Increased belowground tree carbon allocation in a mature mixed forest in a dry versus a wet year.

Tree species differ in their carbon (C) allocation strategies during environmental change. Disentangling species-specific strategies and contribution to the C balance of mixed forests requires observations at the individual tree level. We measured a complete set of C pools and fluxes at the tree level in five tree species, conifers and broadleaves, co-existing in a mature evergreen mixed Mediterranean forest. Our study period included a drought year followed by an above-average wet year, offering an opportunity to test the effect of water availability on tree C allocation. We found that in comparison to the wet year, C uptake was lower in the dry year, C use was the same, and allocation to belowground sinks was higher. Among the five major C sinks, respiration was the largest (ca. 60%), while root exudation (ca. 10%) and reproduction (ca. 2%) were those that increased the most in the dry year. Most trees relied on stored starch for maintaining a stable soluble sugars balance, but no significant differences were detected in aboveground storage between dry and wet years. The detailed tree-level analysis of nonstructural carbohydrates and δ13 C dynamics suggest interspecific differences in C allocation among fluxes and tissues, specifically in response to the varying water availability. Overall, our findings shed light on mixed forest physiological responses to drought, an increasing phenomenon under the ongoing climate change.

Low light-regulated intramolecular disulfide fine-tunes the role of PTOX in Arabidopsis.

Disulfide-based regulation links the activity of numerous chloroplast proteins with photosynthesis-derived redox signals. The plastid terminal oxidase (PTOX) is a thylakoid-bound plastoquinol oxidase that has been implicated in multiple roles in the light and in the dark, which could require different levels of PTOX activity. Here we show that Arabidopsis PTOX contains a conserved C-terminus domain (CTD) with cysteines that evolved progressively following the colonization of the land by plants. Furthermore, the CTD contains a regulatory disulfide that is in the oxidized state in the dark and is rapidly reduced, within 5 min, in low light intensity (1-5 µE m-2 sec-1 ). The reduced PTOX form in the light was reoxidized within 15 min after transition to the dark. Mutation of the cysteines in the CTD prevented the formation of the oxidized form. This resulted in higher levels of reduced plastoquinone when measured at transition to the onset of low light. This is consistent with the reduced state of PTOX exhibiting diminished PTOX oxidase activity under conditions of limiting PQH2 substrate. Our findings suggest that AtPTOX-CTD evolved to provide light-dependent regulation of PTOX activity for the adaptation of plants to terrestrial conditions.

Asymmetric belowground carbon transfer in a diverse tree community.

Mycorrhizal fungi can colonize multiple trees of a single or multiple taxa, facilitating bidirectional exchange of carbon between trees. Mycorrhiza-induced carbon transfer was shown in the forest, but it is unknown whether carbon is shared symmetrically among tree species, and if not, which tree species are better donors and which are better recipients. Here, we test this question by investigating carbon transfer dynamics among five Mediterranean tree species in a microcosm system, including both ectomycorrhizal (EM) and arbuscular (AM) plants. Trees were planted together in "community boxes" using natural soil from a mixed forest plot that serves as a habitat for all five tree species and their native mycorrhizal fungi. In each box, only the trees of a single species were pulse-labelled with 13 CO2 . We found that carbon transfer was asymmetric, with oak being a better donor, and pistacia and cypress better recipients. Shared mycorrhizal species may have facilitated carbon transfer, but their diversity did not affect the amount, nor timing, of the transfer. Overall, our findings in a microcosm system expose rich, but hidden, belowground interactions in a diverse population of trees and mycorrhizal fungi. The asymmetric carbon exchange among cohabiting tree species could potentially contribute to forest resilience in an uncertain future.

Share the wealth: Trees with greater ectomycorrhizal species overlap share more carbon.

The mutualistic symbiosis between forest trees and ectomycorrhizal fungi (EMF) is among the most ubiquitous and successful interactions in terrestrial ecosystems. Specific species of EMF are known to colonize specific tree species, benefitting from their carbon source, and in turn, improving their access to soil water and nutrients. EMF also form extensive mycelial networks that can link multiple root-tips of different trees. Yet the number of tree species connected by such mycelial networks, and the traffic of material across them, are just now under study. Recently we reported substantial belowground carbon transfer between Picea, Pinus, Larix and Fagus trees in a mature forest. Here, we analyze the EMF community of these same individual trees and identify the most likely taxa responsible for the observed carbon transfer. Among the nearly 1,200 EMF root-tips examined, 50%-70% belong to operational taxonomic units (OTUs) that were associated with three or four tree host species, and 90% of all OTUs were associated with at least two tree species. Sporocarp 13 C signals indicated that carbon originating from labelled Picea trees was transferred among trees through EMF networks. Interestingly, phylogenetically more closely related tree species exhibited more similar EMF communities and exchanged more carbon. Our results show that belowground carbon transfer is well orchestrated by the evolution of EMFs and tree symbiosis.

ACHT4-driven oxidation of APS1 attenuates starch synthesis under low light intensity in Arabidopsis plants.

The regulatory mechanisms that use signals of low levels of reactive oxygen species (ROS) could be obscured by ROS produced under stress and thus are better investigated under homeostatic conditions. Previous studies showed that the chloroplastic atypical thioredoxin ACHT1 is oxidized by 2-Cys peroxiredoxin (2-Cys Prx) in Arabidopsis plants illuminated with growth light and in turn transmits a disulfide-based signal via yet unknown target proteins in a feedback regulation of photosynthesis. Here, we studied the role of a second chloroplastic paralog, ACHT4, in plants subjected to low light conditions. Likewise, ACHT4 reacted in planta with 2-Cys Prx, indicating that it is oxidized by a similar disulfide exchange reaction. ACHT4 further reacted uniquely with the small subunit (APS1) of ADP-glucose pyrophosphorylase (AGPase), the first committed enzyme of the starch synthesis pathway, suggesting that it transfers the disulfides it receives from 2-Cys Prx to APS1 and turns off AGPase. In accordance, ACHT4 participated in an oxidative signal that quenched AGPase activity during the diurnal transition from day to night, and also in an attenuating oxidative signal of AGPase in a dynamic response to small fluctuations in light intensity during the day. Increasing the level of expressed ACHT4 or of ACHT4ΔC, a C terminus-deleted form that does not react with APS1, correspondingly decreased or increased the level of reduced APS1 and decreased or increased transitory starch content. These findings imply that oxidative control mechanisms act in concert with reductive signals to fine tune starch synthesis during daily homeostatic conditions.

Inter-generational consistency of the ectomycorrhizal fungal community in a mixed pine-cedar post-fire stand.

The mutualistic interaction between trees and ectomycorrhizal fungi (EMF) can have a major effect on forest dynamics and specifically on seedling establishment. Here, we compared the EMF community composition associated with the roots of young saplings and mature trees of two co-habiting Pinaceae: Pinus halepensis and Cedrus deodara growing together in a post-fire forest plot, using fungal ITS metabarcoding. We found that the differences in the EMF community between the two sapling groups were mostly attributed to changes in the relative abundance of specific fungal species with little species turnover. Specifically, Tomentella showed high abundance on pine roots while Tuber, Russula and Sebacina were more common on the roots of cedars. The physical proximity to a specific host species was correlated with the EMF community composition of young saplings. Specifically, regardless of the sapling's own identity, the roots of saplings growing next to mature cedars had higher abundance of Tuber species while Tomentella coerulea, Russula densifolia and Tuber nitidum dominated saplings next to mature pines. Cedar saplings' shoot structure was correlated with a specific EMF species. Overall, these results suggest that when germinating next to mature trees, the EMF community of saplings could be determined by extrinsic factors such as the small-scale distribution of mature trees in the forest.

Belowground carbon transfer across mycorrhizal networks among trees: Facts, not fantasy.

The mycorrhizal symbiosis between fungi and plants is among the oldest, ubiquitous and most important interactions in terrestrial life on Earth. Carbon (C) transfer across a common mycorrhizal network (CMN) was demonstrated over half a century ago in the lab ( Reid & Woods, 1969), and later in the field ( Simard et al., 1997a). Recent years have seen ample progress in this research direction, including evidence for ecological significance of carbon transfer ( Klein et al., 2016). Furthermore, specific cases where the architecture of mycorrhizal networks have been mapped ( Beiler et al., 2015) and CMN-C transfer from mature trees to seedlings has been demonstrated ( Orrego, 2018) have suggested that trees in forests are more connected than once thought ( Simard, 2021). In a recent Perspective, Karst et al. (2023) offered a valuable critical review warning of over-interpretation and positive citation bias in CMN research. It concluded that while there is evidence for C movement among plants, the importance of CMNs remains unclear, as noted by others too ( Henriksson et al., 2023). Here we argue that while some of these claims are justified, factual evidence about belowground C transfer across CMNs is solid and accumulating.

Enhanced root exudation of mature broadleaf and conifer trees in a Mediterranean forest during the dry season.

Root exudates are part of the rhizodeposition process, which is the major source of soil organic carbon (C) released by plant roots. This flux of C is believed to have profound effects on C and nutrient cycling in ecosystems. The quantity of root exudates depends on the plant species, the period throughout the year, and external biotic and abiotic factors. Since root exudates of mature trees are difficult to collect in field conditions, very little is known about their flux, especially in water-limited ecosystems, such as the seasonally hot and dry Mediterranean maquis. Here, we collected exudates from DNA-identified roots in the forest from the gymnosperm Cupressus sempervirens L. and the evergreen angiosperm Pistacia lentiscus L. by 48-h incubations on a monthly temporal resolution throughout the year. We examined relationships of the root exudate C flux to abiotic parameters of the soil (water content, water potential, temperature) and atmosphere (vapor pressure deficit, temperature). We also studied relationships to C fluxes through the leaves as indicators of tree C balance. Root exudation rates varied significantly along the year, increasing from 6 µg C cm -2 root day-1 in both species in the wet season to 4- and 11-fold rates in Pistacia and Cupressus, respectively, in the dry season. A stepwise linear mixed-effects model showed that the three soil parameters were the most influential on exudation rates. Among biotic factors, there was a significant negative correlation of exudation rate with leaf assimilation in Cupressus and a significant negative correlation with leaf respiration in Pistacia. Our observation of enhanced exudation flux during the dry season indicates that exudation dynamics in the field are less sensitive to the low tree C availability in the dry season. The two key Mediterranean forest species seem to respond to seasonal changes in the rhizosphere such as drying and warming, and therefore invest C in the rhizosphere under seasonal drought.

Starch biosynthesis by AGPase, but not starch degradation by BAM1/3 and SEX1, is rate-limiting for CO2 -regulated stomatal movements under short-day conditions.

Starch in guard cells functions in osmoregulation during stomatal movements. Starch metabolism is controlled by the circadian clock. We investigated the role of starch metabolism in stomatal responses to CO2 under different photoperiodic conditions. Guard cell starch levels correlate with low/high [CO2 ] exposure. Starch biosynthesis-deficient AGPase (ADG1) mutants but, unexpectedly, not the starch degradation-deficient BAM1, BAM3, and SEX1 mutants alone, are rate-limiting for stomatal conductance responses to [CO2 ]-shifts. Interestingly, AGPase is rate-limiting solely under short- but not long-day conditions. These findings suggest a model of enhanced AGPase activity in guard cells under short days such that starch biosynthesis becomes rate-limiting for CO2 -induced stomatal closing.