinalis and Vicia sepium in non-degraded grassland.There was a significant positive correlation between foliar N content and aboveground biomass in grasslands with different levels of deg-radation.Our results indicate that the effects of N addition on plant growth in grasslands differ according to the severity of degradation.We conclude that N fertilization may be an effective management technique for degraded grasslands in this area and may improve for-age productivity in the short term.

Nitrogen addition frequency and propagule pressure influence Solidago canadensis invasion into native plant community

Introduction:Propagule pressure(i.e.,the number of propagules)has long been recognized to play an essential role in plant invasion.But it is not clear whether propagule pressure influences the invasion of exotic plants into native plant communities when different frequencies of nitrogen are added.Method:We established an experiment with three plant communities that included native plant communities alone(four grasses,two legumes and two forbs)or native plant communities with one or five invasive plants,Solidago canadensis,under three frequencies of nitrogen addition(no addition or low or high addition with the same amount).Results:High propagule pressure significantly enhanced the biomass and relative dominance index of S.canadensis.Moreover,high propagule pressure only decreased the total and aboveground biomass of the legumes.However,the competitive effect between S.canadensis and the native community and biomass of the whole native community varied according to different frequencies.Conclusion:Overall,high propagule pressure encouraged invasion by S.canadensis,while alow nitrogen frequency was advantageous for the native community to resist invasion in this experiment.The results provide a scientific basis to manage and control the invasion of S.canadensis.

Biochar amendments increase soil organic carbon storage and decrease global warming potentials of soil CH4 and N2O under N addition in a subtropical Moso bamboo plantation

Background: Nitrogen(N) deposition affects soil greenhouse gas(GHG) emissions, while biochar application reduces GHG emissions in agricultural soils. However, it remains unclear whether biochar amendment can alleviate the promoting effects of N input on GHG emissions in forest soils. Here, we quantify the separate and combined effects of biochar amendment(0, 20, and 40 t·ha) and N addition(0, 30, 60, and 90 kg N·ha·yr) on soil GHG fluxes in a long-term field experiment at a Moso bamboo(Phyllostachys edulis) plantation.Results: Low and moderate N inputs(≤60 kg N·ha·yr) significantly increase mean annual soil carbon dioxide(CO) and nitrous oxide(NO) emissions by 17.0%–25.4% and 29.8%–31.2%, respectively, while decreasing methane(CH) uptake by 12.4%–15.9%, leading to increases in the global warming potential(GWP) of soil CHand NO fluxes by 32.4%–44.0%. Moreover, N addition reduces soil organic carbon(C;SOC) storage by 0.2%–6.5%. Compared to the control treatment, biochar amendment increases mean annual soil CO2emissions, CHuptake, and SOC storage by 18.4%–25.4%, 7.6%–15.8%, and 7.1%–13.4%, respectively, while decreasing NO emissions by 17.6%–19.2%, leading to a GWP decrease of 18.4%–21.4%. Biochar amendments significantly enhance the promoting effects of N addition on soil COemissions, while substantially offsetting the promotion of N2O emissions, inhibition of CHuptake, and decreased SOC storage, resulting in a GWP decrease of 9.1%–30.3%.Additionally, soil COand CHfluxes are significantly and positively correlated with soil microbial biomass C(MBC) and pH. Meanwhile, NO emissions have a significant and positive correlation with soil MBC and a negative correlation with pH.Conclusions: Biochar amendment can increase SOC storage and offset the enhanced GWP mediated by elevated N deposition and is, thus, a potential strategy for increasing soil C sinks and decreasing GWPs of soil CHand NO under increasing atmospheric N deposition in Moso bamboo plantations.

Warming intensified the effects of nitrogen addition on N_(2)O emissions from alpine meadow in the northern Qinghai-Tibet Plateau

Warming and nitrogen(N)addition may impact soil nitrous oxide(N_(2)O)emissions,but the relationship between plant community composition and soil microbial activities remains unclear.For a two-year field study in the Qinghai-Tibet Plateau,open-top chambers were used to quantify the effects of warming,N-addition,and their interactions on N_(2)O emissions.We found that the N-addition greatly increased N_(2)O emissions by 77.4%in 2018 when compared to the control group.In contrast,warming showed little effect on N_(2)0 emissions but did increase the activity of enzymes associated with soil nitrification and denitrification.A combined effect of warming and N-addition of resulted in 208.6%(2018)and 90.8%(2019)increase in N_(2)0 emissions,respectively,compared to the individual treatments of warming or N-addition.Global warming in alpine meadows is causally linked to increased legume biomass which is further intensified with the N-addition.Intensified legume biomass(p<0.05),soil moisture(p<0.001)and enzyme activity(p<0.001)had a positive effect on N_(2)0 emissions,while diminished microbial carbon/nitrogen(MBC/MBN)(p<0.05)correlated with reduced N_(2)O emissions.Final results indicated that N-addition has a positive effect on N_(2)O emissions,and the addition of warming further intensifies this effect.The increased dominance of legumes and microbial N content contributes to this effect.These outcomes suggest that warming and atmospheric N deposition can stimulate N_(2)O emissions of alpine meadows inthefuture.

Nutrient coordination mechanism of tiger nut induced by rhizosphere soil nutrient variation in an arid area, China

Tiger nut is a bioenergy crop planted in arid areas of northern China to supply oil and adjust the planting structure.However,in the western region of Inner Mongolia Autonomous Region,China,less water resources have resulted in a scarcity of available farmland,which has posed a huge obstacle to planting tiger nut.Cultivation of tiger nut on marginal land can effectively solve this problem.To fully unlock the production potential of tiger nut on marginal land,it is crucial for managers to have comprehensive information on the adaptive mechanism and nutrient requirement of tiger nut in different growth periods.This study aims to explore these key information from the perspective of nutrient coordination strategy of tiger nut in different growth periods and their relationship with rhizosphere soil nutrients.Three fertilization treatments including no fertilization(N:P(nitrogen:phosphorous)=0:0),traditional fertilization(N:P=15:15),and additional N fertilizer(N:P=60:15)were implemented on marginal land in the Dengkou County.Plant and soil samples were collected in three growth periods,including stolon tillering period,tuber expanding period,and tuber mature period.Under no fertilization,there was a significant correlation between N and P contents of tiger nut roots and tubers and the same nutrients in the rhizosphere soil(P<0.05).Carbon(C),N,and P contents of roots were significantly higher than those of leaves(P<0.05),and the C:N ratio of all organs was higher than those under other treatments before tuber maturity(P<0.05).Under traditional fertilization,there was a significant impact on the P content of tiger nut tubers(P<0.05).Under additional N fertilizer,the accumulation rate of N and P was faster in stolons than in tubers(P<0.05)with lower N:P ratio in stolons during the tuber expansion period(P<0.05),but higher N:P ratio in tubers(P<0.05).The limited availability of nutrients in the rhizosphere soil prompts tiger nut to increase the C:N ratio,improving N utilization efficiency,and maintaining N:P ratio in tubers.Elevated N levels in the rhizosphere soil decrease the C:N ratio of tiger nut organs and N:P ratio in stolons,promoting rapid stolon growth and shoot production.Supplementary P is necessary during tuber expansion,while a higher proportion of N in fertilizers is crucial for the aboveground biomass production of tiger nut.

Autumn nitrogen enrichment destabilizes ecosystem biomass production in a semiarid grassland

Nitrogen(N)deposition decreases the temporal stability of ecosystem aboveground biomass production(ecosystem stability).However,little is known about how the responses of ecosystem stability differ based on seasonal N enrichment.By adding N in autumn,winter,or growing season,from October 2014 to May 2020,in a temperate grassland in northern China,we found that only N addition in autumn resulted in a significantly positive correlation between ecosystem mean aboveground net primary productivity(ANPP)and its standard deviation and significantly reduced ecosystem stability.Autumn N-induced reduction in ecosystem stability was associated with the vanished negative effect of community-wide species asynchrony(asynchronous dynamics among populations to environmental perturbations)on the standard deviation of ecosystem ANPP in combination with the emerged positive effect of dominance(Simpson's dominance index that indicates the relative weight of dominant species in a community).Our findings indicate that autumn N addition might overestimate the negative effect of annual atmospheric N deposition on ecosystem stability,suggesting that to better evaluate the influence of N deposition in temperate grasslands,both field experiments and global modeling should consider not only the annual N load but also its seasonal dynamics.Moreover,further studies should pay more attention to the alteration in the ecosystem temporal deviations,which might be more sensitive to human-induced environmental changes.

玉米氮肥适宜追施期研究

玉米在大喇叭口期重施氮肥，植株矮，穗位低，利于抗倒伏。中后期叶片衰亡慢，比叶重高，干物质积累多，从而果穗秃尖短，穗粒数多，千粒重高。大喇叭口期施肥较３叶期施肥增产１５７％，比拔节期施肥增产１０．６％，比吐丝期施肥增产６．８％。拔节期和大喇叭口期分两次追施和大喇叭口期一次施入产量相近。大喇叭口期是玉米最适宜的追肥期。

Divergent responses of plant biomass and diversity to short-term nitrogen and phosphorus addition in three types of steppe in Inner Mongolia,China

Background:Understanding the response of the plant community to increasing nitrogen(N)and phosphorus(P)inputs is helpful for managing and protecting grassland ecosystems in semiarid areas.However,information about different types of steppe responses to N and P availability in semiarid grasslands is limited.In 2017-2018,two field experiments were conducted with six levels of N(from 5 to 30 g N m^(−2)yr^(−1))and P(from 2.5 g to 15 g P m^(−2)yr^(−1))additions in three different temperate steppes,including meadow steppe(MS),typical steppe(TS),and desert steppe(DS),in northern China to study the effects of these addition rates on community biomass and diversity.Results:Our results showed that plant biomass and diversity in the three steppe types in Inner Mongolia responded differently to elevated N and P inputs.Increasing P promoted aboveground and belowground biomass more than increasing N in the three temperate steppes.Short-term N and P additions reduced plant diversity to some extent,with the most pronounced decreases in MS and DS.It is noteworthy that there were response thresholds for plant diversity and biomass in response to N and P inputs in different steppe types(e.g.,10 g P m^(−2)yr^(−1)).Furthermore,redundancy analysis and stepwise regression analysis revealed that changes in soil properties induced by nutrient addition and climate conditions jointly regulated changes in vegetation biomass and diversity.Conclusions:The plant biomass and diversity of three steppe types in Inner Mongolia respond divergently to elevated N and P inputs.Our results indicate that regional differences in climate and soil substrate conditions may jointly contribute to the divergent responses of plant biomass and diversity to short-term N and P addition.Our analyses provide new insights into managing and protecting grassland ecosystems.Considering that the effects of nutrient addition on plant diversity and productivity may have increasing effects over time,studies on long-term in situ nutrient addition are necessary.

Impacts of nitrogen deposition on terrestrial plant diversity: a meta-analysis in China

Aims With the global atmospheric nitrogen(N)deposition increasing,the effect of N deposition on terrestrial plant diversity has been widely studied.Some studies have reviewed the effects of N deposition on plant species diversity;however,all studies addressed the effects of N deposition on plant community focused on species richness in specific ecosystem.There is a need for a systematic meta-analysis covering multiple dimensions of plant diversity in multiple climate zones and ecosystems types.Our goal was to quantify changes in species richness,evenness and uncertainty in plant communities in response to N addition across different environmental and experimental contexts.Methods We performed a meta-analysis of 623 experimental records published in English and Chinese journals to evaluate the response of terrestrial plant diversity to the experimental N addition in China.Three metrics were used to quantify the change in plant diversity:species richness(SR),evenness(Pielou index)uncertainty(Shannon index).Important Findings Results showed that(i)N addition negatively affected SR in temperate,Plateau zones and subtropical zone,but had no significant effect on Shannon index in subtropical zones;(ii)N addition decreased SR,Shannon index and Pielou index in grassland,and the negative effect of N addition on SR was stronger in forest than in grassland;(iii)N addition negatively affected plant diversity(SR,Shannon index and Pielou index)in the long term,whereas it did not affect plant diversity in the short term.Furthermore,the increase in N addition levels strengthened the negative effect of N deposition on plant diversity with long experiment duration;and(iv)the negative effect of ammonium nitrate(NH_(4)NO_(3))addition on SR was stronger than that of urea(CO(NH_(2))_(2))addition,but the negative effect of NH_(4)NO_(3) addition on Pielou index was weaker than that of CO(NH2)2 addition.Our results indicated that the effects of N addition on plant diversity varied depending on climate zones,ecosystem types,N addition levels,N type and experiment duration.This underlines the importance of integrating multiple dimensions of plant diversity and multiple factors into assessments of plant diversity to global environmental change.

Effects of the frequency and the rate of N enrichment on community structure in a temperate grassland

Aims Nitrogen(N)enrichment caused by human activities threatens bio-diversity and alters plant community composition and structure.It has been found that heavy and infrequent N inputs may over-estimate species extinction,but it remains unclear whether plant community structure will equally respond to frequent reactive N enriched conditions.Methods We independently manipulated the rates and the frequencies of N addition in a temperate steppe,northern China,between 2008 and 2013.Important Findings We found that plant community structure changes,measured by‘Euclidean distance’involving species richness,composition and productivity,were significantly positively related to increasing N enrichment rates rather than frequencies.Changes in aboveground net primary productivity(ANPP),plant species richness and shifts in dominant species were observed.Community ANPP increased with N enrichment,whereas species richness reduced.The frequency of N enrichment increased species richness but had no impacts on community ANPP and the relative ANPP of the two dominant spe-cies,C3 perennial bunchgrass Stipa grandis and C3 perennial rhi-zome grass Leymus chinensis.The ANPP and relative ANPP of the two dominant species were significantly negatively correlated with each other.Moreover,changes in the relative ANPP of S.grandis was negatively associated with the changes in community structure.After 5 years’treatment,direct influence of the frequency of N en-richment on plant community structure was not observed,but the effects of the rate of N enrichment were apparent.Our results sug-gested that further study in various ecosystems and with long-term and well-controlled comparisons the frequency vs.the rate of N enrichment may still be needed.