Effects of fungicide treatments on mycorrhizal communities and carbon acquisition in the mixotrophic Pyrola japonica (Ericaceae).

Pyrola japonica, a member of the family Ericaceae, is a mixotroph that grows on forest floors and obtains carbon (C) from both its photosynthesis and its mycorrhizal fungi. Its mycorrhizal community is dominated by Russulaceae. However, the mechanism of its C acquisition and its flexibility are not well understood. Our aim was to assess the impact of disturbance of the mycorrhizal fungal communities on C acquisition by P. japonica. We repeatedly applied a fungicide (Benomyl) to soils around P. japonica plants in a broad-leaved forest of central Japan, in order to disturb fungal associates near roots. After fungicide treatment, P. japonica roots were collected and subjected to barcoding by next-generation sequencing, focusing on the ITS2 region. The rate of mycorrhizal formation and α-diversity did not significantly change upon fungicide treatments. Irrespective of the treatments, Russulaceae represented more than 80% of the taxa. Leaves and seeds of the plants were analysed for 13C stable isotope ratios that reflect fungal C gain. Leaf and seed δ13C values with the fungicide treatment were significantly lower than those with the other treatments. Thus the fungicide did not affect mycorrhizal communities in the roots, but disturbed mycorrhizal fungal pathways via extraradical hyphae, and resulted in a more photosynthetic behaviour of P. japonica for leaves and seeds.

Reconstruction of root systems in Cryptomeria japonica using root point coordinates and diameters.

MAIN CONCLUSION: We developed simple algorithms for reconstructing tree root system architecture using only the root point coordinate and diameter, which can be systematically obtained without digging up the root systems. Root system architecture (RSA) is strongly related to various root functions of the tree. The aim of this study was to develop a three-dimensional (3D) RSA model using systematically obtained information on root locations and root diameters at the locations. We excavated root systems of Cryptomeria japonica and systematically obtained XYZ coordinates and root diameters using a 10-cm grid. We clarified the patterns of the root point connections and developed a reconstructed root system model. We found that the root diameters farther from the stump centre are smaller. Additionally, we found that the root lengths of the segments running between the base and the connected root point were smaller than those of other root segments, and the inner angle between the base and the stump and between the base and the connected root point was narrower than for the other pairs. The new RSA model developed according to these results had average accuracies of 0.64 and 0.80 for estimates of total volume and length, respectively. The developed model can estimate 3D RSA using only root point data, which can be obtained without digging up root systems. This suggests a wide applicability of this model in root function evaluation.

Intraspecific variation in fine root respiration and morphology in response to in situ soil nitrogen fertility in a 100-year-old Chamaecyparis obtusa forest.

Soil N fertility has an effect on belowground C allocation, but the physiological and morphological responses of individual fine root segments to variations in N availability under field conditions are still unclear. In this study, the direction and magnitude of the physiological and morphological function of fine roots in response to variable in situ soil N fertility in a forest site were determined. We measured the specific root respiration (Rr) rate, N concentration and morphology of fine root segments with 1-3 branching orders in a 100-year-old coniferous forest of Chamaecyparis obtusa. Higher soil N fertility induced higher Rr rates, root N concentration, and specific root length (SRL), and lower root tissue density (RTD). In all fertility levels, the Rr rates were significantly correlated positively with root N and SRL and negatively with RTD. The regression slopes of respiration with root N and RTD were significantly higher along the soil N fertility gradient. Although no differences in the slopes of Rr and SRL relationship were found across the levels, there were significant shifts in the intercept along the common slope. These results suggest that a contrasting pattern in intraspecific relationships between specific Rr and N, RTD, and SRL exists among soils with different N fertility. Consequently, substantial increases in soil N fertility would exert positive effects on organ-scale root performance by covarying the Rr, root N, and morphology for their potential nutrient and water uptake.

Contrasting patterns of nitrogen release from fine roots and leaves driven by microbial communities during decomposition.

Leachate from decaying root and leaf litter plays crucial roles in soil biogeochemical processes in forest ecosystems. Unlike for leaf litter, however, the chemical composition and microbial community of root litter leachate are poorly understood. We hypothesized that both leachate nitrogen (N) composition and microbial communities differ between plant organs and decomposition stages and that leachate composition affects microbial community composition. We conducted a 2.5-year laboratory incubation using root and leaf substrate from Cryptomeria japonica and Chamaecyparis obtusa. We monitored the N forms released and used metabarcoding to characterize the microbial communities. Leachate N accounted for 40 % and 30 % of net N losses from C. japonica and C. obtusa roots, respectively; the remainder was probably lost in gaseous forms. In contrast, leaves absorbed N during the incubation regardless of tree species. The predominant N form in root leachate was nitrate (NO3-); cumulative NO3- quantity was 22.6 and 25.5 times greater in root than in leaf leachate for C. japonica and C. obtusa, respectively. A nitrifying bacterium was selected as the indicator taxon in root substrates, whereas many families of N-fixing bacteria were selected in leaf substrates. At the end of the incubation period, bacterial taxonomic diversity was high in both organs from both tree species, ranging from 177 to 339 taxa and increasing with time. However, fungal diversity was low for both organs (72 to 155 taxa). Shifts in bacterial community structure were related to NO3- concentration and leachate pH, whereas shifts in fungal community structure were related to leachate pH. These results suggest that the contrasting N dynamics of root and leaf substrates are strongly affected by the characteristics of and the microbes recruited by their leachates. Understanding organ-specific litter N dynamics is indispensable for predicting N cycling for optimal management of forest ecosystems in a changing world.

Leachate from fine root litter is more acidic than leaf litter leachate: A 2.5-year laboratory incubation.

Some tree species increase fine root production under soil acidification, thus changing the balance of litter input from leaves and roots. Litter leaches a significant amount of acidic materials during its decomposition, which might facilitate soil acidification. In this context, we focused on dissolved organic matter (DOM) as the major component of acidic materials. We hypothesized that both the quality and quantity of DOM, which control its function (i.e., proton supply), differ between leaf and root litter. To test this hypothesis, we conducted a 2.5-year laboratory incubation experiment using fresh fine roots and fresh green leaves as litter of two coniferous species (Cryptomeria japonica and Chamaecyparis obtusa) and investigated the leachate pH and DOM composition based on the optical properties. After the early stage of decomposition when flash leaching of DOM converged, the amount of dissolved organic carbon (DOC) leached from roots increased again and leachate pH declined. In contrast, DOC concentrations continued to decrease in leaf leachates during the incubation period, and the pH decrease was not as striking as that of root leachates. Optical properties (ultraviolet visible absorption and fluorescence) of DOM revealed that humic-like substances in DOM played a central role in the acidic pH of root leachates. The total amount of protons released from roots of C. japonica and C. obtusa is about 13 and 18 times higher, respectively, than that from leaves. These results imply that the increase of fine root biomass may induce a positive plant-soil feedback in acidic soils, affecting soil biogeochemical functions of terrestrial ecosystems.

Calculation procedures to estimate fine root production rates in forests using two-dimensional fine root data obtained by the net sheet method.

Several recent studies have used the net sheet method to estimate fine root production rates in forest ecosystems, wherein net sheets are inserted into the soil and fine roots growing through them are observed. Although this method has advantages in terms of its easy handling and low cost, there are uncertainties in the estimates per unit soil volume or unit stand area, because the net sheet is a two-dimensional material. Therefore, this study aimed to establish calculation procedures for estimating fine root production rates from two-dimensional fine root data on net sheets. This study was conducted in a hinoki cypress (Chamaecyparis obtusa (Sieb. & Zucc.) Endl.) stand in western Japan. We estimated fine root production rates in length and volume from the number (RN) and cross-sectional area (RCSA) densities, respectively, for fine roots crossing the net sheets, which were then converted to dry mass values. For these calculations, we used empirical regression equations or theoretical equations between the RN or RCSA densities on the vertical walls of soil pits and fine root densities in length or volume, respectively, in the soil, wherein the theoretical equations assumed random orientation of the growing fine roots. The estimates of mean fine root (diameter <1 mm) production rates were ∼80-100 g m-2 year-1 using the empirically obtained regression equations, whereas those from the theoretical equations were ∼40-50 g m-2 year-1. The difference in the estimates was attributed to larger slope values of the empirical regression equations than those of the theoretical equations, suggesting that fine root orientation was not random in our study site. In light of these results, we concluded that fine root production rates were successfully estimated from two-dimensional fine root data on the net sheets using these calculation procedures, with the empirical regression equations reflecting fine root orientation in the study site.