Figure 1a: Litter inputs join SOM as light POM, which is largely composed of undecomposed litter fragments. As decomposition progresses, litter-derived SOM can more easily become incorporated into aggregates in heavy POM or microbial decomposition products and necromass can preferentially sorb to soil mineral surfaces as MAOM.1b: Roots and root-associated fungal symbionts can enhance both retention or loss of litter in light POM (top), heavy POM (middle), and MAOM (bottom) pools.
Living roots and their associated fungi alter SOM formation by sending C-rich exudates to the rhizosphere to enhance decomposition and acquire N (Bais et al., 2006; Grayston et al., 1997). However, a high degree of uncertainty remains in whether this increases or decreases soil C accumulation. In Figure 1b, we diagram potential hypotheses for how roots could alter litter loss from light POM and the accumulation of new heavy POM and MAOM through distinct mechanisms. First, root stimulation of microbial decomposition to mineralize soil N can increase the loss of unprotected light POM through the rhizosphere priming effect (Cheng et al., 2014). However, there is also evidence that roots and symbiotic fungi can outcompete saprotrophic microbes for resources like water and nutrients leading to the suppression of decomposition (Fernandez & Kennedy, 2016). Second, as litter inputs are transferred into more protected heavy POM, root ingrowth has the potential to both invade aggregates and increase the formation rate of new aggregates (Six et al., 2000). Finally, roots can enhance new MAOM formation by increasing the efficiency of microbial litter decomposition, resulting in greater microbial biomass production and the formation of microbial necromass (Liang et al., 2017). This necromass can associate with mineral surfaces and is the main precursor to MAOM in grassland ecosystems (Angst et al., 2021). However, roots may also deplete new, litter-derived MAOM as recent evidence suggests that roots can actively mine MAOM for nutrients (Jilling et al., 2021) and that root exudate compounds can displace MAOM from soil minerals (Keiluweit et al., 2015). As such, predicting whether roots will drive a net gain or loss of soil C is hindered by uncertainty in how roots impact SOM formation in these different pools.
The extent to which roots and mycorrhizal fungi facilitate SOM formation or loss in agricultural ecosystems may be modulated by fertilization. For example, some N-limited plants can dynamically shift C allocation belowground to root exudation and mycorrhizal symbionts to stimulate microbial decomposition in the rhizosphere and increase N acquisition (Brzostek et al., 2014; Kane et al., 2022). When N limitation is alleviated by fertilization, plants can also reduce belowground C allocation, suppressing SOM decomposition (Eastman et al., 2021; Frey et al., 2014). The degree to which fertilization alters SOM cycling also depends upon the activity of saprotrophic soil microbial decomposers. In contrast to plants, soil microbes are primarily understood to be energy, or carbon, limited (Soong et al., 2020). As such, organic fertilizer that contains C and N can prime microbial activity and decomposition relative to inorganic N fertilizer (Cui et al., 2022; Ndung’u et al., 2021). However, uncertainty remains in the extent to which the priming of microbial activity leads to net soil C losses by enhancing decomposition or net C gains by promoting the production of microbial necromass that can form MAOM. Collectively, the effect of fertilization on SOM formation depends upon the strength of plant-microbe interactions and the form of fertilizer applied, but the magnitude of this effect is uncertain.
Given the uncertainty above, our objectives were to: 1) determine how living roots and symbiotic fungi influence litter decomposition and SOM formation in distinct SOM pools and 2) assess how microbially-driven SOM formation is altered by fertilization. For the first objective, we assayed the net effect of the opposing hypotheses illustrated in Figure 1. For the second objective, we tested two hypotheses: (1) the effect of living roots on SOM formation would be strongest in unfertilized soil and (2) organic fertilizer would accelerate microbial decomposition and SOM cycling to a greater extent than inorganic fertilizer (SI Figure 1 ). To meet our objectives, we measured the effects of living roots and fungi on new SOM formation from isotopically enriched litter over one growing season. We incubated litter inputs in soil cores that were open to roots and fungal ingrowth (root), that excluded roots but were open to fungal ingrowth (fungal), or that excluded both roots and fungi (none) to quantify the effect of living roots and fungi on new SOM formation (SI Fig.2 ). We installed ingrowth cores in Miscanthus x giganteus (herein miscanthus) plots with different nutrient treatments to investigate the effect of soil N and C availability on how roots, mycorrhizal fungi, and saprotrophic microbes drive the transfer of litter C and N into light POM, heavy POM, and MAOM. We used the bioenergy feedstock crop miscanthus as a study system because it produces extensive root systems to overcome nutrient limitation (Dohleman & Long, 2009; Heaton et al., 2008) and because miscanthus agriculture typically increases SOM levels (Harris et al., 2015). Further, because bioenergy offers the potential to become a C neutral or C negative alternative to fossil fuels, it is particularly critical to investigate what drives SOM accumulation in these ecosystems (Hanssen et al., 2020).
We show that miscanthus roots increased litter decomposition but did not lead to a net C loss because roots enhanced the incorporation of litter C into heavy POM. Roots also selectively mobilized litter N from both POM pools. As such, roots can transfer C into a more persistent SOM pool while still enhancing N mining. These root effects did not depend on fertilization. However, organic fertilization enhanced microbial litter decomposition without increasing litter incorporation in MAOM.