Effect of life cycle and venation pattern on the coordination between stomatal and vein densities of herbs.

Life cycle (annual vs perennial) and leaf venation pattern (parallel and reticular) are known to be related to water use strategies in herb species and critical adaptation to certain climatic conditions. However, the effect of these two traits and how they influence the coordination between vein density (vein length per area, VLA) and stomatal density (SD) remains unclear. In this study, we examined the leaves of 53 herb species from a subtropical botanical garden in Guangdong Province, China, including herbs with different life cycles and leaf venation patterns. We assessed 21 leaf water-related functional traits for all species, including leaf area (LA), major and minor VLA, major and minor vein diameter (VD), SD and stomatal length (SL). The results showed no significant differences in mean SD and SL between either functional group (parallel venation vs reticular venation and annual vs perennial). However, parallel vein herbs and perennial herbs displayed a significantly higher mean LA and minor VD, and lower minor VLA compared to reticular vein herbs and annual herbs, respectively. There was a linear correlation between total VLA and SD in perennial and reticular vein herbs, but this kind of correlation was not found in annual and parallel vein herbs. The major VLA and minor VD were significantly affected by the interaction between life cycle and leaf venation pattern. Our findings suggested that VLA, rather than SD, may serve as a more adaptable structure regulated by herbaceous plants to support the coordination between leaf water supply and demand in the context of different life cycles and leaf venation patterns. The results of the present study provide mechanistic understandings of functional advantages of different leaf types, which may involve in species fitness in community assembly and divergent responses to climate changes.

Effect of leaf phenology and morphology on the coordination between stomatal and minor vein densities.

Leaf phenology (evergreen vs. deciduous) and morphology (simple vs. compound) are known to be related to water use strategies in tree species and critical adaptation to certain climatic conditions. However, the effect of these two traits and their interactions on the coordination between minor vein density (MVD) and stomatal density (SD) remains unclear. In this study, we examined the leaves of 108 tree species from plots in a primary subtropical forest in southern China, including tree species with different leaf morphologies and phenologies. We assessed nine leaf water-related functional traits for all species, including MVD, SD, leaf area (LA), minor vein thickness (MVT), and stomatal length (SL). The results showed no significant differences in mean LA and SD between either functional group (simple vs. compound and evergreen vs. deciduous). However, deciduous trees displayed a significantly higher mean MVD compared to evergreen trees. Similarly, compound-leaved trees have a higher (marginally significant) MVD than simple-leaved trees. Furthermore, we found that leaf morphology and phenology have significantly interactive effects on SL, and the compound-leafed deciduous trees exhibited the largest average SL among the four groups. There were significant correlations between the MVD and SD in all different tree groups; however, the slopes and interceptions differed within both morphology and phenology. Our results indicate that MVD, rather than SD, may be the more flexible structure for supporting the coordination between leaf water supply and demand in different leaf morphologies and phenologies. The results of the present study provide mechanistic understandings of the functional advantages of different leaf types, which may involve species fitness in community assembly and divergent responses to climate changes.

Patterns of compound-leaf form and deciduous-leaf habit across forests in China: Their association and key climatic factors.

Leaf form (compound vs. simple) and habit (evergreen vs. deciduous) are key functional traits of trees to adapt to various climates and are vital in determining plant response to climate change. However, their association and climatic determinants remain uncertain, especially in East Asian forests in the largest monsoon region on earth. To fill these knowledge gaps, we compiled a dataset comprising 42 intact forests and over 2200 angiosperm tree species across China (spanning 30 latitudes and 47 longitudes). The geographical and climatic patterns of leaf form and habit were analyzed. The association between compound leaf and deciduousness was tested for tropical, subtropical and temperate climatic zones. We found that both the percentage of compound leaf (CT%) and deciduous tree species (DT%) increased with latitude and decreased with mean annual precipitation (MAP). For all forests, DT% was negatively related to mean annual temperature (MAT), whereas CT% was not. Nevertheless, both DT% and CT% increased with increasing MAT in the tropics, possibly owing to the high vapor pressure deficits (VPD) and canopy water deficits associated with high temperatures. A positive linear relationship between CT% and DT% was found across all forests and within different climatic zones except for temperate, and the intercept of the regression line was significantly higher in the tropics than in the subtropics. Overall, as supported by principal component analysis, deciduousness was negatively associated with both temperature and precipitation, while CT negatively with precipitation only across zones and positively with temperature in the tropics. Different relationships in different climatic zones suggest potentially different selective forces. Our findings provide novel insights into the linkage between leaf form and habit, as well as how climate shapes the landscape of broadleaf forests, which has important implications regarding the response of forest composition to climate change.

Unexpected high retention of 15 N-labeled nitrogen in a tropical legume forest under long-term nitrogen enrichment.

The responses of forests to nitrogen (N) deposition largely depend on the fates of deposited N within the ecosystem. Nitrogen-fixing legume trees widely occur in terrestrial forests, but the fates of deposited N in legume-dominated forests remain unclear, which limit a global evaluation of N deposition impacts and feedbacks on carbon sequestration. Here, we performed the first ecosystem-scale 15 N labeling experiment in a typical legume-dominated forest as well as in a nearby non-legume forest to determine the fates of N deposition between two different forest types and to explore their underlying mechanisms. The 15 N was sprayed bimonthly for 1 year to the forest floor in control and N addition (50 kg N ha-1  year-1 for 10 years) plots in both forests. We unexpectedly found a strong capacity of the legume forest to retain deposited N, with 75 ± 5% labeled N recovered in plants and soils, which was higher than that in the non-legume forest (56 ± 4%). The higher 15 N recovery in legume forest was mainly driven by uptake by the legume trees, in which 15 N recovery was approximately 15% more than that in the nearby non-legume trees. This indicates higher N-demand by the legume than non-legume trees. Mineral soil was the major sink for deposited N, with 39 ± 4% and 34 ± 3% labeled N retained in the legume and non-legume forests, respectively. Moreover, N addition did not significantly change the 15 N recovery patterns of both forests. Overall, these findings indicate that legume-dominated forests act as a strong sink for deposited N regardless of high soil N availability under long-term atmospheric N deposition, which suggest a necessity to incorporate legume-dominated forests into N-cycling models of Earth systems to improve the understanding and prediction of terrestrial N budgets and the global N deposition effects.

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.

Nitrogen deposition accelerates soil carbon sequestration in tropical forests.

Terrestrial ecosystem carbon (C) sequestration plays an important role in ameliorating global climate change. While tropical forests exert a disproportionately large influence on global C cycling, there remains an open question on changes in below-ground soil C stocks with global increases in nitrogen (N) deposition, because N supply often does not constrain the growth of tropical forests. We quantified soil C sequestration through more than a decade of continuous N addition experiment in an N-rich primary tropical forest. Results showed that long-term N additions increased soil C stocks by 7 to 21%, mainly arising from decreased C output fluxes and physical protection mechanisms without changes in the chemical composition of organic matter. A meta-analysis further verified that soil C sequestration induced by excess N inputs is a general phenomenon in tropical forests. Notably, soil N sequestration can keep pace with soil C, based on consistent C/N ratios under N additions. These findings provide empirical evidence that below-ground C sequestration can be stimulated in mature tropical forests under excess N deposition, which has important implications for predicting future terrestrial sinks for both elevated anthropogenic CO2 and N deposition. We further developed a conceptual model hypothesis depicting how soil C sequestration happens under chronic N deposition in N-limited and N-rich ecosystems, suggesting a direction to incorporate N deposition and N cycling into terrestrial C cycle models to improve the predictability on C sink strength as enhanced N deposition spreads from temperate into tropical systems.

Resource limitation of soil microbes in karst ecosystems.

Knowledge about resource limitation to soil microbes is crucial for understanding ecosystem functions and processes, and for predicting ecosystem responses to global changes as well. Karst ecosystems are widespread in the world, and play a key role in regulating the global climate, however, the patterns of and mechanisms underlying microbial resource limitation in karst ecosystems remain poorly known. Here we investigated the microbial resource limitation in a karst region, by selecting four main land-use types, i.e. cropland, grassland, shrubland and secondary forest, in areas underlain by two lithology types, i.e. dolomite and limestone, in southwest China. Ecoenzymatic stoichiometry was used as an indicator of microbial resource limitation. Overall, soil microbes in karst ecosystems were more limited by carbon and phosphorus, rather than by nitrogen. Further analyses revealed that the patterns of carbon and phosphorus limitation were different among land-use or lithology types. Microbial carbon limitation was greatest in cropland and forest but lowest in grassland, and was greater under dolomite than under limestone. Microbial phosphorus limitation decreased from secondary forest to cropland under dolomite areas, but showed no difference among ecosystem types under limestone areas, indicating that lithology controls the pattern of microbial phosphorus limitation along the post-agriculture succession. Our study describes a general pattern of microbial resource limitation in karst ecosystems, and we suggest that lithology may provide a new mechanism for explaining the variations of microbial resource limitation along the post-agriculture succession in different regions.

Plant acclimation to long-term high nitrogen deposition in an N-rich tropical forest.

Anthropogenic nitrogen (N) deposition has accelerated terrestrial N cycling at regional and global scales, causing nutrient imbalance in many natural and seminatural ecosystems. How added N affects ecosystems where N is already abundant, and how plants acclimate to chronic N deposition in such circumstances, remains poorly understood. Here, we conducted an experiment employing a decade of N additions to examine ecosystem responses and plant acclimation to added N in an N-rich tropical forest. We found that N additions accelerated soil acidification and reduced biologically available cations (especially Ca and Mg) in soils, but plants maintained foliar nutrient supply at least in part by increasing transpiration while decreasing soil water leaching below the rooting zone. We suggest a hypothesis that cation-deficient plants can adjust to elevated N deposition by increasing transpiration and thereby maintaining nutrient balance. This result suggests that long-term elevated N deposition can alter hydrological cycling in N-rich forest ecosystems.

Effects of simulated N deposition on foliar nutrient status, N metabolism and photosynthetic capacity of three dominant understory plant species in a mature tropical forest.

Anthropogenic increase of nitrogen (N) deposition has threatened forest ecosystem health at both regional and global scales. In N-limited ecosystems, atmospheric N input is regarded as an important nutrient source for plant growth. However, it remains an open question on how elevated N deposition affects plant growth in N-rich forest ecosystems. To address this question, we used a simulated N deposition experiment in an N-rich mature tropical forest of southern China, with N addition levels as 0kgNha-1yr-1 (Control), 50kgNha-1yr-1 (Low-N), 100kgNha-1yr-1 (Middle-N) and 150kgNha-1yr-1 (High-N), respectively. We measured foliar nutrient element status (e.g., N, P, K, Ca and Mg), N metabolism and photosynthesis capacity of three dominant understory plant species (Cryptocarya concinna and Cryptocarya chinensis as medium-light species; and Randia canthioides as shade tolerant species) in this forest. Results showed that two years of N addition greatly increased foliar N content, but decreased the content of nutrient cations (e.g., K, Ca and Mg). Nitrogen addition also increased N accumulation as organic forms as soluble protein and/or free amino acid (FAA), but not as chlorophyll in all three species. We further found that the photosynthesis capacity (Pmax) of C. concinna and C. chinensis decreased significantly with elevated N addition, with no effects on R. canthioides. However, photosynthetic nitrogen use efficiency (PNUE) significantly declined with N addition for all three species, with significantly negative relationships between PNUE/Pmax and foliar N content. These findings suggest that excess N inputs can accelerate nutrient imbalance, and inhibit photosynthetic capacity of understory plant species, indicating continuous high N deposition can threat understory plant growth in N-rich tropical forests in the future. Meanwhile, PNUE can be used as a sensitive indicator to assess ecosystem N status under chronic N deposition.

Different responses of asymbiotic nitrogen fixation to nitrogen addition between disturbed and rehabilitated subtropical forests.

Asymbiotic nitrogen (N) fixation is an important source of new N in ecosystems, and is sensitive to atmospheric N deposition. However, there is limited understanding of asymbiotic N fixation and its response to N deposition in the context of forest rehabilitation. In this study, we measured N fixation rates (acetylene reduction) in different ecosystem compartments (i.e. soil, forest floor, moss Syrrhopodon armatus, and canopy leaves) in a disturbed and a rehabilitated subtropical forest in southern China, under 12years of N treatments: control, low N addition (50kgNha-1yr-1), and medium N addition (100kgNha-1yr-1). The rehabilitated forest had higher nutrient (e.g. N) availability than the disturbed forest. In control plots, N fixation rates in forest floor were higher in the rehabilitated forest than in the disturbed forest, but N fixation rates in other compartments (soil, S. armatus, and canopy leaves) were comparable between the forests. Nitrogen addition significantly suppressed N fixation in soil, forest floor, S. armatus, and canopy leaves in the disturbed forest, but had no significant effect on those compartments in the rehabilitated forest. The main reasons for the negative effects of N addition on N fixation in the disturbed forest were NH4+ inhibition (soil), the P and C limitation (forest floor), and the reduced N dependence on canopy N-fixers (S. armatus and canopy leaves). We conclude that asymbiotic N fixation does not decline with increasing N availability after rehabilitation in the study forests. The inhibitory effects of N addition on asymbiotic N fixation occurred in the disturbed forest but not in the rehabilitated forest, indicating that forest rehabilitation may change the response of ecosystem function (i.e. N fixation) to N deposition, which merits further study in other tropical and subtropical regions.

High retention of 15 N-labeled nitrogen deposition in a nitrogen saturated old-growth tropical forest.

The effects of increased reactive nitrogen (N) deposition in forests depend largely on its fate in the ecosystems. However, our knowledge on the fates of deposited N in tropical forest ecosystems and its retention mechanisms is limited. Here, we report the results from the first whole ecosystem 15 N labeling experiment performed in a N-rich old-growth tropical forest in southern China. We added 15 N tracer monthly as 15 NH415 NO3 for 1 year to control plots and to N-fertilized plots (N-plots, receiving additions of 50 kg N ha-1  yr-1 for 10 years). Tracer recoveries in major ecosystem compartments were quantified 4 months after the last addition. Tracer recoveries in soil solution were monitored monthly to quantify leaching losses. Total tracer recovery in plant and soil (N retention) in the control plots was 72% and similar to those observed in temperate forests. The retention decreased to 52% in the N-plots. Soil was the dominant sink, retaining 37% and 28% of the labeled N input in the control and N-plots, respectively. Leaching below 20 cm was 50 kg N ha-1  yr-1 in the control plots and was close to the N input (51 kg N ha-1  yr-1 ), indicating N saturation of the top soil. Nitrogen addition increased N leaching to 73 kg N ha-1  yr-1 . However, of these only 7 and 23 kg N ha-1  yr-1 in the control and N-plots, respectively, originated from the labeled N input. Our findings indicate that deposited N, like in temperate forests, is largely incorporated into plant and soil pools in the short term, although the forest is N-saturated, but high cycling rates may later release the N for leaching and/or gaseous loss. Thus, N cycling rates rather than short-term N retention represent the main difference between temperate forests and the studied tropical forest.

Divergent responses of soil buffering capacity to long-term N deposition in three typical tropical forests with different land-use history.

Elevated anthropogenic nitrogen (N) deposition has become an important driver of soil acidification at both regional and global scales. It remains unclear, however, how long-term N deposition affects soil buffering capacity in tropical forest ecosystems and in ecosystems of contrasting land-use history. Here, we expand on a long-term N deposition experiment in three tropical forests that vary in land-use history (primary, secondary, and planted forests) in Southern China, with N addition as NH4NO3 of 0, 50, 100, and 150 kg N ha(-1) yr(-1), respectively. Results showed that all three forests were acid-sensitive ecosystems with poor soil buffering capacity, while the primary forest had higher base saturation and cation exchange capacity than others. However, long-term N addition significantly accelerated soil acidification and decreased soil buffering capacity in the primary forest, but not in the degraded secondary and planted forests. We suggest that ecosystem N status, influenced by different land-use history, is primarily responsible for these divergent responses. N-rich primary forests may be more sensitive to external N inputs than others with low N status, and should be given more attention under global changes in the future, because lack of nutrient cations is irreversible.

Nitrogen deposition contributes to soil acidification in tropical ecosystems.

Elevated anthropogenic nitrogen (N) deposition has greatly altered terrestrial ecosystem functioning, threatening ecosystem health via acidification and eutrophication in temperate and boreal forests across the northern hemisphere. However, response of forest soil acidification to N deposition has been less studied in humid tropics compared to other forest types. This study was designed to explore impacts of long-term N deposition on soil acidification processes in tropical forests. We have established a long-term N-deposition experiment in an N-rich lowland tropical forest of Southern China since 2002 with N addition as NH4 NO3 of 0, 50, 100 and 150 kg N ha(-1)  yr(-1) . We measured soil acidification status and element leaching in soil drainage solution after 6-year N addition. Results showed that our study site has been experiencing serious soil acidification and was quite acid-sensitive showing high acidification (pH(H2O) <4.0), negative water-extracted acid neutralizing capacity (ANC) and low base saturation (BS,< 8%) throughout soil profiles. Long-term N addition significantly accelerated soil acidification, leading to depleted base cations and decreased BS, and further lowered ANC. However, N addition did not alter exchangeable Al(3+) , but increased cation exchange capacity (CEC). Nitrogen addition-induced increase in SOC is suggested to contribute to both higher CEC and lower pH. We further found that increased N addition greatly decreased soil solution pH at 20 cm depth, but not at 40 cm. Furthermore, there was no evidence that Al(3+) was leaching out from the deeper soils. These unique responses in tropical climate likely resulted from: exchangeable H(+) dominating changes of soil cation pool, an exhausted base cation pool, N-addition stimulating SOC production, and N saturation. Our results suggest that long-term N addition can contribute measurably to soil acidification, and that shortage of Ca and Mg should receive more attention than soil exchangeable Al in tropical forests with elevated N deposition in the future.