Alternative Respiratory Pathway Research Articles

Noncoupled Mitochondrial Respiration as Therapeutic Approach for the Treatment of Metabolic Diseases: Focus on Transgenic Animal Models.

Mitochondrial dysfunction contributes to numerous chronic diseases, and mitochondria are targets for various toxins and xenobiotics. Therefore, the development of drugs or therapeutic strategies targeting mitochondria is an important task in modern medicine. It is well known that the primary, although not the sole, function of mitochondria is ATP generation, which is achieved by coupled respiration. However, a high membrane potential can lead to uncontrolled reactive oxygen species (ROS) production and associated dysfunction. For over 50 years, scientists have been studying various synthetic uncouplers, and for more than 30 years, uncoupling proteins that are responsible for uncoupled respiration in mitochondria. Additionally, the proteins of the mitochondrial alternative respiratory pathway exist in plant mitochondria, allowing noncoupled respiration, in which electron flow is not associated with membrane potential formation. Over the past two decades, advances in genetic engineering have facilitated the creation of various cellular and animal models that simulate the effects of uncoupled and noncoupled respiration in different tissues under various disease conditions. In this review, we summarize and discuss the findings obtained from these transgenic models. We focus on the advantages and limitations of transgenic organisms, the observed physiological and biochemical changes, and the therapeutic potential of uncoupled and noncoupled respiration.

Germination of Pisum sativum L. Seeds Is Associated with the Alternative Respiratory Pathway.

The alternative oxidase (AOX) is a ubiquinol oxidase with a crucial role in the mitochondrial alternative respiratory pathway, which is associated with various processes in plants. In this study, the activity of AOX in pea seed germination was determined in two pea cultivars, 'Maravilha d'América' (MA) and 'Torta de Quebrar' (TQ), during a germination trial using cytochrome oxidase (COX) and AOX inhibitors [rotenone (RT) and salicylic hydroxamic acid (SHAM), respectively]. Calorespirometry was used to assess respiratory changes during germination. In both cultivars, SHAM had a greater inhibitory effect on germination than RT, demonstrating the involvement of AOX in germination. Although calorespirometry did not provide direct information on the involvement of the alternative pathway in seed germination, this methodology was valuable for distinguishing cultivars. To gain deeper insights into the role of AOX in seed germination, the AOX gene family was characterized, and the gene expression pattern was evaluated. Three PsAOX members were identified-PsAOX1, PsAOX2a and PsAOX2b-and their expression revealed a marked genotype effect. This study emphasizes the importance of AOX in seed germination, contributing to the understanding of the role of the alternative respiratory pathway in plants.

Antifungal Activity of 2-Allylphenol Derivatives on the Botrytis cinerea Strain: Assessment of Possible Action Mechanism.

Botrytis cinerea is a phytopathogenic fungus that causes serious damage to the agricultural industry by infecting various important crops. 2-allylphenol has been used in China as a fungicide for more than a decade, and it has been shown that is a respiration inhibitor. A series of derivatives of 2-allylphenol were synthesized and their activity against B. cinerea was evaluated by measuring mycelial growth inhibition. Results indicate that small changes in the chemical structure or the addition of substituent groups in the aromatic ring induce important variations in activity. For example, changing the hydroxyl group by methoxy or acetyl groups produces dramatic increases in mycelial growth inhibition, i.e., the IC50 value of 2-allylphenol decreases from 68 to 2 and 1 μg mL-1. In addition, it was found that the most active derivatives induce the inhibition of Bcaox expression in the early stages of B. cinerea conidia germination. This gene is associated with the activation of the alternative oxidase enzyme (AOX), which allows fungus respiration to continue in the presence of respiratory inhibitors. Thus, it seems that 2-allylphenol derivatives can inhibit the normal and alternative respiratory pathway of B. cinerea. Therefore, we believe that these compounds are a very attractive platform for the development of antifungal agents against B. cinerea.

Hydrogen sulfide upregulates the alternative respiratory pathway in mangrove plant Avicennia marina to attenuate waterlogging-induced oxidative stress and mitochondrial damage in a calcium-dependent manner.

Hydrogen sulfide (H2 S) is considered to mediate plant growth and development. However, whether H2 S regulates the adaptation of mangrove plant to intertidal flooding habitats is not well understood. In this study, sodium hydrosulfide (NaHS) was used as an H2 S donor to investigate the effect of H2 S on the responses of mangrove plant Avicennia marina to waterlogging. The results showed that 24-h waterlogging increased reactive oxygen species (ROS) and cell death in roots. Excessive mitochondrial ROS accumulation is highly oxidative and leads to mitochondrial structural and functional damage. However, the application of NaHS counteracted the oxidative damage caused by waterlogging. The mitochondrial ROS production was reduced by H2 S through increasing the expressions of the alternative oxidase genes and increasing the proportion of alternative respiratory pathway in the total mitochondrial respiration. Secondly, H2 S enhanced the capacity of the antioxidant system. Meanwhile, H2 S induced Ca2+ influx and activated the expression of intracellular Ca2+ -sensing-related genes. In addition, the alleviating effect of H2 S on waterlogging can be reversed by Ca2+ chelator and Ca2+ channel blockers. In conclusion, this study provides the first evidence to explain the role of H2 S in waterlogging adaptation in mangrove plants from the mitochondrial aspect.

Mitochondrial alternative oxidase enhanced ABA-mediated drought tolerance in Solanum lycopersicum

The phytohormone abscisic acid (ABA) plays essential roles in modulating drought stress responses. Mitochondrial alternative oxidase (AOX) is critical for reactive oxygen species (ROS) scavenging in drought stress responses. However, whether ABA signal in concert with AOX to moderate drought stress response remains largely unclear. In our study, we uncover the positive role of AOX in ABA-mediated drought tolerance in tomato (Solanum lycopersicum). Here, we report that ABA participates in the regulation of alternative respiration, and the increased AOX was found to improve drought tolerance by reducing total ROS accumulation. We also found that transcription factor ABA response element-binding factor 1 (SlAREB1) can directly bind to the promoter of AOX1a to activate its transcription. Virus-induced gene silencing (VIGS) of SlAREB1 compromised the ABA-induced alternative respiratory pathway, disrupted redox homeostasis and decreased plant resistance to drought stress, while overexpression of AOX1a in TRV2-SlAREB1 plants partially rescued the severe drought phenotype. Taken together, our results indicated that AOX1a plays an essential role in ABA-mediated drought tolerance partially in a SlAREB1-dependent manner, providing new insights into how ABA modulates ROS levels to cope with drought stress by AOX.

Ascorbate synthesis as an alternative electron source for mitochondrial respiration: Possible implications for the plant performance.

The molecule vitamin C, in the chemical form of ascorbic acid (AsA), is known to be essential for the metabolism of humans and animals. Humans do not produce AsA, so they depend on plants as a source of vitamin C for their food. The AsA synthesis pathway occurs partially in the cytosol, but the last oxidation step is physically linked to the respiratory chain of plant mitochondria. This oxidation step is catalyzed by l-galactono-1,4-lactone dehydrogenase (l-GalLDH). This enzyme is not considered a limiting step for AsA production; however, it presents a distinguishing characteristic: the l-GalLDH can introduce electrons directly into the respiratory chain through cytochrome c (Cytc) and therefore can be considered an extramitochondrial electron source that bypasses the phosphorylating Complex III. The use of Cytc as electron acceptor has been debated in terms of its need for AsA synthesis, but little has been said in relation to its impact on the functioning of the respiratory chain. This work seeks to offer a new view about the possible changes that result of the link between AsA synthesis and the mitochondrial respiration. We hypothesized that some physiological alterations related to low AsA may be not only explained by the deficiency of this molecule but also by the changes in the respiratory function. We discussed some findings showing that respiratory mutants contained changes in AsA synthesis. Besides, recent works that also indicate that the excessive electron transport via l-GalLDH enzyme may affect other respiratory pathways. We proposed that Cytc reduction by l-GalLDH may be part of an alternative respiratory pathway that is active during AsA synthesis. Also, it is proposed that possible links of this pathway with other pathways of alternative electron transport in plant mitochondria may exist. The review suggests potential implications of this relationship, particularly for situations of stress. We hypothesized that this pathway of alternative electron input would serve as a strategy for adaptation of plant respiration to changing conditions.

Photosynthesis, Respiration, and Thermal Energy Dissipation in Leaves of Two Phenotypes of Plantago media L. under Environmental Conditions

The ability to maintain the balance between the absorbed light energy and energy used in photosynthesis is a key factor of plant adaptation to variable environmental conditions. In this work, diurnal variations in photosynthesis, respiration, thermal energy dissipation, and the activity of the antioxidant system were studied in hoary plantain (Plantago media L.) growing on an open slope (sun plants) and under natural shading in the herbage (shade plants). The highest leaves net photosynthetic rate (Pn) was observed early in the morning and amounted to 2.6 and 9.2 μmol CO2/m2 s in shade and sun plants, respectively. In the daytime, the Pn values of sun plants decreased significantly (threefold) along with the decrease in stomatal conductance; changes of both parameters developed concurrently with the increase in insolation and air temperature. The Pn changes in leaves of shade plants were less pronounced and weakly dependent on stomatal conductance. The leaves of shade plants contained comparatively high amounts of soluble carbohydrates, whereas the sun plant leaves accumulated larger amounts of starch. In the daytime, nonphotochemical quenching (NPQ) of chlorophyll a fluorescence in photosystem II of sun plant leaves could be as large as 2.6 rel. units, which was four- to fivefold higher than NPQ in shade plants. In leaves of sun plants in the morning and evening hours, the ratio of cytochrome pathway (CP) and the alternative (AP) respiratory pathways was approximately 1.0, whereas this ratio decreased to 0.4 during the day, synchronously with an increase in NPQ. The CP/AP ratio in shade plant leaves remained constant throughout the diurnal cycle and equaled 1.4, indicating a comparatively high energy efficiency of respiration in shaded plants growing under the grass canopy. The leaves of sun plants featured an increased content of superoxide anion radical and hydrogen peroxide as well as the elevated activity of antioxidant enzymes (superoxide dismutase, guaiacol peroxidase) that control the accumulation of reactive oxygen species. The results emphasize the importance of coordinated changes in energy-dissipating processes and the activity of the antioxidant system for maintaining the energy and redox balance in phototrophic tissues during long-term adaptation of plants to environmental conditions, excessive insolation in particular.

Direct electrochemical detection of trans-plasma membrane electron transfer: A possible alternative pathway for cell respiration

An electrochemical protocol was designed to enable Vero cells to transfer electrons to an electrode without any added redox mediator. The cells were cultured on the surface of electrodes polarized at the optimal potential of 400 mV/silver pseudo-reference. Gold, carbon, and CNT-coated carbon electrodes displayed similar current record patterns. Extracellular electron transfer was sustained for several days. Its intensity, up to 1.5 pA.cell−1, was in the range of the electron flows implemented by cell respiration. A large fraction of the current vanished as soon as anoxic conditions were established, which suggests a mitochondrial origin for a large proportion of the electrons. The current records always showed a two-phase pattern. The occurrence of the two phases was not due to an evolution of the cell mat structure, which was fully established during the first day of polarization and did not change significantly thereafter. Increasing the cell seeding density decreased the maximum current reached during the first phase and the duration of the phase. These observations put together lead us to propose a model, in which only the cells adhered on the electrode surface produced current by metabolizing glutamine during the first phase. The possible role of this extracellular electron transfer as an alternative cell respiration pathway is discussed. The key roles it could play in regulating pH and pO2 gradients are considered, specifically to explain the pH gradient reversal observed in cancer cells. These pioneering results pave the way for electrochemical sensors to directly address cellular metabolic pathways.

Pentatricopeptide repeat protein MITOCHONDRIAL STABILITY FACTOR 3 ensures mitochondrial RNA stability and embryogenesis.

Gene expression in plant mitochondria is predominantly governed at the post-transcriptional level and relies mostly on nuclear-encoded proteins. However, the protein factors involved and the underlying molecular mechanisms are still not well understood. Here, we report on the function of the MITOCHONDRIAL STABILITY FACTOR 3 (MTSF3) protein, previously named EMBRYO DEFECTIVE 2794 (EMB2794), and show that it is essential for accumulation of the mitochondrial NADH dehydrogenase subunit 2 (nad2) transcript in Arabidopsis (Arabidopsis thaliana) but not for splicing of nad2 intron 2 as previously proposed. The MTSF3 gene encodes a pentatricopeptide repeat protein that localizes in the mitochondrion. An MTSF3 null mutation induces embryonic lethality, but viable mtsf3 mutant plants can be generated through partial complementation with the developmentally regulated ABSCISIC ACID INSENSITIVE3 promoter. Genetic analyses revealed growth retardation in rescued mtsf3 plants owing to the specific destabilization of mature nad2 mRNA and a nad2 precursor transcript bearing exons 3 to 5. Biochemical data demonstrate that MTSF3 protein specifically binds to the 3' terminus of nad2. Destabilization of nad2 mRNA induces a substantial decrease in complex I assembly and activity and overexpression of the alternative respiratory pathway. Our results support a role for MTSF3 protein in protecting two nad2 transcripts from degradation by mitochondrial exoribonucleases by binding to their 3' extremities.

Impairment of Respiratory Chain Function and Involvement of Alternative Respiratory Pathway in Mitochondria of Potato Tubers Infected by Pectobacteriumcarotovorum subsp. carotovorum.

The significance of alternative respiratory pathway (AOXs) during the interaction between soft rot bacteria (Pectobacterium carotovorum subsp. carotovorum, (Pcc.)) and potato tubers is well-defined. However, the role of the AOXs in impaired mitochondrial respiratory chain function during the Pcc. infection is yet to be studied. In this study, the results show that with the aggravation of infection of Pcc., the capacity for alternative respiration in mitochondria of potato tubers increased gradually. The mitochondrial membrane potential increased more significantly after infection with Pcc. when the AOXs in potato tubers was partially blocked using salicylhydroxamic acid (SHAM) beforehand. In addition, the activity of complex III decreased more drastically while the activity of complex IV increased more significantly in the partial absence of the AOXs in the mitochondria. Furthermore, the mitochondrial endogenous respiration, mitochondrial respiratory state 3 and respiratory control rate (RCR) decreased more significantly and the value of RCR reached around 1.0 with the aggravation of infection of Pcc. in the partially absence of AOXs in the mitochondria.

Carbon metabolic adjustment in soybean nodules in response to phosphate limitation: A metabolite perspective

The necessity to gain deep insights regarding the overall metabolic efficiency of soybean functional acclimation to tackle phosphate (Pi) starvation has led us to discuss the current knowledge on the carbon (C) metabolic changes in soybean nodules under such Pi stress conditions in this review. Soybean possesses numerous coping strategies to conserve Pi utilization, while decreasing the C cost to promote nodule function during Pi stress. This could be achieved through the modulation of carbohydrate importation, mobilization of storage reserves, modification of respiratory pathways and exportation of nitrogenous (N) products. Under such circumstances, soybean regulates the C partitioning among the various nutrient-acquiring structures and the C flux through different metabolic pathways (primary and secondary). However, substantial genetic variation leading to nodule acclimation to Pi stress is notable and dependent on the crop genotypes and/or rhizobial strains that are in symbiosis. Several Pi deficiency-induced responses (e.g., mycorrhizal association, exudative burst and secondary metabolism) can act to increase the Pi levels in nodules, which are, unavoidably, often associated with a heavy burden to the overall C budget of the host. Alternative non-adenylate respiratory pathways (e.g., glycolytic bypass) and ureide export (i.e., less amide transport) are metabolically less expensive, and thus more favorable during Pi stress. • Pi starvation affects carbon (C) metabolic activities in soybean nodules. • Soybean possesses numerous coping strategies to optimize nodular C cost. • Rhizobial microsymbionts markedly affect the Pi deficiency-induced C expenditure. • C acclimation to Pi stress involves primary and secondary metabolic pathways. • Non-adenylate respiratory pathways and ureide export are metabolically less expensive.

The Melatonin Receptor CAND2/PMTR1 Is Involved in the Regulation of Mitochondrial Gene Expression under Photooxidative Stress.

Melatonin is a signaling molecule that mediates multiple stress-dependent reactions. Under photooxidative stress conditions generating intensive ROS production, exogenous melatonin (50 µM) contributed to maintaining the expression of mitochondrial encoded genes and up-regulation of RNA-polymerase genes RPOTm and RPOTmp, operating through the CAND2 receptor and α-subunit of the heterotrimeric G protein GPA1 coupled with CAND2. Unlike wild-type plants, mutants with defective CAND2 and GPA1 genes exhibited no decrease in the alternative pathway of leaf respiration, as well as the activity of an alternative oxidase, and the expression of the AOX1a gene. At the same time, the protective effect of exogenous melatonin on some physiological indicators did not depend on the receptor and was associated with the direct antioxidant function of the regulator. Thus, melatonin under photooxidative stress conditions can act as an antioxidant and as a hormone capable of regulating the expression of nuclear and organelle genes through the components of melatonin signal perception.

Alternative oxidase is involved in oxidative stress resistance and melanin synthesis in Annulohypoxylon stygium, a companion fungus of Tremella fuciformis.

Annulohypoxylon stygium, a companion fungus of Tremella fuciformis, provides nutrition for growth and development of T. fuciformis. Alternative oxidase (AOX), which is the terminal oxidase of the alternative respiration pathway, functions to transfer electrons from ubiquinol to O2 with the production of H2O. In this study, an AOX encoded gene, Asaox, was cloned from A. stygium. The coding sequence of Asaox gene contains 1056 nucleotides and encodes 351 amino acids. RNA interference was used to study the function of Asaox in A. stygium. The Asaox-silenced strains were confirmed by PCR, quantitative real-time PCR, and Southern blot. The Asaox-silenced strains exhibited increased relative growth inhibition rates when inoculated on PDA plates with 50mM H2O2, compared with wild-type strain. The growth rate in sawdust medium, melanin content, and laccase activity of Asaox-silenced strains were decreased. The expression levels of tyr and pks genes were decreased in Asaox-silenced strains, indicating that there might be two melanin synthesis pathways, DHN and DOPA, in A. stygium. In summary, the results have demonstrated that Asaox gene was involved in oxidative stress resistance and melanin synthesis in A. stygium.

The Arabidopsis electron-transfer flavoprotein:ubiquinone oxidoreductase is required during normal seed development and germination.

The importance of the alternative donation of electrons to the ubiquinol pool via the electron-transfer flavoprotein/electron-transfer flavoprotein:ubiquinone oxidoreductase (ETF/ETFQO) complex has been demonstrated. However, the functional significance of this pathway during seed development and germination remains to be elucidated. To assess the function of this pathway, we performed a detailed metabolic and transcriptomic analysis of Arabidopsis mutants to test the molecular consequences of a dysfunctional ETF/ETFQO pathway. We demonstrate that the disruption of this pathway compromises seed germination in the absence of an external carbon source and also impacts seed size and yield. Total protein and storage protein content is reduced in dry seeds, whilst sucrose levels remain invariant. Seeds of ETFQO and related mutants were also characterized by an altered fatty acid composition. During seed development, lower levels of fatty acids and proteins accumulated in the etfqo-1 mutant as well as in mutants in the alternative electron donors isovaleryl-CoA dehydrogenase (ivdh-1) and d-2-hydroxyglutarate dehydrogenase (d2hgdh1-2). Furthermore, the content of several amino acids was increased in etfqo-1 mutants during seed development, indicating that these mutants are not using such amino acids as alternative energy source for respiration. Transcriptome analysis revealed alterations in the expression levels of several genes involved in energy and hormonal metabolism. Our findings demonstrated that the alternative pathway of respiration mediated by the ETF/ETFQO complex affects seed germination and development by directly adjusting carbon storage during seed filling. These results indicate a role for the pathway in the normal plant life cycle to complement its previously defined roles in the response to abiotic stress.

CFM6 is an Essential CRM Protein Required for the Splicing of nad5 Transcript in Arabidopsis Mitochondria.

Plant chloroplast RNA splicing and ribosome maturation (CRM)-domain-containing proteins are capable of binding RNA to facilitate the splicing of group I or II introns in chloroplasts, but their functions in mitochondria are less clear. In the present study, Arabidopsis thaliana CFM6, a protein with a single CRM domain, was expressed in most plant tissues, particularly in flower tissues, and restricted to mitochondria. Mutation of CFM6 causes severe growth defects, including stunted growth, curled leaves, delayed embryogenesis and pollen development. CFM6 functions specifically in the splicing of group II intron 4 of nad5, which encodes a subunit of mitochondrial complex I, as evidenced by the loss of nad5 intron 4 splicing and high accumulation of its pretranscripts in cfm6 mutants. The phenotypic and splicing defects of cfm6 were rescued in transgenic plants overexpressing 35S::CFM6-YFP. Splicing failure in cfm6 also led to the loss of complex I activity and to its improper assembly. Moreover, dysfunction of complex I induced the expression of proteins or genes involved in alternative respiratory pathways in cfm6. Collectively, CFM6, a previously uncharacterized CRM domain-containing protein, is specifically involved in the cis-splicing of nad5 intron 4 and plays a pivotal role in mitochondrial complex I biogenesis and normal plant growth.

A strategy for promoting carbon flux into fatty acid and astaxanthin biosynthesis by inhibiting the alternative oxidase respiratory pathway in Haematococcus pluvialis

In the present study, the mechanisms for facilitating fatty acid and astaxanthin biosynthesis-related processes by inhibiting the alternative oxidase (AOX) respiratory pathway in Haematococcus pluvialis was investigated. The restriction of the AOX pathway induced the accumulation of reactive oxygen species, NAD(P)H and its substrates (acetyl-CoA, pyruvate and glyceraldehy-3-phosphate), which are required for fatty acid and astaxanthin production, thereby promoting the carbon flux into fatty acid and astaxanthin biosynthesis. During a 9-day incubation period, the fatty acid and astaxanthin contents increased by 20.6% and 20.7%, respectively, when the AOX pathway was inhibited approximately 37.7%. The AOX pathway may be inhibited by nutrient (nitrogen and phosphorus) removal, inhibitor addition and air/CO2 aeration adjustments in the large-scale cultivation of H. pluvialis. Therefore, the current study provides a useful enhancement strategy for fatty acid and astaxanthin coproduction and elucidates the roles of the AOX pathway in regulating fatty acid and astaxanthin biosynthesis.

Respiration responses of wheat seedlings to treatment with trehalose under heat stress

Heat stress limits wheat production and trehalose can improve stress tolerance. How trehalose affects wheat respiration is unclear. In this study, we investigated the effects of exogenous trehalose on the respiration of wheat seedlings during heat stress and the subsequent recovery period. Trehalose pretreatment significantly increased the expression of the alternative oxidase genes AOX1a and AOX1c under heat stress, indicating that trehalose pretreatment increased the capacity of the alternative respiration pathway (AP) in response to heat stress. Trehalose pretreatment also enhanced the activity of the malate-oxaloacetate (Mal-OAA) shuttle and ameliorated the decrease in photosynthetic activity caused by heat stress. However, when the AP was inhibited by salicylhydroxamic acid under heat stress, both Mal-OAA shuttle activity and photosynthetic efficiency were substantially reduced in the control and trehalose pretreatment groups. In addition, trehalose pretreatment helped to maintain inner mitochondrial respiratory activity and the activity of Complex II during heat stress, particularly the coupling of oxidative phosphorylation with the Complex II electron transport chain, thereby mitigating heat-related damage to the cytochrome pathway (CP). Taken together, these results suggest that exogenous trehalose enhanced the AP and reduced damage to the CP under heat stress in wheat seedlings, thus maintaining cellular energy metabolism. Up-regulation of the AP by trehalose pretreatment may improve the heat tolerance of wheat seedlings by dissipating excess reducing equivalents transported through the Mal-OAA shuttle, thereby protecting photosynthetic performance.

Distribution of Denitrification among Haloarchaea: A Comprehensive Study.

Microorganisms from the Halobacteria class, also known as haloarchaea, inhabit a wide range of ecosystems of which the main characteristic is the presence of high salt concentration. These environments together with their microbial communities are not well characterized, but some of the common features that they share are high sun radiation and low availability of oxygen. To overcome these stressful conditions, and more particularly to deal with oxygen limitation, some microorganisms drive alternative respiratory pathways such as denitrification. In this paper, denitrification in haloarchaea has been studied from a phylogenetic point of view. It has been demonstrated that the presence of denitrification enzymes is a quite common characteristic in Halobacteria class, being nitrite reductase and nitric oxide reductase the enzymes with higher co-occurrence, maybe due to their possible role not only in denitrification, but also in detoxification. Moreover, copper-nitrite reductase (NirK) is the only class of respiratory nitrite reductase detected in these microorganisms up to date. The distribution of this alternative respiratory pathway and their enzymes among the families of haloarchaea has also been discussed and related with the environment in which they constitute the major populations. Complete denitrification phenotype is more common in some families like Haloarculaceae and Haloferacaceae, whilst less common in families such as Natrialbaceae and Halorubraceae.

OsNBL3, a mitochondrion-localized pentatricopeptide repeat protein, is involved in splicing nad5 intron 4 and its disruption causes lesion mimic phenotype with enhanced resistance to biotic and abiotic stresses.

SummaryLesion mimic mutants are used to elucidate mechanisms controlling plant responses to pathogen attacks and environmental stresses. Although dozens of genes had been functionally demonstrated to be involved in lesion mimic phenotype in several plant species, the molecular mechanisms underlying the hypersensitive response are largely unknown. Here, a rice (Oryza sativa) lesion mimic mutant natural blight leaf 3 (nbl3) was identified from T‐DNA insertion lines. The causative gene, OsNBL3, encodes a mitochondrion‐localized pentatricopeptide repeat (PPR) protein. The nbl3 mutant exhibited spontaneous cell death response and H2O2 accumulation, and displayed enhanced resistance to the fungal and bacterial pathogens Magnaporthe oryzae and Xanthomonas oryzae pv. oryzae. This resistance was consistent with the up‐regulation of several defence‐related genes; thus, defence responses were induced in nbl3. RNA interference lines of OsNBL3 exhibited enhanced disease resistance similar to that of nbl3, while the disease resistance in overexpression lines did not differ from that of the wild type. In addition, nbl3 displayed improved tolerance to salt, accompanied by up‐regulation of several salt‐associated marker genes. OsNBL3 was found to mainly participate in the splicing of mitochondrial gene nad5 intron 4. Disruption of OsNBL3 leads to the reduction in complex I activity, the elevation of alternative respiratory pathways and the destruction of mitochondrial morphology. Overall, the results demonstrated that the PPR protein‐coding gene OsNBL3 is essential for mitochondrial development and functions, and its disruption causes the lesion mimic phenotype and enhances disease resistance and tolerance to salt in rice.

Effect of Shungite Application on the Temperature Sensitivity of Allium cepa Respiration under Two Soil Water Regimes

For agricultural soils with low natural fertility, the possibility of using rock powders as an alternative source of nutrients and/or improver of soil physical parameters is under discussion and study. Shungite rocks, carbon-bearing volcanic sedimentary rock, are characterized by a high content of carbon and nutrients. This study aimed to evaluate whether shungite application to Umbric Podzols may affect leaf and root mitochondrial respiratory pathways, and the leaf response to temperature change. A pot culture experiment was conducted with Allium cepa L. seedlings, using soil shungite concentrations of 0, 5, 10, and 20 g kg−1 and two soil water regimes: well-watered (WW) and drying-wetting (DW) cycles. Soil water deficit increased total respiration (Vt) of onion leaves, but not roots, under low (13 °C) and high (33 °C) measurement temperature. Shungite application affected leaf Vt only at 13 °C: it increased the Vt rate under WW and decreased one under DW. An increase in the measurement temperature to 33 °C enhanced the sensitivity of leaf respiration to the inhibitor of the alternative respiratory pathway (salicylhydroxamic acid, SHAM). Shungite application increased the contribution of SHAM-sensitive pathway to the leaf Vt rate under WW, but not under the DW regime, regardless of the leaf temperature. In contrast to the SHAM-resistant pathway, the temperature sensitivity of the SHAM-sensitive rate decreased following the decrease in soil water availability. Shungite application increased the temperature sensitivity of both SHAM-sensitive and SHAM-resistant pathways under DW, and significantly decreased these parameters under WW. In summary, the decrease in temperature sensitivity of alternative SHAM-sensitive respiratory pathway with a decrease of soil water availability or shungite-related decrease of both SHAM-sensitive and SHAM-resistant leaf respiration may play an important role in enhancing the resistance of plant respiration to stress temperature.