Mycorrhizal roots slow the decay of belowground litters in a temperate hardwood forest.

There is increasing evidence that plant roots and mycorrhizal fungi, whether living or dead, play a central role in soil carbon (C) cycling. Root-mycorrhizal-microbial interactions can both suppress and enhance litter decay, with the net result dependent upon belowground nutrient acquisition strategies and soil nutrient availability. We measured the net effect of living roots and mycorrhizal fungi on the decay of dead roots and fungal hyphae in a hardwood forest dominated by either sugar maple (Acer saccharum) or white oak (Quercus alba) trees. Root and fungal litter were allowed to decompose within root-ingrowth bags and root-exclusion cores. In conjunction with root effects on decay, we assessed foraging responses and root induced changes in soil moisture, nitrogen (N) availability and enzyme activity. After 1 year, maple root production increased, and mycorrhizal fungal colonization decreased in the presence of decaying litter. In addition, we found that actively foraging roots suppressed the decay of root litter (- 14%) more than fungal litter (- 3%), and suppression of root decay was stronger for oak (- 20%) than maple roots (- 8%). Suppressive effects of oak roots on decay were greatest when roots also reduced soil N availability, which corresponded with reductions in hydrolytic enzyme activity and enhanced oxidative enzyme activities. These findings further our understanding of context-dependent drivers of root-mycorrhizal-microbial interactions and demonstrate that such interactions can play an underappreciated role in soil organic matter accumulation and turnover in temperate forests.

Redefining fine roots improves understanding of below-ground contributions to terrestrial biosphere processes.

Fine roots acquire essential soil resources and mediate biogeochemical cycling in terrestrial ecosystems. Estimates of carbon and nutrient allocation to build and maintain these structures remain uncertain because of the challenges of consistently measuring and interpreting fine-root systems. Traditionally, fine roots have been defined as all roots ≤ 2 mm in diameter, yet it is now recognized that this approach fails to capture the diversity of form and function observed among fine-root orders. Here, we demonstrate how order-based and functional classification frameworks improve our understanding of dynamic root processes in ecosystems dominated by perennial plants. In these frameworks, fine roots are either separated into individual root orders or functionally defined into a shorter-lived absorptive pool and a longer-lived transport fine-root pool. Using these frameworks, we estimate that fine-root production and turnover represent 22% of terrestrial net primary production globally - a c. 30% reduction from previous estimates assuming a single fine-root pool. Future work developing tools to rapidly differentiate functional fine-root classes, explicit incorporation of mycorrhizal fungi into fine-root studies, and wider adoption of a two-pool approach to model fine roots provide opportunities to better understand below-ground processes in the terrestrial biosphere.

The rhizosphere and hyphosphere differ in their impacts on carbon and nitrogen cycling in forests exposed to elevated CO2.

While multiple experiments have demonstrated that trees exposed to elevated CO2 can stimulate microbes to release nutrients from soil organic matter, the importance of root- versus mycorrhizal-induced changes in soil processes are presently unknown. We analyzed the contribution of roots and mycorrhizal activities to carbon (C) and nitrogen (N) turnover in a loblolly pine (Pinus taeda) forest exposed to elevated CO2 by measuring extracellular enzyme activities at soil microsites accessed via root windows. Specifically, we quantified enzyme activity from soil adjacent to root tips (rhizosphere), soil adjacent to hyphal tips (hyphosphere), and bulk soil. During the peak growing season, CO2 enrichment induced a greater increase of N-releasing enzymes in the rhizosphere (215% increase) than in the hyphosphere (36% increase), but a greater increase of recalcitrant C-degrading enzymes in the hyphosphere (118%) than in the rhizosphere (19%). Nitrogen fertilization influenced the magnitude of CO2 effects on enzyme activities in the rhizosphere only. At the ecosystem scale, the rhizosphere accounted for c. 50% and 40% of the total activity of N- and C-releasing enzymes, respectively. Collectively, our results suggest that root exudates may contribute more to accelerated N cycling under elevated CO2 at this site, while mycorrhizal fungi may contribute more to soil C degradation.

Changes in root architecture under elevated concentrations of CO2 and nitrogen reflect alternate soil exploration strategies.

Predicting the response of fine roots to increased atmospheric CO2 concentration has important implications for carbon (C) and nutrient cycling in forest ecosystems. Root architecture is known to play an important role in how trees acquire soil resources in changing environments. However, the effects of elevated CO2 on the fine-root architecture of trees remain unclear. We investigated the architectural response of fine roots exposed to 14 yr of CO2 enrichment and 6 yr of nitrogen (N) fertilization in a Pinus taeda (loblolly pine) forest. Root traits reflecting geometry, topology and uptake function were measured on intact fine-root branches removed from soil monoliths and the litter layer. CO2 enrichment resulted in the development of a fine-root pool that was less dichotomous and more exploratory under N-limited conditions. The per cent mycorrhizal colonization did not differ among treatments, suggesting that root growth and acclimation to elevated CO2 were quantitatively more important than increased mycorrhizal associations. Our findings emphasize the importance of architectural plasticity in response to environmental change and suggest that changes in root architecture may allow trees to effectively exploit larger volumes of soil, thereby pre-empting progressive nutrient limitations.

Root length, biomass, tissue chemistry and mycorrhizal colonization following 14 years of CO2 enrichment and 6 years of N fertilization in a warm temperate forest.

Root systems serve important roles in carbon (C) storage and resource acquisition required for the increased photosynthesis expected in CO2-enriched atmospheres. For these reasons, understanding the changes in size, distribution and tissue chemistry of roots is central to predicting the ability of forests to capture anthropogenic CO2. We sampled 8000 cm(3) soil monoliths in a pine forest exposed to 14 years of free-air-CO2-enrichment and 6 years of nitrogen (N) fertilization to determine changes in root length, biomass, tissue C : N and mycorrhizal colonization. CO2 fumigation led to greater root length (98%) in unfertilized plots, but root biomass increases under elevated CO2 were only found for roots <1 mm in diameter in unfertilized plots (59%). Neither fine root [C] nor [N] was significantly affected by increased CO2. There was significantly less root biomass in N-fertilized plots (19%), but fine root [N] and [C] both increased under N fertilization (29 and 2%, respectively). Mycorrhizal root tip biomass responded positively to CO2 fumigation in unfertilized plots, but was unaffected by CO2 under N fertilization. Changes in fine root [N] and [C] call for further study of the effects of N fertilization on fine root function. Here, we show that the stimulation of pine roots by elevated CO2 persisted after 14 years of fumigation, and that trees did not rely exclusively on increased mycorrhizal associations to acquire greater amounts of required N in CO2-enriched plots. Stimulation of root systems by CO2 enrichment was seen primarily for fine root length rather than biomass. This observation indicates that studies measuring only biomass might overlook shifts in root systems that better reflect treatment effects on the potential for soil resource uptake. These results suggest an increase in fine root exploration as a primary means for acquiring additional soil resources under elevated CO2.

Long-term dynamics of mycorrhizal root tips in a loblolly pine forest grown with free-air CO2 enrichment and soil N fertilization for 6 years.

Large-scale, long-term FACE (Free-Air CO2 enrichment) experiments indicate that increases in atmospheric CO2 concentrations will influence forest C cycling in unpredictable ways. It has been recently suggested that responses of mycorrhizal fungi could determine whether forest net primary productivity (NPP) is increased by elevated CO2 over long time periods and if forests soils will function as sources or sinks of C in the future. We studied the dynamic responses of ectomycorrhizae to N fertilization and atmospheric CO2 enrichment at the Duke FACE experiment using minirhizotrons over a 6 year period (2005-2010). Stimulation of mycorrhizal production by elevated CO2 was observed during only 1 (2007) of 6 years. This increased the standing crop of mycorrhizal tips during 2007 and 2008; during 2008, significantly higher mortality returned standing crop to ambient levels for the remainder of the experiment. It is therefore unlikely that increased production of mycorrhizal tips can explain the lack of progressive nitrogen limitations and associated increases in N uptake observed in CO2 -enriched plots at this site. Fertilization generally decreased tree reliance on mycorrhizae as tip production declined with the addition of nitrogen as has been shown in many other studies. Annual NPP of mycorrhizal tips was greatest during years with warm January temperatures and during years with cool spring temperatures. A 2 °C increase in average late spring temperatures (May and June) decreased annual production of mycorrhizal root tip length by 50%. This has important implications for ecosystem function in a warmer world in addition to potential for forest soils to sequester atmospheric C.

Sampling volume in root studies: the pitfalls of under-sampling exposed using accumulation curves.

Root systems are important for global models of below-ground carbon and nutrient cycling. Notoriously difficult sampling methods and the fractal distribution of root diameters in the soil make data being used in these models especially susceptible to error resulting from under-sampling. We applied the concept of species accumulation curves to root data to quantify the extent of under-sampling inherent to minirhizotron and soil coring sampling for both root uptake and carbon content studies. Based on differences in sample size alone, minirhizotron sampling missed approximately one third of the root diameters observed by soil core sampling. Sample volumes needed to encounter 90% of root diameters averaged 2481 cm(3) for uptake studies and 5878 cm(3) for root carbon content studies. These results show that small sample volumes encounter a non-representative sample of the overall root pool, and provide future guidelines for determining optimal sample volumes in root studies.

Increases in the flux of carbon belowground stimulate nitrogen uptake and sustain the long-term enhancement of forest productivity under elevated CO2.

The earth's future climate state is highly dependent upon changes in terrestrial C storage in response to rising concentrations of atmospheric CO2. Here we show that consistently enhanced rates of net primary production (NPP) are sustained by a C-cascade through the root-microbe-soil system; increases in the flux of C belowground under elevated CO2 stimulated microbial activity, accelerated the rate of soil organic matter decomposition and stimulated tree uptake of N bound to this SOM. This process set into motion a positive feedback maintaining greater C gain under elevated CO2 as a result of increases in canopy N content and higher photosynthetic N-use efficiency. The ecosystem-level consequence of the enhanced requirement for N and the exchange of plant C for N belowground is the dominance of C storage in tree biomass but the preclusion of a large C sink in the soil.

Soil incorporation of logging residue affects fine-root and mycorrhizal root-tip dynamics of young loblolly pine clones.

Loblolly pine (Pinus taeda L.) plantations cover a large geographic area of the southeastern USA and supply a large proportion of the nation's wood products. Research on management strategies designed to maximize wood production while also optimizing nutrient use efficiency and soil C sequestration is needed. We used minirhizotrons to quantify the effects of incorporating logging residues into soil on fine-root standing crop, production and mortality, and mycorrhizal root tips in young loblolly pine clones of contrasting ideotypes. Clone 93 is known to allocate more C to stem growth, while clone 32 allocates less C to stems and more to leaves. The relative allocation by these clones to support fine-root turnover is unknown. Clone 32 exhibited 37% more fine-root mortality than clone 93, which was mainly the result of a greater standing crop of fine roots. Fine-root standing crop in plots amended with logging residue was initially higher than control plots, but 2.5 years after planting, standing crop in control plots had exceeded that in mulched plots. Production of mycorrhizal root tips, on the other hand, was initially higher in control than mulched plots, but during the last 9 months of the study, mycorrhizal tip production was greater in mulched than control plots, especially for clone 93. As expected, turnover rate of fine roots was greater in surface soil (0-25 cm) compared with deeper (25-50 cm) soil and for small roots (< 0.4 mm diameter) compared with larger fine roots (0.4-2.0 mm diameter). Rates of fine-root turnover were similar in both clones. Organic matter additions reduced survivorship of individual roots and increased turnover rates of fine-root populations. Results indicate that management decisions should be tailored to fit the growth and allocation patterns of available clones.

Re-assessment of plant carbon dynamics at the Duke free-air CO(2) enrichment site: interactions of atmospheric [CO(2)] with nitrogen and water availability over stand development.

*The potential for elevated [CO(2)]-induced changes to plant carbon (C) storage, through modifications in plant production and allocation of C among plant pools, is an important source of uncertainty when predicting future forest function. Utilizing 10 yr of data from the Duke free-air CO(2) enrichment site, we evaluated the dynamics and distribution of plant C. *Discrepancy between heights measured for this study and previously calculated heights required revision of earlier allometrically based biomass determinations, resulting in higher (up to 50%) estimates of standing biomass and net primary productivity than previous assessments. *Generally, elevated [CO(2)] caused sustained increases in plant biomass production and in standing C, but did not affect the partitioning of C among plant biomass pools. Spatial variation in net primary productivity and its [CO(2)]-induced enhancement was controlled primarily by N availability, with the difference between precipitation and potential evapotranspiration explaining most interannual variability. Consequently, [CO(2)]-induced net primary productivity enhancement ranged from 22 to 30% in different plots and years. *Through quantifying the effects of nutrient and water availability on the forest productivity response to elevated [CO(2)], we show that net primary productivity enhancement by elevated [CO(2)] is not uniform, but rather highly dependent on the availability of other growth resources.

Irreconcilable differences: fine-root life spans and soil carbon persistence.

The residence time of fine-root carbon in soil is one of the least understood aspects of the global carbon cycle, and fine-root dynamics are one of the least understood aspects of plant function. Most recent studies of these belowground dynamics have used one of two methodological strategies. In one approach, based on analysis of carbon isotopes, the persistence of carbon is inferred; in the other, based on direct observations of roots with cameras, the longevity of individual roots is measured. We show that the contribution of fine roots to the global carbon cycle has been overstated because observations of root lifetimes systematically overestimate the turnover of fine-root biomass. On the other hand, isotopic techniques systematically underestimate the turnover of individual roots. These differences, by virtue of the separate processes or pools measured, are irreconcilable.

Increases in nitrogen uptake rather than nitrogen-use efficiency support higher rates of temperate forest productivity under elevated CO2.

Forest ecosystems are important sinks for rising concentrations of atmospheric CO(2). In previous research, we showed that net primary production (NPP) increased by 23 +/- 2% when four experimental forests were grown under atmospheric concentrations of CO(2) predicted for the latter half of this century. Because nitrogen (N) availability commonly limits forest productivity, some combination of increased N uptake from the soil and more efficient use of the N already assimilated by trees is necessary to sustain the high rates of forest NPP under free-air CO(2) enrichment (FACE). In this study, experimental evidence demonstrates that the uptake of N increased under elevated CO(2) at the Rhinelander, Duke, and Oak Ridge National Laboratory FACE sites, yet fertilization studies at the Duke and Oak Ridge National Laboratory FACE sites showed that tree growth and forest NPP were strongly limited by N availability. By contrast, nitrogen-use efficiency increased under elevated CO(2) at the POP-EUROFACE site, where fertilization studies showed that N was not limiting to tree growth. Some combination of increasing fine root production, increased rates of soil organic matter decomposition, and increased allocation of carbon (C) to mycorrhizal fungi is likely to account for greater N uptake under elevated CO(2). Regardless of the specific mechanism, this analysis shows that the larger quantities of C entering the below-ground system under elevated CO(2) result in greater N uptake, even in N-limited ecosystems. Biogeochemical models must be reformulated to allow C transfers below ground that result in additional N uptake under elevated CO(2).