Group VII Ethylene Response Factors (ERF-VII) are plant-specific transcription factors (TFs) known for their role in the activation of hypoxia-responsive genes under low oxygen stress but also in plant endogenous hypoxic niches. However, their function in the microaerophilic nitrogen-fixing nodules of legumes has not yet been investigated. We investigated regulation and the function of the two Medicago truncatula ERF-VII TFs ( MtERF74 and MtERF75) in roots and nodules, MtERF74 and MtERF75 in response to hypoxia stress and during the nodulation process using an RNA interference strategy and targeted proteolysis of MtERF75. Knockdown of MtERF74 and MtERF75 partially blocked the induction of hypoxia-responsive genes in roots exposed to hypoxia stress. In addition, a significant reduction in nodulation capacity and nitrogen fixation activity was observed in mature nodules of double knockdown transgenic roots. Overall, the results indicate that MtERF74 and MtERF75 are involved in the induction of MtNR1 and Pgb1.1 expression for efficient Phytogb-NO respiration in the nodule.

Plant roots interact with rhizosphere microbes to accelerate soil organic matter (SOM) mineralization and promote nutrient acquisition. Root-mediated changes in SOM turnover largely depend on root-C input and soil nutrient availability. Hence, interspecific competition and nutrient uptake dynamics over plant development stages as well as spatiotemporal variability in C input may modify SOM turnover. To investigate the effect of intraspecific competition on SOM mineralization at three growth stages (heading, flowering and ripening), we grew maize (C4 plant) under three planting densities on a C3 soil. 13C-natural abundance and 15N-pool dilution were applied in situ to determine C- and N-mineralization rates. Soil C- and N-mineralization rates were tightly coupled and peaked at maize flowering. However, the C-to-N-mineralization ratio increased with N, indicating that microbes mineralize N-rich components to mine SOM for N. Furthermore, intraspecific competition did not affect root biomass; instead, plants shaped root morphology towards higher specific root length as an efficient strategy competing for nutrient. Hence, root morphologic traits rather than root biomass per se were positively related to C- and N-mineralization. Overall, plant competition for nutrients controlled the intensity and mechanisms of soil C- and N-turnover by the adaptation of root traits and nutrient depletion.

Little is known about the sources and age of C respired from tree roots. Previous research in tree stems has identified two functional pools of non-structural carbohydrates (NSC): an ‘active’ pool supplied directly from canopy photo-assimilates that supports metabolism and a ‘stored’ pool used when fresh C supplies are limited. We compared the C isotope composition of water soluble NSC and respired CO2 for aspen roots (Populus tremula hybrids) that were cut off fresh C supply via stem-girdling and prolonged incubation of excised roots. We used bomb radiocarbon to estimate the time elapsed since C fixation for respired CO2, water-soluble C, and structural α-cellulose. While freshly excised roots respired CO2 with mean age <1 yr, within a week the age increased to 1.6-2.9 yr. Freshly excised roots from trees girdled ~3 months previously had similar respiration rates and NSC stocks as un-girdled trees, but respired older C (~1.2 yr). We estimate the NSC in girdled roots must be replaced 5-7 times by reserves remobilized from root-external sources. Using a mixing model and observed correlations between Δ14C of water-soluble C and α-cellulose, we estimate ~30% of C is ‘active’ (~5 mg C g-1).

Triose-phosphate utilization (TPU) limits the maximum rate at which plants can photosynthesize. However, TPU is almost never found to be limiting photosynthesis under ambient conditions for plants. This, along with previous results showing adaptability of TPU at low temperature, suggest that TPU capacity is regulated to be just above the photosynthetic rate achievable under the prevailing conditions. A set of experiments were performed to study the adaptability of TPU capacity when plants are acclimated to elevated CO 2 concentrations. Plants held at 1500 ppm CO 2 were initially TPU limited. After 30 hours they no longer exhibited TPU limitations but they did not elevate their TPU capacity. Instead, the maximum rates of carboxylation and electron transport declined. A timecourse of regulatory responses was established. A step increase of CO 2 first caused PSI to be oxidized but after 40 s both PSI and PSII had excess electrons as a result of acceptor-side limitations. Electron flow to PSI slowed and the proton motive force increased. Eventually, non-photochemical quenching reduced electron flow sufficiently to balance the TPU limitation. Over several minutes rubisco deactivated contributing to regulation of metabolism to overcome the TPU limitation.

Salt stress is a major limiting factor that severely affects the survival and growth of crops. It is important to understand the salt tolerance ability of Brassica napus and explore the underlying related genetic resources. We used a high-throughput phenotyping platform to quantify 2,111 image-based traits (i-traits) of a natural population under 3 different salt stress conditions and an intervarietal substitution line (ISL) population under 9 different stress conditions to monitor and evaluate the salt stress tolerance of B. napus over time. We finally identified 928 high-quality i-traits associated with the salt stress tolerance of B. napus. Moreover, we mapped the salt stress-related loci in the natural population via a genome-wide association study (GWAS) and performed a linkage analysis associated with the ISL population, respectively. The results revealed 234 candidate genes associated with salt stress response, and two novel candidate genes, BnCKX5 and BnERF3, were experimentally verified to regulate the salt stress tolerance of B. napus. This study demonstrates the feasibility of using high-throughput phenotyping-based QTL mapping to accurately and comprehensively quantify i-traits associated with B. napus. The mapped loci could be used for genomics-assisted breeding to genetically improve the salt stress tolerance of B. napus.

The linkage of stomatal behavior with photosynthetic processes is critical to understanding water and carbon cycles under global change. The slope ( g1) of stomatal conductance ( gs) versus CO 2 assimilation ( Anet) serves as a proxy of the marginal water cost of carbon acquisition and the trade-off between carbon gain and water loss. Here we use g1 to assess species differences in the response of stomatal behavior to experimental climate change manipulations, asking whether generalizable patterns exist across species and climate contexts. A total of 17,727 Anet- gs measurements made in a long-term open-air experiment under ambient and +3.3°C warming, and ambient and ~40% summer rainfall reduction provided > 2,700 estimates of g1 across 21 boreal and temperate tree species. All species became more conservative in their water use (lower g1) in warming and/or reduced rainfall treatments because of lower soil moisture. In contrast to these phenotypic responses, species from warmer and drier habitats tended to have slightly higher g1 and to be the least sensitive to the decrease in soil water. Overall, both warming and rainfall reduction consistently made stomatal behavior more conservative in terms of water loss per unit carbon gain across 21 species and a decade of experimental observation.

Long non-coding RNAs (lncRNAs) have been considered to be important regulators of gene expression in a range of biological processes in plants. A large number of lncRNAs have been identified in plants. However, most of their biological functions still remain to be determined. Here, we identified total 3 004 lncRNAs in cassava under normal or cold-treated conditions from Iso-seq data. We further characterized a lincRNA, CRIR1, as a novel positive regulator of the plant response to cold stress. CRIR1 can be significantly induced by cold treatment. Overexpression of CRIR1 in cassava enhanced the cold tolerance of transgenic plants. Transcriptome analysis demonstrated that CRIR1 regulates a range of cold stress-related genes in a CBF-independent pathway. We further found that CRIR1 RNA can interact with MeCSP5, a homolog of the cold shock protein that acts as RNA chaperones, indicating that CRIR1 may recruit MeCSP5 to improve the translation efficiency of mRNA. In summary, our study greatly extends the repertoire of lncRNAs in plants as well as its responding to cold stress. Moreover, it reveals a sophisticated mechanism by which CRIR1 regulates plant cold stress response by modulating the expression of stress-responsive genes and increasing the translational yield.

The regulation of protein synthesis plays an important role in growth and development in all organisms. Upstream open reading frames (uORFs) are commonly found in eukaryotic mRNA transcripts and typically attenuate the translation of associated downstream main ORFs (mORFs). Conserved peptide uORFs (CPuORFs) are a rare subset of uORFs, some of which have been shown to conditionally regulate translation by ribosome stalling. Here we identify three Arabidopsis CPuORFs of ancient origin that regulate translation of any downstream ORF, in response to agriculturally significant environmental signals: heat stress and water limitation. We provide evidence that different sequence classes of CPuORF stall ribosomes during different phases of translation and show that plant CPuORFs act as environmental sensors that can be utilised as inducible regulators of translation with broad application.

Water inside plants forms a continuous chain from water in soils to the water evaporating from leaf surfaces. Failures in this chain result in reduced transpiration and photosynthesis and these failures are caused by soil drying and/or cavitation-induced xylem embolism. Xylem embolism and plant hydraulic failure share a number of analogies to “catastrophe theory” in dynamical systems. These catastrophes are often represented in the physiological and ecological literature as tipping points or alternative stable states when control variables exogenous (e.g. soil water potential) or endogenous (e.g. leaf water potential) to the plant are allowed to slowly vary. Here, plant hydraulics viewed from the perspective of catastrophes at multiple spatial scales is considered with attention to bubble expansion (i.e. cavitation), organ-scale vulnerability to embolism, and whole-plant biomass as a proxy for transpiration and hydraulic function. The hydraulic safety-efficiency tradeoff, hydraulic segmentation and maximum plant transpiration are examined using this framework. Underlying mechanisms for hydraulic failure at very fine scales such as pit membranes, intermediate scales such as xylem network properties and at larger scales such as soil-tree hydraulic pathways are discussed. Lacunarity areas in plant hydraulics are also flagged where progress is urgently needed.

To understand the growth response to drought, we performed a proteomics study in the leaf growth zone of maize (Zea mays L.) seedlings and functionally characterized the role of starch biosynthesis in the regulation of growth, photosynthesis and antioxidant capacity, using the shrunken2 mutant (sh2), defective in ADP-glucose pyrophosphorylase. Drought induced differential expression of 284 proteins overrepresented for photosynthesis, amino acids, sugar and starch metabolism, and redox-regulation. Changes in protein levels correlated with enzyme activities (increased ATP synthase, cysteine synthase, starch synthase, RuBisCo, peroxiredoxin, glutaredoxin, thioredoxin and decreased triosephosphate isomerase, ferredoxin, cellulose synthase activities, respectively) and metabolite concentrations (increased ATP, cysteine, glycine, serine, starch, proline and decreased cellulose levels). The sh2 mutant had a reduced ability to increase starch levels under drought conditions, causing soluble sugar starvation at the end of the night and impaired leaf growth. Increased RuBisCo activity and pigment concentrations observed in WT in response to drought were lacking in the mutant, which suffered more oxidative damage and recovered more slowly after re-watering. These results demonstrate that starch biosynthesis plays a crucial role in maintaining leaf growth under drought stress and facilitates enhanced carbon acquisition upon recovery.

N-terminal cysteine oxidases (NCOs) use molecular oxygen to oxidize the amino-terminal cysteine of specific proteins, thereby initiating the proteolytic N-degron pathway. To expand the characterization of the plant family of NCOs (PCOs), we performed a phylogenetic analysis across different taxa in terms of sequence similarity and transcriptional regulation. Based on this survey, we propose a distinction of PCOs into two main groups. A-type PCOs are conserved across all plant species and are generally unaffected at the mRNA level by oxygen availability. Instead, B-type PCOs differentiated in spermatophytes to acquire transcriptional regulation in response to hypoxia. The inactivation of two A-type PCOs in Arabidopsis thaliana, PCO4 and PCO5, is sufficient to activate the anaerobic response in young seedlings, whereas the additional removal of B-type PCOs leads to a stronger induction of anaerobic genes and impairs plant growth and development. Our results show that both PCO types are required to regulate the anaerobic response in angiosperms. Therefore, while it is possible to distinguish two clades within the PCO family, we conclude that they all contribute to restrain the anaerobic transcriptional program in normoxic conditions and together generate a molecular switch to toggle the hypoxic response.

Several species of soil free-living saprotrophs can sometimes establish biotrophic symbiosis with plants, but the basic biology of this association remains largely unknown. Here, we investigate the symbiotic interaction between a common soil saprotroph, Clitopilus hobsonii (Agaricomycetes), and the American sweetgum (Liquidambar styraciflua). Notably, the colonized root cortical cells contain numerous microsclerotia-like structures. Fungal colonization led to increased plant growth and facilitated potassium uptake, particularly under potassium limitation (0.05 mM K+). The expression of plant genes related to potassium uptake is not altered during symbiosis, whereas the transcripts of three fungal genes encoding ACU, HAK, and SKC involved in K+ nutrition is found in colonized roots. We confirmed the K+ influx activities by expressing the ChACU and ChSKC genes into a yeast K+-uptake-defective mutant. Upregulation of the ChACU under 0.05 mM K+ and no K+ conditions was demonstrated in planta and in vitro compared to normal condition (5 mM K+). In addition, colonized plants displayed a larger accumulation of soluble sugars under 0.05 mM K+. The present study highlights that potassium limitation promotes this novel tree-fungus symbiosis mainly through a reciprocal transfer of additional carbon and potassium to both partners, and the role of dual soil saprotroph/symbiotroph in tree nutrition.

Human activity and natural processes have led to widespread dissemination of metals and metalloids, many of which are toxic and have a negative impact on agronomic production. Roots, as the first point of contact, are essential in endowing plants with tolerance to excess metal(loid) in the soil. The most important root responses include: adaptation of transport processes that affect uptake, efflux and long distance transport of metal(loid)s; metal(loid) detoxification within root cells via conjugation to thiol rich compounds and subsequent sequestration in the vacuole; plasticity in root architecture; the presence of bacteria and fungi in the rhizosphere that impact on metal(loid) bioavailability; the role of root exudates. In this review we will provide details on these processes and assess their relevance for the detoxification of arsenic, cadmium, mercury and zinc. Furthermore, we will assess if any of these methodologies has been tested in field conditions and whether they are effective in terms of improving crop metal(loid) tolerance.

Carbonate-rich soils limit plant performance and crop production. Previously, local adaptation to carbonated soils was detected in wild Arabidopsis thaliana accessions, allowing the selection of two demes with contrasting phenotypes: A1 (carbonate tolerant, c+) and T6 (carbonate sensitive, c-). Here, A1 (c+) and T6 (c-) seedlings were grown hydroponically under control (pH 5.9) and bicarbonate conditions (10 mM NaHCO 3, pH 8.3) to obtain ionomic profiles and conduct transcriptomic analysis. In parallel, A1 (c+) and T6 (c-) parental lines and their progeny were cultivated on carbonated soil to evaluate fitness and segregation patterns. To understand the genetic architecture beyond the contrasted phenotypes a bulk segregant analysis sequencing (BSA-Seq) was performed. Transcriptomics revealed 208 root and 2503 leaf differentially expressed genes (DEGs) in A1 (c+) vs T6 (c-) comparison under bicarbonate stress, mainly involved in iron, nitrogen and carbon metabolism, hormones, and glycosylates biosynthesis. Based on A1 (c+) and T6 (c-) genome contrasts and BSA-Seq analysis, 69 genes were associated with carbonate tolerance. Comparative analysis of genomics and transcriptomics discovered a final set of 18 genes involved in bicarbonate stress responses that may have relevant roles in soil carbonate tolerance.

Translocation of metabolites between different plant species provides important hints in understanding the fate of bioactive root exudates. In the present study, targeted and untargeted mass spectrometry-based metabolomics was applied to elucidate the transfer of bioactive compounds between rye and several crops and weed species. Our results demonstrated that benzoxinoids (BXs) synthesized by rye were taken up by roots of neighboring plant species and translocated into their shoots. Furthermore, we showed roots of the rye plant took up compounds originating from neighboring plants. Among the compounds taken up by rye roots, wogonin was detected in the rye shoot, which indicates the root-to-shoot translocation of this compound. Elucidating the transfer of bioactive compounds between plants is essential for understanding plant-plant interactions, developing natural pesticides and understanding their modes of action.

Capparis odoratissima is a tree species native to semi-arid environments of the northern coast of South America where low soil water availability coexists with frequent nighttime fog. A previous study showed that water applied to leaf surfaces enhanced leaf hydration, photosynthesis, and growth, but the mechanisms of foliar water uptake are unknown. Here we combine detailed anatomical evaluations with water and dye uptake experiments in the laboratory, and use immunolocalization of pectin and arabinogalactan protein epitopes to characterize water uptake pathways in leaves. Abaxially, the leaves of C. odoratissima are covered with peltate hairs, while the adaxial surfaces are glabrous. Both surfaces are able to absorb condensed water, but the lower surface has higher rates of water uptake. Numerous idioblasts connect the adaxial leaf surface and the abaxial peltate hairs, both of which contain hygroscopic substances such as arabinogalactan proteins and pectins. The highly specialized anatomy of the leaves of C odoratissima fulfills the dual function of minimizing water loss when stomata are closed, while maintaining the ability to absorb liquid water. Cell-wall related hygroscopic compounds in the peltate hairs and idioblasts create a network of microchannels that maintain leaf hydration and promote water uptake.

Heightened by the COVID-19 pandemic there has been a global increase in urban greenspace appreciation. Indoor plants are equally important for improving mental health and air quality but despite evolving in humid (sub)tropical environments with aerial root types, planting systems ignore aerial resource supply. This study directly compared nutrient uptake preferences of aerial and soil-formed roots of three common houseplant species under high and ambient relative humidities. Growth and physiology parameters were measured weekly for Anthurium andreanum, Epipremnum aureum and Philodendron scandens grown in custom made growth chambers. Both aerial and soil-formed roots were then fed mixtures of nitrate, ammonium and glycine, with one source labelled with 15N to determine uptake rates and maximum capacities. Aerial roots were consistently better at nitrogen uptake than soil roots but no species, root type or humidity condition showed a preference for a particular nitrogen source. All three species grew more in high humidity, with aerial roots demonstrating the greatest biomass increase. Higher humidities for indoor niches, together with fertiliser applications to aerial roots will support indoor plant growth, creating lush calming indoor environments for people inhabitants.

The 18O enrichment (Δ 18O) of leaf water affects the Δ 18O of photosynthetic products such as sucrose, generating an isotopic archive of plant function and past climate. However, uncertainty remains regarding how leaf water compartmentation between photosynthetic and non-photosynthetic tissue affects the relationship between Δ 18O of bulk leaf water (Δ 18O LW) and leaf sucrose (Δ 18O Sucrose). We grew Lolium perenne (a C 3 grass) in mesocosm-scale, replicated experiments with daytime relative humidity (RH 50 or 75%) and CO 2 level (200, 400 or 800 μmol mol -1) as factors, and determined Δ 18O LW, Δ 18O Sucrose and morpho-physiological leaf parameters, including transpiration ( E leaf), stomatal conductance ( g s) and mesophyll conductance to CO 2 ( g m). The Δ 18O of photosynthetic medium water (Δ 18O SSW) was estimated from Δ 18O Sucrose and the equilibrium fractionation between water and carbonyl groups (ε bio). Δ 18O SSW was well predicted by theoretical estimates of leaf water at the evaporative site (Δ 18O e) with adjustments that correlated with gas exchange parameters ( g s or total conductance to CO 2). Isotopic mass balance and published work indicated that non-photosynthetic tissue water was a large fraction (~0.53) of bulk leaf water. Δ 18O LW was a poor proxy for Δ 18O Sucrose, mainly due to opposite Δ 18O responses of non-photosynthetic tissue water (Δ 18O non-SSW) relative to Δ 18O SSW, driven by atmospheric conditions.