Plant Energy Metabolism Research Articles

Integration of Mitoflash and Time-Series Transcriptomics Facilitates Energy Dynamics Tracking and Substrate Supply Analysis of Floral Thermogenesis in Lotus.

The high biosynthetic and energetic demands of floral thermogenesis render thermogenic plants the ideal systems to characterize energy metabolism in plants, but real-time tracking of energy metabolism in plant cells remains challenging. In this study, a new method was developed for tracking the mitochondrial energy metabolism at the single mitochondria level by real-time imaging of mitochondrial superoxide production (i.e., mitoflash). Using this method, we observed the increased mitoflash frequencies in the receptacles of Nelumbo nucifera Gaertn. at the thermogenic stages. This increase, combined with the higher expression of antioxidant response-related genes identified through time-series transcriptomics at the same stages, shows us a new regulatory mechanism for plant redox balance. Furthermore, we found that the upregulation of respiratory metabolism-related genes during the thermogenic stages not only correlates with changes in mitoflash frequency but also underscores the critical roles of these pathways in ensuring adequate substrate supply for thermogenesis. Metabolite analysis revealed that sugars are likely one of the substrates for thermogenesis and may be transported over long distances by sugar transporters. Taken together, our findings demonstrate that mitoflash is a reliable tool for tracking energy metabolism in thermogenic plants and contributes to our understanding of the regulatory mechanisms underlying floral thermogenesis.

Identification of Rapeseed (Brassica napus L.) Plant Height-Associated QTL Using BSA-seq and RNA-seq.

Plant height (PH) is a critical agronomic trait in Brassica napus, significantly impacting yield. Consequently, identifying genes associated with plant height is a pivotal objective in oilseed rape breeding. This study employed a combination of bulk segregant analysis sequencing (BSA-seq) and RNA sequencing (RNA-seq) for analysis. A novel quantitative trait locus (QTL), qPH_C02, was identified between 63,989,634 and 64,945,122 bp on chromosome C02, from which eight candidate genes were screened. The Gene Ontology (GO) analysis revealed enrichment in peroxisomes, while the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated enrichment in the oxidative phosphorylation (OP) pathway. It is hypothesized that the observed differences in plant height and silique length may be attributed to the regulation of peroxidase activity in the OP pathway, which in turn alters plant energy metabolism and controls nutrient uptake. Subsequently, we will further test this hypothesis. The results of this study will contribute to our understanding of the genetic basis for differences in plant height and provide a foundation for the selection and breeding of Brassica napus varieties with desired plant shapes.

Genome-wide identification of starch phosphorylase gene family in Rosa chinensis and expression in response to abiotic stress

Chinese rose (Rosa chinensis) is an important ornamental plant, with economic, cultural, and symbolic significance. During the application of outdoor greening, adverse environments such as high temperature and drought are often encountered, which affect its application scope and ornamental quality. The starch phosphorylase (Pho) gene family participate in the synthesis and decomposition of starch, not only related to plant energy metabolism, but also plays an important role in plant stress resistance. The role of Pho in combating salinity and high temperature stress in R. chinensis remains unknown. In this work, 4 Phos from R. chinensis were detected with Pfam number of Pho (PF00343.23) and predicted by homolog-based prediction (HBP). The Phos are characterized by sequence lengths of 821 to 997 bp, and the proteins are predicted to subcellularly located in the plastid and cytoplasm. The regulatory regions of the Phos contain abundant stress and phytohormone-responsive cis-acting elements. Based on transcriptome analysis, the Phos were found to respond to abiotic stress factors such as drought, salinity, high temperature, and plant phytohormone of jasmonic acid and salicylic acid. The response of Phos to abiotic stress factors such as salinity and high temperature was confirmed by qRT-PCR analysis. To evaluate the genetic characteristics of Phos, a total of 69 Phos from 17 species were analyzed and then classified into 3 groups in phylogenetic tree. The collinearity analysis of Phos in R. chinensis and other species was conducted for the first time. This work provides a view of evolution for the Pho gene family and indicates that Phos play an important role in abiotic stress response of R. chinensis.

Selenium Improves the Control Efficacy of Phytophthora nicotianae by Damaging the Cell Membrane System and Promoting Plant Energy Metabolism.

Tobacco black shank (TBS), caused by Phytophthora nicotianae, poses a significant threat to tobacco plants. Selenium (Se), recognized as a beneficial trace element for plant growth, exhibited inhibitory effects on P. nicotianae proliferation, disrupting the cell membrane integrity. This action reduced the energy supply and hindered hyphal transport through membrane proteins, ultimately inducing hyphal apoptosis. Application of 8 mg/L Se through leaf spraying resulted in a notable decrease in TBS incidence. Moreover, Se treatment preserved chloroplast structure, elevated chitinase activities, β-1,3-GA, polyphenol oxidase, phenylalanine ammonia-lyase, and increased hormonal content. Furthermore, Se enhanced flavonoid and sugar alcohol metabolite levels while diminishing amino acid and organic acid content. This shift promoted amino acid degradation and flavonoid synthesis. These findings underscore the potential efficacy of Se in safeguarding tobacco and potentially other plants against P. nicotianae.

Ethylene inhibits photosynthesis via temporally distinct responses in tomato plants.

Ethylene is a volatile plant hormone that regulates many developmental processes and responses toward (a)biotic stress. Studies have shown that high levels of ethylene repress vegetative growth in many important crops, including tomato (Solanum lycopersicum), possibly by inhibiting photosynthesis. We investigated the temporal effects of ethylene on young tomato plants using an automated ethylene gassing system to monitor the physiological, biochemical, and molecular responses through time course RNA-seq of a photosynthetically active source leaf. We found that ethylene evokes a dose-dependent inhibition of photosynthesis, which can be characterized by 3 temporally distinct phases. The earliest ethylene responses that marked the first phase and occurred a few hours after the start of the treatment were leaf epinasty and a decline in stomatal conductance, which led to lower light perception and CO2 uptake, respectively, resulting in a rapid decline of soluble sugar levels (glucose, fructose). The second phase of the ethylene effect was marked by low carbohydrate availability, which modulated plant energy metabolism to adapt by using alternative substrates (lipids and proteins) to fuel the TCA cycle. Long-term continuous exposure to ethylene led to the third phase, characterized by starch and chlorophyll breakdown, which further inhibited photosynthesis, leading to premature leaf senescence. To reveal early (3 h) ethylene-dependent regulators of photosynthesis, we performed a ChIP-seq experiment using anti-ETHYLENE INSENSITIVE 3-like 1 (EIL1) antibodies and found several candidate transcriptional regulators. Collectively, our study revealed a temporal sequence of events that led to the inhibition of photosynthesis by ethylene and identified potential transcriptional regulators responsible for this regulation.

Fluorescein staining of chloroplast starch granules in living plants.

Chloroplast starch granules (cpSGs) store energy harvested through photosynthesis in plants, and cpSG dynamics have important roles in plant energy metabolism and stress responses. To date, cpSGs have been visualized using several methods, such as iodine staining; however, no method can be used to specifically visualize cpSGs in living cells from various plant species. Here, we report a simple method to visualize cpSGs in living plant cells in various species by staining with fluorescein, a commonly used fluorescent dye. We show that fluorescein is taken up into chloroplasts and interacts with cpSGs similarly to iodine. Fluorescein also interacts with refined starch in vitro. Using a fluorescein derivative for ultrabright cpSG imaging, we produced high-quality 3D reconstructions of cpSGs and evaluated their accumulation in multiple plant species. As fluorescein is well known and readily purchasable, our fluorescein-based staining method should contribute to all research regarding starch.

Genome-Wide Identification and Expression Profiling Analysis of SWEET Family Genes Involved in Fruit Development in Plum (Prunus salicina Lindl).

SWEETs (sugars will eventually be exported transporters) play a vital role in longer-distance sugar transportation, and thus control carbon flow and energy metabolism in plants. SWEET genes have been identified in various plant species, but their functions in fruit development remain uncharacterized. Here, we isolated 15 putative PsSWEETs from the Prunus salicina genome. For further analysis, comprehensive bioinformatics methods were applied to determine the gene structure, chromosome distribution, phylogeny, cis-acting regulatory elements, and expression profiles of PsSWEETs. qRT-PCR analysis suggested that these SWEETs might have diverse functions in the development of plum fruit. The relative expression levels of PsSWEET1 and PsSWEET9 were obviously higher in ripened fruit than the ones in other developmental stages, suggesting their possible roles in the transport and accumulation of sugars in plum fruit. Positive correlations were found between the expression level of PsSWEET3/10/13 and the content of sucrose, and the expression level of PsSWEET2 and the content of fructose, respectively, during the development of 'Furongli' fruit, suggesting their possible roles in the accumulation of sucrose and fructose. The current study investigated the initial genomic characterization and expression patterns of the SWEET gene family in plum, which could provide a foundation for the further understanding of the functional analysis of the SWEET gene family.

Plant photosynthetic responses under drought stress: Effects and management

AbstractBalanced photosynthesis is essential for improved plant survival and agricultural benefits in terms of biomass and yield. Photosynthesis is the hub of energy metabolism in plants; however, drought stress (DS) strongly perturbs photosynthetic efficiency due to biochemical and diffusive limitations that reduce key photosynthetic components and close stomata. This review describes photosynthetic responses, chloroplast retrograde signalling, and genetic imprints that curtail DS damage to photosynthetic machinery. While stomatal closure, disrupted photosynthetic systems, over‐reduced electron transport rates (ETR), partial hindrance of the Calvin cycle, and reduced pigment contents strongly affect the repertoire of photosynthetic processes under DS, chloroplast retrograde signalling also has a plausible role in preserving photosynthetic capacity. Progress in agronomic, genetic engineering approaches and isoprene regulation would help to rescue photosynthetic apparatus under DS.

Identification and molecular characterization of novel sucrose transporters in the hexaploid wheat (Triticum aestivum L.)

Common wheat (Triticum aestivum) is a major cereal crop grown and consumed globally. Recent advances in sequencing technology have facilitated the exploration of large and repetitive genomes. Plant sucrose transporter (SUT) genes are vital components of energy transport systems that play prominent roles in various plant functions, such as signaling and stress regulation. In this study, we identified and analyzed five novel sucrose transporter genes in wheat. The wheat sucrose transporter genes were divided into five clades based on their phylogenetic relationships. Synteny analysis revealed that synteny in the genome is highly conserved between wheat and rye, barley, and Brachypodium. Furthermore, the cis-element analysis indicated that sucrose transporter genes might be regulated by light and some phytohormone-related transcriptional factors. Overall, plant tissue-specific gene expression revealed enhanced expression of the transporter genes in the root and stem, whereas they were differentially expressed under abiotic stress treatments (cold, heat, NaCl, PEG-6000, and sucrose). These results indicate that each TaSUT gene may play a crucial role in stabilizing plants under stress by actively regulating the energy demands of cells. The findings of this study may provide a basis for further research on sucrose transporters and their significant roles in plant energy metabolism as well as in abiotic stress response, signaling, and regulation.

Advances in the Bioactivities of Phytochemical Saponins in the Prevention and Treatment of Atherosclerosis

Atherosclerosis (AS) is a chronic inflammatory disease characterized by hardening and narrowing of arteries. AS leads to a number of arteriosclerotic vascular diseases including cardiovascular diseases, cerebrovascular disease and peripheral artery disease, which pose a big threat to human health. Phytochemicals are a variety of intermediate or terminal low molecular weight secondary metabolites produced during plant energy metabolism. Phytochemicals from plant foods (vegetables, fruits, whole grains) and traditional herb plants have been shown to exhibit multiple bioactivities which are beneficial for prevention and treatment against AS. Many types of phytochemicals including polyphenols, saponins, carotenoids, terpenoids, organic sulfur compounds, phytoestrogens, phytic acids and plant sterols have already been identified, among which saponins are a family of glycosidic compounds consisting of a hydrophobic aglycone (sapogenin) linked to hydrophilic sugar moieties. In recent years, studies have shown that saponins exhibit a number of biological activities such as anti-inflammation, anti-oxidation, cholesterol-lowering, immunomodulation, anti-platelet aggregation, etc., which are helpful in the prevention and treatment of AS. This review aims to summarize the recent advances in the anti-atherosclerotic bioactivities of saponins such as ginsenoside, soyasaponin, astra-galoside, glycyrrhizin, gypenoside, dioscin, saikosaponin, etc.

Between Light and Shading: Morphological, Biochemical, and Metabolomics Insights Into the Influence of Blue Photoselective Shading on Vegetable Seedlings.

High nursery densities reduce the seedling quality due to the competition for light. High light intensity, shading, and blue light depletion activate morphophysiological and metabolomic responses in plants, resulting in size modification to gain an advantage over neighboring plants. Our research aimed to unravel the effects of light intensity and quality on nursery seedlings at the morphological and biochemical levels. To this aim, the effect of black shading and blue photoselective shading nets were investigated in terms of morphometric, ionomic, and untargeted metabolomics signatures in Cucurbita pepo L., Citrullus lanatus L., Solanum lycopersicum L., and Solanum melongena L. seedlings. Plant height, diameter, sturdiness index, leaf area, specific leaf area, shoot/root ratio, and mineral content (by ion chromatography-IC) were evaluated. In C. pepo L and C. lanatus L., the blue net reduced the shoot/root and chlorophyll a/b ratios and increased stem diameter and total chlorophyll content. The black net increased plant height, stem diameter, and sturdiness index in Solanum lycopersicum L. and Solanum melongena L. At the same time, unshading conditions reduced leaf area, specific leaf area, shoot/root ratio, and total chlorophyll content. The blue net improved the sturdiness index and quality of C. pepo L. and C. lanatus L. Such impact on morphological parameters induced by the different shading conditions was corroborated by a significant modulation at the metabolomics level. Untargeted metabolomic phytochemical signatures of the selected plants, and the subsequent multivariate analysis coupled to pathway analysis, allowed highlighting a broad and diverse biochemical modulation. Metabolomics revealed that both primary and secondary metabolism were largely affected by the different shading conditions, regardless of the species considered. A common pattern arose to point at the activation of plant energy metabolism and lipid biosynthesis, together with a generalized down accumulation of several secondary metabolites, particularly phenylpropanoids. Our findings indicate an intriguing scientific interest in the effects of selective shading and its application to other species and different phenological stages.

Compartment-specific energy requirements of photosynthetic carbon metabolism in Camelina sativa leaves

Main conclusionThe oxidative pentose phosphate pathway provides cytosolic NADPH yet reduces carbon and energy use efficiency. Repressing this pathway and introducing cytosolic NADPH-dependent malate dehydrogenase may increase crop yields by ≈5%.Detailed knowledge about plant energy metabolism may aid crop improvements. Using published estimates of flux through central carbon metabolism, we phenotype energy metabolism in illuminated Camelina sativa leaves (grown at 22 °C, 500 µmol photons m−2 s−1) and report several findings. First, the oxidative pentose phosphate pathway (OPPP) transfers 3.3% of the NADPH consumed in the Calvin–Benson cycle to the cytosol. NADPH supply proceeds at about 10% of the rate of net carbon assimilation. However, concomitantly respired CO2 accounts for 4.8% of total rubisco activity. Hence, 4.8% of the flux through the Calvin–Benson cycle and photorespiration is spent on supplying cytosolic NADPH, a significant investment. Associated energy requirements exceed the energy output of the OPPP. Thus, autotrophic carbon metabolism is not simply optimised for flux into carbon sinks but sacrifices carbon and energy use efficiency to support cytosolic energy metabolism. To reduce these costs, we suggest bioengineering plants with a repressed cytosolic OPPP, and an inserted cytosolic NADPH-dependent malate dehydrogenase tuned to compensate for the loss in OPPP activity (if required). Second, sucrose cycling is a minor investment in overall leaf energy metabolism but a significant investment in cytosolic energy metabolism. Third, leaf energy balancing strictly requires oxidative phosphorylation, cofactor export from chloroplasts, and peroxisomal NADH import. Fourth, mitochondria are energetically self-sufficient. Fifth, carbon metabolism has an ATP/NADPH demand ratio of 1.52 which is met if ≤ 21.7% of whole electron flux is cyclic. Sixth, electron transport has a photon use efficiency of ≥ 62%. Last, we discuss interactions between the OPPP and the cytosolic oxidation–reduction cycle in supplying leaf cytosolic NADPH.

The Responses of Leaf Litter Calcium, Magnesium, and Manganese Dynamics to Simulated Nitrogen Deposition and Reduced Precipitation Vary with Different Decomposition Stages

Litter decomposition is a vital link between material circulation and energy flow in forest ecosystems and is intensely affected by global change factors, such as increased nitrogen (N) deposition and altered precipitation regimes. As essential nutrients, calcium (Ca), magnesium (Mg), and manganese (Mn) play crucial roles in plant energy metabolism, photosynthesis, and membrane transport of plants, and the major source of these nutrients is litter decomposition. However, the dynamics of Ca, Mg, and Mn during decomposition have been largely ignored. Thus, to better understand Ca, Mg, and Mn dynamics during leaf litter decomposition in the scenario of increasing N deposition and decreasing precipitation, we carried out a two-year field litterbag experiment in a natural evergreen broad-leaved forest in the central area of the rainy area of Western China. Two levels of N deposition (ambient N deposition and 150 kg·N·ha−1·y−1) and precipitation reduction (no throughfall reduction and 10% throughfall reduction) were set, i.e., control (Ctr: without nitrogen deposition or throughfall reduction), N deposition (N, 150 kg·N·ha−1·y−1), throughfall reduction (T, 10% throughfall reduction), and N deposition and throughfall reduction (NT, 150 kg·N·ha−1·y−1 and 10% throughfall reduction). We found that leaf litter Ca concentration increased in the early decomposition stage and then decreased, while Mg and Mn concentrations generally decreased during the whole period of decomposition. The amount of Ca showed an accumulation pattern, while Mg and Mn generally showed a release pattern. N deposition and throughfall reduction affected the Ca, Mg, and Mn dynamics, varying with different decomposition stages; i.e., N deposition significantly affected the concentration and amount of Ca, regardless of the decomposition stages, while throughfall reduction significantly affected the Ca concentration in the whole and early decomposition stages. N deposition significantly affected the concentration and amount of Mg in the whole and early decomposition stages, while throughfall reduction had no significant effects. Throughfall reduction significantly affected the concentration and amount of Mn in the whole and late decomposition stages, while N deposition had no significant effects. Ca concentration generally showed a significant positive linear relationship with mass loss in the early decomposition stage; Mg concentration showed a significant positive linear relationship with mass loss in the Ctr and N treatments in the early and late decomposition stages; Mn generally showed a significant negative linear relationship with mass loss, regardless of the decomposition stage. Overall, the results suggest that Ca accumulation is more likely affected by N deposition, while Mg and Mn releases are more likely affected by N deposition combined with throughfall reduction, particularly in the early decomposition stage.

The effect of Fusarium culmorum inoculation and deoxynivalenol application on proteome response in wheat cultivars Sumai 3 and SW Kadrilj

Fusarium head blight (FHB) represents a damaging disease of cereal spikes caused by mycotoxin-producing Fusarium fungi revealing adverse effects on grain quality and yield. Chinese spring wheat (Triticum aestivum L.) cv. Sumai 3 represents a major source of FHB resistance thanks to Fhb1 QTL on 3BS chromosome conferring resistance to FHB and encoding a lectin-like protein. The aim of our study lied in a comparison of proteome response to Fusarium culmorum inoculation and mycotoxin deoxynivalenol (DON) application at 10 d after inoculation (dai) in spikes of resistant wheat cultivar Sumai 3 and susceptible wheat cultivar SW Kadrilj. Proteome analysis revealed profound impact of Fusarium inoculation and mycotoxin application on plant energy metabolism. Fusarium inoculation decreased photosynthesis and ATP biosynthesis and increased the level of stress-protective proteins (chaperones such as Hsc70, lectins). Genotype related differences observed at 10 dai indicated an active acclimation in Sumai 3 as indicated by increased content of some enzymes involved in phenolics biosynthesis (phenylalanine ammonia lyase PAL, BAHD acyltransferase), jasmonate biosynthesis (lipoxygenase LOX), and oligosaccharide biosynthesis (sucrose synthase SuSy, UDP-glucose uridylyltransferase) which are an important part of glycoproteins such as lectins and other pathogen-responsive biomolecules. The study thus provides data on plant acclimation to Fusarium infection which may underlie superior resistance of Sumai 3 cultivar.

Dynamics of Calcium, Magnesium, and Manganese During Litter Decomposition in Alpine Forest Aquatic and Terrestrial Ecosystems

Plant litter decomposition is one of the most important ecosystem processes influencing elemental biogeochemistry across terrestrial and aquatic environments. Calcium (Ca), magnesium (Mg), and manganese (Mn) are essential nutrients, which play a key role in plant energy metabolism, photosynthesis, and membrane transport of plants, and the major source of these nutrients is litter decomposition. Although litter decomposition has been subject to extensive research over the last several decades, the dynamics of Ca, Mg, and Mn in decomposing litter has been poorly assessed. Also, very few decomposition studies have been simultaneously carried out across aquatic and terrestrial ecosystems. Here, we conducted a two-year litterbag experiment across alpine forest aquatic (stream and riparian zone) and terrestrial (forest floor) ecosystems to address the dynamics of Ca, Mg, and Mn during litter decomposition. We found that (1) in general concentrations of Ca, Mg, and Mn all increased (that is, high immobilization) during the period of decomposition; (2) Ca and Mn immobilized in decomposing litter regardless of ecosystem type or litter species identity, while Mg showed release (mineralization) or immobilization depending on litter species identity; (3) a common hierarchy of drivers for Ca, Mg, or Mn release rate was found across all ecosystem types; and (4) Ca, Mg, and Mn concentrations all showed positive relationships with litter decomposition. Our results provide evidence of general patterns of Ca, Mg, and Mn dynamics in decomposing litter and significant positive effects of these nutrients on litter decomposition across terrestrial and aquatic environments associated with a forest ecosystem. We suggest that addressing nutrient dynamics across contrasting environments is crucial for predicting how biogeochemical cycling of different nutrients is modulated by litter mass decomposition.

Mechanism study of the beneficial effect of sodium selenite on metabolic disorders in imidacloprid-treated garlic plants.

As an effective neonicotinoid insecticide, imidacloprid (IMI) has been widely used in crop production, but its residue affects normal plant growth. Selenium (Se) is a non-essential mineral nutrient in higher plants, that acts as the active centre of glutathione peroxidase (GSH-Px), which removes harmful peroxides. In this study, we investigated the mechanism by which selenium improves the growth status of IMI-treated garlic plants through analyses of apparent morphology and antioxidant enzyme activity as well as the dynamic changes in nutrients and metabolites in the plants. The results showed that 80μg/kg Na2SeO3 had a strong effect on alleviating the damage in garlic plants exposed to IMI (1.2mg/kg) by increasing the absorption of mineral elements to enhance the synthesis of chlorophyll and antioxidant enzymes. A nontarget metabolomics analysis based on gas chromatography-mass spectrometry (GC-MS) indicated that the addition of Na2SeO3 to IMI-treated garlic could reconstruct the plant metabolic distribution by enhancing the nitrogen and indole metabolism, maintaining lower concentrations of secondary metabolites and maintaining the balance of the plant energy metabolism. Our study provides novel insights into the molecular mechanisms by which garlic plants responds to IMI exposure and suggests the use of selenium with IMI-contaminated plants as a solution for the advancement of sustainable agricultural pesticide use.

Composition and Stability of the Oxidative Phosphorylation System in the Halophile Plant Cakile maritima.

Mitochondria play a central role in the energy metabolism of plants. At the same time, they provide energy for plant stress responses. We here report a first view on the mitochondrial Oxidative Phosphorylation (OXPHOS) system of the halophile (salt tolerant) plant Cakile maritima. Mitochondria were purified from suspension cultures of C. maritima and for comparison of Arabidopsis thaliana, a closely related glycophyte (salt sensitive) plant. Mitochondria were treated with digitonin and solubilized protein complexes were analyzed by 2D Blue native/SDS polyacrylamide gel electrophoresis. The OXPHOS systems of the two compared plants exhibit some distinct differences. C. maritima mitochondria include a very abundant respiratory supercomplex composed of monomeric complex I and dimeric complex III. At the same time the complexes II and IV are of reduced abundance. The stability of the OXPHOS complexes was investigated by combined salt and temperature treatments of isolated mitochondria. ATP synthase (complex V) is of increased stability in C. maritima. Also, the I + III2 supercomplex is present in high abundance during stress treatments. These results give insights into the mitochondrial contribution to the plant salt stress response.

2 H-fractionations during the biosynthesis of carbohydrates and lipids imprint a metabolic signal on the δ2 H values of plant organic compounds.

Hydrogen (H) isotope ratio (δ2 H) analyses of plant organic compounds have been applied to assess ecohydrological processes in the environment despite a large part of the δ2 H variability observed in plant compounds not being fully elucidated. We present a conceptual biochemical model based on empirical H isotope data that we generated in two complementary experiments that clarifies a large part of the unexplained variability in the δ2 H values of plant organic compounds. The experiments demonstrate that information recorded in the δ2 H values of plant organic compounds goes beyond hydrological signals and can also contain important information on the carbon and energy metabolism of plants. Our model explains where 2 H-fractionations occur in the biosynthesis of plant organic compounds and how these 2 H-fractionations are tightly coupled to a plant's carbon and energy metabolism. Our model also provides a mechanistic basis to introduce H isotopes in plant organic compounds as a new metabolic proxy for the carbon and energy metabolism of plants and ecosystems. Such a new metabolic proxy has the potential to be applied in a broad range of disciplines, including plant and ecosystem physiology, biogeochemistry and palaeoecology.

Sample Preparation for Analysis of the Plant Mitochondrial Membrane Proteome.

Containing plastids and vacuoles in addition to those organelles also found in other (heterotrophic) cells, the plant cell displays an extraordinary level of compartmentalization, largely obtained by the utilization of membranes. These membranes not only confine reaction spaces but must also facilitate cross-talk between organelles and other cell compartments. They also host important components of the plant energy metabolism, i.e., the electron transport chains of mitochondria and chloroplasts. Characterization of the proteomes of these membranes requires isolation of pure and intact organelles from plant tissues followed by subsequent purification of their respective membranes. Membrane fractions are then amenable for further analyses using gel electrophoresis procedures or gel-free proteomic approaches. Here, we describe the preparation of intact mitochondria from Arabidopsis thaliana cell-culture, the isolation of outer and inner mitochondrial membranes and downstream proteomic applications for analyzing their membrane protein content.

ER-localized adenine nucleotide transporter ER-ANT1: an integrator of energy and stress signaling in rice.

Most environmental perturbations have a direct or indirect deleterious impact on photosynthesis, and, in consequence, the overall energy status of the cell. Despite our increased understanding of convergent energy and stress signals, the connections between photosynthesis, energy and stress signals through putative common nodes are still unclear. Here we identified an endoplasmic reticulum (ER)-localized adenine nucleotide transporter1 (ER-ANT1), whose deficiency causes seedling lethality in air but viable under high CO2, exhibiting the typical photorespiratory phenotype. Metabolic analysis suggested that depletion of ER-ANT1 resulted in circadian rhythm disorders in sucrose synthesis and induced sucrose signaling pathways, indicating that the ER is involved in the regulation of vital energy metabolism in plants. In addition, the defect of ER-ANT1 triggers ER stress and activates the unfolded protein response in plant cells, suggesting ER stress and photorespiration are closely linked. These findings provide an important evidence for a key role of ER-localized ER-ANT1 in convergent energy and stress signals in rice. Our findings support the idea that ATP is a central signal involved in the plant response to a variety of stresses.