Advertisement
Browse Subject Areas
?

Click through the PLOS taxonomy to find articles in your field.

For more information about PLOS Subject Areas, click here.

  • Loading metrics

Open Access

Peer-reviewed

Research Article

Effect of elevated atmospheric CO2 concentration on growth and leaf litter decomposition of Quercus acutissima and Fraxinus rhynchophylla

Abstract

The atmospheric carbon dioxide (CO2) level is expected to increase substantially, which may change the global climate and carbon dynamics in ecosystems. We examined the effects of an elevated atmospheric CO2 level on the growth of Quercus acutissima and Fraxinus rhynchophylla seedlings. We investigated changes in the chemical composition of leaf litter, as well as litter decomposition. Q. acutissima and F. rhynchophylla did not show differences in dry weight between ambient CO2 and enriched CO2 treatments, but they exhibited different patterns of carbon allocation, namely, lower shoot/root ratio (S/R) and decreased specific leaf area (SLA) under CO2-enriched conditions. The elevated CO2 concentration significantly reduced the nitrogen concentration in leaf litter while increasing lignin concentrations and carbon/nitrogen (C/N) and lignin/N ratios. The microbial biomass associated with decomposing Q. acutissima leaf litter was suppressed in CO2 enrichment chambers, while that of F. rhynchophylla was not. The leaf litter of Q. acutissima from the CO2-enriched chambers, in contrast with F. rhynchophylla, contained much lower nutrient concentrations than that of the litter in the ambient air chambers. Consequently, poorer litter quality suppressed decomposition.

Citation: Cha S, Chae H-M, Lee S-H, Shim J-K (2017) Effect of elevated atmospheric CO2 concentration on growth and leaf litter decomposition of Quercus acutissima and Fraxinus rhynchophylla. PLoS ONE 12(2): e0171197. https://doi.org/10.1371/journal.pone.0171197

Editor: John J. Kelly, Loyola University Chicago, UNITED STATES

Received: May 6, 2016; Accepted: January 18, 2017; Published: February 9, 2017

Copyright: © 2017 Cha et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All relevant data are within the paper.

Funding: The authors received no specific funding for this work.

Competing interests: The authors have declared that no competing interests exist.

Introduction

In terrestrial ecosystems, carbon dioxide (CO2) is absorbed or emitted through primary production and respiration processes [1–3]. The amount of carbon (C) stored in terrestrial ecosystems is approximately 2,060 Gt, which is roughly three times that in the atmosphere (735 Gt) [1, 4, 5]. In particular, temperate forests cover only 8% of the land area globally, but they account for approximately 40% of the total terrestrial C storage. These forests are important for sequestering C from atmospheric CO2 [6–8].

Atmospheric concentrations of CO2 have increased substantially since the Industrial Revolution as a result of human activity [9]. The CO2 concentration in the atmosphere reached 397 ppm in 2014 [10]. The rising CO2 concentration may induce global warming [9, 11–13], and it has had direct impacts on the structure and function of ecosystems, including effects on tree physiology and growth [4, 14–16]. For example, plant biomass increases under elevated CO2 conditions [16, 17], as does forest productivity [1, 18], and there have been changes in the C dynamics of various ecosystems under these conditions [19, 20]. An increasing CO2 concentration has increased the total amount of C cycling and distorted the global C balance in ecosystems [1, 2, 5–9]. Elevated CO2 concentrations stimulate photosynthesis and increase net primary production, which increases the amount of C stored in trees and soil [1, 3, 14–21], notably in mid-latitude ecosystems [18].

Many studies that have focused on aboveground tree responses have shown that CO2 enrichment can lead to differences in forest ecosystem structure and function through direct effects on tree physiology and growth [4, 14–16], changes in C cycling [19, 20], and possible shifts in climate [9, 11–13]. Moreover, elevated CO2 levels can affect litter quality by altering the nutrient concentrations of plant tissue. When grown in a CO2 fumigation system, plants, such as Betula pubescens, Fraxinus excelsior, and Acer platanoides, have lower nitrogen (N) concentrations in their tissues [22–24], which results in an increase in structural and non-structural C in leaves [24–28]. These changes in plant tissues affect decomposition and mineralization processes [29, 30] by changing litter quality parameters such as the C/N and lignin to N (lignin/N) ratios [31, 32]. Consistent negative correlations with litter decomposition rates have been reported for C/N [31] and lignin/N ratios [32]. Lignin is one of the most abundant biopolymers, and it is resistant to decomposition [33, 34]. Therefore, elevated CO2 concentrations have the potential to modify litter decomposition by changing the quality and quantity of litter, which further affects the rates of biogeochemical processes in forest environments.

Recently, several long-term experiments using free-air CO2 enrichment (FACE) systems have shown the effects of a rising atmospheric CO2 concentration on terrestrial ecosystems. FACE experiments have provided novel insights into the ecological mechanisms that control the cycling and storage of C in ecosystems. However, in South Korea, FACE systems do not exist, and there is less concern about the effects of elevated CO2 concentrations on forest ecosystem function, although many studies have shown that plant growth and litter quality in forest ecosystems will change with elevated CO2 concentration. The aim of the present experiment was to determine the impact of elevated CO2 concentrations on plant growth and changes in the litter quality of two tree species, Quercus acutissima and Fraxinus rhynchophylla, which are the dominant and most widespread species in lowland temperate forests in South Korea; the acorns from Q. acutissima are traditionally used for food, while the wood from both species is used for charcoal and architectural materials [35]. Moreover, we examined the changes in the litter decomposition rate and microbial activities that were caused by CO2-mediated changes in litter qualities. To achieve these aims, we surveyed: (1) changes in growth, litter quality, and the chemical composition of leaf litter in elevated CO2 experimental chamber; (2) and changes in the decomposition rate of litter that was collected from the elevated CO2 chamber. We anticipate that the results from these experiments will provide a basis for studying the effects of elevated CO2 concentration on temperate deciduous forests in South Korea.

Materials and methods

Raising plants and litter collection

The acorns of Q. acutissima were germinated in a 25°C incubator, and we acquired 1-year-old F. rhynchophylla seedlings from the Korea National Arboretum of the Korea Forest Service. We transplanted the seedlings of both species in rectangular pots and maintained them for one growing season in an ambient CO2 (380 ppm) chamber and an enriched atmospheric CO2 (700 ppm) chamber; 700 ppm is the predicted CO2 concentration in the next century, as determined by the Intergovernmental Panel on Climate Change in 2013. The pots (0.6 m long × 0.4 m wide × 0.3 m high) were filled with artificial soil (TKS2 Instant Plus, Floragard, Oldenburg, Germany) and a vermiculite (Verminuri, GFC, Hongseong, South Korea) mixture (2:1, v/v), and each pot was fertilized once with 330 mg L−1 N as (NH4)2SO4, 220 mg L−1 P as NaH2PO4, and 400 mg L−1 K as KCl in the form of granular and incorporated into potting soil before transplanting the seedlings. We used four pots × two experimental tree species × two CO2 treatments, which resulted in a total of 16 pots. Ten Q. acutissima seedlings and six F. rhynchophylla seedlings were transplanted in each pot, which resulted in a total of 40 and 24 seedlings, respectively, for each treatment.

The seedlings in the pots were maintained in two closed-top chamber fumigation systems (2.4 m long × 1.2 m wide × 1.5 m high), one for the CO2 treatment and one for the ambient treatment. These chambers were constructed using polycarbonate (PC) sheet (Polygal Plastic Industries Ltd., Ramat Hashofet, Israel), in a greenhouse. The elevated CO2 treatment chamber was maintained at 700–750 ppm CO2 by constantly injecting a mixture of ambient air and a high concentration of CO2. The inlet ventilation air fan system forced the air into the chambers at a rate of two air changes per min. Both the chambers were ventilated in the same way, the only difference being the addition of CO2 for the CO2 treatment chamber. The system provided a stable CO2 concentration, and the temperature and humidity were close to those outside of the chambers. The CO2 concentration was monitored using an infrared CO2 analyzer (LI-840, LI-COR, Lincoln, NE, USA). The daytime air temperature in the chamber was at the most 2.4°C higher than the temperature outside (during the summer), and it was similar to that outside the chambers at night. The relative illumination of the chamber was 0.60–0.65 of that outside of the greenhouse. The relative illumination was calculated by simultaneously measuring the illumination inside the chamber and outside the greenhouse using a digital lux meter (DX-100, INS enterprise, Taipei, Taiwan) under full sunlight condition. The experiment used isolative segregation design with pseudoreplication in the two growth chambers [36]. This design is likely to represent the risks of simple segregation in an exaggerated form, and therefore, the potential for spurious effect of the treatment is much greater. Hence, we changed the position of the pots once a month between the two growth chambers to minimize the potential source of confusion caused by the experimental design [36, 37].

Plants were cultivated for 251 d from April 3, 2007, to December 10, 2007. Senescent leaves from the ambient and CO2-elevated chambers were separately collected as they fell. The collected leaves were dried at 60°C for 48 h and then stored in desiccators. The thickness of the fallen leaves from each growth pot was measured using an outside micrometer (M110-25, Mitutoyo, Kawasaki, Japan). The leaf area was measured by a scanner (Perfection 1670, Seiko Epson, Nagano, Japan) and AutoCAD (2007, Autodesk), and the leaf weight was also measured (ED423S, Sartorius, Göttingen, Germany). The leaf weight of each individual tree was calculated using the relationship between the stem weight and leaf litter weight in the pots, because it was not possible to distinguish the litter from any particular tree, and the leaf area per individual tree was determined using the average value of the area per weight of the leaves. All analyses were conducted, using an average value per tree, with each pot as a replicate. After all of the leaves had fallen, the plants were harvested and divided into aboveground and underground portions, and then they were dried completely in an oven at 60°C. Growth analysis parameters (Table 1) were calculated as follows [38].

thumbnail
Download:
Table 1. Comparison of the growth parameters of Q. acutissima and F. rhynchophylla in the ambient air (380 ppm) and elevated CO2 (700 ppm) chambers.

https://doi.org/10.1371/journal.pone.0171197.t001

Litter decomposition in microcosms

The decomposition experiment used leaf litter that was collected from the ambient air and elevated CO2 chambers, and it was conducted in a microcosms that were created within 1-L, colorless glass bottles with detachable lids, with a 9-mm diameter aperture attached to a rubber septum. The bottles were filled with 400 g of quartz sand from which organic matter was removed via repeated washing with distilled water. The quartz sand was controlled at 90% of its water-holding capacity to maintain constant humidity in the microcosm. Approximately 2 g of dried litter from the ambient and elevated CO2 chambers was enclosed in each litter bag (10 cm × 10 cm; composed of polyvinyl chloride-coated fiberglass net with a 1.2 mm2 mesh size), which were placed on the sand in each microcosm after immersion in a soil suspension solution (fresh soil:water = 1:20, weight/volume). The microcosms were incubated in a room at constant temperature (23°C) for 255 d. The litter bags were collected five times at approximately 50-d intervals during the incubation period (45, 90, 135, 195 and 255 days, respectively). The experimental design consisted of two species × two treatments × five collections × four replicates, giving a total 80 bags and bottles. The decaying litter was carefully removed from the litter bags, and then it was divided into two parts, one of which was dried at 60°C for 48 h to determine the remaining mass, while the other was used to measure the microbial biomass. The decomposition rate constant (k) was estimated following the exponential model [39] that characterizes the weight loss. where X0 = weight (g) of litter at time 0; Xt = weight (g) of litter at time t (days); k = decomposition rate constant.

Measurement of microbial respiration of decaying litter

CO2 evolution from the decaying litter was measured at 3-d intervals for the first 97 d, and then at 7–14-d intervals thereafter. The microcosms were thoroughly flushed with CO2-free air before the evolved CO2 was collected from the decaying litter. The microcosms were incubated at 23°C for 2 h with completely closed lids, and then the evolved CO2 was sampled by inserting a syringe through the rubber septum in the lids. The CO2 concentrations were determined using an infrared CO2 analyzer (modification of LI-840, LI-COR, Lincoln, NE, USA).

Microbial biomass C and N in decaying litter

Fungal (eukaryotic) and bacterial (prokaryotic) biomasses in the decaying litter were determined by the substrate-induced respiration (SIR) method, as described in Beare et al. [40], as adapted from Anderson and Domsch [41]. Chopped fresh litter samples (0.2–0.5 g) were placed into four 100-mL bottles and incubated at 4°C, followed by 2.5 mL g−1 additions of one the following solutions: (1) cycloheximide (16.0 g L−1), (2) streptomycin (3.2 g L−1), (3) cycloheximide (16.0 g L−1) + streptomycin (3.2g L−1), or (4) pure water only. Cycloheximide and streptomycin were used as eukaryotic and prokaryotic inhibitors, respectively. Additionally, 2.5 mL g−1 of a glucose solution (16.0 g L−1) was added to the bottles. The bottles were immediately sealed and incubated at 23°C for 2 h. After incubation, evolved CO2 from the litter was measured using an infrared CO2 analyzer as described above. Pure water-treated microbial biomass carbon (Cmic) was calculated using the formula of Beare et al. [40].

Bacterial and fungal contributions to the total biomass were calculated using the aforementioned streptomycin and cycloheximide treatments [40, 42].

RB is the streptomycin-treated SIR rate; RF is the cycloheximide-treated SIR, and RBF is both treated SIR rate. Fungal and bacterial biomasses were calculated according to the above equation.

Microbial biomass nitrogen (Nmic) was determined by the modified chloroform fumigation extraction method [43]. The freshly chopped litter sample was divided into two portions, one of which was extracted with 40 mL of 0.5 M K2SO4 with shaking for 1 h, followed by filtration through Whatman filter paper (no. 42), while the other subsample was fumigated with ethanol-free chloroform for 24 h in darkness, and then the chloroform was evacuated. The samples were extracted with 0.5 M K2SO4 and filtered. A 0.75-mL aliquot of the K2SO4 extract, 1.75 mL of citric acid buffer, and 1.25 mL of freshly prepared ninhydrin reagent (composed of ninhydrin, dimethyl sulfoxide, and lithium acetate buffer) were placed in a 15-mL test tube and mixed, and then boiled for 15 min in a water bath. After cooling, 5 mL of a 1:1 water and ethanol solution was added, and the optical density was determined at 570 nm. Ninhydrin-reactive N was calculated using L-leucine standards. Microbial biomass N was calculated according to Joergensen and Brookes [44].

Chemical analysis of leaf litter

The chemical composition of the litter was determined on milled litter samples. To determine the nutrient concentration, the litter materials were processed by the wet digestion method using 10 mL of HNO3 and 3 mL of HClO4 [45]. The concentrations of P, K, Ca, Mg, and Na in the digested aliquots were determined by inductively coupled plasma atomic emission spectroscopy (ICPS-7510, Shimadzu Corp., Kyoto, Japan). Lignin and cellulose concentrations were determined by the acid detergent fiber / 72% H2SO4 method [46]. Approximately 0.5 g of a milled litter sample was weighed (W1) and boiled for 1 h in a 100-mL cetyltrimethylammonium bromide solution. The solution was filtered through a pre-weighed sinter (W2) and washed three times with hot distilled water. Then, it was washed with acetone, dried for 2 h at 105°C, and weighed (W3). Approximately 10 mL of 72% H2SO4 was added to the sinter, and the mixture was kept in 72% H2SO4 for 3 h. Subsequently, the acid was removed under vacuum, and the residue was washed with hot distilled water until it was acid-free (acid-detergent fiber). The sinter was dried at 105°C for 2 h and weighed (W4). Then, the sinter was heated at 550°C for 2 h, cooled, and weighed to determine the ash content of the residue (W5). Lignin (%) and cellulose (%) were calculated as follows.

Soluble carbohydrates (CHOs) were determined by the anthrone method after hot water extraction [47]. Total C and N were determined by elemental analyzers (Flash EA 1112, Thermo Fisher Scientific, Waltham, MA, and EA1110, CE Instruments, Wigan, UK).

Data analysis

All the data analyses were performed, using average values, with each pot considered an experimental replicate. The growth parameters and chemical concentrations of leaf litter data were analyzed for statistically significant differences between the treatments, as determined by a Student's t-test. Remaining mass, microbial biomass C and N, fungal and bacterial biomass data were analyzed via repeated measures ANOVAs (rmANOVAs). Each species was analyzed individually with treatment as the within subjects factor. Greenhouse-Geisser corrections were used when the assumption of sphericity was violated. Litter quality (C, N, P, K, Ca, soluble carbohydrate, cellulose, lignin, C/N, and lignin/N) and microbial biomass parameters (microbial biomass C and N, fungal and bacterial biomass, and microbial respiration) were compared using a multivariate analysis of variance (MANOVA) with the treatment of each tree species, and a 2-way MANOVA with a species × treatment interaction (represented by Q. acutissima + F. rhynchophylla) using SPSS for Windows, version 21 (IBM Corp., Armonk, NY, USA). Additionally, we conducted a non-metric multidimensional scaling (NMDS) analysis to examine the relationships between decaying leaf litter and initial litter quality, using metaMDS and envfit in the vegan library running the R package (ver. 2.15.3, www.r-project.org). For all analyses, differences among groups were considered significant if p < 0.05.

Results

Plant growth in the elevated CO2 chamber

The growth of Q. acutissima and F. rhynchophylla did not statistically differ between the ambient air and elevated CO2 chambers (Table 1). However, the shoot/root (S/R) ratio differed between the two species. The S/R ratio of Q. acutissima was significantly lower in elevated CO2 chamber. Leaf growth conspicuously differed between the conditions. The thickness of the leaf blade was significantly higher (22% for Q. acutissima and 15% for F. rhynchophylla) in the elevated CO2 chamber. The leaf area of F. rhynchophylla litter was significantly lower in elevated CO2 chamber. And the specific leaf area (SLA) of the leaf litter of Q. acutissima and F. rhynchophylla were significantly lower at the higher CO2 concentration (Table 1).

Changes in the chemical composition of leaf litter

The C concentration of the leaf litter of Q. acutissima that was grown in the ambient and elevated CO2 chambers did not differ significantly (Table 2). However, the N concentration of the leaf litter was significantly lower in the elevated CO2 chamber. Conversely, in the F. rhynchophylla litter, the C concentration was higher in the elevated CO2 chamber, while the N concentration did not differ significantly between the two conditions. The C/N ratio of the Q. acutissima litter increased by 1.50-fold in the elevated CO2 chamber, and that of F. rhynchophylla litter did not significant but showed an increasing trend. The P concentration of the leaf litter differed between the two tree species. The P of the Q. acutissima litter was lower in the elevated CO2 chamber, while that of F. rhynchophylla was higher. However, the calcium concentration of the leaf litter was significantly lower in the elevated CO2 chamber for both species (Table 2). The cellulose concentration in the leaf litter of Q. acutissima was significantly lower under high CO2, while the soluble CHO and lignin concentrations were significantly higher in the elevated CO2 chamber (1.42-fold and 1.20-fold for Q. acutissima, 1.12-fold and 1.13-fold F. rhynchophylla, respectively). The lignin/N ratio of the Q. acutissima leaf litter was remarkably higher at the higher CO2 concentration and F. rhynchophylla leaf litter did not significant but showed an increasing tendency as C/N ratio.

thumbnail
Download:
Table 2. Chemical concentrations of leaf litter collected from the ambient air and elevated CO2 chambers.

https://doi.org/10.1371/journal.pone.0171197.t002

Litter decomposition

The mass losses of the two species’ leaf litter following incubation at 23°C for 255 d in the microcosms are shown in Fig 1. The leaf litter from the elevated CO2 chamber exhibited a significantly lower decay rate than the litter from the ambient air chamber. The mass losses of the Q. acutissima and F. rhynchophylla litter in the ambient air chamber were 34.6% and 40.9%, respectively, and the Q. acutissima and F. rhynchophylla litter from the elevated CO2 chamber decomposed by 27.8% and 34.6%, respectively (Fig 1). The decomposition rate constants (k) of the leaf litter from the elevated CO2 chamber were significantly smaller than that in the ambient air chamber. The k values of the Q. acutissima and F. rhynchophylla litter in the ambient air chamber were 0.0014 yr−1 and 0.0021 yr−1, respectively, and those of the litter from the elevated CO2 chamber were 0.001 yr−1 and 0.0016 yr−1, respectively.

thumbnail
Download:
Fig 1. Changes in the remaining mass of the Q. acutissima (upper) and F. rhynchophylla (lower) leaf litter in the microcosms.

Closed circles represent leaf litter obtained from the ambient air (380 ppm CO2) chamber, and open circles represent that collected from the elevated CO2 (700 ppm) chamber. Bars show standard deviations (n = 4).

https://doi.org/10.1371/journal.pone.0171197.g001

Microbial biomass and activity of decaying litter

Changes in the microbial biomass C and N during the decomposition of the leaf litter of both species, which was collected from the ambient air and elevated CO2 chambers in the laboratory microcosm, are summarized in Fig 2. The microbial biomass C in the leaf litter of Q. acutissima was significantly greater in the litter from the ambient air chamber, compared with that from the elevated CO2 chamber, while the microbial biomass C in the leaf litter of F. rhynchophylla was significantly lower than elevated CO2 chamber. However, microbial biomass N in the leaf litter of both species did not differ significantly between the elevated CO2 and ambient air chambers.

thumbnail
Download:
Fig 2. Changes in the microbial biomass C and N of the decomposing leaf litter in the microcosms.

Closed circles show leaf litter obtained from the ambient air (380 ppm CO2) chamber, and open circles represent leaf litter obtained from the elevated CO2 (700 ppm) chamber. Bars show standard deviations (n = 4).

https://doi.org/10.1371/journal.pone.0171197.g002

There were differences in the microbial biomass change during the decomposition of the two leaf litter species. The microbial biomass C and N in the decaying Q. acutissima leaf litter were nearly stable throughout the experimental period of 255 d. However, during the decomposition of F. rhynchophylla leaf litter, the microbial biomass N increased gradually throughout the experimental period.

Changes in the fungal and bacterial biomasses in the decaying litter differed according to litter species and treatment (Fig 3). The bacterial biomass showed a tendency to peak 90 d into the experiment, whereas the fungal biomass peaked at the end of the experiment. In the Q. acutissima leaf litter, the fungal and bacterial biomasses were greater in the litter from the ambient air chamber, compared with that from the elevated CO2 chamber. However, the fungal biomass of the F. rhynchophylla leaf litter was lower in the elevated CO2 chamber. And bacterial biomasses of the F. rhynchophylla leaf litter occasionally varied between the ambient air and elevated CO2 chambers. These results suggest that there was a succession of microbial groups during the course of the experiment; the bacterial group increased during the early decay stages, while the fungal group dominated after the early stages of leaf litter decomposition for both tree species.

thumbnail
Download:
Fig 3. Changes in the fungal and bacterial biomasses in the decomposing leaf litter in the microcosms.

Closed circles represent leaf litter obtained from the ambient air (380 ppm CO2) chamber, and open circles represent leaf litter obtained from the elevated CO2 (700 ppm) chamber. Bars show standard deviations (n = 4).

https://doi.org/10.1371/journal.pone.0171197.g003

Microbial respiration in the decomposing leaf litter of the two species was active for 1 month, peaking 15 d after incubation. However, the amount of respiration was about two times greater in the Q. acutissima leaf litter than in the F. rhynchophylla litter, and it was significantly lower in the litter from the elevated CO2 chamber than in the ambient air chamber. Differences of microbial respiration between the two types of litter were greater at approximately 15 d of incubation, and small differences appeared gradually as litter decay progressed (Fig 4). These differences in microbial respiration between the litter from the ambient air and elevated CO2 chambers coincided more with the litter mass loss trends than with the parameters of microbial biomass.

thumbnail
Download:
Fig 4. Changes in CO2 evolution from the decomposing leaf litter in the microcosms.

Closed circles represent leaf litter obtained from the ambient air (380 ppm CO2) chamber, and open circles represent leaf litter obtained from the elevated CO2 (700 ppm) chamber. Bars show standard deviations (n = 4).

https://doi.org/10.1371/journal.pone.0171197.g004

Factors affecting leaf litter decomposition

Litter quality (C, N, P, K, Ca, soluble CHO, cellulose, lignin, C/N, and lignin/N) was significantly affected by the elevated CO2 (Table 3). And the microbial biomass parameters of decaying Q. acutissima and F. rhynchophylla leaf litter showed also significant differences between the two treatments (Table 3).

thumbnail
Download:
Table 3. Summary of the MANOVA results for the effect of elevated CO2 on Q. acutissima and F. rhynchophylla.

https://doi.org/10.1371/journal.pone.0171197.t003

The results of the NMDS, which identified the relative effects of each parameter on litter decomposition, are shown in Fig 5. In the NMDS analysis, the litter from each CO2 treatment of Q. acutissima and F. rhynchophylla formed a distinct cluster that was related to mass loss (Fig 5). The vectors of the litter quality factors were fitted to the extent of the differences of the litter decomposition rate in ordination space. The C, N, and lignin concentrations were correlated with the ordination (r2 = 0.524, 0.722, and 0.902, respectively; p < 0.001), with arrows pointing in the direction of the Q. acutissima group. Moreover, the cellulose and soluble CHO concentrations were significantly correlated with mass loss (r2 = 0.802 and 0.546, respectively; p < 0.001), with arrows pointing toward the F. rhynchophylla group. For the C/N, lignin/N ratios, and microbial respiration, the arrows also pointed in the direction of the F. rhynchophylla group (r2 = 0.841, 0.902, and 0.524, respectively; p < 0.01). Microbial biomass C and N, fungal and bacterial biomasses did not significantly correlate with mass loss.

thumbnail
Download:
Fig 5. Ordination biplot of NMDS based on the leaf litter decomposition rate of each species.

Angles and lengths of the radiating arrows indicate the direction and strength of the relationships of the litter quality parameters. Each vector's significance level was less than 0.05. Carbon (C), nitrogen (N), lignin (Lig), cellulose (Cel), soluble carbohydrate (SC), C/N (CN), lignin/N (LigN), respiration (RES), cQa (ambient CO2 Q. acutissima group), eQa (elevated CO2 Q. acutissima group), cFr (ambient CO2 F. rhynchophylla group), and eFr (elevated CO2 F. rhynchophylla group).

https://doi.org/10.1371/journal.pone.0171197.g005

Discussion

The objective of the present study was to investigate the effect of elevated CO2 concentration on the growth and litter quality of Q. acutissima and F. rhynchophylla, and to determine the changes in litter decomposition according to any CO2-mediated change in the litter quality. Our results did not show a statistically significant difference in plant growth, as evidenced by the total dry weight, between the ambient and elevated CO2 concentrations. The S/R ratio of the tree species was lower under elevated CO2. Changes in plant growth response to high CO2 concentrations depend on the growth conditions or growth potential of a species [16, 21, 48, 49], as well as the species’ developmental strategy, such as the creation of new sinks for extra C [50]. Several studies have suggested that growth under elevated CO2 concentrations leads to shifts in the root system architecture, which could enhance the nutrient uptake capacity [51–53] by increasing the biomass and density of the fine roots [3, 54–56], as well as the S/R ratio [50]. Our results are consistent with the aforementioned studies. In our experiments, the S/R ratio for Q. acutissima was clearly lower at the elevated CO2 concentration. It is evident that an elevated CO2 concentration can reduce the stem proportion and increase the root proportion, at least in young tree seedlings in some species, without changing the mass.

High atmospheric CO2 concentrations affect the physical structure of leaf litter [3, 57, 58]. In the present study, for both Q. acutissima and F. rhynchophylla, the leaf thickness was higher and the leaf area was lower under the elevated CO2 concentration, without changes in leaf mass. This is consistent with the results of many studies [59–63]. However, Pritchard et al. [64] showed variable differences according to species, highlighting that leaf thickness typically increased (81%) or was occasionally unaffected (19%) by increased CO2 concentrations.

Litter quality, such as the concentrations of nutrients and their composition, of the young Q. acutissima and F. rhynchophylla seedlings was also significantly affected by the elevated CO2 concentration (Table 3). The elevated CO2 treatment of the two tree seedlings altered the chemical composition of the leaf litter, e.g., it decreased the N concentration and increased the C/N ratio in Q. acutissima, and increased the lignin and soluble CHO concentrations in Q. acutissima and F. rhynchophylla. Many studies have also asserted that the N concentration of a plant tissue is reduced and the C/N ratio is increased by CO2 enrichment [65, 66], owing to a physiological response that increases the efficiency of N use in plants. This leads to a greater production of biomass per mole of N taken up [66, 67]. Moreover, CO2 enrichment generally increases the concentration of non-structural C, such as soluble CHOs [68] and secondary compounds [69, 70], through enhanced production of photosynthetic products and biomass dilution [71]. Elevated CO2 concentrations also increase the lignin concentration [24, 25, 28, 72–75], while decreasing the cellulose concentration in leaf litter. This, in turn, increases the lignin/N ratio significantly, which is used as a predictive index of decomposition [31, 32, 34, 72, 76]. Therefore, a decreased N concentration and increased lignin concentration and lignin/N ratio, which are induced by an increase in the atmospheric CO2 concentration, could reduce the litter decomposition rate [30, 34, 77, 78] by preventing an increase in microbial biomass, as well as microbial activities, by changing the litter quality [34, 79, 80].

During litter decomposition, both the chemical composition of the litter (its quality) and microbial biomass and its activity are very important factors [81, 82]. The leaf litter decomposition of the two tree species was significantly affected by changes in the litter quality that were induced by the CO2 treatment (Table 3). These changes in litter quality, such as the N and lignin concentrations, the C/N ratio, and the lignin/N ratio are considered to be major factors affecting litter decomposition [30–32, 34, 79]. A decrease in the N concentration and an increase in the lignin concentration change the quality of leaf litter, which affect the decomposer activities of microorganisms [83]. The microbial biomass in the litter was altered due to elevated CO2 treatments (Table 3), but there was not significantly affect the litter decomposition rate (Fig 5). It is presumably the result of a species-specific responses on decomposition [24]. Microbial succession [34, 77–80, 84], i.e., alteration of bacterial and fungal biomasses, differed significantly between the ambient and elevated CO2 treatments, but the microbial biomass and activity (respiration), which were altered by the elevated CO2 treatment, had weaker effects on litter decomposition than the litter quality.

In present study, although with potential error due to pseudoreplication, the litter produced under elevated CO2 conditions showed a tendency to decompose more slowly due to CO2-induced changes in the litter quality. The decreasing litter decomposition rate may expedite C deposition on the forest floor. Long-term experiments under FACE have produced findings related to the C cycle. Norby and Zak [85] reviewed the effect of CO2 on C cycling, and they suggested that an elevated CO2 concentration does not necessarily increase ecosystem C storage. However, there are many indications that C accumulates in either plant biomass or soil organic matter in certain ecosystems or under certain conditions [86–90]. By examining the effect of elevated CO2 conditions, compared with ambient CO2 conditions, Luo et al. [88] showed that the average C pools in shoots, roots, and whole plants increased by 22.4, 31.6, and 23.0%, respectively. Similarly, via a meta-analysis, they found that litter and soil C pools increased by 20.6 and 5.6% under elevated CO2 concentrations. Generally, forest ecosystems store a large amount of C in the form of humus. The humus can constitute as much 30 percent of the organic matter in some soil [91]. Many researchers have suggested that such an accumulation would continue for millennia in the absence of a disturbance, which would result in the buildup of considerable amounts of humus [84]. Additional reductions in decomposition rates of two species from elevated CO2, resulting from increased anthropogenic CO2 emissions, may have an impact on increase C accumulation in temperate deciduous forest in South Korea.

Based on the above results and our experimental results, the effects of increasing CO2 concentrations on plant growth vary by tree species (Q. acutissima and F. rhynchophylla). Nevertheless, by elevated CO2, more photosynthetic products were allocated to roots (which decreased the S/R ratio), and leaves became thicker and smaller at the elevated CO2 concentrations. Furthermore, litter quality was altered by the elevated CO2 concentration because of the lower N concentration or the higher lignin concentration in the leaf litter. Litter from the elevated CO2 treatment decayed more slowly; thus, we believe that C deposition in the soil accelerated under these conditions. In our study, we only investigated the direct effects of an elevated CO2 concentration. However, in addition to increasing CO2 concentrations, other environmental factors (climate and atmospheric environment) can affect plants [9], and these changes also impact ecosystems [9, 92–94]. Changes in factors such as temperature and precipitation have a major impact on the distribution and activity of organisms, which is a major concern of many researchers [95–101]. Recently, several studies have examined the combined effects of elevated atmospheric CO2 concentrations and other environmental factors, especially temperature [16, 98, 102, 103], light [104, 105], and water [106]. Nevertheless, studies of such combined effects are scarce. The combined effects of environmental factors and elevated CO2 concentrations on forest ecosystem are uncertain because they are very complex, especially regarding C cycling [107, 108]. Therefore, additional research is needed to understand the effects of other factors in combination with elevated CO2 concentrations.

Author Contributions

  1. Conceptualization: JKS SC HMC.
  2. Data curation: JKS SC.
  3. Formal analysis: SC.
  4. Funding acquisition: JKS SC.
  5. Investigation: SC.
  6. Methodology: SC HMC.
  7. Project administration: SC.
  8. Resources: SC HMC.
  9. Software: SC SHL.
  10. Supervision: JKS SC.
  11. Validation: JKS SC HMC.
  12. Visualization: SC.
  13. Writing – original draft: SC.
  14. Writing – review & editing: JKS SC.

References

  1. 1. Schimel DS. Terrestrial ecosystems and the carbon-cycle. Glob Chang Biol. 1995;1(1):77–91.
  2. 2. Keeling CD, Chin JFS, Whorf TP. Increased activity of northern vegetation inferred from atmospheric CO2 measurements. Nature. 1996;382(6587):146–9.
  3. 3. Reverchon F, Xu Z, Blumfield TJ, Chen C, Abdullah KM. Impact of global climate change and fire on the occurrence and function of understorey legumes in forest ecosystems. J Soils Sediments. 2012;12(2):150–60.
  4. 4. Ceulemans R. Effects of CO2 enrichment on trees and forests: Lessons to be learned in view of future ecosystem studies. Ann Bot. 1999;84(5):577–90.
  5. 5. Hu S, Chapin FS 3rd, Firestone MK, Field CB, Chiariello NR. Nitrogen limitation of microbial decomposition in a grassland under elevated CO2. Nature. 2001;409(6817):188–91. pmid:11196641
  6. 6. Martin PH, Nabuurs G-J, Aubinet M, Karjalainen T, Vine EL, Kinsman J, et al. Carbon sinks in temperate forests. Annu Rev Ecol Evol Syst. 2001;26(1):435–65.
  7. 7. Wamelink GWW, Wieggers HJJ, Reinds GJ, Kros J, Mol-Dijkstra JP, van Oijen M, et al. Modelling impacts of changes in carbon dioxide concentration, climate and nitrogen deposition on carbon sequestration by European forests and forest soils. For Ecol Manage. 2009;258(8):1794–805.
  8. 8. Tyrrell ML, Ross J, Kelty M. Carbon dynamics in the temperate forest. Managing forest carbon in a changing climate: Springer; 2012.
    • 9. Intergovernmental Panel on Climate Change. Climate change 2013: the physical science basis: Working Group I contribution to the Fifth assessment report of the Intergovernmental Panel on Climate Change. Cambridge: Cambridge University Press; 2014.
      • 10. WMO Greenhouse Gas Bulletin. The state of greenhouse gases in the atmosphere using global observations through 2014. 2015; Available: www.wmo.int/pages/prog/arep/gaw/ghg/GHGbulletin.html.
        • 11. Heimann M, Reichstein M. Terrestrial ecosystem carbon dynamics and climate feedbacks. Nature. 2008;451(7176):289–92. pmid:18202646
        • 12. Allen MR, Frame DJ, Huntingford C, Jones CD, Lowe JA, Meinshausen M, et al. Warming caused by cumulative carbon emissions towards the trillionth tonne. Nature. 2009;458(7242):1163–6. pmid:19407800
        • 13. Meinshausen M, Meinshausen N, Hare W, Raper SC, Frieler K, Knutti R, et al. Greenhouse-gas emission targets for limiting global warming to 2 degrees C. Nature. 2009;458(7242):1158–62. pmid:19407799
        • 14. Bazzaz FA. The response of natural ecosystems to the rising global CO2 levels. Annu Rev Ecol Evol Syst. 1990;21:167–96.
        • 15. Jarvis PG, Aitken AM. European forests and global change: the likely impacts of rising CO2 and temperature: Cambridge University Press; 1998.
          • 16. Lopes A, Ferreira AB, Pantoja PO, Parolin P, Piedade MTF. Combined effect of elevated CO2 level and temperature on germination and initial growth of Montrichardia arborescens (L.) Schott (Araceae): a microcosm experiment. Hydrobiologia. 2015:1–12.
          • 17. Tissue DT, Thomas RB, Strain BR. Atmospheric CO2 enrichment increases growth and photosynthesis of Pinus taeda: a 4 year experiment in the field. Plant Cell Environ. 1997;20(9):1123–34.
          • 18. Fan S, Gloor M, Mahlman J, Pacala S, Sarmiento J, Takahashi T, et al. A large terrestrial carbon sink in north america implied by atmospheric and oceanic carbon dioxide data and models. Science. 1998;282(5388):442–6. pmid:9774264
          • 19. Körner C, Asshoff R, Bignucolo O, Hattenschwiler S, Keel SG, Pelaez-Riedl S, et al. Carbon flux and growth in mature deciduous forest trees exposed to elevated CO2. Science. 2005;309(5739):1360–2. pmid:16123297
          • 20. Loudermilk EL, Scheller RM, Weisberg PJ, Yang J, Dilts TE, Karam SL, et al. Carbon dynamics in the future forest: the importance of long-term successional legacy and climate–fire interactions. Glob Chang Biol. 2013;19(11):3502–15. pmid:23821586
          • 21. de la Mata L, Cabello P, de la Haba P, Agüera E. Growth under elevated atmospheric CO2 concentration accelerates leaf senescence in sunflower (Helianthus annuus L.) plants. J Plant Physiol. 2012;169(14):1392–400. pmid:22818664
          • 22. Cotrufo MF, Briones MaJI, Ineson P. Elevated CO2 affects field decomposition rate and palatability of tree leaf litter: Importance of changes in substrate quality. Soil Biol Biochem. 1998;30(12):1565–71.
          • 23. Coûteaux M-M, Mousseau M, Célérier M-L, Bottner P. Increased atmospheric CO2 and litter quality: Decomposition of sweet chestnut leaf litter with animal food webs of different complexities. Oikos. 1991;61(1):54–64.
          • 24. Cotrufo MF, Ineson P, Rowland AP. Decomposition of tree leaf litters grown under elevated CO2—Effect of litter quality. Plant Soil. 1994;163(1):121–30.
          • 25. Cotrufo MF, Ineson P. Elevated CO2 reduces field decomposition rates of Betula pendula (Roth) leaf litter. Oecologia. 1996;106(4):525–30.
          • 26. Heagle AS, Burns JC, Fisher DS, Miller JE. Effects of carbon dioxide enrichment on leaf chemistry and reproduction by two spotted spider mites (Acari: Tetranychidae) on white clover. Environ Entomol. 2002;31(4):594–601.
          • 27. Körner C. Carbon limitation in trees. J Ecol. 2003;91(1):4–17.
          • 28. Kasurinen A, Riikonen J, Oksanen E, Vapaavuori E, Holopainen T. Chemical composition and decomposition of silver birch leaf litter produced under elevated CO2 and O3. Plant Soil. 2006;282(1):261–80.
          • 29. de Angelis P, Chigwerewe KS, Mugnozza GES. Litter quality and decomposition in a CO2-enriched Mediterranean forest ecosystem. Plant Soil. 2000;224(1):31–41.
          • 30. Cornwell WK, Cornelissen JHC, Amatangelo K, Dorrepaal E, Eviner VT, Godoy O, et al. Plant species traits are the predominant control on litter decomposition rates within biomes worldwide. Ecol Lett. 2008;11(10):1065–71. pmid:18627410
          • 31. Taylor BR, Parkinson D, Parsons WFJ. Nitrogen and lignin content as predictors of litter decay rates—a microcosm test. Ecology. 1989;70(1):97–104.
          • 32. Melillo JM, Aber JD, Muratore JF. Nitrogen and lignin control of hardwood leaf litter decomposition dynamics. Ecology. 1982;63(3):621–6.
          • 33. Meentemeyer V. Macroclimate the lignin control of litter decomposition rates. Ecology. 1978;59(3):465–72.
          • 34. Talbot JM, Treseder KK. Interactions among lignin, cellulose, and nitrogen drive litter chemistry–decay relationships. Ecology. 2012;93(2):345–54. pmid:22624316
          • 35. Lee T. Coloured Flora of Korea, Vol I, II. Seoul: Hayangmunsa; 2003.
            • 36. Hurlbert SH. Pseudoreplication and the Design of Ecological Field Experiments. Ecol Monogr. 1984;54(2):187–211.
            • 37. Pardos M, Puértolas J, Aranda I, Pardos JA. Can CO2 enrichment modify the effect of water and high light stress on biomass allocation and relative growth rate of cork oak seedlings? Trees. 2006;20(6):713–24.
            • 38. Hunt R. Basic plant growth analysis. Unwin, Hyman, London; 1990.
              • 39. Olson JS. Energy storage and the balance of producers and decomposers in ecological systems. Ecology. 1963;44(2):322–331.
              • 40. Beare MH, Neely CL, Coleman DC, Hargrove WL. Characterization of a substrate-induced respiration method for measuring fungal, bacterial and total microbial biomass on plant residues. Agr Ecosyst Environ. 1991;34(1–4):65–73.
              • 41. Anderson JPE, Domsch KH. A physiological method for the quantitative measurement of microbial biomass in soils. Soil Biol Biochem. 1978;10(3):215–21.
              • 42. Nakamoto T, Wakahara S. Development of Substrate Induced Respiration (SIR) Method Combined with Selective Inhibition for Estimating Fungal andBacterial Biomass in Humic Andosols. Plant Prod Sci. 2004;7(1):70–6.
              • 43. Brookes PC, Landman A, Pruden G, Jenkinson DS. Chloroform fumigation and the release of soil-nitrogen—a rapid direct extraction method to measure microbial biomass nitrogen in soil. Soil Biol Biochem. 1985;17(6):837–42.
              • 44. Joergensen RG, Brookes PC. Ninhydrin-reactive nitrogen measurements of microbial biomass in 0.5-M K2SO4 soil extracts. Soil Biol Biochem. 1990;22(8):1023–7.
              • 45. Helrich K. Official methods of Analysis of the AOAC. Volume 1: Association of Official Analytical Chemists Inc.; 1990.
                • 46. Rowland AP, Roberts JD. Lignin and cellulose fractionation in decomposition studies using acid‐detergent fibre methods. Commun Soil Sci Plant Anal. 1994;25(3–4):269–77.
                • 47. Allen SE, Grimshaw H, Parkinson JA, Quarmby C. Chemical analysis of ecological materials. Oxford: Blackwell Scientific; 1974.
                  • 48. Eamus D, Jarvis PG. The direct effects of increase in the global atmospheric CO2 concentration on natural and commercial temperate trees and forests. Adv Ecol Res. Volume 34: Academic Press; 2004.
                  • 49. Reich PB, Hobbie SE, Lee TD. Plant growth enhancement by elevated CO2 eliminated by joint water and nitrogen limitation. Nature Geosci. 2014;7(12):920–4.
                  • 50. Ceulemans R, Mousseau M. Tansley Review No. 71 Effects of elevated atmospheric CO2 on woody plants. New Phytol. 1994;127(3):425–46.
                  • 51. Lynch JP, St Clair SB. Mineral stress: the missing link in understanding how global climate change will affect plants in real world soils. Field Crops Res. 2004;90(1):101–15.
                  • 52. Li T, Di Z, Han X, Yang X. Elevated CO2 improves root growth and cadmium accumulation in the hyperaccumulator Sedum alfredii. Plant Soil. 2012;354(1):325–34.
                  • 53. Bloom AJ, Burger M, Kimball BA, Pinter PJ Jr.. Nitrate assimilation is inhibited by elevated CO2 in field-grown wheat. Nature Clim Change. 2014;4(6):477–80.
                  • 54. Pendall E, Bridgham S, Hanson PJ, Hungate B, Kicklighter DW, Johnson DW, et al. Below-ground process responses to elevated CO2 and temperature: a discussion of observations, measurement methods, and models. New Phytol. 2004;162(2):311–22.
                  • 55. Madhu M, Hatfield JL. Dynamics of plant root growth under increased atmospheric carbon dioxide. Agron J. 2013;105(3):657–69.
                  • 56. Smith AR, Lukac M, Bambrick M, Miglietta F, Godbold DL. Tree species diversity interacts with elevated CO2 to induce a greater root system response. Glob Chang Biol. 2013;19(1):217–28. pmid:23504733
                  • 57. Cotrufo MF, Berg B, Kratz W. Increased atmospheric CO2 and litter quality. Environmental Reviews. 1998;6(1):1–12.
                  • 58. Norby RJ, Wullschleger SD, Gunderson CA, Johnson DW, Ceulemans R. Tree responses to rising CO2 in field experiments: implications for the future forest. Plant Cell Environ. 1999;22(6):683–714.
                  • 59. Faria T, Wilkins D, Besford RT, Vaz M, Pereira JS, Chaves MM. Growth at elevated CO2 leads to down-regulation of photosynthesis and altered response to high temperature in Quercus suber L seedlings. J Exp Bot. 1996;47(304):1755–61.
                  • 60. Teng N, Wang J, Chen T, Wu X, Wang Y, Lin J. Elevated CO2 induces physiological, biochemical and structural changes in leaves of Arabidopsis thaliana. New Phytol. 2006;172(1):92–103. pmid:16945092
                  • 61. Xu CY, Salih A, Ghannoum O, Tissue DT. Leaf structural characteristics are less important than leaf chemical properties in determining the response of leaf mass per area and photosynthesis of Eucalyptus saligna to industrial-age changes in [CO2] and temperature. J Exp Bot. 2012.
                  • 62. Murray TJ, Ellsworth DS, Tissue DT, Riegler M. Interactive direct and plant-mediated effects of elevated atmospheric [CO2] and temperature on a eucalypt-feeding insect herbivore. Glob Chang Biol. 2013;19(5):1407–16. pmid:23504696
                  • 63. Jiang S, Liu T, Yu F, Li T, Parajulee MN, Zhang L, et al. Feeding behavioral response of cotton aphid, Aphis gossypii, to elevated CO2: EPG test with leaf microstructure and leaf chemistry. Entomol Exp et Appl. 2016:n/a-n/a.
                  • 64. Pritchard SG, Rogers HH, Prior SA, Peterson CM. Elevated CO2 and plant structure: a review. Glob Chang Biol. 1999;5(7):807–37.
                  • 65. Larsen KS, Andresen LC, Beier C, Jonasson S, Albert KR, Ambus P, et al. Reduced N cycling in response to elevated CO2, warming, and drought in a Danish heathland: Synthesizing results of the CLIMAITE project after two years of treatments. Glob Chang Biol. 2011;17(5):1884–99.
                  • 66. Yuan ZY, Chen HYH. Decoupling of nitrogen and phosphorus in terrestrial plants associated with global changes. Nature Clim Change. 2015;5(5):465–9.
                  • 67. Kohen AE, Rouhier H, Mousseau M, editors. Changes in dry weight and nitrogen partitioning induced by elevated CO2 depend on soil nutrient availability in sweet chestnut (Castanea sativa Mill). Annales des sciences forestières; 1992: EDP Sciences.
                    • 68. Sharma N, Sinha PG, Bhatnagar AK. Effect of elevated [CO2] on cell structure and function in seed plants. Climate Change and Environmental Sustainability. 2014;2(2):69–104.
                    • 69. Entry JA, Runion GB, Prior SA, Mitchell RJ, Rogers HH. Influence of CO2 enrichment and nitrogen fertilization on tissue chemistry and carbon allocation in longleaf pine seedlings. Root demographics and their efficiencies in sustainable agriculture, grasslands and forest ecosystems: Springer; 1998.
                      • 70. Gebauer ERL, Strain RB, Reynolds FJ. The effect of elevated CO2 and N availability on tissue concentrations and whole plant pools of carbon-based secondary compounds in loblolly pine (Pinus taeda). Oecologia. 1997;113(1):29–36.
                      • 71. Taub DR, Wang X. Why are nitrogen concentrations in plant tissues lower under elevated CO2? A critical examination of the hypotheses. J Integr Plant Biol. 2008;50(11):1365–74. pmid:19017124
                      • 72. Melillo J. Will increases in atmospheric CO2 concentrations affect decay processes. Ecosystems Center Annual Report. 1983:10–1.
                        • 73. Lee TD, Reich PB, Tjoelker MG. Legume presence increases photosynthesis and N concentrations of co-occurring non-fixers but does not modulate their responsiveness to carbon dioxide enrichment. Oecologia. 2003;137(1):22–31. pmid:12802677
                        • 74. Ziska LH, Reeves JB, Blank B. The impact of recent increases in atmospheric CO2 on biomass production and vegetative retention of Cheatgrass (Bromus tectorum): implications for fire disturbance. Glob Chang Biol. 2005;11(8):1325–32.
                        • 75. Richet N, Afif D, Tozo K, Pollet B, Maillard P, Huber F, et al. Elevated CO2 and/or ozone modify lignification in the wood of poplars (Populus tremula x alba). J Exp Bot. 2012;63(11):4291–301. pmid:22553285
                        • 76. Norby RJ, Cotrufo MF, Ineson P, O'Neill EG, Canadell JG. Elevated CO2, litter chemistry, and decomposition: a synthesis. Oecologia. 2001;127(2):153–65. pmid:24577644
                        • 77. Brandstätter C, Keiblinger K, Wanek W, Zechmeister-Boltenstern S. A closeup study of early beech litter decomposition: potential drivers and microbial interactions on a changing substrate. Plant Soil. 2013;371(1):139–54.
                        • 78. Khalsa SDS, Almanza CA, Brown PH, Smart DR. Leaf litter C and N cycling from a deciduous permanent crop. Soil Science and Plant Nutrition. 2016;62(3):271–6.
                        • 79. Zhu J, Yang W, He X. Temporal Dynamics of Abiotic and Biotic Factors on Leaf Litter of Three Plant Species in Relation to Decomposition Rate along a Subalpine Elevation Gradient. PLoS ONE. 2013;8(4):e62073. pmid:23620803
                        • 80. Wymore AS, Liu CM, Hungate BA, Schwartz E, Price LB, Whitham TG, et al. The Influence of Time and Plant Species on the Composition of the Decomposing Bacterial Community in a Stream Ecosystem. Microb Ecol. 2016;71(4):825–34. pmid:26879940
                        • 81. Swift MJ, Heal OW, Anderson JM. Decomposition in terrestrial ecosystems: Univ of California Press; 1979.
                          • 82. Parmelee RW, Beare MH, Blair JM. Decomposition and nitrogen dynamics of surface weed residues in no-tillage agroecosystems under drought conditions—Influence of resource quality on the decomposer community. Soil Biol Biochem. 1989;21(1):97–103.
                          • 83. Keiser AD, Knoepp JD, Bradford MA. Microbial communities may modify how litter quality affects potential decomposition rates as tree species migrate. Plant Soil. 2013;372(1):167–76.
                          • 84. Berg B, McClaugherty C. Plant litter. Berlin: Springer; 2008.
                            • 85. Norby RJ, Zak DR. Ecological lessons from free-air CO2 enrichment (FACE) experiments. Annu Rev Ecol Evol Syst. 2011;42(1):181–203.
                            • 86. Jastrow JD, Michael Miller R, Matamala R, Norby RJ, Boutton TW, Rice CW, et al. Elevated atmospheric carbon dioxide increases soil carbon. Glob Chang Biol. 2005;11(12):2057–64.
                            • 87. de Graaff MA, van Groenigen KJ, Six J, Hungate B, van Kessel C. Interactions between plant growth and soil nutrient cycling under elevated CO2: a meta-analysis. Glob Chang Biol. 2006;12(11):2077–91.
                            • 88. Luo Y, Hui D, Zhang D. Elevated CO2 stimulates net accumulations of carbon and nitrogen in land ecosystems: a meta-analysis. Ecology. 2006;87(1):53–63. pmid:16634296
                            • 89. van Groenigen KJ, Six J, Hungate BA, de Graaff MA, van Breemen N, van Kessel C. Element interactions limit soil carbon storage. Proceedings of the National Academy of Sciences. 2006;103(17):6571–4.
                            • 90. Sulman BN, Phillips RP, Oishi AC, Shevliakova E, Pacala SW. Microbe-driven turnover offsets mineral-mediated storage of soil carbon under elevated CO2. Nature Clim Change. 2014;4(12):1099–102.
                            • 91. Stevenson FJ. Humus chemistry: genesis, composition reactions.Hoboken, NJ: John Wiley & Sons; 1994.
                              • 92. Finzi AC, Moore DJ, DeLucia EH, Lichter J, Hofmockel KS, Jackson RB et al. Progressive nitrogen limitation of ecosystem processes under elevated CO2 in a warm-temperate forest. Ecology. 2006;87(1):15–25. pmid:16634293
                              • 93. Allen CD, Macalady AK, Chenchouni H, Bachelet D, McDowell N, Vennetier M, et al. A global overview of drought and heat-induced tree mortality reveals emerging climate change risks for forests. For Ecol Manage. 2010;259(4):660–84.
                              • 94. Kardol P, Reynolds WN, Norby RJ, Classen AT. Climate change effects on soil microarthropod abundance and community structure. Appl Soil Ecol. 2011;47(1):37–44.
                              • 95. Davidson EA, Janssens IA. Temperature sensitivity of soil carbon decomposition and feedbacks to climate change. Nature. 2006;440(7081):165–73. pmid:16525463
                              • 96. Brandt LA, King JY, Milchunas DG. Effects of ultraviolet radiation on litter decomposition depend on precipitation and litter chemistry in a shortgrass steppe ecosystem. Glob Chang Biol. 2007;13(10):2193–205.
                              • 97. Gavazov KS. Dynamics of alpine plant litter decomposition in a changing climate. Plant Soil. 2010;337(1):19–32.
                              • 98. Conant RT, Ryan MG, Ågren GI, Birge HE, Davidson EA, Eliasson PE, et al. Temperature and soil organic matter decomposition rates–synthesis of current knowledge and a way forward. Glob Chang Biol. 2011;17(11):3392–404.
                              • 99. Ferreira V, Chauvet E. Future increase in temperature more than decrease in litter quality can affect microbial litter decomposition in streams. Oecologia. 2011;167(1):279–91. pmid:21461934
                              • 100. Suseela V, Conant RT, Wallenstein MD, Dukes JS. Effects of soil moisture on the temperature sensitivity of heterotrophic respiration vary seasonally in an old-field climate change experiment. Glob Chang Biol. 2012;18(1):336–48.
                              • 101. Ward SE, Orwin KH, Ostle NJ, Briones MJI, Thomson BC, Griffiths RI, et al. Vegetation exerts a greater control on litter decomposition than climate warming in peatlands. Ecology. 2015;96(1):113–23. pmid:26236896
                              • 102. Cheng L, Booker FL, Tu C, Burkey KO, Zhou L, Shew HD, et al. Arbuscular mycorrhizal fungi increase organic carbon decomposition under elevated CO2. Science. 2012;337(6098):1084–7. pmid:22936776
                              • 103. de Oliveira ED, Bramley H, Siddique KHM, Henty S, Berger J, Palta JA. Can elevated CO2 combined with high temperature ameliorate the effect of terminal drought in wheat? Funct Plant Biol. 2013;40(2):160–71.
                              • 104. Lindroth RL. Impacts of elevated atmospheric CO2 and O3 on forests: phytochemistry, trophic interactions, and ecosystem dynamics. J Chem Ecol. 2010;36(1):2–21. pmid:20054619
                              • 105. Rutledge S, Campbell DI, Baldocchi D, Schipper LA. Photodegradation leads to increased carbon dioxide losses from terrestrial organic matter. Glob Chang Biol. 2010;16(11):3065–74.
                              • 106. Varga B, Vida G, Varga-László E, Hoffmann B, Veisz O. Combined effect of drought stress and elevated atmospheric CO2 concentration on the yield parameters and water use properties of winter wheat (Triticum aestivum L.) genotypes. Journal of Agronomy and Crop Science. 2016.
                              • 107. Andresen LC, Michelsen A, Jonasson S, Schmidt IK, Mikkelsen TN, Ambus P, et al. Plant nutrient mobilization in temperate heathland responds to elevated CO2, temperature and drought. Plant Soil. 2010;328(1):381–96.
                              • 108. van Groenigen KJ, Qi X, Osenberg CW, Luo Y, Hungate BA. Faster decomposition under increased atmospheric CO2 limits soil carbon storage. Science. 2014;344(6183):508–9. pmid:24762538